B khoa hc v cng ngh

chng trnh KH&CN Bin phc v pht trin bn vng

kinh t-x hi, m s kc.09/06-10

Bo co tng kt khoa hc v cng ngh ti

Nghin cu sng lc cc cht c hot tnh sinh

hc theo nh hng khng sinh, gy c t bo
v chng xy ho t sinh vt bin nhm to cc
sn phm c gi tr dc dng
M s: KC.09.09/06-10
Ch nhim ti: GS.TS Chu Vn Minh
C quan ch tr ti: Vin Ho hc cc Hp cht thin nhin
Vin Khoa hc v Cng ngh Vit Nam

7388
02/6/2009

H Ni, 5 nm 2009

B khoa hc v cng ngh

chng trnh KH&CN Bin phc v pht trin bn vng
kinh t-x hi, m s kc.09/06-10

Bo co tng kt khoa hc v cng ngh ti

Nghin cu sng lc cc cht c hot tnh sinh

hc theo nh hng khng sinh, gy c t bo
v chng xy ho t sinh vt bin nhm to cc
sn phm c gi tr dc dng
M s: KC.09.09/06-10
C quan ch tr ti: Vin Ho hc cc Hp cht thin nhin
Vin Khoa hc v Cng ngh Vit Nam
Chu Vn Minh - Ch nhim ti
Phan Vn Kim - Th k ti, L Mai Hng, Phm Quc
Long, Hong Thanh Hng, Nguyn Minh H, Cng
Thung, Tng Kim Thun, Nguyn Huy Nam, Trng
Hng Lan, ng Dim Hng, on Thi Hng.

H Ni, 5 nm 2009

B khoa hc v cng ngh

chng trnh KH&CN Bin phc v pht trin bn vng
kinh t-x hi, m s kc.09/06-10

Tn ti: Nghin cu sng lc cc cht c hot tnh sinh hc theo nh

hng khng sinh, gy c t bo v chng xy ho t sinh vt bin nhm to cc
sn phm c gi tr dc dng
M s: KC.09.09/06-10
Kinh ph c cp (triu): 4,200
Thi gian thc hin: 1/2006-12/2008
Ngy

thng

nm 2009

Th trng c quan ch tr ti

Ngy

thng

nm 2009

Ch nhim ti

GS.TS Chu Vn Minh

Ngy nh gi nghim thu cp Nh nc: Ngy 19 thng 05 nm 2009

Kt lun chung: t loi Xut sc

C quan qun l ti

Ngy thng

nm 2009

b khoa hc v cng ngh

Ch tch hi ng nghim

Chng trnh kH&CN Bin kc.09/06/10

thu cp nh nc

GS.TSKH Phan tng sn

Thng tin tm tt v ti
1.Tn v m s ti:
Nghin cu sng lc cc cht c hot tnh sinh hc theo nh hng khng sinh, gy c
t bo v chng xy ho t sinh vt bin nhm to cc sn phm c gi tr dc dng
Thuc chng trnh KH&CN trng im cp nh nc KC09/06-10: Khoa hc v Cng ngh
Bin phc v pht trin bn vng kinh t x hi
M s: KC09.09/06-10
Thi gian thc hin: 1/2006-12/2008.
2. C quan ch tr ti:
C quan phi hp chnh:
3. Ch nhim ti:

Th k khoa hc:

Vin Ho hc cc Hp cht thin nhin

18 Hong Quc Vit, Cu giy, H Ni
in thoi: 04-8360830; Fax: 04-7564390
Vin Ti nguyn v Mi trng bin Hi Phng, Vin Y hc c
truyn Qun i, Vin Cng ngh Sinh hc, Vin Cng nghip
Thc phm, Cng ty Dc phm ng Dng
GS. TS. Chu Vn Minh
a ch: Vin Ho hc cc Hp cht thin nhin
Vin Khoa hc v Cng ngh Vit Nam
in thoi: 04-8363375; Fax: 04- 7564390
Email: cvminh@vast.ac.vn
TS. Phan Vn Kim
a ch: Vin Ho hc cc Hp cht Thin nhin
Vin Khoa hc v Cng ngh Vit Nam
in thoi: 04-7562378; Fax: 04- 7564390
Email: phankiem@vast.ac.vn

4. Mc tiu ca ti:
4.1. Xy dng c danh mc sinh vt bin c cht khng sinh, gy c t bo v chng
xy ho.
4.2. Xy dng c qui trnh cng ngh tch chit cc cht c hot tnh sinh hc v to
ra mt s sn phm c gi tr dc dng.
4.3. o to cn b trong lnh vc Hp cht thin nhin bin, mt lnh vc cn rt mi
m Vit nam.
4.4. Phi hp vi cc c quan lin quan a vo ng dng thc t.
5. Nhng ni dung chnh ca ti:
5.1. Thu thp mu, phn loi sinh vt bin, xc nh tn phn loi, to tiu bn, xy dng c
s d liu v ngun dc liu bin Vit Nam.
5.2. Sng lc hot tnh sinh hc cc mu sinh vt bin theo nh hng khng sinh, gy c
t bo v chng xy ho.
5.3. Nghin cu thnh phn ho hc v hot tnh sinh hc ca mt s loi sinh vt bin
c la chn thng qua qu trnh sng lc.
5.4. Nghin cu thm d kh nng khng vim t rong - to v kh nng sinh cc cht c
hot tnh sinh hc t vi sinh vt bin
5.5. Xy dng quy trnh cng ngh to ch phm, xy dng v thm nh tiu chun c s,
tin hnh cc nghin cu v dc l v ng k tiu chun sn phm v giy php lu
hnh ca B Y t cho 04 ch phm: CEFISH, BIONAMINE, HALIOTIS v HASAMIN

Danh sch nhng ngi thc hin

A. Tc gi chnh
1. GS TS Chu Vn Minh
2. TS Phan Vn Kim
3. PGS TS L Mai Hng
4. PGS TS Phm Quc Long
5. PGS TS Hong Thanh Hng
6. TS Nguyn Minh H
7. TS Cng Thung
8. PGS TS Tng Kim Thun
9. ThS Nguyn Huy Nam
10. TS Trng Hng Lan
11. TS ng Dim Hng
12. ThS on Thi Hng

Ch nhim ti
Th k ti
Vin Ho hc cc Hp cht thin nhin
Vin Ho hc cc Hp cht thin nhin
Vin Ho hc cc Hp cht thin nhin
Vin Y hc C truyn qun i
Vin Ti nguyn v Mi trng bin
Vin Cng ngh Sinh hc
Vin Cng ngh Sinh hc
Vin Cng nghip thc phm
Vin Cng ngh Sinh hc
Cng ty Dc ng Dng

B. Nhng ngi tham gia

1. TS Lu Vn Chnh
2. TS Nguyn Tin t
3. NCS Nguyn Hoi Nam
4. NCS on Lan Phng
5. NCS Trn Th Nh Hng
6. ThS Trn Hng Quang
7. ThS Trnh Thu Hng

Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN

27. BS Phm Th Dung

28. TS L Vn Ty
29. CN V Th Ninh
30. PGS TS V Mnh Hng
31. GS TS L Qu Phng
32. CN ng Trn Hon
33. BS Ng c Nhun

8. TS L Minh H

Vin HCTN

34. DS Nguyn Vn Chung

9. ThS Nguyn Xun Cng

10. CN Nguyn Phng Tho
11. CN Trn Hng Hnh
12. NCS Nguyn Hng Vn
13. ThS Nguyn Hi ng

Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN

35. TS Nguyn Vn X
36. ThS. Hong Th Minh Hin
37. ThS. Hong Th Lan Anh
38. CN Hong S Nam
39. CN Nguyn T Minh Thanh

14. ThS Hu Ngh

Vin HCTN

40. CN Hunh Quang Nng

15. NCS Nguyn Vn Thanh

Vin HCTN

16. ThS Mai Ngc Ton

17. NCS Phm Hi Yn
18. CN Phan Th Thanh Hng
19. CN Trn Anh Tun
20. NCS Trn Thu H
21. ThS. Nguyn T Kim Thy
22. Ths Li Quc Phong
23. KS Dng Vn ng
24. ThS. Trn T Minh H
25. CN Phm Linh Khoa
26. KS Nguyn Th Ln

Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin CNTP
Vin CNTP
Vin CNTP
Vin CNTP
Vin CNTP

41. PGS TS Nguyn Trng

Thng
42. ThS Nguyn Th Vn Anh
43. TS Nguyn Cng Thc
44. ThS Nguyn Vn Lnh
45. NCS. Nguyn ng Ngi
46. NCS. Trn Mnh H
47. CN Nguyn Th Hong
48. CN L Th Thy
49. NCS. Nguyn Vn Qun
50. CN V Th Lu
51. TS. m c Tin
52. ThS. Chu Th Cng

BVK H Ty
Vin CNSH
Vin CNSH
Hc vin QY
Vin KHTDTT
Vin KHTDTT
Vin KHTDTT
Cng ty
INTECPHARM
TT GD&PT Sc k
Vin CNSH
Vin CNSH
Vin CNSH
Vin CNSH
Vin NC&UD CN
Nha Trang
i hc Y HN
i hc Y HN
Vin YHCT
Vin YHCT
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin

NH GI CNG VIC SO VI HP NG
09/2006/H-TCT-KC09.09-06-10
I. DANH MC SN PHM KHOA HC V CNG NGH DNG KT QU III, IV

Stt

TN TI LIU

1
1.1
1.2
2
3
4
5
6

Bi bo, bo co khoa hc
Cng b trn tp ch quc t
Cng b trn tp ch trong nc
Sch chuyn kho
o to thc s
o to tin s
Bng c quyn sng ch
Quy trnh cng ngh chit xut v
phn lp cc cht c HT
Quy trnh cng ngh chit xut v phn
lp hot cht t hi min Holothuria
scabra
Quy trnh cng ngh chit xut v phn
lp hot cht t san h mm
Quy trnh cng ngh chit xut v phn
lp hot cht t sponge (hi min)
Quy trnh cng ngh chit xut v phn
lp hot cht t sao bin
Quy trnh cng ngh chit xut v phn
lp hot cht t sponge 2
Quy trnh cng ngh chit xut v phn
lp cht t san h mm 02
Quy trnh cng ngh chit xut v phn
lp cht t hi min Ianthella sp
Quy trnh sn xut cc thc phm
chc nng
Quy trnh sn xut vin nang Bionamine
Quy trnh sn xut vin nang Harcamin
(Haliotis)
Quy trnh sn xut thc n phng v
iu tr tim mch Cefish
Quy trnh sn xut vin nang Hasamin
C s d liu v cc loi sinh vt bin
thu thp trong ti
B h s ng k lu hnh sn phm
TP chc nng
H s ng k lu hnh sn phm
Bionamine
H s ng k lu hnh sn phm
Harcamin (Haliotis)

6.1
6.2
6.3
6.4
6.5
6.6
6.7
7
7.1
7.2
7.3
7.4
8
9
9.1
9.2

Yu cu
ca H
10
03
07
01
01
01
01
06

S lng hon
thnh
16
06
10
01
02
02
02
07

01

01

01

01

01

01

01

01

01

01

01

01

00

01

04

04

01
01

01
01

01

01

01
01

01
01

04

04

01

01

01

01

9.3
9.4

H s ng k lu hnh sn phm
Cefish
H s ng k lu hnh sn phm
Hasamin

01

01

01

01

II. DANH MC SN PHM KHOA HC V CNG NGH DNG KT QU I, II

Stt

TN SN PHM

Vin nang Bionamine t tiu

chun VSATTP
Vin Harcamin (Haliotis) t
tiu chun VSATTP
Thc n chc nng phng v
h tr iu tr bnh tim mch
Cefish t tiu chun VSATTP
Sn phm Hasamin t tiu
chun VSATTP

2
3
4

Yu cu ca
S lng
H
hon thnh
20.000 vin
20.000 vin

nh gi
chung
t ch tiu

20.000 vin

20.000 vin

t ch tiu

250 kg

350 kg

50.000 vin

50.000 vin

Vt ch tiu
t ch tiu

III. CC SN PHM KHC

- H s kt qu nghin cu thm d vi sinh vt bin v rong to bin

- H s kt qu nh gi hot tnh cc mu sinh vt bin thu thp c trong
khun kh ti
- H s cc mu dch chit v dch chit cc mu sinh vt bin thu thp c.
- K yu hi tho ti KC09.09/06-10.
Ngoi ra, ti thc hin 02 chuyn trao i khoa hc vi cc nh khoa hc
Hn Quc.

Mc lc
Danh sch nhng ngi thc hin
M u
Phn I. Tng quan, m hnh v phng php nghin cu
I. Tng quan v dc liu bin
I.1. Tng quan chung tnh hnh nghin cu trn th gii
I.2. Tng quan chung tnh hnh nghin cu trong nc
II. M hnh nghin cu v phng php nghin cu
II.1. La chn m hnh nghin cu v thit k nghin cu
II.2. Cc phng php nghin cu
Phn II. kt qu nghin cu
A. Thu thp mu v sng lc hot tnh sinh hc

Trang
1
5
7
7
7
34
37
37
40
75
75
75

Chng I. Thu thp, nh loi mu sinh vt bin v xy dng

c s d liu
I.1. Tnh hnh nghin cu nhm hi min, da gai v san h mm
75
I.2. a im v thi gian thu mu
79
I.3. Mt s nt c bn v c im mi trng ti cc a im nghin cu
80
I.4. Kt qu thu mu
84
I.5. M t cc loi sinh vt bin in hnh c kh nng cha dc liu
109
I.6. Bc u xc nh cc khu vc c kh nng khai thc ngun dc liu bin
115
I.7 Xy dng c s d liu sinh vt bin
122
I.8. Nhn xt phn thu thp mu v xy dng c s d liu
127
Chng II. X l mu, to dch chit v sng lc hot tnh 128
sinh hc
II.1. Kt qu to dch chit
128
II.2. Kt qu th hot tnh khng vi sinh vt kim nh
131
II.3. Kt qu th hot tnh c t bo
141
II.4. Kt qu nh gi hot tnh chng oxy ho ca cc mu sinh vt bin thng 151
qua phn ng bao vy gc t do (DPPH).
159
B. Nghin cu ha hc theo nh hng hot tnh sinh hc
Chng III. Nghin cu ho hc theo nh hng hot tnh
sinh hc ca mt s loi san h mm
III.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi san h mm
Sarcophyton mililatensis
III.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi san h mm
Cladiella sp.
Chng IV. Nghin cu ho hc theo nh hng hot tnh
sinh hc ca mt s loi da gai
IV.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi hi sm
Holothuria scabra
IV.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi sao bin Archaster
2

160
160
223
263
263
330

typicus
Chng V. Nghin cu ho hc theo nh hng hot tnh
sinh hc ca mt s loi hi min
V.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi san hi min
Xestospongia testudinaria
V.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi hi min Gellius
varius
V.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi hi min Ianthella
sp.
Chng VI. Tng hp cc hp cht phn lp c trong
khun kh ti
VI.1. Lp cht ditecpen dng cembranoit
VI.2. Lp cht 9,11-secosterol
VI.3. lp cht tritecpenoit saponin
VI.4. Lp cht cerebrosit v ceramit
VI.5. Lp cht axit bo khng no b brm ha
VI.6. Lp cht steroit c cu trc vng propan mch nhnh
VI.7. Lp cht steroit mang nhiu nhm hydroxyl trong phn t (polyhydroxylated
steroids)
VI.8. Lp cht glycolipit
VI.9. Cc lp cht khc
VI.10. Nhn xt
C. cc nghin cu thm d v rong, to v vi sinh vt bin

Chng VII. Nghin cu thm d kh nng khng vim t

rong, to bin vit nam
VII.1. M u
VII.2. Kt qu v tho lun
VII.3. Kt lun
Chng VIII. Nghin cu thm d kh nng sinh cc cht c
hot tnh sinh hc t vi sinh vt bin
VIII.1. M u
VIII.2. Kt qu v tho lun
VIII.3. Kt lun
D. Cc nghin cu ng dng sn xut cc sn phm t ngun
dc liu bin

371
371
405
455
485
485
489
491
494
496
497
499
504
505
510
513
513
513
514
531
533
533
534
553
559

Chng IX. NGhin cu cng ngh sn sut thc n chc 562

nng b sung omega 3 phng v h tr iu tr tim mch
IX.1. M u
562
IX.2. Kt qu v tho lun
578
IX.3. Kt lun
591
Chng X. Nghin cu qui trnh sn xut thc phm chc 592
nng tng cng th lc cho vn ng vin t Sao bin

X.1. M u
X.2. Kt qu v tho lun
X.3. Kt lun

592
605
637
Chng XI. Nghin cu bo ch v nh gi tc dng bi b 638
c th ca vin nang bo ng (Haliotis)
XI.1. M u
638
XI.2. Kt qu v tho lun
639
XI.3. Kt lun
649
Chng XII. Nghin cu cng ngh sn xut vin nang mm 650
hi sm Hasamin
XII.1. M u
650
XII.2. Kt qu v tho lun
662
XII.3. Kt lun
676
Phn III. Kt lun v kin ngh Chung
677
Danh mc cng trnh cng b
683
687
Ti liu tham kho

M u
i dng l mt ngun ti nguyn v cng ln, ni chim ti 70% din
tch b mt tri t. i dng cng l ni sinh sng ca 34 trong 36 ngnh sinh
vt trn tri t vi hn 300.000 loi ng thc vt c bit n. y chnh l
ngun cung cp v s cc sn phm t nhin qu gi t cc loi sinh vt bin nh
rong bin, rut khoang, ru bin (bryozoan), thn mm v t cc loi vi khun
bin cng nh vi khun lam. S a dng ca cc loi c th hin rt phong ph
nhng bi san h, ni m c n 1000 loi trn mt n v mt vung. Trong
, khu vc n Dng v Thi Bnh Dng c mt vng a dng sinh vt bin
nhit i ln nht trn th gii. Vit Nam nm trong khu vc bin Thi Bnh
Dng, s hu hn 1 triu km2 vng bin. Kt qu thng k n nay thng bo
c trn 12.000 loi ng thc vt bin Vit Nam, trong c rt nhiu loi c
c t hay c hot tnh sinh hc tim tng. Tuy vy, ngun ti nguyn phong ph
ny vn cha thu ht c nhiu s quan tm ca cc nh khoa hc, n nay mi
ch c mt s rt nh nhng nghin cu v lnh vc ny. Nghin cu, pht trin,
khai thc nhng ngun ti nguyn sinh vt bin hin ang l vn cp bch
khng ch nc ta m trn ton th gii.
Ngy nay, cng vi s pht trin ca khoa hc k thut, m hnh nghin cu
lin ngnh gia cc nh khoa hc thuc cc lnh vc Ho-Sinh-Y-Dc nhm tm
kim thuc t ngun hp cht thin nhin bin c p dng nhiu nc trn
th gii nh M, c, Hn Quc, Nht Bn Rt nhiu thuc mi c ngun gc
sinh vt bin c mt trn th trng do cc hng dc ln trn th gii cung
cp nh thuc iu tr ung th Ara-C (Cytarabin) c tch chit t loi Hi min
Cytotethy cryta, thuc khng sinh Phycocrythin c ngun gc t to (Red
algae)... Bn cch , hng nghin cu cc cng ngh chit xut, phn lp cc
hot cht t cc ngun dc liu bin c tr lng rt ln nh rong bin, hi sm
v cc ph thi ca ngnh cng nghip ch bin hi sn cng c quan tm c
bit. Nhng thnh qu nghin cu trong nhng nm gn y mang li li ch
kinh t v cng to ln cho nhiu quc gia trn th gii.
Vit Nam l mt nc c tim nng to ln v ti nguyn bin, vi h sinh
vt bin a dng v phong ph. Tuy nhin cho n nay, vic iu tra nghin cu
khai thc tim nng vn cn hn ch. ti khoa hc cng ngh cp Nh
nc KC09.09/06-10 Nghin cu sng lc cc cht c hot tnh sinh hc theo
nh hng khng sinh, gy c t bo v chng xy ho t sinh vt bin nhm
to cc sn phm c gi tr dc dng chng trnh KH&CN trng im cp
nh nc KC09/06-10: Khoa hc v Cng ngh Bin phc v pht trin bn
vng kinh t x hi" l mt ti khoa hc lin ngnh, c Vin Ho hc cc
Hp cht thin nhin xy dng trn c s k tha cc kt qu nghin cu ca
Vin Ho hc cc Hp cht thin nhin v Vin Ti nguyn v Mi trng bin
trong lnh vc ho hp cht thin nhin bin giai on t 1997 n nay, c bit
l tip tc nghin cu v trin khai cc kt qu t c ca ti KC 09.15
v nghin cu dc liu bin Vit Nam.

 ti c s kt hp cht ch gia Vin Ho hc cc Hp cht thin nhin

vi cc Vin Ti nguyn v Mi trng bin, Vin Y hc C truyn qun i,
Vin Cng ngh Sinh hc, Vin Cng nghip Thc phm. Mc tiu ca ti l
to c s khoa hc cho vic khai thc v s dng c hiu qu ngun ti nguyn
thin nhin bin, phc v pht trin kinh t bin v ho nhp cng ng quc t,
c th l:
1. Xy dng c danh mc sinh vt bin c cht khng sinh, gy c t bo
v chng xy ho.
2. Xy dng c qui trnh cng ngh tch chit cc cht c hot tnh sinh
hc v to ra mt s sn phm c gi tr dc dng.
3. o to cn b trong lnh vc Hp cht thin nhin bin, mt lnh vc cn
rt mi m Vit nam.
4. Phi hp vi cc c quan lin quan a vo ng dng thc t.
Trong qu trnh thc hin ti, chng ti nhn c s ch o v
gip ca Ban Ch nhim Chng trnh KC09/06-10, cng nh s gip ng
vin ca B Khoa hc v Cng ngh, Vin Khoa hc v Cng ngh Vit Nam,
cc Ban ngnh chc nng cng nhiu chuyn gia trong lnh vc ho sinh bin.
Chng ti xin chn thnh cm n s gip qu bu .

PHn I. Tng quan, m hnh v

phng php nghin cu
I. Tng quan v dc liu bin
I. 1. Tng quan chung tnh hnh nghin cu trn th gii
1. Lc s hnh thnh, pht trin v cc hng nghin cu hot cht sinh
hc bin hin nay.
Tri qua hng t nm tin ha, cc sinh vt bin t hnh thnh nn cc hp
cht ha hc v cng phc tp. Sng trong mi trng cnh tranh khc nghit,
cc sinh vt bin t sn sinh cho mnh cc hp cht c cu trc c bit
phng v trc cc mi e da ca k sn mi, ca mi trng sng v v s cc
hot ng khc nh chng s xm nhim ca cc loi h trn b mt, cnh tranh
sn mi, sinh sn. Nhng hp cht ny hot ng nh mt h thng phng v ha
hc. Chng l cht c i vi cc loi c, gy hy hoi cc m hay thm ch lm
c mi trng xung quanh chng. in hnh nh cc hp cht ditecpenoit
flexibilide v dihidroflexibilide c pht hin trong nc bin xung quanh khu
vc sinh sng ca san h mm Sinularia flexibilis. Nhng hp cht ny hot ng
nh mt h thng phng v ha hc. Chng gy c rt mnh i vi cc loi c,
hy hoi cc m nu tip xc. c rt nhiu hp cht bin c pht hin
nhng ni rt khc nghit hay t nhng sinh vt c bit, d thng v m
chng mang nhng c tnh v cng qu bu, tim tng cho vic to thuc cha
bnh cho con ngi.
Da trn c s , ngnh ha hp cht thin nhin bin v ang ngy cng
thu ht c s quan tm ca cc nh khoa hc. Sau khong 60 nm pht trin,
ngnh ha hp cht thin nhin bin t c nhng kt qu nht nh. Tuy
nhin, nhng g thu c vn cha tng xng vi kh nng thc t cng nh
mong c ca cc nh nghin cu bin trn th gii. C mt iu r rng l
nhng sinh vt sng di bin su tri qua qu trnh tin ha lu di, sut t
k s khai n nay cho nn c th khng nh rng y chnh l mi trng a
dng nht ca s sng trn tri t vi hng nghn loi sinh vt c tr, rt nhiu
trong s chng vn cn cha c khoa hc bit n. Ngi ta cho rng s a
dng v mt ha hc l chic gng phn nh s a dng v mt sinh hc ca cc
sinh vt bin [1]. Qua thi gian tm ti, nghin cu v pht trin, ngi ta phn
lp c khong 15.000 hp cht t bin. Chng thng hin din cc lp cht
steroit, tecpenoit, axt amin, alcaloit, cc hp cht phenol, hp cht thm, cc
axt bo, saponin v v s cc dng khc. Rt nhiu trong s ny th hin cc c
im dc hc c o, duy nht. Bn cnh , nhng pht hin ny cng gp
phn gii quyt cc vn lin quan n chui thc n v mi lin h vi cc
sinh vt cng sinh [2].
Ngy nay c nhiu phng php hin i v hiu qu ang c p dng vo
tm kim cc hp cht thin nhin t cc sinh vt trn t lin cng nh di y
i dng nh phng php sng lc lng ln cc hp cht theo nh hng c
hot tnh hay phng php s dng th vin cc hp cht ho hc. Vic nghin
7

cu theo phng php s dng th vin hp cht ho hc i hi phi c s tch

lu theo thi gian, nghin cu phn loi cng phu v mt cu trc v hot tnh.
Vic phn loi cng ch c gii hn trn nhng d liu dc hc bit.
Phng php sng lc theo nh hng hot tnh sinh hc th i hi phi c thit
b ng b, t tin v i ng thc hin phi c trnh v kin thc trn nhiu
lnh vc. Tuy nhin, vi thi gian nhanh, hiu qu, tin cy cao, cc phng
php sng lc ny ang c nhiu trung tm trn th gii s dng nh l mt
trong nhng cng c hng u pht hin ra cc dc t mi.
Sng lc theo nh hng hot tnh sinh hc c nh gi l mt bc tin
trong vic nghin cu, pht trin cc dc phm mi. Qua nhng qu trnh
nghin cu, ci to, vi vic p dng nhng cng ngh tin tin, ngy nay sng
lc theo nh hng hot tnh tr thnh c s chnh ca qu trnh sng lc
hng lot (HTS - high-throughput screening). Mt v d in hnh trong vic tm
kim nhng dc t mi t bin l s pht hin ra conotoxin, mt nhm hp
cht c hot tnh gim au rt hiu qu, qua phng php sng lc theo nh
hng hot tnh t loi Conus sp. Conotoxin l nhng tc nhn i khng peptit
ca cc knh ion v l th th G-protein, chng c hot tnh gim nh nhng cn
au ko di v kh nng tim tng trong iu tr mt s cn bnh khc [3]. Mt
trong cc conotoxin in hnh l Ziconotide (MVIIA) hin ang pha 3 trong
th nghim lm sng iu tr cc cn au ko di [4].
Ngy cng c nhiu ng dng t hp cht thin nhin bin c pht trin,
chng c th c s dng nh thuc tr su, trong cng ngh nano (vi cc ng
dng ca chitin, chitosan), v c bit trong y dc hc. V s hot ng dc
hc c pht hin t ngun ti nguyn v tn ny. Rt nhiu trong s cc hp
cht phn lp c th hin hot tnh chng ung th, hot tnh khng sinh rt
mnh. c khong trn di 50 hp cht hin ang c th nghim cc giai
on lm sng hoc chun b tung ra th trng c ngun gc t bin. Ngoi ra,
cc sn phm t cc dch chit, cn tinh ch s b cng c mt trn th trng
vi cc hnh thc a dng nh m phm, sn phm b dng, cc loi thc phm
thuc v mt s sn phm phc v trong y hc.
Trong s cc loi sinh vt bin c nghin cu, cc loi thuc cc ngnh Hi
min, ng vt rut khoang v Da gai cho thy kh nng sn sinh cc hp cht
c hot tnh mnh nht [5]. in hnh nh cc loi Hi min, nhng nghin cu
v ha hc ca hi min em li rt nhiu cc hp cht c gi tr. Cho n nay,
c ti 2/3 hp cht ang c th nghim lm sng hoc c mt trn th trng l
c phn lp t hi min. Cc hp cht ny th hin cc hot ng dc hc v
cng phong ph a dng nh chng ung th, khng sinh, khng vim, chng xy
ha, chng bnh mt tr nh, cha tr cc vt thng, cha au d dy, cha bnh
t min[6]. Do m nghin cu trn cc loi ny ha hn s em lai nhiu
kt qu mi, nhng dc cht mi phc v cho cc yu cu ca con ngi v ca
mi trng t nhin.

2. Cc hp cht thin nhin bin v hot tnh sinh hc t cc loi Hi

min (Sponge), Da gai (Echinoderms) v San h mm (Soft coral)
2.1. Cc hot cht sinh hc t cc loi Hi min (Sponge)
Ngnh Porifera (Hi min) bao gm rt nhiu cc ng vt c mt t thi tin
s, chng l ngun cung cp di do cc hot cht sinh hc thu ht s quan tm
c bit ca cc nh khoa hc trn th gii. c hng ngn bi bo khoa hc
lin quan n phn lp, xc nh cu trc v th nghim hot tnh sinh hc ca
cc hp cht t hi min. Trong hn 15000 hp cht phn lp t cc loi sinh vt
sng i dng, c n trn 5300 hp cht c phn lp t hi min v hng
nm, c ti hng trm hp cht tip tc c cng b, khai ph [7,8,9]. Hot
ng dc hc ca cc hp cht phn lp c t hi min c th xp vo cc
nhm chng vim, chng khi u, c ch min dch, chng vi rt, chng st rt,
khng sinh v chng h. Cc hot cht phn lp c thng tp trung vo cc
nhm cht nucleosit khng in hnh, cc tecpen, sterol, cc peptit vng, alcaloit,
axt bo v cc dn xut ca axt amin.
Mi lin h gia hi min v y hc c t thi xa khi cc thy lang thi k
La M s dng mt s loi hi min ha vi It kch thch qu trnh ng mu
hoc trn vi mt s dch chit thc vt gy m bnh nhn. T th k 18, cc
thy thuc ngi Nga v Ucraina s dng mt s loi hi min cha bnh
au lng au ngc v thp khp [6]. Cho n nm 1951 vi s pht hin ca mt
nucleosit t Cryptotethia crypta, tin cht tng hp nn Ara-C, cht chng ung
th ph bin trn th trng ngy nay, th mi quan tm ca khoa hc n cc ng
dng dc hc ca hi min mi thc s c nh thc. Ngay sau , cc cng
b v cc thnh phn axt bo, sterol v cc thnh phn khc phn lp t cc loi
ng vt bin chng minh tnh a dng ca cc hp cht bin. Nhng nghin
cu chnh l tin cho s pht trin ca cc nghin cu ha hc v hot tnh
sinh hc ca hi min hin nay. c ti hn 500 loi hi min trn th gii
c nghin cu v ha hc v hot tnh [10]. Tuy vy th s lng vn l rt
nh so vi khong 10.000 loi hi min c bit n trn th gii. Cc hp
cht t hi min lun c c nhng c tnh qu pht trin thnh thuc cha
bnh cho con ngi. Trong , cc loi thuc ba h Haliclona, Petrosia v
Discodemia u cho cc hot cht chng ung th v khng vim rt mnh, mc
d vy th nhng nghin cu v nui trng cc loi ny vn hu nh cha c
tin hnh [11].
Cc hot cht khng vim
Trong s cc hp cht phn lp c t hi min phi k n Manoalide A,
mt sestertecpen phn lp c t loi Luffatiella variabilis, hp cht ny th
hin hot tnh khng vim rt mnh trn c ch km hm PLA2 (enzim xc tc
qu trnh to arachidonic). Cho n nay, manoalide A vn c coi l cht chun
trong cc th nghim pht trin thuc khng vim theo c ch km hm PLA2
[12].
Cc nghin cu v hi min cng ch ra rng cc hp cht tecpenoit phn lp
t nhng loi ny thng cho hot tnh chng vim ng quan tm. Hi min
chnh l loi sinh vt bin mang rt nhiu cc sterol d thng. Mt trong s l
Contignasterol phn lp t Petrosia contignata, hp cht ny th hin hot tnh
khng vim rt cao. Khng nh hu ht cc loi hi min khc, contignasterol li
9

hot ng theo c ch km hm s gii phng histamine t cc bch cu. Do ,

chng c xp vo nhm cc thuc chng vim km hm c hiu bch cu
(LSAIDs) [13]. Da trn nhng c tnh m contignasterol c th
nghim trong iu tr chng hen suyn, th nghim c thc hin bi hng
InflaZyme v Aventis Pharma. Cng bi cu trc c bit phc tp v tnh bn
ng hc, hot cht ny c la chn nghin cu thay i v ti u ha cu
trc to nn mt lot cc dn xut quan trng nh IPL576,902, IPL512,602.
Cc cht ny sau c tip tc nghin cu iu tr bnh hen n giai on
lm sng pha II [14].
OH
O

H
H
H
OH

NH2

OH
H
OH

H
OH

H
OH

OH

Cogtinasterol

IPL 512,602

Cng phi k n mt s hot cht c hot tnh khng vim mnh khc nh
sestertecpen Variabilin t Icrinia variabilis, Cacospongionolide B t
Fasciospongia cavernosa v Petrosaspongiolide M t Petrospongia nigra, chng
u l nhng hot cht th hin hot tnh khng vim mnh tun theo c ch km
hm PLA2 [15]. Variabilin km hm enzim hot dch PLA2 nng IC50 = 6.9
M nhng khng cho hiu qu km hm cc enzim lin quan n qu trnh gy
vim khc nh COX-1, COX-2, 5-LOX. Hot cht ny cn th hin kh nng c
ch s mt ht neutrophil, s hnh thnh gc superoxit v s hnh thnh
leukotriene B4 (LTB4) cng nh km hm qu trnh ph n gy bi mt s tc
nhn khc nhau [16]. Theo mt vi th nghim khc, hai hp cht halipeptins A
v halipeptins B c phn lp t loi hi min Haliclona sp. Chng th hin
hot tnh khng vim mnh trn cc th nghim in vivo. ng ch l hot tnh
ca halipeptins A mnh gp 40 n 130 ln cc thuc khng vim ang s dng
trong iu tr hin nay [17].
Cc hot cht chng ung th
Bn cnh cc hot tnh khng vim th hot tnh chng khi u c bit n
nh l tnh cht hng u thng c tm thy t cc loi hi min. Hin nay
c rt nhiu hp cht c th hin hot tnh chng ung th rt mnh c xut pht
t cc nghin cu thnh phn ha hc ca hi min [18]. Cc hot cht ny c th
c phn chia vo cc nhm c c ch hot ng nh sau:
1. Cc hot cht chng qu trnh to mch.
10

2. Cc hot cht thc y qu trnh t cht ca t bo.

3. Cc hot cht c tc ng n vng i ca t bo.
4. Cc hot cht c tc ng n qu trnh tng hp cc i phn t (protein,
DNA).
5. Cc hot cht tc ng n qu trnh h hp ca ty th.
6. Cc hot cht tc ng n qu trnh nguyn phn.
7. Cc hot cht tc ng n qu trnh dn truyn tn hiu.
C rt nhiu bng chng cho thy s c mt nng cao ca protein kinase
C (PKC) lin quan n hai qu trnh bnh hc ca bnh thp khp v bnh vy
nn cng nh lin quan n cn bnh ung th. Cc hp cht t nhin bin th
hin c tnh km hm PKC rt tt. Trong s ny phi k n cc cht nh
Isoaaptamine hay debromohymenialdisine t cc loi hi min Hadromerida
v Halichondria [19, 20].
Ngoi ra cn rt nhiu hot cht chng ung th hot ng trn cc c ch v
cng phc tp khc. Mt vi trong s cc hp cht ny c nghin cu tng
i su v c ch hot ng. Agosterol A, mt hp cht tch c t loi hi
min Spongia sp. Ngi ta chng minh c rng vic lin kt ca azido
agosterol-A trn MRP1 (mt protein c hot tnh khng nhiu thuc) v lm bt
hot protein ny tng hiu qu iu tr khi u ca n [21].
Halichondrin B, hp cht c nghin cu qua nhiu giai on lm sng
khc nhau cng c nghin cu su v mt c ch tc ng. Nhng nghin
cu v mt m hc trn cc dng t bo ung th lymph v tin lit tuyn cho
thy hot cht ny c lin quan n qu trnh t cht ca t bo [22]. Vi mc
ch pht trin cc thnh t chng ung th trc kh nng khng thuc cao ca
cc dng t bo ung th hin nay, Loganzo v cc cng s tin hnh nghin
cu su trn cc dn xut tng hp t hemiasterin, mt tripeptit d thng phn
lp t Auletta sp. Nhng nghin cu ny dn n s pht trin dn xut c tn
HTI-286 chng minh vai tr ca hp cht ny trong qu trnh depolime ha cc
thoi v sc v gip ha gii s khng thuc gy bi cc P-glycoprotein i vi
thuc parataxel hay vincristine. Nhng th nghim lm sng hin ang c tin
hnh vi HTI-286 [23].
Qu trnh nghin cu tin lm sng i vi hot cht peloruside A ang rt
c quan tm, hin dc cht ny c th thu c lng ln thng qua qu trnh
tng hp v nui trng loi hi min Mycale hentscheli. Peloruside A hot ng
da trn c ch n nh cc thoi v sc thng qua qu trnh polyme ha trc tip
cc tubulin khi khng c mt cc protein lin kt vi thoi v sc. V tr hot ng
vi cc ch trn tubulin ca peloruside A tng t nh vi laulimalide nhng
khc vi v tr lin kt ca parataxel. Kt qu ny cho thy y c l s l mt
nhm hot cht chng ung th vi c ch n nh cc thoi v sc hiu qu hiu
qu hn cc nhm thuc hin nay. Hp cht ny c tip tc nghin cu pht
trin cc thuc chng ung th mi theo hng tng cng kh nng gy qu trnh
t cht ca cc t bo ung th [24].
Cc alcaloit cng ng vai tr quan trng trong hot ng chng ung th.
Trong s trn 5300 cc hp cht c phn lp t hi min, c n trn
2000 hp cht c cha nit [25]. Mt s chng hin gi ang tr thnh cc cht
u dy nhm to cc thuc chng ung th mi.
11

Mt hp cht mi hin ang c nghin cu tin lm sng trong chng ly

nhim v chng to mch l mt alcaloit c tn motuporamine, phn lp t
loi hi min Xetospongia exigua. T nhng nghin cu v cu trc v hot tnh
ca n, ngui ta tin hnh tng hp nn mt dn xut mi,
Dihydromotuporamine C, c tc dng rt cao trong ngn chn qu trnh ly lan
ca cc t bo ung th. Cc tc gi cho rng y l mt hp cht rt ng quan
tm c th c pht trin thnh cc thnh t chng ung th trong tng lai
[26].
Salicylihamide A, mt alcaloit phn lp t Haliclona sp. c cho l mt
cht km hm c hiu enzim ATPase ng thi cng th hin hot tnh c t
bo i vi cc dng t bo ung th cao gp 60 ln cc t bo bnh thng khc.
Gn y, cng c nhiu cc alcaloit mi c phn lp v th nghim hot tnh
chng khi u cho kt qu rt cao nh Naamine D mt imidazole alcaloit t
Leucetta cf. chagosensis, Jaspamide t Hemiastrella sp., hay vi N-methyl-epimanzamine D, hot cht cho kh nng dit t bo ung th rt mnh (IC50 i vi
dng B16F10 l 0.1 g/ml), ng ch hn na l hp cht Halitulin 14, mt
cu trc thuc khung bisquinolinylpyrrole mi t loi hi min Haliclona
tulearensis. Cht ny cho thy hiu qu dit t bo ung th rt mnh vi cc dng
t bo ung th bch cu chut P-388 (IC50=0.025 g/ml), t bo ung th phi
ngi A-549 (IC50=0.012 g/ml), t bo ung th rut kt HT-29 (IC50=0.012
g/ml) v t bo ung th hc sc t (IC50=0.025 g/ml) [27]. Hot cht dercitin
phn lp t loi hi min Dercitus sp. nng nM cng cho hot ng chng
ung th rt mnh i vi cc dng t bo P-388 v B16 [28].
Kirin Brewery pht trin mt dn xut ceramide mi KRN 7000 da trn
cc hp cht phn lp c t loi Agelas mauritianus to nn mt dc cht
chng ung th rt mnh. KRN 7000 th hin kh nng tng cng min dch v
kh nng chng di cn theo c ch thc y cc chc nng min dch t bo. Lm
sng pha 1 ca hp cht ny trn cc khi u rn chng t tnh khng c ca
thuc, kh nng tng cng cc yu t min dch nh inteuleukin v cc yu t
thc y hot ng i thc bo v t bo dit t nhin (NKT). Cn ch rng
mc cc t bo NKT trong mu i vi cc bnh nhn ung th v bnh nhn
mc bnh i ng thp hn so vi ngi khe mnh. Cc th nghim trn chut
chng minh rng cc t bo ung th c th b loi tr nu h thng min dch
c tng cng, c bit khi s dng KRN 7000 [29].
Cc hot cht khng vi rt
Hi min cng l ngun cung cp phong ph cc hot cht khng vi rt. Ngy
nay vic tm kim cc hot cht chng vi rt cng ang din ra khp ni trn th
gii trc cc mi e da t cc t dch bnh theo chu k do vi rt gy nn [30].
Tnh trong nm 2001, c ti 5 triu ca nhim HIV mi trn ton th gii, lm
tng tng s ngi b mc HIV-AIDS ln ti 40 triu. Cng trong nm 2001,
c 3 triu ngi cht v bnh HIV-AIDS [31]. c nhiu phng thuc c
a ra nhm i ph vi cn bnh AIDS, Nhng cc phng thuc ny ch c kh
nng lm gim s sao chp ca vi rt hay lm chm qu trnh din tin ca bnh
ch khng loi tr hon ton c vi rt ra khi c th. Nhng thuc hin nay
ch yu tp trung vo cc ch trn vi rt HIV nh nhn bit v lin kt vi v
12

ngoi protein ca vi rt, ngn chn khng cho qu trnh lin kt vi t bo v sao
m RNA xy ra, tng cng tng tc vi nhn ca t bo vt ch nhm thi hi
cc tiu phn vi rt ra khi c th. Vic tm kim cc thuc mi trong iu tr
HIV ang l yu cu cp bch. Trong nhng nm gn y c rt nhiu dc
cht c kh nng chng HIV cao c pht trin c ngun gc t sinh vt
bin. AZT, mt trong nhng thuc chng HIV u tin, xut pht t nghin
cu thay i cu trc cc nucleosit t hi min [32]. C th l hp cht Ara-A,
nucleosit phn lp t loi Cryptotethya crypta, y chnh l dn xut c ngun
gc t bin c mt trn th trng thuc ngy nay [33].
Bn cnh hot tnh khng vim, chng vi khun v dit tr su bnh, hp cht
japamide phn lp t loi hi min Thi Bnh Dng v n Dng cng cho
hot tnh khng HIV mnh nht tng c bit cc hp cht phn lp t bin.
Chng c EC50 l 0.019 M, tuy vy hot tnh ny khng cao bng hot cht AZT
(0.004 M). Mt iu ng tic l hp cht ny c tnh c t bo rt cao, do vy,
nhng nghin cu gn y ang tp trung to cc dn xut t c, tng cng hot
tnh ca jaspamide [34].
Papuamide A v B phn lp t loi hi min Theonella mirabilis v T.
swinhoei hin ang l nhng hp cht t nhin bin c trin vng nht trong vic
pht trin thnh thuc chng HIV. Th nghim sau 6 ngy nhim HIV, c hai hp
cht ny u cho hiu qu km hm qu trnh ly nhim ca HIV n cc t bo T
rt mnh v c rt thp [35]. Avarol v Avarone, hai sesquitecpen phn lp
t loi hi min Dysidea avara cng th hin hot tnh chng HIV kh cao. C
ch hot ng ca chng cng c bit n t nhiu. Hot ng ca cc hot
cht ny c cho l km hm hiu qu mt RNA vn chuyn. RNA ny ng vai
tr quan trng trong vic tng cng s sao chp ca vi rt cng nh c tham gia
vo hot ng tng hp cc enzim protease ca vi rt HIV [36]. Cng c rt
nhiu cc dn xut khc ca avarol c thng bo c hot tnh khng HIV v
chng ung th ng quan tm [37]. Nhng nghin cu trn vic nui trng loi
Dysidea avara cng thu c nhng kt qu bc u, kt qu ny chng
minh kh nng sn xut avarol trn lng ln l hon ton c th [38].
H

H
N

H
N

H
H

Papuamide A
Cng vi cc hot tnh km hm glucosidase, chng i tho ng,
callyspongym cng cho thy ph khng vi rt rng nh vi rt HIV, HBV, hay vi
rt gy bnh tiu chy Topsentin v bromotopsentin, nhng hp cht mi
phn lp t loi phn lp t loi hi min S. rueyzleri, cho hot tnh khng HSV1, VSV v vi rt dng vng A-59 [39]. Ngoi ra, mt vi hp cht chng vi rt
c phn lp t hi min nh 2-5 oligoadenylate (2-5A), Hamigeran B,
13

weinbersterol A v B, Mycalamide A [40]cng th hin hot ng mnh trn

cc dng vi rt c th nghim [41].
Cc hot cht khng sinh, khng nm.
Cc hot cht khng sinh c hot ng khng sinh thng d thu c thng
qua cc nghin cu v hp cht thin nhin. Cc hot cht thu c t hi min
cng nm trong s . Ngy nay, cc th nghim sng lc trn quy m ln i vi
cc hot tnh khng sinh, khng nm l tng i d pht trin v tr thnh
cng c hng ngy trong v s cc phng th nghim trn th gii. Tuy vy cng
cn lu rng cho n nay mi ch c mt vi thuc khng sinh trn th trng
c ngun gc t cc nghin cu trn cc hp cht thin nhin bin.
Arenosclerin A-C phn lp t loi Arenosclera brasiliensis th hin hot tnh
khng sinh rt mnh trn 12 dng vi khun th nghim cc bnh vin [6].
Cribrostatin 3 phn lp t Crebrochalina sp. cng cho hot ng khng sinh
mnh i vi dng vi sinh vt th nghim l Neisseria gonorrheae vi nng
c ch ti thiu t 0.09 g/ml, hp cht ny cng cho hot ng chng dng vi
khun khng khng sinh peniciline N. gonorrheae vi MIC t 0.39 g/ml [42].
Fascaplysin, mt hp cht c cu trc thuc nhm alcaloit 5 vng, phn lp t
loi hi min Fascaplysinopsis sp. cng c hot tnh km hm rt cao i vi cc
dng vi sinh vt th nghim nh S. aureus (0.1 g/disk), E. coli (5 g/disk), C.
albicans (1 g/disk), S. cerevisiea (1 g/disk) [43].
OH
OH
HO

OH

OH

OH

OH

OH

HOOC

H3C
HO
H2N

OH O
O

Amphotericin B
Cc nghin cu khoa hc ch ra rng rt nhiu hp cht khng nm thng
cng mang hot ng chng khi u, dit bo cao. V vy vic phn tch phn bit
cc c im chng nm v dit bo cc hp cht c hot tnh cng l iu cn
phi quan tm. Amphotericin B l mt trong nhng hp cht khng nm hiu
qu u tin c bit n. Tuy nhin vic s dng thuc ny cng cn phi lu
n cc tc dng ph in hnh nh hot ng gy c thn rt mnh. Cho n
nay th cc thuc khng nm c s dng nhiu nht l cc thuc thuc nhm
azole. Nhm cht ny hot ng ch yu theo phng php km hm s pht trin
ca nm bnh ch khng dit nm. Do m mi mt loi thuc chng nm
thuc dng azole mi ra i ngi ta phi lo ngay n vic tm kim pht hin
cc th h azole mi hiu qu hn v s khng thuc ca cc dng nm bnh gia
tng rt nhanh [44]. Trc tnh hnh , vic tm kim cc th h thuc chng
mi hiu qu hin ang c cc nh khoa hc rt quan tm.

14

Cc nghin cu tng quan v bin gn y cho thy cc hp cht phn lp

c t sinh vt bin thng cho hot ng khng nm cao vi ba dng Candida,
Aspergillus, v Cryptococcus. Vic pht hin, pht trin cc hp cht khng nm
t bin thng c thng qua cc phng php th nghim n gin hoc thng
qua chng trnh sng lc hng lot. Ngi ta cho rng cc hot cht chng nm
t sinh vt bin thng c nhng c ch hot ng c bit, c kh nng khng
nm rt cao. Cho n nay, c rt nhiu hot cht t sinh vt bin th hin hot
cht khng nm, trong , cc lp cht macrolide hay peptit thng th hin hot
tnh khng nm rt mnh, ha hn s l nhng lp cht mang li nhng hp cht
u dy to cc thuc khng nm hiu qu tng lai.
Mt lot cc hp cht d vng nh c pht hin t loi hi min thuc
chi Dysidea v Penares. Theo , hp cht (2R)-(-)-Dysidazirine cho thy hot
ng khng nm rt mnh trn hai chng nm l Candida albicans v
Saccharomyces cerevisiae (4 g/a) ng thi cng c hot tnh chng ung th
rt cao trn dng L1210 (0.27 g /ml). c bit l hp cht ny tn ti trong dch
kh ca loi hi min l rt cao, vi lng ln dn 4% trng lng th. Cc
nghin cu v tng hp ton phn ca hp cht ny cng c cng b [45].
NH2

OH
CH3

H3C
OH
HO
OH

NH2

OH

Rhizochalin
OH

HN

HN
O
Br

O
O

N
H

Jaspamide
Vo nm 1989, hot cht rhizochalin c phn lp t loi hi min
Rhizochalina incrustata bi nhm nghin cu ca Stonik [46]. Tip , mt dn
xut springolipit ca rhizochalin c tn l Oceanapiside phn lp t loi hi min
nc nng l Oceanapia phillipensis c pht hin. Hai hp cht ny th
hin hot tnh khng mnh nm Candida glataba, dng nm khng thuc
15

Fuconazole nhng khng cho hiu qu trn cc dng nm C. albicans hay l C.

krusei. V chnh s khc bit ca nhm th ng gip cho Oceanapiside c
hot ng mnh hn Rhizochalin [47]. Nh c nhc n, hp cht
japamide cng cho hot tnh khng nm rt mnh. Cc nghin cu trn c ch
hot ng ca n cho thy chng c th gy c cho cc t bo ca ng vt
c v, do vy m hp cht ny khng c quan tm pht trin na [48].
Nhm cht bengazole cng cho hiu qu khng nm rt cao. T loi hi min
Japis sp. Crews v cc cng s phn lp c 2 alcaloit bengazole A v B.
Ban u, chng cho hot ng chng giun sn Nippostrongylus braziliensis, sau
, cc hot cht ny cng chng t kh nng khng nm rt n tng trn rt
nhiu chng nm khc nhau nh: Candida albicans, Saccharomyces
carlsbergensis, C. krusei. Ngi ta cng pht hin rng chng c c ch hot
ng gn vi amphotericin B. Cc nghin cu su hn v kh nng tng hp cng
nh mi lin h gia cu trc v hot tnh ca 2 hp cht trn ang c tin
hnh nhm to ra dng thuc khng nm mi [49]. Gn y, hp cht
Kabiramide C cng c ch nhiu. Cht ny c phn lp t loi
Hexabranchus sanguineus v sau l loi Icrinia sp. Hp cht ny th hin hot
tnh km hm c hiu vi dng C. albicans nhng khng th hin hot tnh
chng khun. c bit l dn xut ca n, halichondramide A th hin hot tnh
cc mnh vi MIC t 0.2 g/ml. Tuy vy, cng ging nh nhiu loi triazole
khc, Kabiramide C v cc dn xut c kh nng gy c t bo cao v th m n
khng c pht trin tip cc giai on lm sng. Mt s macrolide chng
nm ng ch l phorboxazole A v B, mycalolide A[50].
St rt l cn bnh nguy him thng xut hin cc khu vc thuc Chu
v Chu Phi. Hu ht cc trng hp b st rt v cht l do dng k sinh trng
Plasmodium falciparum. Vic tm kim cc thuc mi c hiu, b sung, thay th
cho cc thuc chng st rt hin nay cng ang c ngy ha dc bin quan
tm [51]. Manzamine A, mt alcaloit phn lp c t rt nhiu loi hi min
khc nhau th hin nh l mt trong nhng dc t bin ha hn nht trong lnh
vc ny. Pht hin v manzamine to nn bc t ph mi trong tm kim cc
dc liu t bin. Manzamine A c tc dng chng st rt rt cao, hot ng ca
chng c cho l hot ng tng cng p ng min dch ca c th [52].
O
O
H

N
N
CH3

OCH3

OCH3

O
O
N

OH

O
6

Halichondramide

16

OCH3

OH
OH
O
OH
O

CH3

HO
OR

A: R=

O
b

B: R=

Bengazole A-B
Cc nghin cu tng hp ton phn cng nh cc nghin cu lm sng i vi
hp cht ny hin ang c gp rt thc hin rt nhiu ni trn th gii vi n
lc bin manzamine A thnh thuc khng sinh, chng st rt thc s. ng ch
l cc dn xut ca manzamine nh manzamine B, keramamine, Ircinol,
hydroxymanzamine, xestocyclaminecng th hin v s cc c tnh dc hc
th v, to nn mt lp cht rt ng quan tm ca cc nh khoa hc hin nay
[53]. Bn cnh , cc hp cht nh Di-isocyanoadociane, halichondramide
hay Kalihinol A cng chng minh c hiu qu chng st rt ca chng trn
cc th nghim in vitro [40].
O
O
H

N
OCH3
CH3

OCH3

OCH3

O
O

O
N

O
NH2
N

OH

OCH3

Kabiramide C
Mt s hot tnh khc ca cc hp cht phn lp c t hi min
Bn cnh cc hot tnh khng vim, chng khi u v khng sinh, cc hot cht
t hi min cng th hin rt nhiu hot ng dc hc a dng khc nh cc
hot tnh chng lao, chng h, iu ha c, c ch min dch hay c ch thn
17

kinhBa hp cht sterol phn lp t loi Dysidea sp. pha Bc Australia c

hot ng c ch min dch c hiu thng qua vic c ch s lin kt ca
interleukin 8, mt cytokine c kh nng thu ht cc bch cu trung tnh ti v tr
cc m thng tn. Pateamin A, mt hp cht phn lp t loi Mycale sp. c kh
nng c ch s sn sinh interleukin 2 t s hot ha cc t bo T cn li trong
pham vi hp hn [54].
Thrombin l mt enzim serin proterase c kh nng b ch cc mnh peptit t
cc fibrinogen gy nn s to thnh cc fibrin, mt thnh phn chnh ca cc cc
ng mu. Cyclothenarnide A phn lp t loi Theonella sp. gn y cho thy
s l mt lp cht km hm cc serin proterase rt hiu qu. Hp cht ny ha hn
s l mt thuc iu tr huyt khi trong tng lai. Mt hp cht khc cng rt
ng quan tm l axt Callyspongynic phn lp t Callyspongia trucata. y
l mt hp cht km hm hot ng ca -glucosidase, v m s gip duy tr
nng ng gluc thp trong mu, ha hn tr thnh mt nhn t cha tr
tiu ng hiu qu [55].
Cc hp cht t hi min cng thng c hot ng c ch thn kinh rt ng
ch . Keramidine phn lp t Agelas sp. l mt v d in hnh. y l mt tc
nhn i khng th th serotonin v ngn cn s giao tip thn kinh iu khin
bi serotonin. Mt axt amin, Dysiherbaine, t loi Dysidea herbacea cho thy
hot ng kch thch rt mnh c th gy nn ng kinh. y c coi l mt
trong nhng hp cht u dy trong pht trin thuc iu tr bnh ri lon thn
kinh [56]. Gn y, Sakai v cng s thng bo vic phn lp hp cht
neodysiherbaine A, mt axt amin c hot ng gy ng kinh mnh t loi hi
min D. herbacea. Hot ng ca hp cht ny cng tng t nh hot ng ca
dysiherbaine [57]. T loi Xestospongia sp. Gafni v cc cng s tinh ch c
hp cht xestospongin C c hot ng iu ha c rt ng ch . Hp cht ny
l mt cht c ch rt hiu qu cc th th inositol 1,4,5-triphophat v cc bm
Ca2+ cc th li t bo [58]. Bng phng php phn lp da trn sng lc hng
lot, hot cht heteronemin c ti phn lp t loi Hyrtios reticulata. Hp
cht ny th hin hot ng c ch farnesyl transferase rt ng quan tm [59].
Mi y, Yang v cc cng s phn lp c Dragmacidin D v E t loi
Dragmacidin sp., hai hp cht ny th hin hot tnh c ch enzim
serinethreonine protein phophatase. Dragmacidin D cn th hin rt tt hot tnh
c ch enzim nitric oxide synthase thn kinh (NOS), iu ny gi m cho vic
pht trin cc thuc iu tr bnh Huntington, bnh Parkinson v Alzheimer [60].
Bng 1. Tng hp mt s hp cht in hnh phn lp t hi min v dc tnh
ca chng:
Hp cht

Lp cht

Loi

Cc hp cht c hot tnh khng vim

Manoalide
Sestertecpenoit

Luffariella variabilis

Axt Dysidotronic

Sestertecpenoit

Dysidea sp.

Icrinin-1 v 2

Sestertecpenoit

Icrinia oros

18

C ch tc dng
Km hm Phospholipase
A2
Km hm Phospholipase
A2
Km hm Phospholipase
A2

Petrosaspongiolides
M-R
Spongidines A-D

Sestertecpenoit

Petrosaspongia nigra

Alcaloit

Spongia sp.

Topsentin

Alcaloit

Topsentia genitrix

Scalaradial

Sestertecpenoit

Cacospongia scalaris

Cacosponggionolide
B
Jaspaquinol
Axt Subersic
Variabilin,
IPL512,602
Congtinasterol

Sestertecpen lacton

Fasciospongia cavernosa

Ditecpen Bezenoit
Ditecpen Bezenoit
Steroit

Japis splendens
Suberea sp.
Petrosia contignata

Steroit

Petrosia contignata

Cc hp cht c hot tnh chng khi u

Isoaaptamine
Alcaloit
Debrommohymeniald Alcaloit
isin
Adociasulfat
Triterpenoid

Km hm Phospholipase
A2
Km hm Phospholipase
A2
Km hm Phospholipase
A2
Km hm Phospholipase
A2
Km hm Phospholipase
A2
Km hm Lipoxygenase
Km hm Lipoxygenase
Km hm s gii phng
histamine t bch cu
Km hm s gii phng
histamine t bch cu

Aaptos aaptos
Halichondria

c ch Protein kinase
c ch Protein kinase

Haliclona sp.

c ch Protein Motor
Knesin

Discodemolide
Laulimalide
Peloruside
Hemiasterlin, HTI286
Dictyostatin
Spongistatin 1

Tetraene lacton
Lacton vng ln
Lacton vng ln
D Tripeptit

Discodermia dissolute
Cacospogia mycofjiensis
Mycdle hentschett
Auletta sp.

Lacton vng ln
Macrolide

Corallistidae
Spongia sp.

Halichondrin B

Polyete Macrolide

Halichondria okadai

Arenastatin A

Lacton vng ln

Dysidea Arenaria

Latrunculin A
Swinholide A
Mycalolide A
Jaspamide
Neoamphimedine
Axt Elenic
Naamine D

Thiazole macrolide
Lacton vng ln
Oxazole macrolide
Alcaloit
Alkyl phenol
Alcaloit

Latrunculia magnified
Theonella swinhoei
Mycale sp.
Hemiastrella minor
Xestospongia carbonaria
Plakinastrella sp.
Leucetta cf. chagosensis

Agelasphin (KRN
7000)
Agosterol

Galactosylceramide

Agelas mauritianus

Sterol

Spongia sp.

Salicylihalamide A
macrolide
Chondropsin A v B
Crambescidins 1-4
Guanidine vng 5
Cc hp cht c hot tnh c ch min dch
Simplexide
Glycolipid

Haliclona sp.
Chondropsis sp.
Crambe crambe

Polyoxygenated
sterol
Contignasterol

Sterol

Dysidea sp.

Sterol

Petrosia contignata

Xestobergsterol A v
B

Sterol

Xestospongia berquistia

Plakortis simplex

19

polyme ha thoi v sc

c ch qu trnh polyme
ha tubulin
c ch qu trnh polyme
ha tubulin
c ch qu trnh polyme
ha tubulin
polyme ha Actin
polyme ha Actin
polyme ha Actin
c ch Topoisomerase II
c ch Topoisomerase II
c ch Topoisomerase II
c ch enzim Nitricoxide
synthetase
Hot ha t bo NKT
Bt hot s khng thuc
ca TB ung th
c ch v-ATPase
c ch v-ATPase
Cht chn knh Ca2+
c ch s sinh si ca t
bo T
c ch IL-8
c ch qu trnh gii phng
histamine
c ch qu trnh gii phng
histamine

Taurodispacamide A
Alcaloit
Agelas oroides
Pateamine A
Thiazole
Mycale sp.
Cc hp cht c hot tnh c ch thn kinh v iu ha c
Dysiherbaine
Axt amin d thng Dysidea herbacea
Keramadine

Alcaloit

Agelas sp.

Xestospongin C
Okinonenlin B
Bromotopsentin

Oxaquinolizidine
Sestertecpenoit
Alcaloit

Xestospongia sp.
Spongionella sp.
Spongosorities sp.

Penaresidin A
Alcaloit
S1319
Bezothiazole
Cc hp cht c hot tnh chng vi rt
Dragmacidin F
Alcaloit
Papuamide A - D
Peptit vng

Penares sp.
Dysidea sp.

Haplosamates A v B
Hamigeran B

Sulfamat steroit
Phenolic macrolide

Halicortex sp.
Theonella mirabilis/ T.
swinhoei
Xestospongia sp
Hamigera tarangaensis

Weinbesterol A v B

Sulfat sterol

Petrosia weinbergi

Topsentin v
Idole Alcaloit
bromotopsentin
Variolin B
Alcaloit
Avarol
Sesquiterpenoit
Hennoxazole
Bisoxazole
Cc hp cht c hot tnh chng st rt
Axisonitril-3
Sesquitecpenoit
Manzamine A
Alcaloit

c ch IL-2
c ch IL-2
Tc nhn chng th th
Glutamat
Tc nhn chng th th
serotonergic
c ch IL-3
Cha r
Tc nhn chng th th 1Adrenergic
c ch Actomyosin ATPase
Cha r
Khng virt
Khng vi rt HIV-1

Spongosorites

Khng vi rt HIV-1
Khng vi rt (herpes v
polio)
Khng vi rt (bch cu
feline, vi rt cm chut)
Khng HSV-1, VSV, A-59

Kirkpatrickia varialosa
Dysidea avara
Polyfibrospongia sp.

Khng vi rt
c ch tRNA vi rt
Khng vi rt

Acanthella klethra
Cymbastela hooperi/
Diacarnus levii
Kalihinol A
Ditecpenoit
Acanthella sp.
Cc hp cht c hot tnh chng vi sinh vt v khng nm
Discodermins B, C v Peptit vng
Discodermia kiiensis
D
Topsentiasterol sulfat Sterol
Topsentia sp.
A-E
Arenosclerin A-C
Alcaloit
Arenosclera brasiliensis
Axinellamin B-D
Alcaloit
Axinella sp.
Cribostatin
Alcaloit
Crebrochalina sp.
Acanthosterol I v J
Sulfat sterol
Acanthodendrilla sp.
Bengazole A - B
Alcaloit
Japis sp.
Oceanapiside
Glycoside
Oceanapia phillipensis
Rhizochalin
Ceramit
Rhizochalina incrustata
Spongistatin
Macrolide lactone
Hyrtios erecta
Leucascandrolide
Polyete vng ln
Leucascandra caveolata
Cc hp cht c hot tnh chng h
Kalihinene X
Isocyanotecpenoit
Acanthella cavernosa
Kalihipyran B
Isocyanotecpenoit
Acanthella cavernosa
10bIsocyanotecpenoit
Acanthella cavernosa
Formarnidokalidinol
Pseudoceratidine 2
Dn xut spermidin Pseudoceratina purpurea
Ceratinamide
Dn xut
Pseudoceratina purpurea
bromotyrosin
Formoside
Sterol diperoxit
Erylus formosus/
Lendenfeldia chondrodes
Axinyssimide
Sesquitecpen
Axinyssa sp.

20

Khng vi khun
Khng vi khun/ khng
nm (D v E)
Khng vi khun
Khng vi khun
Khng vi khun
Khng nm
Khng nm
Khng nm
Khng nm
Khng nm
Khng nm

2.2. Cc hot cht sinh hc t cc loi San h mm (Soft coral)

San h mm l nhng sinh vt thuc ngnh ng vt rut khoang
Coelenterata. Trong s , nhng loi thuc cc chi Cespitularia, Clavularia,
Gersemia, Lobophytum, Nephthea, Sarcophyton, v Sinularia c bit n
nhiu nht. Chng thng cha cc phn t mang nhiu hot tnh sinh hc. Cng
vi hi min, ng vt rut khoang chnh l loi sn sinh nhiu hp cht mi c
gi tr nht. Nhng nghin cu trc y chng minh rng khong 50% dch
chit cc loi san h mm th hin hot tnh gy c cho c. Cuc sng cng sinh
ca san h mm vi cc loi to bin to nn c im sinh hc v cng th v
ca san h mm. Rt nhiu cc hp cht th cp nh cc ditecpen dng
cembranoit t san h mm c th c to nn t nhng mi tng tc vi mi
trng sinh thi nh vy [61]. V d nh cc hp cht tecpen (thuc nhm
cembranoit) nh flexibilide v dihydroflexibilide c tm thy trong nc bin
xung quanh loi san h mm Sinularia flexibilis. Nhng hp cht ny c cho l
c hot ng nh h thng phng v ha hc ca san h mm trc cc mi e
da ca k sn mi. Chng gy c rt mnh cho c, gy lot cc m, ph hy
cc trng th tinh ca cc loi san h cng. Mt khc, mt vi tecpen cng c
pht hin t loi rong Plocamium hamatum c th gy c cho cc loi san h
mm sng quanh n nh Sinularia cruciata. Ngoi ra, cc hp cht t san h
mm cng c tc dng bo v i vi mt s loi sinh vt khc trc cc ng vt
sn mi [62]. Mt hn hp tecpen c cho l c s cho s sng cng sinh gia
loi san h mm Litophyton viridis, c kh nng tit ra cc c t ichthyotoxin,
v loi c Abudefduf leucogaster, c kh nng chng li cc c t ichthyotoxin.
Khi c s e da t bn ngoi, cc loi c ny s trn ngay vo trong m san h
mm L. viridis, do m c th trnh khi cc ng vt sn mi [63].

c rt nhiu hot tnh qu bu c pht hin t cc loi san h mm nh

hot tnh khng nm, c t bo, khng vi rt, chng to khi u, v khng vim,
[64].
S quan tm ti san h mm c bt u khi ngi ta phn lp c mt
prostaglandin [(15R)-PGA2] t loi san h sng Plexaura homomalla vo nm
1969. Tip l s th nghim i vi cc loi san h h Alcyniidea [65]. Cho
ti nay c rt nhiu cc hp cht c gi tr dc hc c pht hin t cc loi
san h mm, mt vi trong s v ang tri qua cc giai on lm sng
to cc thuc cha bnh phc v cuc sng.

21

Cc t bo ni m thng khng hot ng iu kin bnh thng. Di s

hot ha ca cc yu t to thnh mch ca t bo ung th, cc t bo ni m s
sinh si, ny n, di chuyn theo cc t bo ung th, to nn cc vi ng, thc y
s ly lan ca cc t bo ung th. S dng h thng phn ng to ng ni m in
vitro, cc nh khoa hc Hn Quc pht hin hp cht acalycixenolide E phn
lp t loi san h sng Acalycigorgia inermis. Hp cht ny cho hot ng c ch
s hnh thnh cc ng rt n tng ( nng 1 g/ml, 2.5 M). Thm vo ,
hp cht ny cng th hin hot tnh c ch s tng sinh, di chuyn v xm nhp
ca cc t bo ung th ni m cng nh c ch qu trnh to mch t bo [66].
Cc ditecpen dng cembranoit l cc hp cht c mt c thc vt v ng
vt, c bit, chng thng c pht hin t cc loi san h mm. Cc
cembranoit thng cha ng nhng hot tnh sinh hc rt tim tng, in hnh
nh cc trng hp ca sarcodictycin v eleutherobin [67].
Cc hp cht ditecpen c tn l Sarcodictyin phn lp t loi san h mm a
Trung Hi Sarcodictyon roseum vo nm 1987, hin ang l nhng ng vin nng
k cho nghin cu to thuc chng ung th. Cc hp cht ny bao gm mt
khung cu trc vng kp [8,4,0]tetradecatrien ni vi xy cng vi mt gc este
nh vo v tr C8, c th l hemiketal hoc ketal. Mt lot cc dn xut ca
sarcodictyrin c phn lp hoc tng hp thc hin cc nghin cu su
hn v mt dc hc. Nhm hp cht ny hot ng trn c ch bn ha thoi v
sc, hot ng gn ging nh hot ng ca thuc chng ung th Taxol. Nhng
nghin cu gn y chng t hot cht Sarcodictyin A, B, E cho thy hot
ng gy c t bo i vi cc t bo ung th l rt cao, c th so snh vi hot
ng ca taxol. Hot ng bn ha 90% cc tubulin ca cc hp cht ny t IC50
l t 1.7 n 6.6 M (Taxol l 4.4 M). Hn th na cc sarcodictyin cn cho ch
s khng thuc thp i vi dng t bo ung th th nghim. Nhng nghin cu
c thc hin bi cng ty Pharmacia Upjohn cho thy hot cht ny c th
c nghin cu pht trin to cc thuc chng khi u khng thuc taxol. Tuy
nhin, hot ng polyme ha tubulin ca cc sarcodictyin khng mnh bng
eleutherobin phn lp t loi san h mm Eleuthoribia sp. Hot ng ca
eleutherobin da trn vic ngng qu trnh gin phn thng qua vic bn ha cc
thi v sc. Hp cht ny cho hot ng gy c vi cc dng t bo ung th
HCT116 vi IC50 t 10.7 nM (taxol: 4.6 nM) v A2780 vi IC50 t 13.7 nM
(taxol: 6.7 nM). Nhng th nghim ca hp cht ny trn cc dng t bo ung th
khng taxol cho kt qu kh cao. Ngoi ra, eleutherobin cng th hin hot tnh
chng cc dng t bo ung th tuyn tin lit, ung th v, ung th hc sc t v
ung th bung trng. Nhng nghin cu tng hp ton phn ca cc hp cht trn
cng c thng bo. [68, 69]. Ngoi ra, cng cn rt nhiu cc cembranoit
khc phn lp t san h mm c pht hin vi hot tnh gy c t bo cao [70,
71].

22

O
O
N

O
H

N
OCH3
O

O
R2

OAc

OH

R1

OH

Eleutherobin

OH
O

Sarcodictyin A (R1=Me, R2=H)

Sarcodictyin B (R1=Et, R2=H)
Sarcodictyin C (R1=Me, R2=OH)

Cc hp cht t san h mm thng th hin hot tnh chng khi u rt tt.

Decaryiol phn lp t mt loi san h mm thuc chi Nephthea phn lp o
Fiji th hin hot tnh c t bo rt n tng (IC50 t 0.15 g/ml i vi dng t
bo MCF7). Hp cht ny c cho l hot ng da trn vic km hm c hiu
s pht trin ca t bo [72]. T mt loi san h mm Cladiella sphaeroides
Nht Bn, ngi ta pht hin ra mt hp cht mi c tn l Cladiellaperoxide.
Hot cht ny c hot ng gy c cho trng tm trong cc th nghim. Hot
ng c th t c nng ppm. Qu trnh sng lc theo nh hng hot
tnh mang li hp cht steroidal glycosit t loi san h mm C. kremfi. Hp
cht ny th hin hot tnh c t bo rt cao i vi cc dng t bo th nghim
[73]. Cng theo qu trnh sng lc theo nh hng hot tnh chng khi u, ngi
ta phn lp c hp cht sinuflexolide, dihydrosinuflexolide v sinuflexibilin
t Sinularia flexibilis. Cc hot cht c hot ng tng t cng c pht hin
cc loi thuc chi Sinularia nh cc leptocladolide A-C t loi L. leptocaldos v
S. parva [74]. Thng qua phn lp theo nh hng hot tnh sinh hc, 8 hp cht
ditectpen c khung cu trc hon ton mi bao gm cc cespitularin A-F
c phn lp t loi Cespitularia hypotenaculata. Cc hp cht ny th hin
hot ng c ch khi u rt tt i vi cc dng t bo A-594, HT-29, P388 [75].
Cc Acylsperidine phn lp t loi Sinularia sp. sng bin Okinawa l nhng
cht c tc dng mnh n dng t bo ung th A431. Cng theo mt nghin cu
c lp khc th nhng hot cht ny th hin kh nng c ch enzim H+pyrophospatase thc vt rt mnh [76]. Gn y, mt hp cht c khung 9,11secosteroid, c phn lp t loi san h mm Pachyclavularia violacea o
Togian, Indonesia. Cu trc ca hp cht ny, (24S)-3,11-dihydroxy-5,6-epoxy24-methyl-9,11-seco-5-cholestan-9-one, c xc nh qua cc nghin cu
ph. ng ch , cht ny c hot ng gy c vi cc t bo P-388, A-549,
HT-29, MEL-28 kh tt [77]. Thm vo , hai hp cht c cng khung 9,11secosteroid phn lp t loi san h cnh Pseudopterogorgia americana, c gi tr
km hm 50% s pht trin ca cc t bo ung th tuyn tin lit (LbCap), ung
th phi (Calu-3) trn h thng phn ng MTT rt n tng. Bn hp cht
secosteroid khc l isogosterones A-D phn lp t mt loi san h mm thuc
23

chi Dendronephthya th th hin kh nng c ch s sinh si, ny n ca trng c

chp Balanus amphitrite vi EC50 t 2.2 g/ml [78].
Hot ng khng vim ca hp cht OAS1000 trn cc ng vt th nghim
c chng minh. OAS1000 c tn khc l methopterosin c bn tng hp
t tin cht thuc nhm ditecpen nm vng c ng phn lp t mt loi san h
mm. OAS1000 hot ng trn c ch km hm enzim PLA2, do c hot ng
khng vim rt hiu qu. Hp cht ny hin ang c th nghim lm sng
trong iu tr thc y lm lnh vt thng [15].
Cc hp cht thuc nhm pseudopterosin l cc ditecpenoit c ng phn
lp t loi san h sng Pseudopterogorgia elisabelthae. Chng l cc hot cht
khng vim v gim au hot ng trn c ch km hm s tng hp eicosanoit
bng vic km hm c hai enzim PLA2 v v 5-LOX. Mt pht hin th v l
pseudopterosin ch km hm PMN-PLA2 m khng phi cc PLA2 t cc ngun
khc. Hot ng c hiu ny ca Pseudopterosin m ra kh nng pht trin
nhng dng thuc trn cc ch c hiu mi. Dc cht ny hin ang c sn
xut bi hng OsteoArthritis Sciences. Chng vt qua cc giai on th
nghim in vivo v ang trong cc giai on lm sng to thuc khng vim trong
tng lai. Tuy vy, dch chit san h cha pseudopterosin th c thng mi
ha trn th trng di dng cc cht ph gia trong m phm lm p da
[79,80].
R 1:

OH

OH
OH
H
OR1

OH
R 2:

OR2

OH
O
OH

Pseudopterosin A (R1=Xylose, R2=H)

Pseudopterosin E (R1=H, R2=fuctose)
Vo nm 1977, axt furanoic c cng b t loi Sinularia gonatodes
Ehrenberg, 1834. 17 nm sau , ngi ta pht hin axt ny c hot ng bt
hot enzim PLA2 nc ong. Cc dn xut tng hp sau cng th hin hot
ng hot ng c ch PLA2 rt ng ch . Hp cht ph bin nht cc loi
thuc chi Sinularia l africanene. y l mt sesquitecpen ba vng rt in
hnh. Gn y, dc tnh ca africanene cng c nghin cu. N th hin
kh nng h huyt p theo nng v hot ng khng vim trn cc dng chut
th nghim gy vim. Nhng kt qu ny l rt quan trng cho nhng nghin cu
dc hc tip theo v hp cht ny [81].

24

Cc hot ng khng sinh ca cc hp cht phn lp c t san h mm

thng khng mnh nh cc loi khc, tuy vy chng cng thu ht c s ch
ca cc phng th nghim ln trn th gii [61,71].
Hot ng khng sinh ca 4 hp cht sesquitecpen c pht hin t mt
loi san h mm thuc chi Cladiella. Bn hot cht ny u c hot ng c ch
mnh vi cc dng vi khun Streptococcus pyogenes, Staphylococcus aureus,
Enterococcus faecalis, Escherlia coli, Pseudomonas aeruginosa nng 30
g/ml [82]. Litosterol l mt steroid c 19 cacbon phn lp t loi Liptophyton
viridis. Dc cht ny th hin c ch ti thiu vi s pht trin ca vi khun
lao M. tuberculosis vi IC50 = 3.13 g/ml (>90%). Nhng nghin cu sn xut
trn quy m lng ln hp cht ny cng c thc hin nhm gii quyt vn
ngun cung cp cho cc nghin cu tip sau [83]. Cc ceramit phn lp t loi
Lobophytum crassum th hin hot tnh khng sinh tng i tt i vi cc
dng vi khun th nghim [84]. Cng t mt loi Lobophytum sp., cc
cembranoit
ditecpen nh
lobohedleolide, lobohedleolide v 17dimethylaminolobohedleolide chng t kh nng c ch s xm nhim ca
HIV-1 phn ng cp t bo (EC50 3.3-4.6 g/ml), tuy vy, cc hp cht ny
li cho hiu qu bo v t bo thp ch (50-60%) [85].
Rietone, mt sesquitecpen hydroquinole phn lp t loi Alcynonium fauri
Nam Phi cng th hin hot ng khng s sao m HIV kh cao (EC50 1.23 M
v IC50 9.32 M), mc d vy, th nghim trn cng cha xc nh c hot cht
rietone c ch s sao chp ca vi rt giai on no [86].
Bng 2. Tng hp mt s hp cht in hnh phn lp t san h mm v dc tnh
ca chng:
Hp cht

Lp cht

Cc hp cht c hot tnh khng vim

Africanene
Tcpenoit
Pseudopterosins
Ditecpenoit

Loi

C ch tc dng

Sinularia sp.
Pseudopterogorgia
elisabelthae

Km hm Phospholipase A2
Km hm Phospholipase A2 v 5LOX

Sinularia gonatodes
Sinularia flexibilis

Km hm Phospholipase A2
Tc dng trn cc th nghim ng
vt

Cc hp cht c hot tnh chng khi u

Sarcodictyin A, B, E
Cembranoit
Eleutherobin
Cembranoit
Decaryiol
Tecpenoit
Dihydroflexibilide
Ditecpenoit

Sarcodictyon roseum
Eleutheribia sp.
Nephthea sp.
Sinularia flexibilis

Bn ha thoi v sc
Bn ha thoi v sc

Sinularin

Cembranoit

Sinularia flexibilis

c ch TB KB v PS

Sinuflexolide
Cladiellaperoxide
Sinulobatin A-C

Ditecpenoit

Sinularia flexibilis
Cladiella sphaeroides
Sinularia nanolobata

c ch TB A-549, JB, HT-29, P-388

Gy c th nghim trn trng tm

Sinularia leptoclados

c ch TB KB, Hepa59T/VGH

Axt furanoic
Flexibilide

Leptocladolide

Ditecpenoit

Amphilectan
ditecpen
Norcembranoit

25

c ch TB MCF7
c ch TB Hela, BGC, 95-D, CNE,
HepG2, L-02

c ch TB L1210 v KB

Cespitularin A-F

Ditecpenoit

Cespitularia
hypotenaculata

c ch TB A-549, HT-29

Ineleganolide v
Ineleganene

Lobane
ditecpen

Sinularia inelegan

c ch TB A549, P-388

Cc hp cht c hot tnh chng vi sinh vt v khng nm

Litosterol
Steroit
Liptophyton viridis

c ch vi khun lao M. tuberculosis

Flexibilide

Ditecpenoit

Sinularia flexibilis

Ceramit
Lobohedleolide v
17dimethylaminolobohe
dleolide
Rietone

Ceramit
cembranoit
ditecpen

Lobophytum crassum
Lobophytum sp.

c ch vi khun B. subtilis, S.
aureus, S. faecalis, S. sanguis, E.
Faecalis v M. phlei
Khng sinh
Chng vi rt HIV

Sesquitecpen

Alcylnonium fauri

Chng vi rt HIV

2.3. Cc hot cht sinh hc t cc loi Da gai (Echinoderms)

Cc sinh vt thuc ngnh Da Gai (Echinoderm) bao gm nm lp: Asteroidea
(Sao Bin). Ophiuroidea (), Crinoidea (Sao Lng), Holothuroidea (Da Chut
Bin) v Echinoidea (Nhm Bin). Nhng sinh vt ny thng cha nhng thnh
phn ha hc c kh nng gy c mnh i cho c v cc vi sinh vt sng quanh
n. Nhn chung, cc hp cht c pht hin t cc sinh vt ngnh da gai thng
nm cc nhm cht ceramit c ng (Cerebroside) v cc saponin. Cng nh
cc saponin, cc hp cht ceramit c ng t cc loi Da gai thng mang cc
hot tnh gy c t bo, khng nm, khng khun Cho n nay, vai tr sinh
hc ca cc cerebroside vn cha c hiu y . Ngi ta cho rng, nhng
hp cht ny c th tham gia vo cc qu trnh truyn tn hiu mng t bo, sinh
trng v bit ha t bo, duy tr hnh thi v chc nng ca t bo V mt sinh
thi hc, cc celebroside t bin th hin kh nng chng h rt tt i vi loi
Trai xanh, mt loi thng gy rt nhiu kh khn cho tu thuyn v cc phng
tin nh c [87]. C rt nhiu cc celebroside mi c phn lp t cc loi sao
bin, nhm bin v da chut bin. Trong , ng ch c hp cht mi
reguloside A v 2 hp cht ph reguloside B c phn lp t loi sao bin
Pentaceraster regulus. Reguloside A th hin hot tnh lm lnh vt thng
kh tt. Thc nghim trn cc phn on dch chit loi sao bin Ophidiaster
ophidiamus, ngi ta tch c nm hp cht glycosphingolipid mi, bao gm
cc ophidiacerebroside A-E. Chng u th hin hot tnh gy c rt mnh i
vi dng t bo ung th L1210 [87]. Ba ganglioside, SCG1-3 phn lp t loi da
chut bin Stichopus chloronotus c hot tnh gy c i vi dng t bo PC12
theo c ch gy vim thn kinh trung ng. Mi nm, c khong 10-20 hp cht
c khung ceramit gn ng c phn lp t cc loi Da gai. Cc hp cht ny,
nh cp trn c hot tnh tng i phong ph, tuy nhin, y-dc hc
hin nay vn cha c nhiu quan tm n nhm cht ny.

26

21

O
O

HO
17

12

H
OH

19

9
30

O O
28

OH

29

NaO3SO
CH2OH

CH2OH

CH3
O

O
O

OCH3

OH

OH

OH
OH

OH

OH

Holothurin A (Xyl-Qui-Glc-Glc) v B (Xyl-Qui).

Mt nhm thnh phn khc cng rt thng c pht hin t cc sinh vt
ngnh Da Gai, l cc saponin. Cc saponin Da chut bin l cc tritecpen
glycoside, hu ht thuc khung holostane [(20S)-20-hydroxy-5-lanostane-18carboxylic acid (18-20) lactone], cn cc saponin t sao bin li l cc steroid c
ng.
Nhng tritecpen glycoside t da chut bin thng c ph hot tnh rng
nh c t bo, khng nm, tan mu, iu bin min dch c khong hn
100 hp cht thuc nhm ny c phn lp t da chut bin [88-93]. Hu
ht trong cc hp cht ny c khung cu trc holostane c bn gn vi gc ng
c nhiu nht l 6 n v ni vi cacbon v tr s 3 ca aglycon. Qu trnh nghin
cu cc hp cht thuc nhm ny c khi ng t pht hin ca cc nh khoa
hc Nht Bn vi hai hp cht chnh holothurin A v holothurin B [94,95]. Tip
sau cng c rt nhiu thng bo v cc hp cht tng t phn lp c t cc
loi da chut bin thuc lp Holothurioidea, c bit l trong khong mt thp
k gn y, c hng lot cc cng trnh nghin cu pht hin cc saponin c
b khung holostane t cc sinh vt ny. S khc bit thng thy gia cc hp
cht ny ch yu c to bi s sai khc gia cc v tr ni i trong khung, v
tr v s lng cc ng gn vi C-3 ca aglycon, cng nh cu trc ca nhnh
bn. Cc nghin cu v s khc bit trong khung cu trc v mi lin quan n
phn loi hc gia cc loi thuc chi Cucumaria c tin hnh [96,97]. Bn
cnh , nhng nghin cu v mi tng quan gia cu trc v hot tnh ca cc
tritecpen glycoside phn lp t da chut bin cng c quan tm t lu.
Thng qua cc th nghim, ngi ta nhn ra rng s sai khc v cu trc ca
phn aglycon v gc ng u lin quan n s thay i v hot tnh khng nm
ca holostane. C th l cc tritecpen gn bn phn t ng thng c hot tnh

27

cao hn cc tritecpen c gn hai ng hoc su ng. Nhng cht c cha

ng quinovose thng c hot tnh cao hn, nhng hp cht c nhnh bn dng
m thng khng c hot tnh cao bng cc hp cht c nhnh bn dng vng. T
dn n kt lun l hot tnh khng nm ca cc hp cht trn ph thuc vo
kh nng to phc ca cc phn t ng vi cc mng sterol [98]. iu ny
cng a n gi thit rng chnh s c mt nng thp cc thnh phn
saponin c tit ra t cc loi da chut bin lm suy gim s pht trin ca
trng cc loi nhm bin. Ngoi hot tnh khng nm, cc loi thuc lp
Holothutioidea cn th hin nhiu hot tnh ng ch khc. Thng qua qu
trnh tm kim yu t khng HIV c th l cc tc nhn i khng vi th th ha
ng subtype-5 (CCR5), ngi ta phn lp c hai hp cht tritecpen
glycoside. Hai hp cht ny th hin kh nng km hm CCR5 trong khi khng
c hot ng tng t i vi dng th th CXCR2 [99]. Cc intercedenside AC phn lp t loi da chut bin Mensamaria intercedens th hin hot tnh
c t bo in vitro c gi tr EC50 t 0.4-0.6 g/ml vi 10 dng t bo th nghim.
Hn na, hiu qu tiu dit ca hp cht intercedenside A i vi cc dng ung
th phi Lewis v ung th sarcom S180 chut l rt ng chu [100].
Hu ht cc nghin cu v cc saponin t sao bin c khch l bi rt
nhiu hot tnh qu bu nh dit c v bi cc hot ng dc hc nh chng
khi u, chng vi rt, tan mu, km hm s vn chuyn tn hiu thn kinh. Cc
hot tnh khng vim, gim au, v gim huyt p cng c thng bo. Mt vi
hp steroid khc li th hin hot tnh khng vi khun gram dng rt tt. Trong
vi nm tr li y, c mt s lng ln cc hp cht c tch chit t
khong 80 loi sao bin t khp ni trn th gii. Cc steroid c ng phn lp
t sao bin thng c chia thnh ba loi: asterosaponin, thnh phn c pht
hin hu ht cc loi phn tch. Cc steroid vng c ng thng thy cc
loi thuc chi Echinaster v cc polyhydroxysteroid c ng, mt lp cht cng
thng c thy nhiu loi sao bin [101]. Cc nghin cu v mi lin quan
gia cu trc v hot tnh ch ra rng, cng nh da chut bin, hot tnh ca
cc steroid c ng phn lp t sao bin ph thuc ch yu vo s lng cc tiu
phn ng v cu to ca chui bn. Tuy nhin, s nh hng ca cu trc cc
loi ng v phn aglycon n hot tnh vn cha c lm sng t. Gn y,
cc nh khoa hc Nga tin hnh nghin cu mi lin h gia cu trc ca cc
polyhydrosteroid vi hot tnh c t bo v tan mu. Tm hp cht t bn loi
sao bin khc nhau c th nghim hot ng lm tan hng cu chut v gy
c i vi s pht trin ca trng cc loi nhm bin Strongylocentrotus
intermedius. Cc tc gi pht hin ra rng, hot tnh gy c t bo ca mt s
hp cht tng t l nghch vi s lng nhm OH phn aglycon. Mt khc, c
mi lin h gia tnh c t bo v hot ng lm tan mu ca cc steroid th
nghim. Nng mui, nhit v gi tr pH l nhng yu t quan trng quyt
nh hot ng lm tan mu ca cc steroid gn ng ny. Nhng nghin cu
khoa hc cng ch ra rng nhng steroit ny cn ng vai tr l thnh phn mng
t bo cc loi sao bin, c ng gp cho vic bn ha mng kp t bo [98,101].
Gn y, mi hp cht certonardoside A-J c phn lp t loi sao bin
Certonardoa semiregularis o Komun, Hn Quc. Cu trc ton phn ca hp
cht ny c khng nh. Chng th hin ph khng vi rt rng nh HIC,
28

HSV, CoxB, EMCV v VSV, tuy vy, ch c certonardoside I v J l th hin

hot ng r rt. Mt phn t ganglioside, LMG-4, phn lp t loi Luidia
maculata cho thy hot tnh gy c i vi t bo PC12 trong s c mt ca
cc yu t sinh dng thn kinh [102]. Ngoi ra cn c mt s cc
polyhydrosteroit gn ng th hin hot tnh khng mt s dng vi khun v
hot tnh gy c t bo tng i tt.
Bng 3. Tng hp mt s hp cht in hnh phn lp t cc loi da gai v dc
tnh ca chng:
Hp cht

Lp cht

Cc hp cht c hot tnh khng vim

Reguloside A
Cerebroside

Cc hp cht c hot tnh chng khi u

Ophidiacerebrosi
Cerebroside
de A-E
SCG1-3
Ganglioside

Loi

C ch tc dng

Pentaceraster
regulus

Lm lnh vt thng

Gy c t bo ung th L210

Gy c t bo ung th

Intercedenside AC
LMG-4

Tritecpen glycoside
Ganglioside

Ophidiaster
ophidiamus
Stichopus
chloronotus
Mensamaria
intercedens
Luidia mclata

Holothurin A-B

Tritecpen glycoside

Holothuria sp.

Cc hp cht c hot tnh chng vi sinh vt v khng nm

Certonardoside I- steroidal glycoside
Certonardoa
J
semiregularis

Gy c t bo ung th PC12 theo c ch

gy vim thn kinh trung ng
Gy c 10 dng t bo ung th
Gy c t bo ung th PC12

Khng vi rt HIC, HSV, CoxB, EMCV,

VSV

3. Kh nng khai thc ngun dc liu bin, c hi v thch thc

Theo thng k, mt phn ba s thuc c mt trn th trng c ngun gc t
t nhin. C mt iu ng ngc nhin l cho n nay, hu ht cc loi thuc c
ngun gc t nhin li xut pht t trn mt t, ni ch chim 17/36 ngnh sinh
vt trn tri t. i dng, chim hn 70% din tch b mt tri t, vi ngun
a dng sinh vt v cng phong ph, trn thc t, s sn phm trn th trng t
sinh vt bin hin nay cha nhiu. S quan tm i vi vic khai thc ngun dc
liu bin mi ch c pht trin trong vng 50 nm tr li y. Cho n nay, mi
c khong 15-20 dc phm c ngun gc t bin ang trong giai on lm sng,
v hu ht l iu tr ung th, gim au hay chng vim nhim. Vy u l
nhng kh khn v ro cn i vi vic khai thc ngun dc liu bin trong thi
gian gn y? u s l trin vng trong vic khai thc s dng ngun dc liu
bin trong tng lai?
Cc sinh vt bin lun th hin nhng c im ho sinh v sinh l lin quan
ti cc qu trnh sinh hc bao gm vic sn sinh ra cc hot cht t nhin phc
v cho cc mc ch nht nh nh: bo v chng li s tn cng ca k sn mi,
s ly nhim v cnh tranh. Sng trong mi trng c bit nh vy nn hu ht
29

cc sinh vt bin u c cha nhng hp cht c cu trc c bit, duy nht. V

phng din dc hc, rt nhiu hp cht trong s ny c tnh c t bo cao, do
vy c th l gii nguyn nhn ti sao hu ht cc dc phm t bin hin nay c
hot tnh chng ung th. Cc nghin cu v ngun dc liu bin u tp trung
ch yu vo cc ngnh ng vt khng xng sng nh hi min, ng vt thn
mm, rut khoang hay da gai. Ngc li, to bin cho thy t kh nng hn trong
vic pht hin cc hp cht c hot tnh.

Cc ngnh khc

Da gai

V rut khoang

Ngnh c v

V thn mm

Ru bin

Hi min

Rong

Rong nu

Rong lc

Vi sinh vt

(AC - cc hot tnh chng ung th; AM - cc hot tnh khng sinh; AO - hot tnh chng xy ho;
IV - hot tnh trn in vivo; Other - cc hot tnh khc

Phn b hot tnh cc ngnh nm 2003

Mt vn quan trng trong vic pht trin cc ngun dc liu bin l

ngun cung cp. y c l l mt trong yu t chnh cn tr qu trnh pht trin
cc dc phm t bin. V d nh thu c 1 gam ET-743 th phi cn ti 1
tn E. turbunata. Vi halichondrin B, mt polyketid c hot tnh dit bo cao tch
chit t loi hi min Lissodendoryx sp, th t l thu hi cht so vi tng lng
mu th l rt nh: c 1 tn hi min Lissodendoryx sp th thu c 300 mg hn
hp hai halichondrin. Vi tr lng thp nh vy th vic ng dng iu tr cho
ngi gn nh l khng th. V d nu dng halichondrin iu tr cc cn bnh
ung th cho bnh nhn, ngi ta tnh ra rng trong 1 nm s tiu th khong 15 kg, v nh vy s cn n 3000-16000 tn hi min cung cp [103]. R rng l
nu ch da vo vic khai thc t t nhin pht trin thuc th gn nh l
khng th thc hin bi do s phn b t nhin ca ngun cung cp, ngoi ra vic
khai thc ny cn c th hu hoi mi trng sinh thi. Vic pht trin cc ngun
cung cp hin ang l mt kh khn trong vic khai thc ngun li ca dc liu
bin.
30

Tng hp ton phn cu trc cc hot cht t bin l mt trong cc hng i

hin nay. Tuy vy, phng php ny cng ang gp mt s kh khn nht nh v
cu trc mt s hp cht nh bryostatin hay ET-743 l rt phc tp, kh nng
tng hp kh thnh cng v hiu sut thp. PharmaMar, mt cng ty dc phm
ca Ty Ban Nha, tng hp c ET-743 da trn qu trnh bn tng hp 21
giai on mt hp cht t vi sinh vt bin Pseudomonas fluorescens. Mc d hiu
sut tng hp khng cao nhng hiu qu kinh t c th tnh n do nguyn liu
ban u c th thu c t nui cy ln men khi lng ln [104]. Ngoi ra, qu
trnh tng hp mt s hot cht ph bin khc nh halichondrin B hay bryostatin,
gambieriol, APL, KF cng c c nhng thnh cng tuy nhin hiu sut thu
c cng cha cao. S tng hp n tng nht c l thuc v MVIIA, mt peptit
c hot tnh gim au. S lng MVIIA c th t c l v hn thng qua qu
trnh tng hp [103-106].
Mt kh nng khc pht trin ngun li t nhin bin l nui trng hi
sn. S dng phng php nui trng cc sinh vt gn y thu c nhiu kt
qu tt. V d nh vic nui trng loi ru bin B. neritica thu c bryostatin
1 vi kt qu kh quan, c th phc v cho cc li ch kinh t [107].
Gn y, ngy cng c nhiu bng chng cho thy ngun gc ca rt nhiu
hp cht c hot tnh phn lp t ng vt bin khng xng sng l do cc sinh
vt ng vai tr l con mi trong chui thc n hay cc vi sinh vt sng cng sinh
(hay hi sinh) vi chng to nn. V d nh jaspalinolide, hp cht ban u c
bit n l phn lp t hi min Japis sp. Mt thi gian sau, t dng vi khun
Chondromyces crocatus ngi ta tch c mt hp cht c cu trc rt gn
vi jaspaklinolide, l chondramine D. Nghin cu v cu trc hp cht ET-743
t loi hi tiu E. turbinata, ngi ta thy safracin B, mt hp cht tch c t
loi vi sinh vt Pseudomonas fluorescens c cu trc tng t vi n.
Symplostatin 1, mt cu trc tng t vi dolastatin 10 c phn lp t
loi rong lam Symploca hydnoides. iu ny c th l gii c vi gi thit cc
loi ng vt bin tiu ho qua chui thc n hay thu nhn nhng vi sinh vt
ny t ngun nc bin. Mt s sinh vt sng cng sinh vi cc ng vt bin
cng l ngun sn sinh cc hp cht trn vt ch ca n. in hnh nh trng
hp ca bryostatin 1, nh cp cc phn trn, thc cht c l l do mt
dng sng cng sinh vi B. neritica to nn [103, 104,108,109]. T nhng kt qu
trn c th m ra mt hng mi trong vic pht trin cc dc t t bin bng
phng php nui cy ln men vi sinh vt c (cho l sn sinh ra cht ) ring
bit vi cc vt ch ca n, t c th to ra khi lng ln cc hp cht
phc v cho mc ch sn xut cng nghip. y s l mt hng i mi trong
vic nghin cu pht trin cc hp cht c gi tr phc v i sng t ngun dc
liu bin.
Ngay t nhng nm u ca ngnh ho hc hp cht t nhin bin c
nhng s quan tm ng k v hot tnh, sau , s quan tm n cc hp cht
bin c hot tnh sinh hc ngy cng tng. Trung bnh mi nm c n 10% hp
31

cht mi trong tng s hp cht bin c cng b, trong c rt nhiu nhng

hp cht th hin nhng hot tnh sinh hc qu bu. Di theo nhng nghin cu
trong thi gian qua, hi min v ng vt rut khoang lun l nhng ngnh c s
hp cht mi nhiu nht, tip theo l cc hp cht t vi sinh vt. Trong cc
hot tnh c th nghim, hot tnh chng ung th v hot tnh khng sinh lun
chim t l ln. Cc ngnh c v, da gai v hi min l nhng ngun cung cp
chnh cho nhng hot tnh ny v chng cha ng rt nhiu nhng hp cht
chng ung th, khng sinh tim tng, cha c pht hin.
Nhng nghin cu nhm tm kim pht hin cc phng thuc chng ung
th hin ang c tp trung ch yu trn cc ngnh c v, hi min, vi sinh vt
v ng vt thn mm. Bn cnh , nhng n lc tm kim thuc gim au,
khng sinh, khng vim cng ang t c nhiu thnh qu. T nhng nghin
cu , c nhng dc phm c ngun gc t bin c lu hnh trn th
trng nh Ara-A, Ara-C, Pseudopterosin... mt s khc ang bc vo nhng
giai on th nghim lm sng, ha hn s c mt trn th trng trong thi gian
gn nht (ET-743, Manzamine A, Kahalalide F...). Bn cnh nhng kt qu thu
c, ngnh ho hc hp cht thin nhin bin cng gp nhng kh khn nht
nh, v d nh ngun cung cp cc hp cht bin l c hn (nhng hp cht bin
thng chim khong 10-6% (hoc t hn) tng trng lng mu c c), vic
khai thc khi lng ln s lm hu hoi mi trng sinh thi v khng kinh t.
Phng php tng hp ho hc hay nui cy cc vi sinh vt c lin quan n qu
trnh to nn cht c l l nhng ng hng chnh trong tng lai nhm gii
quyt nhng vn trn. R rng l th gii i dng s ng mt vai tr quan
trng trong vic tm kim, pht hin v pht trin cc sn phm hu ch phc v
cho cuc sng con ngi trong tng lai khng xa.

4. Phng hng nghin cu sng lc hot tnh t ngun dc liu bin

trn th gii hin nay
Ngun hp cht t nhin lun chim c mi quan tm ln trong vic
nghin cu pht trin nhng dc phm mi, c ch cho cuc sng con ngi t
xa ti nay. S a dng v mt ho hc trong sinh gii ng mt vai tr ln
trong vic nghin cu, pht hin v pht trin nhng loi thuc mi, c gi tr
cao. Tuy nhin khai thc, sng lc cc hp cht c hot tnh mt cch c hiu
qu v khoa hc cng l mt vn ng quan tm. Ngy nay c nhiu phng
php hin i v hiu qu ang c p dng nh phng php sng lc lng
ln cc hp cht theo nh hng c hot tnh hay phng php s dng th vin
cc hp cht ho hc. Vic nghin cu theo phng php s dng th vin hp
cht ho hc i hi phi c s tch lu theo thi gian, nghin cu phn loi cng
phu v mt cu trc v hot tnh. Vic phn loi cng ch c gii hn trn
nhng d liu dc hc bit. Phng php sng lc theo nh hng hot tnh
sinh hc th i hi phi c thit b ng b, t tin v i ng thc hin phi c
trnh v kin thc trn nhiu lnh vc. Tuy nhin, vi thi gian nhanh, hiu
qu, tin cy cao, cc phng php sng lc ny ang c nhiu trung tm

32

trn th gii s dng nh l mt trong nhng cng c hng u pht hin ra

cc dc t mi.
Sng lc theo nh hng hot tnh sinh hc c nh gi l mt bc tin
trong vic nghin cu, pht trin cc dc phm mi. Qua nhng qu trnh
nghin cu, ci to, vi vic p dng nhng cng ngh tin tin, ngy nay sng
lc theo nh hng hot tnh tr thnh c s chnh ca qu trnh sng lc
hng lot (HTS - high-throughput screening) [110]. V mt s phng din nht
nh, vic phi hp cc php th khc nhau khng ch gip tng cng kh nng
sng lc cc hp cht c hot tnh m cn c th a ra nhng nhn xt quan
trng v c ch hot ng v hiu qu iu tr ca mu th (hay hot cht) .
Thm na, kt qu ca cc qu trnh sng lc ny cn c th cung cp nhng du
hiu ban u v nhng kh nng iu tr mi cho nhng mu th nghim, pht
hin nhng hot tnh cha c bit n trc [111]. Kh nng hot ng ca
cc mu c th c tin hnh trn cc th nghim tip sau nh:
Tch chit cc phn on da trn kt qu th hot tnh (bio assay-guided
fractionation).
Phn lp xc nh cu trc hot cht v tin hnh th nghim hot tnh
cht tch c (Secondary assay).
Tin hnh cc bc tng hp ho hc, tch chit quy m ln phc v cho
cc mc ch lm sng.
Mt iu cn ch trong phng php ny l kh nng phi hp hoc
tng tc ca cc thnh phn trong mu (VD nh dch chit, phn on) c th
to nn cc hiu qu tt hoc xu m khng th xc nh c t in vitro. Do ,
nhng th nghim tip sau phi c thc hin mt cc nghim tc, v phi hp
vi cc kt qu nghin cu trn in vivo.
Vo nhng nm 1970, t loi VSV Streptomyces clavuligerus ngi ta
pht hin ra axt clavulanic theo phng php sng lc theo nh hng tm kim
hot cht c ch enzim -lactamase [112]. Theo phng hng sng lc hot tnh
c ch enzim HMG-CoA reductase, ngi ta pht hin ra hp cht Mevastatin
t Penicillium citrinum [113]. Ngoi ra, mevastatin cn c bit n vi hot
tnh khng nm cao. Sau , hn hp gia axt clavulanic v amoxicilin tr
thnh hn hp khng sinh hng u, cn mevastatin c bit n nh mt thuc
chng tng lipit ng huyt rt hiu qu. Mt vi v d trn chng minh vai
tr quan trng ca phng php sng lc theo nh hng c hot tnh.

H2N-CKGKGAKCSRLMYDCCTGSCRSGKC-CONH2

Ziconotide
iu ny cng ng vi nhng sn phm t bin, ni c s a dng v cng
ln, ni n cha nhng khung cu trc mi vi hot tnh cao. Mt v d in hnh
trong vic tm kim nhng dc t mi t bin l s pht hin ra conotoxin,
mt nhm hp cht c hot tnh gim au rt hiu qu, qua phng php sng lc
theo nh hng hot tnh t loi Conus sp. Conotoxin l nhng tc nhn i
khng peptid ca cc knh ion v l th th G-protein, chng c hot tnh gim
33

nh nhng cn au ko di v kh nng tim tng trong iu tr mt s cn bnh

khc [114]. Mt trong cc conotoxin in hnh l Ziconotide (MVIIA) hin
ang pha 3 trong th nghim lm sng iu tr cc cn au ko di [115]. Theo
thng bo gn y, chng s c mt trn th trng nh mt thuc tht s vo
nm nay hoc nm sau cng vi ET-743. Nh cp phn trn, bin l kho
dc liu khng l cha c khai ph, vic tm kim nhng hp cht t bin c
hot tnh theo nh hng hot tnh sinh hc c nhiu trung tm nghin cu
s dng pht trin cc dc t mi mt cch thnh cng. Phng php ny ,
ang v s l mt trong nhng ng hng nghin cu chnh tm kim cc
hot cht mi, c ch phc v cho nhu cu pht trin nhng th h thuc mi
trong tng lai.
I.2. Tng quan chung tnh hnh nghin cu v cc cht c
hot tnh sinh hc trong nc
Theo nhng nghin cu thng k, Vit Nam c khong 12000 loi sinh vt
bin c bit n, bao gm 2038 loi c, 6000 loi ng vt y, 635 loi
rong bin v hng ngn loi ng thc vt ph du. Cc s liu thng k ca
Hooper v cng s nm 2000 cng b danh sch hn 1500 loi Hi min thuc
102 h vng bin ng. Cn nhng kt qu nghin cu hp tc nm 2002 gia
Vit Nam v Italia v bo tn da dng sinh hc bin Vit Nam pht hin 161
loi thuc 41 h thuc ngnh Hi min. Cc loi ny ch yu sng H Long,
Ct B, C T, Chn My, Hi Vn-Sn Tr. Theo bo co iu tra ca Phn vin
Hi Dng hc Hi Phng nm 2005, Vit Nam, da gai c khong 350 loi
thuc 58 h, 5 lp sng Vit Nam. Trong s , chc hn phi c nhiu loi c
cha ngun dc liu qu. Ngay t xa xa, nhn dn ta d bit s dng cc ngun
sinh vt bin vo vic cha bnh cho ngi. Theo Hi Thng Ln ng, nhiu
loi sinh vt bin c cng dng rt tt cu ngi nh Thu mu - Con sa c
v mn, khng c, tiu , cha n c tr em, cha bng. Loi sa sen tng
c ch nh cha cho ngi ta sinh kh, b b lao tn thnh ra chng huyt
tr. Trung Quc, Bo ng c xem nh c v mn, tnh bnh tc dng thanh
nhit, tr nng m , ho, bng lu, i h [116]. Cc v thuc Cn b v Hi to
c nhn dn s dng t lu cha cc chng loa lch, bu c, u v, u hch,
sng au tinh hon [117]... Nhng kinh nghim trong trong dn gian chng t cc
loi sinh vt bin s l ngun dc liu quan trng trong tng lai.
Vic nghin cu v ngun hp cht t nhin bin ca nc ta c l bt u t
nhng nm 60 -70 ca th k 20, t cho n nay cng khng c nhiu cng
trnh lin quan c cng b. Theo Lm Ngc Trm, nhng nghin cu v thnh
phn ho hc c bn trong sinh vt bin nc ta c bt u t nhng nm u
ca thp k 70. Tuy vy nhng nghin cu ny hu ht l cha y , tn mt.
u tin phi k n nhng nghin cu v ho hc cc loi rong bin ti Vin
Hi dng hc, ch yu tp trung vo cc nguyn t vi lng v protein cng nh
thnh phn c trng ca rong nh alginic, mannitol. Cc nghin cu tm hiu gi
tr dinh dng v mt s thnh phn ho hc ca mt s loi s dng ph bin
lm v lm thuc nh c mng, c i, ip s, c, c nga, hi sm...cng c
thc hin [118]. Cc nghin cu d t nhng cng ch ra kh nng tim tng

34

ca vic khai thc v s dng ngun li trn. Theo , cc ng vt khng xng

sng bin Vit Nam c cha nhiu axt bo phong ph nh cc loi san h, hi
sm, cu gai, cc phospholipid rong bin [119]. Cng theo nhng nghin cu
ca Lm Ngc Trm, cc c t bin nh tetrodomine hay tetrodotoxin cng
c xc nh 4 loi c nc (Tetryodontidae) v thy rng c lc ca chng
mnh nht gan v mu ri n trng, tinh dch, sau cng l c [118]. Cc lectin
ngng kt hng cu c pht hin 16 loi thn mm (Mollusk). Ngoi ra,
hot cht amoebocyte lysate, mt hot cht c gi tr cao trong y hc c
nghin cu kh y v kh nng khai thc v tch chit t bin c tin
hnh vo nhng nm 90 [120].
Trong cun Rong bin dc liu Vit Nam, cc tc gi c nhng nh
gi tng i h thng v ho hc v hot tnh sinh hc, cng nh tim nng khai
thc s dng ngun dc liu ny. Cun sch phn loi, nu ra nhng thnh
phn chnh ca rong bin nh cc sc t quang hp nh: chlorofil, carotenoid,
biliprotein; cc polysaccarit nh: axt alginic, agar, agarose, carrageenan, cc
glucan v ng manitol; cc nguyn t a vi lng, protein, vitamin (A, B1, B2,
B6, C, D, E, H, PP, K), v cc polyphenol, cc axt bo... in hnh nh hm
lng Vitamin C c trong rong v rong Nu c th t 500-3000 mg/kg.
Vitamin E trong rong Ascophyllum v Fucus sp c th t ti 600mg/kg kh
[121-122]. Cc axt bo dng Omega_3 (DHA) c bit n nh mt hot
cht chng xy ha cao, c nhiu gi tr dc dng nh bo v tim, iu tr x
va ng mch, chng bnh suy gim tr nhcng c thng k mt s loi
rong bin Vit Nam [123].
Trong khun kh ti nghin cu cp b v iu tra nh gi mt s hot
cht t sinh vt bin nhm to nhng ngun sn phm c gi tr cao. Vin Ho
hc cc Hp cht Thin nhin phi hp vi Vin Hi dng hc Nha Trang,
Phn Vin Hi dng hc Hi Phng v Bo tng Lch s Thin nhin Paris sng
lc hot tnh sinh hc mt s qun th loi hi min, hi sm v nghin cu su
v thnh phn ho hc mt s loi c trin vng theo cc nh hng chng ung
th v khng sinh giai on 1996-2002. Kt qu ban u cho thy kh nng to
dc liu rt kh quan. Trn 50%mu dch chit hi sm v 60% mu dch chit
hi min c hot tnh gy c cc dng t bo ung th gan ngi (HepG-2), ung
th mng tim ngi (Fl) v Ung th mng t cung (RD). Nhiu ch phm th c
hot tnh kh cao, chng hn dch chit Actinopga sp. c hot tnh chng cc
chng E. coli (MIC=50g/ml), B. subtilis v S. aureus (MIC=150 g/ml), Asp.
niger (MIC=50 g/ml) v F. oxyporum (MIC=100 g/ml). ng ch l dch
chit th ca loi hi min Agelas sp. (Aximellidae) khng ch c hot tnh c t
bo rt cao (IC50=0,2 g/ml i vi HepG-2, v 0,3 g/ml i vi Fl). m cn th
hin hot tnh khng sinh mnh vi cc chng E. coli (MIC=100 g/ml), B.
subtilis (MIC=50 g/ml) v C. albican (MIC=100 g/ml).
c bit trong giai on t 2003-2005, ln u tin mt ti khoa hc cng
ngh cp nh nc v nghin cu kh nng khai thc ngun dc liu bin Vit
Nam c thc hin bi Vin ha hc cc hp cht thin nhin, Vin Khoa
hc v Cng ngh Vit Nam. Trong hai nm, ti t chc tng cng 9 t
kho st v thu thp mu, xc nh tn phn loi, to tiu bn cho hn 405 mu
sinh vt bin, thc hin sng lc vi ba hot tnh chng xy ha, khng vi sinh
35

vt v chng ung th. Sau mt thi gian di tin hnh, qu trnh sng lc em
li mt s nh gi ban u v ngun sinh vt bin v kh nng ng dng pht
trin lm thuc. Trong s cc mu sng lc c trn 60% s sinh vt th nghim
cho hot tnh khng vi sinh vt kim nh, 25% th hin hot tnh gy c t bo
v khong 10% c hot tnh chng xy ha. Cng theo nhng kt qu nghin
cu, nhm sinh vt ngnh hi min (sponge) v da gai Vit Nam c nhiu hot
tnh qu bu nh hot tnh chng xy ho, hot tnh khng sinh v hot tnh c
t bo [124,125]. Sau l cc bc nghin cu, tch chit theo nh hng hot
tnh trn mt s loi c la chn. Trong s 9 loi c la chn nghin cu v
ho hc, c 33 hp cht c phn lp v xc nh cu trc cng nh nh gi
hot tnh sinh hc [126-129]. Cc kt qu thu c mt mt ng gp nhng
hiu bit v thnh phn cng nh hot tnh sinh hc ca cc loi sinh vt bin
Vit Nam, mt khc nhng nghin cu ny cng thnh cng trong vic pht hin
ra nhng hp cht qu bu, c tim nng pht trin thnh cc dc t trong tng
lai. in hnh nh vic phn lp v xc nh cu trc cc hp cht Holothurin A
v B t Holothuria vagabunda, l nhm hp cht c hot tnh c t bo cao v
ang thu ht c nhiu s quan tm trn th gii. Holothurin c bit n vi
nhiu hot tnh qu nh hot tnh chng vim khp dng thp v vim xng
khp [130], hot tnh khng nm v chng ung th [131]. Hp cht ny c
hng Sigma rao bn vi gi t hn kim cng: 5000 USD/1g. Ngoi ra, hai dn
xut ca Variabilin phn lp t loi Treiria sp cng cho thy hot tnh tt trn c
ba dng t bo ung th ngi cng nh hot tnh khng sinh, ha hn kh nng
ng dng trong ngnh cng nghip dc ti y.
Bn cnh , vic nghiu cu to ch phm sinh hc t ngun dc liu bin
c kh nng ng dng cao cng c tin hnh. V d nh cc ch phm
Cancialginat lm thuc cm mu ca L Th Truyn, vin nang tng lc t chit
phm Hi sm thu phn, mt s sn phm iu ch t nguyn liu v tm nh
thuc cha bng chitosan, ch phm h cholesterol mu TMC [132], ch phm h
tr iu tr ung th SALAMIN, ch phm b dng Omega-3 t sinh vt bin
bc u c nghin cu ng dng trong y hc v trong i sng. Cc nghin
cu trn thnh phn dinh dng ca mt s loi c v hi sm nh hm lng
protein, axt amin, cc khong cht, vitamin, cc hooc mn steroid nh hng
cho vic sn xut nhng sn phm thc phm dinh dng cho ngi [133-136].
Nhng kt qu ca nhng cng trnh nghin cu trn ng gp mt phn
hiu bit v ngun dc liu bin Vit Nam, qua cng gi ra nhng hng
nghin cu mi trit hn nhm khai thc ng n, c hiu qu ngun ti
nguyn v gi ny.

36

II. M hnh v phng php nghin cu

II.1. la chn M hnh nghin cu v thit k nghin cu
M hnh nghin cu c la chn trong ti l m hnh nghin cu ho
hc theo nh hng hot tnh sinh hc (Bioassay-guided chemical study).
y l m hnh nghin cu khp kn, i t nghin cu c bn n trin
khai to sn phm c th phc v cuc sng bng con ng ngn nht, nng cao
gi tr cuc sng v gp phn bo v ngun ti nguyn thin nhin. M hnh
nghin cu ny c cc nc tin tin trn th gii nh M, Php, Nht Bn,
Hn Quc ng dng thnh cng trong sng lc cc cht c hot tnh sinh hc t
Bin cng nh sinh vt trn cn. Tm tt cc bc tin hnh ca m hnh ny
c trnh by hnh 1.
Mu sinh vt bin x l
Chit bng MeOH 90%

Dch MeOH th

Th hot tnh

Khng c hot tnh

C hot tnh

Dng li hoc th
hot tnh khc

Dch chit MeOH

Tch phn on
(Chit dung mi phn cc khc nhau hoc sc k ct )

Phn on 1

Phn on 2

Th hot tnh

Th hot tnh

Phn on 3

Th hot tnh
Khng c hot tnh

Khng c hot tnh

C hot tnh
Tch cht sch v tinh
ch bng TLC/CC/HPLC

Dng li

Cht 1

Cht 2

Th hot tnh

Dng li

Cht 3

Th hot tnh

Khng c hot tnh

Dng li

C hot tnh

Cc nghin cu su hn v dc l

Th hot tnh

Khng c
hot tnh

Dng li

Hnh 1: M hnh nghin cu ho hc theo nh hng hot tnh sinh hc

37

 Thit k nghin cu
y l mt ti nghin cu lin ngnh, gii quyt c cc nhim v
t ra, chng ti bn bc v xy dng s nghin cu nh sau:

C s d
liu

X l, lu
gi mu, to
chit phm
phc v
sng lc
HTSH

Thu thp
mu sinh
vt bin.
Xc nh
tn KH v
to tiu bn

Xy dng
cc qui trnh
NCHH theo
nh hng
HTSH

Sng lc
hot tnh
Sinh hc
(Bioassay
sceening)

Cc QT
tch chit,
phn lp
cc cht c
HTSH

Cc kt qu nghin cu
trin khai trong ti KC09.15
Dy chuyn
thit b quy m
Pilot

Thc phm chc nng

HASAMIN tng cng
sinh lc t hi sm

Qui trnh sn xut nguyn liu

Thc phm chc nng

HALIOTIS t bo ng

Thc phm chc nng

CEFISH h tr iu tr
bnh tim mch t
Omega 3

Thc phm chc nng

BIONAMINE t
sao bin

Sn phm thc
thc t phc v
cuc sng

Cc th nghim dc l, c tnh bn trng din,

hiu lc ch phm (HASAMIN, BIONAMINE,
HALIOTIS v CEFISH)

Th nghim v hon thnh quy trnh sn xut

HASAMIN, BIONAMINE, HALIOTIS v CEFISH

Cc th nghim lm sng trn bnh nhn

38

ng k
sn phm

 T chc thc hin

ti KC 09.09/06-10 l ti nghin cu lin ngnh (Ho, Sinh, Y,
Dc, Cng ngh), cc mng nghin cu gn kt vi nhau v kt qu thc hin
ca mi nhm nghin cu u ng vai tr quan trng n kt qu chung ca
ti. V vy, ngay t khi xy dng cng cng nh khi bt u thc hin ti,
chng ti tp trung c lc lng cn b khoa hc v trin khai xy dng
tin nghin cu v ni dung nghin cu thng nht. Cc n v tham gia
nghin cu c phn cng nhim v r rng. Nhim v nghin cu ca tng n
v nh sau:
1) Vin Ti nguyn v Mi trng Bin Hi Phng
Chu trch nhim thu thp mu sinh vt bin, xc nh tn, phn loi, to
tiu bn mu v cc thng tin v mu. S lng mu, a im thu mu, s
chuyn kho st v cc k thut x l s b, bo qun mu theo yu cu ca
ti.
2) Vin Ho hc cc Hp cht thin nhin:
C quan ch tr ti KC09.09/06-10, chu trch nhim chung. Cc
phng nghin cu trong Vin Ho hc cc Hp cht Thin nhin tham gia vo
cc phn vic chnh bao gm:
X l mu sinh vt bin, to dch chit phc v cho sng lc hot tnh sinh
hc v nghin cu ho hc. Xy dng qui trnh x l mu thng nht.
Sng lc hot tnh sinh hc ca cc mu sinh vt bin theo cc hng sau:
Hot tnh khng vi sinh vt kim nh (Antimicrobial activity assay)
Hot tnh gy c t bo (Cytotoxic activity assay)
Hot tnh chng xi ho (Antioxidant activity assay)
Nghin cu ha hc theo nh hng hot tnh sinh hc: Tch chit v xc
nh cu trc cc hp cht c hot tnh sinh hc cao t cc i tng chn lc v
hng s dng.
- Xy dng qui trnh tch chit tng hp cc ch phm c th phc v y t
Ch phm HASAMIN t Hi sm.
Ch phm BIONAMINE t Sao bin.
Ch phm HALIOTIS t Bo ng.
Ch phm Thc n chc nng CEFISH phng nga v h tr iu tr bnh
tim mch.
Tham gia xy dng tiu chun c s cc ch phm trn, thm nh tiu
chun c s ti Cc An ton V sinh Thc phm B Y t
- Xy dng c s d liu v dc liu bin
3) Vin Y hc C truyn Qun i:
- Th c tnh cp (LD50)
-Th c tnh bn trng din
- Th hiu lc ch phm trn ng vt thc nghim
- Hon chnh h s th nghim dc l.
- To ch phm HALIOTIS t Bo ng
4) Vin Cng ngh Sinh hc:
- Thm d kh nng lm dc liu ca vi sinh vt bin, xy dng qui trnh
tch cht c hot tnh
39

- Phi hp to sn phm BIONAMINE v thng mi ho sn phm

5) Vin Cng nghip thc phm
Xy dng v tin hnh qui trnh sn xut thc phm chc nng CEFISH
phng nga v h tr iu tr tim mch
6) Cng ty Dc phm ng Dng
Sn xut th, ng k lu hnh v thng mi ho sn phm HASAMIN
7) Mt s n v khc
Cng phi hp vi Vin Ho hc cc Hp cht thin nhin trong vic thc
hin mt s cng vic lin quan n ti.
II.2. Cc phng php nghin cu
II.2.1. Phng php thu mu, x l s liu phn tch mu v xy dng c s
d liu
II.2.1.1. Phng php thu mu
- Theo quy phm thu thp mu sinh vt bin do UBKH KTNN ban hnh
nm 1981.
- Phng php thu mu sinh vt trong quan trc v phn tch mi trng
bin do B KHCN v MT ban hnh nm 2002.
- S dng phng php ln su kt hp vi cc thit b quay phim chp
nh.
C th thu mu bng nhiu dng c khc nhau: li thu mu, dao gm.
c bit cc thit b my mc phc v cho vic ln su, quay phim, chp nh rt
quan trng trong vic thu mu sinh vt bin. Di y l mt s dng c chnh
phc v cho thu thp mu vt.
- Cc thit b ln su
My nn kh, cc bnh cha kh, b qun o ln, mt n, o phi, chn
nhi, my quay camera v chp nh di nc
Ngi thu mu c trang b y cc phng tin ln su nh mt l,
bnh kh, o phi th, qun o ln, dy ln, chn nhi, ng h o su, o thi
gian, la bn .v.v. c c y cc s liu v hin trng mu vt thu, sinh
lng v tnh hnh phn b, chng ti p dng ng thi ln su vi s dng
phng php dy mt ct (transect method). Mt dy transect di 100m c t
song song vi b ti hai su khc nhau 3 v 10m hoc thay i tu theo su
thu mu. Dc mi mt ct 100m c chia lm 5 trm cc im (0 m, 25 m,
50m, 75 m v 100 m). trn mi trm thu mu t 3 im thu(mi im c din
tch khung nh lng l 1 m2
Nhn, cha:
- Nhn (Etiket) phi l loi giy bng m, khng b mc nt khi ngm trong
nc, trong cn hoc formol. Trn nhn cn th hin cc ni dung sau:
- a im thu mu:
- S ln thu mu:
- Trm thu mu:
- Cht y:
- Thi gian thu mu:
- su (khu triu):
- Din tch thu mu:
- Kinh, v :
- cha: c th cha vo hai loi - ti ni lon hoc l hoc thng n nhit, tu
theo kch c ca mu thu c m ng vo tng loi l cho ph hp.
40

II.2.1.2. Cc loi ho cht

Cht bo qun mu
+ S dng lnh bo qun mu phn tch cc cht hot tnh sinh hc.
+ Cc cht c trnh by di y l s dng vo vic bo qun v phn tch
cc mu lu tr. Thng ngi ta s dng formalin ngm mu ngoi hin
trng, t khi dng cn thay vo v tr ca formalin. Bi v s thm ca cn vo
m ng vt rt chm v vy kh nng ngn chn s phn hu ca sinh vt km.
Cn phi rt thn trng khi tip xc vi formalin, v cht ny rt c v c nguy
c gy ung th. nng thp 1ppm s gy ra s phn ng ca mt, mi, hng.
Khi lm vic vi formalin bt buc phi s dng qun o bo h, gng tay cao
su, eo khu trang v lm vic ni thong, c my ht kh. Formalin s dng
ngm ng vt c nng t 5 - 20%, tu thuc vo kch c ca c th ng vt.
Dung dch c nng t 10 - 15% thng hay c s dng nht. Nh vy vic
s dng formalin 10 - 15% ngm mu ng vt ngoi hin trng, m bo
cho mu khng b h hng. Do dung dch formalin mang tnh axit cao, thng n
mn can xi cc nhm Thn mm v Da gai.V vy bt buc khi s dng
formalin phi kim ho bng cht Borax (Na2B4O7). Cht ny c tc dng kim
ho dung dch formalin, tt nht l kim ho dung dch formalin n kim
8,2. Dung dch c kim ho ny gi l dung dch m. pha ch loi dung
dch m, c nng 10%, ngi ta pha ch 4 aux borax vo 1 gallon dung
dch formalin m c (40%). to thnh cht c nh 10%, ta pha 1 phn
cht dung dch m trn vi 9 phn nc bin. Dung dch ny s c kim ho
v to thnh dung dch ng p (isotonic) so vi m ca ng vt. Ngc li nu
pha dung dch vi nc ngt s gy ra s chnh lch p sut, c th s lm gy
mt phn c th ca ng vt.
Cht nhum mu
Trong trng hp cn thit phi nhum mu ng vt, ngi ta s dng
cht rose pegal. Pha ch dung dch ny bng cch cho 4 gam rose pegal c vo 1
lt dung dch fornmalin c.
Cht nh hnh
Nu mu trong qu trnh ngm, bo qun b co li th cn thit phi s
dng cht nh hnh lm gin c tr li v tr ban u. Cht ny c pha ch
bng cch pha 73 gam MgCl2 6H2O vo mt lt nc my.
Cht bo qun mu lu di
bo qun lu di mu (trong phng th nghim hoc cho bo tng)
thng s dng loi cn 70%. Pha ch dung dch cn 70% bt buc phi dng
nc ct, v nu dng nc cng s xy phn ng kt ta gy kh khn cho vic
phn tch mu. T l pha 737 ml cn/263 ml nc. Ring ngm mu gip xc
th cn b sung thm mt lng glicerin theo t l 70% cn + 25% nc + 5%
glicerin. trong dung dch ny phn v Kitin ca gip xc s khng b hng.

41

II.2.1.3. X l s liu v phn tch mu

- Tt c mu thu c u c chp nh trn my nh k thut s v lu gi
trong my vi tnh, su ngm trong cn 70 lu tr lm mu so snh sau
ny
- Mu dc gi n cc chuyn gia trong nc v ngoi nc gim nh theo
cc phng php chuyn dng
- S dng phn mm visual basic v Microsoft Acess, GIS xy dng c s d
liu v cc loi sinh vt cha dc liu.
- Xy dng c s d liu v cc loi sinh vt cha dc liu

II.2.2. Phng php x l mu, to dch chit, sng lc hot tnh sinh
hc
Do c nhng c trng ring, cc mu sinh vt bin i hi phi c bo
qun v x l rt cn thn trnh h hng v mt hot tnh trong qu trnh x
l. Trn c s tham kho ti liu, cng nh bn bc, trao i vi cc chuyn gia
trong nc v nc ngoi trong lnh vc ny, chng ti xy dng quy trnh x
l, to dch chit v bo qun mu phc v cho sng lc hot tnh sinh hc v xy
dng cc protocol thng nht trong nh gi hot tnh sinh hc. Cc qui trnh v
protocol ny c tun th trong sut qu trnh thc hin d n.
II.2.2.1. X l mu v to chit phm phc v sng lc hot tnh sinh hc
a. Quy trnh x l mu.
Mu sinh vt bin sau khi thu v c tin hnh bo qun theo quy trnh
thng nht sau:
- Ra sch nhiu ln bng nc ct loi mui v tp bn.
- ro nc nhit phng, p nh bng giy lc bn thm ht
nc.
- Cho tng mu vo cc ti ni lng sch, km theo k hiu mu c ghi
bng bt ch trn giy nn trnh b m hoc rch nt khi tip xc vi nc v
iu kin lnh m su.
- Lp danh sch mu sinh vt bin vi k hiu mu
Cc ti mu c lu gi trong t lnh m su (Frigor 3000 l, -20oC)
m bo mu khng b phn hu bi vi sinh vt.
b. Quy trnh to dch chit phc v sng lc hot tnh
Cc mu sinh vt bin c chit theo quy trnh sau:
Bc 1: Ly mu ra khi t lnh: Trong qu trnh ly mu phi rt thn
trng v trong t lnh m su do c s to tuyt nn cc ti mu s b dnh vo
nhau do rt d b rch khi ly mu ra. Nu ti b rch s c th lm mt k hiu
mu gy ra s c khng xc nh c mu hoc nhm ln gia cc mu vi
nhau.
Bc 2: Mu ly ra c gii ng nhit phng sau ra li
nhiu ln bng nc ct trc khi tin hnh ct nh v xay mu.
Bc 3: Mu sau khi ra c cn, c ct nh v xay trn my xay mu
(Tu theo loi mu s dng cc thit b xay v nghin mu ph hp).

42

Bc 4: Mu xay c tin hnh chit 3 ln bng MeOH trn thit b

chit siu m (Ultrasonic 2010, 950W) nhit 40-50oC, thi gian chit mi
ln ti thiu 60 pht.
Bc 5: Dch chit ca 3 ln chit c lc qua giy lc (S dng giy lc
Whatman ng knh 240nm, No 1) loi sinh khi, gp li v tin hnh ct loi
dung mi di p sut gim nhit di 50oC thu c dch c MeOH.
Bc 6:
- Dch c MeOH th c tin hnh ra loi mui qua ct c nhi
nha trao i ion Dianion HP-20 vi kch thc ct: ng knh = 30 mm v
chiu di ct l = 0,5 m.
- Dch c MeOH th c a ln ct Dianion, tin hnh ra loi mui
bng nc ct vi th tch khong 1-2 lit/mu.
- Ra ton b cht trn ct Dianion HP-20 bng MeOH cho n khi dch
ra khi ct khng cn cha cc cht ho tan ca cn dch chit ban u (Khong
2-3 lt MeOH/mu).
- Lp danh sch cc mu th loi mui.
Bc 7:
- Dch MeOH ra ct Dianion c tin hnh ct loi dung mi di p
sut gim nhit di 50oC n khi lng khng i.
- Dch chit th MeOH sau khi loi mui ly t dch tng. Dch chit
ny sau khi sy kh cho vo bnh chit c dung tch ph hp. Dch ny c ho
tan trong nc ct. Cho vo dch chit dung mi n-Hexan, bnh chit c t
trong b siu m (Ultrasonic 2010, 950W) v c siu m nhit 400C trong
10 pht. Chit phn b lin tc ba ln bng n-Hexan theo t l 1:1 v th tch.
Dch chit n-Hexan c cho vo bnh cu y trn v bc hi dung mi trong
chn khng (11mm thu ngn) bng my c quay nhit 400C. Cn thu c
cn v ho tan. Cn thu c cho vo l nh c dn nhn v nh k hiu mu
gi i th hot tnh.
- Lp nc thu c t chit phn b tip tc c chit phn b ba ln
bng CHCl3 theo t l 1:1 v th tch. Lp CHCl3 c cho vo bnh cu y trn
v bc hi dung mi trong chn khng (11mm thu ngn) bng my c quay
nhit 400C. Cn thu c cn v ho tan a vo l nh c dn nhn v
nh k hiu mu gi i th hot tnh.
- Lp nc thu c t chit phn b tip tc c chit phn b ba ln
bng Butanol theo t l 1:1 v th tch. Lp Butanol c cho vo bnh cu y
trn v bc hi dung mi trong chn khng (11mm thu ngn) bng my c quay
nhit 400C. Cn thu c cn v ho tan c l nh c dn nhn v ghi
k hiu mu, gi i th hot tnh.
- Lp nc cn li c cho vo bnh cu y trn, lm bc hi dung mi
trong chn khng (11mm thu ngn) bng my c quay nhit 400C lu tr.
V d: Mu sponge cnh xanh Gellius varius (KC-001) chit bng dung
mi MeOH c k hiu l: KC-001-M. Sau khi loi mui v tin hnh phn
on trn cc dung mi n-hexane (H), CHCl3 (C), Butanol (B), Nc (N) s c
k hiu nh sau:

43

KC-001-H: Phn on n-Hexane

KC-001-C: Phn on CHCl3
KC-001-B : Phn on Butanol
KC-001-N: Phn on nc
Bc 8: Chuyn ton b lng cn dch chit thu c vo l ng cn
dch chit c cn v dn k hiu mu (L phi m bo sch, chu c
dung mi v khng b nh hng bi iu kin lnh m su).
V d: Mu c dn nhn theo quy c nh k hiu mu nh trn
KC 09.09/06-10
KC-001-B
Ngy 27 thng 05 nm 2007
Bc 9: Cn l cha cn dch chit v tnh lng cn dch chit thu c
bng cch tr i khi lng l ban u.
Bc 10: Bo qun cc l ng cn dch chit trong t lnh m su (-20oC)
trnh b vi sinh vt phn hu.
II.2.2.2. Cc phng php sng lc hot tnh sinh hc
a) Quy trnh nh gi hot tnh khng vi sinh vt kim nh (Antimicrobial
activity assay)
Nguyn liu: Mu sinh vt bin c chun b theo mt quy trnh thng
nht xy dng trong khun kh ti KC09.15 v cc chng vi sinh vt kim
nh bao gm:
- Vi khun Gr (-): Escherichia coli (ATCC 25922 )
Pseudomonas aeruginosa (ATCC 25923 )
- Vi khun Gr (+): Bacillus subtillis (ATCC 27212 )
Staphylococcus aureus.
- Nm si: Aspergillus niger, Fusarium oxysporum.
- Nm men: Candida albicans, Saccharomyces cerevisiae.
- Khng sinh kim nh:
+ Ampicilin i vi vi khun Gr(+).
+ Tetracylin i vi vi khun Gr(-).
+ Nystatin i vi nm si v nm men.
- Cch pha khng sinh: Khng sinh pha trong dung mi DMSO100% vi
nng : Ampixilin: 50mM; Tetracylin: 10mM; Nystatin: 0.04mM.
- Mi trng nui cy vi sinh vt:
* Mi trng duy tr v bo tn ging: Saboraud Dextrose Broth (SDB)
cho nm men v nm mc. Vi khun trong mi trng Trypcase Soya Broth
(TSB).
* Mi trng th nghim: Eugon broth cho vi khun, Myco phil cho nm

Phng php tin hnh v c kt qu:

- Hot tnh khng vi sinh vt kim nh nh gi hot tnh khng sinh
ca cc mu chit c tin hnh trn cc phin vi lng 96 ging (96-well
microtiter plate) theo phng php hin i ca Vander Bergher v Vlietlinck

44

(1991) [137], hin ang c p dng ti trng i hc Dc, i hc Tng hp

Illinois, Chicago, M.
Bc1: Sng lc s b tm mu (cht) c hot tnh
- Chun b vi sinh vt:
Nm c duy tr trong mi trng dinh dng nu phn nguyn liu
v phng php. Cc chng kim nh c hot ho trc khi tin hnh th
nghim trong mi trng dinh dng dch th c bit (24 gi i vi vi khun,
48 gi i vi nm). Sau c pha long ti nng 0,5 n v Mc Fland
tin hnh th nghim.
- Chun b mu th:
- Ho tan cc chit phm trong dung dch DMSO 100% bng my Vortex
vi nng 4-10 mg/ml.
- T dung dch gc nh sang phin vi lng 96 ging, mi ging10l mu.
- Nh vo mi ging c mu sn 190l vi sinh vt hot ho.
- i chng dng:
Dy 1: Mi trng
Dy 2: Khng sinh + vi sinh vt kim nh.
- i chng m: ch c vi sinh vt kim nh.
- trong t m 370C/24 gi i vi vi khun v 300C/48 gi i vi nm.
- c kt qu:
Mu dng tnh khi nhn bng mt thng thy trong sut, khng c vi
sinh vt pht trin, ging nh hnh nh cc ging chng m tnh. Mu dng
tnh bc 1 s c tip tc th bc 2 tnh gi tr MIC (Minimmum
Inhibitory Concentration).
Bc 2: Tm nng c ch ti thiu (MIC-Minimmum Inhibitory
Concentration) ca mu (cht) c hot tnh:
Cc bc tin hnh nh bc 1: Ring cc mu c biu hin dng tnh
bc 1 c pha long theo cc thang nng thp dn, t (5- 10) thang nng
tnh gi tr c ch ti thiu m vi sinh vt b c ch pht trin gn nh
hon ton.
c kt qu:
Nng dng tnh l khng c vi sinh vt pht trin. Khi nui cy
li nng ny trn mi trng thch a kim tra, c gi tr CFU (Colony
Forming Unit) < 5 .
Mu th c MIC 200g/ml: cht sch c MIC 50g/ml l c hot
tnh.
b) Quy trnh nh gi hot tnh gy c t bo (Cytotoxic activity assay).
Nguyn liu:
- Dng t bo
Dng KB (Human epidemoid carcinoma - ung th biu m) t phng th
nghim Bioassay trng i hc Dc Illinois- USA.
Dng Fl (Fibril sarcoma of Uteus - Ung th mng t cung).
Dng Hep-G2 (Hepatocellular carcinoma- Ung th gan) t Vin VSDT
Trung ng.

45

- Mi trng nui cy t bo: DMEM (Dulbeccos Modified Eagle

Medium) hoc MEME (Minimum Essential Medium with Eagles salt). C b
sung L-glutamine, Sodium piruvat, NaHCO3, PSF (Penixillin-Streptomycin
sulfate-Fungizone); NAA (Non-Essential Amino Acids); 10% BCS (Bovine Calf
Serum).
- Tripsin-EDTA 0,05%; DMSO (Dimethyl Sulfoside); TCA(Trichloro
Acetic Acid); Tris Base; PBS (Phosphate Buffered Saline); SRB (Sulfo
Rhodamine B); Acid Acetic.
- Cc typ dng 1 ln: Bnh nui cy t bo, phin vi lng 96 ging, pipet
pasteur, cc u tip cho micropipet
- Cht chun chng dng tnh: Dng cht chun c kh nng dit t bo:
Elipticine hoc Colchicine pha trong DMSO vi nng 0.01mM.
- Mu sinh vt bin: Mu sinh vt bin c chun b theo mt quy trnh
thng nht xy dng theo khun kh ti KC09.15 v KC09.09/06-10.
- T m CO2, t lnh su- 84oC, t lnh thng, my li tm, my c Elisa;
Box Laminar PII, bnh ni t lng, cn phn tch, my o pH, bung m t bo,
knh hin vi soi ngc.
Phng php tin hnh:
- Theo phng php ca Skehan v cng s [138] v Likhiwitayawuid v
cng s [139] hin ang c p dng ti vin nghin cu ung th Quc gia ca
M (NCI) v trng i hc Dc, i hc Tng hp Illinois, Chicago, M.
- Dng t bo c gi trong Nit lng, nh thc v duy tr trong cc mi
trng dinh dng c b sung huyt thanh b ti 7-10%. Ho mu th nghim
vo dung dch DMSO 100% (4- 10mg/ml) cho bc sng lc s b.
- Pha 10 thang nng cho bc 2 tnh gi tr IC50. T bo nui cy cho
pht trin ti mc 60-70%, thay mi trng sch hot ho t bo t 18-24 gi,
lc t bo sn sng thc hin th nghim.
- T bo c x l Trpsin 0,1% cho tch khi y bnh. Ho dung dch
huyn ph t bo bng mi trng sch, ra v m s lng, pha t bo nng
3x104 t bo/ml i vi dng KB, 4x104 t bo/ml i vi dng Fl.
- Thm vo cc ging c cht chun b sn trn 190l dung dch
huyn ph t bo.
- Phin c trong t CO2 thm 3 ngy.
- Kt thc th nghim: T bo khi 3 ngy c c nh bng dung dch
TCA lnh (30-50%). Ra, kh, nhum SRB 0,4% trong axit acetic 1% v ra
li bng axit acetic 1% loi mu tha v kh, ho li bng dung dch m
Tris base 10 mM.
- c trn my ELISA bc sng 495-515nm.
Tnh kt qu:
Gi tr CS: L kh nng sng st ca t bo nng no ca cht th
tnh theo % so vi i chng. Da trn kt qu o c ca chng OD (ngy 0),
DMSO 10% v so snh vi gi tr mt quang OD (optical density) khi trn
mu tm gi tr CS(%) theo cng thc:
OD (mu) OD (ngy 0)
CS% =
x 100
OD (DMSO) OD (ngy 0)

46

Gi tr CS% sau khi tnh theo cng thc trn, c a vo tnh ton Excel
tm ra % trung bnh lch tiu chun ca php th c lp li 3 ln theo
cng thc ca Ducan nh sau: lch tiu chun
=

(xi - x )2

n-1
Cc mu c biu hin hot tnh (CS < 50%) s c chn ra th nghim
tip tm gi tr IC50.
Gi tr IC50: Dng gi tr CS ca 10 thang nng , da vo chng trnh
Table curve theo thang ga tr logarit ca ng cong pht trin t bo v nng
cht th tnh gi tr IC50.
Gi tr IC50 30g/ml i vi mu th v 4g/ ml i vi cht sch l c hot
tnh.
Cng thc: 1/y=a+blnX Trong Y: Nng cht th ; X: Gi tr CS (%)

Hnh 2 : th tnh gi tr IC50 ca mu chng

Phng php quan st trc tip s thay i cu trc mng v s lng t
bo di knh hin vi nh gi kh nng gy c t bo
Nguyn liu:
- Dng t bo
T bo ung th gan ngi (Human Hepatocellular Carcinoma Hep-G2).
T bo ung th phi (Lung cancer-Lu).
T bo ung th mng tim ngi (Rhabdosarcoma-RD). t Vin VSDT T
- Mi trng nui cy t bo: DMEM (Dulbeccos Modified Eagle
Medium) hoc MEME (Minimum Essential Medium with Eagles salt) c b
sung L- glutamine, Sodium piruvat, NaHCO3, PSF(Penixillin- Streptomycin
sulfate- Fungizone); NAA (Non-Essential Amino Acids); 10% BCS (Bovine Calf
Serum)
- Tripsin-EDTA 0,05%; DMSO (Dimethyl Sulfoside); TCA(Trichloro
Acetic acid); Tris Base; PBS (Phosphate Buffered Saline); SRB (Sulfo Rhodamine
B); Acid Acetic.
- Cc dng c dng 1 ln: Bnh nui cy t bo, phin vi lng 96 ging,
pipet pasteur, cc u tip cho micropipet
47

- Cht chun chng dng tnh:

Dng cht chun c kh nng dit t bo: Elipticine pha trong DMSO vi
nng 0.01mM
- T m CO2, t lnh su- 840C, t lnh thng, my li tm, my c Elisa;
Box Laminar PII, bnh ni t lng, cn phn tch, my o pH, bung m t bo,
knh hin vi soi ngc, knh hin vi in t.
Phng php tin hnh:
Theo phng php ca Skehan v cng s (1990) [138] v
Likhiwitayawuid v cng s (1993) [139] hin ang c p dng ti vin nghin
cu ung th Quc gia ca M (NCI) v trng i hc Dc, i hc Tng hp
Illinois, Chicago, M.
Dng t bo c gi trong Nit lng, nh thc v duy tr trong cc mi
trng dinh dng c b sung huyt thanh b ti 7-10%. Cc mu c chn lc
c kt qu gy c t bo dng tnh. Ho mu th nghim vo dung dch
DMSO 100% (4-10mg/ml) v sau pha di nng nh trong th nghim hot
tnh gy c t bo
T bo nui cy cho pht trin ti mc 60-70%, thay mi trng sch
hot ho t bo t 18-24 gi, lc t bo sn sng thc hin th
nghim.
T bo c x l Trpsin 0,1% cho tch khi y bnh. Ho dung dch
huyn ph t bo bng mi trng sch, ra v m s lng, pha t bo nng
3-4x104 t bo/ml.
Thm vo cc ging c cht chun b sn trn 190 l dung dch huyn
ph t bo.
Phin c trong t CO2
Quan st s thay i cu trc mng t bo di knh hin vi soi ngc theo
thi gian v chp nh sau 24h; 48h v 72h.
Cc hnh nh thay i cu trc r nt s c quan st di knh hin vi chp nh
c) Quy trnh th hot tnh chng xy ho ca cc mu sinh vt bin thng qua
phn ng bao vy gc t do DPPH (Antioxidant activity assay- DPPH free radical
scavenging).
Nguyn liu, my mc th nghim:
1,1-diphenyl-2-picrylhydrazyl (Sigma, USA).
Axit ascorbic (Sigma, USA); Ethanol 96% (Vit Nam).
Dimethylsulphoxid (DMSO) 100% (Sigma, USA).
Phin vi lng 96 ging (Conning - M).
Bn mng Silicagel RP-18 F254s (Art. 1.05559).
Pippetman (Gilson - Php).
My c Elisa.
Mu sinh vt bin
Mu sinh vt bin c chun b theo mt quy trnh thng nht xy
dng trong khun kh ti KC 09.09/06-10.
Phng php tin hnh:

48

+ Phng php sng lc s b kh nng by gc t do trn phin 96

ging:
Th nghim chng xy ho da trn nguyn l: DPPH c kh nng to ra
cc gc t do bn trong dung dch EtOH bo ho. Khi cho cc cht th nghim
vo hn hp ny, nu cht c kh nng lm trung ho hoc bao vy cc gc t do
s lm gim cng hp ph nh sng ca cc gc t do DPPH. Hot tnh
chng xy ho c nh gi thng qua gi tr hp ph nh sng ca dch th
nghim so vi i chng khi c trn my Elisa bc sng 515nm .
Chun b mu th nghim:
- To dung dch c cc gc t do: DPPH 300M trong ethanol.
- Mu sinh vt bin pha trong DMSO 100% vi nng 4mg/ml.
- Chng dng tnh 1 mM Flavonoid hoc 5mM axit Acscorbic.
Tin hnh th nghim:
Trn mu v DPPH trn phin 96 ging.
Dy th nghim: 10l mu +190l DPPH trong Ethanol.
Dy chng m tnh: 10l DMSO 10% +190l DPPH trong ethanol.
Dy Blank (mu trng): 10l mu +190l Ethanol.
Dy chng dng tnh: 10l mu +190l DPPH trong ethanol.
Phin c bc kn trnh nh sng mi trng v trong t m 370C
trong 2 gi. c kt qu trn my Elisa bc sng 515nm [140].
Tnh kt qu:
Tnh gi tr % hot ng SC% (Scavenging capacity):
Gi tr trung bnh ca SC%. c a vo chng trnh x l s liu
Excell Window tm ra % trung bnh lch tiu chun ca php th c lp
li 3 ln theo cng thc:
OD th nghim - OD mu trng
SC% = [100 x 100]
OD chng m tnh
lch tiu chun tnh theo cng thc ca Ducan nh sau:

(xi - x )2

n-1
Gi tr hot ng SC% >50% mu c coi l c biu hin hot tnh s
c chn ra tm gi tr IC50.
Tm gi tr c ch IC50
Pha mu theo 5 thang nng , gi tr IC50 c a vo chng trnh Table
Curve, thng qua nng cht th v % hot ng ca cht th tnh ra nng
ca cht th nghim m 50% cc gc t do c to bi DPPH c trung
ho bi cht th theo cng thc:
1/y=a+blnX
Trong Y: Nng cht th X: Gi tr SC (%)
+ Phng php th nghim trc tip trn bn mng

49

Phng php hin sinh hc trc tip trn bn mng chy sc k (TLC
Bioautographic assays) l phng php n gin v r tin nht, d dng thc
hin v i hi rt t cc trang thit b pht hin cc thnh phn cht c hot
tnh. Phng php ny ng vai tr quan trng trong vic nh v v xc nh c
tnh ca cht trong qu trnh phn lp cht theo nh hng hot tnh sinh hc, l
s kt hp vic chy sc k bn mng vi th nghim sinh hc ti ch. N cho
php nh v v tr cht c hot tnh trn bn mng trong mt hn hp cht nn.
Tuy nhin, phng php ny ch c tc dng ch yu l pht hin ra cc cht,
thng l cc cht dn ng cho cc nghin cu tip theo.
Trong phn th nghim sng lc hot tnh chng xy ho thng qua th
nghim bt gi cc gc t do tip theo, chng ti p dng phng php phun
sng gc 2,2-diphenyl-1-picrylhydrazyl (DPPH) ln bn mng chy cht th
(phng php TLC-DPPH).
Mu c chm trn bn mng vi b rng 9-10mm v phn tch trn bn
nhm RP-18 (10x20cm). V tr khi u l 12mm t c hai mt v 10mm t y
phin. Phin c pht trin trn h dung mi thch hp. Sau khi kh 15 pht,
phin c nhng hoc phun sng vi dung mi DPPH 0,04% trong 5 giy. Sau
khi nhng 2 pht, nh c lu gi di nh sng kh kin. Hot tnh by gc t
do c xc nh bng chng trnh phn mm qut nh, thng qua s bin thin
ca cng mu ca cht th trn cc vng khc nhau ca phin. Di cht c
mu vng sng trn nn mu ta chnh l vng c hot tnh bao vy cc gc t do
lm thay i mu ca phn ng.
d. nh gi hot tnh chng long xng thng qua cc c ch kch thch s bit
ha ca nguyn bo xng MC3T3-E1.
Cc th nghim hot tnh chng long xng c tin hnh ti Trng
i hc Dc, i hc Quc gia Chungnam, Hn Quc.
Nui cy t bo: Cc t bo MC3T3-E1 c ly t ngn hng t bo
RIKEN (Tsukuba, Nht Bn) v nui cy nhit 37oC trong mi trng inh
dng ti thiu iu chnh (-MEM; GIBCO) trong iu kin khng kh cha
5% CO2. Thng mi trng c cha 10% huyt thanh phi b c hot ha
bng nhit (fetal bovine serum - FBS), 100 U/ml penicillin v 100 100 g/ml
streptomycin.
S pht trin ca t bo: Th nghim MTT c s dng xc nh
mc pht trin ca cc t bo MC3T3-E1. Cc t bo c nui cy mt
5x103 t bo/mt ging v nui cy trn phin 48 ging. Mc pht trin ca t
bo c xc nh sau 2 ngy tnh t thi im x l vi cc hp cht th nghim
s dng my o mu da trn s hp th mui 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl-tetrazolium bromide (MTT) ca cc t bo pht trin. Tm tt l: dung
dch MTT (0.5 mg/ml) c b sung vo mi trng nui cy cc t bo,
37C cho php s phn ct ca vng tetrazolium bng cc enzim
dehydrogenase ty th v s to thnh cc tinh th formazan mu xanh. Sau 2 gi,
lng MTT cn li c loi b mt cch cn thn v cc tinh th c ha tan

50

bng cch vi DMSO trong 20 pht. Phin c lc trong 5 pht v hp th

bc sng 570 nm c xc nh bng ph k.
Hot lc ca enzym Alkanine Phosphatase (ALP): Cc t bo c x l
vi mi trng cha 10 mM -glycerophosphate v 50 g/ml axit ascorbic,
khi u qu trnh khong ha in vitro. Mi trng c thay th nh k 2-3
ngy mt ln. Sau 6 ngy, cc t bo c nui cy vi mi trng c cha 3%
albumin huyt thanh b (bovine serum albumin - BSA) v tng hp cht th
nghim trong 3 ngy. Khi kt thc, mi trng c loi b v mng t bo c
ra nh 2 ln vi dung dch mui m phosphate (phosphate buffered saline PBS). Cc t bo c phn gii vi 0.2% Triton X-100, dch thy phn c ly
tm tc 14000 v/p trong 5 pht. Phn dch trong pha trn c s dng
xc nh hot lc enzim ALP s dng kit th hot lc enzym ALP c cung cp
bi cng ty Asan, Hn Quc.
Xc nh hm lng collagen: Hm lng collagen c xc nh bng
phng php so mu Sirius Red. Cc nguyn bo xng nui cy c ra vi
PBS, sau c nh bng dch Bouin trong 1 gi. Sau qu trnh c nh, dch c
nh c loi b v cc a nui cy c ra bng cch nhng vo dng nc
ang chy trong 15 pht. Cc a ny c kh tron khng kh v nhum bng
thuc nhum Sirius Red trong 1 gi di iu kin lc nh nhng. Sau , loi b
phn dch v mi trng nui cy c ra vi HCl 0,01N loi b phn thuc
nhum khng c lin kt. Ha tan mi trng nui cy c nhum mu
bng NaOH 0,1 N v cng hp th c o bc sng 550 nm (s dng
NaOH 0,1 N lm i chng).
Kh nng tch t canxi: Cc t bo c x l vi mi trng cha 10
mM -glycerophosphate v 50 g/ml axit ascorbic, khi u qu trnh khong
ha in vitro. Sau 10 ngy, cc t bo c nui cy trong mi trng cha 0,3%
BSA v tng hp cht th nghim trong 3 ngy. Khi kt thc, cc t bo c c
nh bng etanol 70% trong 1 gi, sau nhum bng thuc nhum 40 mM
Alizarin Red S trong 10 pht di iu kin lc nh nhng. nh lng cc
thuc nhum lin kt, thuc nhm c ha tan bng cch lc vi
cetylpyridinum chloride 10% trong 15 pht iu kin khng c nh sng.
Cng hp th c xc nh bc sng 561 nm.
Tnh ton: Kt qu c biu th bng meanSEM. Phn tch thng k
c thc hin bng phn mm ANOVA (P<0.05) vi h thng SAS.
e. nh gi hot tnh khng NF-B.
Cc th nghim hot tnh khng NF-B c tin hnh theo phng php
SEAP ASSAY trn dng t bo chun RAW264 ti Phng nghin cu ung th,
Vin Sinh hc v Cng ngh sinh hc Hn Quc (KRIBB). Phng php th
nghim c trnh by tm tt nh sau:
Dng t bo RAW264 c nui trong mi trng DMEM (Dulbecco
Modified Eagles Medium) c cha 0,5% huyt thanh phi b n khi t 2x105
t bo/ml, ly 180 l vo mi ging ca phin vi lng 96 ging, sau 3 gi ri
b sung 2 l mu th cc nng khc nhau trong dimethylsulfosit (DMSO) v
20 l dung dch DMEM ni trn cng vi cht hot ha LPS (Lipopolysacharide)
theo t l 1 l LPS/1ml DMEM ri trong 24h, sau chuyn 100 l dung dch
51

trong mi ging sang ging ca phin 96 ging mi, a ny 65oC trong 10

pht, thm 100 l dung dch m 2xSEAP (Secretable Alkaline Phosphatase) vo
mi ging, tip tc 37oC trong 10 pht, trong qu trnh , chun b c cht (pnitrophenol phosphate, 31,6 mg/ml) vi dung dch m 1xSEAP ri thm 20 l
dung dch c cht vo mi ging, 37oC trong 2h, sau o cng hp th
mu bc sng 405 nm bng my Microplate Reader (Molecular Devices Co.,
Menlo park, CA, USA). Cht i chng c s dng trong cc th nghim ny
l Parthenolide.
II.2.3. Phng php phn lp v xc nh cu trc ho hc
Cc phng php v thit b nghin cu ch yu gm:
a. Sc k lp mng (TLC)
Sc k lp mng c thc hin trn bn mng trng sn silica gel DCAlufolien 60 F254 (Merck 1.05715).
Pht hin vt cht: n UV 254 v 368 nm, phun H2SO4 10% ln bn mng
ri h nng t t n khi hin mu v th Dragendoff.
b. Sc k ct (CC)
Dng loi silica gel 230-400 mesh (0,04-0,063mm) Merck. ODS-R18
Merck.
c. Ph hng ngoi (IR)
Ph hng ngoi (p vin vi KBr): o trn my Jasco 200 (Vin Ho hc,
Vin Khoa hc v Cng ngh Vit Nam).
d. Ph khi lng (MS)
Ph khi lng (ESI-MS) c o trn my LC-MSD Agilent 1200 Series
(USA) ca Vin Ha hc cc Hp cht Thin nhin, Vin Khoa hc v Cng
ngh Vit Nam.
Ph khi lng phn gii cao (HR FAB-MS) c o trn my JEOL JMSDX 300 ca Vin Nghin cu khoa hc c bn Hn Quc (KBSI).
e. Ph cng hng t ht nhn (NMR)
Ph NMR o trn cc my: Bruker AM300; Bruker AM600 ca Vin
Nghin cu khoa hc c bn Hn Quc (KBSI); v my AVANCE 500 ca Vin
Ho hc, Vin Khoa hc v Cng ngh Vit Nam. Cht ni chun l TMS
(Tetrametyl Silan).

52

nh mt s thit b s dng trong cc phng php nghin cu

thu mu, chit tch v xc nh cu trc.

Mt s thit b s dng ly mu v gim nh tn khoa hc

Mt s thit b s dng chit xut v phn lp cc hot cht

H thng ph cng hng t ht nhn (NMR) v ph khi lng (ESI-MS) s dng

xc nh cu trc cc hot cht

53

II.2.4. Nguyn liu v phng php nghin cu thm d kh nng khng

vim t rong to bin Vit Nam.
II.2.4.1. Nguyn liu
a. Mu rong:
Danh sch 22 mu rong to bin c s dng trong nghin cu c trnh
by trn bng II.2.4.1 do cn b Phng Cng ngh To, Vin CNSH v CN.
Hunh Quang Nng, Vin nghin cu v ng dng Cng ngh Nha Trang
(VAST) thu thp v nh tn khoa hc. Thi gian thu mu 2007-2008. Trong ,
Undaria pinnatifida, Enteromorpha linza l hai loi c hot tnh khng vim cao
do Trng i hc Quc gia Pukyong, Hn Quc cung cp c s dng lm
mu th so snh vi rong bin Vit Nam.
Bng II.2.4.1. Danh sch 22 mu rong bin c dng tch dch chit c hot
tnh khng vim
TT

Tn mu

Gracilaria asiatica

G. blodgettii

Sargassum
duplicatum
Ulva reticulata (1)
Sargassum
polycystum
Codium arabicum
Enteromorpha
compressa
Ulva reticulata (2)

4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

Dictyota dichotoma
Sargassum
microcustum
S. mcclurei
S. denticarpum
S. swartzii
S. binderi
Gracilaria arcuata
G. eucheumoides
G. salicornia
Hypnea valentina
Porphyra crispata
P. suborbiculata
Undaria pinnatifida
Enteromopha linza

a im thu mu
m nui trng thu sn, Trm Nghin cu nui
trng thu sn nc l Qu Kim, Hi Phng
m nui trng thu sn, Trm Nghin cu nui
trng thu sn nc l Qu Kim, Hi Phng
Ca l, Ngh An
Do Cng ty TNHH Vit i cung cp
Do Cng ty TNHH Vit i cung cp
Nha Trang, Khnh Ho
Nha Trang, Khnh Ho
Nha Trang, Khnh Ho
Nha Trang, Khnh Ho
o tri Nguyn, Nha Trang, Khnh Ho
Hn Chng, Vnh Phc, Nha Trang, Khnh Ho
M Ho, Ninh Hi, Ninh Thun
Bi Tin, Vnh Ho, Nha Trang, Khnh Ho
Tri Bu, Vnh Nguyn, Nha Trang, Khnh Ho
Bi Tin, Vnh Ho, Nha Trang, Khnh Ho
Nha Trang Khnh Ho
Nha Trang Khnh Ho
Nha Trang Khnh Ho
Tha Thin Hu
Tha Thin Hu
i hc Quc gia Pukyong, Hn Quc
i hc Quc gia Pukyong, Hn Quc

54

b. ng vt th nghim
+ Chut cng trng (c v ci) kho mnh c trng lng khong 120
20g do Trung tm chn nui ca Hc Vin Qun Y cung cp.
+ Chut nht c c trng lng khong 22-25 gam do Vin V sinh dch
t Trung ng cung cp (Hnh II.2.4.1).
Cc ng vt th nghim c nui trong iu kin ng nht ca phng th
nghim c y thc n v nc ung trc khi dng lm th nghim.

Hnh II.2.4.1. Chut nht c c s dng trong th nghim

xc nh hot tnh khng vim ca rong bin
c. Cc chng vi sinh vt v nm gy bnh kim nh
Cc chng vi sinh vt c dng th nghim hot tnh khng vi sinh vt
kim nh bao gm: Escherichia coli; Pseudomonas aeruginose; Bacillus
subtilis; Staphylococcus aureus v nm Candida albicans
d. Ho cht
Axit arachidonic (AA) (hng Sigma), Metanol, Aceton, Ethanol,
Acetonitrile, Benzene, Chloroform, Cyclohexan, Ethyl axetat, Ethyl alcohol,
Hexan, Methyl alcohol, Ete (Trung Quc), carrageenin, formaldehyd v cc ho
cht khc c s dng trong th nghim. Cht i chng: Amphotericin B,
Amoxicilin, natricloid morphin clohydrat ng 10 mg/1 ml, indometacin v
aspergic loi gi 100 mg.
e. Dng c
Km bm ng knh 0,5cm (c), thc kp VERNIER CALIPER
(Trung Quc), my nh k thut s Canon ICY Digital 70 (Nht Bn), Thit b
ct quay ROTAVAPOR-R (hng BUCHI, Laborotorums Technikz-AG Thu s),
Pipt cc loi (hnh II.2.4.2.a-b), my huyt hc t ng ABC (Animal Blood
Counter) ca hng Ugo- Basile, Italy, my xt nghim sinh ho mu t ng
Screenmaster ca hng Hospitex Diagnostic, Italy, my hot-plate ca hng UgoBasile (Italy), ph k Plethysmometer No 7250 ca hng Ugo - Basile (Italy).

55

II.2.4.2. Phng php nghin cu

a. Phng php phn tch lipid tng s
Theo tiu chun ISO/DIS 659:1998, LB c.
- Mu sau khi cn tng khi lng em ngm chit trong 100 mL
clorofoc/metanol (1/2);
- Lc, loi b thu dch chit I;
- B sung 100 mL nc vo dch chit tng, lc k to dch chit II;
- dch chit II phn lp trit , chit ly pha di v lm khan bng Na2SO4;
- Ct loi dung mi di p sut thp thu lipid tng, cn khi lng.
b. Phng php phn tch thnh phn axit bo
Theo tiu chun ISO/FDIS 5590: 1998, LB c.
- Trong thc nghim 10 mg du bo c ha tan vi 1mL n-Hexan, lc k
trong l nt kn;
- B sung 25 l dung dch CH3ONa trong metanol (2 Mol/L) v lc k
trong 1 pht;
- B sung 1 mL H2O ct vo, lc k v phn lp bng li tm 3000
vng/pht. Ht lp sp khng phn ng di loi i;
- B sung 100 l HCl, lc k v phn lp bng li tm 3000 vng/pht;
- Loi b kit lp bn di y, lp dung mi trn c lm khan bng
Na2SO4 v phn lp bng li tm 3000 vng/pht;
- Chuyn mu methyl ha sang ng mu em phn tch thnh phn axit
bo bng my sc k kh: HP-6890, ghp ni vi Mass Selective Detector Agilent
5973; Ct: HP-5MS (0.25(m* 30m*0.25mm); Kh mang He; Chng trnh nhit
: 80(1min.) 40/min. 150 (1min) 10/min 260 (10min.). Th vin ph
khi: WILEY275. L v NIST 98.L (Ph v iu kin phn tch my GC-MS km
theo).
a. Quy trnh tch chit dch c hot tnh khng vim:
Dch chit c hot tnh khng vim trong cc mu to nghin cu c
tch chit theo phng php ca Jin v cs, 1997 [141] v Khan 2007 [142].
b. Phng php th hot tnh khng vim trn tai chut
Theo phng php ca Van Arman (1974) [143]. Th nghim c chia
thnh 2 l: i chng v th nghim, mi l c lp li t 6-7 con
Trong l i chng (C): - Tai tri ca chut c bi 5l dung dch 100mg
AA/ml ho tan trong acetone - cht gy nhn to vim.
- Tai phi - 5l dung dch axeton.
Trong l th nghim (TN): - Tai tri ca chut c bi 5l dung dch 100mg
AA/ml v 5l dch chit cc loi rong bin nghin cu c nng 0,4 mg/5l.
- Tai phi ca chut bi 5l dung dch AA 100mg/ml
- X l kt qu: Hiu qu ca cc dch chit khng vim, c nh gi qua
cc gi tr ph n (c xc nh bng trng lng v dy tai chut - W) v
ban (l phn trm mu ti (M-%) trn tai chut xut hin sau khi gy ph
n nhn to bng AA v s gim gi tr ph n nu trn bng vic bi dch chit
c hot tnh khng vim gia l th nghim v i chng, sau chp nh v
phn tch bng phn mm Adobe Photoshop (CS 8.0).
56

Trong :
* Gi tr ph n = W tai phi ca TN - W tai tri ca TN.
Phn trm c ch ph n = [(Wtai phi ca TN - Wtai tri ca TN)/(Wtai tri ca C - Wtai phi ca C )]*100.
* Gi tr ban = M tai phi ca TN - M tai tri ca TN.
Phn trm c ch ban = [(Mtai phi ca TN - Mtai tri ca TN)/(Mtai tri ca C - Mtai phi ca C )]*100.

Hnh II.2.4.2.a. Dng c v thit b c s dng: A/ Thit b ct ROTAVAPORR, B/ thc kp VERNIER CALIPER, C/ Km bm ng knh 0,5cm

Hnh II.2.4.2.b Dng c chup nh phn tch ban (A) v dng c ct tai
xc nh trng lng tai (B)
R: Aceton

L: AA

R: AA +EtOH

i chng

L: AA+ Dch chit

Th nghim

Hnh II.2.4.2.c. S th nghim xc nh hot tnh khng vim ca dch chit

tch t cc loi rong bin nghin cu trn tai chut

57

c. Phng php th hot tnh khng vi sinh vt kim nh (pha long nng )
- Pha long mu th bng phng php pha long a nng : Mu ban
u c nng 20mg/ml c pha long thnh cc nng khc nhau th
hot tnh vi cc chng c nng 128 g/ml; 32 g/ml; 8 g/ml; 2 g/ml; 0,5
g/ml i vi cht sch., 256 g/ml; 64 g/ml; 16 g/ml .. 1 g/ml; 0,25 g/ml
i vi dch chit.
- Th hot tnh
+ Chun b dung dch vi sinh vt hoc nm vi nng 5.105 trc khi tin hnh
th
+ Ly 10 l dung dch mu th theo cc nng c pha long thm 200 l
dung dch vi sinh vt hoc nm, 370C sau 24 gi o c t bo trn my
Tecan, x l s liu v tnh ton gi tr IC50. Cc th nghim ny c tin hnh
ti Phng th hot tnh sinh hc, Vin Ho hc, VAST.
d. Nghin cu c tnh cp
Nghin cu c tnh cp v xc nh LD50 ca dich chit c hot tnh
khng vim t Sargassum swartzii (S) v Ulva reticulata (U) trn chut nht
trng bng phng php ca Litchfield v Wilcoxon (1949) [144]. Chut c
nhn i 12 gi, chia thnh cc l khc nhau, mi l 10 con, ung to vi liu
tng dn xc nh liu thp nht gy cht 100% s chut v liu cao nht gy
cht 0%. Chut c theo di tnh trng chung trong sut 7 ngy v t l chut
cht trong 72 gi ca tng l chut sau ung thuc.
e. Nghin cu tc dng gim au
* Gim au bng phng php Koster - qun au axit axetic:
Chut nht trng c chia ngu nhin thnh 6 l, mi l 10 con
- L 1 (i chng m): ung nc ct 0,2 ml/10 g chut/
- L 2 (i chng dng): ung aspirin liu 100 mg/kg.
- L 3: ung dch chit S liu thp 250mg/kg.
- L 4: ung dch chit S liu cao 500mg/kg.
- L 5: ung dch chit U liu thp 250mg/kg.
- L 6: ung dch chit U liu cao 500mg/kg.
Cho chut ung thuc th 3 ngy lin vi th tch 0,2ml/10g. Ngy th 3,
sau khi cho chut ung thuc 1 gi, tim vo bng chut dung dch axit axetic
1%, 0,2 ml/con. m s cn qun au ca tng chut trong mi l tng 5 pht
cho n ht pht th 25 sau khi tim axit axetic.
* Phng php mm nng:
Chut nht trng c chia ngu nhin thnh 6 l, mi l 10 con
- L 1 (i chng m): ung nc ct, th tch 0,2 ml/10 g chut.
- L 2 (i chng dng): tim mng bng morphin liu 10 mg/kg.
- L 3: ung dch chit S liu thp 250mg/kg.
- L 4: ung dch chit S liu cao 500mg/kg.
- L 5: ung dch chit U liu thp 250mg/kg.
- L 6: ung dch chit U liu cao 500mg/kg.

58

Th nghim c tin hnh ti hai thi im: trc v sau khi cho chut
ung thuc th 3 ngy (nh phn th nghim trn). t chut ln mm nng, lun
duy tr nhit 56oC bng h thng n nhit. Tnh thi gian t lc t chut
vo mm nng n khi chut lim chn sau. Loi b nhng chut phn ng trc
8 giy v sau 40 giy.
* Phng php gy vim cp
+ Gy ph chn chut bng carrageenin.
Chut cng trng c chia ngu nhin thnh 6 l, mi l 10 con.
L 1 (i chng): ung NaCl 9, 1 ml/100 g.
L 2: ung indometacin liu 25 mg/kg.
L 3: ung dch chit S liu thp 175mg/kg.
L 4: ung dch chit S liu cao 350mg/kg.
L 5: ung dch chit U liu thp 175mg/kg.
L 6: ung dch chit U liu cao 350mg/kg.
Chut c ung thuc 3 ngy lin tc trc khi gy vim. Ngy th 3,
sau khi ung thuc th 1 gi, gy vim bng cch tim carrageenin 1% (pha
trong nc mui sinh l) 0,1 ml/chut vo gan bn chn sau, bn phi ca chut.
o th tch chn chut (n khp c chn) bng dng c chuyn bit vo cc thi
im: trc khi gy vim (V0); sau khi gy vim 2 gi (V1), 4 gi (V2), 6 gi (V3)
v 24 gi (V4).
Kt qu c tnh theo cng thc ca Fontaine.
+ tng th tch chn ca tng chut c tnh theo cng thc:

V% =

Vt V0
100
V0

Trong : V0 l th tch chn chut trc khi gy vim

Vt l th tch chn chut sau khi gy vim
+ Tc dng chng vim ca thuc c nh gi bng kh nng c ch phn ng
ph (I%)
I% =

V c % V t %
100
V 0 %

Trong : V c % : trung bnh tng th tch chn chut l i chng

V t % : trung bnh tng th tch chn chut l ung thuc
+ Gy vim mng bng
Chut cng trng c chia ngu nhin thnh 6 l, mi l 10 con.
L 1 (i chng): ung NaCl 9, 1 ml/100 g.
L 2: ung indometacin liu 25 mg/kg.
L 3: ung dch chit S liu thp 175mg/kg.
L 4: ung dch chit S liu cao 350mg/kg.
L 5: ung dch chit U liu thp 175mg/kg.
L 6: ung dch chit U liu cao 350mg/kg.
Cc l chut c ung thuc th hoc nc mui sinh l vi thi gian
nh th nghim trn. Gy vim mng bng bng dung dch carrageenin 0,05 g +

59

formaldehyd 1,4 ml pha trong 100 ml nc mui sinh l va , vi th tch 2 ml

vo khoang mng bng cho mi chut.
Sau 24 gi gy vim, m bng chut ht dch r vim. o th tch, m s
lng bch cu/ml dch r vim v nh lng protein trong dch r vim.
* Gy vim mn (gy u ht)
Chut nht trng, c chia ngu nhin thnh 6 l, mi l 10 con. Cc l
c b tr th t nh th nghim phn gy trn dch mng bng. Gy vim mn
tnh bng cch cy si amiant trng lng 6 mg tit trng (sy 120oC trong 1 gi)
c tm carrageenin 1%, da phn gy ca mi chut.
Sau khi cy u ht, cc chut c ung NaCl 9 hoc thuc th lin tc trong 5
ngy. Ngy th 6 tin hnh git chut bng chlorofoc, bc tch khi u ht, sy
kh nhit 56oC trong 18 gi. Cn trng lng u ht sau khi c sy kh.
Tt c cc th nghim nu phn d v e c thc hin ti B mn Dc l,
Trng i hc Y H Ni.
f. X l s liu
Cc s liu nghin cu c x l thng k theo phng php t-test Student
v test trc - sau(Avant - Aprs). S khc bit c ngha khi p < 0,05.
II.2.5. Nguyn liu v phng php nghin cu thm d kh nng sinh cc
cht c Hot tnh sinh hc t vi sinh vt bin
II.2.5.1. Nguyn liu
- Cc mu nc v trm tch c ly cc a im khc nhau ca bin Vit
Nam: Qung Bnh, Bnh nh, Ph Yn vo thng 5/2007. Cc mu ny c
Vin a L cung cp.
- Cc mu san h v rong ly t bin Nha Trang, o Trng Sa 6/2007 do
Phng Sinh thi nc Trung tm Nhit i Vit Nga cung cp. Cc mu nc
bin, trm tch bin su > 2m tr ln c ly bng cc dng c chuyn
dng.
- Cc mu nc b mt bin Hi Phng, Ct B, Nha Trang, H Long, Nam nh
do nhm ti thu thp thng 6-7/2007.
- Cc mu c bin, ngao, s huyt mua ti ch Thnh Cng, H Ni vo thng
5/2007.
- Su t (Plutella xylostella), su xanh (Heliothis armigera) do Vin Bo v Thc
vt cung cp.
- Giun nhiu t Derodigitata miiler (thc n cho c cnh) mua ti ch H Ni.
- 5 chng VSV kim nh nhn t bo tng ging chun ca Trung tm CNSH,
i hc khoa hc t nhin H Ni: vi khun Gram (-): E. coli; Pseudomonas
aeruginose, vi khun Gram (+): Bacillus subtilis; Streptococcus aureus, Nm
men gy bnh: Candida albicans.
II.2.5.2. Mi trng phn lp vi sinh vt (g/l)
- Mi trng MPA (phn lp vi khun) Cao tht: 5; pepton: 10; NaCl: 5; Thch:
20; Nc bin 100%.

60

- Mi trng Gause 1 (phn lp x khun) Glucoza :10; K2HPO4: 0,5; MgSO4:

0,5; KNO3: 1; FeSO4: 0,01; Thch: 20; Nc bin.
- Mi trng Hansen (phn lp nm men) Glucoza: 50; pepton 5; MgSO4.7H2O:
3; K2HPO4: 3; KH2PO4: 3; Cao nm men: 1; Thch: 20; Nc bin.
- Mi trng Czapek (phn lp nm mc) Glucose: 5: MgSO4: 0,5; KCl: 0,5;
MgCl2: 3; FeSO4: 0,01; NaNO3: 3; KH2PO4: 1; K2HPO4 : 1; Thch: 20; Nc
bin. Tt c cc mi trng trn u c b sung dung dch nguyn t vi lng.
II.2.5.3. Phng php nghin cu
- S lng vi sinh vt trong cc mu phn tch c xc nh theo phng
php pha long ti hn ca Koch. [145]. Cc chng vi sinh vt c tch, nh k
hiu da vo c im ca mu v hnh thi, mu sc, kch thc ca khun lc.
Cc chng phn lp t nc bin k hiu N, phn lp t trm tch bin k hiu T,
t rong to bin k hiu R v t ng vt bin k hiu C.
- Sng lc s b kh nng sinh cht khng khun, khng nm ca cc
chng phn lp c trn a thch MPA cha 2% dch VSV kim nh. Sy kh
mt thch trong bung cy 45 pht. Chia a thch ra thnh 49 vung. Mi
cy gt trc tip 1 chng VSV. t cc a thch vo t m 370C. Sau 48 ting ly
ra kim tra. Nu chng no sinh cht khng khun, khng nm th to vng trong
xung quanh vt cy. m s chng to vng khng khun, khng nm v tnh t
l % s chng c kh nng khng khun trn tng s chng VSV th nghim.
Cy cc chng c vng v khun vo ng thch nghing cho cc th nghim
th hot tnh sinh hc tip theo.
- Nhum Gram: lm tiu bn cc chng vi khun c hot tnh sinh hc.
Thuc nhum l tm kt tinh, Iodinmordant, cn 95%, safranin 5%. Cc mu
c quan st di knh hin vi quang hc bng vt knh du. Vi khun Gram (+)
bt mu xanh-tm, vi khun gram (-) mu , hng.
- M t hnh thi khun lc bng phng php cy khun lc ln trn a thch
[145].
- Hot ho cc chng vi khun kim nh trn mi trng MPA 370C
trong vng 24 gi, cn chng nm men C. albicans trn mi trng Hansen
300C trong 48 gi.
- Nui cy VSV bin th kh nng sinh cc cht c HTSH: nui cy lc
200 vng/pht trn cc mi trng c b sung nc bin thch hp vi tng nhm
VSV. Thu dch nui cy vi khun sau 2 ngy, dch nui cy nm si, x khun
sau 7 ngy v nm men sau 5 ngy. Sau ly tm b loi cn.
- Xc nh kh nng khng cc VSV kim nh bng phng php c l
v o vng v khun [145]. Kh nng khng khun v nm gy bnh c xc
nh bng hiu s ng knh vng v khun to thnh trn a thch v ng
knh ca l thch (d1-d2). Chng no c ng knh vng v khun >17mm
(tng ng vi 20 n v khng sinh quc t IU) th c chn nghin cu
tip.
- Phng php xc nh kh nng gy c t bo trn giun t Derodigitata
miiler: dch nui cy cc chng VSV bin c kt ta bng butanol, t l 1:1 v

61

sy kh. Sau cn c pha trong 5ml nc ct. Chun b 10 ng nghim cha

10ml nc my, cho 50 con giun t vo mi ng nghim. B sung vo 5 ng th
nghim (TN) 0,5, 1,0 v 1,5ml dung dich nui cy. B sung nc my vi cc
liu lng nh trn vo 5 ng i chng (C) Th nghim c lp li 3 ln.
Quan st v m s lng giun t cht sau 45 pht v tnh t l %.
- Phng php th kh nng dit su hi: Su t v su xanh nui 250C
trn a Petri, mi cng thc 30 con su, liu lng dch th 1,0 ; 1,5 ; 2,0ml. L
TN: thc n v su c phun dch nui cy VSV bin. C: thc n v su c
phun nc ct. Theo di trong 5 ngy. Quan st v m s lng su cht sau 5
ngy.
- Xc nh hot tnh amilaza trn mi trng cha 1% tinh bt tan; hot
tnh xenluloza trn mi trng cha 1% bt giy v hot tnh proteaza trn mi
trng cha 1% cazein. Dung dch hin mu l Lugol 1%.
-Mi trng nui cy v thu sinh khi xc nh thnh phn axit bo khng
no l ng knh saccharoza 2%, cao nm men 0,5%, nc bin 20%. Mi trng
nui cy v thu sinh khi xc nh thnh phn axit bo khng no l ng knh
saccharoza 2%, cao nm men 0,5%, nc bin 20%. Nui 24 gi trn my lc
200 v/pht 280C. Trng lng kh xc nh bng phng php sy phun. T bo
nm men c ph v bng p lc 30.000 psi trn my ph t bo khong 30
pht trong dung dch m mui photpht natri axit citric 0,1M, pH = 5,0. Thnh
phn axit bo c xc nh bng my sc k kh 17A ca hng Shimadzu. Ct
Supelco Omegawax TM 250, di 30m, ng knh 25mm. Nhit bung bm
mu 2400 C, nhit ct l 1800C. Kh mang l He.
- Xc nh nh hng ca ngun cacbon, nit, pH v thong kh ln
sinh trung v sinh tng hp cc cht khng khun, khng nm theo phng php
VSV thng dng [145].
- nh tn khoa hc cc chng vi sinh vt bin c hot tnh sinh hc theo
phng php hnh thi, sinh l sinh ho v sinh hc phn t.
II.2.6. Nguyn liu v phng php nghin cu to thc phm chc nng b
sung omega-3 phng v h tr iu tr bnh tim mch
II.2.61. Phng php nghin cu
a. Phng php cng ngh
- Phng php p n trn my p n VTP 2006 do Vin Cng nghip Thc
phm thit k.
b. Phng php phn tch ho l
- Xc nh hm m theo phng php sy n 1050C trong 3 gi n trng
lng khng i [146].
- Xc nh m ton phn bng phng php Kieldahn [146].
- Xc nh hm lng bo bng phng php Sochlex [146].
- Xc nh n: n ca bng c xc nh bng t s gia din tch ca
bng/din tch u khun [147].
- Xc nh mu (L, a, b) bng my Hunter M.
- Xc nh nht trn my o nht Brookfield DV-II.

62

Hm lng DHA c xc nh ti Vin Dinh dng.

Xc nh axt bo omega 3 trn my sc k kh: cht bo c methyl ho,
chy trn my GC PLUS HP 6890 ca Vin Nghin cu Dc phm v M
phm min Nam.
c. Phng php vi sinh [148]
- Xc nh tng s vi khun hiu kh trn mi trng thch tht, pepton.
- Pht hin E. coli trn canh thang thng, canh thang lactoza, nc pepton.
- Pht hin vi khun Salmonella trn mi trng EMB (eosin methylen blue).
- Pht hin nm mc, nm men trn mi trng thch malt.
d. Phng php th nghim lm sng
i tng th nghim
i tng th nghim l cc bnh nhn c cc yu t v nguy c ca bnh
tim mch nh cao huyt p, ri lon chuyn ho lipit mu ang c iu tr
ti Bnh vin a khoa tnh H Ty.
Nam:
21 ngi
N:
27 ngi
S ngi < 45 tui: 1 ngi
S ngi > 45 tui: 47 ngi
Tin s bnh tim mch, tng huyt p:
< 1 nm:
4 ngi
1 2 nm:
13 ngi
2 5 nm:
22 ngi
5 10 nm:
6 ngi
> 10 nm:
3 ngi
Phng php theo di
Bnh nhn c chia thnh 2 nhm
- Nhm i chng (24 ngi): khng n thc n chc nng
- Nhm th nghim (24 ngi): n thc n chc nng, s lng 02
gi/ngy (mi gi 40 gam), chia lm 2 ba: sng (7 8 gi), chiu (16 17 gi),
ba chiu n gim 1 bt cm. Thi gian s dng lin tc trong 2 thng.
Trc khi tin hnh th nghim, cc bnh nhn c khm lm sng: o
huyt p, lm in tm , o mch, lm xt nghim sinh ho bao gm chc nng
gan, chc nng thn, chc nng chuyn ho lipit mu. Sau mi thng kim tra li
mch, huyt p, in tm v cc xt nghim sinh ho nh gi hiu qu ca
vic s dng thc n chc nng cho ngi tim mch. ng thi, chng ti cng
theo di cc du hiu nh au bng, a lng, y bng, kh tiu, mn nga cc
bnh nhn trong nhm th nghim.
e. Phng php nh gi cm quan: theo TCVN [149]
f. Phng php ton hc
Qui hoch ton hc thc nghim ti u ha cc iu kin cng ngh ca
qu trnh p n bng ma trn ton hc thc nghim Doehlert.

63

II.2.6.2. Thit b, dng c, nguyn vt liu v ho cht s dng cho nghin cu

a.Thit b
- T sy SANYO (Nht Bn).
- T m SANYO (Nht Bn).
- T hp thanh trng TOMY (Nht Bn).
- My ly tm Sorvall RC6 (c).
- My c quay chn khng Buchi (Italia).
- My tch v (Vit Nam).
- My p n 1 trc VTP 2006 (Vit Nam).
- My nghin ba (Vit Nam).
- My trn (Vit Nam).
- My dp date (Trung Quc).
- My dn ti (Trung Quc).
- Cn phn tch Libro (Nht Bn).
- Cn k thut (Nht Bn).
- Knh hin vi Olympus LH 36 (Nht Bn).
-

b. Nguyn vt liu
Go Khang Dn, go lt mua ti Vin Khoa hc Nng nghip VN.
Bt tht mua ti Cng ty Thc phm Hng Sn.
u tng, vng trng mua ti th trng H Ni.
Malto dextrin mua ti Cng ty Minh Dng H Ty
Ch phm omega-3 do Vin Ha hc cc Hp cht Thin nhin cung cp.
c. Ho cht
Cao nm men
Pepton
Thch
Methylene xanh dung dch ru 0,2%
Methyl dung dch 0,1%
Phenolphtalein dung dch ru 1%
Axt sunfuric c (d = 1,84) v dung dch 0,1N
te etylic
Axetat
Methanol
Tetrahydrofuran
Natri cacbonat khan Na2CO3
Axt clohydric c (d = 1,19)
ng chun NaOH 1N
Natri hydroxyt dung dch 30%
Kali pemanganat dung dch 0,1N
St sunfat Fe2(SO4)3
ng sunfat tinh th CuSO4.5H2O
Kali sunfat tinh th K2SO4
V mt s ho cht khc

64

II.2.7. Phng php nghin cu to thc phm chc nng Bionamine tng
cng sc khe cho vn ng vin t sao bin
II.2.7.1. Nghin cu xc nh hm lng hot cht sinh hc hoocmn steroit
(testosteron, progesteron, cortilson, oestradoen) trong cc i tng nghin
cu theo phng php min dch Eliza trn my Multiskan - Phn Lan vi kit
ca hng BiotechM.
Phng php min dch ELISA: l phng php min dch enzym vi
khng th a dng c nhy rt cao v s tch t qua 1 pha rn. Cc hoocmn
steroit ca mu, ca chun v ca i chng phn ng vi khng th a dng
c nh du bng fluorescein trong ng th v cnh tranh vi dn xut ca
hoocmn steroit c nh du bng enzym gn vo s lng hn ch
khng th. Mt cht tch c bit s tch hoocmn steroit ra khi mi lin kt vi
SHBG v do lng hoocmn steroit tun hon tng s s c xc nh.
Lng dn xut hoocmn steroit c gn t l nghch vi lng hoocmn
steroit c trong mu.
Sau khong thi gian l 30 pht 370C, cht lng th khng li
flurorescein ( c gn vi cc phn t c th t tnh ho) s c cho thm
vo mc d tha (D). Cht khng th ny gn nhanh v c hiu vi phc hp
dn xut ca hoocmn steroit khng th v s lng xung trong 1 t trng m
khng cn phi ly tm, phn trn s c i v phn cn c ra. C cht
trong tt c cc ng c vi 1 dung dch c cht ca enzym (F). Sau khi 30
pht 370C th dng phn ng bng cch cho vo dung dch dng phn ng(G).
Cng ca mu sc ca ng c xc nh bng so mu, cng ca mu
sc xut hin do phn ng enzym t l nghch vi nng ca khng nguyn
ang tn ti. Vi mu chun s dng c ng cong chun v t c th xc
nh c nng khng nguyn cha bit c trong mu cn phn tch.
II.2.7.2. Nghin cu xc nh hm lng hot cht sinh hc, cc nguyn t
khong vi lng cn thit (Fe, Zn, Mn, Se) trong cc i tng nghin cu
bng phng php phn tch hunh quang tia X trn my k hiu XRF SEA2110 ca Nht.
Phng php phn tch hunh quang tia X l mt trong nhng phng php
hin i v tin tin, nguyn l ca phng php ny l dng ngun nng lng
phng x tia X chiu qua c cht mu cn phn tch v lm cho cc cht c trong
mu pht ra nhng tia X c trng cho tng nguyn t. Mi nguyn t li pht
quang ra nhng im to ra mt ngun nng lng v khi so vi cc im chun
ca n ta c th tnh ra c hm lng ca tng cht .
II.2.7.3. Nghin cu xc nh hm lng hot cht sinh hc (protein, axit
amin) ca cc i tng nghin cu bng phng php in di trn my axt
amin t ng HP Amino Quant Series II ca Nht.
II.2.7.4. Nghin cu quy trnh khai thc cc hot cht sinh hc t to ch
phm thc phm chc nng vin tng lc Bionamine bng cng ngh enzyme.

65

* Phn tch hot cht sinh hc trong mt s loi sao bin bng phng php
sinh ha, phng php min dch Eliza, phng php vi sinh v phng php
phn tch hunh quang tia X.
* ng dng cng ngh enzyme, cng ngh ha sinh to cc sn phm bt
nguyn liu Bionamine.
* Phng php xc nh mc thu phn c cht (hay phng php xc nh
hm lng m ca mu b thu phn). Hin nay c rt nhiu phng php xc
nh hot tnh ca protease c p dng nc ta nh: Phng php Auson ci
tin, phng php Gross- fuld, phng php chun formol,... Trong ,
phng php Auson ci tin c dng rng ri nht, da trn c s xc nh
lng Tyrozin gii phng trong qu trnh thu phn protein. Phng php ny c
nhy cao, d thc hin nhng li c hn ch cn bn l: c nhng protein trong
qu trnh thu phn khng gii phng ra Tyrozin hoc ngay c i vi mt s
protein c gii phng ra Tyrozin trong qu trnh thu phn th ngoi phn hu cc
lin kt peptit c to thnh vi s tham gia ca cc gc Tyrozin cc protease
cng phn hu c cc lin kt peptit khc khng to thnh bi cc gc Tyrozin.
V vy, s dng phng php Auson ci tin xc nh hot tnh s khng nh
gi ng thc cht hot ca enzim protease
Khi lin kt peptit b thu phn th mt trong cc sn phm to thnh s
cha nhm amin (-NH2) v s lng amin mi to thnh bng lng lin kt b
thu phn. Cho nn vic nh lng nhm - NH2 s phn nh chnh xc thc trng
hot tnh protease.
C nhiu phng php nh lng nhm - NH2, trong c phng
php nh lng nhm - NH2 bng s dng 2, 4, 6 - nitrobenzen - 1- sulporic axit.
Nhng ho cht ny hin nay rt kh kim Vit Nam. Nn chng ti s dng
phng php khc nh lng - NH2, l phng php chun Formol.
Nguyn tc ca phng php: Formaldehyde lng d tha d dng lin
kt vi cc nhm amin (- NH2) dn ti gii phng ra proton (H+)
+
NH3CHRCOO- + NH3CHRCOO NH2CHRCOO- + H+
NH2CHRCOO- + 2 HCHO (HOCH2)2 NCHRCOOPuton gii phng ra c chun trc tip bng NaOH cho n khi xut
hin mu ca phenolphtalein (pH ~ 8) th dng li.
II.2.7.5. Nghin cu c tnh cp v c tnh bn trng din ca bt
Bionamine.
Cc nghin cu thc nghim v c tnh cp v bn trng din c thc
hin ti Trung tm nghin cu c hc Hc Vin Qun Y.
* i tng nghin cu
Nghin cu c tnh c thc hin trn 110 chut nht trng c trng
lng 20 (2 gam tui trng thnh (8-12 tun tui). Tt c cc chut c
chn u nhanh nhn, kho mnh, khng mc cc d tt bm sinh.
Tng s chut nghin cu c chia thnh 5 l th nghim, mi l 12 con
xc nh t l sng cht cc mc liu khc nhau v 02 l mi l 25 chut

66

theo di nh hng c bn trng din. S phn chia chut vo mi nhm c

thc hin theo nguyn tc hon ton ngu nhin.
Trc v trong qu trnh thc nghim, tt c chut u c nui dng
trong cng mt mi trng vi mi iu kin ng nht.
hnh thnh dung dch nghin cu tin hnh ho tan ng u 600 mg
bt (tng ng 02 vin thuc) trong 1 ml nc ct. Nh vy, dung dch nghin
cu cha nng hot cht l 600 mg/ ml. Nghin cu c thc hin trn 05
mc liu ca bt Bionamine. Mc liu ti thiu c kho st l 3mg/g th trng
ng vt nghin cu (gp 125 ln liu s dng trn ngi) v mc liu ti a l
30mg/g th trng ng vt nghin cu (gp 1250 ln liu s dng trn ngi).
* Phng php nghin cu
Nghin cu xc nh liu c ca bt Bionamine c tin hnh theo
nguyn l ca phng php Kerber da vo t l sng, cht ca ng vt th
nghim khi c s tng liu. Liu c c xc nh theo ch tiu LD50 (liu gy
cht 50% ng vt thc nghim).
Nghin cu bn trng din tin hnh trong thi gian 03 thng. Trong
khong thi gian ny ng vt nghin cu b ung bt buc bt Bionamine mc
liu 15mg/g th trng (mc liu gp 625 ln liu s dng trn ngi).
ng xm nhp ca bt Bionamine vo c th chut l ng tiu ho.
Dch lng cha hot cht trong bt Bionamine c a vo d dy chut qua
ng ming theo nguyn tc ung bt buc thng qua vic s dng kim cong
u ty chuyn dng dng cho ng vt nh. Trong thc nghim chng ti dng
loi xilanh c th tch l 0,1 ml m bo chnh xc ca lng dch lng
hot cht cn a vo c th chut.
Tnh ton liu hot cht cn nghin cu sao cho th tch lng dung dch
cha hot cht trong bt Bionamine bm vo ng tiu ho chut khng vt
qu 0,5ml dch lng i vi mi con.
Nh vy, khi tin hnh nghin cu i vi trng hp liu cao hn, th
tch dung dch vt qu 0,5 ml, chng ti thc hin ng tc ung bt buc 02
ln trong ngy vi khong cch thi gian
ung gia cc ln khng di 4 gi.
Tin hnh quan st hnh vi ca nhng chut b ung p buc dch bt
Bionamine trong khong thi gian 24 gi lin tc.
Trong nghin cu c tnh bn trng din tin hnh cho ung dung dch
bt Bionamine hng ngy vo thi im c nh (8 gi sng mi ngy) trong
khong thi gian 03 thng lin tc.
II.2.7.6. Phng php th nghim tc dng ca vin tng lc Bionamine trn
ng vt thc nghim.
a. Th nghim tc dng ca vin Bionamine ln mt s ch tiu huyt hc ca
chut cng trng.
Th nghim 1: c trin khai trn chut cng trng gm 20 con c
kho mnh, thi gian thc nghim l 20 ngy chia lm 2 l c trng lng ng

67

u lc bt u phn l th nghim, khong 90- 100 g c chia thnh 2 l, l th

nghim (TN1) v l i chng (C 2)
- L 1: Th nghim (TN1): 10 con chut c nui bng khu phn n c bn v
c b sung vin Bionamine vi liu dng l 150mg/con/ngy.
- L 2: i chng (C2): 10 con chut c nui bng khu phn n c bn v
c b sung ch phm vin Bionamine gi (placebo) vi liu dng l
150mg/con/ngy.
b. Th nghim tc dng tng cng th lc v sc bn ca chut trong qu trnh
bi ln ca vin Bionamine.
Th nghim th 2: c trin khai trn chut nht trng gm 40 con
(20con ci, 20 con c) cc c th chut u kho mnh, c trng lng ng
u (25-30g/con) chia lm 3 l, mi l 10 con (5con ci, 5 con c). S chut
dng lm th nghim v i chng c ch n nn c bn ging nhau. Ring
chut dng lm l th nghim th c n b sung thm cc ch phm khc nhau:
- L i chng (C): 10 con chut (5chut c v 5 chut ci) c nui bng
khu phn n c bn.
- L th nghim (TN): 10 con gm (5chut c v 5 chut ci) c nui bng
khu phn n c bn ging nh l C v c b sung ch phm Bionamine vi
liu dng l 30mg/con/ngy.
Ch tiu c nh gi l thi gian chut bt u bi cho n lc chng mt mi
v chm ln u.
II.2.7.7. Phng php nh gi tc dng lm sng ca vin Bionamine trn
vn ng vin in kinh
nh gi tc dng ca ch phm vin tng lc Bionamine trn vn ng
vin. ti tin hnh th nghim nh gi tc dng v sc bn, sc nhanh
(phn x thn kinh) v sc mnh c bp trn vn ng vin ca cc i in kinh
nh sau:
Vn ng vin cc nhm thc nghim c dng vin tng lc Bionamine
v nhm i chng dng vin tng lc Bionamine placebo. Vi liu dng
4vin/ngy, ung trc ba n 10-15 pht. Cc nhm vn ng vin ca cc
nhm i chng v thc nghim c ch dinh dng v tp luyn nh nhau,
bao gm cc bi tp v sc bn, ch gio n tp nng v tp nh.
Trc v sau thi gian tp luyn 1 thng c kim tra v cc ch tiu v sinh
l, sinh ho.
Cc phng php s dng:
* ng dng phng php Y hc th thao nghin cu tc dng ca vin tng
lc Bionamine trn cc nhm vn ng vin ca 2 i c nhm i chng v th
nghim u c ch dinh dng v tp luyn nh nhau, bao gm cc bi tp v
sc bn, sc do, ch gio n tp ym kh, tp a kh.
- Phng php kim nghim b i, i chng v t i chng, ly mu
mu ca vn ng vin trc v sau thi gian tp luyn.
- Kim tra v cc ch tiu v chuyn mn, s phm, phn x tm l v cc
ch tiu sinh l, sinh ho v thnh tch thi u [150 - 156].

68

- Tt c cc mu sinh ho u thc hin o trn my o sinh ho VISUAL

v kt chun ca Hng BIOMRIEUX ca Php.
- Cc ch tiu huyt hc c o trn my Sysmexse-9500 Nht cho php
xc nh c 32 ch tiu huyt hc khc nhau.
- Cc ch tiu phn x tm l c o trn my SK2000 ti Vin Khoa hc
Th dc Th thao.
- Cc ch tiu nhp tim, oxy mch, VO2max, tiu th oxy, chuyn ho nng
lng, cng sut vn ng, gng sc ti a c o trn h thng Cosmed
K4B2 My ca Italia, l thit b hin i nht hin nay ca Vin Khoa hc Th
dc Th thao. Cosmed l thit b o t xa, c cu to tinh vi, gn nh, c th
truyn d liu t thit b phn tch ti ni thu nhn thng tin (Monitor mn
hnh vi tnh) vi khong cch 800m, nn Cosmed K4B2 c ng dng trong vic
kim tra vn ng vin khng ch trong phng th nghim m c trn hin
trng tp luyn nh sn vn ng, nh thi u. Vi phng php kim tra cc
thng s y sinh bng thit b Cosmed K4B2 l mt thit b hin i , c chnh
xc cao. Cosmed K4B2 hot ng theo nguyn l Knipping. l s kt hp gia
phng php kim tra chc nng ca h tun hon v h hp (Cardiospirometre)
vi phng php o cng sut (Ergomatre) din ra trong sut qu trnh thc hin
bi tp.
- Cc ch tiu thnh tch c cc hun luyn vin trc tip kim tra.
II.2.8. Nguyn liu v phng php nghin cu sn xut vin nang mm bo
ng HALIOTIS
II.2.8.1. Phng php nghin cu qui trnh bo ch vin nang Haliotis
a. Ni dung nghin cu
- Nghin cu la chn dc liu v xy dng qui trnh bo ch vin nang mm
Bo ng HALIOTIS thnh dng bn thnh phm v thnh phm
- Nghin cu xy dng tiu chun c s cho sn phm vin nang mm bo ng
HALIOTIS
- Kim nghim sn phm vin nang mm HALIOTIS theo tiu chn c s
xy dng.
- Tin hnh ng k sn phm di dng thc phm chc nng vi B y t xin
cp giy php lu hnh ton Quc.
b. Nguyn liu v phng php nghin cu:
+ Nguyn liu:
Tht bo ng c ly t vng bin Nha Trang do Trung tm nhit i Vit
Nga chi nhnh Nha Trang cung cp vi s lng 30 kg.
+ Cch x l:
- Bo ng sau khi thu hoch tch b v cng loi ht tp cht v ph tng, ra
sch x l bo qun ng kh.
- Thi lt mng, sy cho n kh
- Nghin thnh bt mn

69

+ Phng php nghin cu:

* Nghin cu la chn dc liu v xy dng qui trnh bo ch vin nang mm
Bo ng Haliotis:
- Bo ng c la chn t theo tiu chun qui nh ca Dc in c th:
T l thu phn khong 73%, 24,58% protit, 0,44% cht bo, 1,98% tro.
- Xy dng qui trnh bo ch vin nang mm Haliotis:
- Bo ng sau khi thu hoch tch b v cng loi ht tp cht v ph tng, ra
sch x l bo qun ng kh.
- Thi lt mng, sy cho n kh
- Nghin thnh bt mn
- Nh ho bng du u lnh thu c dng bn thnh phm
- ng nang mm Gelatine trn dy chuyn ng nang mm t tiu chun
GMP WHO
- Sy vin
- ng l hoc p v, dn nhn hon thin sn phm
- Kim nghim thnh phm
- Nghin cu
II.2.8.2. Phng php nghin cu c tnh cp
- Phng php xc nh c tnh cp ca thuc Nh xut bn Y hc, 1996.
- Hng dn km theo quyt nh s 371/BYT Q ngy 12/3/1996 v xc nh
an ton cho thuc c truyn ca B Y t
- General guidelines for methodologies on research and evaluational of
traditional medicine. World Health Organization 2000
- Principles and Methods of Toxicology A. Wallace Hayes, 1994.
ng vt v phng tin th nghim:
- Chut nht trng v th c hai ging kho mnh, t tiu chun th nghim,
chut cn nng 20g 2g; th cn nng 2,3 kg 0,2 kg.
- Cc ch s huyt hc c o trn my Autocounter 910
- Ch s sinh ha c xc nh trn my Vital Microlab 300
Lm tiu bn vi th v phn tch m bnh hc c thc hin ti Khoa Nghin
cu thc nghim - Vin Y hc c truyn Qun i.
II.2.8.3. Phng php nghin cu c tnh bn trng din
Mu th: Liu th: 2 vin/ kg/ ngy
Ct vin nang mm, p ra ht cht dch v thm nc, trn u thu c
hn dch c cha lng thuc tng ng 2 vin/ 5ml. Khuy trn u hn dch
mi khi ly cho th ung.
Chun b mu th hng ngy trc khi cho th ung.
B tr th nghim v tin hnh: Th nghim c tin hnh trn 20 th, chia lm
hai nhm chng v th, mi nhm 10 th.
- Nhm chng: ung nc ct 5,0ml/ kg/ ngy
- Nhm th : ung 5ml dch mu th chun b nh trn vi liu 0,3g/ kg/
ngy
Cho th ung hng ngy, lin tc trong mt thng.

70

Th c 2 nhm c nui dng trong cng iu kin v cng ch dinh

dng.
Theo di v nh gi: Theo di th hng ngy v mc tiu th thc n, kh
nng hot ng, tnh trng phn, lng.
Trc khi th nghim, xc nh ch s bnh thng ca th v cn nng, cc
du hiu ton thn, cc ch s v huyt hc, ho sinh chc nng gan, thn. Ht
thi gian th nghim, nh gi li th trn cc ch tiu nh trc th nghim. M
th quan st i th cc t chc gan, thn, phi, tim v hnh dng, mu sc, th
cht, tnh trng xung huyt...
So snh kt qu trc v sau th nghim ca nhm chng, nhm th, gia
nhm chng v nhm th theo phng php thng k y sinh hc.
II.2.8.4. Phng php nghin cu tc dng bi b c th trn lm sng
a. Cht liu nghin cu
Vin nang mm Haliotis bo ch t Bo ng c ng gi 170 mg/vin,
do khoa dc - Vin YHCT Qun i sn xut.
b. i tng nghin cu
Gm 45 bnh nhn (BN) t nguyn 40 tui
+ Tiu chun chn bnh nhn
- Bnh nhn 40 tui, khng phn bit gii, ngh nghip.
- Bnh nhn c chn on l suy nhc c th theo tiu chun chn on ca
CDC nm 1994.
- Theo YHCT bnh nhn thuc chng h lao th can thn m h: au u, hoa
mt, chng mt, tai , mt ng, hay qun, th trng gy; ngi mt mi, hay cu
gt, lng bn tay chn nng; ra m hi trm; au lng; mi gi; nhc trong
xng.
- Bnh nhn chp nhn dng thuc, tham gia cng trnh nghin cu, thc hin
nhng iu quy nh trong qu trnh nghin cu.
- Bnh nhn iu tr ni tr ti khoa lo khoa v iu tr ngoi tr ti phng khm
bnh vin YHCT Qun i.
+ Tiu chun loi tr
- BN ang dng thuc khc hoc suy thn, suy gan, suy tim, thiu mu nng,
bnh thn kinh thc tn
- BN khng hp tc nghin cu.
c. Phng php nghin cu
- Phng php nghin cu: Theo phng php th nghim lm sng so snh trc
v sau iu tr.
- Thit k nghin cu: 45 bnh nhn p ng tiu chun nghin cu. Thi gian
nghin cu 4 tun.
- Phng php dng thuc: dng vin nang mm bo ng 2 vin/ln x 2 - 3
ln/ngy x 30 ngy iu tr ni tr ti Khoa Lo khoa v iu tr ngoi tr ti
phng khm bnh - Vin YHCT Qun i.
- Tt c cc bnh nhn nghin cu u c khm xt t m v ni khoa, lm
bnh n c chn on theo YHCT v y hc hin i, c phiu theo di, thm

71

khm hng ngy theo di kt qu v ghi tc dng ph khng mong mun ca

thuc.
+ Cc ch tiu theo di v lm sng: c nh gi vo ngy D0, D30.
- Mc suy nhc thng qua thang im ca Bugard v Crocq.
- Ri lon tr nh thng qua thang im Wechsler (ti a 79 im).
- Th trng, trng lc c, huyt p, s ci thin v n v ng.
* Theo di c lc
- Mi bnh nhn c o c lc 2 ln vo ngy D0, D30, vo cng 1 gi;
- Dng c o c lc l lc k bp tay Lin X (DRL-90).
- Phng php o: Khi o ch dng c ca bn tay v cng tay khng dng c ca
cnh tay.
* Theo di th trng:
- Mi bnh nhn c o trng lng 2 ln vo ngy D0, D30.
- o th trng bng cn liga c chia n 100g.
Theo di thang im nh gi lm sng mc suy nhc Crocq -Bugard
- nh du nhng triu chng m bnh nhn mc phi trong 10 mc sau
- Cho im mi triu chng mc phi: t 0 (bnh thng) n 10 (trm trng nht).
- Ly im s cao nht ca triu chng trong mc lm im s cho
mc (t 0 n 10).
- Cng tt c s im ca 10 mc c tng s im suy nhc .
Cc triu chng ca bnh nhn c ci thin khi s im ca mi ln thm
khm sau gim i so vi ln thm khm trc .
* Theo di ri lon tr nh thng qua thang im ca Wechsler [157, 158]. Thang
im gm 7 cu hi, tng s im l 79.
Cch tnh im nh sau:
V d cu hi 1: nh gi chung (6)
1. Hi tui
2. Nm sinh
3. Tn cha m con ci
4. Ni (tnh, huyn, thnh ph)
Tr li ng mi cu hi ph tnh 1,5 im
Tr li sai mi cu hi ph: tnh 0 im.
Cc cu hi khc cng lm tng t nh cu hi 1.
Cc triu chng ca bnh nhn c ci thin khi s im ca mi ln thm
khm sau tng ln so vi ln thm khm trc .
* Theo di s ci thin v n ng
+ Theo di v n: Chia 3 mc :
n tt: n ngon ming, mau i, mi ba n 2 bt cm.
n bnh thng: n < 2 bt cm /ba.
n km: C cm gic y bng chm tiu, khng mun n, n < 1 bt/ba
+ Theo di v ng: nh gi cht lng gic ng bng thang im Pittsburgh
(PSQI ) ca Daniel J.Buyse v CS (1988) [157].
Chia 3 mc :

72

- Ng tt: D ng, ng su, thi gian ng t 5-7 h/ngy, ng dy ngi thoi mi,
d chu.
- Ng bnh thng: 3h < thi gian ng < 5 h/ngy.
- Ng km: Ng tht thng, kh ng, d tnh gic, hay m, thi gian ng mi
ngy < 3h. Ng dy khng cm thy thoi mi.
+ Cc ch tiu theo di v cn lm sng: c nh gi vo ngy D0, D30.
- Bn c lm xt nghim (XN) 2 ln vo ngy D0 v D30: XN huyt hc
(cng thc mu: s lng hng cu, bch cu, hemoglobin) v XN sinh ha
(protid mu, cholesterol ton phn, chc nng gan, thn).
- Theo di tc dng khng mong mun ca thuc trn lm sng: bun nn, nn,
a chy, y bng, mn nga.
+ X l s liu: cc s liu c x l theo phng php thng k y - sinh hc.
II.2.9. Phng php nghin cu sn xut thc phm chc nng vin nang
mm hi sm HASAMIN
II.2.9.1. Phng php xc nh cc thnh phn dinh dng, hoocmon v
khong cht trong Vin nang hasamin
a. Nghin cu xc nh hm lng hot cht sinh hc hoocmn steroit
(testosteron, progesteron, cortilson, oestradoen) trong cc i tng nghin cu
theo phng php min dch Eliza trn my Multiskan - Phn Lan vi kit ca
hng BiotechM.
b. Nghin cu xc nh hm lng cc nguyn t khong vi lng cn thit
(Fe, Zn, Mn, Se) trong vin Hasamin bng phng php phn tch hunh quang
tia X trn my k hiu XRF SEA-2110 ca Nht.
c. Nghin cu xc nh hm lng hot cht sinh hc (protein, axit amin) ca
trong vin Hasamin bng my axt amin t ng HP Amino Quant Series II ca
Nht.
II.2.9.2. Phng php th nghim c tnh cp v c tnh bn trng din
c tnh cp v c tnh bn trng din ca vin nang mm hi sm
Hasamin c nghin cu ti Vin Y hc C truyn Qun i theo cc phng
php c trnh by mc II.2.7.2 v II.2.7.3.
II.2.9.3. Phng php th nghim tc dng ca vin Hasamin trn ng vt
i tng:
Th nghim tin hnh i vi 10 th c thuc ging Newzealand, nui ti
Trung tm Nghin cu D v Th Sn Ty. Th ging c trng lng 3,0 - 5,0
kg, tui 12 - 26 thng, kho mnh, giao phi thun thc, nui nhng chung
ring bit, ch nui dng bao gm thc n tinh hn hp 16% protein
80g/ngy v 600g thc n xanh.
Phng php:
Ly tinh th bng m o gi, nhit m o gi 45oC, ly tinh vo bui
sng. Th tch tinh dch (V), hot lc tin thng ca tinh trng (A), nng tinh
trng (C), sc khng (R) i vi dung dch NaCl 1% ca tinh trng xc nh theo

73

phng php ca Milovanov (1962), tng s tinh trng tin thng ca mt ln ly

tinh (V.A.C) xc nh theo phng php ca Herrick v Self (1962), t l tinh
trng sng (LS) theo phng php nhum mu phn bit vi eosin 5%, t l tinh
trng k hnh (K) theo phng php nhum mu vi fuxine, pH tinh dch o bng
giy o pH ca Trung Quc [151].
M hnh th nghim:
L 1: 05 con (s tai:4182; 813; 853; 833; 91531), th n theo khu phn c
s + HASAMIN.
L 2: 05 con (s tai:94514; 11558; 1471; 1000; 836), th n theo khu phn
c s + vin Placebo (Vin trng)
Thi im nh gi phm cht tinh dch:
Trc khi cho n vin HASAMIN, Placebo.
Sau 10 ngy b sung vin HASAMIN, Placebo.
Sau 20 ngy b sung vin HASAMIN, Placebo.
Liu lng v cch cho n: Liu lung b sung: 01 vin/con/ngy. Cch b
sung vin thuc: trn u thuc vi mt t thc n tinh, cho th n ht, sau mi
cho n thc n c s.

74

Phn II. kt qu nghin cu

A. Thu thp mu v sng lc hot tnh sinh hc
Chng I. Thu thp, nh loi mu sinh vt bin v xy
dng c s d liu
I.1. tnh hnh nghin cu nhm Hi min, Da gai, San h mm
I.1.1. Tnh hnh nghin cu Hi min
Hi min (Porifera) l nhm sinh vt a bo nguyn thy c cu trc c
th kh n gin, c th tm thy ti hu ht cc thy vc, nhng ch yu bin.
Hin nay, c tnh trn th gii c khong 9000 loi. Hi min dinh dng bng
cch lc nc bin, v th n c kh nng lm sch nc bin thng qua cc hot
ng sng ca mnh. Trong qu trnh dinh dng Hi min cng thu nhn mt
s cht c c tnh, c tit ra bi mt s loi ng, thc vt (cc loi san h
sng trn rnvv). Thng qua cc qu trnh sinh hc phc tp, cc cht ny s
c hp thu v s dng nh l kho ha cht cho nhng mc ch khc nhau
ca chng (dng chng li cc loi ng vt n tht, ng vt k sinh v c ch
cc sinh vt khc trong vic cnh tranh ngun thc n v khng gian sng).
Hin nay Hi min ang l i tng c quan tm trong lnh vc tm
kim nhng ngun dc liu c ngun gc t nhin v thu c cc kt qu
tt trong vic tm ra cc hp cht chng ung th v mt s bnh khc. Mt s ni
nh New Zealand tin hnh nui trng mt s loi Hi min c gi tr phc
v cho mc ch thng mi cng nh cho vic nghin cu chit xut cc cht c
hot tnh sinh hc.
Tnh hnh nghin cu Hi min trn th gii
Hi min t lu thu ht c s ch ca nhiu nh khoa hc, vic
nghin cu khu h din ra t kh sm, c th chia qu trnh nghin cu thnh mt
s giai on sau
- Giai on 1: Cui th k 18 n gia th k 19
Mu vt thu thp mang tnh n l, ri rc v cc cng trnh cng b cn
ngho nn v khng r rng v iu kin t nhin ca khu vc nghin cu, in
hnh l cc cng trnh ca cc tc gi: Esper, 1797, 1830; Grant, 1836 trnh by
cc vn tng qut v gii ng vt Gray, 1858, 1870 m t mt s chi mi
trong lp Hi min vi (chi Aphroceras) ti qun o Malacca v mt s chi
ti vng ty Australia, Bowerbank & Norman, 1869, Bowerbank, 1872, 1877
trnh by nhng vn tng quan v Hi min v m t 5 loi mi qun o
Philippin v New Guinea.
- Giai on 2: Cui th k 19 n gia th k 20
Mu vt c thu thp trong cc cuc thm him ca cc nh khoa hc
Chu u, cc m t chi tit hn nh: Carter, 1883, 1887 nu v s phn b v
cc kin thc v Hi min, trong khong thi gian t 1873-1876 c cuc hnh
trnh vng quanh th gii ca tu Challenger, trn tu tp hp ng o cc nh
khoa hc lc by gi. Mu vt thu thp c trong chuyn hnh trnh ny cung

75

cp kh nhiu thng tin cho cc nh khoa hc v mt lot cc bo co khoa hc

c cng b sau hnh trnh ca tu Challenger in hnh l cc bo co sau:
Polesjaeff, 1884 cng b v thnh phn loi Hi min vi Calcarea thu thp
c trong chuyn hnh trnh, Ridley & Dendy, 1886, 1887 cng b v nhm Hi
min Monaxonida; Sollas, 1886, 1888 cng b v nhm Hi min
Tetractinellidae; ngoi ra cn c cc cng trnh cng b khc nh ca Lindgren,
1897, 1898 cng b v khu h Hi min ti qun o Malaysia, Topsent, 1897
cng b nhng nghin cu v Hi min ti vnh Amboine, Sollas, 1902 cng b
v Hi min ti vng bin Malaysia; Vosmaer, 1902 m t chi mi Placospongia
v Spirastrella ti vng bin Siboga, Wilson, 1925 cng b v nhm Hi min
silc v nhm Hi min song thu thp trong chuyn thm him ti qun o
Philippin, Ijima, 1926 cng b v thnh phn loi ca Hi min Hexactinellida
ti vng bin Siboga; Burton, 1928; Burton & Rao, 1932; Dendy & Burton, 1926;
bo co tnh hnh mu vt v Hi min sng ti cc vng nc nng v cc vng
nc su trong bo tng n , de Laubenfels, 1935 cng b v cc nghin cu
v Hi min thu thp t Puerto Galera Mindoro, Philippin.
- Giai on 3: T cui th k 20 n nay
C rt nhiu mu vt c thu thp nhng phn ln cc mu vt trn vn
cha nh loi chnh xc hoc cha nh loi c, phn ln cc nghin cu
chuyn sang nghin cu v cc c ch trao i cht ca Hi min, cc cng trnh
v nh loi cn li rt t tiu biu l: Caberoy, 1979, 1981; Caberoy & Tahil,
1983 cng b nghin cu v khu h Hi min ti Philippin (vnh Tayabas);
Esmero, 1978; cng b v thnh phn loi Hi min vng triu sng trn cc gi
th nhn to ti cng Cebu Philippin; Levis C., 1961 cng b nghin cu v Hi
min ti vng bin Nha Trang Vit Nam v vng bin Philippin; Pulitzer-Finali,
1980 nu mt s nghin cu v Hi min ti cc vng nc nng Hng Kng;
Van Soest, 1980, 1989, 1990 cng b nghin cu v khu h Hi min khu vc
Innxia. Hofman & Van Soest, 1995 m t cc loi thuc chi Lissodendroryx
(b Poecilosclerida, h Coelosphaeridae); Hooper et al., 1993 mt t loi
Oceanapia sagittaria (b Haplosclerida, h Phloeodictyidae); Hooper et al. 1994
cng b danh mc cc loi Hi min ca Australia. Gn y nht trong d n ko
di 7 nm tp hp gn 50 cc chuyn gia v phn loi Hi min do Hooper &
Soest ch tr cng b cun sch Systema Porifera y l h thng phn loi
mi nht c chnh sa.
Tnh hnh nghin cu hi min ti Vit Nam
Vit Nam t trc cho ti nay cha c cng trnh nghin cu no nghin
cu mt cch k lng khu h Hi min m ch yu vic nghin cu i tng
ny c kt hp vi vic nghin cu ngun li sinh vt. Cc cng trnh nghin
cu ch c cp ti trong cc bo co v thnh phn loi trong khu vc bin
ng hoc khu vc n Thi Bnh Dng ca mt s tc gi, cc bo co v
thnh phn loi Hi min in hnh c th lit k (theo th t thi gian) nh sau:
- Bo co ca Lindgren N.G., 1897, 1898 cng b khong 20 loi Hi min ti
khu vc qun o Malaysia v vng bin ng
- Lvi. C. 1961 cng b 28 loi Hi min ti khu vc bin Nha Trang

76

- Hooper J.N.A., Kennedy J.A. & van Soest R.W.M., 2000 cng b danh sch
hn 1500 loi Hi min thuc 102 h ca vng bin ng, trong c Vit Nam.
- Nm 2002 cc chuyn gia hot ng trong d n hp tc Vit Nam Italia v
bo tn a dng sinh hc di ven bin Vit Nam khi nghin cu ti khu vc vnh
H Long (Qung Ninh) v khu m ph Tam Giang Cu Hai (Tha Thin Hu)
pht hin 161 loi Hi min thuc 41 h.
I.1.2. Tnh hnh nghin cu da gai bin Vit Nam
Da gai (Echinodermata) c khong 350 loi thuc 58 h, 5 lp (Hu Bin,
Hi Sm, Sao Bin, Cu Gai v ui Rn) sng bin Vit Nam. Trong s
loi tp trung nhiu mt s h nh Sao bin (Astropectinidae), Holothuriidae
(Hi sm), Cucumariidae (Da chut bin), Amphiuridae (ui rn). Cc kt qu
nghin cu xc nh c khong 200 loi Da gai phn b vnh Bc B, 200
loi vng bin Trung B v 100 loi vng bin Nam B. Cc loi thng gp l
hi sm (Holothuria vagabunda), cu gai (Diadema setosum), sao bin
(Astropecten monocanthus, A. Polycanthus, A. velitaris), ui rn (Ophiactis
savygni) v.v.
Sinh vt lng ca nhm da gai tng i cao, chim n 29 % tng sinh
vt lng ng vt y vnh Bc B, 39 % vng bin Thun Hi - Minh Hi, 51
% vnh Vn Phong Nha Trang.
Da gai phn b nhiu habitat khc nhau, nhng thng tp trung cao
cc rn san h, rn hoc cc bi triu ct si + v sinh vt thuc cc Tng ng. Cc kt qu nghin cu v nhm ny cha nhiu, nhng xc nh mt s
qun x in hnh vnh Bc B m vai tr ch o thuc v nhm Da Gai. iu
ny c khng nh khi nghin cu chi tit 11 qun x sinh vt y phn b
vnh Bc B, ngi ta tm thy qun x s 2 phn b su 60 - 80 m
nc nm pha nam vnh, cc loi thuc nhm ui rn l nhm sinh vt ch
o trong qun x ny. Thm ch su 100 - 160 m, nm ra ngoi thm lc
a nhm Da Gai (Hu bin, ui rn) vn ng vai tr u th trong qun x.
Vn ng quan tm l trong hai nm 2003 - 2004 pht hin 19 loi c kh
nng cha cc dc liu quan trng. Thm ch c nhng loi khng ch cha
dc liu m cn pht trin mnh, to sinh khi ln c kh nng tr thnh cc bi
dc liu bin ln ca nc ta nh cc bi Sao bin Ct B, bi Hi Sm - Cu
Gai C T. Bi Vn Bi - Vn H cn gi l Bi Sao bin c din tch rng
khong 5 ha (50.000 m2), y l ni tp trung ca cc loi sao bin rnh nng
v sao bin rnh su (Astropecten polyacanthus, A. monacanthus) vi mt cao
(20 con/m2) bnh qun t n 0,3 kg/m2 (vo thng 3). Vi khi lng ny, ring
bi Vn Bi, Vn H c tr lng t 15 tn sao bin v cho kh nng khai thc
7,5 tn/nm. Ngoi sao bin cn c th khai thc cc loi Hi min cnh (Gellius
varius), hi sm en (Holothuria martensii), vnh (Isognomum isognomon), san
h mm (Sarcophyton sp), cu gai (Diadema setosum), hu ng (Ostrea
glomerata). Bi san h, cu gai, hi sm phn b ti khu vc t Bi Hng Vn
n hn Khe Tru (C T), mi ngy ngi dn a phng khai thc c t 20
- 50 kg hi sm en (Holothuria vagabunda) mang bn, bnh qun khong 30
kg/ngy, mt thng thu 900 - 1000 kg hi sm. Hi sm en thng sng trn cc

77

rn san h v theo nc triu tp vo cc bi ct kim n. Trn mi mt ct di 50

m, rng 3 m chng ti thu c t 20 - 30 con hi sm nh vy c khong 5 - 10
m2 tm thy 1 con hi sm sng. Ngoi hi sm y cn l ni tp trung ca cu
gai (Diadema setosum). Cu gai phn b vi mt cao trn cc rn san h ti
Hng Vn, thm ch c ni t n 5 - 10 con/m2. mt ny hon ton cho
php chng ta khai thc mt lng mu ln hng tn tin hnh chit xut cc
cht cn thit
I.1.3. Tnh hnh nghin cu san h mm trn th gii v Vit Nam
B San h mm (Alcyonacea) thuc lp ph San h tm tia Octocorallia,
lp San h Anthozoa, chng thng sng thnh tp on v phn b trn cc rn
san h vng bin Nhit i. Cng ging nh tn gi ca chng, San h mn c
nhiu tht v mm v chng khng c b khung xng cng bn trong nh san h
to rn. c im ni bt d nhn bit san h mm vi cc nhm san h khc l
cc polyp (c th) thng c 8 xc tu bao quanh ming c chc nng bt mi,
nhiu loi cn cha c t hoc cc t bo thch ty bo v. San h mm cn
gp phn vo s phong ph ca cc rn san h trong lng i dng, nhm ny
tuy khng c chc nng to rn nh nhm san h cng song chng cng l hp
phn quan trng to nn s phong ph v a dng cho rn san h v cng l ni
tr n ca nhiu loi sinh vt bin. Nhiu loi san h mm c hnh dng v mu
sc sc s cn c nui trong cc Aquarium lm cnh.
Vic nghin cu san h mm trn th gii cng mi ch c tin hnh t
khong na cui ca th k 20, c nhiu nhng cng b ca cc tc gi nh
Alderslade (1983, 1986, 2000), Benayahu (1984, 1991), Coll (1982), Williams
(1992, 1995, 1999) v nhiu tc gi khc. Tuy nhin cc tc gi mi gii quyt
c phn no v thnh phn loi v s phn b ca san h mm trn mt s rn
san h hoc m t mt ging loi no cha mang tnh chuyn su v ton din.
Nm 1984, mt tc gi ngi M Frederick Bayer cho xut bn cun San h
tm tia khu vc n Thi Bnh Dng y c coi l cun m t y nht
v b san h ny, song mi ch tp chung m t mt s loi ph bin. Sang u
th k XXI hai tc gi l Katharina Fabricius v Philip Alderslade thuc Vin
Khoa hc bin Australia xut bn cun San h mm v qut bin (Soft corals and
Sea fans). Cun ny m t y cc cc ging h ca b Alcyonacea, bao gm
90 ging thuc 23 h song cng ch dng li mc ging cha c m t n
loi. Ngoi ra cn c nhiu tc gi khc a ra cc kt qu nghin cu v th
nghim nui san h mm trong Aquarium lm cnh v sinh sn nhn to.
Vit nam vic nghin cu san h mm cn kh mi m, bt u t
nhng nm 90 ca th k trc tr li y v cha c nghin cu mang tnh
chuyn su. Cc tc gi nghin c v san h mm c th k n l V S Tun
(Vin Hi dng hc Nha Trang) v Nguyn Huy Yt (Phn vin Hi dng hc
Hi Phng), Nguyn ng Ngi (2004). Cc tc gi ny mi ch nu ln c
danh mc loi san h mm km theo danh mc san h cng trong cc bo co
iu tra mt s khu vc Ph Quc, Cn o, Trng Sa, C Lao Chm, Ct
B v cha c nhng cng b chnh thc hoc nghin cu chuyn su ring v
nhm san h ny. Tuy nhin theo c tnh vng bin Vit Nam c khong 130

78

loi san h mm, ring Ph Quc 19 loi. Gn y nht Vin Ho hp cht Thin
nhin kt hp vi Vin Ti nguyn v Mi trng Bin nghin cu v a ra
mt s kt qu v s chit xut cc hp cht trong 2 loi san h mm phn b ph
bin H Long - Ct B v Chn My. Theo kt qu nghin cu ban u hai loi
ny c th s dng lm dc liu cha mt s bnh nan y. y c coi l
nghin cu chuyn su v c tnh t ph trong lnh vc khai thc ti nguyn t
nhin trong lng bin lm thuc.
I.2. a im v thi gian thu mu
ti t chc 9 chuyn thu mu (Ct b thng 11/2006; Cn C thng
11/2006; Trng Sa thng 3-4/2007; Cn o thng 6/2007; Vnh H Long
thng 7/2007 ; Ct B thng 8/2007; Cn C t 6 n 10/6/2008; Ct B t 27/
6 n 5/7/2008; Vnh H Long t 9/9 n 16/9/2008). Tng s mu thu c
trong ti ny l 101 mu sinh vt Bin. C th bn cc t thu mu c
trnh by trn hnh di y:

Vnh H
Long
Ct B

Cn
C

Trng
Sa

Cn o

Cc a im thu mu nm 2006 - 2008.

79

I.3. Mt s nt c bn v c im mi trng cc a
im nghin cu
I.3.1. Khu vc Ct B
a im thu mu ch yu thuc Vnh Lan H nm pha ng o Ct B,
thuc huyn Ct Hi, thnh ph Hi Phng, c to a l 20043152004800 v Bc v 10700347- 10700755 kinh ng, cch cng Hi
Phng khong 60 km theo cc tuyn ng ni thu.
a hnh.
Vnh Lan H l khu vc vnh su trung bnh 13 mt v su nht t n 18
m, c nhiu ni vi che chn. Pha ng l dy ni Yn Nga thuc o Hang
Trai, pha ty l dy ni Rt Rt thuc o Ct B, pha bc l Ca Vn rng
khong 700 mt thng vi khu vc vnh H Long vi nhiu o vi sp xp
xen k to thnh mt bc tng chn sng kh l tng. Pha nam l phn ca
vnh, rng khong 5 km thng vi bin ng. Khu vc vnh c che chn c 3
mt ng, ty, bc bi cc o vi ln nh nn tng i kn sng, gi. Nm
trong vnh Lan H thuc khu vc o Ct B c nhiu tng vng nh nh: Tng
Ln Quay, Tng Gi, Tng Gu.
+ c im trm tch ca vnh:
- Bao gm cc dng trm tch ch yu l ct vi v sinh vt c cung cp t
cc sn san h, phn b ph bin ven b vnh Lan H, to nn cc bi ni ting
nh: bi Ty Tm, Ct Quyn, Ct Da, Ct Da con, Cng Quan, Vn Bi v.v.
Bi ct vi l thnh to rt c th, khng th gp c ni no khc ca Vit
Nam ngoi Ct B, Long Chu v vnh H Long - Bi T Long.
- Trm tch rn san h l mt dng tch t trm tch sinh vt, ch yu t san
h cht, Thn mm, Gip xc, Da gai, Hi min, Trng l v To vi. Trm tch
b ri trn mt rn ch yu l cc thnh phn v vi ht th nh ct t v ca
nhiu nhm sinh vt v gravel, thm ch l cui, tng v xc ca san h cnh v
dng khi. Thnh phn vt cht trm tch ch yu l carbonate, thng t trn
90%, phn cn li l mnh vn vi, vi silic. Dng trm tch ny phn b
cc tng, vng bao quanh vnh Lan H. Rn san h trong khu vc c quy m nh
thng khng qu 1ha. Theo kim k s b, tng din tch phn b cc rn san h
khu vc Lan H v ln cn (k c Hang Trai v u B) ch vo khong trn
70ha, thng phn b khong su 0,7 - 7,0m (Kho st Vit - Nga, 1997).
Rn san h va l dng tch t, va l ngun cung cp trm tch to nn cc bi
ct vi lin k. Trn mt cc rn san h cht thng c ph bi lp mng bn
st.
- Trm tch doi ct triu, delta triu. Trm tch ny thnh to do dng triu rt,
to nn cc dng tch t ngm, thng xut hin vng ca Lch Triu ln nh
lch Ca Vn, ca cc tng, ng (embayments and shelters) v Lch Huyn.
Thnh phn c hc ca trm tch gm ch yu l ct ln mnh vn v sinh vt v
gravel.
- Trm tch vng vnh. Trm tch ny phn b ph bin y vnh Lan H v
y cc tng, ng ln cn vnh. Thnh phn c hc gm ch yu l bn st v st
bt, mu xm v xm en, t cao. Bn st y vnh Lan H c hm lng

80

cp ht nh hn 0,05mm c th t ti 70 - 98%, thnh phn carbonate t hm

lng ti 70%. cp ht ln hn 0,05mm (bn bt), thnh phn thch anh trong
khong 7 - 30%, khong vt st chim 11 - 13%, v sinh vt chim 21 - 43%, rt
t mn hu c v c bit c c mnh vn than tuy hm lng khng ng k.
Thu vn
nh hng ca cc sng pha lc a tuy khng ln i vi khu vc vnh Lan
H ni chung, nhng v ma ma do khi lng l t cc sng vng Qung Ninh.
- Hi Phng kh ln nn c nhng nh hng nht nh ti vng nghin cu. Cn
phi xt n cc sng chy vo vnh H Long nh: Tri, M, Man, Din Vng...
chy qua ca Lc, Ht, Bnh Dng t pha ty bc. c im cc sng ny ngn,
rng lng sng nh nn sc cha khng ln. Vo ma ma (thng 5-9) lng
nc ma t thng lu dn vo cc sng v ra vnh H Long tng i ln,
nn lu lng v tc dng tng nhanh. Vo ma l lu lng nc sng tng
hng chc ln nn phn no nh hng n khu vc nghin cu, lm nht khi
lng ven b, to ra dng chy mt vi quy m nh. Ngoi ra cc sng Cm,
Bch ng cng gp phn ng k vo ch thu vn ca khu vc ny.
- Thu triu: khu vc nghin cu c tnh cht nht triu u, bin cc i
4,3 mt v chm pha hn Hn Du 20 - 30 pht. Mi thng c 2 k triu cng,
mi k 11 - 13 ngy xen k 2 k triu km, mi k 3 - 4 ngy. Trong mt chu k
ngy m, thi gian triu dng khong 12 gi, thi gian triu rt khong 13 gi.
V ma h, triu cao vo cc thng 5, 6, 7 v thp vo cc thng 8, 9. V ma
ng, triu cao vo cc thng 10, 11, 12 v thp vo cc thng 3, 4.
- Dng chy: khu vc nghin cu chu nh hng ca h thng hon lu vnh Bc
B v c c im ring ca vng bin kn ven b ty bc vnh. Do c im a
hnh v hnh thi ng b phc tp, nhiu lch nh v cc o, phn b su
khng u n lm cho h thng dng chy y bin i mnh. Dng chy
tng hp hnh thnh ch yu do dng triu, dng gi thay i theo. ma gi ng
bc dng chy hng ty nam t vn tc trung bnh 20 - 30 cm/s. Trong khu vc
vnh Lan H, dng chy hng ty bc t u B v t pha Ct Da lc triu ln,
nhp li ca Vn v hng ln pha bc. Tc dng triu ln khong 15 cm/s
ca Vn v 17cm/s khu vc gia vnh Lan H. Dng triu xung c hng
ngc li, khi bt u chy xung, xut hin dng chy xoy t u B qua trung
tm vnh v pha Ct Da.
Tc dng chy xung t 25 - 41 cm/s. lch hp nh ca Vn hay Ct
Da, Hn Bt... tc dng ln hn trong vnh. Trong t kho st thng 3/2002,
gi tr dng chy trong khu vc vnh Lan H khng c t bin v gi tr so vi
c trng chung ca vnh. Khu vc ca vnh t gi tr ln nht 24cm/s, khu vc
gia vnh t gi tr nh nht 3 cm/s . Gi tr trung bnh ca cc trm mt rng
t 12cm/s, hng dng chy thnh hnh hng bc khi triu ln v hng nam
v ng nam, ty nam khi triu xung.
c im thu ho v cht lng nc
+ Nhit nc
Nhit nc bin ca vnh Lan H gn lin vi phng nn chung v nhit
nc ca ton khu vc Ct B - vnh H Long v thay i kh ln, ph thuc vo

81

nhit kh quyn. Trong nm, nc bin c nhit thp nht vo thng 2

thng 3 v bin i khong 19,5oC - 33,5 0C v cao nht vo thng 7 nhit ca
lp nc mt c th t ti 33,5 oC (hnh 3).
+ c ca nc
Trong nm c ca nc bin Vnh Lan H thay i khng ln v c s
khc bit ln gia cc ma :
- Vo ma kh, c ca nc nm trong khong t 8 - 12FTU, trung bnh
l 10 FTU.
-Vo ma ma, c ca nc tng cao, giao ng t 10- 20 FTU, trung
bnh l 15 FTU.
Trong c hai ma, c ca lp nc y lun cao hn so vi lp nc mt.
+ mui
mui ca nc bin vnh Lan H dao ng t 17 0/00 n 340/00 (hnh 5)
- Vo ma kh, mui cao v n nh, nm trong khong t 31 - 340/00.
mui trung bnh khong 32,50/00. Trong thi gian ny c s phn tng mui
kh yu: mui ca nc tng y cao hn so vi nc tng mt cha ti 10/00,
duy nht khu vc Ca vn tng y > mt ti 2 0/00.
- Vo ma ma, mui thay i trong mt khong kh rng, t 170/00 n
300/00 do nh hng ca nc lc a, trung bnh l 280/00. Trong ma ny,
mui phn tng rt mnh , mui ca nc tng y cao hn so vi nc tng
mt t 1 n vi phn nghn, thm ch ti 11 0/00 ti Bn Bo.
+ pH
pH ca nc bin vnh Lan H bin i trong mt khong hp, t 7,8 n 8,3
+ Hm lng xy ho tan trong nc bin
Bin i trong khong t 5 - 7 mg/l. Mt s im c bit nh Bn Bo, Tng
Gu c th xung thp di 5 mg/L
I.3.2. Cn o
V tr a l
To a l ca Cn o 1060 32' - 106045' kinh ng v 8037' - 8048'
v Bc.
Vn quc gia Cn o l qun o thuc tnh B Ra - Vng Tu, nm
Bin ng Nam B, cch Vng Tu khong 200 km v pha Nam v thnh ph
Cn Th 110 km v pha ng. o trung tm l Cn Sn v c khong 14 hn
o nh xung quanh. Tng din tch ca vn gm 6000 ha trn cn v 34.500 ha
mt nc.
Kh hu
Cn o nm trong min kh hu nhit i - xch o, mt nm c hai ma gi:
Gi ma ng Bc v gi ma Ty Nam.
- Gi ma ng - Bc: Thi t thng 10 nm trc n thng 4 nm sau
vi tc trung bnh l 5 - 7 m/s, cc i 15 - 18 m/s. Hng gi thnh hnh l
hng ng - Bc v hng ng.
- Gi ma Ty - Nam: Tc trung bnh 5 - 7 m/s, cc i 20 - 25 m/s,
hng thnh hnh l Ty - Nam v hng Nam.
- Cc thng 4 v 9 l hai thng chuyn tip gia hai ma.

82

 Nhit :
- Nhit khng kh trung bnh nm 27,80C, giao ng trung bnh thng 26 0
29 C, bin giao ng khng qu 40C , thuc vo khu vc c nhit n ho.
- Nhit nc bin giao ng trong khong 25,7 - 29,20C, cao nht vo cc
thng 5 v thng 6, nhit giao ng 26,40C - 30,20C v thp nht vo cc thng
1 v 2 t 25,30C - 28,70C.
Ma v m
Lng ma trung bnh 2575 mm (N.V. Vit, 1984) v phn b 2 ma r
rt.
- Ma ma t thng 5 n thng 12 vi lng ma trung bnh t 200 - 250
mm/thng. Ma kh ngn hn, ch ko di 4 thng (t thng 1 - 4) lng ma
trung bnh ch t 35 mm/thng.
- m trung bnh 82 %, thp nht 60 %. Thuc o c m tt, thun li
cho thc vt pht trin.
Thu triu
Thu triu thuc vo loi bn nht triu khng u, cao cc i 4,1 m
v cc tiu 0,21 m, trung bnh 2,29 m.
Dng chy
Dng chy b chi phi bi hai yu t chnh l dng triu v gi ma. Vng
st o, dng chy ph hp vi dng triu ln v triu xung. Gi ma thng di
chuyn khi nc t pha Nam ln khi gi ma Ty - Nam v ph Bc xung vo
ma ng - Bc.
mn: mn trung bnh 31,9 %o v thp nht 15,4 %o, cao nht t
35 %o. Tuy nhin vo ma ma, do nc t trn o xung cng vi cc ca
sng ra, thng lm mn gim t ngt, gy nh hng n i sng thu
sinh vt khu vc ny.
I.3.3. Trng Sa
Khi Q Trng Sa gm ton b y bin v lng t rng ln ca phn
ng nam v nam Bin ng m cc o, cn ni v bi ngm ca Q Trng Sa
l trung tm. a khi Q Trng Sa bao gm cc kin trc a cht thng c
nhc n trong cc vn kin quc t vi cc tn gi: cc o Spratly, Dangerous
Grounds, Reed Bank, Luconia Shoals v Calimian Block hay bc ParawanMindoro (L Duy Bch v Ng Gia Thng, 2003).
Kh hu thi tit
Nm trong vng nhit i g ma in hnh ca ng Nam chu vi s
phn ho su sc theo ma kh hu. Ma gi Ty nam ko di t thng 5 n
thng 11, ma gi ng bc t thng 11 n thng 4 nm sau, trong thng 4
v thng 10 l 2 thng chuyn tip.
Nhit khng kh trung bnh gia cc ma dao ng khng ln, t 21,7
n 27,8OC
Lng ma nh hn trong t lin, trung bnh vo khong 1200 mm. S
gi nng cao, tm nhn xa ca cc ngy trong nm u t v vot trn 10 km v
khng c song m.

83

Gi thnh hnh trong ma gi ng bc l ng bc v ng, cn trong

ma gi Ty nam thnh hnh l ty nam v nam, cc loi gi khc chim tn sut
nh di 10%. Trong nm kh hu thi tit y thung chu nhiu nhiu lon:
p thp nhit i v bo. Theo thng k nhiu nm, s lng cc cn bo b
vo v i qua vng ln n 9-10 cn, chng thng gy sng ln vi cao cc
i c ghi nhn l 11m, hu hoi b bi v cc rn san h vng nc nng.
Ch hi vn
Ni chung, nhit nc tng mt Bin ng tng i cao, trung bnh
trn 23oC, trong ma h trn khp Bin ng l 29-30oC, bin ng theo ma v
theo v , nht l vo thi k gi ma ng bc. vng bin Q Trng Sa
cng mang c tnh , nhit nc trung bnh cao, 24-27oC, nhng s dao
ng theo ma khng qu ln vi bin l 3,4oC. Nhit nc bin thp nht
quan st c xung n 24,5oC.
mui ca nc tng i cao (trung bnh l 33,1) v kh n nh v
khng chu nh hng ca khi nc ra t lc a, tuy nhin, gi tr ny li c
xu hng bin ng theo ma vi bin 1,3 Song T ty v 1,7 khu
vc o Trng Sa, trong vo ma kh thng cao hn so vi ma ma (V
T Lp, 2003).
Theo chiu thng ng nc xut hin s phn tng r rt. Nhng kho st
lin hp Bin ng Vit Nam- Philippines-96 (L c T v Gil Jacinto, 1966)
cho thy, lp nc mt vi b dy khong 10-20 m kh ng nht, nhit v
mui t bin i (28-29 oC v 33,6-34,0). Tip theo l tng t bin nhit vi
gradient trung bnh 0,5 oC/m, cc i ln n 0,1- 0,2 oC/m ti cc su t 30-50
n 60-100 m, ty thuc vo a hnh y. Lp cui cng l lp thermocline vnh
cu, bt u t su 200-300 m, vi mui c gi tr cc i, trn 34 v
di lp ny nhit hu nh khng bin i na. Nhng nghin cu chi tit,
ring v dng chy khu vc Q Trng Sa rt t c cng b. ng nhin,
trong bc tranh chung, dng chy trong khu vc chu nh hng trc tip ca
hon lu Bin ng, lin quan n s xm nhp ca dng Korsivo qua eo bin
Luzon vo vng trong thi k gi ma ng bc. Nhn chung, vo ma h (thng
8) hng dng chy trung tm Bin ng theo chiu kim ng h, cn vo ma
ng (thng 12) dng chy c hng ngc li (Wyrtki, 1961).
Hm lng DO trong cc lp nc mt kh cao, t mc bo ho 95-105%
vi gi tr tuyt i khong 5,89-6,58 mg/l. Hm lng DO cc i thng nm
lp nc 25-50m.
Gi tr pH ca nc bin kh n nh, thng hi gh kim vi gi tr
trung bnh 8,09-8,28 (L Dc T v Gil Jacinto, 1996).
I.4. Kt qu thu mu
Trong ba nm 2006-2008, qua 9 t thu mu ti tin hnh thu thp
c 101 mu sinh vt Bin c kh nng cha dc liu ti cc a im nh sau:
I.4.1. Thu thp mu t 1 ti Ct B thng 11/2006
a. Thi gian, a im v s lng mu
- Thi gian thu mu: t ngy 5 n 15/11 nm 2006
- Cc im thu: Ct Da, Vn Bi, Ba Tri o, ...

84

- S mu thu: 6 mu.
b. Danh sch mu t 1 (11/2006)
Stt

Tn ting
Vit

Hi min
cnh xanh

Gellius varius

Hi min
xp
Hi min
Hi min
dy
Hi min
Hi min

Dysidea cinerea

3
4
5
6

Tn khoa hc

Haliclona sp
Mycale plumosa
Ircinia echinata
Amorphinopsis
excavans

a im thu mu

K hiu
thu mu

K hiu
ti

Vn Bi - Ct B

Sponge 1

KC-001

Vn Bi, Ba Tri o Ct B
Vn H-Ct B
Vn Bi, Ba Tri o Ct B
Vn Bi, Ba Tri o
Vn Bi, Ba Tri o

Sponge 2

KC-002

Sponge 3 KC-003
Spoinge 4 KC-004
Sponge 5
Sponge 6

KC-005
KC-006

c. Hnh nh mu t 1 (11/2006)
TT

K hiu

Thng tin v mu

KC-001

KC-002

KC-003

-Tn Vit nam: Hi

min cnh xanh
- Tn Khoa hc:
Gellius varius
- n v thu mu:
Vin Ti nguyn v
Mi trng Bin
- Ngi gim nh:
TS. Cng Thung
v cng s
-Tn Vit nam: Hi
min xp
- Tn Khoa hc:
Dysidea cinerea
- n v thu mu:
Vin Ti nguyn v
Mi trng Bin
- Ngi gim nh:
TS. Cng Thung
v cng s
-Tn Vit nam: Hi
min
- Tn Khoa hc:
Haliclona sp.
- n v thu mu:
Vin Ti nguyn v
Mi trng Bin
- Ngi gim nh:
TS. Cng Thung
v cng s

nh mu lu

a im, thi
gian, k hiu thu
mu
Vn Bi, Ct B.
Thng 11 nm 2006
K hiu mu thu:
Sponge 1

Vn Bi,Ba Tri o,
Ct B.
Thng 11 nm 2006
K hiu mu thu:
Sponge 2

Vn H, Ct B, Hi
Phng
Thng 11 nm 2006
K hiu mu thu:
Sponge 3

85

KC-004

KC-005

KC-006

-Tn Vit nam: Hi

min dy
- Tn Khoa hc:
Mycale plumosa
- n v thu mu:
Vin Ti nguyn v
Mi trng Bin
- Ngi gim nh:
TS. Cng Thung
v cng s
-Tn Vit nam: Hi
min
- Tn Khoa hc:
Ircinia echinata
- n v thu mu:
Vin Ti nguyn v
Mi trng Bin
- Ngi gim nh:
TS. Cng Thung
v cng s
-Tn Vit nam: Hi
min
- Tn Khoa hc:
Amorphinopsis
excavans
- n v thu mu:
Vin Ti nguyn v
Mi trng Bin
- Ngi gim nh:
TS. Cng Thung
v cng s

Vn Bi, Ba Tri o,
Ct B, Hi Phng
Thng 11 nm 2006
K hiu mu thu:
Sponge 4

Vn Bi, Ba Tri o,
Ct B, Hi Phng
Thng 11 nm 2006
K hiu mu thu:
Sponge 5

Vn Bi, Ba Tri o,
Ct B, Hi Phng
Thng 11 nm 2006
K hiu mu thu:
Sponge 6

I.4.2. Thu mu t 2, Cn C thng 11/2006

a. Thi gian, a im v s lng mu
- Thi gian thu mu: t ngy 5 - 20/11/2006
- Cc im thu: o Cn C - Qung Tr
- S mu thu: 9 mu
b. Danh sch mu t 2 (11/2006)
Stt

7
8

Tn ting
Tn khoa hc
Vit
San h mm Cladiella sp
San h mm Sarcophyton
trocheliophorum
San h mm Lobophytum
crassum
San h mm Lobophytum sp
San h mm Lobophytum
compactum
San h mm Lobophytum
compactum
Hi min
Amorphinopsis sp
San h mm Sinularia abrupta

San h mm Klyxum sp

1
2
3
4
5
6

a im thu mu
Cn C - Qung Tr
Cn C - Qung Tr

K hiu
thu mu
Cn c 1
Cn c 2

K hiu
ti
KC-007
KC-008

Cn C - Qung Tr

Cn c 3

KC-009

Cn C - Qung Tr
Cn C - Qung Tr

Cn c 4
Cn c 5

KC-010
KC-011

Cn C - Qung Tr

Cn C 6

KC-012

Cn C - Qung Tr
Cn C - Qung Tr

Cn C 7
Cn c 8

KC-013
KC-014

Cn C - Qung Tr

Cn c 9

KC-015

86

c. Hnh nh mu t 2 (11/2006)
Stt

Code
KC
KC-007

KC-008

KC-009

KC-010

KC-011

Thng tin v mu

nh mu lu

-Tn Vit nam: San h

mm
- Tn Khoa hc:
Cladiella sp.
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc:
Sarcophyton
trocheliophorum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc:
Lobophytum crassum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc:
Lobophytum sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc:
Lobophytum compactum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

a im, thi gian,

k hiu thu mu
Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c I

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c II

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c III

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c IV

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c V

87

KC-012

KC-013

KC-014

KC-015

-Tn Vit nam: San h

mm
- Tn Khoa hc:
Lobophytum compactum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi min
Tn
Khoa
hc:
Amorphinopsis sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c VI

-Tn Vit nam: San h

mm
- Tn Khoa hc:
Sinularia abrupta
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc: Klyxum
sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c 8

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c VII

Cn C, Qung Tr
Thng 11 nm 2006
K hiu thu mu:
Cn c 9

I.4.3. Thu mu t 3 Trng Sa thng 3-4/2007

a. Thi gian, a im v s lng mu
- Thi gian thu mu: t 20 thng 3 n 20 thng 4/2007
- Cc im thu: Trng Sa Ln, Nam Yt, Tc Tan
- S mu thu: 6 mu
b. Danh sch mu t 3 (thng 3 n thng 4/2007)
Stt

Tn ting Vit

Tn khoa hc

a im thu mu

Hi min

Petrosia
nigricans

Trng Sa Ln

Hi min hnh
nn
Hi min hnh

Ianthella sp

Nam Yt-Trng Sa

Xestospongia

Tc Tan - Trng Sa

88

K hiu
thu mu

K hiu
ti

Trng Sa
10

KC-016

Nam yt
11
Tc Tan

KC-017
KC-018

chung
Hi min hnh
chung
Hi min mng
mng
San h mm

4
5
6

testudinaria
Xestospongia
testudinaria
Ircinia sp
Lobophytum
compactum

Nam yt- Trng Sa

Tc tan - Trng Sa
Nam yt- Trng Sa

12
Nam Yt
13
Tc Tan
14
Nam Yt
15

KC-019
KC-020
KC-021

c. Hnh nh mu t 3 (thng 3 n thng 4/2007)

Stt

Code
KC

KC-016

-Tn Vit nam: Hi min

- Tn Khoa hc: Petrosia
nigricans
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

KC-017

-Tn Vit nam: Hi min

hnh nn
- Tn Khoa hc:
Ianthella sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

KC-018

-Tn Vit nam: Hi min

hnh chung
- Tn Khoa hc:
Xestospongia
testudinaria
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

KC-019

-Tn Vit nam: Hi min

hnh chung
- Tn Khoa hc:
Xestospongia
testudinaria
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Thng tin v mu

nh mu lu

a im, thi gian, k

hiu thu mu
Trng Sa ln, Khnh Ha
Thng 3-4 nm 2007
K hiu mu thu:
Trng Sa 10

Nam Yt, Trng Sa,

Khnh Ha
Thng 3-4 nm 2007
K hiu mu thu:
Nam Yt 11

Tc Tan, Trng Sa, Khnh

Ha
Thng 3-4 nm 2007
K hiu mu thu:
Tc Tan 12

Nam yt, Trng Sa, Khnh

Ha
Thng 3-4 nm 2007
K hiu mu thu:
Nam Yt 13

89

KC-020

-Tn Vit nam: Hi min

mng mng
- Tn Khoa hc: Ircinia
sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

KC-021

-Tn Vit nam: San h

mm
- Tn Khoa hc:
Lobophytum compactum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Tc Tan, Trng Sa, Khnh

Ha
Thng 3-4 nm 2007
K hiu mu thu:
Tc Tan 14

Nam yt, Trng Sa,

Khnh Ha
Thng 3-4 nm 2007
K hiu mu thu:
Nam Yt 15

I.4.4. Thu mu t 4 ti Cn o thng 6/2007

a. Thi gian, a im v s lng mu
- Thi gian thu mu: t 20 thng 5 n 10 thng 6/2007
- Cc im thu: Hn Tre, Hn By Cnh, Hn Trc, Vnh Cn Sn.
- S mu thu: 17 mu
b. Danh sch mu t 4 (thng 6/2007)
Stt

Tn ting Vit

Tn khoa hc

a im thu
mu

K hiu
thu mu

K hiu
ti

L vi 2

KC-022

Hi min cy

Clathria vulpina

Mi L Vi Cn o

San h mm

Sarcophyton elegans

am tre (Hn Damtre 3

Tre)

KC-023

San h mm

Lobophytum compactum

Hn By
Cnh - Cn
o

By cnh
04

KC-024

San h mm

Sarcophyton mililatensis

Hn By
Cnh - Cn
o

By cnh
05

KC-025

San h mm

Lobophytum compactum

Hn By
Cnh - Cn
o

By cnh
06

KC-026

Hi min

Petrosia nigricans

Hn By
Cnh - Cn
o

By cnh
07

KC-027

90

Hi min hnh
chung

Xestospongia
testudinaria

Hn By
Cnh - Cn
o

By cnh
08

KC-028

Hi sm en Cn
o

Holothuria atra

Hn By
Cnh - Cn
o

By cnh
09

KC-029

Hi min hnh qu Niphates sp

u

Hn By
Cnh - Cn
o

By cnh
10

KC-030

10

ui rn

Macrophiothrix sp

Hn By
Cnh - Cn
o

By cnh
11

KC-031

11

Hi min xp l

Spongia sp

Hn Trc Cn o

Hn Trc KC-032
12

12

San h mm

Sinularia sp

Hn Trc Cn o

Hn trc
13

KC-033

13

Hi min cnh

Clathria vulpina

Hn Trc Cn o

Hn trc
14

KC-034

14

Hi min phin Stylissa flabelliformis

Hn Trc Cn o

Hn trc
15

KC-035

15

San h mm hnh
chm chm

Dendronephthya
klunzeringi

Hn Trc Cn o

Hn trc
16

KC-036

16

Hi min

Gellius var fibrosa

Vnh Cn
Sn-Cn o

Cn Sn
17

KC-037

17

Cu gai hnh
trng

Echinocardium
cordatum

Vnh Cn sn Cn Sn
- Cn o
18

KC-038

c. Hnh nh mu t 4 (thng 6/2007)

Stt

Code KC

Thng tin v mu

KC-022

-Tn Vit nam: Hi

min cy
- Tn Khoa hc:
Clathria vulpina
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

nh mu lu

91

a im, thi gian, k

hiu thu mu
L vi, Cn o, B Ra
- Vng Tu
20 thng 5 n 10 thng
6/2007
K hiu mu thu: Lvi
2

KC-023

KC-024

KC-025

KC-026

KC-027

KC-028

-Tn Vit nam: San h

mm
- Tn Khoa hc:
Sarcophyton elegans
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc:
Lobophytum compactum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm hnh l gan
- Tn Khoa hc:
Sarcophyton mililatensis
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm
- Tn Khoa hc:
Lobophytum compactum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi
min
- Tn Khoa hc:
Petrosia nigricans
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi
min hnh chung
- Tn Khoa hc:
Xestospongia
testudinaria
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung &cs

Dam tre, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Damtre3

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh04

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh05

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh06

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh07

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh08

92

KC-029

KC-030

10

KC-031

11

KC-032

12

KC-033

13

KC-034

-Tn Vit nam: Hi sm

en Cn o
- Tn Khoa hc:
Holothuria atra
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi
min hnh qu u
- Tn Khoa hc:
Niphates sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Da gai
ui rn
- Tn Khoa hc:
Macrophiothrix sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi
min xp l
- Tn Khoa hc: Spongia
sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm hnh chn vt
- Tn Khoa hc:
Sinularia sp
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi
min cnh
- Tn Khoa hc:
Clathria vulpina
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh09

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh10

By cnh, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh11

Hn trc, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Bycnh12

Hn trc, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu: Hn
trc13

Hn trc, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu: Hn
trc14

93

14

KC-035

15

KC-036

16

KC-037

17

KC-038

-Tn Vit nam: Hi

min phin
- Tn Khoa hc: Stylissa
flabelliformis
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: San h
mm hnh chm chm
- Tn Khoa hc:
Dendronephthya
klunzeringi
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi
min cnh
- Tn Khoa hc: Gellius
var fibrosa
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam:
Da gai hnh trng
- Tn Khoa hc:
Echinocardium
cordatum
- n v thu mu: Vin
Ti nguyn v Mi
trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Hn trc, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu: Hn
trc15

Hn trc, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu: Hn
trc16

Con son, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Conson17

Con son, Cn o, B
Ra - Vng Tu
T 20 thng 5 n 10
thng 6/2007
K hiu mu thu:
Conson18

I.4.5. Thu mu t 5 ti Vnh H Long thng 7/2007

a. Thi gian, a im v s lng mu
- Thi gian thu mu: T ngy 3 - 15 thng 7/2007
- Cc im thu: u B, Hang Trai, Hang D (Ang D), Cng
- S mu thu: 16 mu
b. Danh sch mu t 5 (3 n ngy 15 thng 7/2007)
Stt

Tn ting Vit

Tn khoa hc

San h mm

Sarcophyton sp

San h mm

Lobophytum sp

San h mm

Lobophytum sp

a im thu
mu
Cng , H
Long
nh D, H
Long
nh D, H L

94

K hiu thu
mu
Cong Do 20

K hiu
ti
KC-039

Ang Du 21

KC-040

Ang Du 22

KC-041

Hi min hnh Haliclona

chung
subarmigera
C gim gai di
Diadema
setosum
Hi min
Ircinia echinata

Hi min xanh

8
9
10

Hi min hnh
ti
Hi min
Sao bin

11

Hi min

12
13

Hi min bt
Hi min tm

14

Sao bin 5 cnh

mu hng
Sao bin 5 cnh
mu tm
Sao bin 5 cnh
mu tm ru

15
16

Biemna cf.
Nudelringna
Xestospongea
testudinaria
Gelliodes fibulata
Anthenea
pentagonala
Aka mucosa
Dysidea cinerea
Haliclona
subarmigera
Protoreaster
nodusus
Asterope
carinifera
Asterope
carinifera

nh D, H
Long
B Xm, H
Long
nh D, H
Long
nh D, H
Long
B Xm, H
Long
B Xm, H L
Vn H, H
Long
B Xm, H
Long
nh D, H L
B Xm, H
Long
Vn H, H
Long
Vn H, H
Long
Vn H, H
Long

Ang Du 23

KC-042

H.63

KC-043

Hinh52

KC-044

Hinh54

KC-045

H.55

KC-046

Hinh57
Hinh60
(HL44)
Hinh61
(BX3)
Hinh99
BX4
(Hinh174)
VH3
(Hinh384)
VH2
(Hinh385)
VH1
(Hinh386)

KC-047
KC-048
KC-049
KC-050
KC-052
KC-053
KC-054
KC-055

c. Hnh nh mu t 5 (3 n ngy 15 thng 7/2007)

Stt

Code KC

KC-039

KC-040

KC-041

Thng tin v mu

nh mu lu

-Tn Vit nam:

San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam:
San h mm
Tn
Khoa
hc:
Lobophytum sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam:
San h mm
Tn
Khoa
hc:
Lobophytum sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

a im, thi gian,

k hiu thu mu
Cong Do, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Congdo 20

Ang Du, H Long,

Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Angdu21

ng Du, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Angdu22

95

KC-042

-Tn Vit nam: Hi min

hnh chung
- Tn Khoa hc: Haliclona
subarmigera
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam:
C gim gai di
- Tn Khoa hc: Diadema
setosum
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

ng Du, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Angdu23

KC-043

KC-044

-Tn Vit nam: Hi min

- Tn Khoa hc: Ircinia
echinata
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

ng D, H. Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
H.52

KC-045

ng D, H. Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh54

KC-046

-Tn Vit nam: Hi min

xanh
- Tn Khoa hc: Biemna cf.
nudelringna
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: Hi min
hnh ti
Tn
Khoa
hc:
Xestospongea testudinaria
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

KC-047

-Tn Vit nam: Hi min

- Tn Khoa hc: Gelliodes
fibulata
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

B Xm, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh57

10

KC-048

-Tn Vit nam:

Sao bin
- Tn Khoa hc: Anthenea
pentagonala
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Vn H, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh60

B Xm, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
H.63

B Xm, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
H.55

96

11

KC-049

12

KC-050

13

KC-052

14

KC-053

15

KC-054

16

KC-055

-Tn Vit nam: Hi min

- Tn Khoa hc: Aka
mucosa
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi min
bt
- Tn Khoa hc: Dysidea
cinerea
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi min
tm
- Tn Khoa hc: Haliclona
submarginera
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam:
Sao bin nm cnh mu
hng
Tn
Khoa
hc:
Protoreaster nodusus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam:
Sao bin 5 cnh mu tm
- Tn Khoa hc: Asterope
carinifera
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v s

B Xm, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh61

-Tn Vit nam:

Sao bin 5 cnh mu tm
ru
- Tn Khoa hc: Asterope
carinifera
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

Vn H, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh386

ng D H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh99

B Xm H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh174 (BX4)
Vn H, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh384

Vn H, H Long,
Qung Ninh
T ngy 3 n ngy
15 thng 7/2007
K hiu mu thu :
Hinh385

I.4.6. Thu mu t 6 ti Ct B thng 8/2007

a. Thi gian, a im v s lng mu
- Thi gian thu mu: T ngy 1 - 10 thng 8/2007
- Cc im thu: Ct Da, Vn Bi, Ba Tri o, ...
- S mu thu: 17 mu
97

b. Danh sch mu t 6 (T ngy 1 - 10 thng 8/2007)

Stt

Tn ting Vit

Tn khoa hc

a im thu
mu
Ca ng
Ti, Ct B
Tng Gu,
Ct B
Vn Bi, Ct
B
Vn Bi, Ct
B
Vn Bi, Ct
B
Ca ng
Ti, Ct B
Vn Bi, Ct
B
Ca ng
Ti, Ct B
Ca ng
Ti, Ct B
Ca ng
Ti, Ct B
Tng Gu,
Ct B

K hiu
thu mu
CB 17

K hiu
ti
KC-051

CB 4
(hinh23)
CB 1
(hinh215)
CB 3
(hinh260)
CB 6
(hinh262)
CB 7
(hinh303)
CB 2

KC-056

CB 8
(hinh309)
CB 10
(hinh315)
CB 16
(hinh316)
CB 9
(hinh327)

KC-062

Hi min trng

Mycale plumosa

Sao bin rnh nng

Da chut bin

Astropecten
polyacanthus
Holothuria spinifera

Hi min xp

Dysidea cinerea

Hi min xp

Biemna megalosigma

Hi min

Ircinia echinata

Hi sm

Holothuria scabra

Hi min hnh chp

Petrosia nigricans

Hi min hnh chp

10

Hi min

Spongia sp

11

San h mm c gai

Cladiella sp

12

San h mm

Sarcophyton sp

Tng Gu,
Ct B

CB 14
KC-066
(hinh361)

13

Sao bin 5 cnh mu

tm ru
Da chut bin mu
hng c gai
Da chut bin mu
hng khng gai
Sao bin cnh ngn
mnh dy
Sao bin

Asterope sp

Ca ng
Ti, Ct B
Vn Bi, Ct
B
Vn Bi, Ct
B
Ca ng
Ti, Ct B
Ca ng
Ti, Ct B

CB 13
(hinh387)
CB 11
(hinh388)
CB 5
(hinh390)
CB 15
(hinh391)
CB 12

14
15
16
17

Petrosia nigricans

Stichopus sp
Thyone bicornis
Asterina batheri
Archaster typicus

KC-057
KC-058
KC-059
KC-060
KC-061

KC-063
KC-064
KC-065

KC-067
KC-068
KC-069
KC-070
KC-071

c. Hnh nh mu t 6 (T ngy 1 - 10 thng 8/2007)

Stt

Code KC

KC-051

Thng tin v mu

nh mu lu

-Tn Vit nam: Hi min

trng
- Tn Khoa hc: Mycale
plumosa
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

98

a im, thi gian,

k hiu thu mu
Ca ng Ti, Ct B
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu :
Hinh100

KC-056

-Tn Vit nam:

Sao bin rnh nng
- Tn Khoa hc: Astropecten
polyacanthus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: Da chut
bin
- Tn Khoa hc: Holothuria
spinifera
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: Hi min xp
- Tn Khoa hc: Dysidea
cinerea
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

Tng Gu, Ct B,
Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu :
Hinh23

KC-057

KC-058

KC-059

-Tn Vit nam: Hi min xp

- Tn Khoa hc: Biemna
megalosigma
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam: Hi min
- Tn Khoa hc: Ircinia
echinata
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

Vn Bi, Ct B, Hi
Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu :
Hinh262

KC-060

KC-061

-Tn Vit nam:

Hi sm
- Tn Khoa hc: Holothuria
scabra
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

Vn Bi, Ct B, Hi
Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
CB2

KC-062

-Tn Vit nam: Hi min

hnh chp
- Tn Khoa hc: Petrosia
nigricans
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

Ca ng Ti, Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 309

Vn Bi , Ct B,
Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu :
Hinh215
Vn Bi, Ct B, Hi
Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu :
Hinh260

Ca ng Ti, Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu :
Hinh303

99

KC-063

-Tn Vit nam: Hi min

hnh chp
- Tn Khoa hc: Petrosia
nigricans
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

Ca ng Ti , Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 315

10

KC-064

-Tn Vit nam:

Hi min
- Tn Khoa hc: Spongea sp.
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

Ca ng Ti , Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 316

11

KC-065

-Tn Vit nam:

San h mm
- Tn Khoa hc: Cladiella
sp.
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

Tng Gu Ct B,
Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 327

12

KC-066

Tng Gu Ct B,
Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 361

13

KC-067

-Tn Vit nam:

San h mm
- Tn Khoa hc: Sarcophyton
sp.
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam:
Sao bin 5 cnh mu tm ru
- Tn Khoa hc: Asterope
sp.
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

14

KC-068

-Tn Vit nam:

Da chut bin mu hng c
gai
- Tn Khoa hc: Stichopus
sp.
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

Vn Bi, Ct B, Hi
Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 388

Ca ng Ti, Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 387

100

15

KC-069

16

KC-070

17

KC-071

-Tn Vit nam:

Da chut bin mu hng
khng gai
- Tn Khoa hc: Thyone
bicornis
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam:
Sao bin cnh ngn mnh dy
mu hng m
- Tn Khoa hc: Asterina
batheri
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam:
Sao bin
- Tn Khoa hc: Archaster
typicus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

Vn Bi, Ct B, Hi
Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 390

Ca ng Ti, Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
Hinh 391

Ca ng Ti, Ct
B, Hi Phng
T ngy ngy 1 - 10
thng 8/2007
K hiu mu thu:
CB12

I.4.7. Thu mu t 7 ti Cn C t 6 n 10/6/2008

a. Thi gian, a im v s lng mu
- Thi gian thu mu: T ngy 6 - 10 thng 6/2008
- Cc im thu: Bc Cn C
- S mu thu: 9 mu
b. Danh sch mu t 7 (6 n 10 thng 6/2008)
Stt

Tn ting Vit

Tn khoa hc

a im thu
mu

K hiu
thu mu

K hiu
ti

San h mm

Sarcophyton sp

Bc Cn C

C08-1

KC-072

San h mm

Sarcophyton sp

Bc Cn C

C08-2

KC-073

San h mm

Sarcophyton sp

Bc Cn C

C08-3

KC-074

San h mm

Sarcophyton sp

Bc Cn C

C08-4

KC-075

San h mm

Sarcophyton sp

Bc Cn C

C08-5

KC-076

San h mm

Sarcophyton sp

Bc Cn C

C08-6

KC-077

San h mm

Sarcophyton sp

Bc Cn C

C08-7

KC-078

San h mm

Sarcophyton sp

Bc Cn C

C08-8

KC-079

San h mm

Sarcophyton sp

Bc Cn C

C08-9

KC-080

101

c. Hnh nh mu t 7(6 n 10 thng 6/2008)

Stt

Code
KC

Thng tin v mu

KC-72

KC-73

KC-74

KC-75

KC-76

KC-77

-Tn Vit nam: San h mm

San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cng
s

nh mu lu

a im, thi
gian, k hiu
thu mu
Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-1

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-2

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-3

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-4

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-5

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-6

102

KC-78

KC-79

KC-80

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-7

-Tn Vit nam: San h mm

San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: San h mm
San h mm
Tn
Khoa
hc:
Sarcophyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v cs

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-8

Bc Cn C
T ngy 6 - 10
thng 6/2008
K hiu mu
C08-9

I.4.8. Thu mu t 8 ti Ct B t 27/ 6 n 5/7/2008

a. Thi gian, a im v s lng mu
- Thi gian thu mu: T ngy 27/6 5/7/2008
- Cc im thu: Vn Bi, Ba Tri o, Hang Trai, u B,..
- S mu thu: 7 mu
b. Danh sch mu t 8 (ngy 27/6 n 5/7/2008)
Stt
1
2
3
4
5
6
7

Tn ting
Vit
Hi sm
en
Sao bin
Sao bin
Cu gai
Cu gai
C ghim
Sao bin

Tn khoa hc
Holothuria vagabunda

Vn Bi, Ct B

K hiu
thu mu
CB08-11

Astropecten polycanthus
A. polycanthus
Echinothrix calamaris

Vn Bi, Ct B
Ba Tri o, Ct B
Ba Tri o, Ct B
Hang Trai, Ct B
u B, Ct B
Vn H, Ct B

CB08-15
CB08-17
CB08-18
CB08-22
CB08-25
CB08-29

Echinothrix calamaris
Diadema setosum
Astropecten
polycanthus

a im thu mu

K hiu
ti
KC-081
KC-082
KC-083
KC-084
KC-085
KC-086
KC-087

c. Hnh nh mu t 8 (ngy 27/6 n 5/7/2008)

Stt

Code
KC

Thng tin v mu

KC-81

-Tn Vit nam: Hi sm

en
Tn
Khoa
hc:
Holothuria vagabunda
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs

nh mu lu

a im, thi
gian, k hiu
Vn Bi Ct B
ngy
27/6
n
5/7/2008
K hiu mu:

CB08-11

103

KC-82

KC-83

KC-84

KC-85

KC-86

KC-87

-Tn Vit nam: Sao bin

Tn
Khoa
hc:
Astropecten polycanthus
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs

Vn Bi Ct B
ngy
27/6
n
5/7/2008
K hiu mu:

CB08-15

-Tn Vit nam:

Sao bin
Tn
Khoa
hc:
Astropecten polycanthus
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: Cu gai
Tn
Khoa
hc:
Echinothrix calamaris
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs

Vn Bi Ct B
ngy
27/6
n
5/7/2008
K hiu mu:
CB08-17

Ba Tri o Ct B
ngy
27/6
n
5/7/2008
K hiu mu:
CB08-18

-Tn Vit nam:

Cu gai
Tn
Khoa
hc:
Echinothrix calamaris
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam:
C ghim
- Tn Khoa hc: Diadema
setosum
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs
-Tn Vit nam: Sao bin
Tn
Khoa
hc:
Astropecten polycanthus
- n v thu mu: Vin Ti
nguyn v Mi trng
Bin
- Ngi gim nh:
TS. Cng Thung v cs

Hang Trai, Ct B
ngy
27/6
n
5/7/2008
K hiu mu:
CB08-22

u B, Ct B
ngy
27/6
n
5/7/2008
K hiu mu:
CB08-25

Vn H, Ct B
ngy
27/6
n
5/7/2008
K hiu mu:
CB08-29

I.4.9. Thu mu t 9 ti Vnh H Long t 9/9 n 16/9/2008

a. Thi gian, a im v s lng mu
- Thi gian thu mu: T ngy 9/9 16/9/2008
- Cc im thu: Vn H, ng D, Cng , B Xm, Cc 8, Cc 6
- S mu thu: 14 mu
104

b. Danh sch mu t 9 (ngy 9/9 n 16/9/2008)

STT
1
2
3
4
5
6
7
8
9
10
11
12
13
14

Tn VN
San h mm
Hi sm en
Hi min cnh
xanh
Sao bin

Tn khoa hc
Sarcaphyton sp
Holothuria
vagabunda
Gellius varius

Astropecten
polyacanthus
Sao bin
Astropecten
monacanthus
Hi min
Biemna cf.
nudelringna
Hi min hnh ti Xestospongea
testudinaria
Hi min cnh
Gelliodes
fibulata
Hi min xanh
Haliclona
clathrata
Hi min trng
Mycale
plumosa
Sao bin
Astropecten
polycanthus
Sao bin
Astropecten
monocanthus
San h mm
Lobophytum sp
Sao bin
Astropecten
polycanthus

a im
Vn H, HL
Vn H, HL

K hiu mu
HL01-31
HL02-32

Cod KC
KC-088
KC-089

ng D, HL

HL03-33

KC-090

Cng , HL

HL04-34

KC-091

Cng , HL

HL05-35

KC-092

B Xm, HL

HL06-36

KC-093

B Xm, HL

HL07-37

KC-094

B Xm, HL

HL08-38

KC-095

Cc 8, HL

HL09-39

KC-096

Cc 6, HL

HL10-42

KC-097

Cng HL

HL09-40

KC-098

Cng HL

HL09-44

KC-099

Cng , HL
Cc 6 HL

HL09-41
HL09-43

KC-100
KC-101

c. Hnh nh mu t 9 (ngy 9/9 n 16/9/2008)

Stt

Code
KC

Thng tin v mu

KC-88

-Tn Vit nam:

San h mm
Tn
Khoa
hc:
Sarcaphyton sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Vn H, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL01-31

KC-89

-Tn Vit nam: Hi sm

en
Tn
Khoa
hc:
Holothuria vagabunda
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Vn H, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL02-32

nh mu lu

105

a im, thi
gian, k hiu
thu mu

KC-90

KC-91

KC-92

KC-93

KC-94

KC-95

-Tn Vit nam: Hi min

cnhxanh
- Tn Khoa hc: Gellius
varius
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam:
Sao bin
Tn
Khoa
hc:
Astropecten polyacanthus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Sao bin
Tn
Khoa
hc:
Astropecten monacanthus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

ng D, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL03-33

Cng , H
Long
T ngy 9/9
16/9/2008
K hiu mu:
HL04-34

Cng , H
Long
T ngy 9/9
16/9/2008
K hiu mu:
HL05-35

B Xm, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL06-36

-Tn Vit nam:

Hi min
- Tn Khoa hc: Biemna cf.
nudelringna
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi min
hnh ti
Tn
Khoa
hc:
Xestospongea testudinaria
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi min
cnh
- Tn Khoa hc: Gelliodes
fibulata
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

B Xm, H Long
Thng 9 nm
2008
K hiu mu:
HL07-37

B Xm, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL08-38

106

KC-96

10

KC-97

11

KC-98

12

KC-99

13

KC-100

14

KC-101

-Tn Vit nam: Hi min

xanh
- Tn Khoa hc: Haliclona
clathrata
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam: Hi min
trng
- Tn Khoa hc: Mycale
plumosa
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam:
Sao bin
Tn
Khoa
hc:
Astropecten polyacanthus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Cc 8, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL09-39

-Tn Vit nam:

Sao bin
Tn
Khoa
hc:
Astropecten monacanthus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam:
San h mm
Tn
Khoa
hc:
Lobophytum sp
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s
-Tn Vit nam:
Sao bin rnh nng
Tn
Khoa
hc:
Astropecten polyacanthus
- n v thu mu: Vin Ti
nguyn v Mi trng Bin
- Ngi gim nh:
TS. Cng Thung v
cng s

Cng , H
Long
T ngy 9/9
16/9/2008
K hiu mu:
HL9-44

Cc 6, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL10-42

Cng , H
Long
T ngy 9/9
16/9/2008
K hiu mu:
HL9-40

Cng , H
Long
T ngy 9/9
16/9/2008
K hiu mu:
HL9-41

Cc 6, H Long
T ngy 9/9
16/9/2008
K hiu mu:
HL9-43

Trong s 101 mu thu c c 41 mu hi min, chim 40,6 % tng s

mu thu c, 31 mu san h mm (30,7 %) v 29 mu da gai (28,7 %). Trong
s :

107

 Hi Min
+ S lng: 41 mu
+ V d v s phn b cc mu hi min ti cc im thu mu:
- Cc kt qu phn tch ca chng ti xc nh c 6 mu ti khu vc
ng nam o Ct B (Gellius varius, Dysidea cinerea, Haliclona sp, Mycale
plumosa, Ircinia echinata, Amorphinopsis excavans)
- 4 mu cng tm thy o Trng Sa (Petrosia nigricans, Janthella sp,
Xestospongia testudinaria, Ircinia sp)
- 7 mu Hi min ti vng bin Cn o (Clathria vulpina, Petrosia
nigricans, Xestospongia testudinaria, Niphates sp, Spongia sp, Stylissa
flabelliformis, Gellius var fibrosa)
- Ti Vnh H Long cng thu c 7 mu (Haliclona subarmigera,
Dysidea cinerea, Ircinia echinata, Biemna cf. nudelringna, Xestospongea
testudinaria, Gelliodes fibulata, Aka mucosa)
San h mm
+ S lng: 31 mu
+ V d v s phn b cc mu san h mm ti cc im thu mu:
- Ti Cn C tm thy 7 mu: Cladiella sp,
Sarcophyton
trocheliophorum, Lobophytum crassum, Lobophytum sp, Lobophytum
compactum, Sinularia abrupta, Klyxum sp
- Ti o Nam Yt thuc Trng Sa thu c 1 mu san h Lobophytum
compactum
- Ti Cn o thu c 5 mu: Sarcophyton elegans, Sarcophyton
mililatensis, Lobophytum compactum, Sinularia sp, Dendronephthya klunzeringi
- Ti H Long mi ch thu c 3 mu nm trong hai ging Sarcophyton v
ging Cladiella
- Ti Ct b thu c 3 mu thuc hai ging Sarcophyton v ging
Cladiella.
Da Gai
+ S lng: 29 mu
+ V d v s phn b cc mu da gai ti cc im thu mu:
- Ti Cn o: thu c 3 mu gm: Hi sm en (Holothuria atra),
ui rn (Macrophiothrix sp), cu gai hnh trng (Echinocardium cordatum)
- Vnh H Long: S lng mu thu c nhiu hn khu vc Cn o, tng
s 5 mu gm: Sao bin 5 cnh mu tm ru (Asterope carinifera), sao bin cnh
ngn (Anthenea pentagonala), sao bin 5 cnh mu tm (Asterope carinifera),
sao bin 5 cnh mu hng (Protoreaster nodusus), c gim gai di (Diadema
setosum)
- Vng bin Ct B : Trong t thu mu thng 8/2007 thu c 6 mu
Chng bao gm hi sm gai (Holothuria spinifera), sao bin rnh nng
(Astropecten polyacanthus), da chut bin mu hng khng gai (Thyone
bicornis), da chut bin mu hng c gai (Stichopus sp), sao bin 5 cnh mu
tm ru (Asterope sp) v sao bin cnh ngn.

108

I.5. M t cc loi sinh vt bin in hnh c kh nng cha

dc liu
I.5.1. Cc loi san h mm
1. Loi Lobophytum compactum, thuc h Alcyoniidae
c im: Tp on c dng ph dy,
trn polyparium c cc thu mc gn
nhau, di 3-5cm hu nh khng phn
nhnh. Polyp c b mt trn ca tp
on v trn cc thu
Mu sc: c mu nu nht, xanh nu, pht
hng

2. Loi Lobophytum crassum, thuc h Alcyoniidae

c im: Tp on c thn ngn v to
hoc dng ph. Polyparium phn nhnh
thnh cc thu hoc dng mo, rt him
khi c dng phng. Polyp lng hnh. Gai
xng trn b mt c dng di cui, gai
xng trong dng hnh con sut
Mu sc: c mu vng sng, nu hoc
xanh lc.

3. Loi Sarcophyton elegans, thuc h Alcyoniidae

c im: Tp on c mt thn ln, pha
trn dng a mang polyp gi l
polyparium. gia polyparium c dng
lng cho, pha ngoi gn sng hoc gp
khc. mi trng sng mnh thn ngn
v mp, mi trng yn tnh thn di v
mnh hn
Mu sc: c mu nu sm, vng nu hoc
xanh nu

109

4. Loi Sarcophyton mililatensis, thuc h Alcyoniidae

c im:
Tp on c thn ln,
polyparium c dng lng cho, pha ngoi
gn sng hoc gp khc. B mt tp on
nhm, polyp nh. mi trng sng mnh
thn ngn v mp, mi trng c bo
v thn di v mnh hn
Mu sc: c mu vng xanh, nu hoc nu
vng
5. Loi Sarcophyton trocheliophorum, thuc h Alcyoniidae
c im: Tp on c dng nm, thn
ln. B mt polyparium thng gp khc.
Polyp lng hnh. Gai xng trong c
hnh dng con sut vi cc mu li, gai
xng ngoi hnh di cui
Mu sc: c mu vng, nu nht hoc
xanh nht
6. Loi Sinularia abrupta, thuc h Alcyoniidae
c im: Tp on dng ph hoc c
thn phn bit. Polyparium nhn, phn
thu dng ngn hoc dng mo. Polyp
n hnh. Gai xng trong c dng con
sut ln c mu nh, chiu di ln hn
2mm. Gai xng ngoi dng di cui hoc
con sut nh.
Mu sc: hi trng, vng nht, nu sng

I.5.2. Cc loi hi min

1. Loi Gellius varius, thuc h Chalinidae
M t: dng cnh, mu xanh lc m khi
cn sng, khi cht hoc khi c nh bng
ho cht th chuyn mu xanh en, chng
to thnh khm sng trn cc khi san h
hoc bm trn nn y cng

110

2. Loi Dysidea cinerea, thuc h Dysideidae

M t: mc thnh dng
khi trn rn san h, mu
sc thany i rt nhiu
ph thuc vo mi trng
sng, t mu nu nht
cho ti mu xm hoc,
mm v dai, thng ln
c ct hoc sn bn trong
c th.

3. Loi Haliclona sp, thuc h Chalinidae

M t: thng c dng
cnh, cu trc thay i
tu loi c th mm hoc
hi cng, mu sc thay
i rt nhiu, t mu xm
nht, hng, tm hoc xanh
m, vi l m trn b
mt tng i ln
4. Loi Mycale plumosa, thuc h Mycalidae
M t: c dng cnh c
trng v thng ni vi
nhau, mu sc t m
cho ti mu tm nht,
thng b mt mu rt
nhanh sau khi ly ra khi
mi trng sng, cu trc
tng i mm

111

5. Loi Ircinia echinata, thuc h Irciniidae

M t: thng sng dng
khi c x thu, b mt
c bao ph bi cc mu
nh, thng c mu nu
en cho ti mu en, c
th do v n hi ging
nh cao su

6. Loi Amorphinopsis excavans, thuc h Halichondriidae

M t: y l loi c l
vo trong cc khi san h
hoc nn y v chim c
gn ht khng gian bn
trong, mu sc thay i
t vng cho ti vng m,
c l m trn b mt kh
ln. Khi kh thng
chuyn mu trng ng

7. Loi Clathria vulpina, thuc h Microcionidae

M t: c th dng cnh, c mu
vng cam, b mt x x c to
nn bi nhiu nm gai nh

112

8. Loi Petrosia nigricans, thuc h Petrosiidae

M t: thng c dng
khi, ging hnh ni la,
cu trc cng v d v, d
gy, b mt tng i
nhn, thng c mu tm
nht cho ti nu m

9. Loi Xestospongia testudinaria, thuc h Petrosiidae

M t: hnh dng, mu
sc v kt cu b mt
thay i rt nhiu tu
thuc vo mi trng
sng,

thng chng c

dng cc ln vi b mt
nhiu cc g v cc rnh,
cu trc cng v d v
vn.

10. Loi Niphates sp, thuc h Chalinidae

M t: c th dng cnh
phn nhnh, mu sc thay
i th mu trng ng
cho ti mu hng nht, b
mt tng i nhn, c
mt s l m nh trn b
mt.

113

11. Loi Stylissa flabelliformis, thuc h Axinellidae

M t: mc thnh cm
trn rn san h, c mu
vng cam, b mt x x
thng c nhiu mu nh
hoc c cc ng g
hoc np gp

12. Loi Ianthella sp, thuc h Ianthellidae

M t: C th c dng
qut, mu sc thay i t
mu vng nht, mu vng
cam m cho ti mu tm
sm, khi tip x vi
khng kh thng chuyn
mu tm sm

13. Loi Ircinia sp, thuc h Irciniidae

M t: thng c dng x
thu, c mu t nu m
ti mu en, i khi c
bao ph bi cc mu tht
nh, cu trc do v dai
ging cao su

I.5.3. Cc loi da gai

1. Hi sm Holothuria scabra, thuc h Holothuridae
c im nhn dng
L loi c c th ln, chiu di c th t n 30
cm. Bnh thng c th hnh qu chui, nhng c
th thay i rt nhanh nu mi trng c bin
di. C th mu en hoc en hi chuyn mu
nu, mu hng. Trn c th c hai dng gai hnh
gn trn v thoi bao ph
Phn b: Phn b rng dc theo di ven b
Vit Nam, tp trung ti Qung Ninh, Hi Phng,
ven bin min Trung

114

2. Sao bin cnh dy Protoreaster nodusus, thuc h Oreasteridae

c im nhn dng: Hnh sao 5 cnh, di cc
nh u nhau v tng i ngn. Trn mi cnh c t
- 6 hnh trn. Mu sc bin i t mu vng chanh
ng vng nht
Phn b: C Lao Chm, C T, trong cc rn
san h, cc bi ct ven b su 5 - 10 m
Sinh hc v sinh thi hc
Sao bin n to a bo nh: Sargassum,
Glacilaria, n cc loi ng vt y nh .a
mn cao t 20 - 30%0. Sng ven o, vng di
triu t 1 - 10 m nc, bm vo cc rn

I.6. Bc u xc nh cc khu vc c kh nng khai thc

ngun dc liu bin
Cc kt qu thu thp mu vt ca chng ti dc theo chiu di ca bin
Vit Nam, cho php bc u xc nh cc sinh cnh chnh m chng ta
c th thu thp cc loi sinh vt bin c gi tr lm dc liu
1. Cc tng, ng thuc qun o vi nh H Long - Ct B: Thng c
nhiu loi thuc nhm Hi min v sao bin, hi sm, phn b. khu vc ny
chng thng tp trung phn b thnh cc bi ln c gi tr khai thc. y l
khu vc c bit cn lu , trong tng lai, chng c th l nhng a im quan
trng cho vic nui trng v khai thc cc loi c cha dc liu bin. iu ny
chng t khu vc H Long -Ct B - Bi T Long l nhng ni c cc loi sinh
vt cha dc liu tp trung sinh sng .
2. Rn san h: l sinh cnh quan trng, y c th thu c rt nhiu
cc loi sinh vt c kh nng cha dc liu, c bit l nhm Da gai v san h
mm v Hi min. Cc rn san h thuc ng Nam Ct B, Hang Trai- u B,
C T, Hi Vn - Sn Ch, C Lao Chm, ven b Ninh Thun. c bit 3 khu
vc l Cn C, Cn o v Trng Sa l khu vc c mt cao ca cc loi san
h mm
3. Cc bi triu ct hoc ct pha v sinh vt, ct si: Phn b hu ht
cc cung lm ca cc o hoc ven b Vit Nam. y c th thu c hu ht
cc loi da gai, hi min c gi tr dc liu
Cc kt qu nghin cu ca chng ti cng xc nhn c 6 khu vc l
cc bi dc liu ln ca Vng bin Vit Nam
I.6.1. Bi dc liu ng nam o Ct B
Cc kt qu nghin cu xc nhn khu vc ng Nam o Ct B l ni
c a dng sinh hc cao ca nc ta ( Cng Thung v cc tc gi, 2002). T
y chng ta khng ch khai thc c nhng loi sinh vt c gi tr kinh t cao
m cn nhiu loi c gi tr dc liu cng phn b khu vc ny. Cc kt qu
nghin cu t nm 2003 n nay ca chng ti xc nh c cc im c
mc tp trung cao ca cc loi cha dc liu l Vn Bi, Vn H, Ba Tri
o, Hang C, Hang Ti. Cc bi c kh nng khai thc dc liu gm bi sao
bin, bi hi min cnh v km theo cc loi san h mm

115

- Bi Vn Bi - Vn H (Bi Sao bin): C din tch rng khong 5 ha

(50.000 m2), y l ni tp trung ca cc loi sao bin rnh nng v sao bin
rnh su (Astropecten polyacanthus, A. monacanthus) vi mt cao t 15 - 25
con/m2 (trung bnh 20 con/m2) bnh qun t n 0,3 kg/m2 (vo thng 3). Vi
khi lng ny, ring bi Vn Bi, Vn H c tr lng t 15 tn sao bin v
cho kh nng khai thc 7,5 tn/nm. Ngoi sao bin cn c th khai thc cc loi
Hi min cnh (Gellius varius), hi sm en (Holothuria martensii), san h mm
(Sarcophyton tortuosum), cu gai (Diadema setosum).
- Bi Tng Gu - Tng Hi: Bi cc nhm sao bin rnh su (Astropecten
polyacanthus) v hi min cnh (Gellius varius) cng vi nhiu nhm sinh vt
phn b. Din tch c khu vc khong 20 ha v c th khai thc c nhiu loi
sinh vt c gi tr dc liu (khng k hai loi nu trn) nh loi hi min khc
nh Ircinia echinata, Dycidea cinerea, D. fragilis, Petrosia nigricant, Mycale
plumosa, cu gai (Diadema setosum).
- Bi Ba Tri o (Bi hi min cnh): Bao gm ton b khu vc Ba Tri
o, hang Ti, hang Sng, hang C v tng Gi vi din tch khong 10 ha c
nhiu hi min cnh phn b (Gellius varius). Kt qu kho st s b xc
nh sinh khi t 0,08 - 0,1 kg/m2, tr lng t khong 8 - 10 tn. Ngoi loi
ch o hi min cnh, chng ti cng xc nh nhiu loi sinh vt c ngha
dc liu khc nh cu gai (Diadema setosum), hi sm en (Holothuria
martensii) v hng lot loi hi min khc nh Ircinia echinata, Dycidea
cinerea, D. fragilis, Petrosia nigricant, Mycale plumosa.v.v.
I.6.2. Bi dc liu khu vc vnh H Long
Cng nh Ct B, H Long l ni c nhiu cc tng, ng, cc rn san h
pht trin v cng l ni c nhiu loi sinh vt c kh nng cha dc liu pht
trin, to thnh cc bi quan trng. Trong cc nm 2003, 2004, 2006, 2007 cng
vi tp th cc nh khoa hc Italia, chng ti kho st, nh gi cc gi tr a
dng sinh hc ca khu vc ny. c bit, ln u tin chng ta c c kt qu
kho st tng i y v qun x hi min (Sponge) Vnh H Long. Trong
khu vc ny, chng ti thu thp c 27 loi hi min nm trong 13 h thuc
lp Desmospongia. Phn ln cc mu thu c u c tin hnh nh gi
hot tnh tm kim cc mu c hot tnh. Trong s cc loi hi min chim
u th l Mycale plumosa, Cliona celata, Cliona orientalis, Dysidea cinerea,
Gellius cf. varius, Aka mucosa, Ircinia echinata .v.v.
S pht trin mnh ca nhm hi min to ra kh nng thu thp mt
lng ln phc v cho nghin cu v chit xut cc cht t hi min. T nhng
kt qu thu nhn c, chng ti xc lp c 4 khu vc c kh nng khai thc
cc loi sinh vt ny l Vng Vua - Cng L, ng Nam o Hang Trai, Khu h
Ba Hm
- Khu vc Vng Vua - Cng L: L cc vng kh kn, nm ng nam
qun o u B; Din tch vo khong trn 100 ha. Cc loi sinh vt thu c
y gm hi min cnh (Gellius varius), hi min xp (Ircinia echinata), hi

116

min chum xanh (Biemna cf. megalosigma), hi min hnh ti (Xestospongea

testudinaria), hi min l (Aka mucosa), hi min bt (Dysidea cinerea) , hi
min dy (Mycale plumosa) v.v.. Ti y cng l ni c rn san h pht trin tt,
v vy ngoi san h cnh (Acropora nobilis), cn c th thu c san h mm
(Sarcophyton tortuosum), cu gai (Diadema setosum).

Hnh I.6.2. Cc bi dc liu bin ch yu khu vc H Long - Ct B

Ghi ch: 1- Bi Vn Bi - Vn H (Bi sao bin); 2- Bi Tng Gu - Tng Hi (Bi hai mnh v); 3- Bi
Ba Tri o (hi min cnh); 4- Bi Vng Vua - Cng L; 5- ng Nam o Hang Trai; 6- Khu H Ba
Hm

117

- Khu vc vc ng Nam o Hang Trai: C din tch khong gn 80 ha,

chy t pha Bc o u B n sn pha ng ca o Hang Trai. Cc loi
sinh vt thu c y gm hi min cnh (Gellius varius), hi min xp
(Ircinia echinata), hi min chum xanh (Biemna cf. megalosigma), hi min
xanh (Haliclona clathrata), hi min bt (Dysidea cinerea), hi min dy
(Mycale plumosa), cu gai (Diadema setosum).
- Khu h Ba hm: Nm pha bc o Hang Trai vi kh nhiu h nc
mn phn b. Ti y ngoi mu ca cc nhm Hi min nh hai khu vc trn
chng ti cng thu mt lot loi khc nh hi min bn (Gellioides fibulata),
hi min tm (Haliclona subarmigera), san h mm mt gai (Cladiella
pachyclados) v.v.
I.6.3. Bi dc liu khu vc vnh C T - Thanh Ln
C T - Thanh Ln l o xa b, nm pha ng Bc Vit Nam v c
tim nng bo tn cao vi cc c im l a dng v loi cao v c nhiu loi
qu him. Cc t kho st ca chng ti vo cc nm 2003, 2004, 2005, 2006
cng xc nh y l ni tp trung nhiu loi c gi tr dc liu cao. Trong s
cc loi sinh vt bin thu thp xc nh dc tnh th c ti 4 nhm c kh
nng tp trung thnh cc bi sinh vt c cha dc liu c gi tr, bao gm bi
trai ngc, bi sao bin, bi san h + cu gai + hi sm (hnh I.6.3).
- Bi sao bin: nm gn khu vc t Bc Vn n Hn Khi. Ti y xut
hin nhiu loi sao bin vi mu sc rc r. Cc loi thng gp gm sao bin
cnh dy to (Asterope carinifera), sao bin mnh dy (Asterina bacheri), sao bin
9 cnh (Ludia macculata), sao bin 5 cnh hng (Protoreaster nodusus). Ngoi
ra y cng c th thu c cc loi cu gai (Diadema setosum), hi sm en
(Holothuria vagabunda) v mt lot loi hi min (Gellius varius, Haliclona
clathrata, Dysidea cinerea, Thethya seychellensis .
- Bi san h, cu gai, hi sm : Xt trn kha cnh dc liu khu vc chy
t Bi Hng Vn n hn Khe Tru l quan trng nht. y, mi ngy ngi
dn a phng khai thc c t 20 - 50 kg hi sm en (Holothuria vagabunda)
mang bn, bnh qun khong 30 kg /ngy, mt thng thu 900 - 1000 kg hi sm.
Hi sm en thng sng trn cc rn san h v theo nc triu tp vo cc bi
ct kim n. Trn mi mt ct di 50 m, rng 3 m chng ti thu c t 20 - 30
con hi sm nh vy c khong 5 - 10 m2 tm thy 1 con hi sm sng. Ngoi hi
sm y cn l ni tp trung ca cu gai (Diadema setosum). Cu gai phn b vi
mt cao trn cc rn san h ti Hng Vn, thm ch c ni t n 5 - 10
con/m2. mt ny hon ton cho php chng ta khai thc mt lng mu ln
hng tn tin hnh chit xut cc cht cn thit. Cc kt qu nghin cu cn
xc nh rn san h Hng Vn, Khe Tru l ni c san h cnh pht trin tt nht
nh vnh Bc B. T san h cnh, c th chit xut cc cht c gi tr dc liu
tt.
Ngoi 3 loi chnh k trn ti y cn c th thu c cc loi c gi tr
nh bn mai Qut (Atrina vexillum), da chut bin (Stichopus sp), Hi min nh
Gellius varius, Haliclona submarginea, Haliclona clathrata, Dysidea cinerea.

118

Hnh I.6.3. Cc bi dc liu bin ch yu khu vc C T - Thanh Ln

Ghi ch: I- Bi sao bin; II- Bi Trai ngc; III- Bi san h, cu gai, hi sm

I.6.4. Bi dc liu Cn C
Tuy l o nh nhng hu ht xung quanh o u c rn san h phn
b. Trn cc rn san h cng c nhiu qun th san h mm v cu gai sinh sng.
Trong t kho st thng 11/2006, chng ti pht hin s tp trung cao ca
san h mm pha ty - bc o, trn din tch khong 5 ha. y c th khai
thc cc loi san h mm Cladiella, Sarcophyton trocheliophorum, Lobophyton

119

crassum, Lobophytum sp, Lobophytum compactum, Klyxum sp vi tr lng 5 10 tn ( theo phng vn dn a phng)

Hnh 1.6.4. Bi dc liu Cn C

I.6.5. Bi dc liu Cn o
y l khu bo tn bin quan trng ca Vit Nam. Ti y cha ti 1351
loi ng, thc vt bin (.C. Thung, 2007) , trong s ny c trn 70 loi Da gai
v khong 5 loi san h mm. Khu vc ca mc tp trung cao, to thnh cc
bi dc liu l khu vc Hn By cnh, hn Trc v vnh Cn Sn . nhng chng
ti cha c dn liu v tr lng khu vc ny
120

Bn cc bi dc liu ch yu khu vc Cn o
670000

675000

680000

685000

690000

695000

2,500

970000

970000

hn Trng

5,000
Meters

965000

965000

hn Tre Nh
0

hn Tre Ln
o Cn Sn

hn Cau

960000

960000

hn By Cnh

0
0

0 hn Bng Lan

hn Tho
0

hn Vung 0

955000

955000

Cc bi dc liu (hn Trc, vnh Cn Sn, hn By Cnh)

670000

675000

680000

685000

690000

695000

Hnh 1.6.5. Bi dc liu Cn o

I.6.6. Bi dc liu Trng Sa
Hu ht cc o thuc qun o Trng Sa u c cc rn san h phn b.
c bit hai o Thuyn Chi v Nam Yt c din tch t 20 n hng trm km2
rn san h. y l ni tp trung cao ca cc loi san h mm v sao bin, cu gai.
Ti y c th khai thc mt tr lng ln cc loi sinh vt cha dc liu, nhng
chng ti cha c c hi kho st ni dung ny.

121

Hnh 1.6.6. Bi dc liu Nam Yt

I.7. xy dng C s d liu sinh vt bin
Cc d liu v sinh vt bin c lu d di 4 dng sau:
- Cc mu thu c c xc nh c h s mu km theo, c lu tr
trong phng th nghim so snh
- H s ca 101 mu thu (t thu mu, thi gian thu mu, a im v
tn loi thu c)
- nh ca tt c 101 mu thu c lu tr bng 101 file dng jpg, c
th x l nhiu phn mm khc nhau
- M hnh c s d liu qun l cc loi sinh vt bin c cha dc liu
I.7.1. Mc ch
Thit k, xy dng c s d liu sinh vt bin cha dc liu l h thng
chng trnh h tr cng tc qun l, tra cu, tm kim thng tin v mt s i
122

tng c cha dc liu sng vng bin ven b nh San h mm, da gai, hi
min... Cc loi sinh vt ny c thu mu trong cc nm 2006 - 2008 mt
s vng c trng ven bin nh Ct B, Cn C, Trng Sa, Cn o v Vnh H
Long. C s d liu c thit k n gin, tin li v d s dng. Gip cc nh
khoa hc, nh qun l u c th d dng Cp nht, hiu chnh, lit k, b sung
thng tin v cc loi sinh vt cha dc liu vo c s d liu, tm kim v tra
cu thng tin, in n v truy xut thng tin kt qu ra my in.
C s d liu sinh vt bin
c kh nng cha dc liu

Mu lu tr

H s mu

B nh v m
hnh CSDL

Hnh I.7.1. S cu trc lu tr d liu cc loi SVB c cha dc liu

I.7.2. Phng php thit k v xy dng m hnh CSDL sinh vt bin cha
dc liu
C s d liu c thit k v xy dng da trn m hnh lin kt gia phn
mm Microsoft Access v Visual Basic cng vi s h tr ca cc phn mm
chuyn dng khc nh Photoshop (x l nh), Microsoft Office (xy dng cc
file dng text) ... Cc bc tin hnh xy dng c s d liu gm hai cng on
(hnh I.7.2.a)
Thu thp thng tin CSDL cc loi sinh vt bin cha dc liu

Xy dng CSDL sinh vt bin cha dc liu

Hnh I.7.2.a. S m t cc bc thc hin

Thu thp mu vt v thng tin
thu thp 101 mu sinh vt bin phn b dc di ven b Vit Nam. Cc
mu c chp nh v tin hnh phn loi mu. Mt s loi ch yu c m t
chi tit cc c im nhn dng, phn b v gi tr ca mi loi
Xy dng c s d liu
Phn tch v thit k h thng cho Metadata C s d liu a dng sinh hc
cc loi sinh vt bin cha dc liu:

123

Bao gm cc m hnh qun l cc tp thng tin v c s d liu a dng loi sinh

vt bin cha dc liu cho mi vng v cc thng tin m t ca tng loi (hnh
I.7.2.b)
CSDL a dng sinh hc cho cc vng

Thng tin m t tng loi

Hnh I.7.2.b. M hnh tng qut c s d liu sinh vt bin cha dc liu
Nh vy mi lin kt gia c s d liu v ngi s dng c thc hin nh
cc mdun x l thng tin. Cc lnh truy xut hoc thc hin mt chc nng no
c truyn n m hnh c s d liu v cc thng tin cn thit c trch
rt theo yu cu v a n ngi s dng thng qua cc modun x l thng tin
(hnh I.7.2.c).
Giao din ngi s dng

Cc Modun
x l thng tin

C s d liu
cc loi sinh vt bin
cha dc liu

Hnh I.7.2.c. H thng qun l c s d liu sinh vt bin c cha dc liu

c th chun ho cc thng tin v c s d liu ca tng loi, chng ti
xy dng cc trng thng tin cho tng bn record. Mi bn record gm 9 trng
thng tin m t cc chc nng khc nhau (bng I.7.2)
Bng I.7.2. M t cc trng d liu
STT
Tn trng
1
loai-id
2
loai-name-viet

M t
M s loi
Tn loi ting Vit Nam

124

3
4
5
6
7
8
9

loai-name-nc ngoi
loai-synonim
loai-dacdiemnhandang
loai-phanbo
loai-sinhhocsinhthai
loai-picture
loai-giatri

Tn khoa hc
Synonim ca loi
M t c im nhn dng
a im Phn b
c im sinh thi ca loi
Hnh ca loi
Gi tr dc liu

M t kh nng truy xut thng tin t c s d liu a dng sinh vt bin

trn hnh I.7.2.d.
Ngi s dng

Yu cu

Tr li

Giao din ngi s dng

Kim tra yu cu ca
ngi s dng

Khi to chc nng

X l

Hin th thng tin

Cp nht
Tm kim
B sung
Hiu chnh
Hu b
In n thng tin
...

C s d liu
a dng sinh hc

Hnh I.7.2.d. S tng qut chu trnh x l thng tin

Chun ho v qun l thng tin v CSDL a dng sinh hc ca tng loi

125

- Trng loai-id: M s ng k ca loi trong c s d liu, c ng

theo s th t t 0 - ht
- Trng loai-name-viet : Tn ting Vit ca loi nghin cu
- Trng loai-name-nuoc ngoai: Trng ghi tn khoa hc ca loi
- Trng loai- synonim: Trng ghi tn ng ngha ca loi
- Trng loai-dacdiemnhandang: M t cc c im nhn dng ca loi. S
lng k t m t khng hn ch
- Trng loai-phanbo: Cho php cp nht cc m t phn b ca tng loi,
s lng k t khng gii hn
C s d liu c th qun l n tng loi, cho php sa cha tng bn
record theo tng a danh v tng nhm sinh vt khc nhau (hnh I.7.2.e)
CSDL a dng sinh hc

Qun l cc thng tin v

cc i tng

Cp nht - B sung - Sa cha

Tm kim - Hu b ca cc vng

Cn C

H Long
Ct B
Trng Sa
Cn o

Hnh I.7.2.e. Cu trc bng CSDL theo a im

- Trng loai-sinhhocsinhthai: Cp nht cc c im v sinh thi ca loi,
s lng k t khng gii hn
- Trng loai-picture: Cho php cp nht nh t th mc picture vo c s
d liu
- Trng loai-giatri: Cp nht cc gi tr ca loi vo c s d liu, s lng
k t khng hn ch
I.7.3. Cc chc nng c s d liu c th thc hin
H thng c s d liu xy dng da trn phn mm Visual Basic v
Microsoft Acess cho php gim thiu cng sc, thi gian ca ngi s dng,
cng nh gip cho cng tc qun l, thng k v khai thc thng tin c thc
hin d dng v hiu qu hn. H thng c th m rng, kt ni vi cc CSDL
khc v cho php ngi s dng b sung cc tnh nng mi nu cn.

126

 Hin ti chng trnh c a vo khai thc v s dng ti Phng Sinh

thi Ti nguyn Sinh vt bin, Vin Ti nguyn v Mi trng Bin. H thng
m bo tnh chnh xc v thun tin.
cp nht c thng tin v mt s i tng sinh vt bin thuc cc
nhm san h mm, da gai v hi min trong mt s vng c trng ven bin (C
T, H Long, Ct B, Qung Tr, Cn o, Trng Sa).
Cc chc nng chnh H thng h tr ngi s dng qun l thng tin a
dng sinh hc:
- Cp nht thng tin
- B sung thng tin
- Hiu chnh thng tin
- Hu b thng tin
- Tra cu thng tin
- In n thng tin
I.8. Nhn xt v xut phng hng nghin cu tip theo
I.8.1. Nhn xt
- nh hng nghin cu tp trung vo 3 nhm sinh vt da gai, san h
mm v hi min nhm tm kim c cc cht hot tnh c dc l c bit l
hon ton chnh xc
- Cc loi sinh vt m chng ti thu c c vng phn b rng hu ht
cc o ven b cng nh xa b Vit Nam
- Nhiu loi sinh vt thu c c sinh khi cao, c kh nng to ra ngun
dc liu t nhin sn xut ra cc loi thuc qu
I.8.2. xut
- Hon thin nghin cu phn b ca cc loi c gi tr dc liu cao
- M rng cc i trng nghin cu trong nhm sao bin, hi sm mt,
hi sm v l nhng loi c dn coi l nhng v thuc i b bin
- Nn xem xt kh nng to ra cc sn phm c gi tr dinh dng cao t
cc loi sinh vt bin

127

Chng II. x l mu, to dch chit v sng

lc hot tnh sinh hc
Tt c cc mu sinh vt bin thu thp c trong ti u c x l s
b, bo qun v to dch chit th theo quy trnh thng nht nh trnh by
trong phn phng php nghin cu. Cc dch chit c tin hnh sng lc hot
tnh sinh hc theo nh hng: khng vi sinh vt kim nh, chng xy ha v
gy c t bo. Di y l kt qu c th:
II.1. Kt qu to dch chit
Sau khi chit xong ton b mu, cc s liu ca qu trnh chit mu (K
hiu mu, lng mu chit, lng cn dch thu c) c tp hp li thnh
bng v tnh hiu sut thu hi cn dch.
Bng II.1. Kt qu to dch chit cc mu sinh vt bin

TT

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19

K
hiu
mu

TnVit Nam

Tn khoa hc

Gellius varius

KC-002
KC-003
KC-004
KC-005
KC-006
KC-007
KC-008

Hi min cnh
xanh
Hi min
Hi min
Hi min
Hi min
Hi min
San h mm
San h mm

KC-009
KC-010
KC-011

San h mm
San h mm
San h mm

KC-012

San h mm

KC-013
KC-014
KC-015
KC-016
KC-017

Hi min
San h mm
San h mm
Hi min
Hi min hnh
nn
Hi min hnh
chung
Hi min hnh

KC-001

KC-018
KC-019

Dysidea cinerea
Haliclona sp
Mycale plumosa
Ircinia echinata
Amorphinopsis excavans
Claudiella sp
Sarcophyton
trocheliophorum
Lobophyton crassum
Lobophytum sp
Lobophytum
compactum
Lobophytum
compactum
Amorphynopsis sp
Sinularia abrupta
Klyxum sp
Petrosia nigricans
Ianthella sp
Xestospongia
testudinaria
Xestospongia

128

Lng
mu

chit
(g)

Lng
cn
dch
chit
(g)

%
Thu
hi
cn
dch
chit

500

2,75

0,55

500
500
500
500
1000
1000

2,45
2,80
3,70
4,25
4,00
9,60

0,49
0,56
0,74
0,85
0,40
0,96

1000

13,40

1,34

1000
1000

11,80
15,50

1,18
1,55

1000

16,50

1,65

1000

10,80

1,08

1000
1000
500
500

8,80
12,20
7,15
3,30

0,88
1,22
1,43
0,66

500

2,85

0,57

1000

8,30

0,83

500

4,25

0,85

23

KC-023

chung
Hi min
mng mng
San h mm
Hi min cy

San h mm

24

KC-024

San h mm

Lobophytum compactum

KC-025

San h mm
hnh l gan

Sarcophyton mililatensis

26

KC-026

San h mm

Lobophytum compactum

1000

13,90

1,39

27

KC-027
KC-028

Hi min
Hi min xp

500

3,35

0,67

1000

8,20

0,82

KC-029
KC-030

Hi sm en
Hi min hnh
qu u
Da gai ui
rn
Hi min xp
l
San h mm
hnh chn vt
Hi min cnh

Hi min
phin
San h mm
hnh chm
chm
Hi min cnh
Da gai hnh
trng
San h mm
San h mm
San h mm
Hi min
hnh chung
C gim gai di
Hi min
Hi min xanh
Hi min hnh
ti
Hi min
Sao bin
Hi min

Petrosia nigricans
Xestospongia
testudinaria
Holothuria atra
Niphates sp

500

7,60

1,52

500

3,95

0,79

500

3,25

0,65

500

2,05

0,41

1000

14,50

1,45

500

2,20

0,44

500

1,85

0,37

500

4,90

0,98

500

3,00

0,60

500

3,85

0,77

500
1000
500

8,90
16,60
5,35

1,78
1,66
1,07

500

2,60

0,52

500
500
500

5,20
2,80
2,20

1,04
0,56
0,44

1000

7,70

0,77

500
1000
500

2,25
10,60
1,15

0,45
1,06
0,23

20
21
22

25

28
29
30
31
32
33
34
35

KC-020
KC-021
KC-022

KC-031
KC-032
KC-033
KC-034
KC-035
KC-036

36
37
38
39
40
41
42
43
44
45
46
47
48
49

KC-037
KC-038
KC-039
KC-040
KC-041
KC-042
KC-043
KC-044
KC-045
KC-046
KC-047
KC-048
KC-049

testudinaria
Ircinia sp

500

3,75

0,75

Lobophytum compactum
Clathria vulpina

1000

12,10

1,21

500

3,45

0,69

Sarcophyton elegans

500

7,75

1,55

500

4,85

0,97

1000

10,90

1,09

Macrophiothrix sp
Spongia sp
Sinularia sp
Clathria vulpina
Stylissa flabelliformis
Dendronephthya
klunzeringi
Gellius var fibrosa
Echinocardium
cordatum
Sarcophyton sp
Lobophytum sp
Lobophytum sp
Haliclona subarmigera
Diadema setosum
Ircinia echinata
Biemna cf. nudelringna
Xestospongea
testudinaria
Gelliodes fibulata
Anthenea pentagonala
Aka mucosa

129

50
51
52

KC-050
KC-051
KC-052
KC-053

53
KC-054

54
KC-055

55
56
57
58
59
60
61
62
63
64
65
66

KC-056
KC-057
KC-058
KC-059
KC-060
KC-061
KC-062
KC-063
KC-064
KC-065
KC-066
KC-067

67
KC-068

68
KC-069

69
KC-070

70
71
72
73
74
75
76
77
78
79
80

KC-071
KC-72
KC-73
KC-74
KC-75
KC-76
KC-77
KC-78
KC-79
KC-80

Hi min bt
Hi min trng
Hi min tm
Sao bin nm
cnh mu
hng
Sao bin 5
cnh mu tm

Sao bin 5
cnh mu tm
ru
Sao bin rnh
nng
Da chut bin
Hi min xp
Hi min xp
Hi min
Hi sm
Hi min hnh
chp
Hi min hnh
chp
Hi min
San h mm
San h mm
Sao bin 5
cnh mu tm
ru
Da chut bin
mu hng c
gai
Da chut bin
mu hng
khng gai
Sao bin cnh
ngn mnh dy
mu hng m
Sao bin
San h mm
San h mm
San h mm
San h mm
San h mm
San h mm
San h mm
San h mm
San h mm

Dysidea cinerea
Mycale plumosa
Haliclona submarginera
Protoreaster nodusus

500
500
500

1,95
2,80
2,35

0,39
0,56
0,47

500

5,80

1,16

1000

15,50

1,55

1000

11,30

1,13

1000

9,30

0,93

500
500
500
500
1000

8,55
1,65
2,10
3,05
12,50

1,71
0,33
0,42
0,61
1,25

500

2,80

0,56

500

2,60

0,52

500
1000
1000

1,95
11,90
10,20

0,39
1,19
1,02

1000

11,60

1,16

500

5,90

1,18

500

5,85

1,17

1000

10,80

1,08

100
500
1000
1000
500
500
500
500
500
1000

1,32
5,85
12,20
10,80
5,50
7,15
5,00
4,80
6,20
10,20

1,32
1,17
1,22
1,08
1,10
1,43
1,00
0,96
1,24
1,02

Asterope carinifera
Asterope carinifera
Astropecten
polyacanthus
Holothuria spinifera
Dysidea cinerea
Biemna megalosigma
Ircinia echinata
Holothuria scabra
Petrosia nigricans
Petrosia nigricans
Spongea sp
Cladiella sp
Sarcophyton sp
Asterope sp
Stichopus sp
Thyone bicornis
Asterina batheri
Archaster typicus
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp

130

81
82
83
84
85
86
87
88
89

KC-81
KC-82
KC-83
KC-84
KC-85
KC-86
KC-87
KC-88
KC-89
KC-90

90
KC-91

91
KC-92

92
93

KC-93
KC-94

94
95
96

KC-95
KC-96
KC-97

97
KC-98

98
KC-99

99
100 KC-100
101

KC-101

Hi sm en
Sao bin
Sao bin
Cu gai
Cu gai
C ghim
Sao bin
San h mm
Hi sm en
Hi min cnh
xanh
Sao bin
Sao bin
Hi min
Hi min hnh
ti
Hi min cnh
Hi min xanh
Hi min
trng
Sao bin
Sao bin
San h mm
Sao bin rnh
nng

Holothuria vagabunda
Astropecten polycanthus
Astropecten polycanthus
Echinothrix calamaris
Echinothrix calamaris
Diadema setosum
Astropecten polycanthus
Sarcaphyton sp
Holothuria vagabunda
Gellius varius
Astropecten
polyacanthus
Astropecten
monacanthus
Biemna cf. nudelringna
Xestospongea
testudinaria
Gelliodes fibulata
Haliclona clathrata
Mycale plumosa
Astropecten
polyacanthus
Astropecten
monacanthus
Lobophytum sp
Astropecten
polyacanthus

1000
1000
500
500
500
1000
500
1000
1000

10,50
10,20
5,55
4,70
4,50
8,50
5,85
12,60
11,80

1,05
1,02
1,11
0,94
0,90
0,85
1,17
1,26
1,18

500

2,75

0,55

1000

11,00

1,10

500

6,95

1,39

500

2,40

0,48

500

3,00

0,60

500
500

1,60
4,45

0,32
0,89

500

5,05

1,01

1000

11,30

1,13

1000

10,70

1,07

500

6,20

1,24

500

5,85

1,17

II.2. kt qu nh gi hot tnh khng vi sinh vt kim nh

(antimicrobial activity)
101 Mu sinh vt bin thuc nhm hi min, da gai v san h mm c
th nghim v nh gi hot tnh khng vi sinh vt kim nh trn cc dng vi
sinh vt gy bnh tiu biu nht thuc cc nhm vi khun Gr(-); vi khun Gr(+);
nm mc v nm men. nh gi s b hot tnh, chng ti pha mu nng
theo tiu chun sng lc s b l 4 mg/ml v tm cc mu th hin hot tnh
khng vi sinh vt kim nh nng . Cc mu c biu hin dng tnh
bc s b ny c pha long theo cc thang nng thp dn, t 5-10 thang
nng tnh gi tr nng ti thiu (MIC) m vi sinh vt b c ch pht
trin gn nh hon ton. Ngoi ra, cc mu c biu hin dng tnh cn c tin
hnh chit phn b gia dung mi hu c v nc theo phn cc ca dung mi

131

tng dn t hexan, sang clorofooc, butanol. Cc dch chit ny c tin hnh xc

nh gi tr MIC nh hng cho cc nghin cu tip theo. Kt qu gi tr nng
c ch ti thiu MIC c tng kt trn Bng 1.
Bng 1: Tng hp kt qu gi tr MIC ca cc mu sinh vt bin.
K hiu Dch
mu
chit

KC-001

KC-002

KC-003

KC-004

KC-005

KC-006

KC-007

KC-008

KC-009
KC-010
KC-011

KC-012

KC-013

M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
M
H
C
B
N
M
H
C
B
N
M
M
M
M
H
C
B
N
M

Vi khunGr(-)

Nng c ch ti thiu(MIC: g/ml)

Vi khunGr(+)
Nm mc

Nm men

E.coli

P.aeru

B.subti

S.aureus

Asp.niger

F.oxy

S.cere

C.albic

150
(-)
100
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
(-)
150
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
(-)
50
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

200
(-)
150
150
(-)
150
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
50
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
50
50
100
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
100
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
(-)
150
(-)
100
50
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150

(-)
(-)
(-)
(-)
(-)
100
(-)
100
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
50
(-)
(-)
(-)
200
(-)
100
(-)
(-)
100
50
50
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
200
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
150
100
(-)
(-)
(-)
(-)
(-)
(-)
200
50
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

150
(-)
(-)
100
(-)
200
(-)
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
50
50
50
(-)
(-)
(-)
100
(-)
(-)
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
(-)
50
50
(-)
200
(-)
(-)
150
150
100
50
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
50
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

132

KC-014

KC-015
KC-016

KC-017

KC-018

KC-019

KC-020

KC-021
KC-022

KC-023

KC-024

KC-025

KC-026
KC-027

KC-028

KC-029

H
C
B
N
M
H
C
B
N
M
M
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
M
M
H
C
B
N
M
M
H
C
B
N
M
M
M
H
C
B
N
M
H
C
B

(-)
(-)
(-)
(-)
200
100
(-)
200
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
50
(-)
(-)
100
50
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
100
100
50
50
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
(-)
(-)
(-)

(-)
100
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
50
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
150
150
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
200
200
50
(-)
(-)
(-)
100
100
50
(-)
(-)
(-)
(-)
(-)
(-)

133

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
(-)
100

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
100
50
(-)
(-)
150
100
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
50
(-)
(-)
(-)
150
50
100
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
200
150
100
(-)
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
50
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
(-)
50

KC-030
KC-031
KC-032
KC-033
KC-034
KC-035
KC-036

KC-037

KC-038

KC-039

KC-040
KC-041

KC-042

KC-043

KC-044

KC-045

KC-046

KC-047
KC-048
KC-049

KC-050

KC-051

KC-052

N
M
M
M
M
M
M
M
M
H
C
B
N
M
M
H
C
B
N
M
M
M
H
C
B
N
M
M
H
C
B
N
M
M
H
C
B
N
M
M
M
M
H
C
B
N
M
H
C
B
N
M
H
C
B

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
100
50
(-)
(-)
(-)
150
100
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
100
200
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
100
150
(-)
(-)
200
(-)
150
50
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
150
50
(-)
(-)
(-)
150
200
100
(-)
(-)
(-)
(-)
(-)
100
100
(-)
50
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
50
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
100
100
(-)
50
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
200
50
(-)
(-)
(-)
(-)
(-)
100
100
200
100
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

134

150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
(-)
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
150
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
(-)
100
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
150
100
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
(-)
(-)
100
150
200
100
(-)
(-)

KC-053
KC-054
KC-055

KC-056

KC-057

KC-058

KC-059

KC-060

KC-061

KC-062
KC-063
KC-064

KC-065

KC-066

KC-067

KC-068

KC-069
KC-070

N
M
M
M
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
M
M
M
H
C
B
N
M
H
C
B
N
M
M
H
C
B
N
M
M

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
150
100
200
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
(-)
150
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
100
150
200
(-)
200
100
150
(-)
(-)
150
50
50
200
(-)
150
100
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
100
(-)
150
(-)
200
(-)
150
(-)
200
(-)
150

(-)
(-)
(-)
(-)
200
(-)
(-)
100
200
(-)
(-)
(-)
(-)
(-)
150
100
100
150
(-)
100
50
50
200
(-)
100
50
50
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
50
(-)
200
(-)
(-)
(-)
(-)
(-)
150
50
50
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
(-)
100
(-)
100
50
100
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100

135

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
50
(-)
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
100
100
50
(-)
150
100
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
50
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
150
200
200
(-)
(-)
(-)
(-)
(-)
50
50
50
100
(-)
200
(-)
150
(-)
200
(-)
(-)
(-)
150
(-)
(-)
100
100
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
150
(-)
200
(-)
(-)

(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
50
50
150
(-)
(-)
100
100
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
(-)
200
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100

KC-071

KC-072
KC-073
KC-074

KC-075

KC-076

KC-077

KC-078
KC-079

KC-080

KC-081

KC-082
KC-083
KC-084
KC-085
KC-086

KC-087

KC-088

KC-089

H
C
B
N
M
H
C
B
N
M
M
M
M
H
C
B
N
M
H
C
B
N
M
H
C
B
N
M
M
M
H
C
B
N
M
H
C
B
N
M
M
M
M
M
M
H
C
B
N
M
H
C
B
N
M

(-)
(-)
(-)
(-)
200
(-)
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
50
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
100
100
50
(-)
(-)
(-)

(-)
100
100
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
150
200
(-)
(-)
100
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
150
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
(-)
100
(-)
(-)
(-)
(-)
(-)
(-)
200

(-)
(-)
(-)
(-)
200
(-)
100
50
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
(-)
(-)
100
100
(-)
(-)
(-)
(-)
(-)
(-)

(-)
(-)
100
150
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
200
200
(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
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50

(-)
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H
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C
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150
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B
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N
M
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H
KC-090
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C
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B
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N
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KC-091
M
M
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H
KC-092
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200
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C
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150
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B
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N
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KC-093
M
M
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H
KC-094
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C
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KC-095
M
M
100
15O
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H
KC-096
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C
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M
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M
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KC-101
M
Ch thch: M - Cn dch tng metanol, H - Cn
butanol, N - Cn chit nc.

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chit hexan, C - Cn

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chit clorofooc, B -

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100
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Cn chit

Kt qu sng lc trn bng 1 cho thy t l cc mu c hot tnh l kh

cao: c 51 mu trn tng s 101 mu khng t nht mt chng vi sinh vt kim
nh (chim t l 50,49%). Trong , s mu c hot tnh khng 1 vi sinh vt
kim nh l 8 mu (chim 7,92 %), khng 2 vi sinh kim nh l 10 mu, khng
3 vi sinh kim nh l 19 mu, khng 4 vi sinh kim nh l 10 mu, khng 5 vi
sinh kim nh l 2 mu, khng 6 vi sinh kim nh 1 mu v khng 7 vi sinh
kim nh l 1 mu. Kt qu c th hin qua bng 2.

137

60
50

T l %

40
30
20
10

g
h
n

M
u

V
7
K

h
n

h
n

V
SV
d
n
g
tn
h

SV

SV
V
6

h
n
K

h
n
K

h
n

2
g

SV
V

SV
V

SV
V

SV
V
1

h
n

g
K

h
n
K

SV

Biu 1: Biu thng k hot tnh khng s lng chng vi sinh vt kim nh
ca 101 mu sinh vt bin
Bng 2: Thng k hot tnh khng VSV kim nh ca 101 mu sinh vt bin
Tng s mu

Khng 1 Khng 2 Khng 3 Khng 4 Khng 5 Khng 6 Khng 7 Khng 8 S mu c

VSVK VSVK VSVK VSVK VSVK VSVK VSVK VSVK

hot tnh

101

10

19

10

51

T l %

7,92

9,90

18,81

9,90

1,98

0.99

0,99

50,49

nh gi mt cch r rng hn kh nng khng cc chng VSVK

khc nhau, chng ti tin hnh thng k, tnh ton s lng v t l % ca cc
mu sinh vt bin khng tng chng VSVK c th. Kt qu c a ra trong
bng 3.
Bng 3: Kh nng khng tng chng VSVK ca 101 mu sinh vt bin
Vi khun Gr (-)
E.coli

S mu c
hot tnh
T l % trn
101 mu th

Vi khun Gr (-)

P.aeruginosa B.subtillis

Nm mc

Nm men

S.aureus

Asp.niger

F.oxysporum

C.albicans

S.cerevisiae

15

31

13

21

10

15

17

18

14,85

30,69

12,87

20,79

9,90

14,85

16,83

17,82

138

So snh kh nng khng cc vi sinh vt kim nh ca cc mu c hot tnh

khng vi sinh kim nh c th hin trn biu 2.
35.00

30.69

30.00

T l %

25.00

20.79

20.00
14.85

15.00

14.85

12.87

16.83 17.82

9.90

10.00
5.00

E.
co
P.
li
ae
ru
gi
no
sa
B.
su
bt
i ll
is
S.
au
re
us
As
p.
ni
ge
F.
r
ox
ys
po
ru
m
C.
al
bi
ca
ns
S.
ce
re
vi
sia
e

0.00

Biu 2: Biu thng k kh nng khng cc vi sinh vt kim nh

ca 101 mu sinh vt bin
Kt qu trn cho thy, t l mu sinh vt bin khng cc chng P.
aeruginosa v S. aureus. c bit l vi chng P. aeruginosa, t l mu biu hin
hot tnh ln ti 30,69 %. y l trc khun m xanh thuc dng vi khun Gram
(-), hiu kh, c th gy bnh c hi cho ngi, c bit l nhng ngi c vn
v h thng min dch. N l loi in hnh nht ca chi Pseudomonas v cng l
loi vi khun c xc nh l nguyn nhn ch yu (khong 10%) gy ln cc ca
bnh nhim trng c ghi nhn trong cc bnh vin (hu ht cc ca nhim trng
khi b bng l do vi khun ny). Chng nhim vo nhng ngi b suy gim min
dch hay b bng, b thng v nhng ngi phi s dng ng thng hay my h
hp nhn to. Nhim khun Pseudomonas c th dn ti nhim trng ng tit
niu, nhim trng mu, vim phi, vim hng v nhiu bnh khc [159].
Pseudononas pht trin thnh khun lc trn phi ca bnh nhn b x nang
(cystic fibrosis - CF), v lm tng tnh trng mn tnh ha ca bnh v phi v
tng t l t vong khi b x nang. Vi khun Pseudomonas thng sinh sng trong
t, vng ly li v cc vng duyn hi. N cng c th sng st di cc iu
kin m t c loi vi khun no chu ng ni, n to ra mt lp mng mng c
th chng li s thc bo, v hin nay, chng khng hu ht cc loi khng sinh
ang c s dng. Pseudomonas c th sinh si c nhng iu kin mi
trng rt khc nhau nh: s phng, bn ra chn, thit b gy m v hi tnh, cc
loi nhin liu, my lm m v thm ch c trong nc ct c lu tr. N cng
c pht hin trong my chy thn. Mu xanh c trng ca m l do sc t
xanh (pyocyanin) v sc t lc (fluorescein) c sn sinh bi vi khun
Pseudomonas.
139

Ngoi ra, t l mu c biu hin hot tnh khng t cu khun Staphylococcus

aureus cng t t l 20,79 %. T cu khun c t l gy bnh rt cao, c kh
nng gy nhiu bnh nng cng nh khng khng sinh rt mnh. Cc nh vi
khun hc lng danh nh Robert Koch (1878) v Louis Pasteur (1880) u rt
quan tm nghin cu t cu khun ngay t thi k u ca lch s ngnh vi sinh
vt hc. Trn phng din gy bnh, t cu khun c chia thnh hai nhm
chnh: t cu c men coagulase (gm S. aureus v S. intermedius) v t cu
khng c men coagulase [160 - 161]. T cu c men coagulase gyln nhiu
bnh l khc nhau nh:
Cc bnh nhim trng khu tr ti da v nim mc: nhng bnh nhim trng
da v cc phn ph thuc (ch yu l cc chn lng v tuyn m hi) to thnh
bnh cnh p xe kinh in ca t cu. Cc t cu khun c kh nng to fibrin
do to c mt vch fibrin bao bc p xe. Cc nhim trng ny c th
ch nh nh u inh ghim (bnh vim nang lng) hoc kch thc nh qu to
trong p xe c. Cc vng da c lng rm bao ph, t cu thng l nguyn
nhn gy nn cc mn u anh. Tn thng ti ch c th nh nhng nhng
n cng l mt mi nguy c pht tn vi khun n nhng c quan xa hn. M
ca cc p xe do t cu vng thng c mu vng, c v khng hi.
Nhim trng cc c quan su: nhim trng cc c quan bn trong c th c th
do ng ni sinh: t mt nhim vim nhim ngoi vi, vi khun theo ng
mu v bch huyt n cc c quan khc. Nhim trng cng c th do nguyn
nhn ngoi sinh: vi khun i t mi trng vo c th thong qua vt rch da
sau chn thng hoc trong qu trnh phu thut. Cc bnh l in hnh trong
nhm ny c th k: vim xng ty xng, nhim trng huyt, vim tc tnh
mch xoang hang, vim ni tm mc, vim mng no m...
Vim da bng nc (bong da s sinh, hi chng bong da do t cu) thng gp
tr s sinh v tr nh. Tc nhn gy bnh l cc chng t cu sn xut c t
Exofoliatine. Ngay sau triu chng st v da, bnh c c trng bng du
hiu bong lp biu b ng thi hnh thnh bng nc trn din rng. Bnh
thng din tin lnh tnh. Mc d trn da hnh thnh nhiu bng nc ln
nhng t khi a n tnh trng ri lon nc v in gii. Lp da non mi
c hnh thnh kh nhanh.
Hi chng sc nhim c: thng gp ph n tr trong k kinh nguyt c
dng bng thm ht mnh. Chnh do kh nng thm ht tt ny m bng c
thi gian s dng lu hn. Mt s loi bng thm ny c kh nng gn vi cc
ion magnesium do lm gim lng ion ny trong m o. Khong 20% ph
n c mang t cu ng m o. Do lng ion magnesium gim xung, vi
khun ny tng cng sn xut cc ngoi c t gy sc. Bnh cnh lm sng
ca hi chng sc nhim c gm t ngt st cao, cm gic mt mi, tiu
chy ton nc, nhc u, au c, ni ban ngoi da v mt hoc nhiu du
hiu ca sc. Hi chng ny c t l t vong kh cao.
Hi chng Thukydides: l th c bit ca hi chng sc nhim c. Hi chng
ny c th gp thanh thiu nin c hai gii, thung gp do bi nhim t cu
sau khi b cm. T l t vong ca hi chng ny rt cao (trn 50%).

140

Ng c thc phm l bnh in hnh nht do c t rut bn vi nhit ca t

cu. Sau khi n cc thc n ngui hoc k c cc thc n nhim t cu c
nu chn. C th phn thnh hai th:
- Vim d dy rut: nguyn nhn do n phi cc thc n nhim c t t cu.
Bnh thng xy ra trong thi gian ngn vi triu chng nn ma d di, tiu
chy phn te nc, khng st. Bnh thng lnh nhanh chng v khng cn
iu tr khng sinh.
- Vim rut non-i trng: nguyn nhn do thc n nhim mt lng ln vi khun
t cu (>105 vi khun/g thc n) hoc do vi khun tng sinh trong lng rut
(c n 2-30% s ngi mang vi khun ny trong rut). Triu chng bnh l
ng tiu ha nh au bng, nn ma, tiu chy ko di hn v cn phi iu
tr khng sinh.
Nh vy, vi t l mu th hin hot tnh khng cc chng VSV kim nh
(c bit l khng khng 2 chng vi khun: S. aureus v P. aeruginosa) kh cao
ca cc mu sinh vt bin l mt tin quan trng phc v cho vic tm kim v
nghin cu cc hp cht thin nhin bin theo nh hng hot tnh khng sinh,
nhm pht trin cc ch phm thuc khng sinh th h mi, khc phc tnh trng
khng thuc khng sinh nghim trng hin nay.
Kt qu th hot tnh khng vi sinh vt kim nh ca cc phn on chit
phn b theo dung mi hu c ca cc mu c biu hin hot tnh cng cho thy
s ph hp vi cc ti liu c cng b. Chng hn nh, mu KC-018 (hi
min hnh chung Xesospongia testudinaria) th hin hot tnh khng 3 chng vi
sinh vt kim nh l P. aeruginosa, S. aureus v F. oxysporum. Kt qu nh gi
hot tnh ca cc phn on chit phn b bng dung mi hu c cho thy, hot
tnh tp trung ch yu phn on clorofooc v hexan, phn on butanol v
nc khng biu hin hot tnh. Theo ti liu cng b, thnh phn ha hc ch
yu v c hot tnh khng sinh ca chi ny l cc hp cht axit mch di, a ni
i v b brm ha. Cc hp cht ny u c phn cc rt thp nn phn b
ch yu cc phn on hexan v clorofooc, to nn hot tnh khng sinh ca
cc phn on ny. Ngc li, hot tnh khng sinh ca cc mu hi sm nh KC029, KC-057 v KC-061 li nm ch yu cc phn on butanol v nc. iu
ny c th c l gii bi thnh phn ha hc chnh ca chi Holothuria l cc
hp cht holothurin nhn t chnh to nn hot tnh, u l cc hp cht glycosit
c phn cc rt ln nn phn b cc phn on butanol v nc, to nn s
tp trung hot tnh cc phn on ny. Cc kt qu ny s nh hng cho cc
nghin cu tip theo.
II.3. Kt qu nh gi hot tnh gy c t bo
(cytotoxic activity)
Gi tr CS% hay chnh l gi tr biu hin lng t bo sng st sau khi
cho tip xc vi cc mu chit c tnh theo cc cng thc trnh by trong
phn quy trnh. Kt qu xc nh gi tr CS% ca 101 mu sinh vt bin c
trnh by trn bng 4.

141

 y cc mu c gi tr t bo sng st cng nh chng t hot tnh cng

cao. Tuy nhin, theo tiu chun nh gi ca Vin nghin cu Ung th M (NCI)
th cc mu th nghim sng lc s cp c gi tr CS% <50 s c coi l c hot
tnh v chun b cho bc th tip theo tm gi tr c ch ti 50% s pht trin
ca cc dng t bo th. Cc mu c biu hin hot tnh l cc mu c gi tr IC50
< 20 g/ml.
Bng 4: Bng tng kt kt qu th hot tnh gy c t bo ca 101 mu sinh vt
bin
K hiu mu
Chng (-)
Chng (+)

Gi tr CS% (Cell survival)

KB

FL

Hep-G2

1000,0

1000,0

100,00,0

Ghi ch

1,071,07

1,520,00

1,340,63

KC-001

44,51,5

40,12,2

33,71,8

Khng c 3 dng t bo

KC-002

47,32,1

Khng 2 dng t bo: FL v Hep-G2

57,61,9

34,51,7

KC-003

60,52,5

77,13,7

66,82,3

m tnh

KC-004

46,71,8

51,91,2

44,12,0

Khng 2 dng t bo: KB v Hep-G2

KC-005

20,11,3

47,82,1

32,71,9

Khng c 3 dng t bo

KC-006

70,52,7

67,81,9

89,02,1

m tnh

KC-007

66,71,5

30,23,4

37,83,1

Khng 2 dng t bo: FL v Hep-G2

KC-008

45,61,4

65,34,5

54,13,9

Khng mt dng t bo: KB

KC-009

78,93,2

66,72,1

43,62,7

Khng 1 dng t bo:Hep-G2

KC-010

62,11,9

77,52,1

69,84,7

m tnh

KC-011

83,72,3

55,41,7

79,23,1

m tnh

KC-012

65,73,4

78,15,6

66,72,3

m tnh

KC-013

79,62,1

74,93,4

56,70,8

m tnh

KC-014

73,72,3

85,42,7

89,23,5

m tnh

KC-015

55,72,4

68,11,6

56,74,3

m tnh

KC-016

63,73,3

55,41,2

79,22,1

m tnh

KC-017

56,71,6

44,32,5

37,61,1

Khng 2 dng t bo: FL v Hep-G2

KC-018

73,73,3

85,41,7

69,22,6

m tnh

KC-019

71,52,3

75,41,9

74,22,1

m tnh

KC-020

30,13,3

47,82,1

42,71,3

Khng c 3 dng t bo

KC-021

81,71,3

59,41,7

89,23,1

m tnh

KC-022

77,82.2

44,72,5

37,41,9

Khng 2 dng t bo: FL v Hep-G2

KC-023

42,12.4

67,81,5

40,92,7

Khng 2 dng t bo: KB v Hep-G2

KC-024

63,72,3

75,44,7

79,22,1

m tnh

KC-025

40,81,9

32,53,5

37,81,8

Khng c 3 dng t bo

KC-026

93,72,3

75,44,7

89,23,5

m tnh

KC-027

68,73,3

57,41,2

69,22,1

m tnh

142

KC-028

75,73,1

65,41,9

69,52,6

m tnh

KC-029

30,82,9

35,52,5

27,81,2

Khng c 3 dng t bo

KC-030

58,31,8

44,12,3

37,91,9

Khng 2 dng t bo: FL v Hep-G2

KC-031

63,71,3

75,42,7

89,23,5

m tnh

KC-032

70,22,2

68,61,8

44,72,5

Khng 1 dng t bo: Hep-G2

KC-033

Khng 2 dng t bo: FL v KB

34,71,5

42,12,2

66.83.9

KC-034

55,73,1

71,61,9

59,52,6

m tnh

KC-035

48,21,5

55,72,1

56,83,0

Khng 1 dng t bo: KB

KC-036

83,71,9

65,44,7

79,21,5

m tnh

KC-037

33,71,7

47,02,1

58,91,5

Khng 2 dng t bo: FL v KB

KC-038

53,71,9

75,42,7

59,24,5

m tnh

KC-039

40,83,9

45,51,5

37,82,2

Khng c 3 dng t bo

KC-040

63,71,9

75,43,7

69,22,5

m tnh

KC-041

59,72,1

64,41,7

89,53,3

m tnh

KC-042

55,62,7

43,21,3

46,71,6

Khng 2 dng t bo: FL v Hep-G2

KC-043

32,81,9

40,52,1

37,81,2

Khng c 3 dng t bo

KC-044

22,53,4

27,21,9

35,42,0

Khng c 3 dng t bo

KC-045

44,71,5

66,81,1

71,91,8

Khng 1 dng t bo: KB

KC-046

73,71,9

85,43,7

59,22,5

m tnh

KC-047

63,62,3

65,21,6

57,53,1

m tnh

KC-048

55,61,3

48,62,1

69,71,4

Khng 1 dng t bo: FL

KC-049

59,52,1

64,61,7

91,53,3

m tnh

KC-050

80,22,3

54,71,9

49,60,8

Khng 1 dng t bo: Hep-G2

KC-051

33,22,3

55,94,3

47,12,0

Khng 2 dng t bo: KB v Hep-G2

KC-052

44,71,8

51,91,2

39,12,0

Khng 2 dng t bo: KB v Hep-G2

KC-053

75,73,3

61,43,2

62,22,1

m tnh

KC-054

58,72,3

65,42,9

69,51,8

m tnh

KC-055

68,71,7

713,1

56,63,2

m tnh

KC-056

32,72,4

41,81,2

76,32,3

Khng 2 dng t bo: FL v KB

KC-057

43,81,9

39,52,1

37,81,5

Khng c 3 dng t bo

KC-058

59,52,1

64,61,7

49,81,5

m tnh

KC-059

53,71,9

65,43,1

59,22,2

m tnh

KC-060

42,52,4

27,91,6

35,43,0

Khng c 3 dng t bo

KC-061

32,51,9

37,93,1

25,92,5

Khng c 3 dng t bo

KC-062

68,71,3

75,41,2

79,21,1

m tnh

KC-063

73,73,3

65,43,2

69,22,1

m tnh

KC-064

53,71,9

65,44,7

79,21,5

m tnh

KC-065

66,71,5

35,22,4

37,23,1

Khng 2 dng t bo: FL v Hep-G2

KC-066

63,51,3

75,42,5

79,23,2

m tnh

KC-067

73,52,7

55,43,1

69,21,5

m tnh

KC-068

48,71,2

57,83,5

59,91,7

Khng 1 dng t bo: KB

143

KC-069

68,21,3

75,71,2

78,21,1

m tnh

KC-070

73,62,3

65,45,2

37,93,1

Khng 1 dng t bo: Hep-G2

KC-071

57,21,9

58,44,1

46,71,6

Khng 1 dng t bo: Hep-G2

KC-072

83,71,3

65,41,7

79,21,4

m tnh

KC-073

57,71,9

85,44,8

79,11,5

m tnh

KC-074

63,91,9

63,43,6

89,31,5

m tnh

KC-075

46,82,6

40,52,1

63,62,3

Khng 2 dng t bo: KB v FL

KC-076

58,72,3

65,42,9

69,51,8

m tnh

KC-077

80,22,3

54,71,9

43,61,8

Khng 1 dng t bo: Hep-G2

KC-078

63,72,3

75,44,7

79,22,1

m tnh

KC-079

55,72,4

68,11,6

56,74,3

m tnh

KC-080

33,82,9

43,52,1

37,81,5

Khng c 3 dng t bo

KC-081

40,83,9

45,51,5

47,22,2

Khng c 3 dng t bo

KC-082

83,72,3

55,41,7

79,23,1

m tnh

KC-083

58,72,3

65,42,9

69,51,8

m tnh

KC-084

68,22,1

85,72,1

59,52,8

m tnh

KC-085

59,72,1

64,41,7

89,53,3

m tnh

KC-086

42,81,9

40,52,1

47,81,2

Khng c 3 dng t bo

KC-087

55,61,3

44,62,1

69,71,4

Khng 1 dng t bo: FL

KC-088

78,71,7

71,73,1

56,63,2

m tnh

KC-089

33,81,9

43,52,1

37,81,5

Khng c 3 dng t bo

KC-090

78,73,3

57,41,2

69,22,1

m tnh

KC-091

83,72,3

75,44,7

83,23,5

m tnh

KC-092

68,73,3

57,41,2

69,22,1

m tnh

KC-093

85,73,1

65,51,9

64,52,6

m tnh

KC-094

53,51,3

85,42,5

79,23,2

m tnh

KC-095

72,11,9

77,52,1

69,82,7

m tnh

KC-096

83,72,3

65,41,7

79,23,1

m tnh

KC-097

85,73,4

78,15,6

66,72,3

m tnh

KC-098

79,62,1

74,93,4

56,50,8

m tnh

KC-099

73,72,3

85,42,7

89,23,5

m tnh

KC-100

55,72,4

68,11,6

56,74,3

m tnh

KC-101

43,21,3

75,94,3

37,12,0

Khng 2 dng t bo: KB v Hep-G2

Kt qu nghin cu cho thy c 43 mu c biu hin hot tnh trong tng

s 101 mu th, trong c 12 mu khng mt dng t bo (chim 11,88 %), 15
mu khng 2 dng t bo (chim 14,85 %) v 16 mu khng c 3 dng t bo
(chim 15,84 %). Kt qu c th hin trn bng 5.

144

Bng 5. Bng tng kt kt qu sng lc hot tnh gy c t bo ca 101 mu

Tng s mu

S mu c hot C hot tnh vi C hot tnh vi C hot tnh vi

tnh
1 dng t bo 2 dng t bo
3 dng t bo

S mu

S mu

S mu

S mu

S mu

101

100

43

42,57

12

11,88

15

14,85

16

15,84

Trong s 101 mu, c 30 mu th hin hot tnh khng dng t bo KB

(ung th biu m), 28 mu khng dng t bo FL (ung th mng t cung) v 33
mu khng dng t bo Hep-G2 (ung th gan). Kt qu c tng hp trn bng
6.
Bng 6: Bng tng kt kh nng cc dng KB ,FL v Hep-G2ca 101 mu
C hot tnh
khng KB

C hot tnh
khng FL

C hot tnh khng

Hep-G2

S mu c hot tnh

30

28

33

T l % trn 101 mu th

29,70

27,72

32,67

15.84

34
32.67

33
4.85

32

11.88

T l %

31
42.57

30

29.7

29
27.72

28
Mu c hot tnh

Mu khng 1 dng TB

Mu khng 2 dng TB

Mu khng 3 dng TB

27
26
25
Khng dng t bo
KB

Khng dng t bo
FL

Khng dng t bo
Hep-G2

Biu 3:Biu so snh hot tnh gy c t bo ca cc mu sinh vt bin

T kt qu trn, cc mu c biu hin hot tnh gy c t bo c tin
hnh xc nh gi tr IC50 (nng c ch 50% s pht trin ca t bo). ng
thi, cc mu ny cng c tin hnh chit phn b trong cc dung mi hu c
v cc phn on thu c cng c tin hnh xc nh gi tr IC50. Kt qu th
nghim c tng hp trong bng 7.

145

Bng 7: Tng hp kt qu xc nh IC50 ca cc mu sinh vt bin c biu hin

hot tnh v cc dch chit phn on ca chng.
K hiu mu

KC-001

KC-002

KC-004

KC-005

KC-007

KC-008

KC-009

KC-017

Dch
chit

Gi tr IC50 (g/ml)

Ghi ch

KB

FL

Hep-G2

17,5

18,3

12,8

(-)

(-)

(-)

16,7

18,1

(-)

Khng 2 dng

(-)

19,0

10,4

Khng 2 dng

(-)

(-)

(-)

(-)

16,5

19,3

Khng 2 dng

(-)

19,5

(-)

Khng 1 dng

(-)

15,7

18,6

Khng 2 dng

(-)

(-)

(-)

Khng c 3 dng

(-)

(-)

(-)

17,4

(-)

16,9

(-)

(-)

(-)

18,0

(-)

15,7

Khng 2 dng

19,9

(-)

16,0

Khng 2 dng

(-)

(-)

(-)

10,3

18,7

15,6

Khng c 3 dng

9,7

14,3

13,9

Khng c 3 dng

11,5

17,9

18,6

Khng c 3 dng

(-)

(-)

(-)

(-)

(-)

(-)

(-)

16,5

17,2

(-)

(-)

(-)

(-)

12,7

18,6

Khng 2 dng

Khng 2 dng

Khng 2 dng

Khng 2 dng

(-)

17,0

15,3

(-)

(-)

(-)

18,4

(-)

(-)

Khng 1 dng

15,6

(-)

(-)

Khng 1 dng

12.1

(-)

(-)

Khng 1 dng

(-)

(-)

(-)

(-)

(-)

(-)

(-)

(-)

17,6

Khng 1 dng

(-)

(-)

14,5

Khng 1 dng

(-)

(-)

15,3

Khng 1 dng

(-)

(-)

19,9

Khng 1 dng

(-)

(-)

(-)

(-)

18,5

16,1

146

Khng 2 dng

KC-020

KC-022

KC-023

KC-025

KC-029

KC-030

KC-032

KC-033

(-)

(-)

19,8

Khng 1 dng

(-)

16,3

15,8

Khng 2 dng

(-)

(-)

(-)

(-)

(-)

(-)

15,6

18,9

16,8

Khng c 3 dng

14,2

15,4

13,1

Khng c 3 dng

18,7

17,6

19,8

Khng c 3 dng

(-)

(-)

(-)

(-)

(-)

(-)

(-)

16,5

14,6

(-)

(-)

(-)

(-)

15,4

13,7

(-)

(-)

(-)

(-)

19,9

(-)

Khng 1 dng

16,7

(-)

14,9

Khng 2 dng

15,5

(-)

14,0

Khng 2 dng

16,0

(-)

15,1

Khng 2 dng

(-)

(-)

(-)

(-)

(-)

(-)

14,3

13,7

14,0

Khng c 3 dng

13,5

13,0

12,7

Khng c 3 dng

15,1

14,0

12,9

Khng c 3 dng

(-)

(-)

(-)

Khng 2 dng
Khng 2 dng

(-)

(-)

(-)

14,8

16,7

12,3

(-)

(-)

(-)

(-)

(-)

(-)

14,1

15,2

13,0

Khng c 3 dng

18,9

(-)

16,7

Khng 2 dng

(-)

17,6

18,2

Khng 2 dng

(-)

(-)

17,9

Khng 1 dng

(-)

15,4

16,5

Khng 2 dng

(-)

(-)

(-)

(-)

(-)

(-)

(-)

(-)

17,3

(-)

(-)

(-)

(-)

(-)

15,6

Khng 1 dng

(-)

(-)

18,7

Khng 1 dng

(-)

(-)

(-)

14,6

18,1

(-)

Khng 2 dng

(-)

17,5

(-)

Khng 1 dng

147

Khng c 3 dng

Khng 1 dng

KC-035

KC-037

KC-039

KC-042

KC-043

KC-044

KC-045

KC-048

13,5

15,7

(-)

(-)

(-)

(-)

Khng 2 dng

(-)

(-)

(-)

18.9

(-)

(-)

(-)

(-)

(-)

15,4

(-)

(-)

(-)

(-)

(-)

(-)

(-)

(-)

13,4

18,9

(-)

(-)

(-)

(-)

15,7

16,5

(-)

Khng 2 dng

(-)

15,4

(-)

Khng 1 dng

Khng 1 dng

Khng 2 dng

(-)

(-)

(-)

16,7

17,8

15,2

Khng c 3 dng

14,3

12,1

15,0

Khng c 3 dng

15,1

14,8

(-)

Khng 2 dng

(-)

(-)

19,9

Khng 1 dng

(-)

(-)

(-)

(-)

16,2

18,5

(-)

(-)

(-)

(-)

15,6

17,7

Khng 2 dng

(-)

16,1

14,3

Khng 2 dng

(-)

(-)

(-)

14,2

18,6

17,9

(-)

(-)

(-)

12,1

15,7

14,3

(-)

(-)

(-)

(-)

(-)

(-)

12,7

14,3

17,2

Khng c 3 dng

11,9

12,6

16,9

Khng c 3 dng

15,0

13,2

18,6

Khng c 3 dng

(-)

(-)

(-)

(-)

(-)

(-)

Khng 2 dng

Khng c 3 dng
Khng c 3 dng

18,6

(-)

(-)

(-)

(-)

(-)

17,5

(-)

(-)

Khng 1 dng
Khng 1 dng

19,8

(-)

(-)

(-)

(-)

(-)

(-)

17,9

(-)

(-)

(-)

(-)

(-)

19,8

(-)

148

Khng 1 dng

Khng 1 dng
Khng 1 dng

KC-050

KC-051

KC-052

KC-056

KC-057

KC-060

KC-061

KC-065

(-)

16,7

(-)

Khng 1 dng

(-)

19,5

(-)

Khng 1 dng

(-)

(-)

19,3

Khng 1 dng

(-)

(-)

19,7

Khng 1 dng

(-)

(-)

17,3

Khng 1 dng

15,6

Khng 1 dng

(-)

(-)

(-)

(-)

(-)

14,7

(-)

18,5

17,9

(-)

(-)

15,3

(-)

14,7

(-)

(-)

(-)

(-)

(-)

(-)

18,9

(-)

16,5

Khng 2 dng

(-)

(-)

19,8

Khng 1 dng

14,3

(-)

(-)

Khng 1 dng

18,2

(-)

17,9

Khng 2 dng

(-)

(-)

(-)

17,6

18,1

(-)

(-)

(-)

(-)

(-)

19,5

(-)

Khng 1 dng

15,1

14,9

(-)

Khng 2 dng

18,7

(-)

(-)

15,2

Khng 2 dng
Khng 2 dng

Khng 2 dng

17,9

16,0

(-)

(-)

(-)

16,9

(-)

18,6

Khng 2 dng

15,7

14,3

12,1

Khng c 3 dng

(-)

19,5

(-)

17,3

14,1

16,8

Khng c 3 dng

15,6

13,0

14,7

Khng c 3 dng

13,1

17,9

(-)

Khng 2 dng

Khng c 3 dng

(-)

(-)

(-)

(-)

(-)

(-)

15,7

16,5

12,1

(-)

(-)

(-)

19,8

(-)

16,5

Khng 2 dng

12,1

13,5

10,9

Khng c 3 dng

Khng c 3 dng

17,6

(-)

18,1

Khng 1 dng

(-)

16,7

18,3

Khng 2 dng

(-)

(-)

(-)

(-)

12,6

11,7

Khng 2 dng

(-)

15,5

16,8

Khng 2 dng

149

KC-068

KC-070

KC-071

KC-075

KC-077

KC-080

KC-081

KC-086

(-)

(-)

(-)

18,5

(-)

(-)

(-)

(-)

(-)

16,7

(-)

(-)

Khng 1 dng

17,1

(-)

(-)

Khng 1 dng

(-)

(-)

(-)

(-)

(-)

15,6

(-)

(-)

(-)

(-)

(-)

19,4

Khng 1 dng

(-)

(-)

15,3

Khng 1 dng

(-)

(-)

17,2

Khng 1 dng

(-)

(-)

18,5

Khng 1 dng

(-)

(-)

(-)

(-)

(-)

17,3

Khng 1 dng

(-)

(-)

18,6

Khng 1 dng

(-)

(-)

(-)

18,6

15,1

(-)

Khng 2 dng

15,4

12,9

(-)

Khng 2 dng

16,7

(-)

(-)

Khng 1 dng

Khng 1 dng

Khng 1 dng

(-)

(-)

(-)

(-)

(-)

(-)

(-)

(-)

15,8

Khng 1 dng

(-)

(-)

12,1

Khng 1 dng

(-)

(-)

18,6

Khng 1 dng

(-)

(-)

(-)

(-)

(-)

(-)

16,3

15,1

17,8

Khng c 3 dng

15,6

13,0

14,7

Khng c 3 dng

13,1

17,9

(-)

Khng 2 dng

(-)

(-)

(-)

(-)

(-)

(-)

17,7

11,5

12,1

(-)

(-)

(-)

19,8

(-)

16,5

Khng 2 dng

12,1

9,5

10,9

Khng c 3 dng

17,6

(-)

18,1

Khng 2 dng

15,5

16,7

18,3

Khng c 3 dng

(-)

(-)

(-)

14,4

12,6

11,7

(-)

(-)

(-)

(-)

(-)

(-)

150

Khng c 3 dng

Khng c 3 dng

KC-087

KC-089

KC-101

(-)

17,8

(-)

(-)

(-)

(-)

(-)

(-)

(-)

(-)

15,5

(-)

(-)

(-)

(-)

16,7

14,5

17,6

(-)

(-)

(-)

(-)

(-)

(-)

Khng 1 dng

Khng 1 dng
Khng c 3 dng

12,1

9,5

10,9

Khng c 3 dng

17,6

17,8

18,1

Khng 3 dng

18,2

(-)

17,5

Khng 2 dng

(-)

(-)

(-)

(-)

(-)

15,6

Khng 1 dng

15,7

(-)

13,4

Khng 2 dng

19,8

(-)

(-)

Khng 1 dng

II.4. Kt qu nh gi hot tnh chng xy ho ca cc

mu sinh vt bin thng qua phn ng bao vy gc t do
DPPH (DPPH free radical scanvenging activity)
DPPH c kh nng to ra cc gc t do bn trong dung dch EtOH bo
ho. Khi cho cc cht th nghim vo hn hp ny, nu cht c kh nng lm
trung ho hoc bao vy cc gc t do s lm gim cng hp ph nh sng ca
cc gc t do DPPH. Hot tnh chng xy ho c nh gi thng qua gi tr
hp ph nh sng ca dch th nghim so vi i chng khi c trn my Elisa
bc sng 515nm.
Gi tr SC biu hin kh nng bt gi cc gc t do thng qua h xy ho
t to bi DPPH, c tnh theo cng thc c trnh by trong quy trnh trn.
Kt qu th nghim v tnh ton gi tr SC (%) ca 101 mu sinh vt bin c
trnh by trong bng 8.
Bng 8:Kt qu gi tr t l gc t do b bao vy SC% ca cc mu SVB
K hiu mu

Gi tr SC%

Ghi ch

Chng (-)

0,000,00

m tnh

Chng (+)

81,220,52

dng tnh

KC-001

52,101,65

dng tnh

KC-002

56,542,02

dng tnh

KC-003

30,441,52

m tnh

KC-004

42,151,67

m tnh

KC-005

58,652,55

dng tnh

KC-006

25,462,34

m tnh

151

KC-007

54,721,18

dng tnh

KC-008

37,771,32

m tnh

KC-009

35,682,31

m tnh

KC-010

50,891,12

dng tnh

KC-011

15,342,11

m tnh

KC-012

46,221,11

m tnh

KC-013

22,451,33

m tnh

KC-014

57,151,75

dng tnh

KC-015

42,181,23

m tnh

KC-016

29,112,10

m tnh

KC-017

71,451,33

dng tnh

KC-018

37,983,01

m tnh

KC-019

44,672,53

m tnh

KC-020

45,123,44

m tnh

KC-021

16,551,23

m tnh

KC-022

34,172,05

m tnh

KC-023

35,991,67

m tnh

KC-024

47,331,11

m tnh

KC-025

73,211,87

dng tnh

KC-026

19,211,64

m tnh

KC-027

42,681,46

m tnh

KC-028

40,992,17

m tnh

KC-029

33,691,98

m tnh

KC-030

30,551,35

m tnh

KC-031

46,671,23

m tnh

KC-032

21,341,88

m tnh

KC-033

23,113,01

m tnh

KC-034

44,332,33

m tnh

KC-035

18,961,01

m tnh

KC-036

47,221,22

m tnh

KC-037

11,683,54

m tnh

KC-038

40,201,24

m tnh

KC-039

33,772,11

m tnh

KC-040

43,231,89

m tnh

KC-041

22,771,99

m tnh

KC-042

53,131,62

dng tnh

KC-043

16,552,33

m tnh

KC-044

59,011,77

dng tnh

KC-045

41,981,22

m tnh

KC-046

36,792,10

m tnh

KC-047

17,831,25

m tnh

152

KC-048

47,111,54

m tnh

KC-049

57,641,51

dng tnh

KC-050

57,102,23

dng tnh

KC-051

33,552,87

m tnh

KC-052

54,231,60

dng tnh

KC-053

21,992,33

m tnh

KC-054

34,671,78

m tnh

KC-055

42,192,15

m tnh

KC-056

60,231,37

dng tnh

KC-057

31,792,33

m tnh

KC-058

55,951,45

dng tnh

KC-059

37,861,67

m tnh

KC-060

58,752,12

dng tnh

KC-061

71,771,11

dng tnh

KC-062

28,672,11

m tnh

KC-063

44,691,00

m tnh

KC-064

47,230,98

m tnh

KC-065

19,641,43

m tnh

KC-066

39,601,68

m tnh

KC-067

46,112,35

m tnh

KC-068

39,671,47

m tnh

KC-069

29,813,67

m tnh

KC-070

12,221,69

m tnh

KC-071

43,571,86

m tnh

KC-072

52,151,96

dng tnh

KC-073

40,992,17

m tnh

KC-074

33,691,98

m tnh

KC-075

40,551,35

m tnh

KC-076

46,671,23

m tnh

KC-077

31,341,88

m tnh

KC-078

23,113,01

m tnh

KC-079

24,332,33

m tnh

KC-080

18,961,01

m tnh

KC-081

68,731,56

dng tnh

KC-082

11,683,54

m tnh

KC-083

21,992,33

m tnh

KC-084

34,671,78

m tnh

KC-085

42,192,15

m tnh

KC-086

34,671,78

m tnh

KC-087

42,192,15

m tnh

KC-088

65,551,05

dng tnh

153

KC-089

71,651,12

dng tnh

KC-090

46,221,11

m tnh

KC-091

22,451,33

m tnh

KC-092

37,983,01

m tnh

KC-093

44,672,53

m tnh

KC-094

45,123,44

m tnh

KC-095

16,551,23

m tnh

KC-096

55,482,24

dng tnh

KC-097

18,961,01

m tnh

KC-098

47,221,22

m tnh

KC-099

11,683,54

m tnh

KC-100

40,201,24

m tnh

KC-101

72,551,32

dng tnh

Kt qu xc nh gi tr SC cho thy, c 23/101 mu (chim t l 22,72 %)

c biu hin dng tnh vi gi tr SC > 50%. Trong , c 18 mu c kh nng
bao vy t 50-70% cc gc t do trong th nghim DPPH v 5 mu c kh nng
bao gy trn 70% cc gc t do. Kt qu tng hp c th hin trn bng 8 v
biu 4.
Bng 8: Tng hp kt qu hot tnh chng xy ho ca cc mu sinh vt bin
Tng s
mu

S mu c
hot tnh

S mu T l %
101
100

S mu
23

T l %
22,72

C gi tr SC
t 50-70%
S mu
18

T l %
17,82

C gi tr
SC >70%
S mu
5

T l %
4,95

120
100

100
77.28

T l %

80
60
40
22.72

20

17.82
4.95

0
Mu dng Mu c SC Mu c SC
tnh
t 50-70%
> 70%

Mu m
tnh

Tng s
mu

Biu 4. So snh t l % cc mu th hin hot tnh chng OXH

154

Bng 9. Tng hp gi tr SC% v SC50 ca cc mu sinh vt bin c biu hin

dng tnh v cc phn on chit ca chng.
K hiu mu

Dch chit

Chng (-)
Chng (+)

KC-001

KC-002

KC-005

KC-007

KC-010

KC-014

KC-017

Gi tr SC%

Gi tr SC50(g/ml)

Ghi ch

0,000,00

>200

m tnh

81,220,52

20,1

dng tnh

52,101,65

77,6

dng tnh

33,471,28

(-)

m tnh

46,232,01

(-)

m tnh

59,131,56

68,5

dng tnh

18,241,67

(-)

m tnh

56,542,02

70,4

dng tnh

50,981,00

76,7

dng tnh

57,891,23

70,1

dng tnh

44,211,78

(-)

m tnh

34,551,75

(-)

m tnh

58,652,55

68,3

dng tnh

60,771,44

65,9

dng tnh

58,681,33

69,3

dng tnh

42,331,45

(-)

m tnh

39,561,34

(-)

m tnh

54,721,18

75,1

dng tnh

34,551,67

(-)

m tnh

55,771,83

71,5

dng tnh

60,932,86

64,8

dng tnh

44,211,11

(-)

m tnh

50,891,12

80,1

dng tnh

59,881,23

66,1

dng tnh

55,572,55

71,0

dng tnh

33,441,22

(-)

m tnh

21,351,78

(-)

m tnh

57,151,75

60,5

dng tnh

62,571,55

59,7

dng tnh

57,651,22

61,5

dng tnh

44,331,89

(-)

m tnh

37,862,22

(-)

m tnh

71,451,33

30,2

dng tnh

29,342,21

(-)

m tnh

73,551,55

27,5

dng tnh

57,552,11

49,6

dng tnh

34,561,45

(-)

m tnh

155

KC-025

KC-042

KC-044

KC-049

KC-050

KC-052

KC-056

KC-058

KC-060

73,211,87

45,9

dng tnh

80,231,22

29,7

dng tnh

76,561,77

41,0

dng tnh

56,631,34

55,6

dng tnh

34,563,21

(-)

m tnh

53,131,62

66,8

dng tnh

48,111,45

(-)

m tnh

55,641,37

62,4

dng tnh

23,671,82

(-)

m tnh

22,461,89

(-)

m tnh

59,011,77

62,3

dng tnh

60,111,66

53,1

dng tnh

55,641,77

67,9

dng tnh

43,672,22

(-)

m tnh

32,461,89

(-)

m tnh

57,641,51

65,6

dng tnh

34,562,11

(-)

m tnh

60,341,99

64,2

dng tnh

45,671,88

(-)

m tnh

43,213,65

(-)

m tnh

57,102,23

68,2

dng tnh

52,661,51

72,1

dng tnh

57,791,34

60,9

dng tnh

45,211,78

(-)

m tnh

34,651,75

(-)

m tnh

54,231,60

69,7

dng tnh

50,980,76

75,8

dng tnh

55,881,67

70,0

dng tnh

23,771,44

(-)

m tnh

22,462,11

(-)

m tnh

60,231,37

55,6

dng tnh

22,131,66

(-)

m tnh

51,441,78

79,1

dng tnh

65,472,15

63,9

dng tnh

52,671,44

81,2

dng tnh

55,951,45

67,3

dng tnh

52,121,68

75,3

dng tnh

58,341,57

68,9

dng tnh

34,211,81

(-)

m tnh

42,552,11

(-)

m tnh

58,752,12

52,4

dng tnh

156

KC-061

KC-072

KC-081

KC-088

KC-089

KC-096

KC-101

61,55167

50,9

dng tnh

57,701,86

57,1

dng tnh

42,831,66

(-)

m tnh

38,731,46

(-)

m tnh

71,771,11

46,6

dng tnh

34,572,11

(-)

m tnh

55,681,69

82,6

dng tnh

75,692,33

42,5

dng tnh

73,541,78

50,6

dng tnh

52,151,96

88,5

dng tnh

55,572,11

85,6

dng tnh

60,631,29

82,6

dng tnh

45,721,32

(-)

m tnh

33,651,11

(-)

m tnh

71,771,11

46,6

dng tnh

34,572,11

(-)

m tnh

54,881,25

83,2

dng tnh

65,692,12

63,5

dng tnh

44,371,34

(-)

m tnh

65,551,05

60,4

dng tnh

50,981,00

76,7

dng tnh

77,891,23

50,1

dng tnh

43,211,78

(-)

m tnh

35,551,75

(-)

m tnh

60,531,22

65,6

dng tnh

22,131,66

(-)

m tnh

41,441,78

(-)

m tnh

65,472,15

63,7

dng tnh

52,671,44

75,9

dng tnh

55,482,29

65,7

dng tnh

34,562,11

(-)

m tnh

60,341,54

64,1

dng tnh

45,671,88

(-)

m tnh

43,213,65

(-)

m tnh

72,721,32

31,2

dng tnh

29,342,21

(-)

m tnh

32,131,62

(-)

m tnh

73,851,55

26,6

dng tnh

N
34,561,45
(-)
m tnh
Ch thch: M - Cn dch tng metanol, H - Cn chit hexan, C - Cn chit clorofooc, B - Cn chit
butanol, N - Cn chit nc.

157

Cc mu c biu hin dng tnh (SC% > 50 %) c tip tc tin hnh

xc nh gi tr SC50 (gi tr nng mu th c kh nng bao vy 50% cc gc
t do), ng thi tin hnh chit phn b bng cc dung mi hu c. Cc phn
on chit ny c tip tc tin hnh xc nh gi tr SC%. Cc phn on c
biu hin dng tnh li tip tc c xc nh gi tr SC50. Kt qu th nghim
c tng hp trong bng 9. Kt qu trn cho thy s ph hp so vi cc ti liu
c cng b. Chng hn vi cc mu KC-007 (Cladiella sp), KC-010
(Lobophytum sp), KC-014 (Sinularia abrupta), KC-025 (Sarcophyton
mililatensis) c biu hin hot tnh c th c gii thch bi thnh phn chnh
ca cc chi san h mm ny l cc hp cht itecpen dng cembranoit th hin
nhiu hot tnh sinh hc ng quan tm trong c c hot tnh chng xy ha
[162] - [165]. y l lp cht c phn cc thp nn s phn b ch yu dch
chit hexan v cloroform, gii thch cho s tp trung hot tnh vo cc dch chit
ny ca cc mu san h mm trn. Cn cc mu KC-056 (Astropecten
polyacanthus) v KC-061 (Holothuria scabra) c hot tnh tp trung cc phn
on BuOH v nc, c th c gii thch bi thnh phn hot cht chnh ca
cc chi sao bin v hi sm trn u l cc steroit glycosit cha nhiu n v
ng trong phn t, c phn cc rt cao v do phn b ch yu cc phn
on BuOH v nc [166] - [167].

158

B. nghin cu ha hc theo nh hng hot

tnh sinh hc mt s i tng c la chn
Trn c s cc kt qu nghin cu thu thp, gim nh loi, to dch chit
v sng lc hot tnh sinh hc, kt hp vi cc tiu ch la chn l:
- Hot tnh sinh hc
- Mc kh khn trong cng tc ly mu
- Tr lng thc t ca loi v kh nng c th thu thp mu
- Tra cu cc nghin cu v hot tnh sinh hc v thnh phn ha
hc cng b v loi hoc chi ca i tng nghin cu.
07 i tng ph hp nht c la chn tin hnh thu thp mu
lng ln, to dch chit phc v cho nghin cu ha hc l:
Tn Vit Nam

Tn khoa hc

San h mm

Sarcophyton mililatensis

San h mm

Cladiella sp.

Hi sm trng

Holothuria scabra

Sao bin

Archaster typicus

Hi min xp

Xestospongia testudinaria

Hi min cnh Gellius varius

xanh
Hi min
Ianthella sp.

a im thu
mu
Alcyoniidae
Cn o, B
Ra - Vng tu
Alcyoniidae
Cn C,
Qung Tr
Holothuridae Ct B, Hi
Phng
Archasteridae Ct B, Hi
phng
Petrosiidae
Qun
o
Trng
Sa,
Khnh Ha
Chanlinidae
H
Long,
Qung Ninh
Ianthellidae
Qun
o
Trng
Sa,
Khnh Ha
H

Kt qu nghin cu chi tit v thnh phn ha hc ca tng i tng theo

nh hng hot tnh sinh hc (bioassay guided) c trnh by di y.

159

Chng III. Nghin cu ho hc theo nh hng

hot tnh sinh hc ca mt s loi san h mm
III.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca
loi san h mm sarcophyton mililatensis (ALcyoniidae)
III.1.1. M u
Mu san h mm Sarcophyton mililatensis c thu thp ti hn By
Cnh, huyn o Cn o, tnh B Ra - Vng Tu vo thng 6 nm 2007 v
c TS. Cng Thung v cc cng s, Vin Ti nguyn v Mi trng Bin,
Vin KH&CN Vit Nam gim nh tn khoa hc.
- c im: Tp on c
thn ln, polyparium c
dng lng cho, pha ngoi
gn sng hoc gp khc. B
mt tp on nhm, polyp
nh. mi trng sng
mnh thn ngn v mp,
mi trng c bo v thn
di v mnh hn
- Mu sc: c mu vng
xanh ln nu hoc nu vng

San h mm: Sarcophyton mililatensis

San h l ng vt bin c to thnh bi nhng sinh vt rt nh gi l
cc polyp san h. Mi polyp san h ging nh mt cy to bin vi thn dng ti
v ch c mt ming m ly thc n v loi cht thi. Xung quanh ming ny
l cc xc tu vi cc t bo gy nga. Mt tp on san h khng phi l mt
nhm cc polyp n l cng sng vi li ch chung, m l kt qu ca s trng
thnh v m chi ca mt polyp c s. Cc polyp trong mt cm san h c
chung mt h cht lng v thn kinh. Tt c u ging nhau v gen v cc polyp
lin kt vi nhau bi mt lp m mng. Cc rn san h hnh thnh trn cc b
mc vng chc nhng vng bin m, nng, v nc trong. Cc rn san h l
mt trong nhng h sinh thi lu i nht, a dng nht v mt sinh hc, v
phong ph v cc loi trn tri t. Chng cn h tr mi quan h cng sinh
(cng c li) gia cc sinh vt trong th gii t nhin. Mc d ch do nhng sinh
vt rt nh to thnh, san h to nn nhng rn tuyt p dc theo b bin Vit
Nam v l ni sinh sng ca nhiu loi sinh vt bin a dng.
San h rt nhy cm vi s xo trn, v s tn thng do s bt cn ca
con ngi c th nh hng nghim trng ti sc khe ca c rn san h ni
chung. Tnh trng ca mt rn san h c lin quan rt cht ch vi cc h sinh
thi rng ngp mn v c bin xung quanh. Rng ngp mn v c bin lc cht

160

dinh dng t cc ngun trn t lin v l chic ni che ch v nui dng ca

nhiu sinh vt c tr rn san h.
Sarcophyton l mt chi san h mm sinh sng rt ph bin trn th gii ti
cc vng bin trn th gii ni chung v Vit Nam ni ring. Hin nay nc
ta hon ton cha c cng trnh khoa hc no c cng b v thnh phn ha
hc v hot tnh sinh hc ca cc loi thuc chi ny. Theo cc nghin cu v
thnh phn ha hc ca chi Sarcophyton, gn 200 hp cht c phn lp v
xc nh cu trc. Cc lp cht chnh ca chi ny l steroit v ditecpenoit (c
bit l cc cembranoit itecpen).
Di y, chng ti gii thiu mt s hp cht i din c phn lp
t chi Sarcophyton.
18

16
19
1

15

11

17

14

20

Sarcophytonin A [168]

Cembrene C [168]
18

18

16

19
1
11

16

19
1

15

14

11

17

20

20

Deoxosarcophine [168]
H

13-Dehydroxysarcoglaucol (2,5,6,9,10,13,14,16octahydro-4,12,15-trimethylcyclotetradeca[b]furan-8-arboxylic acid methyl ester) [169]

28

21
25

OH

17

21

26
24
20

18

18

OH

17

27

9
10

OH

25

20

13

11

19

10

HO

26
24

OH

27

13

COOCH3

Sarcophytonin B [168]

19

15

14

17

Sarcophine [169]

11

OH

HO

OH

OH

Cholestane-1,3,5,6-tetrol [170]

Ergostane-3,5,6,25,26-pentol [171]
26

28

21
24
18

20

21

25

19

13

11

27

OH

HO

20
17

19

10
3

26
24

18

17

13

11

25

27

9
10

HO

Papakusterol [171]

OH

OH

ergosta-1,3,5,6-tetraol [172]

161

III.1.2. Phn lp cc hp cht t san h mm Sarcophyton mililatensis

Mu san h mm ti (20 kg) c xay nh v chit 3 ln bng MeOH
nng (50oC mi ln 3h) v ct loi dung mi di p sut gim thu c 200 g
cn dch MeOH. Cn ny c ha vo nc ct v chit phn on ln lt vi
cc dung mi cloroform v n-butanol. Cn dch cloroform (150 g) c ha vo
metanol v chit vi hexan thu c cc cn dch: cn tan trong hexan (100 g) v
tan trong metanol (50 g). Cn tan trong hexan c tin hnh cha ct th trn
sc k ct pha thng ( = 10 cm, L = 50 cm) gradient nng axtn trong
hexan t 0 n 100% thu c 5 phn on nh hn k hiu l H1 - H5. Phn
on H2 (70g) c tin hnh trn sc k ct pha o vi h dung mi ra gii l
MeOH/nc 4/1 thu c cc hp cht SM6 (50 g) v SM7 (0,5 g) (S
III.1.2.a).
Phn on H4 (11 g) c tip tc phn ct trn sc k ct pha thng vi
h dung mi ra gii l hexan/axeton 7/1 thu c 5 phn on nh hn na k
hiu l H4A n H4E. T phn on nh H4B (3g) cc hp cht SM1 (40 mg),
SM4 (20 mg) v SM5 (9 mg) c lm sch bng phng php sc k ct silica
gel pha o vi h dung mi ra gii l MeOH/nc 5/1. Phn on H4D (1 g)
uc tin hnh phn lp trn sc k ct pha thng vi h dung mi ra gii
hexan/cloroform/MeOH 10/1/0.5 thu c cc hp cht SM2 (15 mg) v SM3
(20 mg) (S III.1.2.b).
San h mm Sarcophyton
mililatensis ti (20 kg)
- Chit MeOH, ct loi dung mi di p sut gim
- Loi mui bng cch chy qua ct Dianion vi
dung mi ra gii l nc ct, sau thu hi cht
bng MeOH.

B sung nc

SM - Cn chit MeOH

B sung cloroform
cloroform: nc 1/1

Lp nc
B sung BuOH

C - Cn cloroform (150 g)

BuOH: nc 1/1

B- Cn BuOH (20 g)

Lp nc

N - Cn nc (30 g)

B sung MeOH
B sung hexan

MeOH: hexan 1/1

M - Cn tan trong
MeOH (50 g)

H - Cn tan trong
hexan (100 g)

S III.1.2.a. S chit phn on mu san h mm

162

Cn H - (100 g)
Silica gel CC
Hexan/Axeton 100/0-0/100

H1 (5 g)

H2 (70 g)

H3 (9 g)

H4 (11 g)

H5 (5 g)

Silica gel CC pha o

MeOH/H2O 4/1

SM-6
(50 g)

H4A
(1 g)

Silica gel CC pha thng

n-hexan/axeton 7/1

SM-7
(0,5 g)

H4B
(3 g)

H4C
(2 g)

H4D
(1 g)

H4E
(4 g)

Silica gel CC pha o

MeOH/H2O 5/1

SM1
(40 mg)

SM4
(20 mg)

SM5
(9 mg)

SM2
(15 mg)

Silica gel CC pha thng

hexan/cloroform/MeOH
10/1/0.5

SM3
(20 mg)

S III.1.2.b. S phn lp cc hp cht t cn chit H

163

Cn chit M (50 g) c tin hnh chy ct th trn sc k ct pha thng

gradient nng metanol trong cloroform thu c 5 phn on nh hn l M1 M5. Phn on M2 (2g) c tin hnh phn tc trn sc k ct pha o vi h
dung mi ra gii MeOH/nc 5/1 thu c hp cht SM8 (20 mg). Bng phng
php tng t, cc hp cht SM9 (200 mg) v SM10 (100 mg) c phn lp t
phn on M3 (5 g) (S III.1.2.c).
Cn M - (50 g)

Silica gel CC
Cloroform/MeOH 100/0-0/100

M1 (20g)

M2 (2 g)

Silica gel CC pha

o MeOH/H2O: 5/1

M3 (5 g)

M4 (10 G)

M5 (13 g)

Silica gel CC pha

o MeOH/H2O: 5/1

SM8
(20 mg)
SM9
(200 mg)

SM10
(100 mg)

S III.1.2.c. S phn lp cc hp cht t cn chit M

III.1.3. Hng s vt l v d kin ph ca cc hp cht

III.1.3.1. Hp cht SM1: (-)-7-hydroxy-8-methoxydeepoxysarcophytoxide
(cht mi)
o
Cht du khng mu; []25
D -31 (CHCl3; c 0.50); positive ESI-MS m/z: 357
[M+Na]+, 317 [M-H2O+H]+, 303 [M-MeOH+H]+, 285 [M-MeOH-H2O+H]+
(C21H34O3).

H-NMR (500 MHz, CDCl3) (ppm): 5,36 (1H, d, J = 10,0 Hz, H-2),
5,22 (1H, d, J = 10,0 Hz, H-3), 2,22 (1H, m, Ha-5), 2,18 (1H, m, Hb-5), 1,77 (1H,
m, Ha-6), 1,55 (1H, m, Hb-6), 3,38 (1H, t, J = 8,0 Hz, H-7), 1,70 (1H, m, Ha-9),
1,65 (1H, m, Hb-9), 2,15 (1H, m, Ha-10), 1,99 (1H, m, Hb-10), 5,36 (1H, t, J =
7,0 Hz, H-11), 2,29 (1H, m, Ha-13), 2,08 (1H, m, Hb-13), 2,39 (1H, m, Ha-14),
1,64 (1H, m, Hb-14), 4,56 (1H, dd, J = 11,5, 4,0 Hz, Ha-16), 4,45 (1H, d, J = 11,5
Hz, Hb-16), 1,69 (3H, s, H-17), 1,81 (3H, s, H-18), 1,09 (3H, s, H-19), 1,68 (3H,
s, H-20) v 3,19 (3H, s, H-21).
1

164

C-NMR (125 MHz, CDCl3) (ppm): 33,97 (C-1), 85,80 (C-2), 124,47
(C-3), 142,60 (C-4), 38,66 (C-5), 29,01 (C-6), 74,03 (C-7), 79,57 (C-8), 31,97 (C9), 21,63 (C-10), 126,38 (C-11), 136,54 (C-12), 38,21 (C-13), 24,41 (C-14),
128,00 (C-15), 78,46 (C-16), 10,67 (C-17), 16,49 (C-18), 18,49 (C-19), 16,08 (C20) v 48,96 (C-21).
13

III.1.3.2. Hp cht SM2: (-)-17-Hydroxysarcophytonin A (cht mi)

o
Cht du khng mu; []25
D -97 (CDCl3; c 1.00); positive ESI-MS m/z: 325
[M+Na]+, 303 [M+H]+, 285 [M-H2O+H]+ (C20H30O2).

H-NMR (500 MHz, CDCl3) (ppm): 5,56 (1H, d, J = 10,0 Hz, H-2),
5,10 (1H, d, J = 10,0 Hz, H-3), 1,91 (1H, m, Ha-5), 2,12 (1H, m, Hb-5), 2,09 (1H,
m, Ha-6), 2,38 (1H, m, Hb-6), 4,85 (1H, t, J = 7,0 Hz, H-7), 2,18 (1H, m, Ha-9),
2,31 (1H, m, Hb-9), 2,06 (2H, m, H-10), 5,01 (1H, t, J = 7,0 Hz, H-11), 1,95 (2H,
m, H-13), 1,93 (1H, m, Ha-14), 2,45 (1H, m, Hb-14), 4,66 (1H, dd, J = 11,5, 3,0
Hz, Ha-16), 4,75 (1H, dd, J = 11,5, 4,0 Hz, Hb-16), 4,24 (1H, d, J = 11,0 Hz, Ha17), 4,28 (1H, d, J = 11,0 Hz, Hb-17), 1,70 (3H, s, H-18), 1,58 (3H, s, H-19) v
1,59 (3H, s, H-20).
1

C-NMR (125 MHz, CDCl3) (ppm): 138,23 (C-1), 84,28 (C-2), 124,85
(C-3), 141,02 (C-4), 39,14 (C-5), 24,71 (C-6), 125,52 (C-7), 133,34 (C-8), 40,22
(C-9), 23,42 (C-10), 124,21 (C-11), 134,92 (C-12), 37,38 (C-13), 25,87 (C-14),
130,92 (C-15), 75,74 (C-16), 56,84 (C-17), 15,05 (C-18), 15,07 (C-19) v 15,63
(C-20).
13

III.1.3.3. Hp cht SM3: Sarcophytol V (cht mi)

o
Cht du khng mu; []25
D -45 (CDCl3; c 1.00); positive ESI-MS m/z: 327
[M+Na]+, 287 [M-H2O+H]+, 269 [M-2H2O+H]+ (C20H32O2).

H-NMR (500 MHz, CDCl3) (ppm): 6,39 (1H, d, J = 11,0 Hz, H-2), 5,91
(1H, d, J = 11,0 Hz, H-3), 2,18 (2H, m, H-5), 2,22 (2H, m, H-6), 4,99 (1H, t, J =
7,0 Hz, H-7), 2,10 (2H, m, H-9), 2,15 (2H, m, H-10), 4,99 (1H, t, J = 7,0 Hz, H11), 2,14 (2H, m, H-13), 2,30 (1H, t, J = 7,0 Hz, H-14), 3,43 (1H, d, J = 11,0 Hz,
Ha-16), 3,63 (1H, d, J = 11,0 Hz, Hb-16), 1,31 (3H, s, H-17), 1,77 (3H, s, H-18),
1,54 (3H, s, H-19) v 1,62 (3H, s, H-20).
1

C-NMR (125 MHz, CDCl3) (ppm): 142,97 (C-1), 121,47 (C-2), 120,47
(C-3), 138,63 (C-4), 38,79 (C-5), 24,85 (C-6), 125,55 (C-7), 134,13 (C-8), 38,97
(C-9), 24,38 (C-10), 125,50 (C-11), 134,64 (C-12), 41,18 (C-13), 26,42 (C-14),
76,39 (C-15), 69,29 (C-16), 24,62 (C-17), 17,84 (C-18), 15,67 (C-19) v 16,00
(C-20).
13

III.1.3.4. Hp cht SM4: Sarcophytol X (cht mi)

o
Cht du khng mu; []25
D -90 (CDCl3; c 1.00); positive ESI-MS m/z: m/z 334.8
[M-H2O+H]+, 316.8 [M-2H2O+H]+, 298.8 [M-3H2O+H]+ (C20H32O5).

H-NMR (500 MHz, CDCl3) (ppm): 6,49 (1H, d, J = 1,0 Hz, H-3), 2,35
(2H, m, H-5), 1,70 (1H, m, Ha-6), 2,02 (1H, m, Hb-6), 2,87 (1H, dd, J = 8,0, 3,5
1

165

Hz, H-7), 1,46 (1H, m, Ha-9), 1,97 (1H, m, Hb-9), 2,08 (2H, m, H-10), 4,98 (1H,
dt, J = 7,0, 1,0 Hz, H-11), 2,15 (1H, m, Ha-13), 2,22 (1H, m, Hb-13), 1,65 (1H,
m, Ha-14), 2,48 (1H, m, Hb-14), 3,43 (1H, Ha-16), 3,48 (1H, Hb-16), 1,44 (3H, s,
H-17), 2,11 (3H, d, J = 1,0 Hz, H-18), 1,24 (3H, s, H-19) v 1,55 (3H, d, J = 1,0
Hz, H-20).
C-NMR (125 MHz, CDCl3) (ppm): 71,95 (C-1), 199,42 (C-2), 122,86
(C-3), 157,89 (C-4), 38,44 (C-5), 25,59 (C-6), 61,77 (C-7), 60,87 (C-8), 37,11 (C9), 22,82 (C-10), 126,57 (C-11), 134,59 (C-12), 34,91 (C-13), 27,41 (C-14),
65,79 (C-15), 65,92 (C-16), 15,74 (C-17), 19,13 (C-18), 17,98 (C-19) v 15,25
(C-20).
13

III.1.3.5. Hp cht SM5: (+)-7,8-dihydroxydeepoxysarcophytoxide

mi)

(cht

Cht du khng mu; []25

D +105 (CDCl3; c 1.00); positive ESI-MS: m/z 343
[M+Na]+, 303 [M-H2O+H]+ (C20H32O3).
H-NMR (500 MHz, CDCl3) (ppm): 5,40 (1H, d, J = 11,0 Hz, H-2), 5,24
(1H, d, J = 11,0 Hz, H-3), 2,24 (2H, m, H-5), 1,80 (1H, m, Ha-6), 1,61 (1H, m,
Hb-6), 3,43 (1H, t, J = 8,0 Hz, H-7), 1,77 (1H, m, Ha-9), 1,67 (1H, m, Hb-9), 2,21
(1H, m, Ha-10), 2,11 (1H, m, Hb-10), 5,20 (1H, t, J = 7,0 Hz, H-11), 2,17 (1H, m,
Ha-13), 2,05 (1H, m, Hb-13), 2,20 (1H, m, Ha-14), 1,93 (1H, m, Hb-14), 4,44 (1H,
d, J = 11,0 Hz, Ha-16), 4,53 (1H, dd, J = 11,0, 4,0 Hz, Hb-16), 1,67 (3H, s, H17), 1,84 (3H, s, H-18), 1,20 (3H, s, H-19) v 1,64 (3H, s, H-20).
1

C-NMR (125 MHz, CDCl3) (ppm): 133,90 (C-1), 85,14 (C-2), 125,16
(C-3), 141,72 (C-4), 36,30 (C-5), 29,82 (C-6), 74,39 (C-7), 75,44 (C-8), 38,32 (C9), 22,78 (C-10), 125,20 (C-11), 136,59 (C-12), 37,50 (C-13), 23,83 (C-14),
128,20 (C-15), 78,41 (C-16), 10,51 (C-17), 17,63 (C-18), 24,65 (C-19) v 16,04
(C-20).
13

III.1.3.6. Hp cht SM6: (+)-Sarcophine

o
Cht du khng mu; []25
D +95 (CHCl3; c 1.00); positive ESI-MS m/z: 339
[M+Na]+, 317 [M+H]+, 299 [M-H2O+H]+ (C20H28O3).

H-NMR (500 MHz, CDCl3) (ppm): 5,45 (1H, d, J = 9,0 Hz, H-2), 4,99
(1H, d, J = 9,0 Hz, H-3), 2,31 (1H, m, Ha-5), 2,22 (1H, m, Hb-5), 1,83 (1H, m,
Ha-6), 1,72 (1H, m, Hb-6), 2,66 (1H, t, J = 5,5 Hz, H-7), 2,05 (1H, m, Ha-9), 1,19
(1H, dt, J = 13,0 and 4,0, Hz, Hb-9), 2,21 (1H, m, Ha-10), 1,94 (1H, m, Hb-10),
5,11 (1H, t, J = 7,0 Hz, H-11), 2,34 (1H, m, Ha-13), 1,92 (1H, m, Hb-13), 2,36
(1H, m, Ha-14), 1,71 (1H, m, Hb-14), 1,86 (3H, s, H-17), 1,93 (3H, s, H-18), 1,27
(3H, s, H-19) v 1,61 (3H, s, H-20).
1

C-NMR (125 MHz, CDCl3) (ppm): 162,8 (C-1), 79,8 (C-2), 119,2 (C3), 144,9 (C-4), 38,4 (C-5), 25,7 (C-6), 61,8 (C-7), 60,1 (C-8), 36,9 (C-9), 23,0
(C-10), 124,7 (C-11), 134,5 (C-12), 35,1 (C-13), 26,8 (C-14), 123,2 (C-15), 174,9
(C-16), 9,2 (C-17), 16,7 (C-18), 18,1 (C-19) v 15,5 (C-20).
13

166

III.1.3.7. Hp cht SM7: Sarcophytoxide

o
Cht du khng mu; []25
D -65 (CHCl3; c 1,00); positive ESI-MS m/z: 325
[M+Na]+, 303 [M+H]+, 285 [M-H2O+H]+ (C20H30O2).

H-NMR (500 MHz, CDCl3) (ppm): 5,41 (1H, m, H-2), 5,27 (1H, d, J =
10,0 Hz, H-3), 2,33 (1H, m, Ha-5), 2,24 (1H, m, Hb-5), 1,81 (1H, m, Ha-6), 1,71
(1H, m, Hb-6), 2,70 (1H, t, J = 5,5 Hz, H-7), 2,07 (1H, m, Ha-9), 1,15 (1H, dt, J
= 13,0 and 4,0 Hz, Hb-9), 2,21 (1H, m, Ha-10), 1,96 (1H, m, Hb-10), 5,13 (1H, t,
J = 7,5 Hz, H-11), 2,25 (1H, m, Ha-13), 1,93 (1H, m, Hb-13), 2,35 (1H, m, Ha14), 1,66 (1H, m, Hb-14), 4,46 (1H, dd, J = 11,5 and 3,5 Hz), Ha-16), 4,53 (1H,
dd, J = 11,5 and 4,5 Hz), Hb-16), 1,66 (3H, s, H-17), 1,86 (3H, s, H-18), 1,27
(3H, s, H-19) v 1,58 (3H, s, H-20).
1

C-NMR (125 MHz, CDCl3) (ppm): 133,9 (C-1), 85,3 (C-2), 125,4 (C3), 140,4 (C-4), 35,0 (C-5), 27,4 (C-6), 62,4 (C-7), 60,2 (C-8), 38,9 (C-9), 23,2
(C-10), 123,4 (C-11), 135,9 (C-12), 38,6 (C-13), 23,7 (C-14), 128,1 (C-15), 78,3
(C-16), 10,4 (C-17), 17,7 (C-18), 16,4 (C-19) v 15,4 (C-20).
13

III.1.3.8. Hp cht SM8: Sarcomilasterol (cht mi)

o
Tinh th hnh kim mu trng; im chy 160-161 oC; []20
D +11 (CHCl3; c
+
+
0,24); positive ESI-MS m/z: m/z 471.1 [M+Na] , 449.1 [M+H] , 431.1 [MH2O+H]+ (C28H48O4).

H-NMR (500 MHz, CDCl3+CD3OD) (ppm): 1,84 (1H, m, Ha-1), 1,42

(1H, m, Hb-1), 1,87 (1H, m, Ha-2), 1,42 (1H, m, Hb-2), 3,53 (1H, m, H-3), 2,24
(1H, m, Ha-4), 1,44 (1H, m, Hb-4), 1,75 (1H, m, H-5), 4,21 (1H, d, J = 10,0 Hz,
H-6), 6,61 (1H, d, J = 4,0 Hz, H-7), 3,82 (1H, m, Ha-11), 3,63 (1H, m, Hb-11),
1,60 (1H, m, Ha-12), 1,08 (1H, m, Hb-12), 3,34 (1H, t, J = 10,0 Hz, H-14), 1,58
(1H, m, Ha-15), 1,40 (1H, m, Hb-15), 1,82 (1H, m, Ha-16), 1,58 (1H, m, Hb-16),
1,67 (1H, m, H-17), 0,65 (3H, s, H-18), 1,12 (3H, s, H-19), 1,36 (1H, m, H-20),
0,97 (3H, d, J = 7,0 Hz, H-21), 1,40 (1H, m, Ha-22), 0,93 (1H, m, Hb-22), 1,42
(1H, m, Ha-23), 0,93 (1H, m, Hb-23), 1,18 (1H, m, H-24), 1,55 (1H, m, H-25),
0,79 (3H, d, J = 7,0 Hz, H-26), 0,86 (3H, d, J = 7,0 Hz, H-27) v 0,78 (3H, d, J
= 7,0 Hz, H-28).
1

C-NMR (125 MHz, CDCl3+CD3OD) (ppm): 31,95 (C-1), 30,21 (C-2),

69,72 (C-3), 32,48 (C-4), 48,54 (C-5), 68,80 (C-6), 148,57 (C-7), 136,63 (C-8),
206,08 (C-9), 44,99 (C-10), 59,06 (C-11), 40,89 (C-12), 46,14 (C-13), 42,75 (C14), 26,60 (C-15), 26,27 (C-16), 49,70 (C-17), 17,42 (C-18), 16,13 (C-19), 35,34
(C-20), 18,96 (C-21), 33,14 (C-22), 31,40 (C-23), 39,14 (C-24), 31,58 (C-25),
17,67 (C-26), 20,51 (C-27) v 15,49 (C-28).
13

III.1.3.9. Hp cht SM9: Sarcoaldesterol B

o
Cht bt mu trng; []25
D : -26 (c 1,5, MeOH); positive ESI-MS m/z:
473,1 [M+Na]+, 433,1 [M-H2O+H]+, 415,1 [M-2H2O+H]+, 397,1 [M-3H2O+H]+
(C28H50O4).

167

H-NMR (500 MHz, CD3OD) (ppm): 2,08 (1H, m, Ha-1), 1,81 (1H, m,
Hb-1), 1,77 (1H, m, Ha-2), 1,52 (1H, m, Hb-2), 3,84 (1H, m, H-3), 2,09 (1H, m,
Ha-4), 1,57 (1H, m, Hb-4), 3,46 (1H, br s, H-6), 1,81 (1H, m, Ha-7), 1,52 (1H, m,
Hb-7), 1,80 (1H, m, H-8), 1,47 (1H, m, H-9), 4,01 (1H, m, H-11), 2,28 (1H, m,
Ha-12), 1,26 (1H, m, Hb-12), 1,21 (1H, m, H-14), 1,62 (1H, m, Ha-15), 1,15 (1H,
m, Hb-15), 1,90 (1H, m, Ha-16), 1,32 (1H, m, Hb-16), 1,21 (1H, m, H-17), 0,75
(3H, s, H-18), 1,29 (3H, s, H-19), 1,40 (1H, m, H-20), 0,99 (3H, d, J = 7,0 Hz,
H-21), 1,47 (1H, m, Ha-22), 1,01 (1H, m, Hb-22), 1,45 (1H, m, Ha-23), 1,01 (1H,
m, Hb-23), 1,23 (1H, m, H-24), 1,60 (1H, m, H-25), 0,83 (3H, d, J = 7,0 Hz, H26), 0,90 (3H, d, J = 7,0 Hz, H-27) v 0,82 (3H, d, J = 7,0 Hz, H-28).
1

C-NMR (125 MHz, CD3OD) (ppm): 35,3 (C-1), 32,0 (C-2), 69,4 (C-3),
41,9 (C-4), 77,4 (C-5), 76,5 (C-6), 35,1 (C-7), 30,3 (C-8), 53,0 (C-9), 40,9 (C-10),
68,2 (C-11), 52,7 (C-12), 44,2 (C-13), 56,5 (C-14), 25,3 (C-15), 29,3 (C-16), 57,4
(C-17), 13,4 (C-18), 17,5 (C-19), 37,4 (C-20), 19,4 (C-21), 34,8 (C-22), 31,7 (C23), 40,3 (C-24), 32,7 (C-25), 18,1 (C-26), 20,9 (C-27) v 16,0 (C-28).
13

III.1.3.10. Hp cht SM10: Ergosta-1,3,5,6-tetraol

o
Tinh th hnh kim mu trng; []25
D -13 (CHCl3; c 1,20); positive ESI-MS
+
+
m/z: 473,1 [M + Na] , 450,9 [M +H] (C28H50O4).

H-NMR (500 MHz, CD3OD) (ppm): 3,97 (1H, dd, J = 10,5, 4,5 Hz, H1), 2,01 (1H, m, Ha-2), 1,57 (1H, m, Hb-2), 4,03 (1H, m, H-3), 2,07 (1H, dd, J =
12,5, 11,5 Hz, H-3), 3,45 (1H, br s, H-6), 1,48 (1H, m, Ha-7), 1,00 (1H, m, Hb-7),
1,77 (1H, m, H-8), 1,65 (1H, m, H-9), 2,15 (1H, m, Ha-11), 1,42 (1H, m, Hb-11),
2,01 (1H, m, Ha-12), 1,21 (1H, m, Hb-12), 1,15 (1H, m, H-14), 1,61 (1H, m, Ha15), 1,17 (1H, m, Hb-15), 1,86 (1H, m, Ha-16), 1,29 (1H, m, Hb-16), 1,15 (1H, m,
H-17), 0,73 (3H, s, H-18), 1,15 (3H, s, H-19), 1,40 (1H, m, H-20), 0,95 (3H, d, J
= 7,0 Hz, H-21), 1,73 (1H, m, Ha-22), 1,54 (1H, m, Hb-22), 1,45 (1H, m, Ha-23),
1,00 (1H, m, Hb-23), 1,23 (1H, m, H-24), 1,61 (1H, m, H-25), 0,84 (3H, d, J =
7,0 Hz, H-26), 0,90 (3H, d, J = 7,0 Hz, H-27) v 0,83 (3H, d, J = 7,0 Hz, H-28).
1

C-NMR (125 MHz, CD3OD) (ppm): 74,3 (C-1), 42,5 (C-2), 66,0 (C-3),
41,5 (C-4), 77,5 (C-5), 77,1 (C-6), 34,9 (C-7), 32,1 (C-8), 47,4 (C-9), 44,9 (C-10),
25,0 (C-11), 42,0 (C-12), 43,3 (C-13), 57,5 (C-14), 25,6 (C-15), 29,1 (C-16), 57,7
(C-17), 12,6 (C-18), 10,2 (C-19), 37,6 (C-20), 19,4 (C-21), 35,2 (C-22), 31,7 (C23), 40,4 (C-24), 32,7 (C-25), 18,1 (C-26), 20,9 (C-27) v 15,9 (C-28).
13

168

III.1.4. Kt qu xc nh cu trc ca cc hp cht

III.1.4.1. Hp cht SM1: (-)-7-hydroxy-8-methoxydeepoxysarcophytoxide
(cht mi)
Hp cht SM1 c phn lp di dng cht du khng mu. Cng thc
phn t ca n c xc nh l C21H34O3 bng cc d kin ph khi lng
(positive ESI-MS m/z: 357 [M+Na]+, 317 [M-H2O+H]+, 303 [M-MeOH+H]+, 285
[M-MeOH-H2O+H]+) v ph cng hng t ht nhn (Bng III.1.4.1).

Hnh III.1.4.1.a. Ph 1H-NMR ca SM1

18

4
7

HO

16

21

OMe

19

15

17

14

11
20

Hnh III.1.4.1.b. Cu trc ha hc ca SM1

Ph 1H-NMR ca SM1 xut hin cc tn hiu c trng ca hai proton
olefin cng hng ti 5,22 (1H, d, J = 11,0 Hz, H-3) v 5,36 (1H, d, J = 11,0
Hz, H-11); hai proton oximetin ti 5,38 (1H, d, J = 11,0 Hz, H-2) v 3,38 (1H,
t, J = 7,0 Hz, H-7); cc proton ca nhm oximetilen ti 4,45 (1H, d, J = 11,0
Hz, Ha-16)/4,56 (1H, dd, J = 11,0 v 4,0 Hz, Hb-16); cc proton ca 4 nhm
metyl bc bn ti 1,69 (3H, s, H-17), 1,81 (3H, s, H-18), 1,09 (3H, s, H-19) v
1,67 (3H, s, H-20); mt nhm metoxy ti 3,19 (3H, s, H-21) v cc tn hiu
proton ca cc 6 nhm metilen (Bng III.1.4.1).

169

Hnh III.1.4.1.c. Ph 13C-NMR v cc ph DEPT ca SM1

Bng III.1.4.1. Kt qu ph NMR ca SM1

C

C *[173]

Ca, b

1
2
3
4
5
6
7

38,8
28,3
73,7

133,97
85,80
124,47
142,60
38,66
29,01
74,03

Ha, c
multiplicity (J in Hz)
5,36 d (11,0)
5,22 d (11,0)
2,22 m/2,18 m
1,77 m/1,55 m
3,38 t (7,0)
170

HMBC
(HC)

5, 18

5, 6, 8, 19

8
9
10
11
12
13
14
15
16

79,3
31,5
21,3
127,9
135,7
36,8
26,9
-

79,57
31,97
21,63
126,38
136,54
38,21
24,41
128,00
78,46

17
18
19
20
21

9,6
16,7
18,1
16,2
48,9

10,67
16,49
18,49
16,08
48,96

1,70 m/1,65 m
2,15 m/1,99 m
5,36 t (7,0)
2,29 m/2,08 m
2,39 m/1,64 m
4,56 dd (11,0, 4,0)
4,45 d (11,0)
1,69 s
1,81 s
1,09 s
1,68 s
3,19 s

10
10, 13, 20
1, 11, 12, 14
12

1, 15, 16
3, 4, 5
7, 8, 9
11, 12, 13
8

o trong CDCl3, b125 MHz, c500 MHz, *s liu ph ca hp cht (-)-7-Hydroxy-8-methoxydeepoxysarcophine

Hnh III.1.4.1.d. Ph HSQC ca SM1

Trn ph 13C-NMR ca SM1 xut hin 20 tn hiu c trng ca mt hp
cht ditecpen dng cembranoit. Trong , cc tn hiu cng hng ti 124,47
(CH, C-3)/142,60 (C, C-4), 126,38 (CH, C-11)/136,54 (C, C-12) v 133,97 (C, C1)/128,0 (C, C-15) khng nh s tn ti ca hai lin kt i b th 3 v tr v mt
lin kt i b th hon ton. Bn nhm metyl bc ba v mt nhm metoxy c
khng nh tng ng bi cc tn hiu cng hng ti 10,67 (C-17), 16,49 (C18), 18,49 (C-19), 16,08 (C-20) v 48,96 (C-21). Ngoi ra cc tn hiu c trng
cho hai nhm oximetin, mt nhm oximetilen v mt nhm ete bc bn cng

171

c khng nh tng ng ti 85,80 (C-2), 74,03 (C-7), 78,46 (C-16) v 79,57

(C-8). Cc tn hiu cacbon c gn vi cc tn hiu proton tng ng thng qua
ph cng hng t ht nhn hai chiu tng tc trc tip HSQC (Bng III.1.4.1 v
Hnh III.1.4.1.d).

Hnh III.1.4.1.e. Ph HMBC ca SM1

18

HO

H
O

16

21

OMe

19

15

11
17

14

HMBC
20

COSY

Hnh III.1.4.1.f. Cc tng tc HMBC v COSY chnh ca SM1

Trn ph 1H-1H COSY xut hin cc tng tc gia H-2/H-3, H2-5/H26/H-7, H2-9/H2-10/H-11 v H2-13/H2-14 (Hnh III.1.4.1.f). Kt qu trn, cng vi
cc tn hiu tng tc HMBC gia H-2/C-1, H3-18/C-3, H3-18/C-4, H3-18/C-5,
H3-19/C-7, H3-19/C-8, H3-19/C-9, H3-20/C-11, H3-20/C-12, H3-20/C-13 v H213/C-1 cho php ghp ni cc phn cu trc t C-1 n C-14 ca vng 14 cacbon.

172

Hnh III.1.4.1.g. Ph 1H-1H COSY ca SM1

Hnh III.1.4.1.h. Ph ROESY ca SM1

V tr ca nhm metyl C-17 c khng nh bi cc tn hiu tng tc t
cc proton ca n ti C-1, C-15 v C-16 nhn c trn ph HMBC. Cng vi cc

173

phn tch nu phn trn cho php khng nh ton b cu trc phng ca
SM1 nh a ra Hnh III.1.4.1.b.
CH3

H
O

HO

H
H

CH3

H 3C
CH3
OCH3

Hnh III.1.4.1.i. Cu hnh khng gian v cc tng tc

ROESY then cht ca SM1
Cu hnh ti cc v tr bt i ca SM1 c xc nh bng cch so snh
s liu ph NMR ca n vi cc hp cht c cu trc tng t c cng b
[168, 173]v c khng nh thm bng cc tng tc trn ph ROESY. Cu
hnh ca cc lin kt i C-3/C-4 v C-11/C-12 c khng nh u dng trans
da trn s liu chuyn dch ha hc 13C ca cc cacbon C-18 v C-20 tng
ng ti 16,49 v 16,08 (nh hn 20 ppm) [174]

Hnh III.1.4.1.j. Ph ESI-MS ca SM1

Trn ph ROESY ca SM1, tn hiu tng tc cho gia proton H-2 vi
cc proton H3-18 cho php khng nh chng u nm v tr v cu hnh ca
lin kt i C-3/C-4 dng trans. Cc tn hiu tng tc cho ROESY gia H-3
v H-7, H-3 v Ha-6 ( 1.77), Ha-6 v H3-21 cho thy H-3, Ha-6 v H3-21 u
nm v tr . T tt c cc phn tch nu trn, cu trc ha hc ca SM1 c
xc nh cho mt hp cht mi l (-)-7-hydroxy-8-methoxydeepoxysarcophytoxide.
174

III.1.4.2. Hp cht SM2: (-)-17-Hydroxysarcophytonin A

Hp cht SM2 cng c phn lp di dng cht du khng mu. T cc
d kin ph khi lng (positive ESI-MS m/z: 324,9 [M+Na]+, 302,9 [M+H]+,
284,8 [M-H2O+H]+) v ph cng hng t ht nhn (xem Bng III.1.4.2) cho
php xc nh cng thc phn t ca n l C20H30O2.

Hnh III.1.4.2.a. Ph 1H-NMR ca SM2

18

H
O

16

19
1

15

11
14

17

OH

20

Hnh III.1.4.2.b. Cu trc ha hc ca SM2

Ph 1H- v 13C-NMR ca SM2 c dng c trng cho mt hp cht
ditecpen dng cembranoit. Trn ph 1H-NMR xut hin cc tn hiu c trng
ca: ba proton olefin ti 5,10 (1H, d, J = 10,0 Hz, H-3), 4,85 (1H, t, J = 7,0 Hz,
H-7) v 5,01 (1H, t, J = 7,0 Hz, H-11); mt proton oximetin ti 5,56 (1H, d, J =
10.0 Hz, H-2); v cc proton oximetilen ti 4,66 (1H, dd, J = 11,5 v 3,0 Hz,
Ha-16), 4,75 (1H, dd, J = 11,5 v 4,0 Hz, Hb-16), 4,24 (1H, d, J = 11,0 Hz, Ha17), 4,28 (1H, d, J = 11,0 Hz, Hb-17). Ngoi ra, s xut hin ca bn nhm metyl
gn vo ni i cng c khng nh bi s xut hin cc tn hiu cng hng
ti 1,70 (3H, s, H-18), 1,58 (3H, s, H-19) v 1,59 (3H, s, H-29).

175

Hnh III.1.4.2.c. Ph 13C-NMR ca SM2

Hnh III.1.4.2.d. Ph 13C-NMR v cc ph DEPT ca SM2

Ph 13C-NMR ca SM2 xut hin 20 tn hiu c trng ca mt hp cht
ditecpen dng cembranoit. Trong , s xut hin ca ba lin kt i b th 3 v
tr c khng nh bi cc tn hiu cng hng ti 124,85 (CH, C-3)/141,02
(C, C-4), 125,52 (CH, C-7)/133,34 (C, C-8) v 124,21 (CH, C-11)/134,92 (C, C12); mt lin kt i b th hon ton ti 138,23 (C, C-1)/130,92 (C, C-15); s
176

xut hin ca mt nhm oximetin v hai nhm oximetilen c c trng tng

ng bi cc tn hiu cng hng ti 84,28 (CH, C-2), 75,74 (CH2, C-16) v
56,84 (CH2, C-17). Cc tn hiu metyl cng hng ti 15,05 (C-18), 15,07 (C19) v 15,63 (C-20) c trng cho cc nhm metyl gn vo cc lin kt i. Da
trn chuyn dch ha hc ca cc nhm metyl ny u nh hn 20 ppm cho
php khng nh cc lin kt i m chng gn vo u nm dng trans [174].

Hnh III.1.4.2.e. Ph HSQC ca SM2

Tt c cc tn hiu carbon c gn vi tn hiu proton tng ng thng
qua ph cng hng t ht nhn hai chiu tng tc trc tip HSQC (Hnh
III.1.4.2.e) v kt qu chi tit c a ra trong Bng III.1.4.2.
Bng III.1.4.2. Kt qu ph NMR ca SM2
C

C* [175]

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13

135,4
84,0
125,8
140,3
39,2
24,8
125,5
133,2
40,4
23,5
124,0
134,0
37,2

138,23
84,28
124,85
141,02
39,14
24,71
125,52
133,34
40,22
23,42
124,21
134,92
37,38

Ha, c
multiplicity (J in Hz)
5,56 d (10,0)
5,10 d (10,0)
1,91 m/2,12 m
2,09 m/2,38 m
4,85 t (7,0)
2,18 m/2,31 m
2,06 m
5,01 t (7,0)
1,95 m
177

HMBC
(HC)

5, 18

13, 20

14
15
16

25,9
127,1
78,4

25,87
130,92
75,74

1,93 m/2,45 m
4,66 dd (11,5, 3,0)
4,75 dd (11,5, 4,0)

17

10,2

56,84

4,24 d (11,0)
4,28 d (11,0)

1, 2, 13
1,15
1, 15, 16

18
15,0
15,05
1,70 s
3, 4, 5
19
15,2
15,07
1,58 s
7, 8, 9
20
15,6
15,63
1,59 s
11, 12, 13
a
b
c
*
o trong CDCl3, 125 MHz, 500 MHz, s liu ca sarcophytonin A.

Cc s liu ph 1H v 13C-NMR ca SM2 hon ton tng t nh s liu

c cng b ca hp cht sarcophytonin A [175]. S khc bit d nhn thy
nht l s mt i mt nhm metyl v thay vo l s xut hin ca mt nhm
oximetilen trong phn t ca SM2 so vi sarcophytonin A. S liu ph NMR ti
tt c cc v tr ca SM2 c gn bng cch so snh vi cc s liu ca
sarcophitonin A v cc hp cht c cu trc tng t [175-176] v c khng
nh li bi cc tng tc xa trn ph HMBC (Bng III.1.4.2). Nhm oximetilen
xut hin thm c xc nh ti v tr C-17 bi cc tng tc xa nhn c trn
ph HMBC t cc proton ca n ti cc cacbon C-1, C-15 v C-16.
18

H
4

7
19

16

1
15

11
14

17

OH

20

Hnh III.1.4.2.f. Cc tng tc HMBC (HC) chnh ca SM2

Hnh III.1.4.2.g. Ph HMBC ca SM2

178

Hnh III.1.4.2.h. Ph ESI-MS ca SM2

T tt c cc phn tch nu trn, cu trc ha hc ca SM2 c xc
nh l (-)-17-Hydroxysarcophytonin A. y l mt hp cht c cu trc mi, ln
u tin c phn lp t thin nhin.
III.1.4.3. Hp cht SM3: sarcophytol V
Hp cht SM3 c phn lp i dng du khng mu. Cng thc phn
t ca n c xc nh l C20H32O2 bng cc kt qu ph khi lng (positive
ESI-MS: m/z 326.9 [M+Na]+, 286.9 [M-H2O+H]+, 268.9 [M-H2O+H]+) v ph
cng hng t ht nhn.

Hnh III.1.4.3.a. Ph 1H-NMR ca SM3

179

18

4
16

7
19
1
11
14

15

OH
OH
17

20

Hnh III.1.4.3.b. Cu trc ha hc ca SM3

Ph 1H-NMR ca SM3 xut hin cc tn hiu c trng ca 4 proton olefin
ti 6,39 (1H, d, J = 11,0 Hz, H-2), 5,91 (1H, d, J = 11,0 Hz, H-3) v 4,99 (2H,
t, J = 7,0 Hz, H-7 v H-11); cc proton ca mt nhm oximetilen ti 3,43 (1H,
d, J = 11,0 Hz, Ha-16) v 3,63 (1H, d, J = 11,0 Hz, Hb-16); v bn nhm metyl
ti 1,31 (3H, s, H-17), 1,77 (3H, s, H-18), 1,54 (3H, s, H-19) v 1,62 (3H, s, H20).

Hnh III.1.4.3.c. Ph 13C-NMR ca SM3

Ph 13C-NMR ca SM3 cng xut hin 20 tn hiu cacbon c trng ca
mt hp cht ditecpen dng cembranoit. Trong cc tn hiu c trng cho bn
lin kt i u b th ba v tr cng hng ti 142,97 (C-1)/121,47 (C-2),
120,47(C-3)/138,63 (C-4), 125,55 (C-7)/134,13 (C-8) v 125,50 (C-11)/134,64
(C-12); cc tn hiu tng ng ca mt cacbon bc 4 ni vi nguyn t xy v
mt nhm oximetilen cng hng ti 76,39 (C-15) v 69,29 (C-16). Bn nhm
metyl xut hin ti 24,62 (C-17), 17,84 (C-18), 15,67 (C-19) v 16,00 (C-20).
Thng qua ph HSQC, tt c cc tn hiu cacbon c gn vi cc proton tng
ng.

180

Hnh III.1.4.3.d. Ph 13C-NMR v cc ph DEPT ca SM3

Bng III.1.4.3.Kt qu ph NMR ca SM3
HMBC
Ha, c
mult, (J in Hz)
(HC)
1
144,4
147,1
142,97
2
121,4
118,7
121,47
6,39 d (11,0)
1, 3, 4, 14, 15
3
120,5
120,5
120,47
5,91 d (11,0)
1, 2, 5, 18
4
138,3
137,8
138,63
5
38,6
38,79
2,18 m
3, 4
6
24,7
24,85
2,22 m
4, 7, 8
7
125,6
125,55
4,99 t (7,0)
5, 6, 8, 9, 19
8
133,9
134,13
9
38,9
38,97
2,10 m
10
23,6
24,3
24,38
2,15 m
8, 9, 11, 12
11
125,7
125,3
125,50
4,99 t (7,0)
9, 10, 12, 13, 20
12
136,0
134,9
134,64
13
41,6
41,8
41,18
2,14 m
1, 11, 12, 14, 20
14
26,7
26,3
26,42
2,30 t (7,0)
1, 2, 12, 13, 15
15
76,6
76,39
16
29,7
3,43 d (11,0)
1, 15, 17
69,3
69,29
3,63 d (11,0)
17
29,7
1,31 s
1, 15, 16
24,7
24,62
18
18,2
17,8
17,84
1,77 s
3, 4, 5
19
15,6
15,67
1,54 s
7, 8, 9
20
16,5
15,7
16,00
1,62 s
11, 12, 13
a
b
c
*
o trong CDCl3, 125 MHz, 500 MHz, s liu ph ca 7,8-epoxy-1,3,11-cembratriene15R(),16-diol, #s liu ph ca 15-hydroxycembra-1,3,7,11-tetraene
C

C*[177]

C#[178]

Ca, b

181

Hnh III.1.4.3.e. Ph HSQC ca SM3

Hnh III.1.4.3.f. Ph HMBC ca SM3

182

18

OH
OH

16

7
19

15
1
14

11

17

20

Hnh III.1.4.3.g. Cc tng tc HMBC (HC)chnh ca SM3

Cc s liu ph NMR ca SM3 hon ton tng t nh ca hp cht 15hydroxycembra-1,3,7,11-tetraene (Bng III.1.4.3), ngoi tr s xut hin tn hiu
ca mt nhm oximetilen trong phn t ca SM3 thay v mt nhm metyl trong
phn t ca hp cht ny. iu cho php khng nh SM3 l mt dn xut
hydroxyl ha ca 15-hydroxycembra-1,3,7,11-tetraene. So snh trc tip s liu
ph 13C-NMR ca SM3 vi cc s liu ca hp cht ny [178] cho php d on
nhm hydroxyl c gn vo v tr C-16. iu ny c minh chng bi s ph
hp hon ton v s liu ph NMR ca SM3 vi cc hp cht c cng cu trc
cc v tr th ca cc nhm hydroxyl [177] v bng cc tng tc xa trn ph
HMBC gia cc proton H2-16 vi cc cacbon C-1, C-15 v C-17, v t proton H317 vi cc cacbon C-1, C-15 v C-16. S liu ph NMR ti tt c cc v tr trong
phn t ca SM3 c gn bng cch so snh trc tip vi cc s liu c
cng b ca hp cht 15-hydroxycembra-1,3,7,11-tetraene [178] v c khng
nh bi cc tng tc trn ph HMBC (Bng III.1.4.3). Cc lin kt i u c
khng nh dng trans da vo gi tr chuyn dch ha hc 13C-NMR ca
cc nhm metyl nh vo chng u nh hn 20 ppm [174]. Cu hnh ca nhm
15-hydroxyl c xc nh dng bi s ph hp hon ton v s liu ph 1H
v 13C-NMR ti cc v tr tng ng ca SM3 vi cc hp cht c cu trc tng
t [177].

Hnh III.1.4.3.h. Ph ESI-MS ca SM3

183

T tt c cc phn tch nu, cu trc ha hc ca SM3 c xc nh l

1E,3E,7E,11E-cembratetraene-15R(),16-diol. y l mt hp cht c cu trc
mi v c chng ti t tn l sarcophytol V.
III.1.4.4. Hp cht SM4: sarcophytol X
Hp cht SM4 cng c phn lp di dng cht du khng mu. Bng
cc d kin ph khi lng (positive ESI-MS m/z: m/z 334.8 [M-H2O+H]+, 316.8
[M-2H2O+H]+, 298.8 [M-3H2O+H]+) v ph cng hng t ht nhn (xem Bng
III.1.4.4) cho php xc nh cng thc phn t ca n l C20H32O5.

Hnh III.1.4.4.a. Ph 1H-NMR ca SM4

18

O
7
3

1
11

OH

16

19

14

15

OH

OH
17

20

Hnh III.1.4.4.b. Ph cu trc ha hc ca SM4

Trn ph 1H-NMR ca SM4 xut hin cc tn hiu c trng ca hai
proton olefin cng hng ti 6,49 (1H, d, J = 1.0 Hz, H-3) v 4,98 (1H, dt, J =
7,0 v 1,0 Hz, H-11); mt nhm oximetin v mt nhm oximetilen tng ng ti
2,87 (1H, dd, J = 8,0 v 3,5 Hz, H-7) v 3,48 (1H, dd, J = 13,0 v 7,0 Hz, Ha16)/ 3,43 (1H, dd, J = 13,0 v 7,0 Hz, Hb-16). S xut hin cu bn nhm metyl
c khng nh bi cc tn hiu cng hng ti 1,44 (3H, s, H-17), 2,11 (3H,
d, J = 1,0 Hz, H-18), 1,24 (3H, s, H-19) v 1,55 (3H, d, J = 1,0 Hz, H-20).

184

Hnh III.1.4.4.c. Ph 13C-NMR v cc ph DEPT ca SM4

Bng III.1.4.4. Kt qu ph NMR ca SM4

C

Ca, b

1
2
3
4
5
6
7
8

71,95
199,42
122,86
157,89
38,44
25,59
61,77
60,87

Ha, c
multiplicity (J in Hz)
6,49 d (1,0)
2,35 m
1,70 m/2,02 m
2,87 dd (8,0, 3,5)
185

HMBC
(H to C)

2, 5, 18
4, 6, 7
5, 7, 8
6

9
10
11
12
13
14
15
16

37,11
22,82
126,57
134,59
34,91
27,41
65,79
65,92

1,46 m/1,97 m
2,08 m
4,98 dt (7,0, 1,0)
2,15 m/2,22 m
1,65 m/ 2,48 m
3,48 dd (13,0, 7,0)
3,43 dd (13,0, 7,0)
17
15,74
1,44 s
18
19,13
2,11 d (1,0)
19
17,98
1,24 s
20
15,25
1,55 d (1,0)
a
o trong CDCl3, b125 MHz, c500 MHz

7, 8, 10, 11
13, 20
1, 12, 14
1, 12, 13
1
1, 15, 16
3, 4, 5
7, 8, 9
11, 12, 13

Ph 13C-NMR ca SM4 xut hin 20 tn hiu cacbon c trng cho mt

hp cht ditecpen dng cembranoit. Trong , s xut hin ca mt nhm
cacbonyl c khng nh bi tn hiu cng hng ti 199,42 (C-2), hai lin kt
i u b th 3 v tr ti 122,86 (CH, C-3)/157,89 (C, C-4) v 126,57 (CH, C11)/134,59 (C, C-12). Cc tn hiu cng hng ti 61,77 (CH, C-7)/60,87 (C, C8) c trng cho s tn ti ca mt cu epoxy [173]. Ngoi ra, s xut hin ca
mt nhm oximetilen v hai cacbon bc bn ni vi nguyn t xy cng c
khng nh tng ng bi cc tn hiu cng hng ti 65,92 (CH2, C-16), 71,95
(C, C-1) v 65,79 (C, C-15). Cc tn hiu ca bn nhm metyl cng hng ti
15,74 (C-17), 19,13 (C-18), 17,98 (C-19) v 15,25 (C-20). Thng qua ph HSQC,
tt c cc tn hiu cacbon c gn vi cc tn hiu proton tng ng v kt qu
chi tit c a ra trong Bng III.1.4.4.

Hnh III.1.4.4.d. Ph HSQC ca SM4

186

Hnh III.1.4.4.e. Ph HSQC ca SM4

18

O
7
3

OH

16

19

11
14

15

OH

OH
17

20

Hnh III.1.4.4.f. Cc tng tc HMBC (HC) chnh ca SM4

S liu NMR ti tt c cc v tr ca SM4 c gn bng cch so snh vi
cc s liu c cng b ca cc hp cht c cu trc tng t [173-174] v s
phn tch chi tit cc tng tc trn ph HMBC. V tr ca nhm cacbonyl c
khng nh ti C-2 bi s xut hin pic tng tc ca n vi proton H-3. Cc
tng tc HMBC t cc proton H3-17 sang cc cacbon C-1, C-15 v C-16 cng
vi tng tc t proton H-16 sang cacbon C-1 khng nh v tr th ca cc nhm
hydroxyl ti C-1, C-15 v C-16. Bng cc phn tch tng t cho php khu ni
ton b cc v tr trong phn t ca SM4 nh a ra trn Hnh III.1.4.4.b. Cc
phn tch tng tc HMBC chi tit c ch ra Bng III.1.4.4 v Hnh III.1.4.4.f.
T tt c cc phn tch nu phn trn, cu trc ha hc ca SM4 c xc
nh l 7(S),8(S)-epoxy-3E,11E-cembradiene-1,15,16-triol-2-one. y l mt hp
cht mi v c chng ti t tn l sarcophytol X.

187

Hnh III.1.4.4.f. Ph ESI-MS ca SM4

III.1.4.5. Hp cht SM5: (+)-7,8-dihydroxydeepoxysarcophytoxide
Hp cht SM5 cng c phn lp di dng cht du khng mu. Cc
kt qu ph khi lng (positive ESI-MS: m/z 343 [M+Na]+, 303 [M-H2O+H]+)
v ph NMR cho php xc nh cng thc phn t ca n l C20H32O3.

Hnh III.1.4.5.a. Ph 1H-NMR ca SM5

188

18

HO

H
O

HO

16

19
1

15

11
14

17

20

Hnh III.1.4.5.b. Cu trc ha hc ca SM5

Hnh III.1.4.5.c. Ph 13C-NMR v cc ph DEPT ca SM5

189

Trn ph 1H-NMR ca SM5 xut hin cc tn hiu c trng ca hai

proton olefin cng hng ti 5,24 (1H, d, J = 11,0 Hz, H-3) v 5,20 (1H, t, J =
7,0 Hz, H-11); hai proton oximetin v hai proton oximetilen ti 5,40 (1H, d, J =
11,0 Hz, H-2), 3,43 (1H, t, J = 7,0 Hz, H-7) v 4,44 (1H, d, J = 11,0 Hz, Ha-16)/
4,53 (1H, dd, J = 11,0 v 4,0 Hz, Hb-16). Bn nhm metyl c khng nh bi
cc tn hiu proton ti 1,67 (3H, s, H-17), 1,84 (3H, s, H-18), 1,20 (3H, s, H-19)
v 1,64 (3H, s, H-20).
Bng III.1.4.5. Kt qu ph NMR ca SM5
C

C * [168]

C # [168]

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

133,4
84,0
127,3
138,8
35,61
28,0
72,7
78,8
39,5
24,62
123,0
136,3
36,51
24,92
128,0
78,5

133,4
83,9
126,6
139,1
35,54
26,65
72,8
75,5
36,64
25,25
124,2
135,6
36,94
23,65
127,5
78,2

133,90
85,14
125,16
141,72
36,30
29,82
74,39
75,44
38,32
22,78
125,20
136,59
37,50
23,83
128,20
78,41

17
18
19
20

10,3
15,63
26,6
15,93

10,1
15,26
24,2
15,86

10,51
17,63
24,65
16,04

Ha, c
mult. (J in Hz)
5,40 d (11,0)
5,24 d (11,0)
2,24 m
1,80 m/1,61 m
3,43 t (8,0)
1,77 m/1,67 m
2,21 m/2,11 m
5,20 t (7,0)
2,17 m/2,05 m
2,20 m/1,93 m
4,44 d (11,0)
4,53 dd (11,0, 4,0)
1,67 s
1,84 s
1,20 s
1,64 s

HMBC
(HC)

1, 2, 5
6, 18

10
9, 11
13
11, 12, 14, 20
1, 2, 13, 15
1, 15
1, 15, 16
3, 4, 5
7, 8, 9
11, 12, 13

o trong CDCl3, b125 MHz, c500 MHz,1-6ccgi tr c cng s c th i ch cho nhau, *gi tr ph ca
(+)-7,8-dihydroxydeepoxysarcophytoxide, # gi tr ph ca (+)-7,8-dihydroxydeepoxysarcophytoxide.

Ph 13C-NMR ca SM5 xut hin 20 tn hiu cacbon c trng ca mt

hp cht ditecpen dng cembranoit c hai lin kt i b th 3 v tr, mt lin kt
i b th hon ton, hai cacbon oxitmetin, mt cacbon oximetilen, mt cacbon
bc bn ni vi nguyn t xy v bn nhm metyl.

190

Hnh III.1.4.5.d. Ph HSQC ca SM5

Hnh III.1.4.5.e. Ph HMBC ca SM5

191

18

HO

H
O

HO

16

19
1

15

11
14

17

20

Hnh III.1.4.5.f. Cc tng tc HMBC (HC) chnh ca SM5

Cc s liu ph 13C-NMR ca SM5 c gn vi cc s liu proton tng
ng thng qua ph HSQC. Cc s liu ph ny hon ton tng t nh s liu ph
ca SM1. S khc bit duy nht gia hai hp cht ny l s mt i nhm metoxy
trong phn t ca SM5 so vi SM1. T kt qu ph khi lng cho php khng
nh SM5 l dn xut 7,8-dihydroxy ca SM1. So snh k lng s liu ph 13CNMR ca SM5 vi cc hp cht c cng cu trc phng (Bng III.1.4.5) [168]
cho php xc nh cu hnh ca 2 nhm hydroxy ti v tr C-7 v C-8 u l dng
. iu ny mt ln na c khng nh bi cc tng tc khng gian gia
proton H-7 vi cc proton H3-19 trn ph ROESY. Tng tc ROESY gia proton
H-2 v cc proton H3-18 cho php xc dnh cu hnh ti v tr C-2 l dng S.

Hnh III.1.4.5.g. Ph ROESY ca SM5

192

T tt c cc phn tch nu trn cho php xc nh cu trc ha hc ca

SM5 l (+)-7,8-dihydroxydeepoxysarcophytoxide. y l mt hp cht c cu
trc mi ln u tin c phn lp t thin nhin.

Hnh III.1.4.5.h. Ph ESI-MS ca SM5

III.1.4.6. Hp cht SM6: (+)-Sarcophine
Hp cht SM6 cng c phn lp di dng cht du khng mu. Cng
thc phn t ca n c xc nh l C20H28O3 bng cc kt qu ph khi lng
(positive ESI-MS: m/z 339.1 [M+Na]+, 317.0 [M+H]+, 299.0 [M-H2O+H]+) v
ph NMR.

Hnh III.1.4.6.a. Ph 1H-NMR ca SM6

193

18

16

19
1
11

14

15
17

20

Hnh III.1.4.6.b. Cu trc ha hc ca SM6

Trn ph 1H-NMR ca SM6 xut hin cc tn hiu c trng ca hai
proton olefin ti 4,99 (1H, d, J = 9,0 Hz, H-3) v 5,11 (1H, t, J = 7,0 Hz, H11); hai proton oxymetin ti 5,45 (1H, d, J = 9,0 Hz, H-3) v 2,66 (1H, t, J =
5,5 Hz, H-7); v bn nhm metyl ti 1,86 (3H, s, H-17), 1,93 (3H, s, H-18),
1,27 (3H, s, H-19) v 1,61 (3H, s, H-20).
Bng III.1.4.6. Kt qu ph NMR ca SM6
C

C*[173]

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20

162,2
78,8
120,6
144,0
37,4
25,2
61,5
59,9
39,0
23,3
124,9
135,5
36,4
27,5
122,9
175,0
8,9
16,1
17,7
15,5

162,78
79,83
119,22
144,88
38,37
25,76
61,77
60,07
36,88
22,97
124,69
134,47
35,13
26,79
123,18
174,89
9,18
16,74
18,09
15,52

Ha, c
multiplicity (J in Hz)
5,45 d (9,0)
4,99 d (9,0)
2,31 m/2,22 m
1,83 m/1,72 m
2,66 t (5,5)
2,05 m/1,19 dt (13,0, 4,0)
2,21 m/1,94 m
5,11 t (7,0)
2,34 m/1,92 m
2,36 m/1,71 m
1,86 s
1,93 s
1,27 s
1,61 s

o trong CDCl3, b125 MHz, c500 MHz, *s liu ph ca (+)-sarcophine.

194

Hnh III.1.4.6.c. Ph 13C-NMR ca SM6

Hnh III.1.4.6.d. Ph 13C-NMR v cc ph DEPTca SM6

Ph 13C-NMR ca SM6 xut hin 20 tn hiu cacbon c trng cho mt
hp cht ditecpen dng cembranoit. Trong c: hai lin kt i b th 3 v tr,
mt lin kt i b th hon ton, mt cu epoxy v bn nhm metyl. S xut
hin ca mt nhm oximetin ti 79,83 (C-2) v mt cacbon bc bn ti
174,89 (C-16) gi cho s c mt ca mt hp cht furanocembranoit.

195

Hnh III.1.4.6.e. Ph ESI-MS ca SM6

Cc d kin nu trn gi ti hp cht sarcophine, mt furanocembranoit
c trng ca chi Sarcophyton. T , s liu ph 13C-NMR c so snh trc
tip vi s liu c cng b ca hp cht ny [173]. S ph hp hon ton ti
cc v tr tng ng cho php khng nh cu trc ha hc ca SM6 l (+)sarcohine.
Theo cc ti liu cng b, hp cht ny l mt trong nhng thnh phn
quan trng trong h thng phng v ha hc ca cc loi san h mm chng
li k th. Ngoi ra, n cn th hin hot tnh gy c t bo cao i vi nhiu
dng t bo nh dng A549 (ED50 = 24,66 g/ml), HT-29 (ED50 = 22,88 g/ml)
v c bit vi dng P-388 (ED50 = 3,27 g/ml) [179]. Khng nhng th (+)sarcophine cn th hin kh nng chng s di tr ca cc t bo ung th rt cao
[s di tr (migration) v tng sinh (proliferation) l cc yu t to nn qu trnh di
cn (metastasis) ca bnh ung th], kh nng ny khng phi do tnh c t bo
to nn m l do n c ch cc c ch di tr ca cc t bo ung th [180]. Mt
iu th v na l, hp cht ny cn cn th hin hot tnh chng xy ha, c
nh gi bng kh nng c ch s sn sinh ion m peoxit (O2-) v thromboxane B2
(TXB2) bi cc t bo tiu thn kinh m no chut s sinh c hot ha
vi lipopolysaccharit ca E. coli (LPS) [181] khi c kch thch bng phorbol
12-myristate 13-acetate (PMA)
III.1.4.7. Hp cht SM7: sarcophytoxide
Hp cht SM7 cng c phn lp di dng cht du khng mu. Bng
cc kt qu ph khi lng (positive ESI-MS: m/z 325.0 [M+Na]+, 303.0 [M+H]+,
285.0 [M-H2O+H]+) v ph NMR cho php xc nh cng thc phn t ca n l
C20H30O2.
196

18

16

19
1
11

14

15
17

20

Hnh III.1.4.7.a. Cu trc ha hc ca SM7

Hnh III.1.4.7.b. Ph 1H-NMR ca SM7

Trn ph 1H-NMR ca SM7 xut hin cc tn hiu c trng ca hai
proton olefin, hai nhm oximetin, mt nhm oximetilen v bn nhm metyl. Ph
13
C-NMR ca SM7 xut hin 20 tn hiu cacbon ca mt hp cht ditecpen dng
cembranoit. Trong c c cc tn hiu c trng ca: hai lin kt i b th 3 v
tr, mt lin kt i b th hon ton, hai nhm oximetin, mt nhm oximetilen,
mt cacbon bc bn ni vi nguyn t xy v 4 nhm metyl. Tt c cc s liu
ph cacbon c gn vi cc proton tng ng thng qua ph HSQC.

197

Hnh III.1.4.7.c. Ph 13C-NMR ca SM7

Bng III.1.4.7. Kt qu ph NMR ca SM7
C

C*[178]

Ca, b

Ha, c
multiplicity (J in Hz)

HMBC
(H to C)

1
133,9
133,90
2
85,3
85,32
5,41 m
3
125,5
125,41
5,27 d (10,0)
1, 2, 5
4
140,2
140,41
5
35,1
35,04
2,33 m/2,24 m
3, 4
6
27,4
27,42
1,81 m/1,71 m
7
62,3
62,44
2,70 t (5,5)
5, 6, 9
8
60,1
60,21
9
38,8
38,95
2,07 m/1,15 dt (13,0, 4,0)
10
10
23,1
23,18
2,21 m/1,96 m
11
123,5
123,39
5,13 t (7,5)
10, 13, 20
12
135,8
135,95
13
38,4
38,57
2,25 m/1,93 m
11, 12, 14, 20
14
23,7
23,76
2,35 m/1,66 m
1, 2
15
128,1
128,14
16
78,3
78,35
4,46 /4,53
1, 15
17
10,3
10,41
1,66 s
1, 15, 16
18
17,6
17,71
1,86 s
3, 4, 5
19
16,4
16,42
1,27 s
7, 8, 9
20
15,4
15,45
1,58 s
11, 12, 13
a
b
c
*
o trong CDCl3, 125 MHz, 500 MHz, s liu ph ca sarcophytoxide.

198

Hnh III.1.4.7.d. Ph 13C-NMR v cc ph DEPT ca SM7

Hnh III.1.4.7.e. Ph HSQC ca SM7

199

Hnh III.1.4.7.f. Ph HMBC ca SM7

Cc d liu nu gi ti hp cht sarcophytoxide, mt cembranoit
ditecpen kh ph bin trong cc loi san h mm. Tt c cc gi tr ph NMR ca
SM7 c gn bng cch so snh vi cc s liu c cng b ca hp cht
sarcophytoxide v c khng nh li bi cc tng tc trn ph HMBC (Bng
III.1.4.7 v hnh III.1.4.7.f-h).
18

16

19
1

15

11
14

17

20

Hnh III.1.4.7.g. Cc tng tc HMBC (HC) chnh ca SM7

T cc d kin nu trn, cng vi s ph hp hon ton v s liu ph 13CNMR so vi cc s liu c cng b (Bng III.1.4.7) [178] cho php xc nh
cu hnh ti v tr C-2 l R, v tr C-7 v C-8 l dng S, v cu trc ha hc ca
SM7 l sarcophytoxide.

200

Hnh III.1.4.7.h. Ph ESI-MS ca SM7

III.1.4.8. Hp cht SM8: Sarcomilasterol
Hp cht SM8 c phn lp di dng tinh th hnh kim mu trng, im
chy 160-161 oC. Cng thc phn t ca n c xc nh l C28H48O4 bng cc
kt qu ph khi lng (positive ESI-MS m/z: 471.0 [M+Na]+, 449.1 [M+H]+,
431.1 [M-H2O+H]+) v ph cng hng t ht nhn (NMR). T cng thc phn
t cho php xc nh h s khng no trong phn t ca SM8 l 5.

Hnh III.1.4.8.a. Ph 1H-NMR ca SM8

201

28

21

26
24

18

11

HOH2C
19

20
17

25

27

13
14

1
8

10
3

HO

5
6

OH

Hnh III.1.4.8.b. Cu trc ha hc ca SM8

Ph 1H-NMR c dng c trng ca mt hp cht steroit vi cc tn hiu
c trng ca su nhm metyl cng hng ti 0,65 (3H, s, H-18), 1,12 (3H, s,
H-19), 0,97 (3H, d, J = 7,0 Hz, H-21), 0,79 (3H, d, J = 7,0 Hz, H-26), 0,86 (3H,
d, J = 7,0 Hz, H-27) v 0,78 (3H, d, J = 7,0 Hz, H-28); hai nhm oximetin v
mt nhm oximetilen ti 3,53 (1H, m, H-3), 4,21 (1H, dd, J = 10,0 v 2,2 Hz,
H-6) v 3,82 (1H, m, Ha-11)/3,63 (1H, m, Hb-11); v mt proton olefin ti 6,61
(1H, d, J = 2,2 Hz, H-8).

Hnh III.1.4.8.c. Ph 13C-NMR ca SM8

202

Hnh III.1.4.8.d. Ph 13C-NMR v cc ph DEPT ca SM8

Bng III.1.4.8. Kt qu ph NMR ca SM8
Ha, c multiplicity (J in Hz)

HMBC (H C)

31,95
30,21
69,72
32,48
48,54
68,80
148,57
136,63
206,08
44,99
59,06
40,89
46,14
42,75

1,84 m/1,42 m
1,87 m/1,42 m
3,53 m
2,24 m/1,44 m
1,75 m
4,21 dd (10,0, 2,2)
6,61 d (2,2)
3,82 m /3,63 m
1,60 m /1,08 m
3,34 t (10,0)

3
3

26,60
26,27
49,70
17,42
16,13
35,34
18,96
33,14
31,40
39,14
31,58
17,67
20,51
15,49

1,58 m /1,40 m
1,82 m/1,58 m
1,67 m
0,65 s
1,12 s
1,36 m
0,97 d (7,0)
1,40 m /0,93 m
1,42 m /0,93 m
1,18 m
1,55 m
0,79 d (7,0)
0,86 d (7,0)
0,78 d (7,0)

C [182]

C a, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14

33,2
31,3
70,8
33,7
49,9
69,7
149,3
137,4
206,1
46,1
59,2
42,2
47,2

15
16
17
18
19
20
21
22
23
24
25
26
27
28

28,0
27,2
51,6
17,7
16,5
36,0
19,4
36,6
25,6
40,7
29,2
23,1
22,9
-

43,6

o trong CDCl3+CD3OD, b125 MHz, c500 MHz,

203

6
4, 5, 7, 8
5, 9, 14

11, 13, 14, 17

7, 8, 9, 12, 13, 15,
16, 18

12, 13, 14, 17

1, 5, 9, 10
17, 20, 22

24, 25, 27
24, 25, 26
23, 24, 25

Ph 13C-NMR ca SM8 xut hin 28 tn hiu cacbon dng steroit. Trong

xut hin cc tn hiu c trng ca mt lin kt i b th 3 v tr ti 148,57
(C-7)/136,63 (C-8); hai nhm oximetin v mt nhm oximetilen tng ng ti
69,72 (C-3), 68,80 (C-6) v 59,06 (C-11); v su nhm metyl ti 17,42 (C-18),
16,13 (C-19), 18,96 (C-21), 17,67 (C-26), 20,51 (C-27) v 15,49 (C-28). Cc
tng tc trn ph HSQC cho php gn tt c cc gi tr ph cacbon vi gi tr
ph proton tng ng. Ngoi ra, s xut hin tn hiu cacbon ti 206,08 (C-9)
khng nh s tn ti ca mt nhm ktn nm v tr lin hp vi lin kt i
c minh chng bi s chn mnh m ca proton olefin ( 6,61, H-7) nm v
tr so vi nhm ktn.

Hnh III.1.4.8.e. Ph HSQC ca SM8

Hnh III.1.4.8.f. Ph HMBC ca SM8

204

Vi mc khng no trong phn t ca SM8 l 5, cng vi s xut hin

ca mt nhm ktn, mt lin kt i cho php xc nh n phi c cu trc 3
vng v do c khng nh l mt secosterol. C th l cu trc 9,11secosterol c cha nhm hydroxyl v tr C-11 c xc nh t cc tn hiu
cng hng c trng ca h proton tng tc c lp cha cc tn hiu cng
hng tng tc ln nhau ti H 1,60 (Ha-12), 1,08 (Hb-12), 3,82 (Ha-11), v 3,63
(Hb-11), c gn tng ng vi 2 tn hiu cacbon cng hng ti 59,06 (CH2,
C-11) and 40,89 (CH2, C-12) thng qua ph HSQC. C s liu chuyn dch
ha hc 1H v 13C ca on etyl ny hon ton trng khp vi cc gi tr tng
ng trong cc hp cht 9,11-secosterol c cu trc tng t c cng b
[183-185]. Cu trc 7,11-hydroxyl, mch nhnh v cu trc ton phn ca SM8
c xc nh bng cch so snh s liu ph NMR vi cc s liu tng ng
c cng b [182, 186] v c khng nh li bi cc tng tc xa trn ph
HMBC (Bng III.1.4.8 v Hnh III.1.4.8.i).

Hnh III.1.4.8.g. Ph ROESY ca SM8

CH3

HO

CH3
CH3

H O
H

HO
OH
H

CH3

Hnh III.1.4.8.h. Cc tng tc ROESY quan trng ca SM8

205

28

26

21
24
18

20
17

25

27

13

HOH2C 11
O
19

14

1
8

10
3

HO

5
6

OH

Hnh III.1.4.8.i. Cc tng tc HMBC (HC) chnh ca SM8

Cu hnh ti cc tm bt i trong phn t ca SM8 c xc nh bng
cch so snh s liu ph NMR vi cc s liu c cng b ca cc hp cht
9,11-secosterol c cng b v c khng nh li bi cc tng tc khng
gian trn ph ROESY. Tn hiu cng hng ti 3,53 (1H, m) c trng cho proton
H-3 vi cu hnh ca cc hp cht steroit [187]. Cu hnh ca nhm hydroxyl
ti v tr C-6 d dng c xc nh da trn gi tr chuyn dch ha hc ca
cacbon C-6 ti 68,80 (ti 66,0 cho cu hnh ) [182, 188].

Hnh III.1.4.8.j. Ph ESI-MS ca SM8

Trn ph ROESY, tn hiu tng tc khng gian gia proton H-3 v H-5
th hin cu hnh trans ca cc vng A v B; tn hiu tng tc gia H-6 v H319, cng vi hng s tng tc J = 10,0 Hz gia H-6 v H-5 v s tng mnh
chuyn dch ha hc ca H-4eq ( 2,24), mt ln na chng minh cu hnh ca
nhm hydroxyl ti v tr C-6. Cui cng, cu hnh ca vng D c xc nh bi
cc tng tc nhn c gia H-14 ( 3,34) v H-17 ( 1,67), Hb-12 ( 1,08), Ha206

11 ( 3,82) v Hb-11 ( 3,63). T tt c cc phn tch nu trn, cu trc ha hc

ca SM8 c xc nh l 24R-methyl-3,6,11-trihydroxy-9,11-seco-5cholest-7-ene-9-one. y l mt hp cht c cu trc mi v c chng ti t
tn l sarcomilasterol.
III.1.4.9. Hp cht SM9: sarcoaldesterol B
Hp cht SM9 nhn c di dng cht bt mu trng. Cng thc phn
t ca n c xc nh l C28H50O4 bng cc d kin ph khi lng (positive
ESI-MS: m/z 473,1 [M + Na] +, 433,1 [M - H2O + H] +, 415,1 [M - 2H2O + H] +,
397,1 [M - 3H2O + H] +) v ph cng hng t ht nhn.

Hnh III.1.4.9.a. Ph 1H-NMR ca SM9

28

21

26
24

18

HO
19

20
17

13

11

25

27

9
10

HO
OH
OH

Hnh III.1.4.9.b. Cu tr ha hc ca SM9

Ph 1H-NMR ca SM9 c trng cho mt hp cht steroit. Trong , s
xut hin ca ba nhm oximetin c khng nh bi cc tn hiu cng hng ti

207

 3,84 (1H, m, H-3), 3,46 (1H, br s, H-6) v 4,01 (1H, m, H-11). Ngoi ra cc tn
hiu cng hng ti 0,75 (3H, s, H-18), 1,29 (3H, s, H-19), 0,99 (3H, d, J = 7,0
Hz, H-21), 0,83 (3H, d, J = 7,0 Hz, H-26), 0,90 (3H, d, J = 7,0 Hz, H-27) v 0,82
(3H, d, J = 7,0 Hz, H-28) khng nh s tn ti ca nhm metyl v gi cho cu
trc nhnh bn dng ergosterol tng t nh ca SM8.

Hnh III.1.4.9.c. Ph 13C-NMR ca SM9

Bng III.1.4.9. Kt qu ph NMR ca SM9
C

C [189]

1
2
3
4
5
6
7
8
9
10
11
12

35,5
33,0
67,5
43,3
76,9
76,5
35,8
30,1
53,0
41,1
68,6
52,9

H a, c

HMBC

multiplicity (J in Hz)

(H C)

a, b

35,32
32,03
69,43
41,91
77,45
76,55
35,15
30,31
52,99
40,91
68,17
52,75

2,08 m/1,81 m
1,77 m/1,52 m
3,84 m
2,09 m/1,57 m
3,46 br s
1,81 m/1,52 m
1,80 m
1,47 m
4,01 m
2,28 dd m/1,26 m
208

5, 8, 10

10
9, 11, 13, 14

13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
a,

43,6
56,0
24,8
28,7
56,5
13,5
17,8
36,6
19,1
34,0
31,0
39,4
31,8
17,8
20,8
15,7

44,25
56,53
25,26
29,36
57,44
13,40
17,56
37,46
19,39
34,83
31,70
40,34
32,66
18,12
20,96
16,00

1,21 m
1,62 m/1,15 m
1,90 m/1,32 m
1,21 m
0,75 s
1,29 s
1,40 m
0,99 d (7,0)
1,47 m/1,01 m
1,45 m/1,01 m
1,23 m
1,60 m
0,83 d (7,0)
0,90 d (7,0)
0,82 d (7,0)

12, 13, 14, 17

1, 5, 9, 10
17, 20, 22

26, 27
24, 25, 27
24, 25, 26
23, 24, 25

o trong CD3OD, b125 MHz, c500 MHz, *s liu ph ca sarcoaldesterol.

Hnh III.1.4.9.d. Ph 13C-NMR v cc ph DEPT ca SM9

Ph 13C-NMR ca SM9 xut hin cc tn hiu c trng cho mt hp cht
steroit c 28 cacbon. Trong s xut hin tn hiu ca 6 nhm metyl ti 13,40
(C-18), 17,56 (-19), 19,39 (C-21), 18,12 (C-26), 20,96 (C-27) v 16,00 (C-28)
khng nh s tn ti ca nhnh bn dng ergosterol tng t nh SM8. Ngoi
ra, s xut hin ca ba nhm oximetin v mt cacbon bc bn ni vi nguyn t
xy cng c khng nh tng ng bi cc tn hiu cng hng ti 69,43 (C3), 76,55 (C-6), 68,17 (C-11) v 77,45 (C-5). Cc tn hiu cacbon c gn vi
cc tn hiu proton tng ng thng qua ph HSQC (Bng III.1.4.9).
209

Hnh III.1.4.9.e. Ph HSQC ca SM9

Nh vy c th s b xc nh, SM9 l mt steroit dng ergosterol cha 3
nhm oximetin v mt cacbon bc bn ni vi nguyn t xy. Nhm -hydoxyl
ti v tr C-3 d dng c xc nh bi tn hiu proton c trng ti 3,84 (1H, m,
H-3) [187]. xc nh v tr ca cc nhm hydroxyl cn li, ph cng hng t
ht nhn hai chiu tng tc xa HMBC c o. Tn hiu tng tc HMBC t
H3-19 sang C-5 khng nh v tr ca nhm hydroxyl ti C-5. Nhm hydroxyl C-6
c xc nh bi cc tn hiu tng tc HMBC t proton ca n sang cacbon C5, C-8 v C-10. Cui cng cc tn hiu tng tc t proton H-12 sang C-11 v t
proton H-11 sang C-10 khng nh v tr ca nhm hydroxyl ti C-11.

Hnh III.1.4.9.f. Ph HSQC ca SM9

210

28

21

HO
19

26
20

18

17

25

27

13

11

9
10

HO
OH
OH

Hnh III.1.4.9.g. Cc tng tc HMBC (CH) chnh ca SM9

T tt c cc phn tch nu trn v s ph hp hon ton v s liu ph
NMR ca SM9 vi cc s liu c cng b [189], cho php khng nh cu
trc ha hc ca n l ergosta-3,5,6,11-tetraol hay cn c gi l
sarcoaldesterol B. Hp cht ny c phn lp t loi Sarcophyton sp. Tuy
nhin y l ln u tin n c phn lp t loi S. mililatensis.

Hnh III.1.4.9.h. Ph ESI-MS ca SM9

III.1.4.10. Hp cht SM10: ergosta-1,3,5,6-tetraol
Hp cht SM10 nhn c di dng tinh th hnh kim mu trng. Cng
thc phn t ca n c xc nh trng vi cng thc phn t ca SM9 bng cc
d kin ph khi lng (positive ESI-MS: m/z 473,1 [M + Na] +, 450,9 [M + H]+)
v ph cng hng t ht nhn.

211

Hnh III.1.4.1.a. Ph 1H-NMR ca SM10

28

21

26
24

18

20
17

OH
19

13

11

25

27

9
10

HO
OH
OH

Hnh III.1.4.1.b. Cu trc ha hc ca SM10

S liu ph 1H v 13C-NMR ca SM10 hon ton tng t nh s liu ca
SM9. S thay i d nhn thy nht l tn hiu cacbon oximetin C-11 trong phn
t ca SM9 ti 68,17 b dch chuyn ln vng trng thp ti 74,33 trong
phn t ca SM10. iu ny cho php khng nh nhm oximetin C-11 b
thay i v tr. khng nh v tr ca nhm hydroxyl ny, ph HMBC c
o. Tn hiu tng tc HMBC gia proton H3-19 v cacbon C-1 ( 74,33) cho
php khng nh v tr th ca nhm hydroxyl C-1.

212

Hnh III.1.4.1.c. Ph 13C-NMR v cc ph DEPT ca SM10

Bng III.1.4.10. Kt qu ph NMR ca SM10
C

C* [190]

C a, b

1
2
3
4
5
6

73,7
44,1
65,3
43,2
76,9
77,0

74,33
42,48
65,98
41,55
77,52
77,10

H a, c
mult. (J in Hz)
3,97 dd (10,5, 4,5)
2,01 m/1,57 m
4,03 m
2,07 dd (12,5, 11,5)
3,45 br s

213

HMBC
(H C)
9, 10, 19
1, 3, 4, 10
3, 5, 6
4, 5, 8, 10

7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
a,

35,7
31,7
47,1
44,9
25,0
41,3
42,6
56,7
24,9
28,5
56,7
12,5
10,7
34,0
19,0
36,6
31,1
39,3
31,9
17,8
20,7
15,7

34,95
32,12
47,38
44,89
25,02
41,99
43,35
57,55
25,57
29,13
57,76
12,61
10,17
37,58
19,37
35,21
31,74
40,45
32,71
18,07
20,88
15,95

1,48 m/1,00 m
1,77 m
1,65 m
2,15 m/1,42 m
2,01 m/1,21 m
1,15 m
1,61 m/1,17 m
1,86 m/1,29 m
1,15 m
0,73 s
1,15 s
1,40 m
0,95 d (7,0)
1,73 m/1,54 m
1,45 m/1,00 m
1,23 m
1,61 m
0,84 d (7,0)
0,90 d (7,0)
0,83 d (7,0)

recorded in CD3OD, b125 MHz, c500 MHz, *s liu ph ergosta-1,3,5,6-tetraol

Hnh III.1.4.1.d. Ph HSQC ca SM10

214

11, 13
16
12, 13, 14, 17
1, 5, 9, 10
17, 20, 22

24, 25, 27
24, 25, 26
23, 24, 25

Hnh III.1.4.1.e. Ph HMBCca SM10

28

21

26
20

18

17

OH

19

25

27

13

11

1
9

HO
OH
OH

Hnh III.1.4.10.f. Cc tng tc HMBC (HC) chnh ca SM10

Tt c cc phn tch nu cng vi s ph hp hon ton v s liu ph
NMR ti cc v tr tng ng ca SM10 so vi cc s liu cng b [190-191]
cho php khng nh cu trc ha hc ca n l ergosta-1,3,5,6-tetraol. Hp
cht ny c phn lp t cc loi san h mm S. glaucum v Lobophytum sp.
Tuy nhin y l ln u tin n c phn lp loi S. mililatensis.

215

Hnh III.1.4.10.g. Ph ESI-MS ca SM10

III.1.5. Kt qu nh gi hot tnh sinh hc ca cc hp cht
III.1.5.1. Kt qu hot tnh chng long xng thng qua cc c ch kch
thch s bit ha ca nguyn bo xng MC3T3-E1.
Nh bit long xng c nhn bit bi s gim khi lng xng, lm
tng tnh dn v nguy c gy xng. Rt nhiu bnh nhn mc bnh long xng
mt mt lng xng tng i ln. V vy rt cn pht trin mt phng thc
tng cng lng xng cho c th thng qua cc kch thch to xng mi.
S thiu ht estrogen cng l mt nguyn nhn gy nn bnh long xng, c
bit l cc ph n sau 65 tui. Estrogen c tc dng lm gim s lng cc t
bo hy xng v tng cng qu trnh hnh thnh cc nguyn bo xng, lm
chm qu trnh bin i ca xng v vy m ngn nga c qu trnh mt khi
lng xng trong c th. Tuy nhin, estrogen cng thng gy nn nhng tc
dng khng mong mun nh hnh thnh v pht trin cc khi u ph thuc
estrogen nh cc khi u c t cung v u v [192]. iu ny thc y mnh m
nhng nghin cu nhm tm kim cc hp cht c hiu lc nh esgrogen ln cc
v tr chn lc nh b khung xng v h tim mch m khng gy nn cc hiu
ng ph. Cc tc nhn iu bin chn lc ER (SERM) c hot ng tng t nh
cc estrogen nhng li khng gy tc dng ph v v t cung [193-194].
Nhm thm d kh nng ca cc mu th nghim i vi s kch thch cc
chc nng hot ng ca nguyn bo xng, cc th nghim v nh hng ca
mu th i vi s sinh trng ca t bo, hot lc ca enzim ALP (Alkaline
Phosphatase), Hm lng collagen v v s tch t canxi i vi t bo MC3T3216

E1, mt tin nguyn bo xng, c chp nhn rng ri lm m hnh in vitro

nghin cu s bit ha ca nguyn bo xng.
Cc sterol phn lp c t S. mililatensis c cho vi t bo MC3T3E1 nng 0,3 v 3 M quan st qu trnh sinh trng ca t bo. nng
3 M, s pht trin ca cc t bo ny l rt r rt (hnh III.1.5.1).

Cell growth (%)

130

Compound 1
Compound 2
Compound 3

120

*
*

110
100
90

3
0.

t.
C
on

t.

0.

C
on

3
0.

C
on

t.

80

M
Hnh III.1.5.1. Hiu qu ca cc sterol i vi s pht trin ca t bo MC3T3-E1 cells.
S liu c biu din di dng phn trm vi i chng. Gi tr ca i chng trong
th nghim ny l 0,6430,014 OD. * P<0,05 so vi i chng. Compound 1:
sarcomilasterol, Compound 2: sarcoaldesterol B, Compound 3: ergosta-1,3,5,6tetraol.

Da trn cc thu nhn ban u, chng ti th nghim hot ng gy bit

ha trn dng MC3T3-E1 thng qua cc nh gi v hot ng ca enzim ALP
ni bo v s cung cp canxi. nh hng ca cc sterol trn s bit ha nguyn
bo xng c ghi nhn thng qua nhng nh gi v hot ng ca enzim
ALP, mt trong nhng ch th cho qu trnh bit ha nguyn bo xng. C ba
hp cht trn u c hiu qu lm tng r rt hot ng ca enzim ny nng
0,3 v 3 M. Trong ergosta-1,3,5,6-tetraol (SM10) cho hiu qu mnh
nht. Vi nng 0,3 M, ergosta-1,3,5,6-tetraol kch thch tng cng hot
ng ca enzim ALP n 132,9% khi em so snh vi hot ng ca i chng.
Tip theo , cc th nghim v hiu qu tc ng ca cc sterol i vi s
khong ha, mt qu trnh quan trng trong qu trnh bit ha, c tin hnh
da trn php o s cung cp canxi bng phng php nhum Alizazin . Kt
qu cho thy, hp cht Sarcomilasterol (SM8) v ergosta-1,3,5,6-tetraol c
tc dng kch thch r rt i vi s khong ha. Kt qu ny cho thy cc sterol
t S. mililatensis c hot ng nh mt SERM v iu bin qu trnh ri lon
xng ph n thi k mn kinh. Cc hp cht ny c phn lp vi hm lng
kh cao, iu ny gi m cho nhng nghin cu su hn v cc hp cht ny theo
hng ng dng trong vic phng v iu tr bnh long xng, mt cn bnh rt
ph bin hin nay (c bit l trong cng ng ngi cao tui).
217

Bng III.1.5.1: nh hng ca cc cht ln s bit ha ca t bo MC3T3-E1

Hp cht

SM1

SM2

SM3

SM5

SM6

SM7

SM8

SM9

SM10

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong ha
(%)

0,3
3
30
0,3
3
30
0,3
3
30
0,3
3
30
0,3
3
30
0,3
3
30
0,3
3
30
0,3
3
30
0,3
3
30

127,81,392*
125,12,760*
115,56,288
131,06,834*
127,21,292*
130,83,774*
127,02,449*
121,02,237*
113,22,973*
128,92,660*
139,52,237*
123,41,814*
133,32,091*
130,26,923*
121,91,062*
127,82,995*
125,13,296*
117,42,995*
119,15,214*
118,01,486*
86,410,212*
126,31,486*
124,80,368*
88,542,872*
132,96,623*
126,54,246*
91,300,766*

108,21,860*
101,92,680
91,483,231
101,03,154
102,51,832
111,41,208*
102,70,137
101,01,938
110,41,440*
109,51,898*
106,84,880
99,794,012
103,50,836
109,72,000*
104,01,716
103,23,421
106,91,636*
109,11,734*
106,51,822
104,01,254
69,10,495*
109,72,302*
108,01,012*
98,001,801
105,61,259*
100,03,690
89,634,532

110,62,470*
106,70,403*
105,12,619
97,111,485
101,72,556
107,93,767
96,771,650
97,721,546
101,60,990
100,71,684
104,01,277
105,42,868
96,171,540
99,870,941
99,661,515
96,102,443
97,783,635
102,81,296
101,11,8564
107,21,680*
89,383,609*
100,91,763
104,51,808
97,851,195
104,501,324*
98,123,197
105,82,156

S liu c vit di dng meanSEM, th hin % so vi i chng. Gi tr ca cht i chng cc c

ch th: hot lc ca enzym ALP, thnh phn collagen, v s khong ha tng ng l 1.1220.017 n
v/106 t bo, 26.370.410 g, v 0.4960.004 OD. * P<0.05 so vi i chng.

Trong qu trnh hnh thnh xng, cc nguyn bo xng l t bo ct li

trong qu trnh hnh thnh cht c bn ca xng (bone matrix) v s vi ha. S
to xng bt u bi s sn sinh v tit collagen loi I ca cc nguyn bo
xng, thnh phn to nn 90% cht c bn ca xng, hay m tin ct. Cc
nguyn bo xng c tng hm lng enzym ALP (enzym ct nhm phosphat).
Enzym ALP c gii phng vo m tin ct khi ng qu trnh tch t cc
nguyn t khong. Sau qu trnh khong ha, xng hon chnh ngy cng tr ln
cng v chc vi cc c tnh c hc cn thit chng li cc ngoi lc, nng c
th v bo v cc ni c quan [195].
218

Ngoi cc sterol, cc cembranoit ditecpen cng th hin hot tnh kch thch
mnh m s bit ha nguyn bo xng. c bit, hp cht SM1 tng cng
mnh m c hot lc enzym ALP, s tng hp collagen v s tch t canxi ca t
bo MC3T3-E1 (Bng III.1.5.1). Theo cc hp cht ny c th ng vai tr
quan trng trong vic hnh thnh xng.
III.1.5.2. Hot tnh gy c t bo ca cc hp cht.
Trc tin, tin hnh nh gi gi tr CS% ca cc hp cht. Gi tr CS%
hay chnh l gi tr biu hin lng t bo sng st sau khi cho tip xc vi cc
mu chit c tnh theo cc cng thc trnh by trong phn quy trnh. Cc
cht c CS% < 50 c coi l c hot tnh v s c tip tc tin hnh xc nh
gi tr IC50.
Kt qu th nghim cho thy: Cc hp cht SM1, SM5 v SM8 th hin
hot tnh trn c 3 dng t bo th nghim. Hp cht SM2 v SM6 th hin hot
tnh 2/3 dng t bo th nghim. Hp cht SM3 v SM7 th hin hot tnh
1/3 dng t bo th nghim. Theo cc ti liu cng b, (+)-sarcophine (SM6)
l mt trong nhng thnh phn quan trng trong h thng phng v ha hc ca
cc loi san h mm chng li k th. Ngoi ra, n cn th hin hot tnh gy c
t bo cao i vi nhiu dng t bo nh dng A549 (ED50 = 24,66 g/ml), HT29 (ED50 = 22,88 g/ml) v c bit vi dng P-388 (ED50 = 3,27 g/ml) [179].
Khng nhng th (+)-sarcophine cn th hin kh nng chng s di tr ca cc t
bo ung th rt cao [s di tr (migration) v tng sinh (proliferation) l cc yu t
to nn qu trnh di cn (metastasis) ca bnh ung th], kh nng ny khng phi
do tnh c t bo to nn m l do n c ch cc c ch di tr ca cc t bo ung
th [180]. y l mt du hiu tt, t c s cho cc nghin cu ton din hn v
hot tnh khng ung th ca cc hp cht cembranoit.
Bng III.1.5.2. Hot tnh gy c t bo ca cc cht
Hp cht
DMSO
Ellipthine
SM1
SM2
SM3
SM4
SM5
SM6
SM7
SM8
SM9
SM10

Gi tr CS% (Cell survival)

IC50 (g/ml)

KB

FL

Hep-G2

KB

FL

Hep-G2

1000,0

1000,0

100,00,0

1,071,07
40,20,5

1,340,63
44,21,5
39,21,1

0,22

0,11

0,19

72,12,6

1,520,00
31,10,7
34,22,5

39,40,7

74,22,2

76,7

3,23
(-)
3,77
(-)
4,23
(-)
4,32
2,23
(-)
(-)

2,77
3,06
(-)
(-)
1,97
4,21
(-)
3,57
(-)
(-)

3,45
3,69
(-)
(-)
3,95
3,55
(-)
2,65
(-)
(-)

72,62,5

84,91,4

66,71,8

44,20,9

34,21,6
29,22,6

46,71,5
24,21,9

21,11,2
43,22,1
69,81,4
41,12,2

65,73,4

78,15,6

66,72,3

79,62,1

74,93,4

56,70,8

56,13,2

74,23,2

32,20,7

219

III.1.5.3. Hot tnh chng xy ha

Kt qu nh gi hot tnh chng xy ha thng qua phn ng bao vy gc
t do DPPH cho thy, ch c hp cht SM4 v SM6 th hin hot tnh vi gi tr
ED50 tng ng l 75,5 v 62,3 g/ml. iu ny cho thy s ph hp v theo ti
liu cng b, (+)-sarcophine (SM6) th hin hot tnh chng xy ha, c
nh gi bng kh nng c ch s sn sinh ion m peoxit (O2-) v thromboxane B2
(TXB2) bi cc t bo tiu thn kinh m no chut s sinh c hot ha
vi lipopolysaccharit ca E. coli (LPS) [181] khi c kch thch bng phorbol
12-myristate 13-acetate (PMA). Hot tnh chng xy ha ca SM4 v SM6 c
th c gii thch bi s c mt ca nhm ktn trong phn t. SM6 c phn
lp vi hm lng kh cao, iu ny gi m cho nhng nghin cu ng dng tip
theo ca hp cht ny theo nh hng hot tnh chng xy ha.
Bng 1.5.3. Hot tnh chng xy ha thng qua kh nng thu dn gc t do
DPPH ca cc hp cht.
Hp cht
Chng (-)
Chng (+)
SM1
SM2
SM3
SM4
SM5
SM6
SM7
SM8
SM9
SM10

Gi tr SC %
(scavenging capacity)
0,000,00
81,220,52

ED50 (g/ml)
>100
20,1
(-)
(-)
(-)
75,5
(-)
62,3
(-)
(-)
(-)
(-)

34,211,81
42,552,11
23,771,44

55,870,72
43,213,65

62,450,52
44,211,78
34,551,75
33,471,28
46,232,01

T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc

loi san h mm Sarcophyton mililatensis, chng ti bin son v gi ng ti
2 cng trnh trn cc tp ch chuyn ngnh quc t.
- Chau Van Minh, Nguyen Xuan Cuong, Tran Anh Tuan, Eun Mi Choi, Young Ho Kim,
and Phan Van Kiem, A new 9,11-secosterol from the Vietnamese Sea soft coral
Sarcophyton mililatensis, increases the function of osteoblastic MC3T3-E1 cells. Natural
Product Communications (USA), 2007, 2(11), 1095-1100.
- Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van Kiem, Chau Van Minh, Eun Mi Choi,
and Young Ho Kim, New Cembranoid Diterpenes from the Vietnamese Soft Coral
Sarcophyton mililatensis Stimulate Osteoblastic Differentiation in MC3T3-E1 Cells.
Chemical and Pharmaceutical Bulletin (Japan), 2008, 56(7), 988-992.

220

Ngoi ra, chng ti cn son tho v ng ti 02 cng trnh trn tp ch

chuyn ngnh quc gia l:
- Chau Van Minh, Nguyen Xuan Cuong, Tran Anh Tuan, Eun Mi Choi, Young Ho Kim,
and Phan Van Kiem, Polyhydroxylated sterols from the Vietnamese Sea soft coral
Sarcophyton mililatensis, increases the function of osteoblastic MC3T3-E1 cells. Tp ch
Khoa hc (i hc S phm H Ni II), 2007, 1(1), 49-54.
- Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van Kiem, and Chau Van Minh,
Sarcophytol X, a new cembranoid diterpene from the vietnamese sea soft coral
Sarcophytol mililatensis. Tp ch Khoa hc (i hc S phm H Ni II), 2007, 1(1), 5560.

Cc kt qu trn cng c s dng trong 01 lun vn thc s ha hc

vi ti: Nghin cu thnh phn ha hc v hot tnh sinh hc loi san h mm
Sarcophyton mililatensis, ThS Trn Anh Tun.

221

Bng tng hp cc cht phn lp c t loi san h mm

Sarcophyton mililatensis
18

4
7

HO

16

21

OMe

19

15

17

OH

14

11
20

SM1: (-)-7-hydroxy-8methoxydeepoxysarcophytoxide (cht mi)

SM2 : (-)-17-Hydroxysarcophytonin A
(cht mi)
O

OH

OH

OH

OH
OH

SM4 : Sarcophytol X (cht mi)

SM3 : Sarcophytol V (cht mi)

H
HO

HO

SM5 : (+)-7,8-dihydroxydeepoxysarcophytoxide (cht mi)

SM6: (+)-Sarcophine
28

21

26
24

18

17

11

HOH2C

27

13

19

25

20

14

1
8

10
3

5
6

HO

SM7: Sarcophytoxide

OH

SM8: Sarcomilasterol (cht mi)

28

28

21

21

26

26

24
18

HO
19

20
17

13

11

24
25

18

17

27

OH
19

13

11

25

27

10
3

20

10
7

HO

HO

OH

OH

OH

OH

SM9: Sarcoaldesterol B

SM10: Ergosta-1,3,5,6-tetraol.

222

III.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca

loi san h mm cladiella sp. (ALcyoniidae)
III.2.1. M u
Mu san h mm Cladiella sp. c thu thp ti Cn C, tnh Qung Tr
vo thng 11 nm 2006 v thng 3 nm 2007 v c TS. Cng Thung v cc
cng s, Vin Ti nguyn v Mi trng Bin, Vin KH&CN Vit Nam gim
nh tn khoa hc.

- c im: Tp on
thng c thn ngn, b mt
tp on cng. Polyparium
c nhiu cc thu ngn ,
thu c dng ngn hoc u
bu. Polyp n. Gai xng
c 2 u chiu di thng
<0,15mm
- Mu sc: c mu hi
trng, nu nht, hoc xanh
nht

San h mm: Cladiella sp

Cladiella l mt chi san h mm sinh sng kh ph bin ti cc vng bin
Vit Nam. Hin nay nc ta hon ton cha c cng trnh khoa hc no c
cng b v thnh phn ha hc v hot tnh sinh hc ca cc loi thuc chi ny.
Trn th gii hin cng cha c nhiu cng trnh c cng b v chi ny v mi
c khong 36 hp cht c phn lp v xc nh cu trc. Cc lp cht chnh ca
chi ny l ceramide, cerebroside, steroit v ditecpenoit.
Di y, chng ti gii thiu mt s hp cht i din c phn lp
t chi Cladiella.
OH

OH
O

1'

20'

4"

6"

HO
HO

1'

15

HN

OH
O

5"
2"

OH

3"

19'

HN

O
18
1

1"

HO

HO

18

19

HO

n = 16, 17, 18 [196]

Humesamide [197]
OH

OH
O
O

O
O

HO

OH

Epoxycladine B [198]

Cladiellisin [199]

223

O
O

OH

OH

Cladiellaperoxide [199]
OH

Acetoxycladiellin [200]

OH

OH
OH

OH
O

OH

Sclerophytin C [201]

Sclerophytin E [202]

HO
O
OH
HO

HO

OH

Cladiellin A [203]
[204]
HO

HO

OH

O
OH

HO

OH

16

[206]

[205]

III.2.2. Phn lp cc hp cht

Mu san h mm ti (2 kg) c xay nh, sau chit vi methanol ba
ln, mi ln 72 gi. Cc dch chit methanol c gp li v ct loi dung mi
di p sut gim thu c cn chit MeOH th. Cn chit ny c ha vo
nc ri tin hnh loi mui trn ct Dianion vi dung mi ra gii l nc.
Dng MeOH ra gii thu hi cht thu c 20 gam dch c. Dch c ny c
ho tan vo 01 lt nc ct sau c chit ln lt bng n-hexan, cloroform, v
BuOH (mi ln 02 lit) thu c cc cn dch tng ng: hexan (5 g), cloroform (7
g), BuOH (4 g) v nc (4 g) (S III.2.2.a).
Cn chit n-hexan: SH (5 g) c tin hnh phn tch trn sc k ct lp
li nhiu ln vi cc h dung mi thch hp (s III.2.2.b) thu c cc hp
cht SH-7 v SH-8.
224

San h mm Cladiella sp. ti

(2 kg)
- Chit MeOH, ct loi dung mi di p sut
gim
- Loi mui bng cch chy qua ct Dianion vi
dung mi ra gii l nc ct, sau thu hi
cht bng MeOH.
B sung nc

SM - Cn chit MeOH (20g)

B sung n-hexan
n-Hexan: nc 1/1

Lp nc
B sung CHCl3

SH - Cn n-hexan (5 g)

CHCl3: nc 1/1

Lp nc
B sung BuOH

SC - Cn CHCl3 (7 g)

BuOH: nc 1/1
lp nc

SB - Cn BuOH (4 g)

SN - Cn nc (4 g)

S III.2.2.a. S chit phn on mu san h mm

SH - (5 g)
Silica gel CC
Hexan/Axeton 100/0-0/100

SH1 (3000mg)

SH2 (400mg)

SH3 (250mg)

Silica gel CC
n-hexan/Etylaxetat
10/1
SH-8
(200mg)

SH4 (750mg)

SH5 (600g)

Silica gel CC
n-hexan/axeton
7/1
SH-7
(100mg)

S III.2.2.b. S phn lp cc hp cht t cn chit n-hexan

225

Cn chit clorofrom: SC (7 g) c tin hnh chy ct th trn sc k ct

pha thng gradient nng metanol trong cloroform thu c 5 phn on nh
hn l SC1-5. Cc phn on SC1, SC4 v SC5 c tip tc phn tch trn sc k
ct pha thng vi h dung mi ra gii thch hp thu c cc hp cht SH-3, 4,
5, v 6 nh c ch ra trn s III.2.2.c. Bng cc phng php tng t (s
III.2.2.d), cc hp cht SH-1 v SH-2 c phn lp t cn chit BuOH.
SC (7 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

SC1 (2g)

SC2 (1,2g)

SC3 (1,3g)

SC5 (1g)

Silica gel CC
n-hexan/axeton
3/2

Silica gel CC
Cloroform/axeton: 10/1

SH-5
(500mg)

SC4 (1,5g)

SH-6
(70mg)

SH-4
(8mg)

Silica gel CC
CHCl3/MeOH
7/1
SH-3
(9mg)

S III.2.2.c. S phn lp cc hp cht t cn chit cloroform

SB - (4 g)
Silica gel CC
Cloroform/MeOH/H2O: 5/1/0 - 2/1/0.2

SB1 (1.0 g)

SB2 (2.5 g)

Silica gel CC
CHCl3/MeOH
7/1
SH-1
(4mg)

SB3 (1.5 g)

SB4 (600mg)

SB5 (1.4 g)

Silica gel CC
Etylaxetat/CHCl3/MeOH
6/2/1
SH-2
(12mg)

S III.2.2.d. S phn lp cc hp cht t cn chit butanol

226

III.2.3. Hng s vt l v d kin ph ca cc hp cht

III.2.3.1. Hp cht SH1: Cladiasterol (Cht mi: 3,11,16-trihydroxy-24methylene-9,11-secocholest-5-en-9-one-3-O--L-fucopyranoside)
21
Cht bt v nh hnh mu trng; []D -115o (c 0.50, CHCl3); positive ESI-MS:
m/z 615.2 [M+Na]+, 454.2 [M-fucose+Na]+; negative ESI-MS: m/z 591.4 [M-H](C34H56O8).
1
H-NMR (500 MHz, CDCl3) (ppm): 1,53 (1H, m, Ha-1), 1,87 (1H, m, Hb-1),
1,65 (1H, m, Ha-2), 1,99 (1H, m, Hb-2), 3,49 (1H, m, H-3), 2,27 (1H, m, Ha-4), 2,50
(1H, m, Hb-4), 5,53 (1H, d, J = 5,5 Hz, H-6), 2,05 (1H, m, Ha-7), 2,53 (1H, m, Hb-7),
3,19 (1H, m, H-8), 3,78 (1H, m, Ha-11), 3,83 (1H, m, Hb-11), 1,40 (1H, m, Ha-12), 1,80
(1H, m, Hb-12), 2,65 (1H, m, H-14), 1,40 (1H, m, Ha-15), 2,12 (1H, m, Hb-15), 4,37
(1H, m, H-16), 1,49 (1H, m, H-17), 0,90 (3H, s, H-18), 1,40 (3H, s, H-19), 1,86 (1H, m,
H-20), 1,09 (3H, d, J = 6,5 Hz, H-21), 1,33 (1H, m, Ha-22), 1,72 (1H, m, Hb-22), 1,97
(1H, m, Ha-23), 2,19 (1H, m, Hb-23), 2,27 (1H, m, H-25), 1,05 (3H, s, H-26), 1,05 (3H,
s, H-27), 4,72 (1H, d, J = 1,0 Hz, Ha-28), 4,78 (1H, s, Hb-28), 5,02 (1H, s, H-1), 3,82
(1H, m, H-2), 3,77 (1H, m, H-3), 3,82 (1H, m, H-4), 4,05 (1H, dd, J = 6,5, 13,0 Hz)
v 1,30 (3H, s, H-6).
13
C-NMR (125 MHz, CDCl3) (ppm): 31,15 (t, C-1), 28,74 (t, C-2), 77,10 (d, C3), 37,26 (t, C-4), 139,64 (s, C-5), 121,91 (d, C-6), 32,83 (t, C-7), 43,45 (d, C-8), 217,30
(s, C-9), 48,52 (s, C-10), 59,41 (t, C-11), 41,81 (t, C-12), 45,24 (s, C-13), 40,44 (d, C14), 35,68 (t, C-15), 72,04 (d, C-16), 54,28 (d, C-17), 18,53 (q, C-18), 22,88 (q, C-19),
29,75 (d, C-20), 17,55 (q, C-21), 34,95 (t, C-22), 31,25 (t, C-23), 156,83 (s, C-24), 34,03
(d, C-25), 21,84 (q, C-26), 21,97 (q, C-27), 106,46 (t, C-28), 97,11 (d, C-1), 69,37 (d,
C-2), 71,69 (d, C-3), 71,66 (d, C-4), 66,15 (d, C-5) v 16,18 (q, C-6).

III.2.3.2. Hp cht SH2: Cladiaceramide (Cht mi: (2S,3R,4E,8E)-[(2R)-2hydroxy-hexadecanoylamino]-4,8-octadecadiene-1,3-diol)

28
Cht bt v nh hnh mu trng; []D -11.1o (c=0.5, CHCl3); positive ESI-MS:
m/z 574.4 [M+Na]+, 552.4 [M+H]+, 534.4 [M-H2O+H]+ 282.1; negative ESI-MS: m/z
550.4 [M-H]-, 297.0 (C34H65NO4, M=551).
1
H-NMR (500 MHz, CDCl3) (ppm): 3,69 (1H, dd, J = 3,5, 11,5 Hz, H-1a), 3,80
(1H, dd, J = 5,0, 11,5 Hz, H-1b), 3,86 (1H, m, H-2), 4,17 (1H, t, J = 5,5 Hz, H-3), 5,49
(1H, dd, J = 6,5, 15,5 Hz, H-4), 5,76 (1H, dt, J = 6,5, 15,5 Hz, H-5), 2,09 (2H, m, H-6),
2,09 (2H, m, H-7), 5,40 (1H, dd, J = 6,5, 15,5 Hz, H-8), 5,40 (1H, dd, J = 6,5, 15,5 Hz,
H-9), 1,96 (2H, m, H-10), 1,26 (36H, br s, H-11 to H-17 and H-5 to H-15), 0,88 (6H, t,
J = 6,5 Hz, H-18 and H-16), 4,04 (1H, dd, J = 3,5, 8,0 Hz, H-2), 1,78 (1H, m, Ha-3),
1,57 (1H, m, Hb-3), 1,41 (2H, m, H-4) v 7,34 (1H, d, J = 8,5 Hz, NH).
13
C-NMR (125 MHz, CDCl3) (ppm): 61,57 (t, C-1), 54,61 (d, C-2), 73,24 (d, C3), 129,33 (d, C-4), 133,66 (d, C-5), 32,65 (t, C-6), 32,36 (t, C-7), 131,33 (d, C-8),
129,33 (d, C-9), 32,77 (t, C-10), 29,40-29,89 (t, C-11 to C-15), 32,11 (t, C-16), 22,85 (t,
C-17), 14,2 (q, C-18), 176,17 (s, C-1), 72,24 (d, C-2), 34,68 (t, C-3), 25,38 (t, C-4),
29,40-29,89 (t, C-5 to C-13), 32,11 (t, C-14), 22,85 (t, C-15) v 14,2 (q, C-16).

227

III.2.2.3. Hp cht SH3: Cladiacerebroside (Cht mi: 1-O-(2-sulfo--Dglucopyranosyl)-(2S,3R,4E,8E)-2-[(2R)-2-hydroxyeicosanoylamino]-9-methyl4,8-octadecadiene-1,3-diol).

Cht bt v nh hnh mu trng, postitive ESI-MS m/z: 848,6 [M+H]+, negative
ESI-MS m/z: 846,8 [M-H]- (C45H85NO11S, M = 847).
1
H-NMR (500 MHz, CDCl3) (ppm): 3,76 (1H, Ha-1), 4,03 (1H, Hb-1), 3,84 (1H,
dd, J = 2,5, 12,0 Hz, H-2), 4,11 (1H, t, J = 5,5 Hz, H-3), 5,46 (1H, dd, J = 6,5, 15,5 Hz,
H-4), 5,74 (1H, dd, J = 6,5, 15,5 Hz, H-5), 2,05 (4H, m, H-6 v H-7), 5,10 (1H, H-8),
1,95 (2H, m, H-10), 1,26 (46H, br s, H-11 n H-17 v H-5 n H-19), 0,88 (6H, t, J
= 6,5 Hz, H-18 v H-20), 1,57 (3H, s, H-19), 4,03 (1H, H-2), 1,76 (1H, Ha-3), 1,54
(1H, Hb-3), 1,38 (2H, m, H-4), 4,27 (1H, d, J = 7,5 Hz, H-1), 3,25 (1H, m, H-2),
3,42 (1H, m, H-3), 3,44 (1H, m, H-4), 3,28 (1H, m, H-5), 3,75 (1H, m, Ha-6) v
4,01 (1H, m, Hb-6).
13
C-NMR (125 MHz, CDCl3) (ppm): 68,54 (t, C-1), 53,29 (d, C-2), 72,29 (d, C3), 128,76 (d, C-4), 134,21 (d, C-5), 32,69 (t, C-6), 28,13 (t, C-7), 123,27 (d, C-8),
136,19 (s, C-9), 39,79 (t, C-10), 29,43-29,79 (t, C-11 n C-15 v C-5 n C-17),
31,98 (t, C-16 v C-18), 22,75 (t, C-17 v C-19), 14,13 (q, C-18 v C-20), 15,96 (q, C19), 176,05 (s, C-1), 73,44 (d, C-2), 34,57 (t, C-3), 25,33 (t, C-4), 103,10 (d, C-1),
76,15 (d, C-2), 76,32 (d, C-3), 69,71 (d, C-4),77,44 (d, C-5) v 61,35 (t, C-6).

III.2.3.4. Hp cht SH4: 3,6,11-trihydroxy-24-methylene-9,11-secocholest-7en-9-one

Tinh th hnh kim mu trng, nhit nng chy: mp. 162-164oC; quay
20
quang hc [ ]D +9o (c 0,24, CHCl3); positive ESI-MS: m/z 447,2 [M + H]+, negative ESIMS: m/z 445,4 [M - H]- (C28H46O4).
1
H-NMR (500 MHz, CDCl3) (ppm): 1,46 (Ha-1), 1,90 (Hb-1), 1,45 (Ha-2), 1,92
(Hb-2), 3,58 (m, H-3) 1,42 (Ha-4), 2,30 (Hb-4), 1,76 (H-5), 4,26 (d, J = 8,0 Hz, H-6),
6,56 (d, J = 1,5 Hz, H-7), 3,65 (m, Ha-11), 3,85 (m, Hb-11), 1,12 (Ha-12), 1,62 (Hb-12),
3,42 (t, J = 10,5 Hz, H-14), 1,60 (H-15), 1,45 (Ha-16), 1,88 (Hb-16), 1,75 (H-17), 0,66
(s, H-18), 1,13 (s, H-19), 1,45 (H-20), 0,99 (d, J = 6,5 Hz, H-21), 1,17 (Ha-22), 1,58
(Hb-22), 1,92 (Ha-23), 2,12 (Hb-23), 2,23 (H-25), 1,03 (d, J = 6,5 Hz, H-26), 1,03 (d, J =
6,5 Hz, H-27), 4,66 (s, Ha-28) v 4,73 (s, Hb-28).
13
C-NMR (125 MHz, CDCl3) (ppm): 32,07 (t, C-1), 30,71 (t, C-2), 70,12 (d, C3), 33,00 (t, C-4), 48,86 (d, C-5), 69,54 (d, C-6), 147,16 (d, C-7), 137,33 (s, C-8), 205,07
(s, C-9), 45,00 (s, C-10), 59,45 (t, C-11), 41,35 (t, C-12), 46,39 (s, C-13), 42,85 (d, C14), 26,88 (t, C-15), 26,28 (t, C-16), 50,09 (d, C-17), 17,49 (q, C-18), 16,21 (q, C-19),
34,82 (d, C-20), 18,99 (q, C-21), 34,32 (t, C-22), 31,84 (t, C-23), 156,70 (s, C-24), 33,98
(d, C-25), 21,92 (q, C-26), 22,05 (q, C-27) v 106,34 (t, C-28).

III.2.3.5. Hp cht SH5: Cholesterol

Cht kt tinh mu trng, nhit nng chy: 148-149 oC, C27H46O, M=386.
1
H-NMR (500 MHz, CDCl3) (ppm): 3,52 (1H, m, H-3), 5,35 (1H, br d, J = 5,0
Hz, H-6), 1,00 (3H, s, H-19), 0,91 (3H, s, H-21), 0,86 (6H, d, J = 6.5 Hz), H-26, 27) v
0.68 (3H, s, H-18).

228

C-NMR (125 MHz, CDCl3) (ppm): 37.2 (t, C-1), 31.6 (t, C-2), 71.8 (d, C-3),
42,2 (t, C-4), 140,7 (s, C-5), 121,7 (d, C-6), 31,8 (t, C-7), 31,8 (d, C-8), 50,1 (d, C-9),
36,5 (s, C-10), 21,1 (t, C-11), 39,8 (t, C-12), 42,3 (s, C-13), 56,7 (d, C-14), 24,3 (t, C15), 28,3 (t, C-16), 56,1 (d, C-17), 11,9 (q, C-18), 19,4 (q, C-19), 35,7 (d, C-20), 18,7 (q,
C-21), 36,1 (t, C-22), 23,8 (t, C-23), 39,5 (t, C-24), 28,0 (d, C-25), 22,5 (q, C-26) v
22,8 (q, C-27).
13

III.2.3.6. Hp cht SH6: Chimyl alcohol

Tinh th mu trng, nhit nng chy: mp. 64-65oC; ph khi lng ESI-MS
m/z: 317,1 [M+H]+ (positive), (C19H40O3, M=316).
1
H-NMR (500 MHz, CDCl3) (ppm): 3,64 (1H, dd, J = 3,0 and 11,0 Hz, H-1a),
3,71 (1H, dd, J = 5,0 and 11,0 Hz, H-1b), 3,86 (1H, m, H-2), 3,52 (2H, m, H-3), 3,45
(2H, m, H-1), 1,58 (2H, m, H-2), 1,26 (2H, br s, H-3 to H-17) v 0,88 (3H, t, J = 7,0
Hz, H-18) ppm.
13
C-NMR (125 MHz, CDCl3) (ppm): 72,5 (t, C-1), 70,4 (d, C-2), 64,3 (t, C-3),
71,9 (t, C-1), 2,9,3-29,7 (t, C-2), 26,1 (t, C-3), 29,3 - 29,7 (t, C-4 to C-13), 31,9 (t,
C-14), 22,7 (t, C-15) v 14,2 (q, C-16) ppm.

III.2.3.7. Hp cht SH7: Batyl alcohol

Tinh th mu trng; nhit nng chy: mp. 70-71oC; ph khi lng ESI-MS
m/z: 345,1 [M+H]+ (positive), (C21H44O3, M=344).
1
H-NMR (500 MHz, CDCl3) H(ppm): 3,62 (1H, dd, J = 3,0 and 11,0 Hz, Ha-1),
3,71 (1H, dd, J = 5,0 and 11,0 Hz, Hb-1), 3,86 (1H, m, H-2), 3,52 (2H, m, H-3), 3,45
(2H, m, H-1), 1,57 (2H, m, H-2), 1,26 (2H, br s, H-3 to H-17) v 0,88 (3H, t, J = 7,0
Hz, H-18) ppm.

III.2.3.8. Hp cht SH8: 1-heptadecanol

Tinh th mu trng; nhit nng chy 59,5-60.0oC; ph khi lng postitive
ESI-MS m/z 279.0 [M+Na]+, negative ESI-MS m/z 255.0 [M-H]-, 237.0 [M-H2O-H]-,
(C17H36O, M=256).
1
H-NMR (500 MHz, CDCl3) H(ppm): 3,63 (2H, t, J = 7,0 Hz, H-1), 1,57 (2H,
m, H-2), 1,26 (30H, br s, H-3 to H-17) v 0,88 (3H, t, J = 7,0 Hz, H-18) ppm.

229

III.2.4. xc nh cu trc ha hc ca cc hp cht

III.2.4.1. Hp cht SH1: Cladiaserol (Cht mi: 3,11,16-trihydroxy-24methylene-9,11-secocholest-5-en-9-one-3-O--L-fucopyranoside)
Hp cht SH1 nhn c di dng bt v nh hnh mu trng. Cng
thc phn t ca n c xc nh l C34H56O8 (tng ng vi khng no l 7)
bng cc d kin ph khi lng phn gii cao (HR-ESI-MS): ti m/z 614,389
(tnh ton l thuyt cho C34H56O8Na: 615,3873) v ph khi lng (positive ESIMS ti m/z 615,2 [M+Na]+, 454,2 [M-fucose+Na]+; negative ESI-MS: m/z 591,4
[M-H]-) v ph cng hng t ht nhn. Pc ion phn mnh ti m/z 454.2 trn ph
khi lng th hin s phn ct mt ng 6 cacbon ca SH1.

Hnh III.2.4.1.a. Ph 1H-NMR ca SH1

Cc ph NMR ca SH1 cho thy y l mt hp cht 9,11-secosterol
(Bng III.2.4.1). Trn ph 1H- v 13C-NMR xut hin cc tnh hiu c trng ca
mt n v ng -L-fucose ti 16,2 (q)/1,30 (3H, d, J = 6,5Hz), 66,2 (d)/4,05
(1H, dd, J = 6,5 and 13,0 Hz), 69,4 (d)/3,82 (1H, m), 71,6 (d)/3,82 (1H, m), 71,7
(d)/3,77 (1H, m) v 97,1 (d)/ 5,02 (1H, s) [207-208]. Trn ph 13C-NMR cc tn
hiu ti 217,3 (s), 121,9 (d)/139,6 (s) v 106,5 (t)/156,8 (s) c trng tng ng
cho mt nhm ketone, mt ni i b th 3 v tr v mt ni i cui mch (mt
u t do). Ngoi ra s xut hin ca mt nhm oximetylen, 2 nhm oxymetin v
5 nhm metyls cng c xc nh tng ng bi cc tn hiu cng hng ti
59,4 (t), 72,0 (d), 77,1 (d), 17,5 (q), 18,5 (q), 21,8 (q), 22,0 (q) v 22,9 (q).

230

28
26

21

HO

20

11

22

23

12
19
6'
5'

HO

3'

HO

2'

10
5

3
1'

4'

13

25

27

17
16

14

1
2

24

18

15

OH

OH

Hnh III.2.4.1.b: Cu trc ha hc ca SH1

Hnh III.2.4.1.c. Ph 13C-NMR ca SH1

Bng III.2.4.1: Kt qu ph cng hng t ht nhn ca SH1
TT

Ca, b

1
2
3
4
5
6

31,1
28,7
77,1
37,3
139,6
121,9

32,8

8
9
10

43,4
217,3
48,5

Ha, c
(mult., J in Hz)
1,53 (m)/1,87 (m)
1,65 (m)/1,99 (m)
3,49 (m)
2,27 (m)/2,50 (m)
5,53 (d, 5,5)
2,05 (m)
2,53 (m)
3,19 (m)
-

231

HMBC
(H to C)

C-4, 7, 8, 10

C-7, 9, 13, 14

11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27

59,4
41,8
45,2
40,4
35,7
72,0
54,3
18,5
22,9
29,7
17,5
34,9
31,2
156,8
34,0
21,8
22,0

28

106,4

1
2
3
4
5
6

97,1
69,4
71,7
71,6
66,1
16,2

3,78 (m)/3,83 (m)

1,40 (m)/1,80 (m)
2,65 (m)
1,40 (m)/2,12 (m)
4,37 (m)
1,49 (m)
0,90 (s)
1,40 (s)
1,86 (m)
1,09 (d, 6,5)
1,33 (m)/1,72 (m)
1,97 (m)/2,19 (m)
2,27 (m)
1,05 (d, 6,5)
1,05 (d, 6,5)
4,72 (d, 1,0)
4,78 (s)
5,02 (s)
3,82 (m)
3,77 (m)
3,82 (m)
4,05 (dd, 6,5, 13,0)
1,30 (d, 6,5)

C-11, 13, 14, 17

C-8, 9, 13, 15, 18
C-13, 14, 17

C-13, 14, 17
C-1, 5, 9, 10
C-17, 20, 22
C-28

C-24, 27
C-24, 26
C-23, 24, 25
C-3, 5

C-5

o trong CDCl3, b 125 MHz, c 500 MHz

Hnh III.2.4.1.d. Ph 13C-NMR v cc ph DEPT ca SH1

232

 xc nh cu trc tng th ca SH1, cc ph 2D-NMR c o. Ph

HSQC cho php xc nh cc proton gn trc tip vi cacbon tng ng (Bng
III.2.4.1). Xem xt mc khng no cng vi cc d kin nu cho php khng
nh phn aglycon ca SH1 phi c cu trc 3 vng v do n l mt
secosterol. C th l cu trc 9,11-secosterol vi nhm hydroxy ti C-11 c
khng nh bi s xut hin cc tnh hiu cng hng c trng ca mt h
proton tng tc c lp bao gm cc cp proton trn cng mt cacbon tng tc
vi nhau cng hng ti 1,40 (H-12a), 1,80 (H-12b), 3,78 (H-11a), v 3,83 (H11b), c gn vi 2 cacbon cng hng ti 59,4 (C-11) v 41,8 (C-12) thng
qua ph HSQC. C s liu ph 1H- v 13C-NMR ca mnh etyl ny u hon ton
ph hp vi cc gi tr tng ng ca cc hp cht 9,11-secosterol c cu trc
tng t [209-212]. Ngoi ra, s c trng cho cu trc 9,11-secosterol cng th
hin s xut hin ca nhm ketone ti C-9 ( 217,3), m v tr ca n c xc
nh bi cc tng tc xa vi cc proton ca nhm C-19 metyl ti 1,40 v vi
cc proton metin ti 3,19 (H-8) v 2,65 (H-14).
S chuyn dch v vng trng thp ti v tr C-3 ( 77,1) gi rng phn
t ng gn vo C-3 v iu ny c khng nh bi cc tng tc xa ca H-1
vi C-3 v ca H-3 vi C-1 trn ph HMBC. Cu trc 5, 11-hydroxy v cu trc
nhnh bn c xc nh bng cch so snh cc gi tr ph 1H- v 13C-NMR ti
cc v tr tng ng vi cc ti liu cng b ca cc hp cht c cu trc tng
t [208, 211, 212] v c khng nh li bng cc tng tc trn ph HMBC.

Hnh III.2.4.1.e. Ph HSQC ca SH1

233

Phn tch chi tit cc tng tc trn ph 1H-1H COSY cho php ghp ni
cu trc t H-1 n H-4; H-6 n H-24; v H-11 n H-12. Trong , H-8 (
3,19) tng tc vi H-14 ( 2,65), proton ny li th hin tng tc vi H-15 (
1,40 v 2,12). Proton H-15 li tng tc tip vi H-16 ( 4.37). Cc tng tc
gia H-16 vi H-17 ( 1,49) v H-17 vi H-18 ( 1,86) cng xut hin trn ph
COSY. Cc d kin trn, cng vi cc tng tc trn ph HMBC (Bng III.2.4.1,
Hnh III.2.4.1.g) cho php khng nh v tr ca nhm C-16 hydroxyl cng nh
ton b cu trc phng ca SH1.

Hnh III.2.4.1.f. Ph HMBC ca SH1

HO

OH
H

HO

HMBC
COSY

HO

OH

Hnh III.2.4.1.g. Mt s tng tc COSY v HMBC chnh ca SH1

234

Cu hnh ti cc tm bt i ca SH1 c xc nh bng ph ROESY,

hng s tng tc v s so snh s liu ph NMR ca n vi cc hp cht
sercosterol c cu trc tng t. Theo cc ti liu cng b, tn hiu cng
hng ti 3,49 (m) c trng cho proton H-3 vi cu hnh [207] v tn hiu
ca proton H-8 ti 3,19 (m) c trng cho cu hnh (ti 2,6 n 2,7 cho cu
hnh ) [211].

Hnh III.2.4.1.h. Ph COSY ca SH1

Tng tc ROESY gia H-8 ( 3,19, m) vi H3-18 ( 0,90, s) v H3-19 (
1,40, s) (Hnh III.2.4.1.j) cho thy chng u c cu hnh . Vi cu trc lp th
ti cc v tr nu trn c xc nh, ph ROESY s l mt cng c hu hiu
xc nh cu hnh ti cc tm bt i cn li. S xut hin tn hiu tng tc
gia proton metin C-17 ( 1,49, m) vi cc proton metin C-14 ( 2,65, m) v C-16
cho thy chng c cng dng cu hnh do cu hnh ca nhm hydroxyl C-16
phi l .

235

Hnh III.2.4.1.i. Ph HMBC ca SH1

CH3

O
O
H3C

CH3

HO

CH3
OH
H

H
H

OH

HO
OH

Hnh III.2.4.1.j. Mt s tng ROESY quan trng ca SH1

T tt c cc phn tch nu phn trn, cu trc ha hc ca SH1 c
xc nh l 3,11,16-trihydroxy-24-methylene-9,11-secocholest-5-en-9-one-3O--L-fucopyranoside. y l mt hp cht mi v c chng ti t tn l
Cladiasterol. Mt iu th v hn na, y l mt hp cht 9,11-secosterol
glycoside u tin c cng b cho ti nay.

236

Hnh III.2.4.1.k. Ph ESI-MS ca SH1

cald for [C34H56O8Na]+ 615.3873

Hnh III.2.4.1.l. Ph HR-ESI-MS ca SH1

III.2.4.2. Hp cht SH2: (2S,3R,4E,8E)-[(2R)-2-hydroxyhexadecanoylamino]-4,8-octadecadiene-1,3-diol (cht mi)
Hp cht SH2 nhn c di dng cht bt mu trng. Trn ph khi
lng phn gii cao (HR-ESI-MS) xut hin pic ion phn t ti m/z 574,489 (tnh
ton l thuyt cho C34H65NO4Na: 574,4811) v trn ph positive ESI-MS xut

237

hin pc ion phn t ti m/z 574.4 [M+Na]+ v 552.4 [M+H]+. Cng vi cc ph

NMR cho php xc nh cng thc phn t ca SH2 l C34H65NO4 (M=551).

Hnh III.2.4.2.a. Ph 1H-NMR ca SH2

Trn ph 1H-NMR ca SH2 xut hin cc tn hiu c trng cho 2 nhm
metyl ti 0,88 (6H, t, J = 6.5 Hz); cc proton metilen ca mch di ti 1,26
(36H, br s); mt nhm metilen gn vi nguyn t xy ti 3,69 (1H, dd, J = 3,5
v 11,5 Hz) v 3,80 (1H, dd, J = 5.0 v 11.5 Hz); 2 nhm metin gn vi nguyn
t xy ti 3,69 (1H, dd, J = 3,5 v 8,0 Hz) v 4,17 (1H, t, J = 5,5 Hz); 4 proton
olefin ti 5,40 (2H, dd, J = 6,5 v 15,5 Hz), 5,49 (1H, dd, J = 6,5 v 15,5 Hz)
v 5,76 (1H, dd, J = 6,5 v 15,5 Hz) v mt proton amide ti 7,34 (1H, d, J =
8,5 Hz). Trn ph 13C-NMR xut hin cc tn hiu c trng ca mt nhm amide
carbonyl ti 176,2 (s); mt metin gn vi nguyn t nit ti 54,6 (d); mt
metilen gn vi nguyn t xy ti 61,6 (t); 2 metin gn vi nguyn t xy ti
72,2 (d) v 73,2 (d); 2 lin kt i b th 2 v tr ti 129,3 (2, d), 131,3 (d) v
133,7 (d); v 2 nhm metyl ti 14,2 (2, q). Cc d kin ph ny gi rng hp
cht SH2 l mt ceramide [213].
OH
O

1'

3'

10

16'

HN
HO

18
1

OH

Hnh: III.2.4.2.b. Cu trc ca hp cht SH2

238

Tt c cc v tr ca SH2 c gn bng cc phng php ph NMR hai

chiu hin i l 1H-1H COSY, HMQC v HMBC (Hnh III.2.4.2.b).

Hnh III.2.4.2.c. Ph 13C-NMR ca SH2

Bng III.2.4.2: Cc kt qu ph NMR ca SH2
C

[214]

a, b

62,1

61,57

2
3
4
5
6
7
8
9
10
11-15
16
17
18
1
2
3

54,6
72,5
129,3
133,8
32,4
32,1
131,4
129,0
32,6
22,7-31,9

54,61
73,24
129,33
133,66
32,65
32,36
131,33
129,33
32,77
29,40-29,89
32,11
22,85
14,2
176,17
72,24
34,68

4
5-13
14
15
16
NH

14,1
175,2
74,2
34,8

22,7-31,9

14,1
-

25,38
29,40-29,89
32,11
22,85
14,2
-

a, c
(mult. J in Hz)
3,69 (dd, 3,5, 11,5)
3,80 (dd, 5,0, 11,5)
3,86 (m)
4,17 (t, 5,5)
5,49 (dd, 6,5, 15,5)
5,76 (dt, 6,5, 15,5)
2,09 (m)
2,09 (m)
5,40 (dd, 6,5, 15,5)
5,40 (dd, 6,5, 15,5)
1,96 (m)
1,26 (br s)
1,26 (br s)
1,26 (br s)
0,88 (t, 6,5)
4,04 (dd, 3,5, 8,0)
1,78 (m)
1,57 (m)
1,41 (m)
1,26 (br s)
1,26 (br s)
1,26 (br s)
0,88 (t, 6,5)
7,34 (d, 8,5)

o trong CDCl3+MeOD, b 125 MHz, c500 MHz.

239

HMBC
(H to C)
C-2, 3
C-1
C-1, 2, 4, 5
C-2, 3, 6
C-3, 7
C-4, 5, 7, 8
C-6, 6, 8, 9

C-8, 9

C-17
C-1
C-1, 2
C-2

C-15

Hnh III.2.4.2.d. Ph HSQC ca SH2

Hnh III.2.4.2.e. Ph 1H-1H COSY ca SH2

240

Hnh III.2.4.2.f. Ph HMBC ca SH2

Phn tch chi tit cc tng tc trn ph 1H-1H COSY cho php ghp ni
cu trc t H-1 n H-6; H-7 sang H-8; H-9 sang H-10; H-2 sang proton amide;
v H-2 n H-4 (Hnh III.2.4.2.g). T cc phn tch nu trn, cng vi cc tng
tc xa trn ph HMBC ca H-1 sang C-2 v 3; H-3 sang C-1, 2, 4 v 5; H-6 sang
C-4, 5, 7 v 8; H-7 sang C-5, 6, 8 v 9; H-2 sang C-1; H-3 sang C-1 v 2; v
H-4 sang C-2 khng nh v tr ca cc nhm hydroxyl ti C-1, 2 v 3 cng
vi cu trc 4,8 ca SH2.
OH
O

COSY

3'

10

1'

16'

HMBC

HN
HO

18
1

OH

Hnh III.2.4.2.g. Mt s tng tc HMBC chnh ca SH2

Cu trc lp th ti v tr C-1, 3 v 2 c xc nh l 2S, 3R v 2R trn
c s s liu ph NMR v quay quang hc ca SH2 hon ton ph hp vi s
liu tng ng ca (2S,3R,4E,8E)-[(2R)-2-hydroxyheptadecanoylamino]-4,8octadeca-diene-1,3-diol [214]. Cu trc hnh hc ca cc lin kt i c xc
nh l u dng trans (4E,8E) trn c s hng s tng tc ln (J = 15,5 Hz)
ca cc proton. Theo cc ti liu cng b, cc ceramide c phn lp cho
ti nay nu nhnh baz mch di c cha 2 ni di ti C-4 v C-8 u c di
mch l 18 cacbon. iu ny hon ton ph hp v cng t chi Cladiella, hp
cht Humesamide c phn lp v cng c cha nhnh baz mch di ny
241

[197]. T tt c cc phn tch nu trn, cng vi s liu ph khi lng ESI-MS

cho php khng nh cu trc ca SH2 l (2S,3R,4E,8E)-[(2R)-2hydroxyhexadecanoylamino]-4,8-octadecadiene-1,3-diol. y l mt hp cht
mi v c chng ti t tn l cladiaceramide.

Hnh III.2.4.2.h. Ph ESI-MS ca SH2

cald for [C34H65NO4Na]+: 574.4811

Hnh III.2.4.2.i. Ph HR-ESI-MS ca SH2

242

III.2.4.3. Hp cht SH3: (Cht mi: 1-O-(2-sulfo--D-glucopyranosyl)(2S,3R,4E,8E)-2-[(2R)-2-hydroxyeicosanoylamino]-9-methyl-4,8octadecadiene-1,3-diol)

Hp cht SH3 cng nhn c di dng bt v nh hnh mu trng.
Cng thc phn t ca n c xc nh l C45H85NO11S (M = 847) bng cc d
kin ph khi [m/z 848.6 [M+H]+ (postitive), 846.8 [M-H]- (negative)] v ph
cng hng t ht nhn.

Hnh III.2.4.3.a. Ph 1H-NMR ca SH3

OH
O

1'

16

4"

6"

HO
HO

HN

OH
O
5"
2"

3"

20'

1"

18
1

SO3H
HO

19

Hnh III.2.4.3.b. Cu trc ha hc ca SH3

Ph NMR ca SH3 tng t nh ca SH2, ngoi tr s xut hin thm cc
tn hiu ca mt n v ng glucose v mt nhm metyl. Trn ph 1H-NMR
xut hin cc tn hiu c trng ca 3 nhm metyl ti H 0,88 (6H, t, J = 7.0 Hz)
v 1,47 (3H, s), tn hiu b che lp ca hai mch hydrocacbon no di ti H 1,26
(46H, br s), mt proton gn vi cacbon anome ti H 4,27 (1H, d, J = 7,5 Hz).

243

Hnh III.2.4.3.c. Ph 13C-NMR ca SH3

Trn ph 13C-NMR xut cc hin tn hiu ca mt cacbon gn vi nit ti
C 53,29; 3 nhm metyl ti C 15,96 v 14.13 (x2); 4 cacbon olefin ti C 136,19,
134,21, 128,76 v 123,27; 2 cacbon gn vi xy ti C 72,29 v 73,44. Thm vo
, 6 tn hiu ca mt phn t ng -D-glucopyranose xut hin ti C 103,10
(C-1), 77,44 (C-5), 76,32 (C-3), 76,15 (C-2), 69,71 (C-4) v 61,35 (C6).
T cc phn tch nu trn, c th s b xc nh SH3 l mt
glucocerebroside c 2 nhm hydroxyl, 2 lin kt i (mt b th 2 ln v 1 b th
3 ln).
Bng III.2.4.3.Kt qu ph cng hng t ht nhn ca SH3
C

Ca, b [215]

Cb, c

1
2
3
4
5
6
7
8
9
10
11-15
16
17

70,1 t
54,5 d
72,3 d
131,8 d
132,3 d
33,0 t
28,1 t
124,1 d
136,0 s
39,9 t
29,5-29,9 t
32,0 t
22,9 t

68,54 t
53,29 d
72,29 d
128,76 d
134,21 d
32,69 t
28,13 t
123,27 d
136,19 s
39,79 t
29,43-29,79 t
31,98 t
22,75 t

244

Hc, d
mult. (J in Hz)
3,76/4,03
3,84 dd (2,5, 12,0)
4,11 t (5,5)
5,46 dd (6,5, 15,5)
5,74 dd (6,5, 15,5)
2,05 m
2,05 m
5,10
1,95 m
1,26 br s
1,26 br s
1,26 br s

18
19
1
2
3
4
5-17
18
19
20
1
2
3
4
5
6
a

14,2 q
16,0 q
72,4 d
35,6 t
25,8 t
29,5-29,9 t
32,0 t
22,9 t
14,2 q
105,6 d
75,0 d
78,4 d
71,4 d
78,5 d
62,6 t

14,13 q
15,96 q
176,05 s
73,44 d
34,57 t
25,33 t
29,43-29,79 t
31,98 t
22,75 t
14,13 q
103,10 d
76,15 d
76,32 d
69,71 d
77,44 d
61,35 t

0,88 t (6,5)
1,57 s
4,03
1,76/1,54
1,38 m
1,26 br s
1,26 br s
1,26 br s
0,88 t (6,5)
4,27 d (7,5)
3,25 m
3,42 m
3,44 m
3,28 m
3,75 m/4,01 m

o trong Pyridine, b 125 MHz, co trong CDCl3 , d500 MHz

Hnh III.2.4.3.e. Ph 13C-NMR v cc ph DEPT ca SH3

Tn hiu singlet ti H 1,47 cho thy mt nhm metyl phi c ni vo
mt ni i v gi tr ti C 15,96 gi cho cu trc hnh hc dng trans ca ni
i ny [213]. Ni i cn li cng c xc nh c cu hnh trans bi hng s
tng tc ln (J = 15,5 Hz) ca cc proton. Bng vic so snh s liu ph NMR
ca SH3 vi hp cht c cu trc tng t [215] c th khng nh nhnh baz
mch di ca n l 2-amino-9-methyl-4,8-octadecadiene-1,3-diol. Xem xt k

245

lng gi tr chuyn dch ha hc ti cc v tr ca n v ng cho thy, ti

v tr C-2 c s chuyn dch rt mnh v vng trng thp. iu ny gi cho s
c mt ca nhm sulphonyl v tr ny [216]. T cc phn tch trn, cng vi
kt qu ph khi lng ESI-MS: [m/z 848.6 [M+H]+ (postitive), 846.8 [M-H](negative)] c th xc nh c nhnh axit bo l 2-hydroxyeicosanoic acid. Cu
trc lp th ti cc v tr C-2, 3 v 2 c xc nh bng vic so snh s liu ph
1
H- v 13C-NMR vi hp cht c cu trc tng t [215, 216]. Nh vy cu trc
ton b ca SH3 c xc nh l: 1-O-(2-sulfo--D-glucopyranosyl)(2S,3R,4E,8E)-2-[(2R)-2-hydroxyeicosanoylamino]-9-methyl-4,8-octadecadiene1,3-diol. y cng l mt hp cht mi v c chng ti t tn l
cladiacerebroside.

Hnh III.2.4.3.f. Ph ESI-MS ca SH3

III.2.4.4. Hp cht SH4: 3,6,11-trihydroxy-24-methylene-9,11-secocholest-7en-9-one
Hp cht SH4 nhn c di dng tinh th hnh kim mu trng, Cng
thc phn t ca n c xc nh l C28H46O4 bng cc d kin ph khi lng
(positive ESI-MS: m/z 447.2 [M + H]+, negative ESI-MS: m/z 445.4 [M - H]-) v
ph cng hng t ht nhn.

246

Hnh III.2.4.4.a. Ph 1H-NMR ca SH4

28
21

HO

11
18
12

19

20
17

26

24

13

25

27

14
9
8

10

H
3

7
6

HO
H

OH

Hnh III.2.4.4.b: Cu trc ho hc ca SH4

Ph 1H-v 13C-NMR ca SH4 tng t nh ca SH1, ngoi tr s mt i
cc tn hiu ca n v ng fucose. Nh vy c th xc nh SH4 cng l mt
hp cht 9,11-secosterol.

247

Hnh III.2.4.4.b: Ph 13C-NMR ca SH4

Hnh III.2.4.4.c: Cc ph DEPT ca SH4

248

S thay i ng ch nht trn ph 13C-NMR ca hp cht ny so vi

hp cht SH1 l v tr ca nhm carbonyl v ni i nm trong khung: gi tr
chuyn dch ha hc ca nhm carbonyl tng t C 205,1 ln 217,3 v ni
i gim t C 137,3/147,2 xung 121,9/139,6. S thay i ny cho thy v tr
ca lin kt i b thay i. V tr ca lin kt i c xc nh ti C-5/C-6
bi s ph hp hon ton v s liu ph 1H-v 13C-NMR ti cc v tr tng ng
ca SH4 so vi cc hp cht 9,11-secosterol c cu trc tng t [217, 218].
Ngoi ra s mt i tn hiu cng hng ca nhm oxymetin C-16 ti C 72,0 trn
ph 13C-NMR v thay vo l tn hiu ca nhm metylen ti C 26,3 khng
nh s mt i nhm hydroxyl ti v tr C-16 ca hp cht ny so vi SH1. Cc v
tr nhm th v cu hnh lp th ca cc v tr cn li ca SH4 hon ton trng
khp so vi SH1.

Hnh III.2.4.4.d: Ph HSQC ca SH4

Bng III.2.4.4. Kt qu ph NMR ca SH4
C

Ca, b

1
2
3
4
5
6

32,07 t
30,71 t
70,12 d
33,00 t
48,86 d
69,54 d

Ha, c
(mult., J in Hz)
1,46/1,90
1,45/1,92
3,58 (m)
1,42/2,30
1,76
4,26 (d, 8,0)

249

HMBC (H C)

7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28

147,16 d
137,33 s
205,07 s
45,00 s
59,45 t
41,35 t
46,39 s
42,85 d
26,88 t
26,28 t
50,09 d
17,49 q
16,21 q
34,82 d
18,99 q
34,32 t
31,84 t
156,70 s
33,98 d
21,92 q
22,05 q
106,34 t

6,56 (d, 1,5)

3,65 (dd, )/3,85 (dd, )
1,12/1,62
3,42 (t, 10,5)
1,60
1,45/1,88
1,75
0,66 (s)
1,13 (s)
1,45
0,99 (d, 6,5)
1,17/1,58
1,92/2,12
2,23
1,03 (d, 6,5)
1,03 (d, 6,5)
4,66 (s)/4,73 (s)

C-5, 9, 14

C-11, 18
C-7, 8, 9, 13, 15, 18

C-12, 13, 17
C-1, 5, 9, 10
C-17, 20, 22

C-24, 26, 27
C-24, 25, 27
C-24, 25, 26
C-23, 25

o trong CDCl3+MeOD, b 125 MHz, c500 MHz.

Hnh III.2.4.4.e: Ph HMBC ca SH4

250

Hnh III.2.4.4.f: Ph ESI-MS ca SH4

28
21

HO

11
20

18
12
19

17

13

26

24
25

27

14
9
8

10
3

5
6

HO
H

OH

Hnh III.2.4.4.g. Mt s tng tc HMBC chnh ca SH4

T cc phn tch nu phn trn, cng vi kt qu ph khi lng
(ESI-MS) v vic phn tch chi tit cc tng tc trn ph HMBC ca SH4 (Bng
III.2.4.4 v Hnh III.2.4.4.g) cho php xy dng v xc nh cu trc chnh xc l
3,6,11-trihydroxy-24-methylene-9,11-secocholest-7-en-9-one
(Hnh
III.2.4.4.b). Cu trc ny mt ln na c khng nh bi s trng khp v s
liu ph 1H- v 13C-NMR ca SH4 so vi hp cht trn c cng b [219].
Hp cht ny c phn lp t loi hi min Spongia officinalis v loi san h
mm Sinularia hirta. Tuy nhin y l ln u tin n c phn lp t chi
Cladiella.
251

III.2.4.5. Hp cht SH5: Cholesterol

Hp cht SH5 nhn c di dng tinh th hnh kim c mu trng. Th
nghim trn sc k lp mng vi thuc th c hiu (phun dung dch H2SO4 10%
ri h nng t t cho mu tm hng, sau chuyn dn sang tm xanh) cho thy
y l mt steroit.

Hnh III.2.4.5.a. Ph 1H-NMR ca SH5

Ph 1H-NMR ca SH5 c trng cho mt hp cht steroit vi s xut hin
cc tn hiu ca 5 nhm metyl ti 1,00 (3H, s, H-19), 0,91 (3H, s, H-21), 0,86
(6H, d, J = 6.5 Hz), H-26, 27) v 0.68 (3H, s, H-18), tn hiu ca proton olefin ti
5,35 (1H, br d, J = 5,0 Hz, H-6), v tn hiu ca proton gn vi cacbon c ni
vi nguyn t oxi ti 3,52 (1H, m, H-3).
21
20
18
17
19

11

22
25

27

14
8

HO

5
6

Hnh III.2.4.5.b: Cu trc ho hc ca SH5

252

26

Hnh III.2.4.5.c: Ph 13C-NMR ca SH5

Hnh III.2.4.5.d: Cc ph DEPT ca SH5

Ph 13C-NMR ca SH5 xut hin 27 tn hiu cacbon. Trn cc ph DEPT
xut hin cc tn hiu ca: 11 nhm CH2 ti 37.2 (t, C-1), 31.6 (t, C-2), 42,2 (t,
C-4), 31,8 (t, C-7), 21,1 (t, C-11), 39,8 (t, C-12), 24,3 (t, C-15), 28,3 (t, C-16),
36,1 (t, C-22), 23,8 (t, C-23) v 39,5 (t, C-24); 5 tn hiu CH3 ti 22,5 (q, C26), 22,8 (q, C-27). 11,9 (q, C-18), 19,4 (q, C-19) v 18,7 (q, C-21) ; 8 tn hiu
CH ti 71.8 (d, C-3), 121,7 (d, C-6), 31,8 (d, C-8), 50,1 (d, C-9), 35,7 (d, C253

20), 28,0 (d, C-25), 56,7 (d, C-14), 56,1 (d, C-17) v 3 tn hiu C ti 140,7 (s,
C-5), 36,5 (s, C-10), 42,3 (s, C-13). Trong mt ni i c xc nh bi hai
tn hiu ti 140,7 (C) v 121,7 (CH), mt cacbon c ni vi nguyn t oxi ti
71,8 (d, C-3) c trng cho cu hnh 3-OH. Nhng d kin ph trn gi ti
mt hp cht cholesterol, mt steroit rt ph bin ng vt. T nhng phn
tch nu trn, s liu ph 13C-NMR c so snh trc tip vi cc s liu ph ca
hp cht cholesterol [220]. S ph hp hon ton v cc gi tr ph tng ng
cho php khng nh hp cht SH5 l cholesterol.
III.2.4.6. Hp cht SH6: Chimyl alcohol
Hp cht SH6 nhn c di dng kt tinh mu trng, ph khi lng
ESI-MS m/z: 317,1 [M+H]+, (C19H40O3, M=316).

Hnh III.2.4.6.a. Ph 1H-NMR ca SH6

3

HO

OH

1'

16'

CH2(CH2)14CH3

Hnh III.2.4.6.b. Cu trc ha hc ca SH6

Trn ph 1H-NMR ca SH6 xut hin cc tn hiu c trng ca cc nhm
oximetin v oximetilen ti 3,86 (1H, m), 3,71(1H, dd, J = 5,0, 11,0 Hz), 3,64
(1H, dd, J = 3,0, 11,0 Hz), 3,52 (2H, m) v 3,45 (2H, m); cc proton ca cc

254

nhm OH ti 2,64 (1H, br s) v 2,17 (1H, br s); mt nhm metyl ti 0,88 (3H,
s) v cc nhm metilen ti 1,57 (2H, m) v 1,26 (28H, br s).

Hnh III.2.4.6.c. Ph 13C-NMR ca SH6

Bng III.2.4.6. Kt qu ph NMR ca SH6.
C

C [221]

71,8

2
3
1
2
3
4-13
14
15
16
2-OH
3-OH

70,5
64,2
72,4
29,0
26,1
29,3-29,7
31,9
22,7
14,1
-

C a, b

Ha, c (J, Hz)

72,54 (t)
70,45 (d)
64,33 (t)
71,89 (t)
29,30-29,71 (t)
26,10 (t)
29,30 - 29,71 (t)
31,94 (t)
22,70 (t)
14,12 (q)
-

3,64 (dd, 3,0, 11,0)

3,71 (dd, 5,0, 11,0)
3,86 (m)
3,52 (m)
3,45 (m)
1,58 (m)
1,26 (br s)
1,26 (br s)
1,26 (br s)
1,26 (br s)
0,88 (t, 7,0)
2,17 * (br s)
2,64 * (br s)

o trong CDCl3, b125MHz, c 500MHz, *gi tr c th i ch cho nhau.

Trn ph 13C-NMR xut hin cc tn hiu c trng ca mt mch di

cacbon no ti 31,94 (t), 29,71 (t), 29,67 (t), 29,62 (t), 29,60(t), 29,47 (t), 29,37
(t), 26,10 (t), 22,70 (t) v 14,12 (q). Cc tn hiu ca mt gc glyxerin b th mt
v tr v khng i xng -CH2OCH2CH(OH)CH2OH cng c khng nh bi s
xut hin cc tn hiu cacbon ti 72,54 (t, C-1), 70,45 (d, C-2), 64,33 (t, C-3) v
71,89 (t, C-1) [221]. S ph hp v gi tr quay quang ca SH6 vi (S)255

Chimyl cng vi vic tt c cc cc te glyxerit phn lp c cho n nay u

dng cu hnh (S) [221, 222] cho php d on cu hnh ti v tr C-2 l dng
(S).

Hnh III.2.4.6.d. Ph 13C-NMR v cc ph DEPT ca SH6

Cc phn tch nu cho php khng nh SH6 l mt glyxerol mono
alkyl te. T , s liu ph NMR ca n c mang so snh vi cc s liu
c cng b ca 1-O-hexadecyl-glycerol (chimyl alcohol) [221] v thy s ph
hp hon ton cc v tr tng ng.
Ngoi ra s xut hin pc ion phn t ti m/z 317.1 trn ph khi lng
ESI-MS (tng ng vi cng thc phn t C19H40O3, M=316) cho php xc nh
chnh xc di ca mch cc bon no v cu trc ha hc ca SH6 l chimyl
alcohol (glycerol 1-O-hexadecyl ether).

Hnh III.2.4.6.g. Ph khi lng ESI-MS ca SH6

256

III.2.4.7. Hp cht SH7: Batyl alcohol

Hp cht SH7 cng nhn c di dng tinh th mu trng. Ph 1HNMR ca SH7 (Hnh III.2.4.7.a) gn nh trng khp hon ton vi SH6 ngoi tr
gi tr tch phn ca pc cng hng ti 1,26 (30H, br s) ln hn so vi SH6.

Hnh III.2.4.7.a. Ph 1H-NMR ca SH7

Nh vy c th d on rng cu trc ha hc ca hp cht ny hon ton
tng t nh ca SH6 v ch c mch cacbon no di hn m thi. khng nh
chnh xc s cacbon tng thm, ph khi lng (ESI-MS) c o. Vic xut
hin pc ion phn t ti m/z 345,1 trn ph positive ESI-MS cho php khng nh
hp cht ny c di mch cacbon no ln hn so vi SH-6 l 2 cacbon. T tt
c cc phn tch nu, SH7 c nhn dng l batyl alcohol (glycerol 1-Ooctadecyl ether) [221].
3

HO

OH

1'

18

CH2(CH2)16CH3

Hnh III.2.4.7.b. Cu trc ha hc ca SH7

Hnh III.2.4.7.e. Ph khi lng ESI-MS ca SH7

257

III.2.4.8. Hp cht SH8: 1-Heptadecanol

Hp cht SH8 nhn c di dng cht kt tinh mu trng. Ph 1H-NMR
xut hin cc tn hiu c trng ca 1 nhm metyl ti 0,88 (3H, t, J = 7 Hz); 16
nhm metilen ca mt cc bon no mch di ti 1,26 (30H, br s) v 1,57 (2H,
m); v mt nhm oxymetilen ti 3,63 (2H, t, J = 7,0 Hz)

Hnh III.2.4.8.a. Ph 1H-NMR ca SH8

1

16
17

HO

14

CH3

Hnh III.2.4.8.b. Cu trc ha hc ca SH8

Vic xut hin tn hiu duy nht ca 1 nhm metyl v tn hiu ca 1 nhm
oxymetilen cng vi s tch vch b 3 vi hng s tng tc J = 7 Hz cc tn
hiu ny cho php khng nh 2 nhm ny u lm tn cng ca mch cacbon
no v mch ny khng r nhnh. Cng vi kt qu ph khi lng (postitive ESIMS m/z 279.0 [M+Na]+, negative ESI-MS m/z 255.0 [M-H]-, 237.0 [M-H2O-H]tng ng vi cng thc phn t C17H36O) cho php khng nh cu trc ha hc
ca SH8 l 1-heptadecanol.

258

Hnh III.2.4.8.c. Cc ph ESI-MS ca SH8

259

III.2.5. Kt qu nh gi hot tnh sinh hc ca cc cht

III.2.5.1. Kt qu nh gi hot tnh khng NF-B ca cc hp cht
Cc hp cht phn lp c t loi san h mm Cladiella sp c tin
hnh th nghim hot tnh khng NF-B, kt qu th nghim c th c nu ra
trong bng III.2.5.
Bng III.2.5.1. Kt qu th nghim hot tnh khng NF-B
Hp cht

IC50 (M)

Parthelolide (chng dng)

SH1
SH2
SH3
SH4
SH5
SH6
SH7
SH8

6,66 0,07
>50
14,8 1,73
>50
32,5 1,80
>50
>50
>50
>50

Kt qu trn cho thy, ch c hai hp cht l SH2 v SH4 th hin hot

tnh khng NF-B. iu ny cho thy s ph hp so vi cc ti liu cng b v
SH2 l mt hp cht ceramit cn SH4 l mt 9,11-secosterol, hai lp cht th
hin rt nhiu hot tnh qu, cn c quan tm nghin cu pht trin thnh
cc ch phm thuc c gi tr cao, phc v cng tc chm sc sc khe cng
ng. Hp cht SH1 cng l mt 9,11-secosterol tuy nhin n li dng dn xut
glycosit, v mt iu th v l y l dn xut glycosit u tin ca cc hp cht
9,11-secosteroit c phn lp t thin nhin. Hp cht ny khng th hin hot
tnh khng NF-B c th l do s nh hng ca n v ng fucose trong phn
t.
III.2.5.2. Kt qu nh gi hot tnh gy c t bo ca cc hp cht
Kt qu th nghim (bng III.2.5.2.) cho thy ch c 4 hp cht th hin
hot tnh gy c t bo. Trong hp cht SH1 ch c hot tnh gy c t bo
i vi dng KB vi IC50 = 4,43 g/ml, SH2 v SH4 th hin hot tnh trn c 3
dng t bo th nghim, SH3 th hin hot tnh trn 2 dng t bo l KB v HepG2 vi gi tr IC50 = 3,77 v 4,72 g/ml. Mt iu ng ch l hp cht SH2 l
mt hp cht mi, th hin hot tnh gy c t bo trn c 3 dng t bo th
nghim v c phn lp vi hm lng tng i cao t loi san h mm
Cladiella sp. iu ny gi m cho cc nghin cu tip theo ca hp cht ny theo
nh hng hot tnh gy c t bo nhm pht trin cc loi dc liu mi c tc
dng phng v iu tr bnh ung th.

260

Bng III.2.5.2. Hot tnh gy c t bo ca cc cht

Hp cht
DMSO
Ellipthine
SH1
SH2
SH3
SH4
SH5
SH6
SH7
SH8

Gi tr CS% (Cell survival)

IC50 (g/ml)

KB

FL

Hep-G2

KB

FL

Hep-G2

1000,0

1000,0

100,00,0

1,071,07
45,60,5

1,340,63
74,21,5
39,21,1

0,21

0,12

0,19

32,12,6

1,520,00
81,5,10,7
44,22,5

39,40,7
41,20,9

74,22,2

46,70,9

31,11,2

24,21,6

73,81,2

77,70,9

65,21,2

82,22,1

69,90,6

56,70,8

69,43,5

74,52,4

86,31,5

92,61,7

81,31,8

61,10,5

4,43
3,37
3,77
4,23
(-)
(-)
(-)
(-)

(-)
4,06
(-)
2,97
(-)
(-)
(-)
(-)

(-)
3,69
4,72
1,95
(-)
(-)
(-)
(-)

III.2.5.3. Hot tnh chng xy ha

Kt qu nh gi hot tnh chng xy ha thng qua phn ng bao vy gc
t do DPPH cho thy, ch c hp cht SH1 v SH3 th hin hot tnh vi gi tr
ED50 tng ng l 77,8 v 65,5 g/ml.
Bng II.2.5.3. Hot tnh chng xy ha thng qua kh nng thu dn gc t do
DPPH ca cc hp cht.
Hp cht
Chng (-)
Chng (+)
SH1
SH2
SH3
SH4
SH5
SH6
SH7
SH8

Gi tr SC %
(scavenging capacity)
0,000,00
80,920,42
57,780,59

ED50 (g/ml)
>100
19,8
77,8
(-)
65,5
(-)
(-)
(-)
(-)
(-)

44,451,12

65,410,67
47,110,92
33,211,65
38,752,23
24,511,80
34,332,35

T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc

loi san h mm Cladiella sp, chng ti bin son v ng ti 01 cng trnh
trn tp ch chuyn ngnh quc t l:
-

Chau Van Minh, Phan Van Kiem, and Nguyen Xuan Cuong, New 9,11-secosterol
glycoside and ceramide from a soft coral Cladiella sp. living in Vietnamese sea. ASEAN
Journal on Science and Technology for Development, 2007, 24(3), 275-283.

261

Bng tng hp cc cht phn lp c t loi

san h mm cladiella sp.
28

20

11

24

23

22

25

18

12
19
6'
O
1'

4'

HO

3'

10

8
6

2'

13
14

1'

3'

HN

OH

15

16'

10

27

17
16

1
5'

OH

26

21

HO

18

HO

OH
OH

HO

SH2: Cladiaceramide (Cht mi)

SH1: Cladiasterol (Cht mi)

OH
O

28

1'

4"

6"

HO
HO

18

5"
2"

22

23

24

1"

SO3H

14
2

19

15

10
5

HO

27

17
16

HO

13

19

25

18
12

3"

20

11

HN

OH

26

21

HO

20'

16

7
6

SH3: Cladiacerebroside (Cht mi)

OH

SH4: 3,6,11-trihydroxy-24methylene-9,11-secocholest-7-en-9-one
21
20

21

24

HO

17

26

19

8
6

HO

1'

CH2(CH2)nCH3

SH6: Chimyl alcohol n = 14

SH7: Batyl alcohol n = 16

SH5: Cholesterol

16
17

HO

OH

2
1

25

18

14

CH3

SH8: 1-Heptadecanol

262

Chng IV. Nghin cu ho hc theo nh hng

hot tnh sinh hc ca mt s loi da gai.
IV.1. nghin cu ha hc theo nh hng hot tnh sinh
hc loi hi sm holothuria scabra (holothuridae)
IV.1.1. M u
Mu hi sm Holothuria scabra c thu thp ti Ct B, Hi Phng vo
thng 2 nm 2006 v thng 8 nm 2007 v c TS. Cng Thung v cc cng
s, Vin Ti nguyn v Mi trng Bin, Vin KH&CN Vit Nam gim nh tn
khoa hc.
Tn Vit Nam: Hi sm trng
Tn gi khc : Da chut bin
Tn khoa hc: Holothuria scabra
a im ly mu: Vng bin Ct b.
Ni sng: Sinh sng Khu vc Bi Hng
Vn n hn Khe Tru. Hi sm thng
sng trn cc rn san h v theo nc
triu tp vo bi ct kim n.

Hi sm trng (Holothuria scabra) l loi ng vt khng xng sng

thng gp vnh v nhng ni c nhiu ngm bin khi. Hi sm phn b
nhiu nc, trong c nc ta. Theo kinh nghim y hc c truyn hi sm c
xem nh loi thuc b thn, b m, trng dng ch tinh, nhun to, cha l, cha
vim ph qun, thn kinh suy nhc, cm mu. Ngy nay, vi s tr gip ca cc
phng tin khoa hc v k thut hin i, vic nghin cu cc ch tiu bn trong
ca hi sm khng nhng lm sng t cc kinh nghim s dng hi sm lm dc
liu trong dn gian m cn gip tm ra cc phng thuc cha bnh t loi sinh
vt bin ny. Cc nghin cu v thnh phn ha hc c tin hnh cho thy,
thnh phn ha hc chnh ca chi Holothuria l cc sterol v sterol glycosit (cc
holothurin) c khung c bn l holostane. Hng trm hp cht thuc dng khung
ny vi nhiu hot tnh ng quan tm c phn lp t cc loi hi sm.
Holothuria scabra l mt loi sinh sng kh ph bin ti cc vng bin vit nam.
Hin c rt t cc nghin cu v thnh phn ha hc v hot tnh sinh hc ca loi
ny trn th gii v cha c nghin cu no c tin hnh nc ta. Di y
chng ti gii thiu cc hp cht c phn lp v xc nh cu trc t loi
ny.
O

O
OH

HO

HO
H

9,11-Dihydro-22,25-oxido-11oxoholothurinogenin [223]

4,14 -Dimethylcholest-7-en-3-ol [224]

263

H
H
H
HO
O
O

HO

HO
OH

24,26-Dimethylcholest-5-en- 3-O--DXylopyranoside [225]

O

4,14 ,24-Trimethylcholesta-7,22-dien-3ol [224]

H3CO
OH

OH

HO

HO
H

12-Methoxyholost-9(11)-ene-3,17-diol [223]

3,17-Dihydroxylanosta-7,9(11)-dien18,20-olide [223]
O

HO
OH
H

RO

HO

R = 3-O-[3-O-methyl--D-glucopyranosyl-(13)-D-glucopyranosyl-(14)-6-deoxy--Dglucopyranosyl-(12)-4-O-sulfo--Dxylopyranoside] 24-Dehydroechinoside A [226]

Dihydroparkeol [224]

IV.1.2. Phn lp cc hp cht

Mu Holothuria scabra (10kg kh) c ra sch nhiu ln loi mui
v c, sau thi nh v chit siu m bng methanol. Qu trnh phn lp cc
hp cht c trnh by c th s di y, kt hp gia cc phng php
sc k ct Silicagen pha thng, pha o YMC v sc k lp mng TLC. T 200
gam dch c methanol, em chit phn b bng hexan v nc vi t l 1/1 thu
c 80 gam dch chit. Tng t ln lt chit vi cloroform, butanol v nc
theo t l 1/1 v th tch thu c 70 gam dch chit cloroform v 30 gam dch
chit butanol, phn cn nc cn li l 20 gam. Phn dch chit CHCl3 c tm
vo 90 gam Silicagel, sau a ln ct v chy sc k vi hn hp dung mi ngexan/axeton c gradient phn cc tng dn t 0% n 100% axeton, thu c
6 phn on khc nhau. Phn on s 3 tip tc c chy sc k ct Silicagel
pha thng vi h dung mi n-hexan/axeton 4:1 thu c 2 cht sch HS-7 (15
mg) v HS-8 (12 mg) u c dng tinh th hnh kim, mu trng. Nhm 7 cng
c chy sc k ct pha thng vi h n-hexan/axeton 10:1 thu c cc tinh
th hnh kim mu trng HS-3 vi lng 10 mg. T nhm 5 sau khi tin hnh sc
k ct pha thng bng h dung mi CHCl3/MeOH 2,3:1 thu c lng ln hai
cht bt mu trng, k hiu l HS-1 v HS-2. Nhm 4 cng c tin hnh sc k
264

nh nhm 5, thu c 2 cht HS-6 dng bt mu trng (S IV.1.2.a.). Bng

cc phng php tng t, cc hp cht HS-3 v HS-9 c phn lp t cn
butanol, HS-4 v HS-5 t cn nc.
Hi sm Holothuria
scabra (10 kg)
Thi nh, chit siu m trong
MeOH, ct loi dung mi
Dch chit MeOH
(200 g)
B sung nc
B sung CHCl3

Chit n-Hexan 1:1

Chit CHCl3 1:1

B sung n-BuOH

Chit BuOH 1:1

Cn n-Hexan
(80 g)

Cn CHCl3
(70 g)

Cn n-BuOH
(30 g)

CC, Sephadex
CHCl3/MeOH/H2O
1:4:1

CC, silicagel, n-hexan/axeton

gradien phn cc tng dn

10

12

HS-4 (9 mg)

HS-9 (12 mg)

HS-3 (10 mg)

265

11

CC, Silicagel, YMC, HS-5 (13 mg)

Axeton/H2O 1/1

CC, Silicagel, YMC,

MeOH/H2O 1/3

CC, Silicagel, YMC,

Axeton/H2O 1/1

HS-6 (9 mg)

CHCl /MeOH 2/1

HS-1
(250 mg)
HS2 (220 mg)
3/

CHCl3//MeOH 9/1

n-hexan/axeton 4/1

HS-7 (15 mg)

HS-8(12 mg)

S IV.1.2. S qu trnh phn lp cc hp cht t

hi sm Holothuria scabra

100% MeOH

CC, Dianion
MeOH/H2O
70% MeOH

20% MeOH

100% axeton

80% axeton

40% n-hexan

60% n-hexan

80% n-hexan

100% n-hexan

Cn pha nc
(20 g)

IV.1.3. Hng s vt l v d kin ph ca cc hp cht

IV.1.3.1. Hp cht HS1: Holothurin B
Cht bt mu trng, im chy 251-253oC, ph khi lng (positive ESI-MS) ti
m/z 905 [M+Na]+, cng thc phn t ca C41H63O17NaS (M = 882).
1
H-NMR (500 Hz) : 3,12 (1H, dd, J = 10,0, 3,0 Hz, H-3), 3,40 (1H, m, H-8),
5,40 (1H, d, J = 5,0, H-11), 1,13 (3H, s, H-19), 1,52 (3H, s, H-21), 1,12 (3H, s, H-26),
1,03 (3H, s, H-27), 0,89 (3H, s, H-28), 1,28 (3H, s, H-29), 1,50 (3H, s, H-30), 4,60 (1H,
d, J = 7,0 Hz, H-1), 4,68 (1H, d, 7,8 Hz, H-1), v 1,72 (3H, d, J = 6,0 Hz, H-6).
13
C-NMR (125 MHz) : 36,3 (t, C-1), 27,6 (t, C-2), 89,5 (d, C-3), 40,0 (s, C-4),
52,8 (d, C-5), 21,1 (t, C-6), 27,9 (t, C-7), 40,8 (d, C-8), 153,7 (s, C-9), 39,7 (s, C-10),
114,2 (d, C-11), 71,3 (d, C-12), 59,3 (s, C-13), 46,0 (s, C-14), 28,1 (t, C-15), 38,5 (t, C16), 89,7 (s, C-17), 174,9 (s, C-18), 18,4 (q, C-19), 86,0 (s, C-20), 22,5 (q, C-21), 80,1
(d, C-22), 36,8 (t, C-23), 28,7 (t, C-24), 82,5 (s, C-25), 28,7 (q, C-26), 28,1 (q, C-27),
21,0 (q, C-28), 27,4 (q, C-29), 16,7 (q, C-30), 104,5 (d, C-1), 82,2 (d, C-1), 75,6 (d, C3), 73,3 (d, C-4), 62,9 (t, C-5), 104,5 (d, C-1), 75,6 (d, C-2), 76,0 (d, C-3), 75,9 (d,
C-4), 72,8 (d, C-5), v 17,8 (q, C-6).
IV.1.3.2. Hp cht HS2: Holothurin A
Cht bt mu trng, im chy 228-230oC.
1
H-NMR (500 Hz, CD3OD) : 3,12 (1H, dd, J = 10,0, 3,0 Hz, H-3), 3,40 (1H, m,
H-8), 5,40(1H, d, J = 5,0 Hz, H-11), 1,13 (3H, s, H-19), 1,52 (3H, s, H-20), 1,12 (3H, s,
H-26), 1,03 (3H, s, H-27), 0,89 (3H, s, H-28), 1,28 (3H, s, H-29), 1,50 (3H, s, H-30),
4,30(1H, d, J = 7,0 Hz, H-1), 4,50(1H, d, J = 7,8 Hz, H-1), 1,72(3H, d, J = 6,0 Hz, H6), 4,30(1H, d, J = 7,0 Hz, H-1), 4,30(1H, d, J = 7,0 Hz, H-1), v 3,68 (3H, s, H7).
13
C-NMR (125 MHz, CD3OD) : 35,8 (t, C-1), 27,4 (t, C-2), 90,4 (d, C-3), 40,6
(s, C-4), 53,5 (d, C-5), 21,8 (t, C-6), 28,8 (t, C-7), 41,7 (d, C-8), 155,6 (s, C-9), 40,4 (s,
C-10), 115,2 (d, C-11), 72,3 (d, C-12), 59,7 (s, C-13), 46,4 (s, C-14), 28,6 (t, C-15), 39,0
(t, C-16), 90,3 (s, C-17), 176,6 (s, C-18), 19,0 (q, C-19), 88,0 (s, C-20), 22,7 (q, C-21),
81,3 (d, C-22), 37,4 (t, C-23), 28,6 (t, C-24), 83,3 (s, C-25), 28,7 (q, C-26), 28,5 (q, C27), 20,4 (q, C-28), 27,5 (q, C-29), 17,1 (q, C-30), 105,4 (d, C-1), 82,1 (d, C-2), 77,1
(d, C-3), 74,3 (d, C-4), 63,9 (t, C-5), 104,9 (d, C-1), 75,5 (d, C-2), 76,1 (d, C-3),
86,4 (d, C-4), 72,2 (d, C-5), 18,1 (q, C-6), 104,7 (d, C-1), 75,1 (d, C-2), 87,1 (d,
C-3), 70,8 (d, C-4), 77,3 (d, C-5), 62,2 (t, C-6), 104,3(d, C-1), 74,9 (d, C-2),
86,7 (d, C-3), 69,6 (d, C-4), 77,6 (d, C-5), 62,1 (t, C-6), v 61,1 (q, C-7).
IV.1.3.3. Hp cht HS3: Holothurin A2
Cht bt mu trng, c im chy 246-2480 C, []D20: -11,050 (C=0,26; C2H5OH),
khi lng positive ESI-MS: m/z 1229,6 [M+Na]+, cng thc phn t: C54H87O26SNa.
1
H-NMR (500 Hz, DMSO) : 1,37 - 1,75 (2H, m, H-1), 1,64 - 1,87 (2H, m, H2), 3,39 (1H, m, H-3), 0,88 (1H, m, H-5), 1,67 (2H, m, H-6), 1,35 - 1,67 (2H, m, H-7),
2,88 (1H, dd, J = 4,5, 5,0 Hz, H-8), 5,26 (1H, d, J = 4,0 Hz, H-11), 4,43 (1H, d, J = 5,0
Hz, H-12), 1,05 - 1,70 (2H, m, H-15), 1,95 - 2,41 (2H, m, H-16), 1,03 (3H, s, H-19),
1,43 (3H, s, H-21), 1,65 (2H, m, H-22), 1,31 - 1,40 (2H, m, H-23), 1,18 (2H, m, H-24),

266

1,55 (1H, m, H-25), 0,87 (3H, s, H-26), 0,86 (3H, s, H-27), 0,81 (3H, s, H-28), 1,02
(3H, s, H-29), 1,20 (3H, s, H-30), 4,32 (1H, d, J = 7,0 Hz, H-1), 3,37 (1H, m, H-2),
3,08 (1H, m, H-3), 3,93 (1H, m, H-4), 3,21 - 3,95 (2H, m, H-5), 4,49 (1H, d, 8,0 J =
Hz, H-1), 3,52 (1H, m, H-2), 3,29 (1H, m, H-3), 3,00 (1H, m, H-4), 3,33 (1H, m,
H-5), 1,25 (3H, s, H-6), 4,38 (1H, d, J = 8,0 Hz, H-1), 3,21 (1H, m, H-2), 3,03
(1H, m, H-3), 3,21 (1H, m, H-4), 3,31 (1H, m, H-5), 3,41 - 3,68 (2H, m, H-6),
4,34 (1H, d, J = 8,0 Hz, H-1), 3,13 (1H, m, H-2), 3,00 (1H, m, H-3), 3,14 (1H,
m, H-4), 3,22 (1H, m, H-5), 3,41 - 3,68 (2H, m, H-6), v 3,49 (3H, s, -OCH3),
13
C-NMR (125 MHz, DMSO) : 35,8 (C-1), 26,1 (C-2), 87,5 (C-3), 38,9 (C-4),
51,9 (C-5), 20,3 (C-6), 27,1 (C-7), 40,0 (C-8), 152,6 (C-9), 39,8 (C-10), 114,5 (C-11),
70,1 (C-12), 57,4 (C-13), 45,3 (C-14), 35,6 (C-15), 34,7 (C-16), 88,13 (C-17), 173,6 (C18), 21,9 (C-19), 86,2 (C-20), 22,4 (C-21), 37,8 (C-22), 21,3 (C-23), 38,9 (C-24), 27,2
(C-25), 22,5 (C-26), 22,5 (C-27), 16,2 (C-28), 27,4 (C-29), 19,2 (C-30), 103,83 (C-1),
81,4 (C-2), 74,8 (C-3), 74,2 (C-4), 62,98 (C-5), 103,60 (C-1), 74,5 (C-2), 74,2 (C3), 86,1 (C-4), 70,10 (C-5), 17,3 (C-6), 102,92 (C-1), 72,1 (C-2), 87,9 (C-3),
68,4 (C-3), 76,07 (C-5), 60,74 (C-6), 103,9 (C-1), 73,5 (C-2), 85,9 (C-3),
69,26 (C-4), 76,9 (C-5), 60,85 (C-6), v 59,82 (-OCH3).
IV.1.3.4. Hp cht HS4: Holothurin A3
Cht bt mu trng, ph khi lng ESI-MS ti m/z 1259,3 [M + Na]+ (positive)
v 1235,5 [M - H]+, ph khi lng phn gii cao HR ESI-MS: m/z 1259,4900 [M +
Na]+ (positive) v kt qu tnh phn tch nguyn t cho cng thc C54H85O28SNa2 l
1259,4743, cng thc phn t C54H85O28SNa.
1
H-NMR (500 Hz, DMSO) : 1,41 - 1,74 (2H, m, H-1), 1,63 - 1,85 (2H, m, H-2),
3,34 (1H, m, H-3), 0,88 (1H, m, H-5), 1,67 (2H, m, H-6), 1,35 - 1,67 (2H, m, H-7), 2,87
(1H, m, H-8), 5,26 (1H, d, J = 4,5 Hz, H-11), 4,48 (1H, m, H-12), 1,69 - 2,05 (2H, m,
H-15), 1,70 (2H, m, H-16), 1,05 (3H, s, H-19), 1,56 (3H, s, H-21), 2,64 (2H, m, H-23),
1,54 - 1,61 (2H, m, H-24), 1,07 (3H, s, H-26), 1,07 (3H, s, H-27), 0,81 (3H, s, H-28),
1,02 (3H, s, H-29), 1,18 (3H, s, H-30), 7,03 (1H, s, OH-17), 4,23 (1H, s, OH-25), 4,32
(1H, d, J = 7,5 Hz, H-1), 3,34 (1H, m, H-2), 3,07 (1H, m, H-3), 3,94 (1H, m, H-4),
3,20 - 3,93 (2H, m, H-5), 4,49 (1H, d, J = 7,5 Hz, H-1), 3,51 (1H, m, H-2), 3,29 (1H,
m, H-3), 3,00 (1H, m, H-4), 3,33 (1H, m, H-5), 1,25 (3H, s, H-6), 4,38 (1H, d, J =
8,0 Hz, H-1), 3,21 (1H, m, H-2), 3,03 (1H, m, H-3), 3,21 (1H, m, H-4), 3,31
(1H, m, H-5), 3,41 - 3,68 (2H, m, H-6), 4,34 (1H, d, J = 7,5 Hz, H-1), 3,15 (1H,
m, H-2), 2,99 (1H, m, H-3), 3,14 (1H, m, H-4), 3,21 (1H, m, H-5), 3,41 - 3,68
(2H, m, H-6), 3,49 (3H, s, -OCH3), v 5,80 (1H, s, OH-3).
13
C-NMR (125 MHz, DMSO) : 35,7 (C-1), 26,2 (C-2), 87,5 (C-3), 38,9 (C-4),
51,9 (C-5), 20,6 (C-6), 27,4 (C-7), 40,0 (C-8), 152,8 (C-9), 39,7 (C-10), 114,5 (C-11),
69,5 (C-12), 57,3 (C-13), 45,2 (C-14), 37,8 (C-15), 35,8 (C-16), 91,3 (C-17), 172,6 (C18), 21,9 (C-19), 86,2 (C-20), 20,6 (C-21), 208,1 (C-22), 33,5 (C-23), 36,4 (C-24), 68,0
(C-25), 29,1 (C-26), 29,4 (C-27), 16,1 (C-28), 27,4 (C-29), 19,0 (C-30), 103,86 (C1),
81,4 (C-2), 74,8 (C-3), 74,2 (C-4), 63,0 (C-5), 103,62 (C-1), 74,5 (C-2), 74,2 (C3), 86,2 (C-4), 70,1 (C-5), 17,3 (C-6), 102,95 (C-1), 72,1 (C-2), 87,9 (C-3),

267

68,4 (C-4), 76,1 (C-5), 60,76 (C-6), 103,92 (C-1), 73,5 (C-2), 85,9 (C-3),
69,27 (C-4), 76,8 (C-5), 60,86 (C-6), v 59,87 (-OCH3).
IV.1.3.5. Hp cht HS5: Holothurin A4
Cht bt trng, ph khi lng ESI-MS (positive cho pic ion phn t m/z 1245,2
[M+Na]+ v negative cho pic ion m/z 1221,4 [M-H]-), cng thc phn t C54H87O27SNa
(M=1222).
1
H-NMR (500 Hz, DMSO) : 1,36 - 1,75 (2H, m, H-1), 1,65 - 1,85 (2H, m, H-2),
3,35 (1H, m, H-3), 0,86 (1H, m, H-5), 1,45 - 1,67 (2H, m, H-6), 1,34 - 1,67 (2H, m, H7), 2,99 (1H, m, H-8), 5,25 (1H, H-11), 4,43 (1H, m, H-12), 1,05 - 1,70 (2H, m, H-15),
1,93 - 2,45 (2H, m, H-16), 1,03 (3H, s, H-19), 1,42 (3H, s, H-21), 1,61 -1,92 (2H, m, H22), 1,22 - 1,40 (2H, H-23), 3,14 (1H, m, H-24), 1,55 (1H m, H-25), 0,83 (3H, s, H-26),
0,84 (3H, s, H-27), 0,81 (3H, s, H-28), 1,03 (3H, s, H-29), 1,20 (3H, s, H-30), 4,32 (1H,
d, J = 7,5 Hz, H-1), 3,34 (1H, m, H-2), 3,07 (1H, m, H-3), 3,94 (1H, m, H-4), 3,20 3,93 (2H, m, H-5), 4,49 (1H, d, J = 7,5 Hz, H-1), 3,51 (1H, m, H-2), 3,29 (1H, m, H3), 3,00 (1H, m, H-4), 3,33 (1H, m, H-5), 1,25 (3H, s, H-6), 4,38 (1H, d, J = 8,0
Hz, H-1), 3,21 (1H, m, H-2), 3,03 (1H, m, H-3), 3,21 (1H, m, H-4), 3,31 (1H, m,
H-5), 3,41 - 3,68 (2H, m, H-6), 4,34 (1H, d, J = 7,5 Hz, H-1), 3,15 (1H, m, H2), 2,99 (1H, m, H-3), 3,14 (1H, m, H-4), 3,21 (1H, m, H-5), 3,41 - 3,68 (2H,
m, H-6), 3,49 (3H, s, -OCH3), v 5,80 (1H, s, OH-3).
13
C-NMR (125 MHz, DMSO) : 35,67 (C-1), 26,2 (C-2), 87,57 (C-3), 38,33 (C4), 51,93 (C-5), 20,4 (C-6), 27,1 (C-7), 40,0 (C-8), 152,63 (C-9), 39,8 (C-10), 114,5 (C11), 70,15 (C-12), 57,46 (C-13), 45,38 (C-14), 35,7 (C-15), 34,8 (C-16), 88,2 (C-17),
173,7 (C-18), 21,9 (C-19), 86,6 (C-20), 22,53 (C-21), 34,3 (C-22), 28,2 (C-23), 74,8 (C24), 33,2 (C-25), 17,7 (C-26), 18,99 (C-27), 16,17 (C-28), 27,45 (C-29), 19,30 (C-30),
103,86 (C1), 81,4 (C-2), 74,8 (C-3), 74,2 (C-4), 63,0 (C-5), 103,62 (C-1), 74,5 (C2), 74,2 (C-3), 86,2 (C-4), 70,1 (C-5), 17,3 (C-6), 102,95 (C-1), 72,1 (C-2),
87,9 (C-3), 68,4 (C-4), 76,1 (C-5), 60,76 (C-6), 103,92 (C-1), 73,5 (C-2),
85,9 (C-3), 69,27 (C-4), 76,8 (C-5), 60,86 (C-6), v 59,87 (-OCH3).
IV.1.3.6. Hp cht HS6: Holothurigenin B
Cht bt v nh hnh, ph khi lng ESI-MS: positive ti m/z 762,7 [M+H]+,
785.3 [M+Na]+ v negative ti m/z 761.3 [M-H]-, cng thc phn t C41H62O13 (M =
762).
1
H-NMR (500 Hz, MeOD) : 2,8 (2H, m, H-1), 2,06 (2H, m, H-2), 3,15 (1H, m,
H-3), 1,15 (1H, m, H-5), 2,13 (2H, dd, J = 4,5, 6,5 Hz, H-6), 5,55 (2H, m, H-7), 5,30
(1H, t, H-11), 2,25 - 2,85 (2H, m, H-12), 1,36 - 1,86 (2H, m, H-15), 1,80 (2H, m, H-16),
1,12 (3H, s, H-19), 1,37 (3H, s, H-21), 4,21 (1H, t, J = 7,0 Hz, H-22), 2,05 (2H, m, H23), 1,78 - 1,82 (2H, m, H-24), 1,31 (3H, s, H-26), 1,27 (3H, s, H-27), 0,98 (3H, s, H28), 1,11 (3H, s, H-29), 1,22 (3H, s, H-30), 4,42 (1H, d, J = 7,0 Hz, H-1), 3,49 (1H, m,
H-2), 3,53 (1H, m, H-3), 3,54 (1H, m, H-4), 3,23 - 3,87 (2H, m, H-5), 4,55 (1H, d, J
= 7,5 Hz, H-1), 3,26 (1H, m, H-2), 3,34 (1H, m, H-3), 3,00 (1H, m, H-4), 3,33 (1H,
m, H-5), v 1,28 (3H, d, J = 2 Hz, H-6).

268

C-NMR (125 MHz, MeOD) : 36,4 (C-1), 28,9 (C-2), 90,6 (C-3), 40,8 (C-4),
51,3 (C-5), 23,9 (C-6), 121,0 (C-7), 142,7 (C-8), 148,8 (C-9), 40,3 (C-10), 113,2 (C-11),
29,7 (C-12), 58,9 (C-13), 49,6 (C-14), 34,8 (C-15), 27,7 (C-16), 86,9 (C-17), 178,3 (C18), 23,1 (C-19), 87,1 (C-20), 18,2 (C-21), 81,8 (C-22), 28,5 (C-23), 39,3 (C-24), 82,9
(C-25), 28,9 (C-26), 27,7 (C-27), 17,2 (C-28), 28,6 (C-29), 25,4 (C-30), 106,0 (C-1),
83,1 (C-2), 77,8 (C-3), 71,1 (C-4), 66,4 (C-5), 105,6 (C-1), 76,9 (C-2), 77,5 (C-3),
77,0 (C-4), 73,7 (C-5), v 18,1 (C-6).
13

IV.1.3.7. Hp cht HS7: 2,3,12,21,24R,25- hexahydroxylanosta-8-ene

Tinh th hnh que, mu trng, ph khi lng ESI-MS m/z 490 [M-H2O]+, cng
thc phn t C30H52O6 (M 508).
1
H-NMR (500 Hz, MeOD) : 1,13 (1H, m, Ha-1), 2,04 (1H, m, Hb-1), 3,63 (2H,
m, H-2), 2,93 (1H, dd, J = 1,5, 9,0 Hz, H-3), 1,12 (1H, H-5), 1,54 (1H, m, Ha-6),
2,10(1H, Hb-6), 1,45 (1H, m, Ha-7), 2,21(1H, Hb-7), 2,09(1H, H-11), 4,00 (1H, dd, J =
1,7, 8,5 Hz, H-12), 1,18 (1H, m, Ha-15), 1,76 (1H, m, Hb-15), 1,35 (1H, m, Ha-16), 1,43
(1H, m, Hb-16), 2,41 (1H, m, H-17), 0,65 (3H, s, H-18), 1,04 (3H, s, H-19), 1,38 (1H, m,
H-20), 3,71 (dd, J = 2,5, 12,5 Hz, Ha-21), 3,79 (dd, J = 2,5, 12,5 Hz, Hb-21), 2,10(1H,
H-22), 2,00 (1H, m, Ha-23), 2,75 (1H, m, Hb-23), 3,25 (1H, dd, J = 2,0, 9,5 Hz, H-24),
1,17 (3H, s, H-26), 1,15 1,06 (3H, s, H-30), (3H, s, H-27), 0,82 (3H, s, H-28), v 1,02
(3H, s, H-29).
13
C-NMR (125 MHz, MeOD) : 44,8 (t, C-1), 69,9 (d, C-2), 84,3 (d, C-3), 40,3
(s, C-4), 51,9 (d, C-5), 19,4 (t, C-6), 28,8 (t, C-7), 134,3 (s, C-8), 133,4 (s, C-9), 39,1 (s,
C-10), 27,6 (t, C-11), 74,4 (d, C-12), 51,2 (s, C-13), 51,0 (s, C-14), 32,8 (t, C-15), 29,4
(t, C-16), 39,2 (d, C-17), 17,2 (q, C-18), 20,3 (q, C-19), 44,2 (d, C-20), 61,9 (t, C-21),
28,0 (t, C-22), 33,2 (t, C-23), 79,4 (d, C-24), 73,8 (s, C-25), 25,6 (q, C-26), 25,0 (q, C27), 17,4 (q, C-28), 29,1 (q, C-29), v 24,1 (q, C-30).
IV.1.3.8. Hp cht HS8: 24R-ergosta-7,22-dien-3,5,6-triol.
Tinh th hnh que, mu trng, ph khi lng m/z 412 [M-H2O]+, cng thc phn
t C28H46O3.
1
H-NMR (500 Hz, MeOD) : 1.71 (1H, m, Ha-1), 2.16 (1H, m, Hb-1), 1.97 (1H,
m, Ha-2), 2.26 (1H, m, Hb-2), 4.70 (1H, m, H-3), 2.50 (1H, m, Ha-4), 3.00 (1H, m, Hb-4),
4.28 (1H, br s, H-6), 5.72 (1H, br s, H-7), 2.54 (1H, m, H-9), 1.68 (1H, m, H-11), 1.30
(1H, m, Ha-12), 2.06 (1H, m, Hb-12), 1.92 (1H, m, H-14), 1.54 (2H, H-15), 1.67 (2H, H16), 1.23 (1H, H-17), 0.65 (3H, s, H-18), 1.52 (3H, s, H-19), 2.04 (1H, m, H-20), 1.07
(3H, d, J = 6.5 Hz, H-21), 5.18 (1H, dd, J = 3.0, 15.0 Hz, H-22), 5.22 (1H, dd, J = 2.5,
15.0 Hz, H-23), 1.86 (1H, m, H-24), 1.46 (1H, m, H-25), 0.86 (3H, d, J = 6.5 Hz, H-26),
0.86 (3H, d, J = 6.5 Hz, H-27), v 0.94 (3H, d, J = 6.5 Hz, H-28).
13
C-NMR (125 MHz, MeOD) : 33.9 (t, C-1), 32.6 (t, C-2), 67.5 (d, C-3), 41.9 (t,
C-4), 76.1 (s, C-5), 74.2 (d, C-6), 120.4 (d, C-7), 141.5 (s, C-8), 43.8 (d, C-9), 38.0 (s, C10), 22.4 (t, C-11), 39.9 (t, C-12), 43.8 (s, C-13), 55.2 (d, C-14), 23.4 (t, C-15), 28.4 (t,
C-16), 56.1 (d, C-17), 12.5 (q, C-18), 18.7 (q, C-19), 40.8 (d, C-20), 21.4 (q, C-21), 136.
2 (d, C-22), 132.0 (d, C-23), 43.1 (d, C-24), 33.4 (d, C-25), 20.1 (q, C-26), 19.8 (q, C27), v 17.8 (q, C-28).

269

IV.1.3.9. Hp cht HS9: -Adenosine

Cht tinh th hnh kim mu trng, im chy 234-235oC, ph khi lng ESIMS: m/z 267,8 [M + H]+, 289,8 [M + Na]+ (C10H13N5O4, M = 267).
1
H-NMR (500 MHz, DMSO-d6) H: 8,14 (1H, s, H-2), 8,34 (1H, s, H-8), 5,88
(1H, d, J = 6,5 Hz, H-1), 4,61 (1H, dd, J = 4,0, 10,5, H-2), 4,15 (1H, br s, H-3) 3,97
(1H, dd, J = 3,5, 6,5 Hz, H-4), 3,57 (1H, m, H-5a), 3,68 (1H, m, H-5b), 7,32 (2H, s,
NH2), 5,42 (1H, OH-2), 5,17 (1H, d, J = 3,0, OH-3) v 5,43 (1H, OH-5) ppm
13
C-NMR (125 MHz, DMSO-d6) C: 152,41 (d, C-2), 149,08 (s, C-4), 119,37 (s,
C-5), 156,17 (s, C-6), 139,95 (d, C-8), 87,95 (d, C-1), 73,47 (d, C-2), 70,68 (d, C-3),
85,92 (d, C-4), 61,69 (t, C-5) ppm.

IV.1.4. Xc nh cu trc ha hc ca cc hp cht

IV.1.4.1. Hp cht HS1: Holothurin B
Hp cht HS1 c phn lp t dch chit CHCl3 c dng bt mu trng,
c im nng chy 251-253oC. Cng thc phn t ca n c xc nh l
C41H63O17NaS (M = 882) bng cc kt qu ph khi lng (positive ESI-MS: ti
m/z 905 [M+Na]+) v ph cng hng t ht nhn. Ph IR xut hin cc di pc
c trng ca vng -lactone ti 1720 cm-1, nhm hydroxyl 3400 cm-1, nhm CO-C 1050 cm-1.

Hnh IV.1.4.1.a: Ph 1H-NMR ca HS1

Ph 1H-NMR xut hin cc tn hiu cng hng ca 7 nhm metyl bc ba
(H 0,89, 1,03, 1,12, 1,13, 1,28, 1,50 v 1,52 ppm), mt nhm metyl bc hai ca
mt gc ng xylosepyranosyl [H 1,72 (3H, d, J = 6,0 Hz)], proton olefinic [H
5,40 (1H, d, J = 5,0 Hz)], v cc proton anome ca hai gc ng [H 4,68 (1H, d,
7,8 Hz) v 4,60 (1H, d, J = 7,0 Hz)]. Ph 13C-NMR v cc ph DEPT ca HS1
270

xc nhn s tn ti ca 41 cacbon trong phn t, bao gm 9 cacbon bc bn, 15

nhm metin, 9 nhm metylen v 8 nhm metyl. Nhm cacbonyl c xc nh
bi tn hiu cng hng 174,1, mt ni i b th ba ln >C=CH- [C 154,6
(s)/ 114,2 (d)], 15 cacbon c xc nh l c ni vi nguyn t xy (C 71,3,
72,8, 73,3, 75,6, 75,6, 75,9, 76,0, 80,1, 82,5, 86,0, 89,5, 89,7, 89,8, 104,5 v
104,5 ppm), v ngoi ra cn c mt nhm metylen cng c ni vi xy (C 62,3
ppm).

Hnh IV.1.4.1.b: Ph 13C-NMR ca HS1

Bng IV.1.4.1.: Kt qu ph NMR ca HS1 v Holothurin B
C
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

C ca
Holothurin B
[228]
35,7 (t)
27,4 (t)
88,7 (d)
40,0 (s)
52,8 (d)
20,3 (t)
28,1 (t)
40,9 (d)
153,6 (s)
39,7 (s)
115,5 (d)
71,4 (d)
58,7 (s)
45,9 (s)
27,1 (t)
38,4 (t)

C ca HS1 a,b
36,3 (t)
27,6 (t)
89,5 (d)
40,0 (s)
52,8 (d)
21,1 (t)
27,9 (t)
40,8 (d)
153,7 (s)
39,7 (s)
114,2 (d)
71,3 (d)
59,3 (s)
46,0 (s)
28,1 (t)
38,5 (t)
271

H ca HS1 a,c

3,12 (1H, dd, 10,0, 3,0)

3,40 (1H, m)

5,40 (1H, d, 5,0)

17
18
19
20
21
22
23
24
25
26
27
28
29
30
1
2
3
4
5
1
2
3
4
5
6
a

89,7 (s)
174,4 (s)
18,8 (q)
86,5 (s)
22,5 (q)
80,6 (d)
36,4 (t)
28,7 (t)
81,3 (s)
28,7 (q)
28,1 (q)
21,2 (q)
27,4 (q)
16,7 (q)
105,7 (d)
83,4 (d)
76,6 (d)
75,1 (d)
64,0 (t)
105,2 (d)
76,0 (d)
77,4 (d)
76,6 (d)
73,4 (d)
18,5 (q)

89,7 (s)
174,9 (s)
18,4 (q)
86,0 (s)
22,5 (q)
80,1 (d)
36,8 (t)
28,7 (t)
82,5 (s)
28,7 (q)
28,1 (q)
21,0 (q)
27,4 (q)
16,7 (q)
104,5 (d)
82,2 (d)
75,6 (d)
73,3 (d)
62,9 (t)
104,5 (d)
75,6 (d)
76,0 (d)
75,9 (d)
72,8 (d)
17,8 (q)

1,13 (3H, s)
1,52 (3H, s)

1,12 (3H, s)
1,03 (3H, s)
0,89 (3H, s)
1,28 (3H, s)
1,50 (3H, s)
4,60 (1H, d, 7,0)

4,68 (1H, d, 7,8)

1,72 (3H, d, 6,0)

o trong CDCl3+CD3OD+D2O; b125 MHz; c500 MHz.

Hnh IV.1.4.1.c: Ph DEPT 90, DEPT 135 v 13C-NMR ca HS1

Cc gi tr ph ca HS1 c so snh vi cc gi tr ph c cng b
ca holothurin A, B, C v D, cc hp cht c kiu khung c trng c phn lp
272

t loi Holothuria forskalli [227[219]228]. Kt qu so snh cho thy rng tt c

cc gi tr ph ca HS1 u tng ng ph hp vi cc kt qu ph ca
holothurin B (Bng IV.1.4.1). T cc d kin ph nu trn, HS1 c nhn dng
l
holothurin
B:
3-O-(2-O--D-quinovopyranosyl--D-xylopyranosyl)holothurigenol-4-O-sodium sulfate, hp cht ny cng c phn lp t
ccloi: Holothuria leucospilota, H. lubrica , H. floridiana, H. edulis, H. atra,
Stichopus chloronotus, Actinopyga flammea, Actinopyga agassizi v Thelenota
ananas. Tuy nhin y cng l ln u, holothurin B c phn lp t Holothuria
scabra sinh sng ti vng bin Vit Nam.
21

O
HO

11

4
5'

16
15

28

30

OH

25

OH

14

10

22

13
9

26

18
17

24

20
12

19

23

29

1'
2'

N a O 3S O

6"

CH3
O

5"

OH

OH

1"
2"

OH

Hnh IV.1.4.1.d: Cu trc ho hc ca HS1

Hnh IV.1.4.1.e: Ph khi lng ESI-MS ca HS1

273

27

IV.1.4.2. Hp cht HS2: Holothurin A

Hp cht HS2 c tch t dch chit CHCl3, c dng bt mu trng, nhit
nng chy 228-230oC. Ph IR xut hin cc di hp th 3426 cm-1 (br) v
1068 cm-1 rt in hnh cho cc hp cht c nhnh ng. Ngoi ra cng trn ph
IR cn xut hin nh hp th 1763 cm-1 ca 1 vng nm lacton.

Hnh IV.1.4.2.a: Ph hng ngoi ca HS2

Hnh IV.1.4.2.b: Ph 1H-NMR ca HS2

274

Ph 1H-NMR ca HS2 c cc tn hiu singlet ca 7 nhm methyl bc 3

vi H 0,81; 1,00; 1,04; 1,16; 1,19; 1,21; 1,37 ppm v mt nhm methyl bc 2 vi
tn hiu doublet ti H 1,24 ppm. Trong phn t ny cn c 1 nhm metoxy H
3,48 (3H, s) v 1 lin kt dng >C=CH- vi H 5,24 (1H, d, J = 4,5 Hz). T ph
ny cn quan st c s c mt ca 4 gc ng trong phn t vi cc tn hiu
doublet c trng ca cc proton anomeric H 4.35 (1H, d, J = 8.0 Hz); 4.49 (1H,
d, J = 8.0 Hz); 4.38 (1H, J = 8.0 Hz); v 4.33 (1H, d, J = 7.5 Hz). vng trng
yu cn c tn hiu cng hng ca mt s proton thuc cc nhm hydroxy.

Hnh IV.1.4.2.c: Ph 13C-NMR v cc ph DEPT ca HS2

275

Ph 13C-NMR v cc ph DEPT cho thy trong phn t HS2 c 54 cacbon,

bao gm 9 nhm CH3, 11 nhm CH2, 25 nhm CH, 8 cacbon bc bn v 1 nhm
cacbonyl vi C 173,5 ppm. Cc du hiu ca ni i [C 114,5(d)/152,6(s)],
nhm metoxy C 59,8 ppm v 4 cacbon anomeric ca cc gc ng C 102,9;
103,6; 103,8; 103,9 ppm cng mt ln na c khng nh. Cng trn cc ph
ny trong vng t 60 n 90 ppm l cc tn hiu cng hng ca 25 cacbon lin
kt trc tip vi oxi (ch yu ca cc gc ng), bao gm 3 nhm CH2, 19 nhm
CH v 3 cacbon bc bn.

Hnh IV.1.4.2.d: Ph HSQC ca HS2

Bng IV.1.4.2.a: Kt qu ph NMR phn aglycon ca HS2 v Holothurin A

1
2
3
4
5
6

Ca, c
[228]
35,5
27,4
88,7
40,0
52,9
20,3

7
8
9
10

28,2
41,0
153,7
39,7

Cb, c
35,6
26,2
87,6
39,2
51,9
20,4
27,3
39,9
152,6
38,8

Hb, d
1,67 -1,75 (2H, m)
1,62 -1,84 (2H, m)
3,39 (1H, m)
0,85 (1H, m)
1,44 (1H, d, 11,5 Hz)
1,63 (1H, m)
1,32 -1,68 (2H, m)
2,88 (1H, dd, J = 5,0 Hz)

276

HMBC

C-1 Xyl
C-1; C-7; C-29

C-9; C-14

11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
OH-17
OH-12
a

115,6
71,7
59,0
46,0
27,2
38,6
89,7
174,3
19,0
86,5
22,5
80,7
36,6
28,7
81,3
28,7
28,2
16,7
27,4
21,4

114,5
70,2
57,4
44,8
35,8
34,4
88,4
173,5
21,8
86,0
18,4
79,4
27,2
37,8
80,8
28,4
27,1
16,1
27,4
19,4

5,24 (1H, d, J = 4,5 Hz)

4,41 (1H, m)

C-10; C-13; C-8

1,00- 1,38 (2H, m)

1,85 -2,65 (2H, m)

C-13; C-14; C-20

1,04 (3H, s)

C-1, C-5, C-9, C-10

1,37 (3H, s)
4,10 (2H, t, 7,0 Hz)
1,90- 2,00 (2H, m)
1,63 - 1,73 (2H, m)

C-17, C-20, C-22

C-17; C-20, C-21
C-20; C-24; C-25
C-23; C-25, C-26, C-27

1,21 (3H, s)
1,16 (3H, s)
0,81 (3H, s)
1,00 (3H, s)
1,19 (3H, s)
6,50 (s)
6,12 (d, 3,25 Hz)

C-24, C-25, C-27

C-24, C-25, C-26
C-3, C-4, C-5, C-29
C-3, C-4, C-5, C-28
C-8, C-14, C-15, C-13
C-13, C-16, C-17
C-13

C5D5N; bDMSO; c125MHz; d500MHz.

Hnh IV.1.4.2.e: Ph khi lng ESI-MS ca hp cht HS2

277

21

O
HO

19

Xyl
4'

27

OH

15

6
29

28

2'

Qui
4''
6'''

25

1'

NaO3SO
6'' CH3

26

30

O O

OH

17
13

3
5'

22
20

12

11

10

O
18

24

23

O O
1''

OH

CH2OH

4'''

6''''

CH2OH
O

OH

MeGlc
4''''
OH

1''''

OCH3

Glc

O O

2''

OH

1'''
2'''

OH

2''''

OH

Hnh IV.1.4.2.f: Cu trc ho hc ca HS2

T nhng thng tin trn c th nhn dng HS2 l mt triterpene glycoside
4 ng. Sau khi ghp chuyn dch ho hc ca C vi H cho tt c cc v tr
theo ph HSQC chng ti nhn thy d liu ph NMR ca HS2 hon ton trng
vi holothurin A [228]. Cc kt qu c trnh by trong bng IV.1.4.2.a v b.
Kt qu ph khi lng ESI-MS ca HS2 cho cc pic ion phn t m/z
1243,3 [M+Na]+ v m/z 1197,4 [M-Na]- hon ton ph hp vi cng thc phn t
C54H85O27SNa (M=1220) ca holothurin A.
Bng IV.1.4.2.b: Kt qu ph NMR phn chui ng ca HS2 v Holothurin A
Ca, c
[228]

Cb, c
HS2

Xyl 1
2
3
4
5
Qui 1
2
3
4

105,7
83,1
76,7
74,9
63,8
105,2
76,0
76,7
86,5

103,8
81,4
74,8
74,4
63,0
103,6
74,5
74,3
86,2

4,35 (1H, d, J = 8,0 Hz)

3,36 (1H, m)
3,05 (1H, dd, 3,5; 3,5 Hz)
3,95 (1H, m)
3,23 - 3,95 (2H, m)
4,49 (1H, d, J = 8,0 Hz)
3,51 (1H, m)
3,30 (1H, m)
2,99 (1H, m)

5
6
Glc 1
2
3

71,7
17,9
105,1
73,9
88,1

70,1
17,3
102,9
72,1
87,9

3,33 (1H, m)
1,24 (3H, s)
4,38 (1H, J = 8,0 Hz)
3,21 (1H, m)
3,04 (1H, m)

Hb, d
HS2

278

HMBC
C-3; C-5 Xyl
C1 Xyl; C-3 Xyl
C-5 Xyl
C-1 Xyl, C-4 Xyl
C-2 Xyl
C-3 Qui
C-4 Qui
C-3 Qui, C-5 Qui
C-6 Qui ; C-1 Glc
C-4 Qui, C-5 Qui
C-4 Qui

4
5
6
OMeGlc
1
2
3
4
5
6
OCH3
OH-3
OH-3
OH-4
OH-6
OH-2
OH-4
OH-6
a

70,8
77,3
62,2

68,4
76,1
60,7

3,22 (1H, m)
3,31 (1H, m)
3,41 - 3,68 (2H, m)

C-5 Glc
C-4 Glc; C-6 Glc
C-5 Glc

104,6
74,9
87,7

103,9
73,5
85,9

4,33 (1H, d, J = 7,5 Hz)

3,15 (1H, m)
2,98 (1H, m)

69,5
78,0
61,9
60,5

69,3
76,7
60,9
59,8

3,13 (1H, m)
3,21 (1H, m)
3,41 - 3,68 (2H, m)
3,48 (3H, s)
5,80 (d, 2,3 Hz)
4,57 (d, 2,0 Hz)
4,67 (d, 3,0 Hz)
4,64 (t, 5,5 Hz)
5,08 (d, 3,5 Hz)
5,12 (d, 6,0 Hz)
4,59 (t, 5,5 Hz)

C-3 Glc
C-1 MeGlc
C-2 MeGlc, OCH3
C-4 MeGlc
C-6, C-5 MeGlc
C-4 MeGlc
C-3 MeGlc
C-2, C-4 Xyl
C-2, C-3 Qui
C-4, C-5 Glc
C-3 MeGlc
C-3, C-4, C-5 MeGlc
C-5, C-6 MeGlc

C5D5N; bDMSO; c125MHz; d500MHz.

Trn ph cng hng t hai chiu HMBC cc tng tc xa gia H-3 vi C1; H-1; H-1 vi C-2; H4 vi C-1 v H-1 vi C-4; H-1 vi C-3
chng t chui ng lin kt vi v tr C-3 ca khung holosthane theo trt t
MeGlc(1-3)-Glc(1-4)-Qui(1-2)-Xyl(1-3)-Aglycon. Cng t ph ny, tn hiu ca
cc nhm OH v tr C-17, C-12 trn khung holosthane v ca cc gc ng
cng c xc nh. Ngoi ra trt t lin kt trong ton b phn t HS2 cn c
khng nh bng cc tng tc xa khc v c ch ra trong bng IV.1.4.2 a-b v
hnh IV.1.4.2.h. Mt khc, khng nh r cu trc ca cc gc ng ca HS2
chng ti s dng phng php thu phn kt hp vi sc k lp mng TLC.
Kt qu so snh cho thy cc cu t ng ca HS2 c gi tr Rf v mu sc hon
ton ph hp vi cc ng chun -D-xylopyranose (X), -D-quinovopyranose
(Q), -D-Glucopyranose (G) v -D-3-O-methylglucopyranose (MG), (hnh
4.10). T nhng kt qu phn tch nu trn, hp cht HS2 c nhn dng l
holothurin A, vi tn khoa hc l 3-O-[-D-3-O-methylglucopyranosyl-(1-3)-D-glucopyranosyl-(1-4)--D-quinovopyranosyl-(1-2)--D-xylopyranosyl]holothurigenol-4-O-natri sulfat.

279

Hnh IV.1.4.2.g: Ph HMBC ca HS2

O
HO

19

17

1
9

4'

30
6

1'

28

29

O
4''

15

2'

NaO3SO
6'' CH3

6'''

OH

O O

OH

16
10

5'

22
20

13

26
25

O
18

12

11

24

23

21

1''

OH

CH2OH
O O

2''

OH
6''''

4'''

CH2OH
O

4''''

OH

1'''
2'''

OH

OCH3

OH

1''''
2''''

OH

Hnh IV.1.4.2.h: Mt s tng tc HMBC chnh trong phn t HS2.

280

27

X: ng chun D-xylopyranose
Q: ng chun D-quinovopyranose
G: ng chun D-Glucopyranose
MG: ng chun D-3-O-methylglucopyranose
T: Mu hn hp ng ca HS2

Hnh IV.1.4.2.i: So snh hn hp ng ca HS2 vi cc ng chun trn TLC

IV.1.4.3. Hp cht HS3: Holothurin A2
Hp cht HS3 tch c t dch chit butanol, c dng bt mu trng, c
im chy 246-2480 C v []D20: -11,050 (C=0,26; C2H5OH). Cng thc phn t
ca n c xc nh l C54H87O26SNa bng cc kt qu ph khi lng (positive
ESI-MS ti m/z 1229,6 [M+Na]+) v ph cng hng t ht nhn.

Hnh IV.1.4.3.a: Ph 1H-NMR ca hp cht HS3

Ph 1H-NMR cho thy s xut hin ca 1 nhm metoxy v tr 3,49
(3H, s), cc tn hiu n ca 5 nhm metyl bc 3 0,81, 1,01, 1,03, 1,20, 1,42
ppm. Ph ny cng xut hin tn hiu vch kp v tr 0,86 vi hng s tng
tc J = 6,5 Hz v tch phn l 6H chng t s c mt ca 2 nhm metyl bc 2,
mt nhm metyl bc 2 khc vi tn hiu doublet v tr H 1,24 (3H, d, J = 6
281

Hz). Bn cnh , cc tn hiu trn ph ny cng th hin s c mt ca 1 ni i

ni vng th 3 ln v tr 5.25 (1H, d, J = 4 Hz). S c mt ca proton nhm
CH gn oxy cng c khng nh trn ph proton vi 4.43 (1H, d, J = 4.0 Hz).
Ngoi ra, s c mt ca 4 gc ng trong phn t cng c ghi nhn, cc
proton anomeric vng trng yu vi J c trng (t 7 n 8 Hz): 4,32 (1H, d,
J = 7,0 Hz), 4,49 (1H, d, J = 8,0 Hz), 4,38 (1H, d, J = 8,0 Hz), v 4,34 (1H, d, J
= 8,0 Hz).
21

O
HO

19

10

4'

OH

6'' CH3

4''
6'''

27

15

6
28

29

2'
O O
1''

OH

CH2OH
O O

6''''

CH2 OH
O

O
4''''

OCH3
OH

1''''

2''

OH

4'''
1'''

OH

2'''

OH

2''''

OH

Hnh IV.1.4.3.b. Cu trc ha hc ca HS3

Hnh IV.1.4.3.c. Ph 13C-NMR ca HS3

282

26
25

23

1'

24

OH

30

O O

NaO3SO

17
13

3
5'

20

18
12

11

22

Hnh IV.1.4.3.d. Ph 13C-NMR v cc ph DEPT ca HS3

Ph 13C-NMR v DEPT ca HS3 cho thy s c mt ca 54C trong phn
t vi 9 nhm CH3; 12 nhm CH2; 25 nhm CH v 8C bc 4. Trong c 1 nhm
carbonyl vi 173,6 ppm v 1 lin kt kiu >C=CH [114,5 (d)/ 152,6 (s)].
C 1 nhm CH3 cng hng vng trng yu 59,2, y l 1 nhm metoxy ca
1 gc ng. Trong 12 nhm CH2, c 3 nhm lin kt trc tip vi oxi, cng
hng ti 62,98, 60,85, 60,74 ppm. Trong cc nguyn t C lin kt vi oxi cn
c 21 nhm CH trong tng s 25 nhm cng hng vng trng yu (C 68
n 88 ppm) v 2 cacbon bc 4 [ 88,13 v 86,2]. Cng trn ph ny, 4 cacbon
anomeric cng c xc nh vng trng yu: 102,92, 103,60, 103,83 v
103,9 khng nh r s c mt ca 4 gc ng trong phn t.
xc nh cc mi lin kt trc tip C-H, chng ti tin hnh o ph
HSQC, kt qu cho thy hp cht HS3 kh ging vi holothurin A, c bit l
chui lin kt ca 4 gc ng vi s c mt ca 1 nhm NaSO3-. Phn khung
Holosthan ca 2 phn t ny cng ging nhau tt c cc v tr t C-1 n C-21 v
t C-28 n C-30, nhng so vi holothurin A th HS3 khng cn cc tn hiu C
81,0 v 81,4 ppm ca nhm ete vng thuc phn nhnh bn t C-22 n C-27,
m thay vo l cc tn hiu vng trng mnh ca 1 nhnh isohexan.

283

Hnh IV.1.4.3.e. Ph HSQC ca hp cht HS3

Vi nhng thng tin trn kt hp vic so snh vi cc ti liu thu thp
c, chng ti nhn thy cc d liu ph ca HS3 hon ton trng vi
holothurin A2 - mt hp cht tch c t loi Holothuria floridana.
Bng IV.1.4.3: Kt qu ph NMR ca HS3 v Holothurin A2
C
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

Ca, c
[229]
36,6
27,1
88,6
40,0
52,7
21,2
28,0
40,0
153,9
39,7
115,5
71,4
58,6
46,4
38,9
36,0

Cb, c
HS3
35,8
26,1
87,5
38,9
51,9
20,3
27,1
40,0
152,6
39,8
114,5
70,1
57,4
45,3
35,6
34,7

Hb, d
HS3
1,37 - 1,75 (2H, m)
1,64 - 1,87 (2H, m)
3,39 (1H, m)

HMBC

C-1 Xyl

0,88 (1H, m)
1,67 (2H, m)
1,35 - 1,67 (2H, m)
2,88 (1H, dd, 4,5; 5,0 Hz)

5,26 (1H, d, 4,0 Hz)

4,43 (1H, d, 5,0 Hz)

1,05 - 1,70 (2H, m)

1,95 - 2,41 (2H, m)

284

C-13; C-8

17
18
19

89,3
174,8
22,5

88,13
173,6
21,9

1,03 (3H, s)

C-1, C-5, C-9, C10

20
21
22
23
24
25
26
27
28

87,2
23,0
36,6
22,2
39,7
28,0
22,6
22,6
16,7

86,2
22,4
37,8
21,3
38,9
27,2
22,5
22,5
16,2

1,43 (3H, s)
1,65 (2H, m)
1,31 - 1,40
1,18 (2H, m)
1,55 (m)
0,87 (3H, s)
0,86 (3H, s)
0,81 (3H, s)

C-17, C-20, C-22

C-20, C-23

29

28,0

27,4

1,02 (3H, s)

30

20,1

19,2

1,20 (3H, s)

Xyl 1
2
3
4
5
Qui 1
2
3
4

105,1
83,0
76,2
75,4
63,9
105,1
76,2
75,6
87,6

103,83
81,4
74,8
74,2
62,98
103,60
74,5
74,2
86,1

4,32 (1H, d, 7,0 Hz)

3,37 (1H, m)
3,08 (1H, m)
3,93 (1H, m)
3,21 - 3,95 (2H, m)
4,49 (1H, d, 8,0 Hz)
3,52 (1H, m)
3,29 (1H, m)
3,00 (1H, m)

5
6
Glc 1
2
3
4
5
6
OMeGlc
1
2
3
4
5
6
OCH3

71,7
18,1
104,6
73,9
88,2
69,9
77,7
62,4

70,10
17,3
102,92
72,1
87,9
68,4
76,07
60,74

3,33 (1H, m)
1,25 (3H, s)
4,38 (1H, d, 8,0 Hz)
3,21 (1H, m)
3,03 (1H, m)
3,21 (1H, m)
3,31 (1H, m)
3,41 - 3,68 (2H, m)

105,4
74,9
87,7
70,8
78,0
62,4
60,4

103,9
73,5
85,9
69,26
76,9
60,85
59,82

4,34 (1H, d, 8,0 Hz)

3,13 (1H, m)
3,00 (1H, m)
3,14 (1H, m)
3,22 (1H, m)
3,41 - 3,68 (2H, m)
3,49 (3H, s)

C5D5N; bDMSO; c125MHz; d500MHz.

285

C-25, C-26, C-27

C-26, C-27
C-24, C-25, C-27
C-24, C-25, C-26
C-3, C-4, C-5, C29
C-3, C-4, C-5, C28
C-8, C-14, C-15,
C-13
C-3
C1 Xyl
C-1 Xyl, C-4 Xyl
C-1 Xyl, C-4 Xyl
C-2 Xyl
C-3 Qui
C-2 Qui
C-3, C-5,
C-6 Qui, C-1 Glc
C-4 Qui, C-5 Qui
C-4 Qui
C-1, C-3 Glc
C-5 Glc

C-3 Glc
C-1 MeGlc
C-2 MeGlc, OCH3
C-3, C-5 MeGlc

C-3 MeGlc

Hnh IV.1.4.3.f. Ph HMBC ca hp cht HS3

21

O
HO

19

10

4'

OH

4''

15

6
28

29

O
1''

OH

O O

O
1''''

OCH3
OH

27

CH2OH

CH2OH

4''''

25

23

OH

1'

6'''

26

2'

6'' CH3

6''''

24

30

O O

NaO3SO

13
17

3
5'

20

18
12

11

22

2''''

OH

2''

OH

4'''
1'''

OH

2'''

OH

Hnh IV.1.4.3.g. Mt s tng tc chnh trong phn t HS3

HS3 l mt phn t ln, do xc minh r cc v tr trong phn t
chng ti tin hnh o ph cng hng t 2 chiu HMBC. Kt qu cho thy
v tr C-21 c cc tng tc xa: H-21 vi C-17; C-20 v C-22, ngc li H-22

286

cng tng tc vi C-20; C-23. cc v tr tip theo thuc nhnh bn cng xut
hin cc tng tc H-24 vi C-25; C-26; C-27; H-25 vi C-26; C-27; H-26 v H27 cng c tng tc vi C-25; C-24, C-26 v C-27. Cc bng chng ny
khng nh cc v tr t C-22 n C-27 l 1 nhnh isohexan v c gn vo
khung holasthane v tr C-20.

Hnh IV.1.4.3.h. Ph khi lng ESI-MS ca hp cht HS3

Lin kt glycozit gia khung holasthane vi chui ng ti v tr C-3
cng c xc nhn thng qua cc tng tc HMBC ca H-3 vi C-1 xylo v H1 xylo vi C-3. Ngoi ra cn quan st c cc tng tc H-1 quino vi C-1
xylo; H-4 quino vi C-1 gluco v H-1 gluco vi C-4 quino; H-1
OMeGluco vi C-3 gluco. Nhng tng tc ny chng minh trt t lin kt
trong chui ng l Xylo-Quino-Gluco-OMeGluco. Nhng tng tc khc
trong phn t c trnh by trong bng IV.1.4.3 v hnh IV.1.4.3.g. T cc kt
qu nu trn, hp cht HS3 c nhn dng l holothurin A2 vi tn khoa hc l
3,12,17
-trihydroxyholost-9(11)-ene
3-O-[(3-O-methyl)--Dglucopyralnosyl-(13)--D-glucopyranosyl-(14)--D-quinovopyranosyl(12)-(4-O-sunfo)--D-xylopyranoside]natri.
IV.1.4.4. Hp cht HS4: Holothurin A3 (Cht mi)
Hp cht HS4 c phn lp t pha nc, c dng bt mu trng. Cng
thc phn t ca HS4 c xc nh l C54H85O28SNa thng qua kt qu ph khi
lng ESI-MS ti m/z 1259,3 [M + Na]+ (positive) v 1235,5 [M - H]+ v ph
khi lng phn gii cao HR FAB-MS: m/z 1259,4900 [M + Na]+ (positive) v
kt qu tnh phn tch nguyn t cho cng thc C54H85O28SNa2 l 1259,4743.

287

Hnh IV.1.4.4.a: Ph 1H-NMR ca HS4

O
O

HO
12

17

OH
OH

O O
OH

NaO3SO
CH3
O

OH
CH2OH

CH2OH

O O

OH
O
OCH3
OH

OH
OH

OH

Hnh IV.1.4.4.b: Cu trc ha hc ca HS4

Trn ph 1HNMR, vng trng mnh xut hin 7 tn hiu singlet ca 7
nhm metyl bc 3 (H 0,81; 1,02; 1,05; 1,071; 1,079; 1,18; 1,56 ppm) v tn hiu
doublet ca 1 nhm metyl bc 2 vi H 1,24 (3H, d, J = 6,0 Hz). Ngoi ra trong
phn t cn tn ti mt nhm metoxy cng hng ti H 3,49 (3H, s). Cng trn
ph ny, vng trng yu cn quan st c 1 proton olefin ca 1 ni i b th
3 ln (H 5,26 ppm) v cc tn hiu doublet ca 4 proton anome ca 4 n v
ng ti : H 4,32 (1H, d, J = 7,5 Hz), 4,34 (1H, d, J = 7,5 Hz), 4,38 (1H, d, 8,0
Hz), v 4,49 (1H, d, J = 7,5 Hz).

288

Hnh IV.1.4.4.c: Ph 13C-NMR ca HS4

Hnh IV.1.4.4.d: Ph 13C-NMR v cc ph DEPT ca HS4

Ph 13C-NMR v DEPT 135 ca HS4 cho thy s c mt ca 54 nguyn t
cacbon trong phn t, bao gm 9 nhm metyl, 11 nhm metylen, 24 nhm metin
v 10 cacbon bc 4. Trong c 2 nhm cacbonyl vi tn hiu cng hng ti
172,6 ppm v 208,1ppm v 1 ni i b th 3 v tr [ 114,5(d)/152,8 (s)]. Bn
nhm mtin vi tn hiu cng hng ti 103,92, 103,86, 103,62, 102,95 c
xc nh l cc cacbon anme ca 4 gc ng. Ngoi ra cn c 24 cacbon khc
cng lin kt trc tip vi xi, bao gm: 17 nhm metin ( 87,5, 69,5, 81,4, 74,8,

289

74,2, 74,5, 74,2, 86,2, 70,1, 72,1, 87,9, 68,4, 76,1, 73,5, 85,9, 69,27, 76,8), 3
cacbon bc 4 ( 91,3, 86,2, 68,0 ppm), 3 nhm metylen ( 63,0, 60,9, 60,7) v 1
nhm metoxy ( 59,8).

Hnh IV.1.4.4.e: Ph HSQC ca HS4

Gi tr chuyn dch ha hc ca cc cacbon c gn vi cc proton
tng ng thng qua ph HSQC. Kt qu chi tit c tng hp trong bng
IV.1.4.4. Cng thc phn t C54H85O28SNa ca HS4 cho php xc nh hp cht
ny c khng no l 12, trong khi cc ph NMR khng nh ch tn ti 3 lin
kt (2 nhm cacbonyl v 1 ni di kiu b th 3 v tr), iu ny chng t trong
phn t c 9 cu trc vng, trong c 4 vng pyranosyl ca 4 gc ng.
Vi cc thng tin thu c, HS4 c nhn nh c cu trc kiu
tritecpen-glyccsit, bao gm khung holasthane lin kt vi chui 4 phn t ng
(xyloz- Quinovz- Glucoz-OMe Glucz). Kt qu ph 13C-NMR v DEPT
ca HS4 v holothurin A2 hon ton trng khp phn chui 4 ng v cc v tr
t C-1 n C-21, t C-28 n C-30 ca khung holasthane. S khc bit gia 2
phn t ny l nhnh bn t v tr C-22 n C-27, c th so vi holothurin A2 th
HS4 c thm 1 nhm cacbonyl v 1 nhm ancol bc 3.
Da vo cc tng tc xa trn ph HMBC, cc v tr ca nhnh bn t C-22
n C-27 c xc nh. H-21 ngoi cc tng tc vi C-20 v C-17, cn
tng tc vi C-22 (C 208,1 ppm), iu ny khng nh v tr 22 l nhm
cacbonyl. Bn cnh l tng tc ca H-23 vi C-22; H-24 vi C-23; C-25; C26 v C-27. C-25 l cacbon bc 4 lin kt vi 2 nhm CH3 (v tr 26 v 27) v 1
nhm OH. Ngoi vic tng tc qua li vi nhau, proton ti cc v tr 26, 27 cn

290

tng tc vi C-25 v C-24. c bit proton ca OH-25 (H 4,23,s) cng th hin

tng tc vi C-26; C-27; C-25 v C-24. Hn na cc gi tr ph ca nhnh ny
cng hon ton trng khp vi phn nhnh 22-one-25-hydroxyl isohexan trong
ti liu [230]

Hnh IV.1.4.4.f. Ph HMBC ca HS4

21

O
24

18

O
22

HO

4'

OH

15

6
1'

NaO3SO

17

30

O O

OH

13

26

5'

OH
12

19

27
25

29

28

2'

CH3
O O
4''

6'''

1''

OH

CH2OH
6''''
CH2OH

O O

2''
OH

4'''

1'''

O
OH

4''''

OCH3

1''''

OH

OH
3''''
OH

Hnh IV.1.4.4.g. Mt s tng tc HMBC chnh trong phn t HS4

291

Bng IV.1.4.4. Kt qu ph NMR ca HS3 v HS4

STT
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
OH-17
OH-25
Xyl 1
2
3
4
5
Qui 1
2
3

Ca, b
HS3
35,8
26,1
87,5
38,9
51,9
20,3
27,1
40,0
152,6
39,8
114,5
70,1
57,4
45,3
35,6
34,7
88,13
173,6
21,9
86,2
22,4
216,1*
32,7*
38,4*
69,1*
29,8*
30,0*
16,2
27,4
19,2

Ca, b
HS4
35,7
26,2
87,5
38,9
51,9
20,6
27,4
40,0
152,8
39,7
114,5
69,5
57,3
45,2
37,8
35,8
91,3
172,6
21,9
86,2
20,6
208,1
33,5
36,4
68,0
29,1
29,4
16,1
27,4
19,0

103,83
81,4
74,8
74,2
62,98
103,60
74,5
74,2

103,86
81,4
74,8
74,2
63,0
103,62
74,5
74,2

Ha, c
HS4
1,41 - 1,74 (2H, m)
1,63 - 1,85 (2H, m)
3,34 (1H, m)

HMBC

C-1 Xyl

0,88 (1H, m)
1,67 (2H, m)
1,35 - 1,67 (2H, m)
2,87 (1H, m)

C-10

5,26 (1H, d, 4,5 Hz)

4,48 (1H, m)

C-13; C-8

1,69 2,05 (2H, m)

1,70 (2H, m)

C-14

1,05 (3H, s)

C-5, C-9, C-10

1,56 (3H, s)

C-17, C-20, C-22

2,64 (2H, m)
1,54 - 1,61 (2H, m)

C-22
C-23, C-25, C-26, C-27

1,07 (3H, s)
1,07 (3H, s)
0,81 (3H, s)
1,02 (3H, s)
1,18 (3H, s)
7,03 (1H, s)
4,23 (1H, s)
4,32 (1H, d, 7,5 Hz)
3,34 (1H, m)
3,07 (1H, m)
3,94 (1H, m)
3,20 - 3,93 (2H, m)
4,49 (1H, d, 7,5 Hz)
3,51 (1H, m)
3,29 (1H, m)

C-24, C-25, C-27

C-24, C-25, C-26
C-3, C-4, C-5, C-29
C-3, C-4, C-5, C-28
C-8, C-14, C-15, C-13
C-13, C-15, C-17
C-24, C-25, C-26, C-27
C-3
C-1 Xyl
C-1 Xyl

292

C-1 Xyl, C-4 Xyl

C-2 Xyl
C-3 Qui
C-2 Qui

86,1

86,2

3,00 (1H, m)

5
6
Glc 1
2
3
4
5
6
OMeGlc
1
2
3
4
5
6
OCH3
OH-3

70,10
17,3
102,92
72,1
87,9
68,4
76,07
60,74

70,1
17,3
102,95
72,1
87,9
68,4
76,1
60,76

3,33 (1H, m)
1,25 (3H, s)
4,38 (1H, d, 8,0 Hz)
3,21 (1H, m)
3,03 (1H, m)
3,21 (1H, m)
3,31 (1H, m)
3,41 - 3,68 (2H, m)

103,9
73,5
85,9
69,26
76,9
60,85
59,82

103,92
73,5
85,9
69,27
76,8
60,86
59,87

4,34 (1H, d, 7,5 Hz)

3,15 (1H, m)
2,99 (1H, m)
3,14 (1H, m)
3,21 (1H, m)
3,41 - 3,68 (2H, m)
3,49 (3H, s)
5,80 (1H, s)

C-3, C-5, C-6 Qui,

C-1 Glc
C-4 Qui, C-5 Qui
C-4 Qui

C-4 Glc
C-3 Glc
C-1 OMeGlc
C-2 OMeGlc, OCH3
C-5 OMeGlc

C-3 OMeGlc
C-2 Xyl, C-3 Xyl

*phn nhnh 22-on-25-hydroxyl isohexan trong ti liu [230], aDMSO; b125MHz; c500MHz

Trn khung holosthan, cc tng tc gia H-5 vi C-10; gia H-11 vi C13; C-8; gia H-15 vi C-14; gia H-19 vi C-5, C-9, C-10; gia H-28 vi C-3,
C-4, C-5, C-29; gia H-29 vi C-3, C-4, C-5, C-28; gia H-30 vi C-8, C-14, C15, C-13 cng c ghi nhn. ng ch l OH-17 (H 7,03, s) cng th hin
tng tc vi C-13, C-15 v C-17.

Hnh IV.1.4.4.h. Ph khi lng ESI-MS ca HS4

293

 chui ng, th t lin kt MeGlc(1-3)-Glc(1-4)-Qui(1-2)-Xyl(1-3)Aglycon cng c xc nhn. Trc tin l tng tc xa H-3 vi C1xyl; H-1xyl
vi C-3, khng nh khung holosthan gn trc tip vi ng xylose. Tip theo l
tng tc gia prton anomeric H-1 Quino vi C-2xylo; H-1Gluco vi C4Quino; gia H-4Quino vi C-1Gluco v H-1OMeGluco vi C3Gluco. Trt t cc v tr trong tng gc ng cng c lm sng t nh cc
tng tc xa. gc OMeGluco xut hin cc tng tc ca: H-OMe vi C-3;
H-4 vi C-5; H-3 vi C-OMe v C-2, H-2 vi C-1, cn gc
Gluco l tng tc: H-6vi C-4. ng Quino c tng tc H-6 vi C-5
v C4; H-4 vi C-3, C-5, C-6; H-3 vi C-2; H-2 vi C-3 v H-1
vi C-2. Cui cng l ng Xylo c cc tng tc H-5 vi C-1, C-4; H-3
vi C-1 v H-2 vi C-1. V tr nhm OH-3 ca ng xyloz cng c xc
nhn bng cc tng tc OH-3 (5.80 ppm) vi C-2 v C-3.

Hnh IV.1.4.4.i. Ph khi lng phn gii cao HR-ESI-MS ca HS4

T cc kt qu ph khi lng EI-MS, HR-ESI-MS, ph 1D, 2D NMR nu
trn, hp cht HS4 c xc nh l 3,12,17,25 -Tetrahydroxyholost-9(11)ene-22-one 3-O-[(3-O-methyl)--D-glucopyralnosyl-(13)--D-glucopyranosyl-(14)--D-quinovopyranosyl-(12)-(4-O-sunfo)--D-xylopyranoside]natri.
y l mt hp cht mi, ln u tin phn lp c t thin nhin v c
chng ti t tn l holothurin A3
IV.1.4.5. Hp cht HS5: Holothurin A4
Hp cht HS5 tch c t dch chit pha nc c dng bt trng. Cng
thc phn t C54H87O27SNa (M=1222) ca HS5 c xc nh bng kt qu ph
khi lng ESI-MS (positive cho pic ion phn t m/z 1245,2 [M+Na]+ v negative
cho pic ion m/z 1221,4 [M-H]-) v ph cng hng t ht nhn.

294

Hnh IV.1.4.5.a: Ph 1H-NMR ca hp cht HS5

Trn ph 1H-NMR vng trng mnh 5 tn hiu singlet ca 5 nhm
metyl bc 3 (H 0,81; 1,00; 1,03; 1,20; 1,42 ppm) v 3 nhm metyl bc 2 (H
0,83; 0,84; 1,25 ppm). Trong phn t HS5 cn mt nhm metoxy cho tn hiu
singlet cng hng ti H 3,49 ppm v mt ni i b th 3 ln vi tn hiu cng
hng ca mt proton ti H 5,25 ppm. T ph ny cn quan st c tn hiu ca
4 proton anomeric ca 4 gc ng, H 4,33 (1H, d, J = 7,5 Hz); H 4,35 (1H, d, J
=7,5 Hz); H 4,38 (1H, d, J = 8,0 Hz); H 4,49 (1H, d, J = 7,5 Hz). Trong vng t
3,03 n 3,8 ppm l cc tn hiu multi ca cc proton ca cc nhm CH; CH2 ni
vi oxy thuc cc gc ng.
OH

21
22

O
HO
19

10

4'

OH

6
1'

NaO3SO

28

29

2'

6'' CH3
O O
4''
6'''

6''''

1''''

OH

2''

O O

O
4''''

1''

OH

CH2OH

CH2OH

OH

4'''

1'''

OCH3
2''''

OH

OH

17

2'''

OH

295

26
25

23

OH
27

15
30

O O

20

13

3
5'

O
18

12

11

24

Hnh IV.1.4.5.b. Cu trc ho hc ca HS5

Hnh IV.1.4.5.c. Ph 13C-NMR ca hp cht HS5

Ph 13C-NMR v cc ph DEPT ca HS5 cho thy s c mt ca 54
cacbon trong phn t, bao gm 9 nhm metyl; 11 nhm metylen; 26 nhm metin
v 8 nguyn t C khng lin kt vi hydro. Pha trng yu c mt nhm
cacbonyl C 173,7 ppm v mt ni i kiu >C=CH- [C 152,63(s)/ 114,5(d)]. S
c mt ca 4 gc ng cng c khng nh vi tn hiu ca 4 cacbon
anomeric (C 103,86; 103,86; 103,86; 102,96 ppm). Ngoi ra cn c 24 cacbon
khc cng lin kt trc tip vi oxy, bao gm mt nhm metoxy (C 59,89 ppm);
3 nhm oxymetylen (C 60,89; 60,77; 63,02 ppm); C bc 4 (C 86,6; 88,2 ppm) v
18 nhm oxymetin (C 68,41; 69,30; 70,15; 72,15; 73,52; 74,27; 74,27; 74,34;
74,85; 76,07; 76,77; 81,44; 85,94; 86,18; 87,92; 87,57; 70,15; 74,8). Nhng
thng tin trn cho thy cu trc ca HS5 kh ging vi HS-3 v HS-4.

296

Hnh IV.1.4.5.d: Cc ph DEPT ca hp cht HS5

Hnh IV.1.4.5.e. Ph HSQC ca hp cht HS5

ghp chnh xc chuyn dch ho hc ca HS5, chng ti tin
hnh o ph cng hng t hai chiu HSQC. So snh d liu ph ca HS5 vi
Holothurin A2 chng ti nhn thy hu ht cc v tr u trng nhau , c th cc v
tr t C-1 n C-21; C-28 n C-30 ca khung Holasthan v chui 4 gc ng
Xyl-Qui-Glc-OmeGlc ca 2 phn t ny u trng khp. S khc bit gia 2 phn
t ny nm 6 v tr t C-22 n C-27.
Bng IV.1.4.5.a: Kt qu ph NMR phn khung Holasthane ca HS5
STT

Ca, b
HS3

Ca, b
HS5

1
2
3
4
5
6
7
8
9

35,8
26,1
87,5
38,9
51,9
20,3
27,1
40,0
152,6

35,67
26,2
87,57
38,33
51,93
20,4
27,1
40,0
152,63

Ha, c
HS5

1,36 - 1,75 (2H, m)

1,65 - 1,85 (2H, m)
3,35 (1H, m)
0,86 (1H, m)
1,45 - 1,67 (2H, m)
1,34 - 1,67 (2H, m)
2,99 (1H, m)

297

HMBC

C-1 Xyl

10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30

39,8
114,5
70,1
57,4
45,3
35,6
34,7
88,13
173,6
21,9
86,2
22,4
32,3*
31,5*
77,4*
34,1*
19,7*
17,7*
16,2
27,4
19,2

39,8
114,5
70,15
57,46
45,38
35,7
34,8
88,2
173,7
21,9
86,6
22,53
34,3
28,2
74,8
33,2
17,7
18,99
16,17
27,45
19,30

5,25
4,43 (1H, m)

C-8

1,05 - 1,70 (2H, m)

1,93 - 2,45 (2H, m)

C-16

1,03 (3H, s)

C-5, C-9, C-10

1,42 (3H, s)
1,61 -1,92 (2H, m)
1,22 - 1,40
3,14 (1H, m)
1,55 (1H m)
0,83 (3H, s)
0,84 (3H, s)
0,81 (3H, s)
1,03 (3H, s)
1,20 (3H, s)

C-17, C-20, C-22

C-24, C-25, C-27

C-24, C-25, C-26
C-3, C-4, C-5, C-29
C-3, C-4, C-5, C-28
C-8, C-14, C-15, C-13

Nhnh C-22 n C-27 ca Curculigosaponin K [231], ao trong DMSO, b125 MHz, c500 MHz.

Hnh IV.1.4.5.f. Ph HMBC ca hp cht HS5

298

Bng IV.1.4.5.b: Kt qu ph NMR phn chui ng ca HS5

Ca, b
HS5

C
Xyl 1
2
3
4
5
Qui 1
2
3
4
5
6
Glc 1
2
3
4
5
6
OMeGlc 1
2
3
4
5
6
OCH3
a

103,86
81,44
74,85
74,27
63,02
103,86
74,34
74,27
86,18
70,15
17,34
102,96
72,15
87,92
68,41
76,07
60,77
103,86
73,52
85,94
69,30
76,77
60,89
59,89

Ha, c
HS5
4,33 (1H, d, J = 7,5 Hz)
3,35 (1H, m)
3,07 (1H, m)
3,95 (1H, m)
3,21 - 3,94 (2H, m)
4,49 (1H, d, J = 7,5 Hz)
3,52 (1H, m)
3,30 (1H, m)
2,99 (1H, m)
3,33 (1H, m)
1,25 (3H, s)
4,38 (1H, J = 8,0 Hz)
3,21 (1H, m)
3,03 (1H, m)
3,21 (1H, m)
3,31 (1H, m)
3,43 - 3,70 (2H, m)
4,35 (1H, d, J = 7,5 Hz)
3,16 (1H, m)
2,99 (1H, m)
3,14 (1H, m)
3,21 (1H, m)
3,39 - 3,70 (2H, m)
3,49 (3H, s)

HMBC
C-3
C1 Xyl

C-1 Xyl, C-4 Xyl

C-2 Xyl
C-3 Qui
C-2 Qui
C-3, C-5, C-6 Qui, C-1 Glc
C-4 Qui, C-5 Qui
C-4 Qui

C-3 Glc
C-2 OMeGlc, OCH3
C-6, C-5 OMeGlc

C-3 OMeGlc

o trong DMSO, 125 MHz, 500 MHz.

Da vo ph cng hng 2 chiu HMBC, cc v tr t C-22 n C-27

c xc nh (Hnh IV.1.4.5.f. v g). C th H-21 khng ch tng tc vi C-17,
C-20 m cn c tng tc vi C-22. Ngoi ra cc v tr t C-24 n C-27 cng
c khng nh bng mt lot cc tng tc ca H-26 v H-27 vi C-24; C-25;
C-26 v C-27. i vi v tr 24 cn quan st c tng tc ca OH-24 (H 4,58
ppm) vi C-24. Thm vo , cc d liu ph 13C-NMR ca phn nhnh t C-22
n C-27 ny cng rt trng khp vi phn nhnh t C-22 n C-27 thuc khung
tritecpen ca hp cht curculigosaponin K [231].
Trt t lin kt trn chui ng cng nh cc v tr trn khung Holasthan
cng c khng nh bng mt lot cc tng tc xa c ch ra trong bng
IV.1.4.5.a v hnh IV.1.4.5.g.

299

OH

21
22

O
HO
19

10

4'

6''

23

OH
27

9
15

OH

13

26
25

30

O O

6
1'

NaO3SO

20
17

3
5'

O
18

12

11

24

28

29

2'

CH3
O O

4''
6'''

6''''

1''''

OH

2''

O O

O
4''''

1''

OH

CH2OH

CH2OH

OH

4'''

1'''

OCH3
2''''

OH

OH

2'''

OH

Hnh IV.1.4.5.g. Mt s tng tc chnh trong phn t HS5

Hnh IV.1.4.5.h. Ph khi lng ESI-MS (positive) ca hp cht HS5

Vi nhng phn tch nu trn, hp cht HS5 c xc nh l mui natri
ca
3,12,17,24-tetrahydroxyholost-9(11)-ene
3-O-[(3-O-methyl)--Dglucopyralnosyl-(13)--D-glucopyranosyl-(14)--D-quinovopyranosyl(12)-(4-O-sunfo)--D-xylopyranoside]. y l mt hp cht mi, ln u tin
c phn lp t thin nhin, v c chng ti t tn l holothurin A4.

300

IV.1.4.6. Hp cht HS6: holothurinogenin B

Hp cht HS6 nhn c di dng cht bt v nh hnh. Bng cc d
kin ph khi lng (ESI-MS: positive ti m/z 762,7 [M+H]+, 785.3 [M+Na]+ v
negative ti m/z 761.3 [M-H]-) v ph cng hng t ht nhn.

Hnh IV.1.4.6.a. Ph 1H-NMR ca hp cht HS6

23

24

21

25
20

18
19

10

13
9

O
OH

15
30

5'
4'

17

12

11

26
22

O O

OH

6
1'

OH

28

29

2'

6'' CH3

O
4''

OH

1''

OH
2''

OH

Hnh IV.1.4.6.b. Cu trc ho hc ca HS6

301

27

Hnh IV.1.4.6.c. Ph 13C-NMR ca hp cht HS6

Ph 1H-NMR ca HS6 cho thy s c mt ca 8 nhm metyl trong phn
t, trong 7 nhm metyl bc 3 vi cc tn hiu singlet (H 0,98; 1,11; 1,12;
1,22; 1,27; 1,31; 1,37) v mt nhm metyl bc 2 cho tn hiu doublet (H 1,28
ppm). vng trng yu c tn hiu cng hng ca 2 ni i b th 3 ln (H
5,55 v 5,30 ppm) v cc tn hiu multi ca cc nhm CH thuc cc gc ng
nm trong vng t 3,00 n 3,53 ppm. Ngoi ra cn quan st c 2 tn hiu
doublet ca 2 proton anome ca 2 gc ng [H 4,24 (1H, d, J = 7,0 Hz)]; H
4,55 (1H, d, J = 7,5 Hz).

Hnh IV.1.4.6.d. Cc ph DEPT ca hp cht HS6

302

Ph 13C-NMR v cc ph DEPT ca HS6 cho thy s c mt ca 41

cacbon trong phn t bao gm 8 nhm CH3; 9 nhm CH2; 14 nhm CH v 10
nguyn t C khng ni vi Hydro. Pha trng yu c tn hiu cng hng ca 1
nhm cacbonyl C 178,3 v 2 ni i kiu >C=CH- [C 142,7(s)/121,0(d)] v [C
148,8(s)/113,2(d)]. Trong vng t 66,4 n 106,0 ppm c 15 nguyn t C c lin
kt vi oxy, trong c 2 cacbon anomeric ca 2 gc ng vi tn hiu c
trng C 105,6 v 106,0 ppm; 3 nguyn t C bc 4; 9 nhm CH; v 1 nhm
oxymetylen. Nhng thng tin trn cho thy hp cht HS6 kh ging vi
Holothurin B, vi cu trc ca 1 khung tritecpen lin kt vi chui 2 phn t
ng.

Hnh IV.1.4.6.e. Ph HSQC ca hp ch HS6

Cc gi tr ph NMR ca cac cacbon c gn vi cc prkoton tng ng
thng qua ph cng hng t ht nhn hai chiu tng tc trc tip HSQC. So
snh d liu ph ca HS6 vi cc hp cht tritecpen glycoside tch c t cc
loi hi sm chng ti nhn thy phn khung holasthan ca HS6 hon ton ph
hp vi hp cht 22,25- oxidoholothurinogenin, mt hp cht tch c t loi
Holothuria leucospilota [232]. Nu so snh vi holothurin B th khung holasthan
ca HS6 c thm mt lin kt v tr C-7, C-8, tuy nhin v tr C-12 khng
cn nhm OH. Hai gc ng trong phn t ny cng c xc nh l Xylo v
Quino, trong gc ng Xylo khng c nhm NaSO3 (so vi trng hp c
nhm NaSO3 th chuyn dch ho hc ca v tr C-4Xylo gim t 74,3 ppm
xung cn 71,1 ppm). D liu ph NMR phn chui ng ca HS6 cng c
i chiu vi hp cht Bivittoside A mt hp cht c phn lp t loi
Bohadschia bivittata [233].

303

Bng IV.1.4.6.a: Kt qu ph NMR phn khung Holasthane ca HS6

1
2
3
4
5
6

Ca, b
[232]
35,9
28,5
88,1
39,4
50,1
25,4

Ca, b
HS6
36,4
28,9
90,6
40,8
51,3
23,9

7
8
9
10
11
12

119,9
142,1
147,9
38,1
112,7
28,4

121,0
142,7
148,8
40,3
113,2
29,7

13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30

57,8
48,7
34,2
28,8
85,8
175,6
18,1
85,5
23,5
81,0
28,9
36,6
81,3
28,7
27,8
16,7
27,8
23,1

58,9
49,6
34,8
27,7
86,9
178,3
23,1
87,1
18,2
81,8
28,5
39,3
82,9
28,9
27,7
17,2
28,6
25,4

Ha, c
HS6
2,8 (2H, m)
2,06 (2H, m)
3,15 (1H, m)

HMBC
C-5
C-1 Xyl; C-28

1,15 (1H, m)
2,13 (2H, dd, 4,5; 6,5 C-7; C-8
Hz)
5,55 (2H, m)
C-6

5,30 (1H, t)
2,25 - 2,85 (2H, m)

C-8; C-10; C-12; C-13

C-9; C-11; C-13; C-14;
C-18

1,36 - 1,86 (2H, m)

1,80 (2H, m)

C-14; C-30; C-13; C-17

1,12 (3H, s)

C-5; C-9; C-10

1,37 (3H, s)
4,21 (1H, t, 7,0 Hz)
2,05 (2H, m)
1,78 - 1,82 (2H, m)

C-20; C-22
C-21; C-20
C-24; C-25
C-23

1,31 (3H, s)
1,27 (3H, s)
0,98 (3H, s)
1,11 (3H, s)
1,22 (3H, s)

C-24; C-25; C-27

C-24; C-25; C-26
C-3; C-4; C-5; C-29
C-3; C-4; C-5; C-28
C-8; C-13; C-14

o trong MeOD, b125 MHz, c500 MHz.

khng nh r trt t lin kt cc v tr trong phn t HS6 chng ti

o ph cng hng t 2 chiu HMBC. T ph ny quan st c cc tng tc xa
gia H-3 vi C-1Xylo; H-1Xylo vi C-3; H-1 Quino vi C-2Xylo. iu ny
cho thy gc ng xylo lin kt vi khung Holasthan v tr C-3, cn gc ng
Quino gn vo v tr C-2 ca ng Xylo. Ngoi ra cn rt nhiu cc tng tc
xa khc, tt c c trnh by trong bng IV.1.4.6.a-b.

304

Hnh IV.1.4.6.f. Ph HMBC ca hp cht HS6

23

24

21

O
12

O
20

18
17

13

11

26
22

19

25

27

OH

1
10

9
15

3
4

5'

4'

6''

30

O O

OH

6
1'

28

29

2'

OH
CH3
O O

4''

OH

1''

OH
2''

OH

Hnh IV.1.4.6.g. Mt s tng tc chnh trong phn t HS6

305

Bng IV.1.4.6.b: Kt qu ph NMR phn chui ng ca HS6

C
Xylo 1
2
3
4
5
Quino 1
2
3
4
5
6

Ca, b
[233]
105,7
84,0
78,1
70,8
66,7
106,2
76,7
77,8
77,1
73,4
18,7

Ca, b
HS6
106,0
83,1
77,8
71,1
66,4
105,6
76,9
77,5
77,0
73,7
18,1

Ha, c
HS6
4,42 (1H, d, 7,0 Hz)
3,49 (1H, m )
3,53 (1H, m)
3,54 (1H, m)
3,23 3,87 (2H, m)
4,55 (1H, d, 7,5 Hz)
3,26 (1H, m )
3,34 (1H, m )
3,00 (1H, m )
3,33 (1H, m )
1,28 (3H, d, 1,66)

HMBC
C-3; C-5 Xyl
C-1; C-3 Xyl
C-2; C-4 Xyl
C-3 Xyl
C-1; C-3; C-4
C-2 Xyl; C-5 Qui
C-1; C-3 Qui
C-4; C-1 Qui
C-6; C-3; C-5 Qui
C-5; C-4 Qui

o trong MeOD, b125 MHz, c500 MHz.

Hnh IV.1.4.6.h. Ph khi lng ESI-MS ca hp cht HS6

Vi nhng phn tch nu trn, hp cht HS6 c nhn dng l 3-O-[D-quinovopyranosyl-(12)--D-xylopyranosil]-22,25-epoxyholosta-7,9(11)diene-17-ol. y l mt cht mi, ln u tin c phn lp t thin nhin, v
c chng ti t tn l holothurinogenin B.

306

4.305
3.551
3.037
3.012
2.988
2.575
2.555
2.538
2.521
2.514
2.496
2.487
2.284
2.261
2.186
2.164
2.138
2.054
2.024
1.992
1.970
1.932
1.911
1.894
1.881
1.868
1.855
1.716
1.696
1.689
1.682
1.674
1.657
1.573
1.556
1.524
1.485
1.472
1.459
1.446
1.432
1.302
1.291
1.266
1.254
1.241
1.222
1.203
1.185
1.163
1.100
1.063
1.050
0.955
0.941
0.864
0.856
0.851

5.265
5.251
5.235
5.220
5.200
5.184
5.169
5.154
4.910
4.813

5.725

5.965

6.237

7.190

7.558

8.707

IV.1.4.7. Hp cht HS7: 2,3,12,21,24R,25-hexahydroxylanosta-8-ene

Hp cht HS7 c tch ra t dch chit CHCl3, c dng tinh th hnh que,
mu trng. T kt qu ph khi lng ESI-MS (m/z 490 [M-H2O]+) v ph cng
hng t ht nhn cho php xc nh cng thc phn t ca HS7 l C30H52O6 (M
508).

Current Data Parameters

NAME
jn3
EXPNO
1
PROCNO
1
F2 - Acquisition Parameters
Date_
20060323
Time
16.45
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
zg
TD
32768
SOLVENT
Pyr
NS
8
DS
0
SWH
6009.615 Hz
FIDRES
0.183399 Hz
AQ
2.7264309 sec
RG
64
DW
83.200 usec
DE
6.00 usec
TE
297.6 K
D1
2.00000000 sec
MCREST
0.00000000 sec
MCWRK
0.01500000 sec
======== CHANNEL f1 ========
NUC1
1H
P1
5.00 usec
PL1
-3.00 dB
SFO1
500.1321372 MHz
F2 - Processing parameters
SI
16384
SF
500.1300004 MHz
WDW
no
SSB
0
LB
0.00 Hz
GB
0
PC
1.00

0.5

ppm

9.1589

1.0

3.0327

1.5

3.2145
13.4125

2.0

7.1138

2.5

14.3784

3.0

4.5886

3.5

5.2438

4.0

1.0576
1.1167

4.5

1.9433

5.0

1.0000

5.5

1.0060

6.0

1.0910
1.0078

6.5

2.4675

7.0

0.9668

7.5

0.5425

8.0

0.5793

8.5

Hnh IV.1.4.7.a. Ph 1H-NMR ca HS7

Trn ph 1H-NMR xut hin tn hiu cng hng ca 7 nhm metyl bc 3
ti 0,65, 0,82, 1,02, 1,04, 1,06, 1,15 v 1,17 ppm. Bn cnh , vng trng
yu cn c cc tn hiu ca 4 nhm oximetyl vi 2,93 ppm (1H, dd, J =9,0,
1,5Hz), 3,25ppm (1H, dd, 9,5, 2,0Hz), 3,63 (1H, m), 4,0 (1H, dd, J = 8,5 v
1,7Hz) v 1 nhm oximetylen ti 3,71ppm (2H, dd, J = 12,5 v 2,5Hz).
OH

21

OH

18

12
11

HO

10

HO
H
28

OH

15

25

16

14

26

27

17

24
23

20

13

19
1

22

OH

30

29

Hnh IV.1.4.7.b. Cu trc ha hc ca HS7

307

79.436
74.369
73.835
69.876
61.961
51.978
51.223
51.083
49.514
49.344
49.174
49.004
48.834
48.664
48.494
44.834
44.274
40.288
39.208
39.141
33.249
32.841
29.403
29.113
28.856
28.046
27.573
25.643
25.006
24.147
20.341
19.392
17.406
17.247

84.273

136.330
134.332

Current Data Parameters

NAME
jn2
EXPNO
2
PROCNO
1
F2 - Acquisition Parameters
Date_
20060323
Time
1.28
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
zgdc
TD
65536
SOLVENT
DMSO
NS
6144
DS
0
SWH
35211.270 Hz
FIDRES
0.537281 Hz
AQ
0.9306754 sec
RG
16384
DW
14.200 usec
DE
6.00 usec
TE
298.8 K
D1
2.00000000 sec
d11
0.03000000 sec
MCREST
0.00000000 sec
MCWRK
0.01500000 sec
======== CHANNEL f1 ========
NUC1
13C
P1
10.00 usec
PL1
-5.00 dB
SFO1
125.7716224 MHz
======== CHANNEL f2 ========
CPDPRG2
waltz16
NUC2
1H
PCPD2
80.00 usec
PL2
120.00 dB
PL12
15.00 dB
SFO2
500.1323100 MHz
F2 - Processing parameters
SI
32768
SF
125.7576149 MHz
WDW
EM
SSB
0
LB
3.00 Hz
GB
0
PC
1.00

220

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

Hnh IV.1.4.7.c. Ph 13C-NMR ca HS7

Bng IV.1.4.7. Kt qu ph NMR ca HS7
C

Ca [234]

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21

44,4 (t)
68,9 (t)
83,5 (d)
39,6 (s)
51,0 (d)
18,7 (t)
28,3 (t)
134,7 (s)
133,9 (s)
38,4 (s)
26,8 (t)
73,1 (d)
50,6 (s)
50,2 (s)
32,3 (t)
29,7 (t)
38,6 (d)
17,1 (q)
20,3 (q)
44,0 (d)
61,2 (t)

44,8 (t)
69,9 (d)
84,3 (d)
40,3 (s)
51,9 (d)
19,4 (t)
28,8 (t)
134,3 (s)
133,4 (s)
39,1 (s)
27,6 (t)
74,4 (d)
51,2 (s)
51,0 (s)
32,8 (t)
29,4 (t)
39,2 (d)
17,2 (q)
20,3 (q)
44,2 (d)
61,9 (t)

22
23
24
25

28,1 (t)
33,1 (t)
78,9 (d)
72,6 (s)

28,0 (t)
33,2 (t)
79,4 (d)
73,8 (s)

Cab

Hac

HMBC

1,13 (m)/2,04 (m)

3,63 (1H, m)
2,93 (1H, dd, 1,5, 9,0)
1,12 *
1,54 (m)/2,10*
1,45 (m)/2,21*
2,09*
4,00 (1H, dd, 1,7, 8,5)
1,18 (m)/1,76 (m)
1,35 (m)/1,43 (m)
2,41 (1H, m)
0,65 (3H, s)
1,04 (3H, s)
1,38 (1H, m)
3,71 (dd, 2,5, 12,5)
3,79 (dd, 2,5, 12,5)
2,10*
2,00 (m)/2,75 (m)
3,25 (1H, dd, 2,0, 9,5)
-

308

C-4, 5, 28, 29

C-9, 13, 14

C-12, 13, 14, 17

C-1, 5, 9, 10
C-17, 20, 22
C-17, 20, 22

C-23, 25

26
27
28
29
30

26,0 (q)
25,7 (q)
17,4 (q)
29,0 (q)
24,1 (q)

25,6 (q)
25,0 (q)
17,4 (q)
29,1 (q)
24,1 (q)

1,17 (3H, s)
1,15 (3H, s)
0,82 (3H, s)
1,02 (3H, s)
1,06 (3H, s)

C-24, 25, 27
C-24, 25, 26
C-4, 29, 3, 5
C-4, 28, 3, 5
C-8

o trong Py-d5, b125MHz, c500MHz, *tn hiu b che lp.

Trn ph 13C-NMR xut hin tn hiu ca 30 cacbon trong phn t gi

cho mt hp cht tritecpen. Thng qua ph DEPT cho php xc nh s c mt
ca 7 nhm CH3, 9 nhm CH2, 7 nhm CH v 7 cacbon bc 4. Trong c 4
nhm oximetyl, 1 nhm oximetylen, v 1 nguyn t cacbon bc 4 lin kt trc
tip vi oxi c khng nh tng ng bi cc tn hiu cng hng ti 69,9
(CH, C-2), 84,3 (CH, C-3), 74,4 (CH, C-12), 79,4 (CH, C-24), 61,9 (CH2, C-21)
v 73,8 (C, C-25). Cc tn hiu cng hng ti 134,3 (C, C-8) v 133,4 (C, C-9)
khng nh s tn ti ca mt lin kt i b th hon ton.
Current Data Parameters
NAME
jn2hmqc
EXPNO
1
PROCNO
1

ppm

20

30

40

50

60

70

80

90

100

110

120

F2 - Acquisition Parameters
Date_
20060322
Time
18.32
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
invbph
TD
1024
SOLVENT
MeOD
NS
4
DS
16
SWH
3004.808 Hz
FIDRES
2.934382 Hz
AQ
0.1706100 sec
RG
228.1
DW
166.400 usec
DE
6.00 usec
TE
297.9 K
CNST2
145.0000000
d0
0.00000300 sec
D1
1.79999995 sec
d2
0.00344828 sec
D7
0.28735110 sec
IN0
0.00001807 sec
MCREST
0.00000000 sec
MCWRK
1.79999995 sec
======== CHANNEL f1 ========
NUC1
1H
P1
10.20 usec
p2
20.40 usec
PL1
-3.00 dB
SFO1
500.1312503 MHz
======== CHANNEL f2 ========
CPDPRG2
garp
NUC2
13C
P3
18.50 usec
p4
37.00 usec
PCPD2
65.00 usec
PL2
-5.00 dB
PL12
5.00 dB
SFO2
125.7716224 MHz
F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7716 MHz
FIDRES
108.056709 Hz
SW
219.942 ppm
FnMODE
TPPI
F2 - Processing parameters
SI
1024
SF
500.1300161 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
TPPI
SF
125.7576149 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

130

4.0

3.5

3.0

2.5

2.0

1.5

1.0

0.5

140
ppm

Hnh IV.1.4.7.d. Ph HSQC ca HS7

S liu ph NMR ca cc cacbon c gn vi cc proton tng ng thng
qua ph cng hng t ht nhn hai chiu tng tc trc tip HSQC. Kt qu
c tng hp trong bng IV.1.4.7.

309

Current Data Parameters

NAME
jn2hmbc
EXPNO
1
PROCNO
1

ppm
10

20

30

40

50

60

70

80

90

100

110

120

130

140

4.0

3.5

3.0

2.5

2.0

1.5

1.0

0.5

ppm

F2 - Acquisition Parameters
Date_
20060322
Time
19.12
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
hmbcgplpndqf
TD
1024
SOLVENT
MeOD
NS
8
DS
16
SWH
3004.808 Hz
FIDRES
2.934382 Hz
AQ
0.1706100 sec
RG
10321.3
DW
166.400 usec
DE
6.00 usec
TE
299.8 K
CNST2
145.0000000
CNST13
7.0000000
d0
0.00000300 sec
D1
1.79999995 sec
d2
0.00344828 sec
d6
0.07142857 sec
D16
0.00020000 sec
IN0
0.00001656 sec
MCREST
0.00000000 sec
MCWRK
1.79999995 sec
======== CHANNEL f1 ========
NUC1
1H
P1
10.20 usec
p2
20.40 usec
PL1
-3.00 dB
SFO1
500.1312503 MHz
======== CHANNEL f2 ========
NUC2
13C
P3
18.50 usec
PL2
-5.00 dB
SFO2
125.7728799 MHz
====== GRADIENT CHANNEL
GPNAM1
sine.100
GPNAM2
sine.100
GPNAM3
sine.100
GPX1
0.00
GPX2
0.00
GPX3
0.00
GPY1
0.00
GPY2
0.00
GPY3
0.00
GPZ1
50.00
GPZ2
30.00
GPZ3
40.10
P16
1000.00

=====

%
%
%
%
%
%
%
%
%
usec

F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7729 MHz
FIDRES
117.924530 Hz
SW
240.025 ppm
FnMODE
QF
F2 - Processing parameters
SI
1024
SF
500.1300161 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
QF
SF
125.7576149 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0

Hnh IV.1.4.7.e. Ph HMBC ca HS7

ppm
0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0
5.0

4.5

4.0

3.5

3.0

2.5

2.0

1.5

1.0

Hnh IV.1.4.7.f. Ph H-H COSY ca HS7

310

0.5

ppm

Current Data Parameters

NAME
jn2cosy
EXPNO
1
PROCNO
1
F2 - Acquisition Parameters
Date_
20060322
Time
18.02
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
cosygpqf
TD
1024
SOLVENT
MeOD
NS
4
DS
16
SWH
3004.808 Hz
FIDRES
2.934382 Hz
AQ
0.1706100 sec
RG
128
DW
166.400 usec
DE
6.00 usec
TE
298.2 K
d0
0.00000300 sec
D1
1.50000000 sec
d13
0.00000400 sec
D16
0.00020000 sec
IN0
0.00033325 sec
MCREST
0.00000000 sec
MCWRK
1.50000000 sec
======== CHANNEL f1 ========
NUC1
1H
P0
5.10 usec
P1
10.20 usec
PL1
-3.00 dB
SFO1
500.1312503 MHz
====== GRADIENT CHANNEL =====
GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
10.00 %
GPZ2
10.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
256
SFO1
500.1313 MHz
FIDRES
11.721681 Hz
SW
6.000 ppm
FnMODE
QF
F2 - Processing parameters
SI
1024
SF
500.1300161 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0
PC
4.00
F1 - Processing parameters
SI
1024
MC2
QF
SF
500.1300161 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0

OH

OH

OH
OH

HO

HO
H

Hnh IV.1.4.7.g. Cc tng tc HMBC chnh ca HS7

Phn tch chi tit cc tng tc trn ph H-H COSY cho php ni cc
mnh cu trc t C-1 n C-4, C-6 sang C-7, C-11 sang C-12, C-15 n C-18 v
t C-18 n C-24. Kt qu ny, cng vi cc tng tc xa H-C trn ph HMBC
(hnh IV.1.4.7.g.) cho php gn kt cc mnh cu trc li vi nhau v xy dng
nn cu trc ton b phn t ca HS7 nh c ch ra hnh IV.1.4.7.b. Ngoi
ra, s ph hp hon ton v s liu ph 13C-NMR ca HS7 so vi cc s liu
c cng b (bng IV.1.4.7) [234] cho php khng nh cu trc ha hc ca n
l 2,3,12,21,24R,25-hexahydroxylanosta-8-ene, y l ln u tin hp cht
ny c tch t loi Holothuria scabra.

4.305
3.551
3.037
3.012
2.988
2.575
2.555
2.538
2.521
2.514
2.496
2.487
2.284
2.261
2.186
2.164
2.138
2.054
2.024
1.992
1.970
1.932
1.911
1.894
1.881
1.868
1.855
1.716
1.696
1.689
1.682
1.674
1.657
1.573
1.556
1.524
1.485
1.472
1.459
1.446
1.432
1.302
1.291
1.266
1.254
1.241
1.222
1.203
1.185
1.163
1.100
1.063
1.050
0.955
0.941
0.864
0.856
0.851

5.265
5.251
5.235
5.220
5.200
5.184
5.169
5.154
4.910
4.813

5.725

5.965

6.237

7.190

7.558

8.707

IV.1.4.8. Hp cht HS8: 24R- ergosta-7,22-dien-3,5,6-triol

T phn on CHCl3 chng ti phn lp c hp cht HS8 di dng
tinh th hnh que, mu trng. Cng thc phn t ca n c xc nh l C28H46O3
bng cc kt qu ph khi lng (m/z 412 [M-H2O]+) v ph cng hng t ht
nhn.
Current Data Parameters
NAME
jn3
EXPNO
1
PROCNO
1
F2 - Acquisition Parameters
Date_
20060323
Time
16.45
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
zg
TD
32768
SOLVENT
Pyr
NS
8
DS
0
SWH
6009.615 Hz
FIDRES
0.183399 Hz
AQ
2.7264309 sec
RG
64
DW
83.200 usec
DE
6.00 usec
TE
297.6 K
D1
2.00000000 sec
MCREST
0.00000000 sec
MCWRK
0.01500000 sec
======== CHANNEL f1 ========
NUC1
1H
P1
5.00 usec
PL1
-3.00 dB
SFO1
500.1321372 MHz
F2 - Processing parameters
SI
16384
SF
500.1300004 MHz
WDW
no
SSB
0
LB
0.00 Hz
GB
0
PC
1.00

Hnh IV.1.4.8.a. Ph 1H-NMR ca HS8

311

9.1589

0.5
3.0327

1.0
3.2145

1.5

13.4125

2.0

7.1138

2.5

4.5886

3.0

14.3784

3.5

5.2438

4.0

1.0576
1.1167

4.5

1.9433

5.0

1.0000

5.5

1.0060

6.0

1.0910
1.0078

6.5

2.4675

7.0

0.9668

7.5

0.5425

8.0

0.5793

8.5

ppm

28
21

22

18

24
23

20

12

13
14

10

15

HO

16

2
3

27

17

11
19

26
25

7
6

OH

OH

Hnh IV.1.4.8.b. Cu trc ho hc ca HS8

56.158
55.252
43.758
43.080
41.948
40.838
39.914
38.067
33.825
33.345
32.616
32.137
29.998
29.618
28.459
23.474
22.946
22.408
21.402
20.140
19.836
18.781
17.830
14.279
12.511
1.422

67.553

76.108
75.999
74.256

150.229
150.109
149.893
149.678
141.554
136.189
135.795
135.702
135.504
135.307
132.115
123.789
123.698
123.500
123.302
120.465

Trn ph 1H-NMR c trng cho mt hp cht sterol trong xut hin 2

tn hiu singlet ca 2 nhm metyl bc 3 vi 0,65, H 1,52ppm v cc tn hiu
doublet ca 4 nhm metyl bc 2 ti 0,86 (6H, d, J = 6,5 Hz), 0,94 (3H, d,
6,5Hz), H 1,07 (3H, d, 6,5Hz). Du hiu trn c trng cho mt hp cht c cu
trc b khung dng ergosterol. vng trng yu c tn hiu cng hng ca 2
nhm oximetyl ti H 4,28; H 4,70ppm v tn hiu ca 2 proton olefinic ca 1
n i b th hai v tr, cu hnh trans vi 5,18 (1H, m) v 5,22 (1H, dd, J =
2,5, 15,0 Hz). Ngoi ra, trong phn t HS8 cn 1 ni i b th 3 v tr vi tn
hiu cng hng 5,72 ppm (1H, m).

Current Data Parameters

NAME
jn3
EXPNO
2
PROCNO
1
F2 - Acquisition Parameters
Date_
20060323
Time
21.28
INSTRUM
spect
PROBHD 5 mm TBI 1H-13
PULPROG
zgdc
TD
65536
SOLVENT
DMSO
NS
6144
DS
0
SWH
35211.270 Hz
FIDRES
0.537281 Hz
AQ
0.9306754 sec
RG
16384
DW
14.200 usec
DE
6.00 usec
TE
298.6 K
D1
2.00000000 sec
d11
0.03000000 sec
MCREST
0.00000000 sec
MCWRK
0.01500000 sec
======== CHANNEL f1 ========
NUC1
13C
P1
10.00 usec
PL1
-5.00 dB
SFO1
125.7716224 MHz
======== CHANNEL f2 ========
CPDPRG2
waltz16
NUC2
1H
PCPD2
80.00 usec
PL2
120.00 dB
PL12
15.00 dB
SFO2
500.1323100 MHz
F2 - Processing parameters
SI
32768
SF
125.7577579 MHz
WDW
EM
SSB
0
LB
3.00 Hz
GB
0
PC
1.00

220

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

Hnh IV.1.4.8.c. Ph 13C-NMR ca HS8

312

10

ppm

Ph 13C-NMR v DEPT cho thy s c mt ca 28 cacbon trong phn t,

bao gm 6 nhm metyl, 7 nhm metylen, 11 nhm metin v 4 cacbon bc 4. S
c mt ca 2 lin kt cng c ghi nhn. Cc tn hiu C 120,4 (CH), 141,5 (C)
l ca lin kt >C=CH-, cn lin kt -CH=CH- c C 132,0 v 136,2 ppm. Bn
cnh cc tn hiu ca 2 nhm oximetin C 67,5 v 74,2 cn c 1 nguyn t cacbon
bc 4 lin kt trc tip vi oxi ti 76,1ppm.
Current Data Parameters
NAME
jn3hmqc
EXPNO
1
PROCNO
1

ppm
10

20

30

40

50

60

70

80

90

100

110

120

F2 - Acquisition Parameters
Date_
20060323
Time
17.31
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
invbph
TD
1024
SOLVENT
MeOD
NS
4
DS
16
SWH
4595.588 Hz
FIDRES
4.487879 Hz
AQ
0.1115700 sec
RG
181
DW
108.800 usec
DE
6.00 usec
TE
297.6 K
CNST2
145.0000000
d0
0.00000300 sec
D1
1.79999995 sec
d2
0.00344828 sec
D7
0.38835120 sec
IN0
0.00001807 sec
MCREST
0.00000000 sec
MCWRK
1.79999995 sec
======== CHANNEL f1 ========
NUC1
1H
P1
10.20 usec
p2
20.40 usec
PL1
-3.00 dB
SFO1
500.1317241 MHz
======== CHANNEL f2 ========
CPDPRG2
garp
NUC2
13C
P3
18.50 usec
p4
37.00 usec
PCPD2
65.00 usec
PL2
-5.00 dB
PL12
5.00 dB
SFO2
125.7716224 MHz
F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7716 MHz
FIDRES
108.056709 Hz
SW
219.942 ppm
FnMODE
TPPI
F2 - Processing parameters
SI
1024
SF
500.1300000 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
TPPI
SF
125.7577579 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

130

140
6.0

5.5

5.0

4.5

4.0

3.5

3.0

2.5

2.0

1.5

1.0

0.5

ppm

Hnh IV.1.4.8.d. Ph HSQC ca HS8

Tp hp cc thng tin trn, cho php nhn nh hp cht HS8 l mt sterol
c dng khung ergosterol c cha mt lin kt i ni vng b th 3 v tr, mt
lin kt i ngoi vng b th 2 v tr c cu hnh trans v c cha 3 nhm OH
trong phn t.
Bng IV.1.4.8. Kt qu ph NMR ca HS8
C

Ca [235]

Cab

Hac (J, Hz)

H-H COSY

1
2
3
4
5
6
7
8

33,9 (t)
32,6 (t)
67,6 (d)
41,9 (t)
76,1 (s)
74,2 (d)
120,4 (d)
141,6 (s)

33,9 (t)
32,6 (t)
67,5 (d)
41,9 (t)
76,1 (s)
74,2 (d)
120,4 (d)
141,5 (s)

1,71 (m)/2,16 (m)

1,97 (m)/2,26 (m)
4,70 (1H, m)
2,50 (m)/3,00 (m)
4,28 (1H, br s)
5,72 (1H, br s)
-

H-4
H-3
H-7
H-6
-

313

HMBC
H-C

C-2, 3, 5
C-10, 7, 8
C-5, 9, 14
-

9
10
11
12
13
14

43,8 (d)
38,1 (s)
22,4 (t)
39,9 (t)
43,8 (s)
55,3 (d)

43,8 (d)
38,0 (s)
22,4 (t)
39,9 (t)
43,8 (s)
55,2 (d)

2,54 (1H, m)
1,68 (m)
1,30 (m)/2,06 (m)
1,92 (1H, m)*

H-11
H-9

15
16
17
18

23,5 (t)
28,5 (t)
56,2 (d)
12,5 (q)

23,4 (t)
28,4 (t)
56,1 (d)
12,5 (q)

1,54 *
1,67*
1,23*
0,65(3H, s)

19
20
21
22
23

18,8 (q)
40,8 (d)
21,5 (q)
136,2 (d)
132,1 (d)

18,7 (q)
1,52 (3H, s)
40,8 (d)
2,04 (1H, m)
H-17, 21, 22
21,4 (q)
1,07 (3H, d, 6,5)
H-20
136,2 (d) 5,18 (1H, dd, 3,0, 15,0)
H-20, 23
132,0 (d) 5,22 (1H, dd, 2,5, 15,0)
H-22, 24

24
25
26
27
28

43,1 (d)
33,4 (d)
20,2 (q)
19,8 (q)
17,8 (q)

43,1 (d)
33,4 (d)
20,1 (q)
19,8 (q)
17,8 (q)

C-11, 17
C-7, 8, 9, 15,
17

H-20

1,86 (1H, m)
1,46 (1H, m)
0,86 (3H, d, 6,5)
0,86 (3H, d, 6,5)
0,94 (3H, d, 6,5)

C-16, 18
C-12, 13, 14,
17
C-1, 9, 10
C-17, 20, 22
C-21, 23
C-20, 22, 24,
25

H-23, 25, 28
H-24, 26
H-25
H-25
H-24

C-25, 24
C-25, 24
C-23, 24

Measured in Py-d5, b125MHz, c500MHz

ppm

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5
6.5

6.0

5.5

5.0

4.5

4.0

3.5

3.0

2.5

2.0

1.5

1.0

0.5

Hnh IV.1.4.8.e. Ph H-H COSY ca HS8

314

ppm

Current Data Parameters

NAME
jn3cosy
EXPNO
1
PROCNO
1
F2 - Acquisition Parameters
Date_
20060323
Time
17.02
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
cosygpqf
TD
1024
SOLVENT
MeOD
NS
4
DS
16
SWH
4595.588 Hz
FIDRES
4.487879 Hz
AQ
0.1115700 sec
RG
128
DW
108.800 usec
DE
6.00 usec
TE
297.7 K
d0
0.00000300 sec
D1
1.50000000 sec
d13
0.00000400 sec
D16
0.00020000 sec
IN0
0.00021760 sec
MCREST
0.00000000 sec
MCWRK
1.50000000 sec
======== CHANNEL f1 ========
NUC1
1H
P0
5.10 usec
P1
10.20 usec
PL1
-3.00 dB
SFO1
500.1317241 MHz
====== GRADIENT CHANNEL =====
GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
10.00 %
GPZ2
10.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
256
SFO1
500.1313 MHz
FIDRES
17.951517 Hz
SW
9.189 ppm
FnMODE
QF
F2 - Processing parameters
SI
1024
SF
500.1300004 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0
PC
4.00
F1 - Processing parameters
SI
1024
MC2
QF
SF
500.1300004 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0

Current Data Parameters

NAME
jn3hmbc
EXPNO
1
PROCNO
1

ppm
10

20

30

40

50

60

70

80

90

100

110

120

130

140

150
6.5

6.0

5.5

5.0

4.5

4.0

3.5

3.0

2.5

2.0

1.5

1.0

0.5

ppm

F2 - Acquisition Parameters
Date_
20060323
Time
18.30
INSTRUM
spect
PROBHD
5 mm TBI 1H-13
PULPROG
hmbcgplpndqf
TD
1024
SOLVENT
MeOD
NS
8
DS
16
SWH
4595.588 Hz
FIDRES
4.487879 Hz
AQ
0.1115700 sec
RG
16384
DW
108.800 usec
DE
6.00 usec
TE
297.6 K
CNST2
145.0000000
CNST13
7.0000000
d0
0.00000300 sec
D1
1.79999995 sec
d2
0.00344828 sec
d6
0.07142857 sec
D16
0.00020000 sec
IN0
0.00001656 sec
MCREST
0.00000000 sec
MCWRK
1.79999995 sec
======== CHANNEL f1 ========
NUC1
1H
P1
10.20 usec
p2
20.40 usec
PL1
-3.00 dB
SFO1
500.1317241 MHz
======== CHANNEL f2 ========
NUC2
13C
P3
18.50 usec
PL2
-5.00 dB
SFO2
125.7728799 MHz
====== GRADIENT CHANNEL
GPNAM1
sine.100
GPNAM2
sine.100
GPNAM3
sine.100
GPX1
0.00
GPX2
0.00
GPX3
0.00
GPY1
0.00
GPY2
0.00
GPY3
0.00
GPZ1
50.00
GPZ2
30.00
GPZ3
40.10
P16
1000.00

=====

%
%
%
%
%
%
%
%
%
usec

F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7729 MHz
FIDRES
117.924530 Hz
SW
240.025 ppm
FnMODE
QF
F2 - Processing parameters
SI
1024
SF
500.1300004 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
QF
SF
125.7577579 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0

Hnh IV.1.4.8.f. Ph HMBC ca HS8

HO
OH
OH

Hnh IV.1.4.8.g. Cc tng tc HMBC chnh ca HS8

T cc kt qu trn kt hp vi phn tch chi tit cc tng tc trn ph
HSQC, H-H COSY v cc tng tc xa C-H trn ph HMBC (Bng IV.1.4.8 v
hnh IV.1.4.8.f-g) cho php ghp ni cc phn cu trc v xy dng nn cu trc
tng th ca ton b phn t HS8 nh c ch ra trn hnh IV.1.4.8.b. Ngoi ra,
s ph hp hon ton v gi tr ph 13C-NMR ti cc v tr tng ng ca HS8 so
vi cc s liu c cng b (Bng IV.1.4.8) [235] cho php khng nh cu
trc ha hc ca hp cht ny l 24R-ergosta-7,22-dien-3,5,6-triol. Hp cht
ny c phn lp t mt s loi san h mm [235] v t nm Agaricus blazei
[236], v Lentinus edodes [237]. Tuy nhin, y l ln u tin n c phn lp
t loi hi sm Holothuria scabra.

315

IV.1.4.9. Hp cht HS-9: -Adenosine

Hp cht HS9 nhn c di dng tinh th hnh kim mu trng. Cng
thc phn t ca n c xc nh l C10H13N5O4 da trn kt qu ph khi lng
ESI-MS (ti m/z 267,8 [M+H]+ v 289,8 [M+Na]+) v ph cng hng t ht nhn
(1H, 13C-NMR v DEPT).
NH2
N
8

6
1

2
3

HO
5'

O
H

4'

3'

OH

1'

H
2'

OH

Hnh IV.1.4.9.a. Cu trc ha hc ca HS9

Hnh IV.1.4.9.b. Ph 1H-NMR ca HS9

Trn ph 1H-NMR ca HS9 xut hin cc tn hiu c trng ca mt n v
ng -D-Ribofuranosyl ti 5,88 (d, J = 6,5 Hz, H-1), 4,61 (dd, J = 4,0 v
10,5 Hz, H-2), 4,15 (br s, H-3), 3,97 (dd, J = 3,5 v 6,5 Hz, H-4), 3,57 (m, Ha5) v 3,68 (m, Hb-5).

316

Bng IV.1.4.9. Kt qu ph NMR ca HS9

Ha, c
(multiplicity, J in Hz)
2
152,5 d
152,41 d
8,14 (s)
4
149,1 s
149,08 s
5
119,4 s
119,37 s
6
156,2 s
156,17 s
8
140,0 d
139,95 d
8,34 (s)
1
88,1 d
87,95 d
5,88 (d, 6,5)
2
73,6 d
73,47 d
4,61 (dd, 4,0, 10,5)
3
70,8 d
70,68 d
4,15 (br s)
4
86,1 d
85,92 d
3,97 (dd, 3,5, 6,5)
61,69 t
3,57 (m)
5
61,8 t
3,68 (m)
NH2
7,32 (s)
OH-2
5,42 *
OH-3
5,17 (d, 3,0)
OH-5
5,43 *
a
o trong DMSO, b125 Mhz, c500MHz
C

C [239]

Ca, b

HMBC (HC)
C-4, 6

C-4, 5
C-4, 8, 2

C-5

Hnh IV.1.4.9.c. Ph 13C-NMR v cc ph DEPT ca HS9

317

Hnh IV.1.4.9.d. Ph HSQC ca HS9

Trn ph 13C-NMR xut hin cc tn hiu ca 10 cacbon vi 3 C, 6 CH v 1
CH2. Trong , cc tn hiu ca 1 n v ng -D-Ribofuranosyl c khng
nh ti 87,95 (d, C-1), 73,47 (d, C-2), 70,68 (d, C-3), 85,92 (d, C-4) v
61,69 (t, C-5) [238].

Hnh IV.1.4.9.e. Ph HSQC ca HS9

318

Hnh IV.1.4.9.f. Ph ESI-MS ca HS9

T cc phn tch nu trn, s liu ph 13C-NMR ca HS9 c mang so
snh vi cc gi tr cng b ca hp cht -Adenosine [239]. S ph hp hon
ton cc gi tr tng ng cho php khng nh cu trc ha hc ca HS9 l 9-D-Ribofuranosyladenine hay -Adenosine. iu ny mt ln na c khng
nh bi cc tng tc trn ph cng hng t ht nhn hai chiu HMBC (Bng
IV.1.4.9 v Hnh IV.1.4.9.e).
IV.1.5. Kt qu nh gi hot tnh sinh hc ca cc cht
IV.1.5.1. Hot tnh gy c t bo ca cc hp cht.
Trc tin, tin hnh nh gi gi tr CS% ca cc hp cht. Gi tr CS%
hay chnh l gi tr biu hin lng t bo sng st sau khi cho tip xc vi cc
mu chit c tnh theo cc cng thc trnh by trong phn quy trnh. Cc
cht c CS% < 50 c coi l c hot tnh v s c tip tc tin hnh xc nh
gi tr IC50.
Kt qu th nghim cho thy: Tt c cc hp cht hothurin u th hin
hot tnh gy c t bo v cc hp cht cn li khng c biu hin hot tnh.
iu ny th hin s ph hp vi cc cng trnh nghin cu c cng b trn
th gii bi lp cht ny c hot tnh gy c t bo c trng. Cc hp cht HS1,
HS2, HS3 v HS4 th hin hot tnh trn c 3 dng t bo th nghim, hp cht
HS5 th hin hot tnh trn hai dng t bo th nghim l Hep-G2 v KB, v hp
cht HS6 ch th hin hot tnh trn dng t bo Hep-G2.

319

Bng IV.1.5.1.a. Kt qu th nghim hot tnh gy c t bo

STT

K hiu
mu

Kt lun

Dng t bo/t l sng st (%)

Hep-G2

KB

FL

DMSO

100,0 0,0

100,0 0,0

100,0 0,0

Ellipticine

2,2 0,0

3,2 0,0

2,05 0,02

Dng tnh

HS-1

21,9 0,39

32,03 0,15

15,17 0,8

Dng tnh c 3 dng

HS-2

23,3 0,09

2,29 0,1

0,0 0,0

Dng tnh c 3 dng

HS-3

12,0 0,03

13,78 0,02

0,0 0,0

Dng tnh c 3 dng

HS-4

42,6 0,14

1,29 0,10

30,0 0,62

Dng tnh c 3 dng

HS-5

22,3 0,31

12,6 0,77

61,2 1,57

Dng tnh 2 dng

HS-6

8,8 0,15

82,1 1,57

83,8 0,62

Dng tnh 1 dng

Bng IV.1.5.1.b. Kt qu IC50 th nghim hot tnh gy c t bo

STT

K hiu
mu

Dng t bo/Gi tr IC50 (g/ml)

Hep-G2

KB

FL

Kt lun

Ellipticine

0,30

0,22

0,15

Dng tnh

HS-1

1,99

2,85

2,14

Dng tnh c 3 dng

HS-2

4,08

1,28

1,54

Dng tnh c 3 dng

HS-3

1,06

2,42

1,00

Dng tnh c 3 dng

HS-4

0,23

0,57

0,39

Dng tnh c 3 dng

HS-5

0,57

1,12

>5

Dng tnh 2 dng

HS-6

2,33

>5

>5

Dng tnh 1 dng

320

Kt qu quan st hnh thi b mt mng t bo

Ngoi ra, 2 hp cht HS-3 v HS-4 c th nghim hot tnh theo phng
php quan st trc tip di knh hin vi. Tc dng gy c t bo ca cc hp
cht HS-3 v HS-4 c quan st v chp nh trn knh hin vi sau cc thi im
24h, 48h, 72h.
Sau 24 gi

Mu i chng m

Nng 5 g/ml

Nng 1 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml
Hnh IV.1.5.1.a: Hnh nh t bo vi mu th l HS-3 cc nng khc nhau
321

Mu i chng m

Nng 5 g/ml

Nng 1 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml

Hnh IV.1.5.1.b: Hnh nh t bo vi mu th l HS-4 cc nng khc nhau

322

- Sau 48 gi

Mu i chng m

Nng 1 g/ml

Nng 5 g/ml

Nng 0,04 g/ml

Nng 0,2 g/ml
Hnh IV.1.5.1.c: Hnh nh t bo vi mu th l HS-3 cc nng khc nhau

323

Mu i chng m

Nng 5 g/ml

Nng 1 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml

Hnh IV.1.5.1.d: Hnh nh t bo vi mu th l HS-4 cc nng khc nhau

324

- Sau 72 gi

Mu i chng m

Nng 5 g/ml

Nng 1 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml

Hnh IV.1.5.1.e: Hnh nh t bo vi mu th l HS-3 cc nng khc nhau

325

Mu i chng m

Nng 5 g/ml

Nng 1 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml

Hnh IV.1.5.1.f: Hnh nh t bo vi mu th l HS-4 cc nng khc nhau

326

T cc hnh nh t bo chp c qua knh hin vi, chng ti c nhn xt

ban u nh sau:
- C 2 mu HS-3 v HS-4 u c kh nng c ch s phn chia v tng
trng ca t bo, c ch s to si (antimitotic activity) v to mng lin kt
gia cc t bo (anti-angiogenic activity) thng qua s thay i r rt v hnh thi
t bo, b mt mng t bo v kh nng to mng si bm dnh vo y phin
th nghim.
- Mu HS-3 c tc dng r rt hn biu hin s thay i v hnh thi
mng t bo v hnh nh pht trin ca t bo: sau 24 gi th nghim th nng
cht 5g/ml t bo gn nh khng pht trin v s lng, t bo bm dnh l
rt t. Sau 48 v 72 gi lng t bo bm dnh khng cn, t bo b bong ra
khi y phin th nghim v cht.
- Cc kt qu v hnh thi cng lm sng t hn cho kt qu th nghim
gy c t bo trn phin vi lng 96 ging sau 72 gi. Vi th nghim gy c
t bo, cc mu HS-3 v HS-4 u cho kt qu dng tnh mnh trn c 3 dng t
bo th.
Cc kt qu thu c (Th nghim gy c t bo v quan st hnh thi b
mt mng t bo) gi cho cc nghin cu tip theo mc cao hn mc
phn t mng t bo v tc dng gy ging ho- cht theo chng trnh
(apoptosis) c th lm r hn v c ch tc ng ca cc cht trn.
IV.1.5.2. Hot tnh khng vi sinh vt kim nh
Tt c cc hp cht phn lp c t loi hi sm Holothuria scabra
c th nghim v nh gi hot tnh khng vi sinh vt kim nh trn cc dng
vi sinh vt gy bnh tiu biu nht thuc cc nhm vi khun Gr(-); vi khun
Gr(+); nm mc v nm men. nh gi s b hot tnh, chng ti pha mu
nng theo tiu chun sng lc s b v tm cc mu th hin hot tnh khng
vi sinh vt kim nh nng . Cc mu c biu hin dng tnh bc s
b ny c pha long theo cc thang nng thp dn, t 5-10 thang nng
tnh gi tr nng ti thiu (MIC) m vi sinh vt b c ch pht trin gn
nh hon ton.
Kt qu nh gi hot tnh khng vi sinh vt kim nh cho thy: Ch c
hp cht HS1 v HS6 th hin hot tnh khng vi sinh vt kim nh. Mt iu
th v l cc hp cht ny khng rt mnh tt c cc chng nm vi gi tr MIC
u t mc thp l 50 g/ml, trong khi chng li khng c biu hin hot tnh
i vi cc loi vi sinh vt khc. iu ny th hin s ph hp hon ton vi cc
nghin cu cng b trn th gii (hin ch c 1 ti liu cng b v hot tnh
khng nm ca cc hp cht holothurin [240]).
Bng IV.1.5.2. Kt qu xc nh gi tr MIC ca HS1 v HS6
Hp cht

Vi khunGr(-)

Nng c ch ti thiu(MIC: g/ml)

Vi khunGr(+)
Nm mc

Nm men

E.coli

P.aeru

B.subti

S.aureus

Asp.niger

F.oxy

S.cere

C.albic

HS1

(-)

(-)

(-)

(-)

50

50

50

50

HS6

(-)

(-)

(-)

(-)

50

50

50

50

327

IV.1.6. Cc cng trnh khoa hc cng b

T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc
loi hi sm Holothuria scabra, chng ti bin son v ng ti 02 cng trnh
trn cc tp ch chuyn ngnh quc t v 01 bi trn Hi ngh quc gia:
(1) Nguyen Hai Dang, Nguyen Van Thanh, Phan Van Kiem, Le Mai Huong, Chau Van
Minh and Young Ho Kim, Two new triterpene glycosides from Vietnamese sea
cucumber. Archives of Phamarcal Research (Korea), 2007, 30(11), 1387-1391.
(2) Nguyen Van Thanh, Nguyen Hai Dang, Phan Van Kiem, Nguyen Xuan Cuong, Hoang
Thanh Huong, and Chau Van Minh, A new triterpene glycoside form the sea
cucumber Holothuria scabra collected in Vietnam. Asean Journal on Science and
Technology for Developtment, 2006, 23(4), 253-259.
(3) Nguyen Van Thanh, Chau Van Minh, Hoang Thanh Huong, Phan Van Kiem. Steroidal
constituents from Holothuria scabra living in Vietnamese sea. Tuyn tp cc bi bo
khoa hc Hi ngh Khoa hc ln th 20-HBK H Ni, phn ban Cng ngh Ha hu
c, 2006, 200-204.

Ngoi ra cng t cc kt qu ny, tp th tc gi xy dng v ng k

01 bng pht minh sng ch ti Cc S hu Tr tu Vit Nam l:
(1) - Chu Vn Minh, Phan Vn Kim, Nguyn Hi ng, Nguyn Vn Thanh, L Mai
Hng, Cng Thung, Trn Thu Hng, Nguyn Xun Cng. Hp cht Holothurin
A3 c hot tnh chng ung th v phng php chit hp cht ny t loi hi sm
Holothuria scabra, Bng c quyn sng ch s 6852, cp theo quyt nh s 2394/QSHTT, ngy 14 thng 02 nm 2008 ca Cc S hu Tr Tu Vit Nam

Cc kt qu nghin cu c tham gia gii thng khoa hc k thut

Trung ng on Thanh nin ln th XVIII - 2008 v ot gii khoa hc: s
1890/GTKHKTTN-2008 theo quyt nh s 1890/Q-KHCNVN ca Ch tch
Vin Khoa hc v Cng ngh Vit Nam ngy 18/11/2008 vi ti: Nghin cu
thnh phn ha hc, hot tnh sinh hc v ng dng ca loi hi sm trng Holothuria
scabra sinh sng ti cc vng bin Vit Nam.

Cc kt qu trn c s dng trong mt Lun n Tin s bo v

thnh cng (TS. Nguyn Vn Thanh)
Tng hp cc cht c phn lp t hi sm
Holothuria scabra
O
HO

21

O
HO

19

24

23

22
20
17

13

10

Xyl
4'

OH

Xyl

NaO3SO
6'' CH3

Qui
4''

OH

15

28

29

OH

MeGlc

2''

28

29

2'
O O
1''

OH

1''''

OCH3

OH

2''

O O

OH

CH2OH

4''''

Glc

4'''

6''''

OH

6
1'

CH2OH

2'

1''

OH

Qui
4''
6'''

O O

OH

1'''
2'''

OH

2''''

OH

HS1: Holothurin B

HS2: Holothurin A
328

15
30

O O

NaO3SO
6'' CH3

6
1'

13

5'

OH

30

O O

11

22
17

24

23

20

18
12

27

5'

10

25

4'

26
1

O
18

12

11

19

21

OH

26

25
27

21

26

OH

13

10

OH

NaO3SO

29

28

1'

OH

O O

CH2OH

4''''

OH

OH

2'

1''

CH2OH

2''

O O

6''''

CH2OH

OH

2'''

1''''

OCH3

OH

OH
1'''

MeGlc
4''''

2'''

OH

2''

Glc

4'''

1'''

1''''

OCH3

6
29

28

1'

OH

6'''

OH

4'''

15
30

Qui
4''

1''

OH

CH2OH

6''''

27

O O

O O
4''

OH

6'' CH3

6'' CH3

6'''

OH

O O

NaO3SO

2'

25

Xyl
4'

26

23

17

12

3
5'

O O

20

13

10

15
30

24

22

O
18

11

19

27

5'

O
HO

25

23

17

12

4'

20

18

11

19

24

HO

21

22

OH

2''''

2''''

OH

OH

HS4, Holothurin A3 (Cht mi)

HS3: Holothurin A2

OH

21
22

O
HO

27

OH

1'

19

2'

O O

1'''

4''

OH

OH

21
18

12
11

HO

OH

H
28

25

21
18

OH
27

30

29

HS7: 2,3,12, 21, 24R, 25hexahydroxylanosta-8- en

OH

27

15

(R)

(R)

H
OH

OH

OH

HS8: 24R- ergosta- 7,22dien- 3,5, 6- triol

329

25

14
8

HO

16

2
10

26

HO

13

19

NH 2

24
23

20
17

11

22

12

15

28

26

16

14

9
10

HS6, Holothurigenin B (Cht mi)

OH
24

23

20
17

HO

22

13

2''

OH

HS5, Holothurin A4 (Cht mi)

19

1''

OH

2'''

OH

OH

2'

6'' CH3

2''

4'''

OH

6
29

28

1'

OH

O
2''''

OH

OH

OCH3
OH

O O

1''

OH

O O
1''''

4''''

15
30

5'
4'

CH2OH

27

O
OH

13

O O
4''

10

29

6'' CH3

6'''

20
17

12

11

26
25

18

6
28

24

22

15
30

O O

NaO3SO

CH2OH

5'

6''''

23
21

OH

13

10

26
25

23

17

12

4'

20

18

11

19

24

HS9: Adenosine

IV.2. nghin cu ha hc theo nh hng hot tnh sinh

hc loi sao bin Archaster typicus (archasteridae)
IV.2.1. M u
Mu sao bin Archaster typicus c thu thp ti Ct B, Hi Phng vo
thng 1 nm 2006 v thng 8 nm 2007 v c TS. Cng Thung v cc cng
s, Vin Ti nguyn v Mi trng Bin, Vin KH&CN Vit Nam gim nh tn
khoa hc.
Sao bin l ng vt khng
xng sng, da gai. Thn
mm c dng sao nm
cnh (i khi nhiu cnh)
hoc hnh nm cnh, kch
thc t vi chc centimet
tr ln. C th dt bn,
nh dn v pha u cnh,
c nhng ng gn r t
gia c th ta ra v
ng vin chy ra
cnh. L ming v l hu
mn hai cc i din,
cc bn xng ca da c
gai nh.

Sao bin: Archaster typicus

Sao bin l ng vt bin n tht, n thch n loi c v nh trai, s, hn,...
Khi thiu thc n n n c ng loi, thm ch n c con chu mnh. Sao bin c
mt loi gn c th n c loi c v cng. Khi v c mt con s chng hn,
n ly chn m cht ly con mi cho n khi con s khng th chu c na, lc
2 mnh s m ra sao bin s nhn c hi tit dch tiu ha lm t lit con mi, th
d dy ra khi ming ri a vo khe gia 2 mnh v, t t tiu ha tht s. Khi
b cht hoc b k ch ngom chn, n t ng ct t chn chy.
Sao bin c kh nng ti sinh mnh, ch sau mt thi gian nhnh b trng thng
li mc di nh c.
Sao bin l mt hi sn qu, c tc dng nh hi sm, c dng lm
thuc bi dng c th cho nhng ngi yu mt, mi m dy, ph n mi sinh
v tr em chm ln. Cc nghin cu trn th gii cho thy, sao bin th hin
nhiu hot tnh th v v cha cc hot cht rt ng quan tm. Tuy nhin nc
ta hin nay c rt t cc cng trnh nghin cu v cc loi sao bin. Trong bo co
ny, chng ti trnh by qu trnh nghin cu ha hc theo nh hng hot tnh
sinh hc ca loi sao bin Archaster typicus. y l mt loi sao bin kh ph
bin nc ta tuy nhin hin c rt t cc cng trnh nghin cu v loi ny c v
ha hc v hot tnh sinh hc. Trn th gii, mi c 09 hp cht u l cc steroit
mang nhiu nhm hydroxyl c phn lp v xc nh cu trc t loi ny l:

330

CH2OH

CH2OH

OH

OH

OH

OH

OSO3H

OSO3H

HO

HO
OH

OH

OH

[241]

[241]
CH2OH

CH2OH
OH

OH
OH

OH
HO

OH

OH

OH
OH

OH

OSO3H

OH

OH

HO
OH

[241]

OH

[242]
OH

CH2OH

OH
OH

OH

OH

OH

OH

HO

OR

OH
OH

OH

OH

HO
OH

OH

OH

[241] R = H v R = SO3H

[242]
OH

OH

OH

OH

OH

OH

HO

OH

HO
OH
OH

OH
OH

OH

[241]

OH

[241]

IV.2.2. Phn lp cc hp cht

Sao bin Archaster typicus (5 kg kh) c ra sch, xay nh v chit
bng metanol (3 ln) bng thit b chit siu m. Dch metanol ca cc ln chit
c lc, gom li v ct loi dung mi nhit khng qu 60oC di p sut
gim thu c 200 g dch cn dch metanol. Cn dch ny c ha vo nc v
chit phn b ln lt vi cc dung mi clorofooc v butanol theo t l 1/1 (3 ln,
mi ln 6 lit) thu c cc cn dch tng ng l clorofooc 65 g, butanol 35 g v
nc 100 g (S IV.2.2.).
Cn chit clorofrom (65 g) c tin hnh chy ct th trn sc k ct pha
thng gradient nng axeton trong hexan thu c 4 phn on nh hn l
nhm 1 n 4. Cc phn on nhm 1, 2 v 3 c tip tc phn tch trn sc k
331

ct pha thng vi h dung mi ra gii thch hp thu c cc hp cht SB5,

sB6 v SB7 nh c ch ra trn s IV.2.2. Bng cc phng php tng t
cc hp cht SB1, SB2, SB3, v SB4 c phn lp t cn chit BuOH.
Sao bin Archaster
typicus (5 kg)
Thi nh, chit siu m trong
MeOH, ct loi dung mi
Dch chit MeOH
(200 g)
B sung nc
Chit CHCl3/H2O 1:1

B sung BuOH
Chit BuOH/H2O 1:1

Cn CHCl3
(65 g)

Cn BuOH
(35 g)

CC, silicagel, n-hexan/axeton

gradien phn cc tng dn

Nhm 8
CHCl3//MeOH/H2O 6/1/0,1
SB-4 (18 mg)

SB-3 (9 mg)
SB-2 (8 mg)

SB-1 (18 mg)

SB-7 (14 mg)

SB-6 (25 mg)

332

CC, Silicagel, YMC,

Axeton/MeOH/H2O
1/8/1

CHCl3//MeOH/H2O 8/1/0,1

n-hexan/axeton 5/1

Kt tinh trong
axeton

n-hexan/axeton 15/1
SB-5 (13 mg)

S IV.2.2. S phn lp cc hp cht t sao bin

Nhm 7

100% MeOH

Nhm 6

35% CHCl3

Nhm 5

70% CHCl3

Nhm 4

100% CHCl3

Nhm 3

CC, silicagel, CHCl3/MeOH gradien

phn cc tng dn
100% axeton

Nhm 2

35% n-hexan

70% n-hexan

100% nNhm 1

Cn pha nc
(100 g)

IV.2.3. Hng s vt l v d kin ph ca cc hp cht

IV.2.3.1. Hp cht SB1: 1-O-(-D-galactopyranosyl)-(2S,3S,4R)-2-[(2R)-2hydroxytetracosanoylamino]-14-methyl-pentadecane-1,3,4-triol
(Archastercerebroside - cht mi)
Cht bt v nh hnh mu trng, ph khi lng (ESI-MS) m/z: 818 [M+H]+,
ph khi lng phn gii cao HR-ESI-MS: m/z 840,6734 [M+ Na]+ (tnh ton l thuyt
cho cng thc C46H91NO10Na: 840,6541), cng thc phn t C46H91O10N (M = 817).
1
H-NMR (500 MHz, CDCl3+MeOD): (ppm): 3,58 (2H, dd, J = 5,5, 11,0 Hz,
H-1), 4,24 (1H, m, H-2), 4,11 (1H, dd, J = 6,0, 10,5 Hz, H-3), 3,83 (1H, m, H-4), 3,73
(1H, d, J = 5,5 Hz, H-2), 7,53 (1H, d, J = 9,2 Hz, NH), 0,87 (3H, t, J = 7,0 Hz, CH2CH2CH3), 0,85 (6H, d, J = 6,5 Hz, CH(CH3)2), 4,24 (1H, d, J = 7,5 Hz, H-1), 3,52
(1H, dd, J = 7,5, 8,0 Hz, H-2), 3,55 (1H, dd, J = 4,0, 8,0 Hz, H-3), 4,04 (1H, dd, J =
4,0, 8,0 Hz, H-4), 3,88 (1H, m, H-5), 3,50 (1H, dd, J = 5,5, 12,0 Hz, Ha-6), v 3,56
(1H, dd, J = 3,5, 12,0 Hz, Hb-6).
13
C-NMR (125 MHz, CDCl3+MeOD): (ppm): 69,18 (t, C-1), 50,81 (d, C-2),
74,82 (d, C-3), 72,66 (d, C-4), 176,14 (s, C-1), 72,55 (d, C-2), 14,15 (q, -CH2CH2CH3),
22,80 (q, CH(CH3)2), 103,96 (d, C-1), 71,57 (d, C-2), 73,79 (d, C-3), 69,66 (d, C-4),
75,38 (d, C-5), v 62,23 (t, C-6).
IV.2.3.2. Hp cht SB2: 27-Norcholestan-3,4,5,6,8,14,15,24R-octol.
Cht kt tinh mu trng, im chy 294oC, ph khi lng ESI-MS: m/z 509,2
[M+Na]+ (positive) v 485,4 [M-H]- (negative), cng thc phn t C26H46O8 (M = 486).
1
H-NMR (500 MHz, DMSO): (ppm): 1,11(1H, Ha-1), 1,41(1H, Hb-1), 1,35(1H,
Ha-2), 1,53(1H, Hb-2), 3,70(1H, H-3), 3,78(1H, H-4), 4,08 (1H, m, H-6), 1,66(1H, Ha7), 1,47(1H, Hb-7), 2,11 (1H, dd, J = 2,5, 13,0 Hz, H-9), 1,03(1H, Ha-11), 1,46(1H, Hb11), 1,38(1H, Ha-12), 1,62(1H, Hb-12), 4,29 (1H, dd, J = 4,0, 9,0 Hz, H-15), 1,64(1H,
Ha-16), 1,93 (1H, t, J = 12,0 Hz, Hb-16), 1,85 (1H, dd, J = 9,0, 18,0 Hz, H-17), 0,98
(3H, s, H-18), 1,06 (3H, s, H-19), 1,20 (1H, m, H-20), 0,73 (3H, J = 6,5 Hz, H-21), 0,90
(m); 1,42 (1H, H-22), 1,15(1H, Ha-23), 1,40(1H, Hb-23), 3,20 (1H, m, H-24), 1,22(1H,
Ha-25), 1,36(1H, Hb-25), 0,82 (3H, t, J = 7,0 Hz, H-26), 3,91 (1H, d, J = 4,0 Hz, 3-OH),
3,01 (1H, s, 6-OH), 3,58 (1H, s, 8-OH), 3,23 (1H, s, H-), v 4,15 (1H, 5,0).
13
C-NMR (125 MHz, DMSO): (ppm): 31,97 (t, C-1), 25,77 (t, C-2), 70,95 (d,
C-3), 66,94 (d, C-4), 76,77 (s, C-5), 64,40 (d, C-6), 38,99 (t, C-7), 77,26 (s, C-8), 39,49
(d, C-9), 38,53 (s, C-10), 16,89 (t, C-11), 33,95 (t, C-12), 46,70 (s, C-13), 83,06 (s, C14), 67,47 (d, C-15), 38,05 (t, C-16), 49,89 (d, C-17), 16,54 (q, C-18), 16,54 (q, C-19),
34,46 (d, C-20), 18,29 (q, C-21), 31,63 (t, C-22), 33,22 (t, C-23), 71,55 (d, C-24), 29,50
(t, C-25), v 9,89 (q, C-26).
IV.2.3.3. Hp cht SB3: 27-Norcholestan-3,4,5,6,7,8,14,15,24R-nonol
Cht kt tinh mu trng, im chy 288-290oC, ph khi lng ESI-MS ti m/z
501,3 [M-H]- (negative) v 525,2 [M+Na]+ (positive), cng thc phn t C26H46O9 (M =
502).
1
H-NMR (500 MHz, DMSO): (ppm): 1,16 (1H, Ha-1), 1,42 (1H, Hb-1), 1,38
(1H, Ha-2), 1,54 (1H, Hb-2), 3,78 (1H, H-3), 3,81 (1H, H-4), 3,80 (1H, d, J = 7,0 Hz, H-

333

6), 3,87 (1H, H-7), 2,18 (1H, dd, J = 2,5, 13,0 Hz, H-9), 1,13 (1H, Ha-11), 1,56 (1H, Hb11), 1,15 (1H, Ha-12), 1,38 (1H, Hb-12), 4,27 (1H, dd, J = 4,0, 9,0 Hz, H-15), 1,57 (1H,
Ha-16), 1,68 (1H, Hb-16), 1,82 (1H, dd, J = 9,0, 18,0 Hz, H-17), 1,03 (3H, s, H-18), 1,18
(3H, s, H-19), 1,26 (1H, H-20), 0,77 (3H, J = 6,5 Hz, H-21), 0,88 (1H, m, Ha-22), 1,45
(1H, Hb-22), 1,15 (1H, Ha-23), 1,38 (1H, Hb-23), 3,,24 (1H, m, H-24), 1,26 (1H, Ha-25),
1,37 (1H, Hb-25), 0,84 (3H, t, J = 7,0, H-26), 4,07 (1H, s, 3-OH), 3,98 (1H, d, J = 2,5
Hz, 4-OH), 3,43 (1H, s, 5-OH), 3,85 (1H, 6-OH), 3,53 (1H, s, 8-OH), 3,36 (1H, s, 14OH), v 4,18 (1H, d, J = 1,0 Hz, 24-OH).
13
C-NMR (125 MHz, DMSO): (ppm): 31,88 (t, C-1), 25,60 (t, C-2), 70,83 (d,
C-3), 69,13 (d, C-4), 76,94 (s, C-5), 66,62 (d, C-6), 71,83 (d, C-7), 78,41 (s, C-8), 39,49
(d, C-9), 38,35 (s, C-10), 16,85 (t, C-11), 33,92 (t, C-12), 47,33 (s, C-13), 82,39 (s, C14), 67,67 (d, C-15), 36,41 (t, C-16), 49,65 (d, C-17), 16,23 (q, C-18), 16,53 (q, C-19),
34,49 (d, C-20), 18,36 (q, C-21), 31,61 (t, C-22), 33,16 (t, C-23), 71,56 (d, C-24), 29,52
(t, C-25), v 9,86 (q, C-26).
IV.2.3.4. Hp cht SB4: Ergost-22-en-3,4,5,6,15,22E,24R,25R-nonol.
Cht kt tinh, tinh th mu trng, im chy 288-290oC, ph khi lng ESI-MS
xut hin pic ion phn t ti m/z 527,4 [M-H]+ (negative) v 551,3 [M+Na]+ (positive),
cng thc phn t C28H48O9 (M = 528).
1
H-NMR (500 MHz, DMSO): (ppm): 1,17 (1H, Ha-1), 1,42 (1H, Hb-1), 1,39
(1H, Ha-2), 1,56 (1H, m, Hb-2), 3,73 (1H, br t, J = 9,0 Hz, H-3), 4,30 (1H, H-4), 4,13
(1H, dd, J = 4,5, 11,5 Hz, H-6), 1,65 (1H, Ha-7), 1,47 (1H, br t, J = 12,5 Hz, Hb-7), 2,12
(1H, dd, J = 2,5, 13,0 Hz, H-9), 1,08 (1H, Ha-11), 1,50 (1H, Hb-11), 1,38 (1H, Ha-12),
1,67 (1H, Hb-12), 3,80 (1H, dd, J = 4,0, 9,0 Hz, H-15), 1,65 (1H, Ha-16), 1,89 (1H, t, J
= 12,0 Hz, Hb-16), 1,90 (1H, dd, J = 9,0, 18,0 Hz, H-17), 1,02 (3H, s, H-18), 1,18 (3H,
s, H-19), 2,12 (1H, m, H-20), 0,84 (3H, d, J = 6,0 Hz, H-21), 5,12 (1H, dd, J = 8,0, 15,5
Hz, H-22), 5,35 (1H, dd, J = 8,0, 15,5 Hz, H-23), 2,11 (1H, m, H-24), 3,21 (2H, dt, J =
5,0 17,0 Hz, H-26), 0,95 (3H, s, H-27), 0,84 (3H, t, J = 7,0 Hz, H-28), 3,05 (s, 5-OH),
3,62 (s, 8-OH), v 3,30 (s, 14-OH).
13
C-NMR (125 MHz, DMSO): (ppm): 31,98 (C-1), 25,77 (C-2), 70,95 (C-3),
67,51 (C-4), 76,78 (C-5), 64,40 (C-6), 38,05 (C-7), 77,28 (C-8), 39,49 (C-9), 38,50 (C10), 16,90 (C-11), 33,88 (C-12), 46,61 (C-13), 83,22 (C-14), 66,94 (C-15), 39,35 (C-16),
49,84 (C-17), 16,73 (C-18), 16,55 (C-19), 39,50 (C-20), 20,41 (C-21), 136,19 (C-22),
127,78 (C-23), 43,12 (C-24), 73,14 (C-25), 67,51 (C-26), 22,31 (C-27), v 15,05 (C-28).
IV.2.3.5. Hp cht SB5: cholesterol
Cht kt tinh mu trng, im chy 148-149 oC, C27H46O, M=386.
1
H-NMR (500 MHz, CDCl3) (ppm): 3,52 (1H, m, H-3), 5,35 (1H, br d, J = 5,0
Hz, H-6), 1,00 (3H, s, H-19), 0,91 (3H, s, H-21), 0,86 (6H, d, J = 6.5 Hz), H-26, 27) and
0.68 (3H, s, H-18).
13
C-NMR (125 MHz, CDCl3) (ppm): 37.2 (t, C-1), 31.6 (t, C-2), 71.8 (d, C-3),
42,2 (t, C-4), 140,7 (s, C-5), 121,7 (d, C-6), 31,8 (t, C-7), 31,8 (d, C-8), 50,1 (d, C-9),
36,5 (s, C-10), 21,1 (t, C-11), 39,8 (t, C-12), 42,3 (s, C-13), 56,7 (d, C-14), 24,3 (t, C15), 28,3 (t, C-16), 56,1 (d, C-17), 11,9 (q, C-18), 19,4 (q, C-19), 35,7 (d, C-20), 18,7 (q,

334

C-21), 36,1 (t, C-22), 23,8 (t, C-23), 39,5 (t, C-24), 28,0 (d, C-25), 22,5 (q, C-26), 22,8
(q, C-27).
IV.2.3.6. Hp cht SB6: cholest-7-en-3-one.
Tinh th mu trng, im chy 147-148oC, ph khi lng ESI-MS: m/z 385,2
[M+H]+ (C27H44O, M = 384).
1
H-NMR (500 MHz, CDCl3) (ppm): 1,47 (1H, m, Ha-1), 2,15 (1H, ddd, J =
4,5, 9,2, 13,6 Hz, Hb-1), 2,28 (1H, br d, J = 14,5 Hz, Ha-2), 2,42 (1H, dt, J = 6,0, 14,5
Hz, Hb-2), 5,18 (1H, br s, H-7), 0,57 (3H, s, H-18), 1,03 (3H, s, H-19), 1,39 (1H, m, H20), 0,93 (3H, d, J = 6,5 Hz, H-21), 0,86 (3H, d, J = 7,0 Hz, H-26), 0,87 (3H, d, J = 7,0
Hz, H-27).
13
C-NMR (125 MHz, CDCl3) (ppm): 38,8 (C-1), 38,1 (C-2), 211,8 (C-3), 44,2
(C-4), 42,9 (C-5), 30,1 (C-6), 116,9 (C-7), 139,5 (C-8), 48,9 (C-9), 34,4 (C-10), 21,7 (C11), 39,5 (C-12), 43,4 (C-13), 54,9 (C-14), 22,9 (C-15), 27,9 (C-16), 56,2 (C-17), 11,9
(C-18), 12,4 (C-19), 36,2 (C-20), 18,9 (C-21), 36,1 (C-22), 23,9 (C-23), 39,5 (C-24),
28,1 (C-25), 22,5 (C-26) v 22,8 (C-27).
IV.2.3.7. Hp cht SB7: batilol
Tinh th mu trng; im chy 70-71oC; ph khi lng ESI-MS m/z: 345,1
+
[M+H] , (C21H44O3, M = 344).
1
H-NMR (500 MHz, CDCl3) (ppm): 3,62 (1H, dd, J = 3,0 v 11,0 Hz, Ha-1),
3,71 (1H, dd, J = 5,0 v 11,0 Hz, Hb-1), 3,86 (1H, m, H-2), 3,52 (2H, m, H-3), 3,45 (2H,
m, H-1), 1,57 (2H, m, H-2), 1,26 (30H, br s, H-3 n H-17) v 0,88 (3H, t, J = 7,0
Hz, H-18).
13
C-NMR (125 MHz, CDCl3) :14,11 (q, C-18), 22,70 (t, C-17), 26,10 (t, C-3),
29,38-29,72 (13 CH2), 31,95 (t, C-16), 64,30 (t, C-3), 70,52 (d, C-2), 71,89 (t, C-1) v
72,50 (t, C-1).

335

IV.2.4. Xc nh cu trc ha hc ca cc hp cht

IV.2.4.1. Hp cht SB1: Archastercerebroside (cht mi)
Hp cht SB1 tch c t dch chit Butanol c dng bt trng. Cng
thc phn t c xc nh l C46H91O10N bng kt qu ph khi lng bn ph
nguyn t nhanh phn gii cao HR-FAB-MS cho pic ion m/z 840,6734 [M+ Na]+
positive (tng ng vi cng thc tnh ton l thuyt l C46H91NO10Na:
840,6541).

Hnh IV.2.4.1.a. Ph 1H-NMR ca hp cht SB1

Ph 1H-NMR ca SB1 c trng cho mt cerebroside vi cc tn hiu cng
hng ca 1 gc ng vi tn hiu proton anome ti 4,24 (1H, d, J = 7,5
Hz), 1 tn hiu proton c trng ca nhm -NH- ti 7,53 (J = 9,1 Hz) v tn
hiu ca cc proton metilen ca mch di ti 1,27 (36H, br s).
OH
OH OH
6"

4"

H
HO

(CH2)21CH3

H O

5"

3"

2'

1'

Gal

2"

OH
H

OH

NH

1"

(CH2)9CH(CH3)2

OH

Hnh IV.2.4.1.b. Cu trc ho hc ca SB1

336

Hnh IV.2.4.1.c. Ph 13C-NMR ca hp cht SB1

Hnh IV.2.4.1.d. Ph DEPT 90, 135 v 13C-NMR ca hp cht SB1

Ph 13C-NMR v DEPT cho tn hiu ca 1 nhm cacbonyl thuc lin kt
peptit C 176,14 ppm, 4 nhm cacbinol ti 69,28, 72,55, 72,66, 74,82 ppm, v 1
nhm metin ni vi nit 50,8 ppm. Cc tn hiu ca 1 gc ng cng c xc
nh 103,96 (cacbon anome); 62,23 ca nhm oxy metylen v cc tn hiu
trong khong 69,66 n 75,38 ca cc nhm oxi metin. vng trng mnh
l tn hiu ca 1 nhm metyl 14,14; 2 nhm metyl khc 39,36 ppm.

337

Hnh IV.2.4.1.e. Ph HSQC ca SB1

Bng IV.2.4.1: Kt qu ph NMR phn ceramide ca SB1
C

a, b

a, c

HMBC
(H to C)

Ceramide
1
2
3
4
1
2
NH
-CH2CH2CH3
-CH(CH3)2
Sugar
1
2
3
4
5
6

[243]
70,5
51,8
75,9
72,6
175,6
72,5

69,18
50,81
74,82
72,66
176,14
72,55

3,58 (dd, 5,5; 11,0)

4,24 (m)
4,11 (dd, 6,0; 10,5)
3,83 (m)
3,73 (dd, 5,5 Hz)
7,53 (d, 9,2 Hz)
0,87 (t, 7,0 Hz)
0,85 (d, 6,5 Hz)

C-1; C-2; C-3

C-1; C-3; C-4
C-1; C-2; C-4
C-2; C-3
-

4,24 (d, 7,5 Hz)

3,52 (dd, 7,5; 8,0)
3,55 (dd, 4,0; 8,0)
4,04 (dd, 4,0; 8,0)
3,88 (m)
3,50 (dd, 5,5; 12,0)
3,56 (dd, 3,5; 12,0)

C-1; C-3
C-1; C-3; C-4
C-1; C-2; C-5
C-2; C-3; C-5
C-3; C-6
C-4; C-5
C-4; C-5

14,3
22,8
[244]
104,5
71,7
73,8
69,7
76,0
62,0

14,15
22,80
103,96
71,57
73,79
69,66
75,38
62,23

o trong CDCl3+MeOD, b125MHz, c500MHz, *s liu phn ceramide ca Linkiacerebroside [243] v s

liu phn ng galactose ti liu [244]

338

Hnh IV.2.4.1.f. Ph HMBC ca SB1

OH
2'

1'

19

Gal
HO

5''

OH
4''

NH

6''

O
1''

OH

OH
4

3
8

1
2''

OH

OH

Hnh IV.2.4.1.g. Cc tng tc HMBC trong phn t SB1

T cc ph 2 chiu HSQC v HMBC chuyn dch ho hc ca C-H v
cc tng tc xa H-C c xc nh, kt qu trnh by trong bng 4.18. Kt
qu so snh cc gi tr ph (phn ceramide) ca SB1 vi Linkiacerebroside (mt
hp cht tch c t loi sao bin Linkia laevigata) bao gm (2S, 3S, 4R)
phytosphingozin v axit bo (2R)-2-hydroxy [243] hon ton ph hp. Da vo
ln ca hng s tng tc spin, cc v tr H-1 (J = 7,5 Hz); H-2 (J = 7,5;
8,0 Hz) v H-3(J = 4,0; 8,0 Hz) c xc nh l axial cn H-4(J = 4,0; 8,0
Hz) l equatorial, iu cho thy gc ng l galactoz. So snh cc gi tr
ph ca phn ng ca SB1 vi gc ng galactopyranosyl nhn thy cc v tr
339

hon ton trng khp [244]. Mt khc, lin kt gia gc ng vi v tr C-1 cng
c xc nhn thng qua tng tc HMBC gia H-1 vi C-1. Ngoi ra trt t ca
cc v tr C-1, C-2, C-3, C-4, C-1, C-2 cng c khng nh bi cc tng tc
ca H-1 vi C-2; C-3; gia H-2 vi C-1, C-3, C-4, C-1, gia H-3 vi C-1, C-2, C4, gia H-2, vi C-1, gia H ca nhm NH vi C-1, C-2.

Hnh IV.2.4.1.h. Ph khi lng ESI-MS ca phn nhnh axit bo ca SB1

xc nh cu trc ho hc ca cc mch di, chng ti tin hnh
phn ng thu phn SB1 bng hn hp HCl 5% trong MeOH 700C trong 20 gi.
Sau hn hp phn ng c chit bng n-hexan, phn axit bo thu phn ra s
nm trong lp hexan. Lp MeOH c trung ho bng dung dch Ag2CO3, lc b
kt ta ri c cn, thu c hn hp sn phm gm ng v ancol sphingozine
mch di.
Vi cng thc phn t xc inh c phn trn, SB1 c khng no l
2, tng ng vi 1 nhm cacbonyl v 1 vng pyrano ca 1 gc ng, do cc
mch di trong phn t u l mch cacbon no. Do trong phn t SB1 tn ti 3
nhm methyl nn trong 2 mch di ny s c 1 mch thng v 1 mch nhnh.
Da vo cc tng tc HMBC chng ti xc nh c mch nhnh s c dng
CH(CH3)CH3. Mt khc t kt qu ph khi lng EI-MS c th nhn thy axit
bo c dng mch thng bi cc pic ion m/z 383,4; M/z 369,4; m/z 355,4; m/z
341,4 lun hn km nhau 14 n v. Vi pic ion phn t m/z 383,4 [M-H]+
(negative) cho php xc nh cng thc hydroxy axit bo l CH3 (CH2)21CHOHCOOH. T d dng xc nh c cng thc gc ancol sphyngosine v SB1 s
l
1-O-(-D-galactopyranosyl)-(2S,3S,4R)-2-[(2R)-2-hydroxytetracosanoylamino]-14-methyl-pentadecane-1,3,4-triol. y l mt hp cht mi v c
chng ti t tn l archastercerebroside.

340

Hnh IV.2.4.1.i. Ph khi lng phn gii cao HR-FAB-MS ca SB1

Hnh IV.2.4.1.j. Ph khi lng ESI-MS ca SB1

341

IV.2.4.2. Hp cht SB2: 27-Norcholestan-3,4,5,6,8,14,15,24R-octol.

Hp cht SB2 c phn lp t dch chit BuOH, dng tinh th, mu
trng. Cng thc phn t ca n c xc nh l C26H46O8 (M = 486) da trn kt
qu ph khi lng ESI-MS [ti m/z 509,2 [M+Na]+ (positive) v 485,4 [M-H](negative)] v ph cng hng t ht nhn.

Hnh IV.2.4.2.a. Ph 1H-NMR ca hp cht SB2

OH
21

22
20

18
12
11

19
1

HO

10

25

26

23

17

13

OH

24

16
14

OH

15

OH

OH
OH

OH

Hnh IV.2.4.2.b. Cu trc ha hc ca SB2

Ph 1H-NMR ca SB2 c trng cho mt hp cht steroit. Cc tn hiu
cng hng trn ph ny nm hai vng rt r rt: nm vng trng thp l cc
tn hiu proton ca cc nhm OH v oximetin, nm vng trng cao l tn hiu
proton ca cc nhm metin, metilen ca mch nhnh v khung steroit, ngoi ra
cc tn hiu cng hng ti 0,98 (3H, s, H-18), 1,06 (3H, s, H-19), 0,73 (3H, J
= 6,5 Hz, H-21), v 0,82 (3H, t, J = 7,0 Hz, H-26) khng nh tng ng cho s
tn ti ca 2 nhm metyl bc ba v 2 nhm metyl bc hai.

342

Hnh IV.2.4.2.c. Ph 13C-NMR ca SB2

Ph 13C-NMR xut hin tn hiu cng hng ca 26 cacbon. Trong , s
tn ti ca 5 nhm oximetin v 3 cacbon bc bn ni vi nguyn t xy c
khng nh tng ng bi cc tn hiu cng hng ti 70,95 (d, C-3), 66,94 (d,
C-4), 64,40 (d, C-6), 67,47 (d, C-15), 71,55 (d, C-24), 76,77 (s, C-5), 77,26 (s, C8), v 83,06 (s, C-14). Cc tn hiu ti 16,54 (q, C-18), 16,54 (q, C-19), 18,29 (q,
C-21), v 9,89 (q, C-26) khng nh s c mt ca 4 nhm metyl.

Hnh IV.2.4.2.d. Ph 13C-NMR v cc ph DEPT ca SB2

343

Bng IV.2.4.2. Kt qu ph NMR ca SB2

C

Ca,b

Ha,c (J, Hz)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
3-OH
6-OH
8-OH
14-OH
24-OH

31,97 (t)
25,77 (t)
70,95 (d)
66,94 (d)
76,77 (s)
64,40 (d)
38,99 (t)
77,26 (s)
39,49 (d)
38,53 (s)
16,89 (t)
33,95 (t)
46,70 (s)
83,06 (s)
67,47 (d)
38,05 (t)
49,89 (d)
16,54 (q)
16,54 (q)
34,46 (d)
18,29 (q)
31,63 (t)
33,22 (t)
71,55 (d)
29,50 (t)
9,89 (q)

1,11*; 1,41*
1,35*, 1,53*
3,70*
3,78*
4,08 (1H, m)
1,66*, 1,47*
2,11 (1H, dd, 2,5, 13,0)
1,03*; 1,46*
1,38*, 1,62*

4,29 (1H, dd, 4,0, 9,0)

1,64*; 1,93 (t, 12,0)
1,85 (1H, dd, 9,0, 18,0)
0,98 (3H, s)
1,06 (3H, s)
1,20 (m)
0,73 (3H, 6,5 Hz)
0,90 (m); 1,42 *
1,15*; 1,40*
3,20 (1H, m)
1,22*; 1,36*
0,82 (3H, t, 7,0)
3,91 (1H, d, 4,0)
3,01 (1H, s)
3,58 (1H, s)
3,23 (1H, s)
4,15 (1H, 5,0)

o trong DMSO, b125MHz, c500MHz, *tn hiu b che khut

344

HMBC
(C to H)
H-19

H-6
H-19, 6-OH
6-OH
6-OH
8-OH
H-19, 8-OH
H-19
H-18
H-18, 14-OH
H-18, 8-OH, 14-OH
14-OH
H-18, H-21
H-17
H-21
H-17
H-17, H-21
24-OH
24-OH, H-26
24-OH

Hnh IV.2.4.2.e. Ph HSQC ca SB2

Hnh IV.2.4.2.f. Ph HMBC ca SB2

345

OH

OH
OH

OH

HO
OH
OH

OH

Hnh IV.2.4.2.g. Cc tng tc HMBC chnh ca SB2

T cc phn tch nu trn c th s b xc nh, SB2 l mt steroit c 26
cacbon, cha 8 nhm OH trong phn t v ch c 4 nhm metyl. Thng qua ph
HSQC, s liu ph cng hng t ht nhn ca cc cacbon c gn vi s liu
ph ca cc proton tng ng (bng IV.2.4.2).

Hnh IV.2.4.2.h. Ph ROESY ca SB2

Phn tch chi tit cc tng tc xa (C-H) trn ph HMBC (bng IV.2.4.2
v hnh IV.2.4.2.g) cho php xc nh v tr ca cc nhm th, khu ni cc mnh
cu trc li vi nhau v xy dng nn cu trc ton phn ca SB2 nh c ch ra
hnh IV.2.4.2.b. Ngoi ra, cu hnh 4,6,8,15 ca cc nhm th hydroxy

346

c xc nh nh cc tng tc H-19 vi OH-4 v H-6; H-18 vi OH-8 v H-15

nhn c trn ph cng hng t ht nhn hai chiu tng tc khng gian
ROESY (hnh IV.2.4.2.h). Tng t cc tng tc gia H-17 vi OH-14 v OH-6
vi H-3 gii thch v tr OH-14 c cu hnh v OH-3 l . T tt c cc phn
tch nu trn, cu trc ha hc ca SB2 c xc nh l 27-norcholestan3,4,5,6,8,14,15,24R-octol. Trn th gii, hp cht ny c phn lp
t loi sao bin Archaster typicus [241]. Tuy nhin, y l ln u tin hp cht
ny c cng b t loi A. typicus sinh sng ti cc vng bin Vit Nam.

Hnh IV.2.4.2.i. Ph khi lng ESI-MS ca SB2

347

IV.2.4.3. Hp cht SB3: 27-Norcholestan-3,4,5,6,7,8,14,15,24Rnonol

Hp cht SB3 c phn lp di dng tinh th mu trng. Thng qua kt
qu ph khi lng ESI-MS ti m/z 501,3 [M-H]- (negative) v 525,2 [M+Na]+
(positive) v ph cng hng t ht nhn cho php xc nh cng thc phn t
ca n l C26H46O9 (M = 502).

Hnh IV.2.4.3.a. Ph 1H-NMR ca SB3

OH
21

22
20

18
12
11

19
1

HO

10

26

23

16
14

15

8
7

OH

OH

OH

OH
OH

25

17

13

OH

24

OH

Hnh IV.2.4.3.b. Cu trc ha hc ca SB3

Ph 1H-NMR ca SB3 cng c trng cho mt hp cht steroit v c cc
tn hiu cng hng nm hai vng r rt, mt vng l cc tn hiu proton ca
cc nhm OH v oximetin v mt vng l tn hiu proton ca cc nhm metin v
metilen ca khung steroit v mch nhnh. S tn ti ca 2 nhm metyl bc ba v 2
nhm metyl bc hai c khng nh bi cc tn hiu cng hng ti 1,03 (3H,

348

s, H-18), 1,18 (3H, s, H-19), 0,77 (3H, J = 6,5 Hz, H-21), v 0,84 (3H, t, J = 7,0,
H-26).

Hnh IV.2.4.3.c. Ph 13C-NMR ca SB3

Hnh IV.2.4.3.d. Ph 13C-NMR v cc ph DEPTca SB3

349

Bng IV.2.4.3. Kt qu ph NMR ca SB3

C

a,b

a,c (J, Hz)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
3-OH
4-OH
5-OH
6-OH
8-OH
14-OH
24-OH

31,88 (t)
25,60 (t)
70,83 (d)
69,13 (d)
76,94 (s)
66,62 (d)
71,83 (d)
78,41 (s)
39,49 (d)
38,35 (s)
16,85 (t)
33,92 (t)
47,33 (s)
82,39 (s)
67,67 (d)
36,41 (t)
49,65 (d)
16,23 (q)
16,53 (q)
34,49 (d)
18,36 (q)
31,61 (t)
33,16 (t)
71,56 (d)
29,52 (t)
9,86 (q)

1,16 *; 1,42 *
1,38*; 1,54*
3,78 *
3,81*
3,80 (1H, d, 7,0)
3,87*
2,18 (1H, dd, 2,5, 13,0)
1,13*; 1,56*
1,15*; 1,38*

4,27 (1H, dd, 4,0, 9,0)

1,57*; 1,68*
1,82 (1H, dd, 9,0, 18,0)
1,03 (3H, s)
1,18 (3H, s)
1,26 *
0,77 (3H, 6,5 Hz)
0,88 (m); 1,45 *
1,15*; 1,38*
3,,24 (1H, m)
1,26*; 1,37*
0,84 (3H, t, 7,0)
4,07 (s)
3,98 (d, 2,5 Hz)
3,43 (s)
3,85*
3,53 (s)
3,36 (s)
4,18 (d, 1,0)

HMBC
(C H)
H-19
H-4
H-3, 5-OH, 4-OH
H-19, 5-OH
6-OH
8-OH, H-6
8-OH
H-19, 8-OH
H-9, H-19, 5-OH
H-9
H-11, H-18
H-18, 14-OH
H-18, 14-OH
14-OH
H-20
H-18, H-21
H-1
H-17, H-21
H-17
H-21
24-OH
H-26, 24-OH
H-26, 24-OH

o trong DMSO, b125MHz, c500MHz, *tn hiu b che khut

Ph 13C-NMR ca SB3 cng sut hin cc tn hiu cng hng ca 26

cacbon. S liu ph 1H v ca SB3 rt tng ng vi s liu ca SB2. S khc
bit nhn thy nht trn ph 13C-NMR gia hai hp cht ny l s xut hin
thm tn hiu cng hng ca mt nhm oximetin ti 71,83 (d, C-7) v s mt
i tn hiu cng hng ca mt nhm metilen vng trng cao ca SB3 so vi
SB2. Cng vi kt qu ph khi lng cho php khng nh cu trc ha hc ca

350

SB3 hon ton tng t nh ca SB2 ch khc s xut hin thm mt nhm OH
trong phn t.

Hnh IV.2.4.3.e. Ph HSQC ca SB3

Hnh IV.2.4.3.f. Ph HMBC ca SB3

351

OH
21

22

26

20

24

18
12
11

19

HO

10

16
14

15

8
7

OH

OH

OH

OH
OH

25

17

13

OH

23

OH

Hnh IV.2.4.3.g. Cc tng tc HMBC chnh ca SB3

Phn tch chi tit cc tng tc trn ph HSQC v HMBC cho php xc
nh v tr ca cc nhm th cng nh cu trc ton b phn t ca SB3. Nhm
OH xut hin thm c khng nh l gn vo v tr C-7 thng qua cc tng tc
HMBC gia proton ca nhm OH ti C-8 [H 3,53 (1H, s)] v proton H-6 [H 3,80
(1H, d, J = 7,0 Hz)] vi cacbon C-7 [C 71,83 (d)]. T tt c cc phn tch nu
trn, cu trc ha hc ca SB3 c xc nh l 27-Norcholestan3,4,5,6,7,8,14,15,24R-nonol. Hp cht ny cng c nhm nghin
cu ca gio s Riccio phn lp v xc nh cu trc t loi sao bin Archaster
typicus [241]. Tuy nhin, y l ln u tin n c cng b t loi sao bin ny
sinh sng ti cc vng bin ca Vit Nam.

Hnh IV.2.4.3.h. Ph khi lng ESI-MS ca SB3

352

IV.2.4.4. Hp cht SB4: Ergost-22-en-3,4,5,6,15,22E,24R,25R-nonol.

Hp cht SB4 c phn lp di dng tinh th mu trng. Ph khi lng
ESI-MS xut hin pic ion phn t ti m/z 527,4 [M-H]+ (negative) v 551,3
[M+Na]+ (positive), cng vi kt qu ph cng hng t ht nhn cho php xc
nh cng thc phn t ca n l C28H48O9 (M = 528).

Hnh IV.2.4.4.a. Ph 1H-NMR ca SB4

28
21

22
20

18
12
11

19
1

HO

10

25

23

CH2OH
OH

27

17

13

OH

26
24

16
14

15

OH

OH

OH
OH

OH

Hnh IV.2.4.4.b. Cu trc ha hc ca SB4

Tng t nh hp cht SB2 v SB3, ph 1H-NMR ca SB4 cng xut hin
cc tn hiu c trng ca mt hp cht steroit cha nhiu nhm OH. S khc bit
d nhn thy nht trn ph 1H-NMR ca hp cht ny so vi hai hp cht kia l:
s xut hin thm tn hiu cng hng ca mt ni i ngoi vng c cu hnh
trans ti 5,12 (1H, dd, J = 8,0, 15,5 Hz, H-22)/5,35 (1H, dd, J = 8,0, 15,5 Hz,
H-23), v s xut hin tn hiu ca 5 nhm metyl trong c 3 nhm metyl bc
353

ba ti 1,02 (3H, s, H-18), 1,18 (3H, s, H-19), v 0,95 (3H, s, H-27), v 2 nhm
metyl bc hai ti 0,84 (6H, d, J = 6,0 Hz, H-21 v H-28).

Hnh IV.2.4.4.c. Ph 13C-NMR ca SB4

Hnh IV.2.4.4.d. Ph 13C-NMR v cc ph DEPT ca SB4

354

Bng IV.2.4.4. Kt qu ph NMR ca SB4

C

a,b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
3-OH
5-OH
6-OH
8-OH
14-OH

31,98
25,77
70,95
67,51
76,78
64,40
38,05
77,28
39,49
38,50
16,90
33,88
46,61
83,22
66,94
39,35
49,84
16,73
16,55
39,50
20,41
136,19
127,78
43,12
73,14
67,51
22,31
15,05

a,c
(J, Hz)
1,17*-1,42*
1,39*-1,56 (m)
3,73 (1H, br t, 9,0)
4,30*
4,13 (dd, 4,5; 11,5)
1,65*-1,47 (br t, 12,5)
2,12 (1H, dd, 2,5; 13,0)
1,08*-1,50*
1,38*-1,67*

3,80 (1H, dd, 4,0; 9,0)

1,65*-1,89 (t, 12,0)
1,90 (1H, dd, 9,0; 18,0)
1,02 (3H, s)
1,18 (3H, s)
2,12 (m)
0,84 (3H, d, 6,0)
5,12 (1H, dd, 8,0; 15,5)
5,35 (1H, dd, 8,0; 15,5)
2,11 (1H, m)
3,21 (2H, dt, 5,0; 17,0)
0,95 (3H, s)
0,84 (3H, d, 7,0)

HMBC
(CH)
H-19
H-3

ROESY

5-OH
14-OH
H-3; H-19; 5-OH
5-OH
8-OH
8-OH
8-OH; H-19
5-OH; H-19

H-19

H-18
H-18; 14-OH
H-18; 14-OH
14-OH
H-18; H-21
8-OH
8-OH
H-21; H-23
H-22
H-21; H-25
H-20; H-28
H-28; H-22; H-27
H-27; H-28
H-24; H-27
H-24
H-24

3,05 (s)

H-3

3,62 (s)
3,30 (s)

H-18; H-19
H-4; 14-OH

o trong DMSO, b125MHz, c500MHz, *tn hiu b che khut

Ph 13C-NMR ca SB4 xut hin tn hiu cng hng ca 28 cacbon.

Trong , mt ni i ngoi vng c cu hnh trans c khng nh bi cc tn
hiu cng hng ti 136,19 (CH, C-22)/127,78 (CH, C-23), 3 nhm metyl bc
ba v 2 nhm metyl bc hai c khng nh tng ng bi cc tn hiu cng
hng ti 16,73 (C-18), 16,55 (C-19), 22,31 (C-27), 20,41 (C-21), v 15,05 (C28). Ngoi ra, cc tn hiu ti 70,95 (CH, C-3), 67,51 (CH, C-4), 64,40 (CH, C6), 66,94 (CH, C-15), 76,78 (C, C-5), 77,28 (C, C-8), 83,22 (C, C-14), v 67,51
(CH2, C-26), khng nh tng ng cho s tn ti ca 4 nhm oximetin, 4 cacbon
bc bn ni vi nguyn t xy v 1 nhm oximetilen.
355

Hnh IV.2.4.4.e. Ph HSQC ca SB4

Hnh IV.2.4.4.f. Ph HMBC ca SB4

356

S liu ph 1H v 13C-NMR ca phn khung steroit ca SB4 gn nh trng

khp hon ton vi s liu ti cc v tr tng ng ca SB2 cho php khng nh
hai hp cht ny c cng cu trc phn khung, ch khc nhau cu trc ca mch
nhnh.
CH2OH
OH
OH
OH

OH
HMBC

HO
OH
OH

COSY

OH

Hnh IV.2.4.4.g. Cc tng tc COSY v HMBC chnh ca SB4

Hnh IV.2.4.4.h. Ph H-H COSY ca SB4

Phn tch chi tit cc tng tc trn ph H-H COSY cho php ghp ni cc
mnh cu trc t C-1 n C-4, C-6 sang C-7, C-9 sang C-11 sang C-12, C-15 n
C-17, C-17 sang C-20, v C-20 n C-24 (hnh IV.2.4.4.g). Kt qu ny, cng vi
cc tng tc HMBC ca H-28 sang C-25, H-24 sang C-26, H-27 sang C-24/C-

357

25/C-26 cho php khng nh chnh xc cu trc ca nhnh bn nh c ch ra

trn hnh IV.2.4.4.b.

Hnh IV.2.4.4.i. Ph ROESY ca SB4

Hnh IV.2.4.4.j. Ph ESI-MS ca SB4

358

Cu hnh ca cc nhm OH v metyl c xc nh thng qua cc tng

tc khng gian trn ph ROESY. T tt c cc phn tch nu trn, cu trc ha
hc ca SB4 c xc nh l ergost-22-en-3,4,5,6,15,22E,24R,25R-nonol.
Nm 1989, nhm nghin cu ca gio s Riccio cng b s phn lp v xc
nh cu trc ca hp cht ny t loi sao bin Archaster typicus [242]. Tuy
nhin, y l ln u tin hp cht ny c thng bo t loi sao bin A. typicus
sinh sng ti cc vng bin ca Vit Nam.
IV.2.4.5. Hp cht SB5: cholesterol
Hp cht SB5 nhn c di dng tinh th hnh kim c mu trng. Th
nghim trn sc k lp mng (TLC) vi thuc th c hiu (phun dung dch
H2SO4 10% ri h nng t t cho mu tm hng, sau chuyn dn sang tm
xanh) cho thy y l mt steroit. Hp cht ny thu c vi hm lng rt cao
cng vi gi tr rf = 0,35 trn TLC vi h dung mi l Hexan/Axeton 7/1 v gi
tr nhit nng chy (mp 148,5oC) ca n gi ti hp cht cholesterol, mt
sterol ph bin nht trong gii ng vt. Cng vi s ph hp hon ton v s liu
ph 1H- v 13C-NMR (xem phn 2.5.5) ca SB5 so vi ti liu cng b [220]
cho php khng nh hp cht ny chnh l cholesterol.
21

26
20

18

11

19

14

15

10
3

27

17

13

25

22

7
6

HO

Hnh IV.2.4.5.a. Cu trc ha hc ca SB5

IV.2.4.6. Hp cht SB6: cholest-7-en-3-one
Hp cht SB6 phn lp c di dng tinh th mu trng. Ph 1H- v 13CNMR ca SB6 tng t nh ca SB5 cho thy hai hp cht ny c cu trc tng
i ging nhau. S trng khp hon ton v gi tr ph 13C-NMR ti cc v tr
tng ng t C-11 n C-27 cho php khng nh phn cu trc ny ca hai hp
cht l ging nhau.
21
25
18

17

11

9
10

27

13

19
1

20

14
8

Hnh IV.2.4.6.a. Cu trc ha hc ca SB6

359

26

Hnh IV.2.4.6.b . Ph 1H-NMR ca SB6

Hnh IV.2.4.6.c. Ph 13C-NMR ca SB6

Trn ph 1H-MNR ca SB6 xut hin cc tn hiu c trng ca mt
proton olefin ti 5,18 (1H, s), nm nhm metyl ti 0,57 (3H, s), 1,03 (3H, s),
0,93 (3H, d, J = 6,5 Hz), 0,86 (3H, d, J = 7,0 Hz) v 0,87 (3H, d, J = 7,0 Hz). S
khc bit d nhn thy nht trn ph 13C-NMR gia hai hp cht ny l tn hiu
cng hng ca nhm oximetin C-3 ti 71,8 (d) b mt v thay vo l s
360

xut hin ca nhm cacbonyl ti 211,81. Ngoi ra, gi tr chuyn dch ha

hc ca ni i thay i t 140,7 (s, C-5)/121,7 (d, C-6) sang 116,97
(d)/139,56 (s) cho thy v tr ni i b thay i. V tr ni i ti C-7/C-8 v
nhm cacbonyl ti v tr C-3 c xc nh bi s trng khp hon v cc s liu
phn khung steroit ca SB6 vi hp cht c cu trc tng t [245].

Hnh IV.2.4.6.d. Ph 13C-NMR v cc ph DEPT ca SB6

Hnh IV.2.4.6.e. Ph HSQC ca SB6

361

Hnh IV.2.4.6.f. Ph HMBC ca SB6

Hnh IV.2.4.6.g. Ph khi lng (ESI-MS) ca SB6

Ngoi ra, phn tch chi tit cc tng tc trn ph HMBC mt ln na
khng nh v tr ca lin kt i nh nu phn trn. T cc tt c phn tch
nu trn cng vi kt qu ph khi lng ESI-MS ti m/z 385,2 [M+H]+ cho php
khng nh cu trc ha hc ca 3 l cholest-7-en-3-one.
362

IV.2.4.7. Hp cht SB7: Batyl alcohol

Hp cht SB7 nhn c di dng kt tinh mu trng, ph khi lng
(ESI-MS) m/z: 345 [M+H]+.

Hnh IV.2.4.7.a. Ph 1H-NMR ca SB7

3

HO

OH

1'

18'

CH2(CH2)14CH3

Hnh IV.2.4.7.b. Cu trc ha hc ca SB7

Trn ph 1H-NMR ca SB7 xut hin cc tn hiu c trng ca cc nhm
oximetin v oximetilen ti 3,86 (1H, m), 3,71(1H, dd, J = 5,0, 11,0 Hz), 3,64
(1H, dd, J = 3,0, 11,0 Hz), 3,52 (2H, m) v 3,45 (2H, m); cc proton ca cc
nhm OH ti 2,64 (1H, br s) v 2,17 (1H, br s); mt nhm metyl ti 0,88 (3H,
s) v cc nhm metilen ti 1,57 (2H, m) v 1,26 (28H, br s).

363

Hnh IV.2.4.7.c. Ph 13C-NMR ca SB7

Trn ph 13C-NMR xut hin cc tn hiu c trng ca mt mch di
cacbon no ti 31,95 (t), 29,72 (t), 29,68 (t), 29,61 (t), 29,48 (t), 29,38 (t), 26,10
(t), 22,70 (t) v 14,12 (q). Cc tn hiu ca mt gc glyxerin b th mt v tr v
khng i xng -CH2OCH2CH(OH)CH2OH cng c khng nh bi s xut
hin cc tn hiu cacbon ti 72,51 (t, C-1), 70,52 (d, C-2), 64,30 (t, C-3) v
71,89 (t, C-1) [221]. Theo cc ti liu cng b [221[219]222], tt c cc cc
te glyxerit phn lp c cho n nay u dng cu hnh (S) cho php d
on cu hnh ti v tr C-2 l dng (S).
Bng IV.2.4.7. Kt qu ph NMR ca SB7.
C

C [221]

71,8

2
3
1
2
3
4-15
16
17
18
2-OH
3-OH

70,5
64,2
72,4
29,0
26,1
29,3-29,7
31,9
22,7
14,1
-

C a, b

Ha, c (J, Hz)

72,51 (t)
70,52 (d)
64,30 (t)
71,89 (t)
2,9,30-29,72 (t)
26,11 (t)
29,30 - 29,72 (t)
31,95 (t)
22,71 (t)
14,12 (q)
-

o trong CDCl3, b125MHz, c 500MHz, *gi tr c th i ch cho nhau.

364

3,64 (dd, 3,0, 11,0)

3,71 (dd, 5,0, 11,0)
3,86 (m)
3,52 (m)
3,45 (m)
1,58 (m)
1,26 (br s)
1,26 (br s)
1,26 (br s)
1,26 (br s)
0,88 (t, 7,0)
2,17 * (br s)
2,64 * (br s)

Hnh IV.2.4.7.d. Ph 13C-NMR v cc ph DEPT ca SB7

Cc phn tch nu cho php khng nh SB7 l mt glyxerol mono
alkyl te. T , s liu ph NMR ca n c so snh vi cc s liu c
cng b ca batyl alcohol [221]. S ph hp hon ton ti cc gi tr ph tng
ng cng vi s xut hin pc ion phn t ti m/z 345 trn ph khi lng ESI-MS
(tng ng vi cng thc phn t C21H44O3, M=344) cho php xc nh chnh xc
di ca mch cc bon no v cu trc ha hc ca SB7 l chimyl alcohol
(glycerol 1-O-octadecyl ether). iu ny mt ln na c khng nh bi cc
tng tc trn ph cng hng t ht nhn hai chiu tng tc xa HMBC.

Hnh IV.2.4.7.e. Ph HSQC ca SB7

365

OH

HO

2
3

1'

18'

CH2-(CH2)16-CH3

Hnh IV.2.4.7.f. Ph HMBC ca SB7

Hnh IV.2.4.7.g. Ph khi lng ESI-MS ca SB7

366

IV.2.5. nh gi hot tnh sinh hc ca cc cht

IV.2.5.1. Kt qu nh gi hot tnh khng NF-B ca cc hp cht
Cc hp cht phn lp c t loi sao bin Archaster typicus c tin
hnh th nghim hot tnh khng NF-B, kt qu th nghim c th c nu ra
trong bng IV.2.5.1.
NF-B l mt yu t phin m c tm quan trng c bit trong cc p
ng min dch v vim. Yu t ny ln u tin c nhn dng l mt cht iu
tit s biu hin ca gen chui nh Kappa trong cc t bo lympho B chut, v
sau c tm thy nhiu t bo khc nhau. Nhiu protein NF-B khc nhau
c xc nh c tnh. Dng c hot ha ca NF-B l mt d ime, thng
gm 2 protein: mt tiu n v p65 (cng c gi l relA) v mt tiu n v
p50. Cc tiu n v khc nh rel, relB, v-rel cng c th l thnh phn ca NFB c hot ha v c l l cc dng khc nhau ca NF-B c th hot ha cc
tp hp khc nhau ca gen dch. Trong cc t bo khng c kch thch, NF-B
c tm thy trong bo tng v n b lin kt i IB v IB m cc phn t
ny ngn cn NF-B i vo nhn t bo. Khi cc t bo v kch thch th cc
enzym kinase c hiu phosphoryl ha IB lm cho n v gim phn nhanh trong
bi cc tp hp enzym protease. S gii phng NF-B t IB dn ti s chuyn
NF-B vo nhn t bo, ti y n lin kt vi cc on trnh t c hiu trong
vng khi u phin m ca cc gen ch.
Bng IV.2.5.1. Kt qu th nghim hot tnh khng NF-B
Hp cht

IC50 (M)

Parthelolide
(chng dng)
SB1
SB2
SB3
SB4
SB5
SB6
SB7

6.66 0.07
34,71,84
30,82,12
28,11,98
27,81,87
>50
>50
>50

NF-B iu tit s biu hin ca nhiu gen tham gia vo cc p ng min

dch v vim. N lm tng s biu hin ca cc gen quy nh nhiu cytokine, cc
enzym v cc phn t dnh kt trong cc bnh vim mn tnh. NF-B c hot
ha bi nhiu yu t khc nhau (virus, cc cht xy ha v cc khng nguyn)
lm tng p ng vim. S hot ha ny li dn ti s biu hin phi hp ca
nhiu gen m ha cc protein (nh cc cytokine, chemokine, enzym v cc phn
t dnh kt) tham gia vo s tng hp cht trung gian v ko di thm p ng
vim. V vy, NF-B l mt ch hin nhin cho cc kiu iu tr khng vim
mi. Glucococticoit l cc cht c ch NF-B hu hiu nhng chng c nhiu tc
dng ph ti ni tit v chuyn ha khi c s dng vi liu php iu tr ton
367

thn. Cc tc dng ph ny s khng xy ra khi s dng cc cht c ch NF-B

c hiu hn. V vy, vic tm kim v nghin cu k lng cc hp cht thin
nhin c tc dng c ch c hiu NF-B v ang thu ht mnh m s quan
tm v u t nghin cu ca cc nh khoa hc trn ton th gii.
Kt qu nghin cu ca chng ti (bng IV.2.5.1) cho thy, 04 hp cht
SB1 - SB4 th hin hot tnh khng NF-B tt. y u l cc hp cht c cu
trc c o, c trng cho loi sao bin Archaster typicus (c th s dng chng
nh yu t ch th trong phng php nh loi ha hc). Cc hp cht ny c
phn lp vi hm lng cao trong loi sao bin Archaster typicus, iu ny t c
s cho vic tin hnh cc nghin cu ton din hn loi sao bin ny theo nh
hng hot tnh khng NF-B. Mt iu rt ng quan tm na l SB1 l mt hp
cht c cu trc mi v c phn lp vi hm lng cao nht so vi cc cht cn
li, iu ny gi m hng nghin cu, pht trin cc dc cht mi khng yu t
phin m NF-B v c s dng iu tr cc bnh vim, gp phn nng cao
gi tr s dng ca ngun nguyn liu sinh vt bin qu bu v sn c ca nc
ta.
III.2.5.2. Kt qu nh gi hot tnh gy c t bo ca cc hp cht
Tt c cc cht phn lp c t loi sao bin Archaster typicus c tin
hnh nh gi hot tnh gy c t bo. Kt qu th nghim (bng IV.2.5.2.) cho
thy c 4 hp cht SB1 - SB4 th hin hot tnh gy c t bo. Trong hp
cht SB1 th hin hot tnh gy c t bo i trn c 3 dng t bo th nghim,
SB2 khng 2 dng t bo KB v Hep-G2 vi gi tr IC50 ln lt l 3,37 v 2,69
g/ml, hai hp cht cn li SB3 v SB4 ch khng dng Hep-G2 vi gi tr IC50
ln lt l 4,52 v 3,72 g/ml.
Bng IV.2.5.2. Hot tnh gy c t bo ca cc cht
Hp cht
DMSO
Ellipticine
SB1
SB2
SB3
SB4
SB5
SB6
SB7

Gi tr CS% (Cell survival)

KB
1000,0
1,071,07
31,20,7
42,11,6
79,40,7
88,61,3
83,12,2
72,21,1
69,73,1

FL
1000,0
1,520,00
41,92,5
84,23,2
74,22,2
69,93,7
74,70,9
69,20,6
94,52,2

Hep-G2
100,00,0
1,340,63
24,50,6
39,72,1
44,61,7
37,53,3
85,24,2
59,72,8
76,32,5

IC50 (g/ml)
KB
0,21
4,45
3,37
(-)
(-)
(-)
(-)
(-)

FL
0,12
3,91
(-)
(-)
(-)
(-)
(-)
(-)

Hep-G2
0,19
2,35
2,69
4,52
3,72
(-)
(-)
(-)

Cc hp cht ny u c phn lp vi hm lng ln, c cu trc c

th (c bit hp cht SB1 l mt hp cht mi) v cn th hin c hot tnh
khng yu t phin m NF-B, iu ny t c s cho vic tin hnh cc nghin

368

cu ton din hn v cc hp cht ny nhm pht trin cc dc cht mi t

ngun dc liu bin qu bu v di do ca nc ta.
IV.2.6. Cc cng trnh cng b
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc
loi sao bin Archaster typicus, chng ti bin son v ng ti 02 cng trnh
trn cc tp ch chuyn ngnh quc gia, 01 cng trnh ti Hi ngh Khoa hc
Quc gia v 01 cng trnh ti Hi ngh Khoa hc Quc t l:
(1) Chau Van Minh, Nguyen Xuan Cuong, Nguyen Van Thanh, Hoang Thanh Huong, and
Phan Van Kiem, Highly oxygenated sterols from the starfish Archaster typicus. Tp
ch Ha hc, 2007, 45(3), 377-381.
(2) Phan Van Kiem, Nguyen Van Thanh, Nguyen Xuan Cuong, and Chau Van Minh,
Cholest-7-en-3-one, cholesterol and batilol from the starfish Archaster typicus. Tp ch
Ha hc, 2007, 45(4), 513-517.
(3) Chu Vn Minh, Nguyn Vn Thanh, Nguyn Xun Cng, Phan Vn Kim,

Nunziatina De Tommasi, Lm Ngc Thim. Mt galactocereboside mi t sao

bin Archaster typicus. Hi ngh Ha hc hu c Ton quc ln th 4, H Ni,
thng 10 nm 2007, trang 446-450.
(4) Chau Van Minh, Nguyen Van Thanh, Nguyen Xuan Cuong, Phan Van Kiem,
Nunziatina De Tommasi. A new galactocereboside from the starfish Archaster
typicus, 12t Asian Chemical Congress (12ACC), August 23-25, 2007, Malaysia,
p. 309.

Cc kt qu rn c s dng trong 01 lun n tin s Ha hc v

bo v thnh cng vi ti: Nghin cu thnh phn ha hc v kho st hot
tnh gy c t bo in vitro ca loi hi sm Holothuria scabra v sao bin
Archaster typicus sinh sng ti vng bin Vit Nam, TS. Nguyn Vn Thanh.

369

Tng hp cc cht phn lp c t mu sao bin

Archaster typicus
OH
21

4"

H
HO

2'

1'

6"

12

H O

5"

2"

3"

1"

OH
H

OH

NH
2

10

16
14

15

OH

OH

HO

(CH2)9CH(CH3)2

Gal

23

17

13

OH

11

19

(CH2)21CH3

OH
OH

OH

OH

SB2: 27-Norcholestan3,4,5,6,8,14,15,24R-octol

SB1, Archasterserebroside (cht mi)

28

OH
21

22

12
11

19

1
3

23

19

15
2

OH

OH

22

18

13
14

2
3

HO

1
10
4

OH
27

16
14

15

OH

OH

OH

SB4: Ergost-22-en3,4,5,6,8,14,15,22E,24R,25R,
26-nonol

20

24

21

26

27

17

22

18

25

23

19

16
15

12

11

13
14

8
6

CH 2 OH

OH

21

11

OH

SB3: 27-Norcholestan3,4,5,6,7,8,14,15,24R-nonol

19

10

23

17

13

OH

HO

OH

12

OH

OH
OH

11

26
25

24

20

12

16
14

22

18

HO

26

25

10

21

17

13

OH

24

20

18

26

25

24

20

18

OH
OH OH

22

SB5: Cholesterol

1
10
4

20

24
23

17
16
15

8
7

SB6: Cholest- 7- en- 3- on

1

OH
HO

1'

SB7: Batilol

370

18'

CH2(CH2)16CH3

26
25
27

Chng V. Nghin cu ho hc theo nh hng

hot tnh sinh hc ca mt s loi Hi min
V.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca
loi hi min xestospongia testudinaria (petrosidae)
V.1.1. M u
Mu hi min Xestospongia
testudinaria c thu thp ti Nam
Yt, Qun o Trng Sa, tnh
Khnh Ha vo thng 03 nm 2007
v c TS. Cng Thung v cc
cng s, Vin Ti nguyn v Mi
trng Bin, Vin KH&CN Vit
Nam gim nh tn khoa hc.
- M t: hnh dng, mu sc v kt
cu b mt thay i rt nhiu tu
thuc vo mi trng sng, thng
chng c dng cc ln vi b mt
nhiu cc g v cc rnh, cu trc
cng v d v vn.

Hi min Xestospongia testudinaria

Hi min (sponge) l ng vt a bo trung gian gm cc c th a bo

nhng khng t chc thnh m hay c quan. iu ny ni ln rng mc d l c
th a bo nhng nhng t bo ny kt hp li thnh cu trc c th c bit, cc
c th ny thiu h thng thn kinh v tun hon. Do vy, n c tnh cht ca mt
tp on ng vt nguyn sinh hn mc d c nhiu loi t bo hn. ng vt a
bo trung gian c coi l ngun gc chung ca cc ng vt a bo trong qu
trnh tin ha cn cc ng vt nguyn sinh l mt nhnh c lp khng lin
quan g n s tin ha ca ng vt a bo. Hi min mc d l mt c th c t
chc c th a bo, d cc t bo c th kt hp li vi nhau thnh hai lp t bo,
nhng mi t bo vn c th sng mt cch c lp khi b tch ri.
Xestospongia l mt chi hi min sinh sng kh ph bin ti cc vng bin
Vit Nam. Hin nay nc ta hon ton cha c cng trnh khoa hc no c
cng b v thnh phn ha hc v hot tnh sinh hc ca cc loi thuc chi ny.
Trn th gii hin cng c rt t cc cng trnh cng b v chi ny v mi c 12
hp cht c phn lp v xc nh cu trc. Hai thnh phn ha hc chnh ca
loi ny l cc axit bo khng no b brom ha v cc hp cht steroit. Nm 1985,
nhm nghin cu ca tc gi Ronald J. Q. cng b s phn lp v xc nh cu
trc ca 1 axit bo khng no b brom ha l l 16-Bromo-7E,15E-hexadecadiene5,13-diynoic acid [246]. Nm 1991, nhm nghin cu ny li cng b thm 3 hp
cht dng ny [247]. Gn y nht (1999), 2 hp cht xestosterol este ca axit 18bromooctadeca-(9E,17E)-diene-7,15-diynoic v xestosterol este ca axit 18bromooctadeca-(9E,17E)-diene-5,7,15-triynoic c phn lp t loi ny [248].

371

Di y, chng ti gii thiu mt s hp cht quan trng c phn

lp t loi Xestospongia testudinaria.
COOH

Br

16-Bromo-7E,15E-hexadecadiene-5,13-diynoic acid
COOH

Br

16-Bromo-7E,11E,15Z-hexadecatriene-5,13-diynoic acid
COOCH3

Br

Metyl ester ca axit 18-bromo-9E,17E-octadecadiene-5,7,15-triynoic

COOH
Br

Br

18,18-Dibromo-9E,17-octadecadiene-5,7-diynoic acid

Br

O
O

Xestosterol este ca axit 18-bromooctadeca-(9E,17E)-diene-7,15-diynoic

Br
O
O

Xestosterol este ca axit 18-bromooctadeca-(9E,17E)-diene-5,7,15-triynoic

372

V.1.2. Phn lp cc hp cht

Hi min X. testudinaria (15 kg ti) c ra sch, xay nh v chit bng
metanol (3 ln) bng thit b chit siu m. Dch metanol ca cc ln chit c
lc, gom li v ct loi dung mi nhit khng qu 60oC di p sut gim
thu c 120 g dch cn dch metanol. Cn dch ny c ha vo nc v chit
phn b ln lt vi cc dung mi clorofooc v butanol theo t l 1/1 (3 ln, mi
ln 6 lit) thu c cc cn dch tng ng l clorofooc 60 g (k hiu: C), butanol
40 g (k hiu B) v nc 20 g (k hiu N) (S V.1.2.a).
Cn chit clorofrom: C (60 g) c tin hnh chy ct th trn sc k ct
pha thng gradient nng metanol trong cloroform thu c 5 phn on nh
hn l C1-5. Cc phn on C1, C2 v C3 c tip tc phn tch trn sc k ct
pha thng vi h dung mi ra gii thch hp thu c cc hp cht XT1 n
XT6 v XT9 - XT10 nh c ch ra trn S V.1.2.c. Bng cc phng php
tng t (S V.1.2.b), cc hp cht XT7, v XT8 c phn lp t cn chit
BuOH.
Hi min Xestospongia
testudinaria ti (15 kg)

- Chit MeOH, ct loi dung mi di p sut

gim

B sung nc
B sung clorofooc

GM - Cn chit MeOH (120g)

clorofooc: nc 1/1

Lp nc
B sung BuOH

Cn clorofooc
C (60 g)

BuOH: nc 1/1

Lp nc

Cn nc
N (20 g)

Cn BuOH
B (40 g)

S V.1.2.a: S chit phn on mu hi min

373

B - (40 g)

Silica gel CC
Cloroform/MeOH/H2O: 5/1/0 - 2/1/0.2

B1 (1.0 g)

B2 (20 g)

B3 (3 g)

Silica gel CC
CHCl3/MeOH/H2O
5/1/0.1

B4 (6 g)

B5 (10 g)

Silica gel CC
CHCl3/MeOH/H2O
4/1/0.1

XT8
(33 mg)

XT7
(40mg)

S V.1.2.b: S phn lp cc hp cht t cn chit butanon

C (60 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

C1 (30g)

C2 (5 g)

C3 (7 g)

C4 (3 g)

C5 (15 g)

Silica gel CC
n-hexan/ axeton 4/1

XT9
(12mg)

XT10
(8mg)

Silica gel CC
n-hexan/clorofooc/axeton 6/1/0,3

XT4
(20 mg)

XT5
(80 mg)

XT6
(35 mg)

XT2
(30 mg)

XT3
(1200 mg)

Silica gel CC
metanol/nc 5/1

XT1
(20 mg)

S V.1.2.c: S phn lp cc hp cht t cn chit cloroform

374

V.1.3. Hng s vt l v d kin ph ca cc hp cht

V.1.3.1. Hp cht XT1: axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17triynoic
Kt tinh mu trng, ph khi lng ESI-MS: m/z 394.8/396.8 [M+Na]+ vi t l
1/0,98 v cng tn hiu (C20H21BrO2, M = 372).
1
H-NMR (500 MHz, CDCl3) (ppm): 2,36 (2H, t, J = 7,5 Hz, H-2), 1,65 (2H, m,
H-3), 1,47 (2H, m, H-4), 1,57 (2H, m, H-5), 2,33 (2H, t, J = 7,0 Hz, H-6), 5,52 (1H, d, J
= 15,5 Hz, H-11), 6,23 (1H, m, H-12), 2,23 (2H, m, H-13), 2,23 (2H, m, H-14), 6,13
(1H, m, H-15), 5,57 (1H, dd, J = 15,5, 1,5 Hz, H-16), 6,29 (1H, dd, J = 14,0, 2,5 Hz, H19) v 6,65 (1H, d, J = 14,0 Hz, H-20).
13
C-NMR (125 MHz, CDCl3) (ppm): 179,29 (C-1), 33,75 (C-2), 24,12 (C-3),
28,19 (C-4), 27,87 (C-5), 19,38 (C-6), 83,52 (C-7), 73,68 (C-8), 65,40 (C-9), 73,43 (C10), 109,84 (C-11), 146,02 (C-12), 32,15 (C-13), 32,15 (C-14), 143,83 (C-15), 110,32
(C-16), 90,22 (C-17), 85,04 (C-18), 117,64 (C-19) v 117,89 (C-20).
V.1.3.2. Hp cht XT2: axit 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19-triynoic
Kt tinh mu trng, ph khi lng ESI-MS: m/z 424.9/426.9 [M+Na]+ vi t l
1/0,98 v cng tn hiu (C22H27O2Br, M = 402).
1
H-NMR (500 MHz, CDCl3) (ppm): 2,35 (2H, t, J = 7,5 Hz, H-2), 1,63 (2H, m,
H-3), 1,41 (2H, m, H-4), 1,37 (2H, m, H-5), 1,39 (2H, m, H-6), 1,51 (2H, m, H-7), 2,25
(4H, t, J = 7,0 Hz, H-8 v H-13), 1,51 (4H, m, H-14 v H-15), 2,14 (2H, m, H-16), 6,17
(1H, dt, J = 15,5, 7,0 Hz, H-17), 5,55 (1H, dd, J = 15,5, 1,5 Hz, H-18), 6,30 (1H, dd, J
= 14,0, 2,0 Hz, H-21) v 6,63 (1H, d, J = 14,0 Hz, H-22).
13
C-NMR (125 MHz, CDCl3) (ppm): 179,39 (C-1), 33,89 (C-2), 24,61 (C-3),
28,87 (C-4), 28,70 (C-5), 28,60 (C-6), 28,12 (C-7), 19,01 (C-8), 77,62 (C-9), 65,64 (C10), 65,27 (C-11), 77,92 (C-12), 19,17 (C-13), 27,66 (C-14), 27,68 (C-15), 32,64 (C-16),
145,29 (C-17), 109,60 (C-18), 90,52 (C-19), 84,65 (C-20), 117,73 (C-21) v 117,65 (C22).
V.1.3.3. Hp cht XT3: cholesterol
Cht kt tinh mu trng, im chy 148-149 oC, C27H46O, M=386.
H-NMR (500 MHz, CDCl3) (ppm): 3,52 (1H, m, H-3), 5,35 (1H, br d, J = 5,0
Hz, H-6), 1,00 (3H, s, H-19), 0,91 (3H, s, H-21), 0,86 (6H, d, J = 6.5 Hz), H-26, 27) and
0.68 (3H, s, H-18).
1

C-NMR (125 MHz, CDCl3) (ppm): 37.2 (t, C-1), 31.6 (t, C-2), 71.8 (d, C-3),
42,2 (t, C-4), 140,7 (s, C-5), 121,7 (d, C-6), 31,8 (t, C-7), 31,8 (d, C-8), 50,1 (d, C-9),
36,5 (s, C-10), 21,1 (t, C-11), 39,8 (t, C-12), 42,3 (s, C-13), 56,7 (d, C-14), 24,3 (t, C15), 28,3 (t, C-16), 56,1 (d, C-17), 11,9 (q, C-18), 19,4 (q, C-19), 35,7 (d, C-20), 18,7 (q,
C-21), 36,1 (t, C-22), 23,8 (t, C-23), 39,5 (t, C-24), 28,0 (d, C-25), 22,5 (q, C-26), 22,8
(q, C-27).
13

V.1.3.4. Hp cht XT4: 5,8-epidioxycholest-6-en-3-ol

Tinh th mu trng, im chy 102 - 105oC, ph khi lng ESI-MS m/z: (positive)
417.0 [M+H]+, 439.0 [M +Na]+, C27H44O3 (M = 416).
1
H-NMR (CDCl3, 500 MHz) (ppm): 1,94 (1H, m, Ha-1), 1,99 (1H, m, Hb-1), 1,54
(1H, m, Ha-2), 1,83 (1H, m, Hb-2), 3,96 (1H, m, H-3), 1,95 (1H, m, Ha-4), 2,11 (1H, m,
375

Hb-4), 6,24 (1H, d, J = 8,5 Hz, H-6), 6,51 (1H, d, J = 8,5 Hz, H-7), 1,52 (1H, m, H-9),
1,00 (1H, m, Ha-11), 1,59 (1H, m, Hb-11), 1,22 (2H, m, H-12), 1,58 (1H, m, H-14),
1,21(1H, m, Ha-15), 1,52 (1H, m, Hb-15), 1,45 (2H, m, H-16), 1,20 (1H, m, H-17), 0,80
(3H, s, H-18), 0,88 (3H, s, H-19), 1,60 (1H, m, H-20), 0,90 (3H, d, J = 7,0 Hz, H-21),
1,70 (2H, m, H-22), 1,32 (2H, m, H-23), 1,24 (2H, m, H-24), 1,42 (1H, m, H-25), 0,86
(3H, d, J = 7,0 Hz, H-26), v 0,87 (3H, d, J = 7,0 Hz, H-27).
13
C-NMR (CDCl3, 125 MHz) (ppm): 34,71 (C-1), 30,11 (C-2), 66,44 (C-3),
36,94 (C-4), 82,16 (C-5), 135,40 (C-6), 130,77 (C-7), 79,46 (C-8), 51,09 (C-9), 36,95
(C-10), 20,62 (C-11), 39,44 (C-12), 44,74 (C-13), 51,59 (C-14), 23,42 (C-15), 28,23 (C16), 56,44 (C-17), 12,62 (C-18), 18,58 (C-19), 35,22 (C-20), 18,16 (C-21), 35,95 (C-22),
23,80 (C-23), 39,44 (C-24), 27,98 (C-25), 22,53 (C-26), v 22,79 (C-27).
V.1.3.5. Hp cht XT5: cholest-7-en-3-one.
Tinh th mu trng, im chy 147-148oC, ph khi lng ESI-MS: m/z 385,2
+
[M+H] (C27H44O, M = 384).
H-NMR (500 MHz, CDCl3) (ppm): 1,47 (1H, m, Ha-1), 2,15 (1H, ddd, J =
4,5, 9,2, 13,6 Hz, Hb-1), 2,28 (1H, br d, J = 14,5 Hz, Ha-2), 2,42 (1H, dt, J = 6,0, 14,5
Hz, Hb-2), 5,18 (1H, br s, H-7), 0,57 (3H, s, H-18), 1,03 (3H, s, H-19), 1,39 (1H, m, H20), 0,93 (3H, d, J = 6,5 Hz, H-21), 0,86 (3H, d, J = 7,0 Hz, H-26), 0,87 (3H, d, J = 7,0
Hz, H-27).
1

C-NMR (125 MHz, CDCl3) (ppm): 38,8 (C-1), 38,1 (C-2), 211,8 (C-3), 44,2
(C-4), 42,9 (C-5), 30,1 (C-6), 116,9 (C-7), 139,5 (C-8), 48,9 (C-9), 34,4 (C-10), 21,7 (C11), 39,5 (C-12), 43,4 (C-13), 54,9 (C-14), 22,9 (C-15), 27,9 (C-16), 56,2 (C-17), 11,9
(C-18), 12,4 (C-19), 36,2 (C-20), 18,9 (C-21), 36,1 (C-22), 23,9 (C-23), 39,5 (C-24),
28,1 (C-25), 22,5 (C-26) v 22,8 (C-27).
V.1.3.6. Hp cht XT6: saringosterol
13

Tinh th hnh kim mu trng; im chy 160-161oC; ph khi lng ESI-MS:

m/z 411,1 [M-H2O+H]+, 393,0 [M-2H2O+H]+ (C29H49O2, M = 428).
1
H-NMR (500 MHz, CDCl3) (ppm): 1,07 (1H, Ha-1), 1,85 (1H, Hb-1), 1,50
(1H, Ha-2), 1,82 (1H, Hb-2), 3,52 (1H, m, H-3), 2,27 (2H, m, H-4), 5,35 (1H, t, J = 3,0
Hz, H-6), 1,52 (1H, Ha-7), 1,97 (1H, Hb-7), 1,45 (1H, m, H-8), 0,95 (1H, m, H-9), 1,49
(2H, m, H-11), 1,18 (1H, Ha-12), 2,20 (1H, Hb-12), 0,99 (1H, m, H-14), 1,05 (1H, Ha15), 1,58 (1H, Hb-15), 1,26 (1H, Ha-16), 1,82 (1H, Hb-16), 1,15 (1H, m, H-17), 0,68
(3H, s, H-18), 1,00 (3H, s, H-19), 1,39 (1H, m, H-20), 0,92 (3H, d, J = 7,0 Hz, H-21),
1,03 (1H, Ha-22), 1,41 (1H, Hb-22), 1,58 (1H, Ha-23), 1,62 (1H, Hb-23), 1,73 (1H, m, H25), 0,90 (3H, d, J = 7,0 Hz, H-26), 0,87 (3H, d, J = 7,0 Hz, H-27), 5,80 5,18 (1H, dd J
= 1,5 v 17,5 Hz, Hb-29). (1H, dd, J = 10,5 v 17,5 Hz, H-28) v 5,13 (1H, dd J = 1,5
v 10,5 Hz, Ha-29).
13
C-NMR (125 MHz, CDCl3) (ppm): 37,27 (C-1), 31,67 (C-2), 71,81 (C-3),
42,31 (C-4), 140,77 (C-5), 121,70 (C-6), 31,91 (C-7), 31,91 (C-8), 50,13 (C-9), 36,52
(C-10), 21,08 (C-11), 39,76 (C-12), 42,34/42,31 (C-13), 56,75 (C-14), 24,28 (C-15),
28,22/28,17 (C-16), 55,83 (C-17), 11,86 (C-18), 19,40 (C-19), 36,10/36,14 (C-20),
18,79 (C-21), 29,10/29,07 (C-22), 34,57/34,79 (C-23), 77,71 (C-24), 35,93 (C-25),
16,47 (C-26), 16,54 (C-27), 142,57/142,4 (C-28) v 112,95/112,86 (C-29).
376

V.1.3.7. Hp cht XT7: thymidine

Tinh th mu trng, im chy 186-187oC, ph khi lng ESI-MS: m/z 264,7
[M+Na]+, 240.8 [M-H]- (C10H14N2O5, M = 242).
1
H-NMR (500 MHz, CD3OD) (ppm): 7,82 (1H, d, J = 1,0 Hz, H-6), 1,90 (3H,
d, J = 1,0 Hz, H-7), 4,41 (1H, m, H-1), 2,24 (1H, m, H-2), 3,92 (1H, dd, J = 3,5, 6,5
Hz, H-3), 6,29 (1H, m, H-4), 3,75 (2H, m, H-5).
13
C-NMR (125 MHz, CD3OD) (ppm): 152,4 (C-2), 166,4 (C-4), 111,5 (C-5),
138,2 (C-6), 12,4 (C-7), 86,3 (C-1), 41,2 (C-2), 72,2 (C-3), 88,8 (C-4) v 62,8 (C-5).
V.1.3.8. Hp cht XT8: thymine
Tinh th mu trng, im chy 326oC, ph khi lng ESI-MS: m/z 126.8
+
[M+H] , 148.7 [M+Na]+, 124.7 [M-H]- (C5H6N2O2, M = 126).
1
H-NMR (500 MHz, DMSO-d6) (ppm): 10,56 (1H, br s, H-1), 10,97 (1H, br s,
H-3), 7,24 (1H, d, J = 1,0 Hz, H-6) v 1,72 (3H, d, J = 1,0 Hz, H-7).
V.1.3.9. Hp cht XT9: chimyl alcohol
Tinh th mu trng, im chy 64-65oC; ph khi lng ESI-MS m/z: 317,1
[M+H] , (C19H40O3, M=316).
+

H-NMR (500 MHz, CDCl3) (ppm): 3,64 (1H, dd, J = 3,0 v 11,0 Hz, Ha-1),
3,71 (1H, dd, J = 5,0 v 11,0 Hz, Hb-1), 3,86 (1H, m, H-2), 3,52 (2H, m, H-3), 3,45 (2H,
m, H-1), 1,58 (2H, m, H-2), 1,26 (26H, br s, H-3 n H-15), 0,88 (3H, t, J = 7,0 Hz,
H-16)
1

C-NMR (125 MHz, CDCl3) (ppm): 72,5 (C-1), 70,4 (d, C-2), 64,3 (C-3), 71,9
(C-1), 29,3 - 29,7 (C-2), 26,1 (C-3), 29,3 - 29,7 (C-4 n C-13), 31,9 (C-14), 22,7
(C-15), 14,2 (C-16).
13

V.1.3.10. Hp cht XT10: batilol

Tinh th mu trng; im chy 70-71oC; ph khi lng ESI-MS m/z: 345,1
[M+H]+, (C21H44O3, M = 344).
1
H-NMR (500 MHz, CDCl3) (ppm): 3,62 (1H, dd, J = 3,0 v 11,0 Hz, Ha-1),
3,71 (1H, dd, J = 5,0 v 11,0 Hz, Hb-1), 3,86 (1H, m, H-2), 3,52 (2H, m, H-3), 3,45 (2H,
m, H-1), 1,57 (2H, m, H-2), 1,26 (30H, br s, H-3 n H-17) v 0,88 (3H, t, J = 7,0
Hz, H-18).

377

V.1.4. Xc nh cu trc ha hc ca cc hp cht

V.1.4.1. Hp cht XT1: axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene7,9,17-triynoic
Hp cht XT1 phn lp c di dng tinh th hnh kim mu trng. Cng
thc phn t ca n c xc nh l C20H21BrO2 (M = 372) bng cc kt qu ph
khi lng (ESI-MS ti m/z 394,8/396,8 [M+Na]+ vi t l 1/0,98 v cng tn
hiu) v cc ph cng hng t ht nhn (1H-, 13C-NMR v cc ph DEPT). Hp
cht ny rt km bn nn i hi phi c o ph khi lng v ph cng hng
t ht nhn ngay sau khi c lm sch.
Trn ph 1H-NMR xut hin cc tn hiu c trng ca ba ni i ngoi
vng u b th hai v tr ti 5,52 (1H, d, J = 15,5 Hz, H-11), 5.57 (1H, dd, J =
15.5, 1.5 Hz, H-16), 6.13 (1H, m, H-15), 6.23 (1H, m, H-12), 6.29 (1H, dd, J =
14.0, 2.5 Hz, H-19) v 6.65 (1H, d, J = 14.0 Hz, H-20). S xut hin ca by
nhm metilen c c trng bi cc tn hiu cng hng ti 1,47 (2H, m, H4), 1,57 (2H, m, H-5), 1,65 (2H, m, H-3), 2,23 (4H, m, H-13 v H-14), 2,33 (2H,
t, J = 7,0 Hz, H-6) v 2,36 (2H, t, J = 7,5 Hz, H-2).

Hnh V.1.4.1.a. Ph 1H-NMR ca XT1

6
7
9
17
15

11

Br
19

Hnh V.1.4.1.b. Cu trc ha hc ca XT1

378

COOH
1

Ph 13C-NMR ca XT1 xut hin cc tn hiu ca 20 cacbon trong c 7

cacbon bc bn, 7 cacbon metilen v 6 cacbon metin. Ba lin kt i ngoi vng
mt ln na c khng nh bi cc tn hiu cacbon metin ti 109.84 (C-11),
110,32 (C-16), 117,64 (C-19), 117,89 (C-20), 143,83 (C-15) v 146,02 (C-12).
Ngoi ra, s xut hin ca 3 lin kt ba c c trng bi cc tn hiu u l
cacbon bc bn cng hng ti 65,40 (C-9), 73,43 (C-10), 73,68 (C-8), 83,52
(C-7), 85,04 (C-18) v 90,22 (C-17). Mt nhm cacbonyl c xc nh bi tn
hiu cng hng ti 179,29 (C-1). Cc nhm metilen nm v tr v so vi
nhm cacbonyl c khng nh bi cc tn hiu cng hng tng ng ti
33,75 (C-2) v 24,12 (C-3). Mt nhm metilen nm cnh ni ba v 2 nhm
metilen nm cnh ni i c khng nh bi cc tn hiu tng ng ti 19,38
(C-6), 32,15 (C-13 v C-14).

Hnh V.1.4.1.c. Ph 13C-NMR ca XT1

Hnh V.1.4.1.d. Ph 13C-NMR v cc ph DEPT ca XT1

379

T cc phn tch nu trn, c th s b xc nh XT1 l mt hp cht axit

mch di b brm ha c cha 3 ni i v 3 ni ba. Cu trc hnh hc ca cc
ni i c xc nh u dng trans bi hng s tng tc ln (J = 14,0 v
15,5 Hz) ca cc proton.

Hnh V.1.4.1.e. Ph HSQC ca XT1

Bng V.1.4.1. Kt qu ph NMR ca XT1
C

C [249]

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20

179,4
36,0
25,7
19,7
83,7
74,4
66,5
74,0
111,2
147,4
33,1
33,1
145,3
110,0
91,1
85,6
118,7
118,9

179,29
33,75
24,12
28,19
27,87
19,38
83,52
73,68
65,40
73,43
109,84
146,02
32,15
32,15
143,83
110,32
90,22
85,04
117,64
117,89

Ha, c
dng pic (J = Hz)
2,36 t (7,5)
1,65 m
1,47 m
1,57 m
2,33 t (7,0)
5,52 d (15,5)
6,23 m
2,23 m
2,23 m
6,13 m
5,57 dd (15,5, 1,5)
6,29 dd (14,0, 2,5)
6,65 d (14,0)

o trong CDCl3, b125 MHz, c500 MHz

380

HMBC
(H C)
1, 3, 4
1, 2, 3
2, 5, 6
3, 4, 6, 7
5, 7, 8

9, 10, 12, 13
10, 13
11, 12, 14
13, 15, 16
14, 17
14, 15, 18
17, 20
18, 19

Hnh V.1.4.1.f. Ph 1H-1H COSY ca XT1

V tr th ca nguyn t brm ti C-20 v s xp xp cc lin kt i v
lin kt ba ca XT1 nh c ch ra Hnh V.1.4.1.b c khng nh bng cch
so snh s liu ph NMR ca n vi cc s liu ph c cng b ca hp
cht c cu trc tng t [249]. iu ny mt ln na c khng nh bi cc
tng tc trn ph cng hng t ht nhn hai chiu HMBC v 1H-1H COSY. Cc
phn tch chi tit c ch ra trong Bng V.1.4.1 v Hnh V.1.4.1.h.

Hnh V.1.4.1.g. Ph HMBC ca XT1

381

COOH

9
17
15

Br

11

19

COSY
HMBC

Hnh V.1.4.1.h. Cc tng tc HMBC (HC) v COSY chnh ca XT1

Hnh V.1.4.1.i. Ph khi lng ESI-MS ca XT1

T tt c cc phn tch nu trn, cu trc ha hc ca XT1 c xc nh
l axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic. y l
mt hp cht c cu trc mi, ln u tin c phn lp t thin nhin. Theo ti
liu cng b [249], cc hp cht axit mch di a ni i v ni ba thng c
hot tnh khng sinh v gy c t bo. c bit, cng theo ti liu ny, mt hp
cht c cu trc tng t nh XT1 ch ngn hn 2 nhm metilen phn u
mch th hin hot tnh c ch enzym HIV protease, mt enzym thit yu cho s
ti bn ca HIV. iu ny gi rng XT1 cng c th c hot tnh tng t.
V.1.4.2. Hp cht XT2: axit 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19triynoic
Hp cht XT2 cng c phn lp di dng tinh th hnh kim mu trng.
Cng thc phn t ca n c xc nh l C22H27O2Br (M = 402) bng cc d

382

kin ph khi lng (ti m/z 424,9/426,9 [M+Na]+ vi t l 1/0,98 v cng

tn hiu) v ph cng hng t ht nhn (1H-, 13C-NMR v cc ph DEPT). Hp
cht ny cng rt km bn nn i hi phi c o ph khi lng v ph cng
hng t ht nhn ngay sau khi c lm sch.

Hnh V.1.4.2.a. Ph 1H-NMR ca XT2

8
9

COOH

11

19
17

22

Br

13

21

Hnh V.1.4.2.b. Cu trc ha hc ca XT2

Ph 1H-NMR ca XT2 c dng tng t nh ca XT1. iu ny cho thy
hai hp cht c cu trc tng t nhau. S khc bit d nhn thy nht trn ph
1
H-NMR ca XT2 so vi XT1 l s mt i tn hiu ca mt ni i v s xut
hin thm tn hiu ca 2 nhm metilen. Hai ni i c khng nh bi cc tn
hiu cng hng ti 5,55 (1H, dd, J = 15,5, 1,5 Hz, H-18), 6,17 (1H, dt, J =
15,5, 7,0 Hz, H-17), 6,30 (1H, dd, J = 14,0, 2,0 Hz, H-21) v 6,63 (1H, d, J =
14,0 Hz, H-22). Cc tn hiu metilen xut hin ti vng trng cao: 2,35 (2H, t,
J= 7,5 Hz, H-2), 1,63 (2H, m, H-3), 1,41 (2H, m, H-4), 1,37 (2H, m, H-5), 1,39
(2H, m, H-6), 1,51 (2H, m, H-7), 2,25 (4H, t, J = 7,0 Hz, H-8 v H-13), 1,51 (4H,
m, H-14 v H-15) v 2,14 (2H, m, H-16).

383

Hnh V.1.4.2.c. Ph 13C-NMR ca XT2

Hnh V.1.4.2.d. Ph 13C-NMR v cc ph DEPT ca XT2

384

Ph 13C-NMR ca XT2 c dng tng t nh ca XT1 vi s xut hin

cc tn hiu ca 22 cacbon trong c 7 cacbon bc bn, 4 cacbon metin v 11
cacbon metilen. S xut hin ca nhm cacbonyl c khng nh bi tn hiu
cng hng ti 179,39 (C-1). Cc tn hiu cng hng ti 109,60 (C-18),
117,65 (C-22), 117,73 (C-21) v 145,29 (C-17) khng nh s tn ti ca 2 lin
kt i. S c mt ca 3 lin kt ba c khng nh bi cc tn hiu cng hng
ti 65,27 (C-11), 65,64 (C-10), 77,62 (C-9), 77,92 (C-12), 84,65 (C-20) v
90,52 (C-19). Cc tn hiu cng hng ti 33,89 (C-2) v 24,61 (C-3) c trng
tng ng cho cc nhm metilen nm v tr v so vi nhm cacbonyl. Hai
nhm metilen nm cnh lin kt ba v mt nhm metilen nm cnh lin kt i
cng c khng nh bi cc tn hiu cng hng tng ng ti 19,01 (C-8),
19,17 (C-13) v 32,64 (C-16).

Hnh V.1.4.2.e. Ph HSQC ca XT2

Cc tn hiu proton c gn vi cacbon tng ng thng qua cc tn hiu
tng tc trn ph cng hng t ht nhn hai chiu tng tc trc tip (HSQC).
Kt qu chi tit c ch ra trong Bng V.1.4.2.
Bng V.1.4.2. Kt qu ph NMR ca XT2
C

Ca, b

1
2
3
4
5

179,39
33,89
24,61
28,87
28,70

Ha, c
dng pic (J = Hz)
2,35 t (7,5)
1,63 m
1,41 m
1,37 m
385

HMBC
(H C)
1, 3, 4
1, 2, 4, 5

6
28,60
1,39 m
7
28,12
1,51 m
8
19,01
2,25 t (7,0)
9
77,62
10
65,64
11
65,27
12
77,92
13
19,17
2,25 t (7,0)
14
27,66
1,51 m
15
27,68
1,51 m
16
32,64
2,14 m
17
145,29
6,17 dt (15,5, 7,0)
18
109,60
5,55 dd (15,5, 1,5)
19
90,52
20
84,65
21
117,73
6,30 dd (14,0, 2,0)
22
117,65
6,63 d (14,0)
a
b
o trong CDCl3, 125 MHz, c500 MHz

4, 5, 8
5, 6, 8, 9
6, 7, 9, 10

11, 12, 14, 15

12, 13, 15
13, 14
14, 15, 17, 18
15, 16, 19
16, 17, 20

19, 22
20, 21

Hnh V.1.4.2.f. Ph HMBC ca XT2

386

8
9

COOH

11

19
17
22

Br

13

21

Hnh V.1.4.2.g. Cc tng tc HMBC (HC) chnh ca XT2

V tr th ca nguyn t brm ti C-22 v s b tr cc lin kt i v lin
kt ba (Hnh V.1.4.2.b.) c xc nh bng cc tng tc trn ph cng hng t
ht nhn hai chiu tng tc xa (HMBC) v s so snh s liu ph 1H- v 13CNMR ca cc phn cu trc tng ng vi cc s liu c cng b ca cc
hp cht c cu trc tng t [249, 250]. Kt qu chi tit c ch ra trn Hnh
V.1.4.2.g v Bng V.1.4.2.

Hnh V.1.4.2.f. Ph khi lng ESI-MS ca XT2

T tt c cc phn tch nu trn, cu trc ha hc ca XT2 c xc nh
l axit 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19-triynoic. y l mt
hp cht c cu trc mi, ln u tin c phn lp t thin nhin.
V.1.4.3. Hp cht XT3: cholesterol
Hp cht XT3 nhn c di dng tinh th hnh kim c mu trng. Th
nghim trn sc k lp mng (TLC) vi thuc th c hiu (phun dung dch
H2SO4 10% ri h nng t t cho mu tm hng, sau chuyn dn sang tm
xanh) cho thy y l mt steroit. Hp cht ny thu c vi hm lng rt cao

387

cng vi gi tr rf = 0,35 trn TLC vi h dung mi l Hexan/Axeton 7/1 v gi

tr nhit nng chy (mp 148,5oC) ca n gi ti hp cht cholesterol, mt
sterol ph bin nht trong gii ng vt. Cng vi s ph hp hon ton v s liu
ph 1H- v 13C-NMR (xem phn 2.5.5) ca XT3 so vi ti liu cng b [220]
cho php khng nh hp cht ny chnh l cholesterol.
21

26
20

18

19

11

13
14

HO

17

25

27

15

10
3

22

7
6

Hnh V.1.4.3. Cu trc ha hc ca XT3

V.1.4.4. Hp cht XT4: 5,8-epidioxycholest-6-en-3-ol
Hp cht XT4 nhn c di dng cht tinh th hnh kim mu trng. Kt
qu ph khi lng ESI-MS (ti m/z: 417.0 [M+H]+ v 439.0 [M+Na]+) v ph
cng hng t ht nhn cho php xc nh cng thc phn t ca n l C27H44O3
(M = 416). Ph 1H-NMR ca XT4 c trng cho mt hp cht steroit, trong
xut hin cc tn hiu ca 5 nhm metyl ti 0,86 (3H, s), 0,90 (3H, d, J = 6,5
Hz), 0,90 (3H, d, J = 6,5 Hz), 0,92 (3H, s), 0,96 (3H, d, J = 6,5 Hz); cc proton
olefin ti 6,30 (1H, d, J = 8,5 Hz) v 6,55 (1H, d, J = 8,5 Hz); mt proton gn
vi cacbon c ni vi nguyn t xy ti 3,80 (1H, m).

Hnh V.1.4.4.a. Ph 1H-NMR ca XT4

388

Ph 13C-NMR xut hin cc tn hiu ca mt steroit 27 cacbon gm c 4

cacbon bc bn, 8 metin, 10 metilen v 5 metyl. Mt ni i b th 2 ln c
xc nh ti 130,77 (d)/135,40 (d); 2 cacbon bc 4 ni vi nguyn t oxy ti
79,46 (s) v 82,16 (s), 1 cacbon metin gn vi nguyn t xy ti 66,44 (d).
21
20

26

25

18

13

19
1

9
8

15

10
3

HO

27

17

11

Hnh V.1.4.4.b. Cu trc ha hc ca XT4

Bng V.1.4.4. Cc s liu ph NMR ca XT4
C

C a, b

Ha, c mult, (J = Hz)

34,71

1,94/1,99

30,11

1,54/1,83 (m)

66,44

3,96 (m)

36,94

1,95/2,11

82,16

135,40

6,24 (d, 8,5)

130,77

6,51 (d, 8,5)

79,46

51,09

1,52

10

36,95

11

20,62

1,00 (m)/1,59

12

39,44

1,22

13

44,74

14

51,59

1,58

15

23,42

1,21/1,52

16

28,23

1,45

17

56,44

1,20

18

12,62

0,80 (s)

19

18,58

0,88 (s)

389

20

35,22

1,60

21

18,16

0,90 (d, 7,0)

22

35,95

1,70 (m)

23

23,80

1,32 (m)

24

39,44

1,24

25

27,98

1,42

26

22,53

0,86 (d, 7,0)

22,79

0,87 (d, 7,0)

27
a

o trong CDCl3, 125MHz, 500MHz

Hnh V.1.4.4.c. Ph 13C-NMR ca XT4

T cc phn tch nu trn c th s b xc nh c XT4 c b khung
cacbon tng t nh ca XT3 tuy nhin c s chuyn dch v tr ca lin kt
i v s xut hin 2 cacbon bc bn gn vi nguyn t xy. chuyn dch ha
hc tng i cao ti cc v tr cacbon bc bn lin kt vi nguyn t xy gi
ti s c mt ca mt cu peroxit.

390

Hnh V.1.4.4.d. Ph 13C-NMR v cc ph DEPT ca XT4

Cui cng, cu trc ha hc ca XT4 c xc nh l 5,8epidioxycholest-6-en-3-ol bi s trng khp hon ton cc gi tr tng ng v
s liu ph 13C-NMR v cc c tnh vt l, ha hc ca XT4 vi hp cht ny
c cng b [251]. Hp cht ny cng c chng ti phn lp t loi cu
gai Diadema setosum v cc kt qu nghin cu v hot tnh sinh hc in vitro cho
thy hp cht ny th hin hot tnh gy c t bo cao i vi nhiu dng t bo
ung th nh dng KB (t bo ung th biu m, IC50 = 2,0 g/ml), FL (t bo ung
th mng t cung, IC50 = 3,9 g/ml) v Hep-2 (t bo ung th gan, IC50 = 2,4
g/ml) [251].

Hnh V.1.4.4.e. Ph khi lng ESI-MS ca XT4

391

V.1.4.5. Hp cht XT5: cholest-7-en-3-one

Hp cht XT5 phn lp c di dng tinh th mu trng. Ph cng
hng t ht nhn (NMR) ca XT5 tng t nh ca XT3. S trng khp hon
ton v gi tr ph 13C-NMR ti cc v tr tng ng t C-11 n C-27 cho php
khng nh phn cu trc ny ca hai hp cht l ging nhau.
21

26

25
20

18

17

11

27

13

19
1

14
8

10
3
5

Hnh V.1.4.5. Cu trc ha hc ca XT5

Ngoi ra, s trng khp v s liu ph 13C-NMR phn khung steroit ca
XT5 vi cc s liu tng ng ca hp cht c cu trc tng t [245], cng vi
kt qu ph khi lng ESI-MS ti m/z 385,2 [M+H]+ cho php khng nh cu
trc ha hc ca XT5 l cholest-7-en-3-one.
V.1.4.6. Hp cht XT6: saringosterol
Hp cht XT6 cng c phn lp di dng tinh th hnh kim mu trng.
Ph H- v 13C-NMR hon ton tng t nh ca XT3 gi rng hai hp cht
ny c cu trc tng t nhau. Hp cht ny nhn dc di dng mt vt trn
trn TLC v gi tr tch phn trn ph 1H-NMR rt chn. Tuy nhin ti mt vi tn
hiu trn ph 13C-NMR (Bng V.1.4.6) c s tch vch rt st nhau vi t l 1/1
v cng tn hiu. iu ny cho thy y l mt hn hp ng phn c
tng ng v cu trc ha hc rt cao. Chng ti c gng phn tch hn hp
ny bng nhiu phng php sc k khc nhau nhng khng t kt qu mong
mun.
1

29
28
21

18

24
20

26
25

OH
17

11
13

19
1

9
10

27

14
8

HO

Hnh V.1.4.6.a. Cu trc ha hc ca XT6

392

Hnh V.1.4.6.b. Ph 1H-NMR ca XT6

S trng khp hon ton v cc gi tr ph 13C-NMR ti cc v tr tng
ng t C-1 n C-21 gia hai hp cht cho php khng nh hai hp cht ny c
cng cu trc b khung steroit v ch khc nhau v cu trc mch nhnh. S khc
bit d nhn thy nht trn ph 1H-NMR ca XT6 so vi XT3 l s xut hin
thm cc tn hiu proton olefin ti 5,13 (1H, dd, J = 10,5 v 1,5 Hz), 5,18 (1H,
dd, J = 17,5 v 1,5 Hz) v 5,80 (1H, dd, J = 17,5 v 10,5 Hz).
Bng V.1.4.6. Kt qu ph NMR ca XT6
C
1
2
3
4
5
6
7
8
9
10
11
12
13
14

Ca, b
37,27
31,67
71,81
42,31
140,77
121,70
31,91
31,91
50,13
36,52
21,08
39,76
42,34/42,31
56,75

Ha, c

HMBC

dng pic (J = Hz)

(H C)

1,07/1,85
1,50/1,82
3,52 m
2,27
5,35 t, (3,0)
1,52/1,97
1,45 m
0,95 m
1,49 m
1,18/2,20
0,99 m

393

C-2, 3, 5, 10
C-4, 7, 8, 10
C-5, 6

15
16
17
18
19
20
21
22
23
24
25
26
27
28
29

24,28
28,22/28,17
55,83
11,86
19,40
36,10/36,14
18,79
29,10/29,07
34,57/34,79
77,71
35,93
16,47
16,54
142,57/142,49
112,95/112,86

1,05/1,58
1,26/1,82
1,15 m
0,68 s
1,00 s
1,39 m
0,92 d (7,0)
1,03/1,41
1,58/1,62
1,73 m
0,90 d (7,0)
0,87 d (7,0)
5,80 dd (10,5 v 17,5)
5,13 dd (1,5 v
10,5)/5,18 dd (1,5 v
17,5)
a
b
c
in CDCl3, 125 MHz, 500 MHz

C-12, 13, 14, 17

C-1, 5, 9, 10
C-17, 23
C-17, 20, 22

C-24, 26, 27, 28

C-24, 25, 27
C-24, 25, 26
C-24
C-24, 28

Ph 13C-NMR ca XT6 xut hin cc tn hiu ca 29 cacbon trong c

11 nhm metilen, 9 nhm metin, 5 nhm metyl v 4 cacbon bc bn. S khc bit
d nhn thy nht trn ph 13C-NMR ca XT6 so vi XT3 l s xut hin thm
cc tn hiu ca mt ni i ngoi vng v u mch ti 112,9 (t) v 142,5
(d). Tn hiu metilen C-24 mt i v thay vo l s xut hin ca tn hiu
cacbon bc bn gn vi nguyn t xy ti 77,7 (s).

Hnh V.1.4.6.c. Ph 13C-NMR v cc ph DEPT ca XT6

394

Hnh V.1.4.6.d. Ph HSQC ca XT6

Phn tch chi tit cc tng tc trn ph HSQC v HMBC (Bng V.1.4.6 v
Hnh V.1.4.6.f) cng vi kt qu ph khi lng ESI-MS (ti m/z 411,1 [M - H2O
+ H]+, 393,0 [M - 2H2O + H]+) cho php xc nh cu trc ca mch nhnh cng
nh ton b phn t ca XT6 nh c ch ra trn Hnh V.1.4.6.b.

Hnh V.1.4.6.e. Ph HMBC ca XT6

395

29
28

21

18

24
20

26
25

OH
17
11

27

13

19
1

9
10

14
8

3
5

HO

Hnh V.1.4.6.f. Cc tng tc HMBC(HC) chnh ca XT6

iu ny mt ln na c khng nh bi s ph hp hon ton ti cc
gi tr tng ng v cc tnh cht vt l v s liu ph ca XT6 so vi ti liu
c cng b cho saringosterol [252]. Nh vy, cu trc ha hc ca XT6 c
xc nh l saringosterol (di dng hn hp ng phn epimer 24S/24R vi t l
1/1).

Hnh V.1.4.6.g. Ph khi lng (ESI-MS) ca XT6

Hp cht ny ln u tin c phn lp t loi rong nu Sargassum
ringgoldianum (sau tip tc c tm thy nhiu loi rong nu khc), t loi
hi min Xestospongia muta v c thc vt (loi Lesonia nigrescens). Mt iu
ng quan tm l hp cht ny th hin hot tnh khng vi khun lao
Mycobacterium tuberculosis rt mnh vi gi tr nng c ch ti thiu (MIC)
l 0,25 g/ml v y l gi tr MIC nh nht cho cc hp cht thin nhin c
chit tch t thc vt tnh n nm 2001. Gi tr ny tng ng vi gi tr MIC
ca thuc cha lao rifampin. Ngoi ra hp cht ny cn c c thp v tnh
c hiu cao [253]. Theo tc gi Brent R. C. [254], vi lp cht steroit tnh n
nm 2003, ch c 3 hp cht l axit fusidic, saringosterol v endoperoxide c hot
tnh khng vi khun lao vi gi tr MIC 4 g/ml.
396

V.1.4.7. Hp cht XT7: Thymindine

Hp cht XT7 nhn c di dng cht bt mu trng. Cng thc phn t
ca n c xc nh l C10H14N2O5 (M = 242) bng kt qu ph khi lng
(ESI-MS) ti m/z 264,7 [M+Na]+ v 240.8 [M-H]- cng vi cc ph cng hng
t ht nhn (NMR).
O
7

HN 3
2

CH3
5
6

O
5'

HOCH2

O
1'

4'
3'

2'

OH

Hnh V.1.4.7.a. Cu trc ha hc ca XT7

Trn ph 1H-NMR ca XT7 xut hin cc tn hiu c trng ca mt n v
ng 2-deoxy--D-erythro-pentofuranosyl ti 4,41 (1H, m, H-1), 2,24 (1H,
m, H-2), 3,92 (1H, dd, J = 3,5, 6,5 Hz, H-3), 6,29 (1H, m, H-4), 3,75 (2H, m,
H-5).

Hnh V.1.4.7.b. Ph 1H-NMR ca XT7.

397

Bng V.1.4.7. Kt qu ph NMR ca XT7

No
1
2
3
4
5
6
7
1
2
3
4
5

Ca [238]
150,5
164,8
109,5
136,2
12,3
84,9
39,5
70,5
87,3
61,4

Cb, c
152,38
166,42
111,53
138,17
12,42
86,26
41,19
72,21
88,82
62,84

Hc, d
7,82 d (1,0)
1,90 d (1,0)
4,41 m
2,24 m
4,92 dd (4,5, 6,5)
6,29
4,75 m

o trong DMSO, bo trong MeOD, c 125MHz, d500MHz

Hnh V.1.4.7.c. Ph 13C-NMR ca XT7

Ph 13C-NMR ca XT7 xut hin cc tn hiu ca 10 cabon, trong mt
n v ng 2-deoxy--D-erythro-pentofuranosyl [238] c khng nh bi cc
tn hiu cng hng ti 88,84 (d), 86,26 (d), 72,21 (d), 62,84 (t) v 41,19 (t).
Tn hiu cacbon ti 12,42 (q) c trng cho mt nhm metyl gn vi ni i.

398

Hnh V.1.4.7.d. Ph 13C-NMR v cc ph DEPT ca XT7

T cc phn tch nu trn, s liu ph 13C-NMR ca XT7 c so snh vi
s liu c cng b ca hp cht thymidine [238]. S ph hp hon ton
cc gi tr ph tng ng cng vi kt qu ph khi lng cho php khng nh
cu trc ha hc ca XT7 l 1-(2-deoxy--D-erythro-pentofuranosyl)-5-methyl2,4-(1H,3H)-pyrimidinedione (thymidine).

Hnh V.1.4.7.e. Ph ESI-MS ca XT7

V.1.4.8. Hp cht XT8: thymine
Hp cht XT8 cng nhn c di dng cht bt mu trng. Ph 1HNMR ca n hon ton tng t nh ca XT7 ngoi tr s mt i cc tn hiu

399

cng hng ca n v ng. T c th d on XT8 c cu trc trng vi

phn khung ca XT7.

Hnh V.1.4.8.a. Ph 1H-NMR ca XT8

O
7

HN 3
2

CH3
5
6

N
H

Hnh V.1.4.8.b. Cu trc ha hc ca XT8

Phn tch nu trn, cng vi kt qu ph khi lng (ESI-MS) ti m/z
264,7 [M+Na]+ v 240.8 [M-H]- tng ng vi cng thc phn t C10H14N2O5 (M
= 242) cho php khng nh cu trc ha hc ca XT8 l 5-methyl-2,4(1H,3H)pyrimidinedione hay cn c gi l thymine.

Hnh V.1.4.8.c. Ph ESI-MS ca XT8

400

V.1.4.9. Hp cht XT9: chimyl alcohol

Hp cht XT9 nhn c di dng kt tinh mu trng, ph 1H v 13CNMR c trng cho mt glyxerol mono alkyl te. Kt qu ph khi lng (ESIMS) ti m/z: 317,1 [M+H]+ (tng ng vi cng thc phn t C19H40O3, M=316)
v s ph hp hon ton v s liu ph NMR ca XT9 so vi ti liu cng b
[221-222] cho php xc nh cu trc ha hc ca XT9 l chimyl alcohol
(glycerol 1-O-hexadecyl ether).
3

HO

OH

2
1

1'

16'

CH2(CH2)14CH3

Hnh V.1.4.9. Cu trc ha hc ca XT9

V.1.4.10. Hp cht XT10: batilol

Hp cht XT10 cng nhn c di dng tinh th mu trng. Ph 1H v
C-NMR c trng cho mt glyxerol mono alkyl te. Kt qu ph khi lng
(ESI-MS) ti m/z: 345,1 [M+H]+ (tng ng vi cng thc phn t C21H44O3,
M=344) v s ph hp hon ton v s liu ph NMR ca XT10 so vi ti liu
cng b [221-222] cho php xc nh cu trc ha hc ca XT10 l batyl alcohol
(glycerol 1-O-octadecyl ether).
13

HO

OH

1'

18'

CH2(CH2)16CH3

Hnh V.1.4.10. Cu trc ha hc ca XT10

V.1.5. nh gi hot tnh sinh hc ca cc hp cht

V.1.5.1. nh gi hot tnh khng vi sinh vt kim nh
Tt c cc hp cht phn lp c t loi hi min Xestospongia
testudinaria c th nghim v nh gi hot tnh khng vi sinh vt kim
nh trn cc dng vi sinh vt gy bnh tiu biu nht thuc cc nhm vi khun
Gr(-); vi khun Gr(+); nm mc v nm men. nh gi s b hot tnh, chng
ti pha mu nng theo tiu chun sng lc s b v tm cc mu th hin
hot tnh khng vi sinh vt kim nh nng . Cc mu c biu hin dng
tnh bc s b ny c pha long theo cc thang nng thp dn, t 5-10
thang nng tnh gi tr nng ti thiu (MIC) m vi sinh vt b c
ch pht trin gn nh hon ton.
401

Bng IV.1.5.2. Kt qu xc nh gi tr MIC ca XT1 v XT2

Hp cht

Vi khunGr(-)

Nng c ch ti thiu(MIC: g/ml)

Vi khunGr(+)
Nm mc

Nm men

E.coli

P.aeru

B.subti

S.aureus

Asp.niger

F.oxy

S.cere

C.albic

XT1

(-)

50

(-)

50

(-)

50

(-)

(-)

XT2

(-)

50

(-)

50

(-)

50

(-)

(-)

Kt qu th nghim cho thy, ch c hp cht XT1 v XT2 c biu hin

hot tnh khng vi sinh vt kim nh. Chng u khng mnh ba chng vi sinh
vt kim nh l: trc khun m xanh Pseudomonas aeruginosa, khng t cu
vng Staphylococcus aureus v khng nm si Fusarium oxysporum vi gi tr
IC50 thp l 50 g/ml.
Ngy nay, vic s dng cc hp cht c ngun gc thin nhin lm thuc
cha bnh v ang c nhiu nh khoa hc trong v ngoi nc quan tm bi
s a dng v hot tnh sinh hc cng nh c tnh thp ca chng. Hin nay,
nhiu loi thuc c tc dng iu tr cc bnh nhim khun v ang c
nghin cu v ng dng song hu ht cc thuc ny u gp kh khn trong vic
iu tr cc bnh nhim khun do trc khun m xanh Pseudomonas aeruginosa
(thuc nhm vi khun Gram (-) l vi khun gy khng thuc khng sinh mnh v
khng hu ht cc thuc khng sinh th h c) v t cu vng Staphylococcus
aureus (thng gy nhim trng v bi nhim nng ngoi da cng nh mt s c
quan v thng c mt trong cc trng hp nhim trng mu v cng l chng
khng thuc vi nhiu thuc khng sinh). Cc hp cht XT1 v XT2 c hot tnh
mnh khng trc khun m xanh Pseudomonas aeruginosa, khng t cu vng
Staphylococcus aureus v khng nm si Fusarium oxysporum. iu ny to c
s khoa hc vng chc cho cc nghin cu ng dng tip theo nhm to ra cc
dc phm c tc dng phng v iu tr cc bnh nhim khun da trn vic
khai thc ngun dc liu bin qu gi, sn c trong nc vo phc v cuc sng.
Ngoi ra, tuy khng c biu hin hot tnh trn 6 chng vi sinh vt kim
nh nghin cu, hp cht XT6 (saringosterol) li c hot tnh khng vi khun
lao Mycobacterium tuberculosis rt mnh vi gi tr nng c ch ti thiu
(MIC) l 0,25 g/ml v y l gi tr MIC nh nht cho cc hp cht thin nhin
c chit tch t thc vt tnh n nm 2001. Gi tr ny tng ng vi gi tr
MIC ca thuc cha lao rifampin. iu ny chng t XT6 c kh nng khng vi
khun lao vi tnh c hiu rt cao. Cc nghin cu cng b cn chng minh
hp cht ny c c tnh rt thp [253]. Theo tc gi Brent R. C. [254], vi lp
cht steroit tnh n nm 2003, ch c 3 hp cht l axit fusidic, saringosterol v
endoperoxide c hot tnh khng vi khun lao vi gi tr MIC 4 g/ml. iu
ny t c s khoa hc cho hng nghin cu su rng hn v hp cht XT6
cng nh loi hi min Xestospongia testudinaria theo nh hng hot tnh
khng vi khun lao nhm nghin cu v pht trin cc ch phm thuc tr bnh
lao th h mi, phc v cng tc chm sc sc khe cng ng.

402

V.1.5.2. nh gi hot tnh gy c t bo

Tt c cc hp cht phn lp c t loi hi min X. testudinaria c
tin hnh nh gi hot tnh khng vi sinh vt kim nh. Kt qu th nghim cho
thy, ch c hp cht XT4 biu hin hot tnh trn 03 dng t bo th nghim
l KB, FL v Hep-G2 vi gi tr IC50 ln lt l 2,0, 3,93 v 2,4 g/ml.
Bng V.1.5.1. Kt qu nh gi hot tnh gy c t bo ca cc hp cht
Hp cht
DMSO
Ellipthine
XT1
XT2
XT3
XT4
XT5
XT6
XT7
XT8
XT9
XT10

Gi tr CS% (Cell survival)

KB
1000,0
1,071,07
58,92,1
67,11,5
79,22,3
32,31,7
69,13,1
74,62,5
59,80,9
79,62,1
65,73,4
79,62,5

FL
1000,0
1,520,00
84,12,2
71,91,4
92,33,1
44,61,5
75,93,1
88,52,4
79,11,5
74,93,4
78,13,6
74,91,4

Hep-G2
100,00,0
1,340,63
86,70,8
55,71,6
77,32,9
36,70,8
56,70,9
65,32,7
56,4 2,1
56,71,3
66,21,7
56,70,8

IC50 (g/ml)
KB
0.22
(-)
(-)
(-)
2,00
(-)
(-)
(-)
(-)
(-)
(-)

FL
0.11
(-)
(-)
(-)
3,93
(-)
(-)
(-)
(-)
(-)
(-)

Hep-G2
0.19
(-)
(-)
(-)
2,40
(-)
(-)
(-)
(-)
(-)
(-)

V.1.6. Cc cng trnh cng b

T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc
loi hi min Xestospongia testudinaria, chng ti bin son v gi ng ti 02
cng trnh trn tp ch chuyn ngnh quc gia v 01 bng c quyn pht minh
sng ch ti Cc S hu Tr tu Vit Nam l:
(1) Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn
Kim v Chu Vn Minh, Nghin cu thnh phn ha hc ca loi hi min
Xestospongia testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v Cng ngh,
2007, 45(3), 43-50
(2) Chu Vn Minh, Phan Vn Kim Trn Anh Tun v Nguyn Xun Cng, Two new
brominated fatty acids from the sponge Xestospongia testudinaria living in Vietnamese
sea. Advance in Natural Science, 2008, 9(2), 167-171.
(3) Chu Vn Minh, Nguyn Xun Cng, Phan Vn Kim, L Mai Hng v Cng
Thung, Hp cht axt 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic
v phng php chit hp cht ny t loi hi min xestospongia testudinaria. S hu
Tr tu Vit Nam, 2008 ( chp nhn n v ang thm nh ni dung).

403

tng hp cc cht phn lp c t loi hi min

xestospongia testudinaria
6

COOH

9
17
15

11

Br
19

XT1: axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene7,9,17-triynoic (cht mi)

8

COOH

9
11

19
13

17

22

Br

21

XT2: axit 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19-triynoic

(cht mi)
s
18

21

17

14

11
29

10

21

1
24

20

OH
21

24

HO

27

26

20

XT3: Cholesterol s =

10

25

XT6: Saringosterol s =

14

26

27

13

19

28

HO

20

18

13

19
1

26

25

17

11

XT4: 5,8-epidioxycholest-6-en-3-ol

25

27

21
25

26

20

18

17

11

27

HN3

HOCH2
O

13

19
1

14

O
3

XT7

R=

XT8

R= H

XT7: Thimidine
XT8: Thimine

XT5: cholest-7-en-3-one
3

HO

OH

XT9: Chimyl alcohol

1
1'

CH2(CH2)nCH3

404

XT10:

Batyl alcohol

3'

2'

OH

1'

4'

10

5'

CH3

n = 14
n = 16

V.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca

loi hi min Gellius varius (chanlinidae)
V.2.1. M u
Mu hi min Gellius varius c thu thp vo thng 11 nm 2006 ti Hi
Phng v thng 01 nm 2007 ti Qung Ninh v c TS. Cng Thung v cc
cng s, Vin Ti nguyn v Mi trng Bin, Vin KH&CN Vit Nam gim
nh tn khoa hc.
- M t: dng cnh, mu xanh lc
m khi cn sng, khi cht hoc khi c
nh bng ho cht th chuyn mu
xanh en, chng to thnh khm sng
trn cc khi san h hoc bm trn
nn y cng

Hi min: Gellius varius

Gellius l mt chi hi min sinh sng kh ph bin ti cc vng bin Vit
Nam. Hin nay nc ta hon ton cha c cng trnh khoa hc no c cng
b v thnh phn ha hc v hot tnh sinh hc ca cc loi thuc chi ny. Trn
th gii hin cng c rt t cc cng trnh cng b v chi ny v mi c 11 hp
cht c phn lp v xc nh cu trc. Nm 1994, nhm nghin cu ca tc gi
Bifulco G. cng b s phn lp v xc nh cu trc ca 2 hp cht di dng hn
hp ng phn lp th l ()-gelliusine A v B t chi Gellius [255]. Nm 1995,
nhm nghin cu ny li cng b thm 6 hp cht l: ()-gelliusine C-F v 2,2bis-(6-bromo-3-indolyl)-ethylamine t chi ny [256]. Gn y nht (2001), 4
hp cht sterol l gelliusterol A-D c phn lp t chi ny [257].
Di y, chng ti gii thiu mt s hp cht quan trng c phn
lp t chi Gellius.
Br

Br

NH2

NH2

NH2

HN
HO

NH2

NH2

HO
HN
N
H
NH2

N
H
N
H

N
H

()-gelliusine C

()-gelliusine A v B
405

Br

Br

NH2

NH2
NH2

HO

NH2

HN

N
H

HO

N
H

N
H

()-gelliusine E

()-gelliusine D
NH2

Br

NH2

Br

Br

NH2

HN

N
H
N
H

N
H

Br

()-gelliusine F

Br

2,2-bis-(6-bromo-3-indolyl)ethylamine

H
H

HO
OH

HO

gelliusterol A

gelliusterol B

OH
H

HO

HO

gelliusterol C
gelliusterol D

406

V.2.2. Phn lp cc hp cht

Mu hi min ti (3 kg) c xay nh, sau chit vi methanol ba ln,
mi ln 72 gi. Cc dch chit methanol c gp li v ct loi dung mi di
p sut gim thu, ha vo nc ri tin hnh loi mui trn ct Dianion vi dung
mi ra gii l nc. Dng MeOH ra gii thu hi cht thu c 15 gam dch
c. Dch c ny c ho tan vo 01 lt nc ct sau c chit ln lt bng
n-hexan, cloroform, v BuOH (mi ln 02 lit) thu c cc cn dch tng ng:
hexan (2 g), cloroform (4 g), BuOH (6 g) v nc (3 g) (S V.2.2.a).
Cn chit n-hexan: GH (2 g) c tin hnh phn tch trn sc k ct lp
li nhiu ln vi cc h dung mi thch hp (S V.2.2.b) thu c cc hp cht
HM-8, 9 v 10.

Hi min Gellius varius ti

(3 kg)

- Chit MeOH, ct loi dung mi di p

sut gim
- Loi mui bng cch chy qua ct
Dianion vi dung mi ra gii l nc ct,
sau thu hi cht bng MeOH.
B sung nc

GM - Cn chit MeOH (15g)

B sung n-hexan

n-Hexan: nc 1/1

Lp nc
B sung CHCl3

GH - Cn n-hexan (2 g)

CHCl3: nc 1/1

Lp nc
B sung BuOH

GC - Cn CHCl3 (4 g)

BuOH: nc 1/1
lp nc

GB - Cn BuOH (6 g)

GN - Cn nc (3 g)

S V.2.2.a: S chit phn on mu Hi min

407

GH - (2 g)
Silica gel CC
Hexan/Axeton 100/0-0/100

GH1 (250mg)

GH2 (375mg)

GH3 (225mg)

GH4 (600mg)

GH5 (550g)

Silica gel CC
n-hexan/axeton/ EtOAc:
50/1/1

Silica gel CC
n-hexan/Etylaxetat
40/1
HM-8
(10mg)

HM-9
(100mg)

HM-10
(5mg)

S V.2.2.b: S phn lp cc hp cht t cn chit n-hexan

Cn chit clorofrom: GC (4 g) c tin hnh chy ct th trn sc k ct
pha thng gradient nng metanol trong cloroform thu c 5 phn on nh
hn l GC1-5. Cc phn on GC1, GC3 v GC5 c tip tc phn tch trn sc
k ct pha thng vi h dung mi ra gii thch hp thu c cc hp cht HM4, 5, 6 v 7 nh c ch ra trn S V.2.2.c. Bng cc phng php tng t
(S V.2.2.d), cc hp cht HM-1, 2, v 3 c phn lp t cn chit BuOH.
GC (4 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

GC1 (1g)

GC2 (600mg)

GC4 (200mg)

Silica gel CC
n-hexan/CHCl3/axeton 4/1/0,1

Silica gel CC
n-hexan/axeton: 7/1

HM-4
(7mg)

GC3 (1.3g)

HM-5
(12mg)

HM-6
(8mg)

GC5 (900mg)

Silica gel CC
CHCl3/MeOH
20/1
HM-7
(9mg)

S V.2.2.c: S phn lp cc hp cht t cn chit cloroform

408

GB - (6 g)

Silica gel CC
Cloroform/MeOH/H2O: 5/1/0 - 2/1/0.2

GB1 (1.0 g)

GB2 (2.5 g)

Silica gel CC
CHCl3/MeOH/H2O
5/1/0.1
HM-1
(12mg)

GB3 (1.5 g)

GB4 (600mg)

Silica gel CC
CHCl3/MeOH/H2O
4/1/0.1
HM-2
(90mg)

GB5 (1.4 g)

Silica gel CC
CHCl3/MeOH/H2O
3/1/0.2
HM-3
(15mg)

S V.2.2.d: S phn lp cc hp cht t cn chit butanon

V.2.3. Hng s vt l v d kin ph ca cc hp cht
V.2.3.1. Hp cht HM1: 1-O-palmitoyl-3-O-[-D-galactopyranosyl(16)--Dgalactopyranosyl]-sn-glycerol
25
Cht bt v nh hnh mu trng, []D +56,5 (MeOH, c 0.5), ph khi lng
ESI-MS: m/z 653,3 [M-H]+, 677,3 [M+Na]+, 515,2 [M+Na-162]+, 689,4 [M+2H2O-H]+,
(C31H58O14, M = 654).
1
H-NMR (500 MHz, CD3OD + CDCl3) H: 4,13 (2H, H-1), 3,98 (1H, m, H-2),
3,72 (1H, H-3a), 3,81(1H, H-3b), 4,25 (1H, d, J = 7,5 Hz, H-1), 3,57 (1H, H-2), 3,53
(1H, H-3), 3,90 (1H, H-4), 3,69 (1H, H-5), 3,68 (1H, H-6a), 3,90 (1H, H-6b), 4,91
(1H, d, J = 3,5 Hz, H-1) 3,74 (1H, H-2), 3,79 (1H, H-3), 3,94 (1H, H-4), 3,83 (1H,
H-5), 3,75 (2H, H-6), 2,35 (2H, t, J = 9,0 Hz, H-2), 1,62 (2H, t, J = 7,0 Hz, H-3),
1,29 (20H, br s, H-4-13) 1,29 (2H, br s, H-14), 1,29 (2H, br s, H-15), 0,88 (3H,
t, J = 7,0 Hz, H-16) ppm.
13
C-NMR (125 MHz, CD3OD + CDCl3) C: 65,35 (t, C-1), 68,78 (d, C-2), 71,92
(t, C-3), 104,13 (d, C-1), 71,56 (d, C-2), 73,51 (d, C-3), 68,64 (d, C-4), 73,31 (d, C5), 66,58 (t, C-6), 99,40 (d, C-1), 70,39 (d, C-2), 69,25 (d, C-3), 69,96 (d, C-4),
70,97 (d, C-5), 61,96 (t, C-6), 174,75 (s, C-1), 34,35 (t, C-2), 25,10 (t, C-3),
29,38 - 29,90 (t, C-4-13), 32,14 (t, C-14), 22,88 (t, C-15), 14,17 (q, C-16) ppm.
V.2.3.2. Hp cht HM2: (2S)1,2-Di-O-Palmitoyl-3-O-(6-sulfo--D-quinovopyranosyl)
Glycerol
25
Cht bt v nh hnh mu trng, []D +42,5 (MeOH, c 1.0), ph khi lng
ESI-MS: m/z 793,6 [M-H]+ (C41H78O12S, M = 794).
1
H-NMR (500 MHz, CDCl3+CD3OD) H: 4,20 (1H, dd, J = 7,0, 12,0 Hz, H-1a),
4,43 (1H, dd, J = 3,0, 12,0 Hz, H-1b), 5,31 (1H, m, H-2), 3,62 (1H, dd, J = 6,5, 11, 0 Hz,
H-3a), 3,98 (1H, dd, J = 5,0, 11,0 Hz, H-3b), 4,83 (1H, d, J = 3,5 Hz, H-1), 3,51 (1H,
dd, J = 3,5, 9,5 Hz, H-2), 3,65 (1H, dd, J = 9,0, 9,5 Hz, H-3), 3,38 (1H, dd, J = 9,0, 9,5
Hz, H-4), 4,02(1H, m, H-5), 3,14 (1H, dd, J = 6,0, 14,5 Hz, H-6a), 3,33 (1H, dd, J =

409

4,0, 14,5 Hz, H-6b), 2,33 (2H, m, H-2), 1,61 (2H, m, H-3), 1,30 (20H, br s, H-413), 1,30 (2H, br s, H-14), 1,30 (2H, br s, H-15), 0,89 (3H, t, J = 6,5Hz, H-16), 2,33
(2H, m, H-2), 1,61 (2H, m, H-3), 1,30 (20H, br s, H-4-13), 1,30 (2H, br s, H14) ppm.
13
C-NMR (125 MHz, CDCl3+CD3OD) C: 63,45 (t, C-1), 70,84 (d, C-2), 66,68 (t,
C-3), 99,28 (d, C-1), 72,22 (d, C-2), 73,97 (d, C-3), 73,58 (d, C-4), 68,77 (d, C-5),
53,59 (d, C-6), 174,46 (s, C-1), 34,66 (t, C-2), 25,22 (t, C-3), 29,45 - 29,94 (t, C-413), 32,17 (t, C-14), 22,87 (t, C-15), 14,11 (q, C-16), 174,27 (s, C-1), 34,46 (t, C2), 25,18 (t, C-3), 29,45 - 29,94 (t, C-4-13), 32,17 (t, C-14), 22,87 (t, C-15),
14,11 (q, C-16) ppm.
V.2.3.3. Hp cht HM3: Mui natri ca (2S)1-O-Hexadecanoyl-3-O-(6-sulfo--Dquinovopyranosyl) Glycerol
Cht bt v nh hnh mu trng, ph khi lng ESI-MS: m/z 555,3 [M - Na]+,
601,2 [M + Na]+, (C25H47NaO11S, M = 578).
1
H-NMR (500 MHz, CD3OD) H: 4,12 (1H, H-1a), 4,22 (1H, dd, J = 6,5, 13,5
Hz, H-1b), 4,11 (1H, m, H-2), 4,08 (1H, H-3a), 3,42 (1H, H-3b), 4,81 (1H, d, J = 3,5Hz,
H-1), 3,43 (1H, H-2), 3,68 (1H, t, J = 9,0Hz, H-3), 3,12 (1H, t, J = 9,0Hz, H-4), 4,12
(1H, H-5), 2,95 (1h, dd, J = 9,5, 14,5Hz, H-6a), 3,38 (1H, H-6b), 2,39 (1H, m, H-2),
1,63 (1H, m, H-3), 1,30 (20H, br s, H-4-13), 1,30 (2H, br s, H-14), 1,30 (2H, br s,
H-15), 0,93 (3H, t, J = 6,5Hz, H-16) ppm.
13
C-NMR (125 MHz, CD3OD) C: 66,53 (t, C-1), 69,79 (d, C-2), 70,49 (t, C-3),
100,12 (d, C-1), 73,59 (d, C-2), 75,07 (d, C-3), 74,88 (d, C-4), 69,79 (d, C-5), 54,19
(d, C-6), 175,59 (s, C-1), 34,93 (t, C-2), 25,97 (t, C-3), 30,22 - 30,76 (t, C-4-13),
33,04 (t, C-14), 23,70 (t, C-15), 14,42 (q, C-16) ppm.
V.2.3.4. Hp cht HM4: Cholesterol
Cht kt tinh mu trng, nhit nng chy 148-149 oC, C27H46O, M=386.
1
H-NMR (500 MHz, CDCl3) : 3,52 (1H, m, H-3), 5,35 (1H, br d, J = 5,0 Hz, H6), 1,00 (3H, s, H-19), 0,91 (3H, s, H-21), 0,86 (6H, d, J = 6.5 Hz), H-26, 27) and 0.68
(3H, s, H-18).
13
C-NMR (125 MHz, CDCl3) : 37.2 (t, C-1), 31.6 (t, C-2), 71.8 (d, C-3), 42,2 (t,
C-4), 140,7 (s, C-5), 121,7 (d, C-6), 31,8 (t, C-7), 31,8 (d, C-8), 50,1 (d, C-9), 36,5 (s, C10), 21,1 (t, C-11), 39,8 (t, C-12), 42,3 (s, C-13), 56,7 (d, C-14), 24,3 (t, C-15), 28,3 (t,
C-16), 56,1 (d, C-17), 11,9 (q, C-18), 19,4 (q, C-19), 35,7 (d, C-20), 18,7 (q, C-21), 36,1
(t, C-22), 23,8 (t, C-23), 39,5 (t, C-24), 28,0 (d, C-25), 22,5 (q, C-26), 22,8 (q, C-27).
V.2.3.5. Hp cht HM5: 5,8-Epidioxycholest-6-en-3-ol
25
Tinh th hnh kim mu trng, []D - 30 (CHCl3, c 0,5), nhit nng chy 102105oC, ph khi lng ESI-MS: m/z 439 [M+Na]+, (C27H44O3, M = 416).
1
H-NMR (500 MHz, CDCl3) H: 1,96 (1H, H-1a), 2,02 (1H, H-1b), 1,54 (1H, H2a), 1,81 (1H, m, H-2b), 3,80 (1H, H-3), 1,95 (1H, H-3a), 2,15 (1H, H-3b), 6,30 (1H, d,
J = 8,5Hz, H-6), 6,55 (1H, d, J = 8,5Hz, H-7), 1,52 (1H, H-9), 1,06 (1H, m, H-11a),
1,59 (1H, H-11b), 1,22 (2H, H-12), 1,58 (1H, d, J = 3,5Hz, H-14), 1,21 (1H, H-15a),
1,52 (1H, H-15b), 1,45 (2H, H-16), 1,20 (1H, H-17), 0,86 (3H, s, H-18), 0,92 (3H, s, H19), 1,60 (1H, H-20), 0,96 (3H, d, J = 6,5Hz, H-21), 1,73 (2H, m, H-22), 1,32 (2H, m,

410

H-23), 1,24 (2H, H-24), 1,45 (1H, H-25), 0,90 (3H, d, J = 6,5Hz, H-26), 0,90 (3H, d, J
= 6,5Hz, H-27) ppm.
13
C-NMR (125 MHz, CDCl3) C: 40,65 (t, C-1), 30,87 (t, C-2), 66,98 (d, C-3),
37,17 (t, C-4), 83,49 (s, C-5), 136,80 (d, C-6), 131,75 (d, C-7), 80,78 (s, C-8), 52,74 (d,
C-9), 37,78 (s, C-10), 21,58 (t, C-11), 40,82 (t, C-12), 45,95 (s, C-13), 52,74 (d, C-14),
24,41 (t, C-15), 29,25 (t, C-16), 57,81 (d, C-17), 13,04 (q, C-18), 19,07 (q, C-19), 36,52
(d, C-20), 18,59 (q, C-21), 37,16 (t, C-22), 24,90 (t, C-23), 40,65 (t, C-24), 29,25 (d, C25), 22,92 (q, C-26), 23,17 (q, C-27) ppm.
V.2.3.6. Hp cht HM6: Cholest-7-ene-3,5,6-triol
25
Tinh th hnh kim mu trng, [ ]D -20.8 (MeOH, c 0.5), ESI-MS: m/z 401,2 [M+
+
H2O+H] , 383,2 [M-2H2O+H] , 365,2 [M-3H2O+H]+, (C27H46O3, M = 418).
1
H-NMR (500 MHz, CDCl3) H: 1,45 (1H, H-1a), 1,85 (1H, H-1b), 1,55 (1H, H2a), 1,62 (1H, H-2b), 4,09 (1H, m, H-3), 1,79 (1H, dd, J = 5,0, 13,0 Hz, H-4a), 2,10
(1H, dd, J = 13,0, 13,0 Hz, H-4b), 3,63 (1H, br s, H-6), 5,37 (1H, H-7), 1,96 (1H, H-9),
1,59 (2H, H-11), 1,32 (1H, H-12a), 1,80 (1H, H-12b), 1,91 (1H, H-14), 1,35 (1H, H15a), 1,60 (1H, H-15b), 1,32 (1H, H-16a), 1,92 (1H, H-16b), 1,26 (1H, H-17), 0,60 (3H,
s, H-18), 1,09 (3H, s, H-19), 1,39 (1H, H-20), 0,94 (3H, d, J = 6,5Hz, H-21), 1,11 (1H,
H-22a), 1,39 (1H, H-22b), 1,55 (2H, H-23), 1,53 (2H, H-24), 1,52 (1H, H-25), 0,88 (3H,
d, J = 6,5Hz, H-26), 0,89 (3H, d, J = 6,5Hz, H-27) ppm.
13
C-NMR (125 MHz, CDCl3) C: 32,98 (t, C-1), 30,87 (t, C-2), 67,74 (d, C-3),
39,49 (t, C-4), 76,01 (s, C-5), 73,69 (d, C-6), 117,56 (d, C-7), 144,05 (s, C-8), 43,49 (d,
C-9), 37,15 (s, C-10), 22,08 (t, C-11), 39,49 (t, C-12), 43,89 (s, C-13), 54,72 (d, C-14),
22,94 (t, C-15), 27,77 (t, C-16), 56,25 (d, C-17), 12,10 (q, C-18), 18,84 (q, C-19), 36, 07
(d, C-20), 18,84 (q, C-21), 36,14 (t, C-22), 23,93 (t, C-23), 39,37 (t, C-24), 28,02 (d, C25), 22,56 (q, C-26), 22,82 (q, C-27) ppm.
V.2.3.7. Hp cht HM7: 6-Methoxycholest-7-ene-3,5-diol
25
Tinh th hnh kim mu trng, []D -55.6 (c, 0.19 CHCl3), ESI-MS: m/z 415,3 [M+
+
H2O+H] , 397,2 [M-2H2O+H] , 383,3 [M-2H2O-Me+H]+, 365,3 [M-2H2O-MeH2O+H]+, 431,1 [M-H]+, (C28H48O3, M = 432).
1
H-NMR (500 MHz, CDCl3) H: 1,50 (1H, H-1a), 1,90 (1H, H-1b), 1,60 (1H, H2a), 1,68 (1H, H-2b), 4,05 (1H, m, H-3), 1,85 (1H, dd, J = 5,0, 13,0 Hz, H-4a), 2,15
(1H, dd, J = 13,0, 13,0 Hz, H-4b), 3,17 (br d, 5,0) 5,41 (1H, m, H-7), 2,05 (1H, H-9),
1,59 (2H, H-11), 1,34 (1H, H-12a), 1,90 (1H, H-12b), 1,91 (1H, H-14), 1,39 (1H, H15a), 1,65 (1H, H-15b), 1,32 (1H, H-16a), 1,92 (1H, H-16b), 1,26 (1H, H-17), 0,60 (3H,
s, H-18), 1,00 (3H, s, H-19), 1,42 (1H, H-20), 0,93 (3H, d, J = 6,5Hz, H-21), 1,22 (1H,
H-22a), 1,43 (1H, H-22b), 1,57 (2H, H-23), 1,53 (2H, H-24), 1,52 (1H, H-25), 0,88 (3H,
d, J = 6,5Hz, H-26), 0,87 (3H, d, J = 6,5Hz, H-27), 3,40 (3H, s, OCH3) ppm.
13
C-NMR (125 MHz, CDCl3) C: 32,78 (t, C-1), 30,92 (t, C-2), 67,86 (d, C-3),
39,51 (t, C-4), 76,36 (s, C-5), 82,48 (d, C-6), 114,97 (d, C-7), 143,75 (s, C-8), 43,92 (d,
C-9), 37,27 (s, C-10), 22,19 (t, C-11), 39,51 (t, C-12), 43,99 (s, C-13), 54,93 (d, C-14),
22,94 (t, C-15), 27,81 (t, C-16), 56,25 (d, C-17), 12,07 (q, C-18), 18,36 (q, C-19), 36,16
(d, C-20), 18,84 (q, C-21), 36,10 (t, C-22), 23,93 (t, C-23), 39,62 (t, C-24), 28,02 (d, C25), 22,56 (q, C-26), 22,82 (q, C-27), 58,27 (q, -OCH3) ppm.

411

V.2.3.8. Hp cht HM8: Metyl este ca axit (5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic

Cht du khng mu, ph khi lng ESI-MS: m/z 317,1 [M+H]+, 315,0 [M-H]+
(C21H32O2, M = 316).
1
H-NMR (500 MHz, CDCl3) H: 2,32 (2H, t, J = 7,5Hz, H-2), 1,71 (2H, H-3),
2,10 (2H, H-4, 19), 5,38 (10H, m, H-5, 6, 8, 9, 11, 12, 14, 15, 17, 18), 2,83 (8H, m, H-7,
10, 13, 16), 0,98 (3H, t, J = 7,5Hz, H-20), 3,67 (3H, s, OCH3) ppm.
13
C-NMR (125 MHz, CDCl3) C: 174,06 (s, C-1), 33,47 (t, C-2), 26,58 (t, C-3), 128,97
(d), 128,87 (d), 128,59 (d), 128,28 (d), 128,25 (d), 128,15 (d), 128,13 (d), 127,90 (d),
127,04 (d), 25,65 (t), 25,56 (t), 24,81 (t), 132,06 (d, C-18), 20,57 (t, C-19), 14,27 (q, C20), 51,48 (q, OCH3) ppm.
V.2.3.9. Hp cht HM9: Phytol
Cht du khng mu, (C20H40O, M = 296).
1
H-NMR (500 MHz, CDCl3) H: 4,14 (2H, d, J = 7,0Hz, H-1), 5,40 (1H, d, J =
7,0Hz, H-2), 1,99 (2H, t, J = 7,5Hz, H-4), 1,02-1,56 (19H, H-5-15), 0,86 (12H, H-1719), 1,67 (3H, s, H-20) ppm.
13
C-NMR (125 MHz, CDCl3) C: 59,40 (t, C-1), 123,17 (d, C-2), 140,20 (s, C-3),
39,90 (t, C-4), 25,17 (t, C-5), 36,70 (t, C-6), 32,82 (d, C-7), 37,46 (t, C-8), 24,49 (t, C-9),
37,32 (t, C-10), 32,72 (d, C-11), 37,39 (t, C-12), 24,82 (t, C-13), 39,40 (t, C-14), 27,99
(d, C-15), 22,64 (q, C-16), 22,73 (q, C-17), 19,73 (q, C-18), 19,77 (q, C-19), 16,18 (q,
C-20) ppm.
V.2.3.10. Hp cht HM10: Glycerol 1-(5Z,8Z,11Z,14Z-eicosatetraenoate)
Cht du khng mu, ph khi lng ESI-MS: m/z 379,2 [M+H]+, 361,2 [MH2O+H]+, 347,2 [M-H2O-CH2+H]+, 329,1 [M- H2O-CH2-H2O+H]+, 287,2 [MGlycerol+H]+, 377,1 [M-H]+; C23H38O4 (M=378).
1
H-NMR (500 MHz, CDCl3) H: 3,72 (1H, dd, J = 4,0, 11,5Hz, H-1a), 3,62 (1H,
dd, J = 5,5, 11,5Hz, H-1b), 3,95 (1H, m, H-2), 4,23 (1H, dd, J = 4,5, 11,5Hz, H-3a),
4,17 (1H, dd, J = 6,5, 11,5Hz, H-3b), 2,39 (2H, t, J = 7,5Hz, H-2), 1,74 (2H, q, J =
7,5Hz, H-3), 2,85 (6H, m, H-4, 7, 10), 5,37 (8H,m, H-5, 6, 8, 9, 11, 12, 14,
15), 2,08 (2H, m, H-16), 1,33 (6H, m, H-17, 18, 19), 0,91 (3H, t, J = 7,0Hz, H-20)
ppm.
13
C-NMR (125 MHz, CDCl3) C: 63,36 (t, C-1), 70,28 (d, C-2), 65,27 (t, C-3),
174,06 (s, C-1), 33,51 (t, C-2), 29,34 (t, C-3), 24,74 (t),129,07 (d),127,86 (d), 26,53
(t), 128,76 (d), 128,65 (d), 25,65 (t), 128,32 (d), 128,11 (d), 25,64 (t), 127,55 (d), 130,54
(d, C-15), 27,24 (t, C-16), 29,34 (t, C-17), 31,53 (t, C-18), 22,58 (t, C-19), 14,07 (q,
C-20) ppm.

412

V.2.4. Xc nh cu trc ha hc ca cc hp cht

V.2.4.1. Hp cht HM1: 1-O-palmitoyl-3-O-[-D-galactopyranosyl(16)--Dgalactopyranosyl]-sn-glycerol
Hp cht HM1 nhn c di dng bt v nh hnh mu trng. Cng
thc phn t ca HM1 c xc nh l C31H58O14 bng cc d kin ph khi
(m/z 653,3 [M-H]+, 677,3 [M+Na]+) v ph cng hng t ht nhn. Pc ion phn
mnh ti m/z 515,2 [M-162+Na]+ trn ph khi lng th hin s phn ct mt
ng 6 cacbon ca HM1.

Hnh V.2.4.1.a. Ph 1H-NMR ca HM1

Trn ph 1H-NMR xut hin cc tn hiu proton ca: mt nhm metyl ti
0,88 (3H, t, J = 7,0Hz); cc nhm oxymetilen v oxymetin ti 3,5 - 4.3; v mt
mch cacbon no di ti 1,29 (24H, br s). Cc du hiu ny c trng cho mt
glycolipid cha mt axit bo no. Cc tn hiu proton anomer ti 4,25 (1H, d, J =
7.5Hz) v 4,91 (1H, d, J = 3,5) khng nh s tn ti ca 2 lin kt ng: mt
v mt .
HO
OH
4"

6"
5"

O
1"

OH
3"

2"

OH OH
4'

6'
5'

OH
3'

O
1'

2'

OH

CH2
2

HC

OH

CH2 O

1'"

16'"

(CH2)14CH3

Hnh V.2.4.1.b: Cu trc ha hc ca HM1

413

Trn ph 13C-NMR xut hin tn hiu ca mt nhm metyl ti 14,17;

mt tn hiu ca nhm cacbonyl ti 174,75 khng nh s tn ti ca mt nhnh
axit bo duy nht. Cc tn hiu ca cc nhm metilen xut hin ti 29,38-29,90.
Hai tn hiu cc bon anomer xut hin ti 99,40 v 104,1V.2.4. Cc tn hiu
cacbon ca cc n v ng xut hin ti 61,9 - 73,6. Cc chuyn dch ha
hc tng i thp thp gi cho s c mt ca cc n v ng galactose.
chuyn dch ha hc cao ca cacbon C-6 ( 66,58 so vi 61,96) cho php khng
nh n v ng th 2 c ni vo v tr C-6 ca n v ng th nht.

Hnh V.2.4.1.c. Ph 13C-NMR ca HM1

Bng V.2.4.1: Kt qu ph cng hng t ht nhn ca HM1
C
1
2

C [258]
66,6 t
69,7 d

Ca,b
65,35 t
68,78 d

72,1 t

71,92 t

1
2
3
4
5

105,3 d
72,6 d
74,7 d
70,1 d
74,6 d

104,13 d
71,56 d
73,51 d
68,64 d
73,31 d

67,8 t

66,58 t

1
2
3
4
5

100,5 d
71,5 d
70,2 d
71,0 d
72,5 d

99,40 d
70,39 d
69,25 d
69,96 d
70,97 d

Ha,c (J, Hz)

4,13 *
3,98 (m)
3,72
3,81
4,25 (d, 7,5)
3,57
3,53
3,90
3,69
3,68
3,90
4,91 (d, 3,5)
3,74
3,79
3,94
3,83
414

HMBC (H to C)
C-2, 3, 1
C-1
C-3

C-1
C-6

6
1
2
3
413
14
15
16
a

62,7 t
175,5 s
34,9 t
26,0 t
30,2 30,8 t
33,0 t
23,7 t
14,4 q

61,96 t
174,75 s
34,35 t
25,10 t
29,38 29,90 t
32,14 t
22,88 t
14,17 q

3,75
2,35 (t, 9,0)
1,62 (t, 7,0)

C-1, 3
C-1, 2

1,29 (br s)
1,29 (br s)
1,29 (br s)
0,88 (t, 7,0)

C-14, 15

o trong CDCl3+MeOD, b 125 MHz, c 500 MHz

Hnh V.2.4.1.d. Ph 13C-NMR v cc ph DEPT ca HM1

xc nh cu trc tng th ca HM1, cc ph 2D-NMR c tin
hnh. Ph HSQC cho php xc nh cc proton gn trc tip vi cacbon tng
ng (bng V.2.4.1). Trn ph HMBC, s xut hin tn hiu tng tc ca H-1
vi C-6 v H-6 vi C-1 mt ln na khng nh v tr lin kt ca hai n v
ng. Chui 2 n v ng c xc nh gn vo v tr C-3 bi s xut hin tn
hiu tng tc ca proton anomer H-1 vi C-3 v ca H-3 vi cacbon anomer C1. Tn hiu tng tc ca H-1 vi nhm cacbonyl (C-1) cho php khng nh v
tr lin kt ca nhnh axit bo ti v tr C-1.

415

Hnh V.2.4.1.e. Ph HSQC ca HM1

416

Hnh V.2.4.1.f. Ph HMBC ca HM1

417

HO
OH
4"

6"
5"

O
1"

OH
3"

2"

OH OH
4'

6'
5'

OH
3'

O
1'

2'

OH

CH2
2

HC

OH

CH2 O

1'"

16'"

CH2CH2(CH2)12CH3

Hnh V.2.4.1.g. Mt s tng tc HMBC chnh ca HM1

Cc kt qu phn tch nu trn c mang so snh vi cc d kin ph ca

hp
cht
1-O-palmitoyl-3-O-[-D-galactopyranosyl(16)--Dgalactopyranosyl]-sn-glycerol [258]. S trng khp hon ton v cc gi tr ph
tng ng cho php khng nh HM1 l 1-O-palmitoyl-3-O-[-Dgalactopyranosyl(16)--D-galactopyranosyl]-sn-glycerol. Hp cht ny
c phn lp t cc loi to lc Ulva pertusa v Enteromorpha prolifera. Tuy
nhin y l ln u tin n c phn lp t cc loi hi min thuc chi Gellius.

Hnh V.2.4.1.h. Ph ESI-MS ca HM1

418

V.2.4.2.
Hp
cht
HM2:
quinovopyranosyl) Glycerol

(2S)1,2-Di-O-Palmitoyl-3-O-(6-sulfo--D-

Hp cht HM2 nhn c di dng cht bt mu trng. Trn ph khi

lng ESI-MS xut hin pc ion phn t ti m/z 793,6 [M-H]+ . Cng vi cc ph
NMR cho php xc nh cng thc phn t ca HM2 l C41H78O12S.
6'

CH2SO3H
5'
4'

OH

O
1'

OH
2'
3'

OH

CH2
2

HC

1'"

CH2 O

16'"

(CH2)14CH3

O 1"
16"
C (CH2)14CH3
O

Hnh: V.2.4.2.a. Cu trc ca hp cht HM2

Ph cng hng t ht nhn proton (1H-NMR) ca HM2 c trng cho
mt hp cht glycolipit vi s xut hin tn hiu c trng ca cc nhm oximetin
v oximetilen ti 3,1-4,5. S xut hin cc tn hiu ca 2 nhm metyl ti 0,89
(6H, t, J = 6,5Hz), cc tn hiu proton ca mch cc bon no di ti 1,30 (46H,
br s), cc tn hiu proton ca hai nhm metilen gn vi nhm cacbonyl ti 2,33
(4H, m) v 2 nhm -metinlen ti 1,61 (4H, m) khng nh s c mt ca 2
nhnh axit bo no.

Hnh V.2.4.2.b. Ph 1H-NMR ca HM2

S khc bit ln v c trng v chuyn dch ha hc ca ca cc
proton metilen ti v tr C-1 v 3 phn nh v tr th ca n v ng v axit bo
tng ng ti hai v tr ny. Hng s tng tc v s tch vch ca cc proton ca
n v ng tng t nh ng glucose nhng chuyn dch ha hc mt
419

s v tr c s thay i. C th l, proton metilen C-6 nhn c 3,14 v 3,33

khng nh s c mt ca nhm sulphonyl v tr C-6 v gc ng l
sulphoquinovose [259]. S xut hin tn hiu cacbon ti 53,59 trn ph 13CNMR ca HM2 mt ln na khng nh kt lun ny.

Hnh V.2.4.2.c. Ph 13C-NMR ca HM2

Bng V.2.4.2: Cc kt qu ph NMR ca HM2
C

C [260]

Ca, b

63,0 t

63,45 t

70,1 d

70,84 d

67,3 t

66,68 t

1
2
3
4
5

98,9 d
71,4 d
72,7 d
73,4 d
67,8 d

99,28 d
72,22 d
73,97 d
73,58 d
68,77 d

53,6 d

53,59 d

1
2
3
413
14

174,2 s
34,4 t
25,0 t
29,229,8 t
31,9 t

174,46* s
34,66# t
25,22$ t
29,45 29,94 t
32,17 t

Ha,c (J, Hz)

4,20 (dd, 7,0, 12,0)
4,43 (dd, 3,0, 12,0)
5,31 (m)
3,62 (dd, 6,5, 11,0)
3,98 (dd, 5,0, 11,0)
4,83 (d, 3,5)
3,51 (dd, 3,5, 9,5)
3,65 (dd, 9,0, 9,5)
3,38 (dd, 9,0, 9,5)
4,02(m)
3,14 (dd, 6,0, 14,5)
3,33 (dd, 4,0, 14,5)
2,33 (m)
1,61 (m)
1,30 (br s)
1,30 (br s)

420

HMBC (H - C)
C-2, 3, 1
C-2, 1, 1
C-3, 3, 5
C-1
C3, 6
C-1
C5
C-1, 3
C-1, 2

15
16
1
2
3
413
14
15
16
a

22,7 t
14,1 q
174,2 s
34,4 t
25,0 t
29,229,8 t
31,9 t
22,7 t
14,1 q

22,87 t
14,11 q
174,27* s
34,46# t
25,18$ t
29,45 29,94 t
32,17 t
22,87 t
14,11 q
b

1,30 (br s)
0,89 (t, 6,5)
2,33 (m)
1,61 (m)

C-14, 15

1,30 (br s)
1,30 (br s)
1,30 (br s)
0,89 (t, 6,5)

o trong CDCl3+MeOD, 125 MHz, 500 MHz,

C-14, 15

*, #, $

cc gi tr c cng k hiu c th i ch cho nhau.

Trn ph 13C-NMR ca HM2 xut hin cc tn hiu c trng ca 1

cacbon anomer ti 99,28, cc tn hiu ca glyceryl b th hon ton ti 63,45,
70,84 v 66,68. S xut hin ca hai nhm metyl ti 14,11 (nhn 2), hai nhm
cacbonyl ti 174,27 v 174,46, hai nhm -metinlen ti 22,87 (nhn 2) hai
nhm metilen gn vi nhm cacbonyl ti 34,46 v 34,66 mt ln na khng
nh s tn ti ca 2 nhnh axit bo.

Hnh V.2.4.2.d. Ph 13C-NMR v cc ph DEPT ca HM2

xc nh chuyn dch ha hc tng ng gia cacbon v proton, ph
cng hng t ht nhn hai chiu HSQC c o. Cc gi tr chi tit c ch
ra trn bng V.2.4.2.

421

Hnh V.2.4.2.e. Ph HSQC ca HM2

422

Hnh V.2.4.2.f. Ph HMBC ca HM2

xc nh v tr cc nhm th cng nh cu trc tng th ca ton phn
t, chng ti tin hnh o ph cng hng t ht nhn 2 chiu HMBC. Trn
ph ny, s xut hin pc tng tc gia proton anomer H-1 vi C-3 v proton H3 vi cacbon anomer C-1 khng nh v tr th ca gc ng ti C-V.2.4.
Phn tch chi tit cc tng tc trn ph HMBC (hnh V.2.4.2.g v bng V.2.4.2)
cho php d on cng thc ha hc ca HM2 nh a ra trn hnh V.2.4.2.a.
6'

CH2SO3H
5'
4'

OH

O
1'

OH
2'
3'

OH

CH2
2

HC

CH2 O

1'"

O1"
C

16'"

CH2CH2(CH2)12CH3
16"

CH2CH2(CH2)12CH3

Hnh V.2.4.2.g. Mt s tng tc HMBC chnh ca HM2

Cc kt qu phn tch trn c mang so snh vi cc d kin ph ca hp
cht (2S)1,2-di-O-palmitoyl-3-O-(6-sulfo--D-quinovopyranosyl) glycerol [260].
S trng khp hon ton v cc gi tr ph tng ng cho php khng nh hp
cht HM2 l (2S)1,2-di-O-Palmitoyl-3-O-(6-sulfo--D-quinovopyranosyl)
Glycerol.

423

Hnh V.2.4.2.h. Ph ESI-MS ca HM2

Hp cht ny c phn lp t cc loi to nu Ishige okamurai v
Sargassum parivesiculosum. Tuy nhin y l ln u tin hp cht ny c
phn lp t chi Gellius. Theo cc ti liu cng b [260], hp cht ny th hin
hot tnh khng Pyricularia oryzae, hot tnh gy c t bo i vi cc dng t
bo u P388 v HL60. Ngoi ra n cn th hin hot tnh gy c t bo yu i
vi cc dng t bo u ngi l MCF7 v A549. Hp cht ny c phn lp vi
hm lng kh cao t loi hi min G. varius. iu ny gi m cho nhng nghin
cu su hn v hp cht ny theo hng ng dng lm dc liu. Mt iu th v
na l, gn y M c mt s bng c quyn sng ch c cp v kh
nng ng dng lm thuc ca cc dn xut thuc lp cht sulfono-glycolipid
[261]. Theo cc tc gi ny, lp cht ny thng th hin hot tnh gy c t bo
v l cc cht c ch enzym tng hp DNA-polymerase.
V.2.4.3. Hp cht HM3: Mui natri ca (2S)1-O-Hexadecanoyl-3-O-(6-sulfo-D-quinovopyranosyl) Glycerol
Hp cht HM3 cng nhn c di dng bt v nh hnh mu trng.
Cng thc phn t ca HM3 c xc nh l C25H47NaO11S bng cc d kin
ph khi (m/z 555,3 [M-Na]+, 6.1,2 [M+Na]+) v ph cng hng t ht nhn.

424

Hnh V.2.4.3.a. Ph 1H-NMR ca HM3

6'

CH2SO3Na
5'
4'

OH

O
1'

OH
2'
3'

OH

CH2
2

HC

OH

CH2 O

1"

16"

(CH2)14CH3

Hnh V.2.4.3.b. Cu trc ha hc ca HM3

Ph cng hng t ht nhn proton (1H-NMR) cng c trng cho mt
hp cht glycolipit vi s xut hin cc tn hiu c trng ca cc nhm oximetin
v oximetilen ti 2,9-4,2. S xut hin cc tn hiu ca 1 nhm metyl ti 0,93
(3H, t, J = 6,5Hz), cc tn hiu proton ca mch cc bon no di ti 1,30 (26H,
br s), cc tn hiu proton ca 1 nhm metilen gn vi nhm cacbonyl ti 2,39
(2H, m) v 1 nhm -metinlen ti 1,63 (2H, m) khng nh s c mt ca 1
nhnh axit bo no.

425

Hnh V.2.4.3.c. Ph 13C-NMR ca HM3

S khc bit ln v c trng v chuyn dch ha hc ca ca cc
proton metilen ti v tr C-1 v 3 phn nh v tr th ca n v ng v axit bo
tng ng ti hai v tr ny. Hng s tng tc v s tch vch ca cc proton ca
n v ng tng t nh ng glucose nhng chuyn dch ha hc mt
s v tr c s thay i. C th l, proton metilen C-6 nhn c 2,95 v 3,38
khng nh s c mt ca nhm sulphonyl v tr C-6 v gc ng l
sulphoquinovose [259]. S xut hin tn hiu cacbon ti 54,19 trn ph 13CNMR ca HM3 mt ln na khng nh kt lun ny.
6'

CH2SO3Na
5'
4'

O
1'

OH

OH

2'
3'

OH

CH2
2

HC

OH

CH2 O

1"

16"

(CH2)14CH3

Hnh V.2.4.3.d. Mt s tng tc HMBC chnh ca HM3

Bng V.2.4.3. Kt qu ph cng hng t ht nhn ca HM3
C

C [262]

Ca, b

66,4 t

66,53 t

69,2 d

69,79 d

70,0 t

70,49 t

Ha, c (J, Hz)

4,12 *
4,22 (dd, 6,5, 13,5)
4,11 (m)
4,08 *
3,42 *

426

HMBC (H to C)
C-2, 3, 1

C-1, 2, 1

1
2
3
4
5

99,6 d
72,5 d
73,9 d
73,5 d
69,2 d

100,12 d
73,59 d
75,07 d
74,88 d
69,79 d

53,3 d

54,19 d

1
2
3
413
14
15
16

176,1 s
34,9 t
25,7 t

175,59 s
34,93 t
25,97 t
30,22 30,76 t
33,04 t
23,70 t
14,42 q

29,2-29,8 t
32,9 t
23,6 t
14,8 q

4,81 (d, 3,5)

3,43 *
3,68 (t, 9,0)
3,12 (t, 9,0)
4,12 *
2,95 (dd, 9,5, 14,5)
3,38 *
2,39 (m)
1,63 (m)

C-3, 3, 5
C-2, 4, 5
C-3, 5, 6
C-1, 3, 4, 6
C-4, 5
C-1, 3
C-1, 2

1,30 (br s)
1,30 (br s)
1,30 (br s)
0,93 (t, 6,5)

C-14, 15

o trong MeOD, b 125 MHz, c 500 MHz, * tn hiu b che lp

Trn ph 13C-NMR xut hin tn hiu ca mt nhm metyl ti 14,42,

mt tn hiu ca nhm cacbonyl ti 175,59; mt nhm -metinlen ti 23,70;
mt nhm metilen gn vi nhm cacbonyl ti 34,93 mt ln na khng nh s
tn ti ca mt nhnh axit bo duy nht. Cc tn hiu ca cc nhm metilen xut
hin ti 30,22 - 30,76. Tn hiu cc bon anomer xut hin ti 100,12; cc tn
hiu cacbon ca cc n v ng xut hin ti 54,19 - 75,07; cc tn hiu ca
glyceryl b th 2 v tr ti 66,53, 69,79 v 70,49.

Hnh V.2.4.3.e. Ph 13C-NMR v cc ph DEPT ca HM3

427

Hnh V.2.4.3.f. Ph HSQC ca HM3

xc nh chuyn dch ha hc tng ng gia cacbon v proton, ph
cng hng t ht nhn hai chiu HSQC c o. Cc gi tr chi tit c ch
ra trn bng V.2.4.3.

Hnh V.2.4.3.g. Ph HMBC ca HM3

428

 xc nh v tr cc nhm th cng nh cu trc tng th ca ton phn

t, chng ti tin hnh o ph cng hng t ht nhn 2 chiu HMBC. Trn
ph ny, s xut hin pc tng tc gia proton anomer H-1 vi C-3 v proton H3 vi cacbon anomer C-1 khng nh v tr th ca gc ng ti C-3. Phn
tch chi tit cc tng tc trn ph HMBC (hnh V.2.4.3.d v bng V.2.4.3) cho
php d on cng thc ha hc ca HM3 nh a ra trn hnh V.2.4.3.b.

Hnh V.2.4.3.h. Ph ESI-MS ca HM3

Cc kt qu phn tch nu trn c mang so snh vi cc d kin ph
c cng b ca hp cht mui natri ca (2S)1-O-Hexadecanoyl-3-O-(6-sulfo-D-quinovopyranosyl) Glycerol [262]. S trng khp hon ton v cc gi tr
ph tng ng cho php khng nh HM3 l mui natri ca (2S)1-OHexadecanoyl-3-O-(6-sulfo--D-quinovopyranosyl) Glycerol. Hp cht ny
c phn lp t loi cu gai Anthocidaris crassispina di dng hn hp vi
ng phn cha axit bo l axit myristic vi t l 96/4. Tuy nhin y l ln u
tin n c phn lp di dng n cht t cc loi hi min thuc chi Gellius.
V.2.4.4. Hp cht HM4: Cholesterol
Hp cht HM4 nhn c di dng tinh th hnh kim c mu trng. Th
nghim trn sc k lp mng (TLC) vi thuc th c hiu (phun dung dch
H2SO4 10% ri h nng t t cho mu tm hng, sau chuyn dn sang tm
xanh) cho thy y l mt steroit. Hp cht ny thu c vi hm lng rt cao
cng vi gi tr rf = 0,35 trn TLC vi h dung mi l Hexan/Axeton 7/1 v gi
tr nhit nng chy (mp 148,5oC) ca n gi ti hp cht cholesterol, mt
sterol ph bin nht trong gii ng vt. Cng vi s ph hp hon ton v s liu
429

ph 1H- v 13C-NMR (xem phn 2.5.5) ca HM4 so vi ti liu cng b [220]

cho php khng nh hp cht ny chnh l cholesterol.
21

26
20

18

19

11

13
14

HO

17

25

27

15

10
3

22

7
6

Hnh V.2.4.4. Cu trc ha hc ca HM4

V.2.4.5. Hp cht HM5: 5,8-Epidioxycholest-6-en-3-ol
Hp cht HM5 nhn c di dng cht tinh th hnh kim mu trng.
Ph 1H-NMR ca HM5 c trng cho mt hp cht steroit, trong xut hin
cc tn hiu ca 5 nhm metyl ti 0,86 (3H, s), 0,90 (3H, d, J = 6,5Hz), 0,91
(3H, d, J = 6,5Hz), 0,92 (3H, s), 0,96 (3H, d, J = 6,5Hz); cc proton olefin ti
6,30 (1H, d, J = 8,5Hz) v 6,55 (1H, d, J = 8,5Hz); mt proton gn vi cacbon c
ni vi nguyn t xy ti 3,80 (1H, m).

Hnh V.2.4.5.a. Ph 1H-NMR ca HM5

Ph 13C-NMR xut hin cc tn hiu ca mt steroit 27 cacbon gm c 4

cacbon bc 4, 8 metin, 10 metilen, v 5 metyl. Mt ni i b th 2 ln c xc
nh ti 131,75 (d)/136,80 (d); 2 cacbon bc 4 ni vi nguyn t oxy ti 80,87
(s) v 83,49 (s), 1 cacbon metin gn vi nguyn t xy ti 66,98 (d).

430

Bng V.2.4.5. Cc s liu ph NMR ca HM5

C

C [251]

C a, b

39,5 t

40,65 t

1,96/2,02

30,1 t

30,87 t

1,54/1,81 (m)

66,5 d

66,98 d

3,80 (m)

36,8 t

37,17 t

1,95/2,15

82,1 s

83,49 s

135,4 d

136,80 d

6,30 (d, 8,5)

130,8 d

131,75 d

6,55 (d, 8,5)

79,5 s

80,78 s

51,1 d

52,74 d

1,52

10

36,9 s

37,78 s

11

20,6 t

21,58 t

1,06 (m)/1,59

12

39,5 t

40,82 t

1,22

13

44,7 s

45,95 s

14

51,6 d

52,74 d

1,58 (d, 3,5)

15

23,4 t

24,41 t

1,21/1,52

16

28,2 t

29,25 t

1,45

17

56,4 d

57,81 d

1,20

18

12,6 q

13,04 q

0,86 (s)

19

18,6 q

19,07 q

0,92 (s)

20

35,2 d

36,52 d

1,60

21

18,1 q

18,59 q

0,96 (d, 6,5)

22

35,9 t

37,16 t

1,73 (m)

23

23,8 t

24,90 t

1,32 (m)

24

39,5 t

40,65 t

1,24

25

28,0 d

29,25 d

1,45

26

22,5 q

22,92 q

0,90 (d, 6,5)

23,17 q

0,91 (d, 6,5)

22,8 q

27
a

o trong CD3OD, 125MHz, 500MHz

431

Ha, c (J, Hz)

Hnh V.2.4.5.b. Ph 13C-NMR ca HM5

T cc phn tch nu trn c th s b xc nh c HM5 c b khung
cacbon tng t nh ca HM4 tuy nhin c s chuyn dch v tr ca lin kt
i v s xut hin 2 cacbon bc bn gn vi nguyn t xy. chuyn dch ha
hc tng i cao ti cc v tr cacbon bc bn lin kt vi nguyn t xy gi
ti s c mt ca mt cu peroxit.

Hnh V.2.4.5.c. Ph 13C-NMR v cc ph DEPT ca HM5

432

22

21
18

17

11

27

9
10

HO

26

25

13

19
1

20

15

H
7 O

Hnh V.2.4.5.d. Cu trc ha hc ca HM5

Cui cng, cu trc ha hc ca HM5 c xc nh l 5,8Epidioxycholest-6-en-3-ol bi s so snh v s trng khp hon ton (bng
V.2.4.5) cc gi tr tng ng v s liu ph 13C-NMR v cc c tnh vt l,
ha hc ca HM5 vi hp cht ny c cng b [251]. Hp cht ny cng
c chng ti phn lp t loi cu gai Diadema setosum v cc kt qu nghin
cu v hot tnh sinh hc in vitro cho thy hp cht ny th hin hot tnh gy
c t bo cao i vi nhiu dng t bo ung th nh dng KB (t bo ung th
biu m, IC50 = 2,0g/ml), FL (t bo ung th mng t cung, IC50 = 3,9g/ml) v
Hep-2 (t bo ung th gan, IC50 = 2,4g/ml) [251].
V.2.4.6. Hp cht HM6: Cholest-7-ene-3,5,6-triol
Hp cht HM6 nhn c di dng tinh th hnh kim mu trng. Cng
thc phn t ca HM6 c xc nh l C27H46O3 bng cc d kin ph khi (m/z
401,2 [M-H2O+H]+, 383,2 [M-2H2O+H]+, 365,2 [M-3H2O+H]+) v ph cng
hng t ht nhn.

Hnh V.2.4.6.a. Ph 1H-NMR ca HM6

433

21
18
11

HO

10
5
4

27

13
9

2
3

26
25

17

19
1

22
20

15

OH
OH

Hnh V.2.4.6.b. Cu trc ha hc ca HM6

Tng t cc hp cht HM4 v HM5, ph cng hng t ht nhn 1HNMR ca HM6 c trng cho mt hp cht steroit. Trong xut hin cc tn
hiu c chng ca 5 nhm metyl ti 0,60 (3H, s, H-18), 0,88 (3H, d, J =
6,5Hz, H-26), 0,89 (3H, d, J = 6,5Hz, H-27), 0,94 (3H, d, J = 6,5Hz, H-21) v
1,09 (3H, s, H-19); mt proton olefin ti 5,37 (1H, m, H-7) v 2 proton gn vi
cacbon c ni vi nguyn t oxy ti 4,09 (1H, m, H-3) v 3,63 (1H, br s, H-6).

Hnh V.2.4.6.c. Ph 13C-NMR ca HM6

Ph 13C-NMR ca HM6 xut hin cc tn hiu c trng ca mt hp cht
steroit 27 cabon. Trong , mt ni i b th 3 ln c xc nh ti 117,56 (d,
C-7)/144,05 (s, C-8); nm nhm metyl ti 12,11 (C-18), 18,84 (C-19 v C-21),
22,56 (C-26) v 22,82 (C-27); ba cacbon ni trc tip vi nguyn t xy ti
67,74 (d, C-3), 76,01 (s, C-5) v 73,69 (d, C-6) (iu ny cng c khng nh
bi s xut hin pc ion phn mnh ti m/z 365,2 do b loi mt 3 phn t nc
ca HM6).

434

Hnh V.2.4.6.d. Ph 13C-NMR v cc ph DEPT ca HM6

xc nh gi tr ph cng hng t ht nhn proton gn vi cc bon
tng ng, ph cng hng t ht nhn 2 chiu tng tc trc tip HSQC c
tin hnh. Kt qu c ch ra bng V.2.4.6.
Bng V.2.4.6. Kt qu ph NMR ca HM6.
C

C [263]

C a, b

Ha, c (J, Hz)

1
2
3
4

31,8 t
34,4 t
68,4 d
40,9 t

32,98 t
30,87 t
67,74 d
39,49 t

5
6
7
8
9
10
11
12
13
14
15
16
17
18

77,0 s
74,4 d
119,2 d
143,8 s
44,5 d
38,2 s
23,0 t
40,6 t
44,8 s
55,9 d
24,0 t
28,7 t
57,6 d
12,4 q

76,01 s
73,69 d
117,56 d
144,05 s
43,49 d
37,15 s
22,08 t
39,49 t
43,89 s
54,72 d
22,94 t
27,77 t
56,25 d
12,10 q

1,45/1,85
1,55/1,62
4,09 (m)
1,79 (dd, 5,0, 13,0)
2,10 (dd, 13,0, 13,0)
3,63 (br s)
5,37 (m)
1,96
1,59
1,32/1,80
1,91
1,35/1,60
1,32/1,92
1,26
0,60 (s)

435

HMBC (H
C)

C-2, 3, 5, 10
C-5, 7, 8
C-5

C-12, 13, 14, 17

19
20
21
22
23
24
25
26
27

18,8 q
37,2 d
19,3 q
36,0 t
-

18,84 q
36,14 d
18,84 q
36,07 t
23,93 t
39,37 t
28,02 d
22,56 q
22,82 q

1,09 (s)
1,39
0,94 (d, 6,5)
1,11/1,39
1,55
1,53
1,52
0,88 (d, 6,5)
0,89 (d, 6,5)

C-1, 9, 10
C-17, 20, 22

C-24, 25, 27
C-24, 25, 26

o trong CDCl3, b125MHz, c 500MHz

Dng pc a vch ti 4,09 c trng cho proton 3-carbinol ca mt A/B

trans-streroit. S chuyn dch v pha trng thp ca tn hiu c trng cho cc
cht 3-hydroxysterol mang mt nhm 5-hydroxyl. Cc d kin ny cng vi s
ph hp ca tn hiu cng hng ca nhm metyl (C-18) ti 0,60 (s) vi gi tr
tng ng vi mt 7-sterol gi cho cu trc 7-ene-3,5,6-triol. S gim
chuyn dch ha hc ca tn hiu metyl C-19 ti 1,09 (s) l du hiu cho cu hnh
ca nhm hydroxyl v tr C-6 [264].

Hnh V.2.4.6.e. Ph HSQC ca HM6

436

Hnh V.2.4.6.f. Ph HMBC ca HM6

Hnh V.2.4.6.g. Ph khi lng ESI-MS ca HM6

437

21
18

10

27

13
9

HO

26
25

17

11
19
1

22
20

15

7
6

4
OH

OH

Hnh V.2.4.6.h. Mt s tng tc HMBC chnh ca HM6

S ph hp hon ton v cc s liu ph 1H-NMR v 13C-NMR ti cc v
tr nhnh bn tng ng ca HM6 so vi HM5 cho php khng nh hp cht ny
c cng nhnh bn vi HM5. Cu trc ha hc ca HM6 c xc nh nh ch
ra hnh V.2.4.6.b bng s phn tch chi tit cc tng tc trn ph HMBC cng
vi ph hp v s liu ph 13C-NMR vi cc hp cht c cng khung nhng khc
i cht nhnh bn (bng V.2.4.6) [263]. T cc phn tch nu trn, hp cht
HM6 c xc nh l cholest-7-ene-3,5,6-triol. Hp cht ny c phn
lp trc t cc loi Patinopecten yessoensis v Aplysia juliana. Tuy nhin,
y l ln u tin n c phn lp t chi Gellius.
V.2.4.7. Hp cht HM7: 6-Methoxycholest-7-ene-3,5-diol
Hp cht HM7 cng nhn c di dng tinh th hnh kim mu trng.
Cng thc phn t ca n c xc nh l C28H48O3 t cc d kin ph khi ESI-MS
(m/z 415.3 [M-H2O+H]+, 397.2 [M-2H2O+H]+, 38V.2.4.3 [M-2H2O-CH2+H]+,
365.3 [M-2H2O-CH2-H2O+H]+, 431.1 [M-H]+) v ph cng hng t ht nhn.

Hnh V.2.4.7.a. Ph 1H-NMR ca HM7

438

21
18
11

HO

27

13
8

10
5
4

25

2
3

26

17

19
1

22
20

15

OH
OCH3

Hnh V.2.4.7.b. Cu trc ha hc ca HM7

Ph 1H-NMR ca HM7 gn nh trng khp hon ton vi hp cht HM6
ngoi tr s xut hin thm tn hiu ca mt nhm metoxy ti 3,40 (3H, s).
iu ny cho thy HM7 l mt dn xut metyl te ca HM6.
Bng V.2.4.7. Kt qu ph NMR ca HM7
C

C ca
HM6

C [264]

C a, b

H a, c (J, Hz)

32,98 t

32,8 t

32,78 t

1,50/1,90

30,87 t

30,9 t

30,92 t

1,60/1,68

67,74 d

67,8 d

67,86 d

4,05 (m)

39,49 t

39,5 t

39,51 t

1,85 (dd, 5,0, 13,0)

2.15 (dd, 13,0, 13,0)

76,01 s

76,3 s

76,36 s

73,69 d

82,5 d

82,48 d

3,17 (br d, 5,0)

117,56 d

115,0 d

114,97 d

5,41 (m)

144,05 s

143,7 s

143,75 s

43,49 d

43,9 d

43,92 d

2,05

10

37,15 s

37,3 s

37,27 s

11

22,08 t

22,2 t

22,19 t

1,59

12

39,49 t

39,5 t

39,51 t

1,34/1,90

13

43,89 s

44,0 s

43,99 s

14

54,72 d

54,9 d

54,93 d

1,91

15

22,94 t

22,9 t

22,94 t

1,39/1,65

16

27,77 t

27,8 t

27,81 t

1,32/1,92

17

56,25 d

56,2 d

56,25 d

1,26

439

18

12,10 q

12,1 q

12,07 q

0,60 (s)

19

18,84 q

18,4 q

18,36 q

1,00 (s)

20

36,07 d

36,2 d

36,16 d

1,42

21

18,84 q

18,8 q

18,84 q

0,93 (d, 6,5)

22

36,14 t

36,1 t

36,10 t

1,22/1,43

23

23,93 t

23,9 t

23,93 t

1,57

24

39,37 t

39,6 t

39,62 t

1,53

25

28,02 d

28,0 d

28,02 d

1,52

26

22,56 q

22,6 q

22,56 q

0,88 (d, 6,5)

27

22,82 q

22,8 q

22,82 q

0,87 (d, 6,5)

58,2 q

58,27 q

3,40 (s)

OCH3
a

o trong CDCl3, 125MHz, 500MHz

Hnh V.2.4.7.c. Ph 13C-NMR ca HM7

Ph 13C-NMR ca HM7 c trng cho mt steroit 28 cacbon. S tn ti
mt nhm metoxy cng c xc nh ti 58,27 (q). Hu ht cc gi tr ph u
trng khp so vi hp cht HM6, ngoi tr s thay i cc v tr C-5 n C-10
(bng V.2.4.7). c bit ti v tr C-6 v C-7 c s thay i rt mnh t
73,69 ln 82,48 v t 117,56 v 114,97. iu ny cho php khng nh nhm
metoxy phi nm v tr C-6.

440

Hnh V.2.4.7.d. ph 13C-NMR v cc ph DEPTca HM7

T tt c cc phn tch trn v s ph hp hon ton v s liu ph so vi
s liu ca 6-methoxycholest-7-ene-3,5-diol [264], hp cht HM7 c xc
nh l 6-methoxycholest-7-ene-3,5-diol (hnh V.2.4.7.b). Hp cht ny
c phn lp trc t loi hi min Aplysia juliana. Tuy nhin y l ln u
tin n c phn lp t chi Gellius.

Hnh V.2.4.7.e. Ph khi lng ESI-MS ca HM7

441

V.2.4.8. Hp cht HM8: Metyl este ca axit (5Z,8Z,11Z,14Z,17Z)eicosapentaenoic

Hp cht HM8 nhn c di dng cht du khng mu. Ph 1H-NMR
xut hin cc tn hiu c trng ca 10 proton olefin ti 5,38 (10H, m); 1 nhm
metoxy ti 3,68 (3H, s); 1 nhm metyl ti 0,98 (3H, t, J = 7,5Hz) v cc tn
hiu ca cc nhm metilen ti 1,71 (2H, m), 2,10 (4H, m), 2,32 (2H, t, J =
7,5Hz) v 2,83 (8H, m).
O
11

3
4

14

17

1
2

20

Hnh V.2.4.8.a. Cu trc ha hc ca HM8

Hnh V.2.4.8.b. Ph 1H-NMR ca HM8

Ph 13C-NMR xut hin cc tn hiu c trng ca 1 nhm cabonyl ti
174,06 (s); 5 ni i b th 2 v tr ti 132,06 (d), 128,96 (d), 128,87 (d), 128,60
(d), 128,28 (d), 128,25 (d), 128,15 (d), 128,13 (d), 127,90 (d) v 127,04 (d); 1
nhm metyl ti 14,27 (q); 1 nhm metoxy ti 51,48 v cc tn hiu ca 7
nhm metylen. Gi tr chuyn dch ha hc tng i thp ca mt nhm
metoxy chng t n l mt nhm metyl este.

442

Hnh V.2.4.8.c. Ph 13C-NMR ca HM8

Bng V.2.4.8. Kt qu ph NMR ca HM8

C
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
OCH3

C [265]
173,60
33,72
25,72
24,82
129,00
128,22
25,51
128,73
128,22
25,59
128,53
128,07
25,59
128,07
126,98
25,59
127,84
131,70
20,02
14,25

C a, b
174,06 s
33,47 t
26,58 t
24,81* t
128,97# d
127,90# d
25,65 t*
128,87# d
128,59# d
25,65* t
128,28# d
128,25# d
25,65* t
128,15# d
128,13# d
25,56* t
127,04# d
132,06 d
20,57 t
14,27 q
51,48 q

o trong CDCl3, b125MHz, c 500MHz, *, # gi tr c th i v tr cho nhau

443

Ha, c (J, Hz)

2,32 (t, 7,5)
1,71 (m)
2,10 (m)
5,38 (m)
5,38 (m)
2,83
5,38 (m)
5,38 (m)
2,83
5,38 (m)
5,38 (m)
2,83
5,38 (m)
5,38 (m)
2,83
5,38 (m)
5,38 (m)
2,10 (m)
0,98 (t, 7,5)
3,67 (s)

Hnh V.2.4.8.d. Ph 13C-NMR v cc ph DEPT ca HM8

Vi cc d kin nu, chng ta c th s b xc nh HM8 l dng metyl este
ca mt axit mch di 20 cacbon trong c 5 ni i ni mch.

Hnh V.2.4.8.e. Ph khi lng ESI-MS ca HM8

444

T tt c cc phn tch trn, s liu ph NMR ca HM8 c so snh vi

s liu c cng b ca hp cht metyl este ca axit (5Z,8Z,11Z,14Z,17Z)eicosapentaenoic v cho thy s ph hp hon ton cc gi tr tng ng [265].
Cng vi kt qu ph khi lng ESI-MS ti m/z 317.1 [M+H]+, 315.0 [M-H]+
tng ng vi cng thc phn t C21H32O2 cho php khng nh cu trc ha hc
ca hp cht ny l metyl este ca axit (5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic.
V.2.4.9. Hp cht HM9: Phytol
Hp cht HM9 nhn c di dng cht du khng mu. Ph 1H-NMR
ca n c trng cho mt hp cht tecpen. Trong c cc tn hiu proton c
trng ca 5 nhm metyl ti 0,86 (12H) v 1,67 (3H, s, H-20); mt nhm
oximetilen ti 4,14 (2H, d, J = 7,0Hz, H-1), mt ni i b th 3 v tr ti 5.40
(1H, t, J = 7,0Hz, H-2).

Hnh V.2.4.9.a. Ph 1H-NMR ca HM9

Ph 13C-NMR cho thy HM9 l mt hp cht ditecpen trong c 5 nhm
metyl, 1 cacbon bc 4, bn cacbon bc 3 v 10 cacbon bc 1. Trong , 5 nhm
metyl c xc nh ti 16,18, 19,77, 19,73, 22,73, v 22,64; mt nhm
oximetilen ti 59,40; mt ni i b th 3 v tr ti 123,17 (d)/140,20 (s).

445

Hnh V.2.4.9.b. Ph 13C-NMR ca HM9

Bng V.2.4.9. Kt qu ph NMR ca HM9
C

C [266]

59,4 t

C a, b

H a, c (J, Hz)

59,40 t

4,14 (d, 7,0)

123,2 d

123,17 d

5,40 (t, 7,0)

140,2 s

140,20 s

39,9 t

39,90 t

1,99 (t, 7,5)

25,2 t

25,17 t

36,8 t

36,70 t

32,8 d

32,82 d

37,5 t

37,46 t

24,5 t

24,49 t

10

37,4 t

37,32 t

11

32,7 d

32,72 d

12

37,4 t

37,39 t

13

24,9 t

24,82 t

14

39,4 t

39,40 t

15

28,0 d

27,99 d

16

22,7 q

22,64 q

17

22,8 q

22,73 q

446

1,02-1,56 (19H)

0,86 (12H)

18#

19,8 q

19,73 q

19#

19,8 q

19,77 q

20

16,2 q

16,18 q

1,67 (s)

o trong CDCl3, b125MHz, c 500MHz, *, #gi tr c th i v tr cho nhau

Hnh V.2.4.9.c. Ph 13C-NMR v cc ph DEPT ca HM9

17

15

18

19

11

20
3

OH

16

Hnh V.2.4.9.d. Cu trc ha hc caHM9

Cc s liu ph NMR ca HM9 gi ti hp cht trans-phytol, mt

ditecpen rt ph bin trong thin nhin. T , s liu ph 13C-NMR ca hp
cht ny c mang so snh vi s liu ca trans-phytol c cng b [266].
S trng khp hon ton v gi tr ph 1H-NMR, 13C-NMR v DEPT ti cc v tr
tng ng cho php khng nh cu trc ha hc ca HM9 l trans-phytol.
V.2.4.10. Hp cht HM10: Glycerol 1-(5Z,8Z,11Z,14Z-eicosatetraenoate)
Hp cht HM10 nhn c di dng cht du khng mu. Ph 1H-NMR
xut hin cc tn hiu c trng ca 8 proton olefin ti 5,37 (m); cc tn hiu
ca cc proton ni vi cabon c gn vi nguyn t xy ti 3,72 (1H, dd, J =
447

4,0, 11,5Hz, H-1a), 3,62 (1H, dd, J = 5,5, 11,5Hz, H-1b), 3,95 (1H, m, H-2), 4,23
(1H, dd, J = 4,5, 11,5Hz, H-3a) v 4,17 (1H, dd, J = 6,5, 11,5Hz, H-3b); mt
nhm metyl ti 0,91 (3H, t, J = 7,0Hz, H-20) v cc tn hiu ca cc nhm
metilen.

Hnh V.2.4.10.a. Ph 1H-NMR ca HM10

3

HO

OH
2

O
5'

8'

1'

20'
11'

14'

Hnh V.2.4.10.b. Cu trc ha hc ca HM10

Ph 13C-NMR xut hin cc tn hiu c trng ca mt nhm cacbonyl ti
174,06 (s), 4 ni i b th 2 v tr ti 130,54 (d), 129,07 (d), 128,76 (d),
128,65 (d), 128,32 (d), 128,11 (d), 127,86 (d), 127,55 (d); cc tn hiu ca mt
glyxerin b th mt v tr v khng i xng ti 63,36 (t, C-1), 70,28 (d, C-2) v
65,27 (t, C-3) [267]; 1 nhm metyl ti 14,07 (q, C-20) v cc tn hiu ca cc
nhm metilen. Nh vy c th s b xc nh HM10 l dng este ca glyxerin
vi mt axit bo c 4 lin kt i.

448

Bng V.2.4.10. Kt qu ph NMR ca HM10

C

C [268]

C a, b

75,1 t

63,36 t

H a, c (J, Hz)
3,72 (dd, 4,0, 11,5)
3,62 (dd, 5,5, 11,5)

62,5 d

70,28 d

3,95 (m)

75,1 t

65,27 t

4,23 (dd, 4,5, 11,5)

4,17 (dd, 6,5, 11,5)

173,8 s

174,06 s

33,7 t

33,51 t

2,39 (t, 7,5)

29,3 t

29,34 t

1,74 (q, 7,5)

2,85 (m) $

24,8 t

24,74 t

129,0 d

129,07 d #

5,37 (m)

127,8 d

127,86 d #

5,37 (m)

2,85 (m) $

26,5 t

26,53 t

128,8 d

128,76 d #

5,37 (m)

128,6 d

128,65 d #

5,37 (m)

2,85 (m) $

10

25,64 t

25,65 t

11

128,3 d

128,32 d #

5,37 (m)

12

128,1 d

128,11 d #

5,37 (m)

2,15 (m) $

13

25,62 t

25,64 t

14

127,5 d

127,55 d #

5,37 (m)

15

130,5 d

130,54 d

5,37 (m)

16

27,2 t

27,24 t

2,08 (m) $

17

29,3 t

29,34 t

18

31,5 t

31,53 t

19

22,6 t

22,58 t

20
a

14,1 q
b

o trong CDCl3, 125MHz, 500MHz,

14,07 q
*, #,$

gi tr c th i v tr cho nhau

449

1,33 (6H, m)
0,91 (t, 7,0)

Hnh V.2.4.10.c. Ph 13C-NMR ca HM10

Hnh V.2.4.10.c. Ph 13C-NMR v cc ph DEPTca HM10

T cc phn tch nu trn, cc s liu NMR ca HM10 c mang so snh
vi s liu c cng b ca Glycerol 2-(5Z,8Z,11Z,14Z-eicosatetraenoate)
[268] v thy s ph hp hon ton ti cc v tr tng ng ca nhnh axit. Ngoi
tr s khc nhau cc v tr C-1, 2 v 3 do tnh cht i xng ca hp cht ny.
450

Cng vi kt qu ph khi lng ESI-MS ti m/z 379.2 [M+H]+, 361.2 [MH2O+H]+, 287.2 [M-Glycerol+H]+, 377.1 [M-H]+ tng ng vi cng thc phn
t C23H38O4 cho php khng nh HM10 l Glycerol 1-(5Z,8Z,11Z,14Zeicosatetraenoate).

Hnh V.2.4.10.e. Ph khi lng ESI-MS ca HM10

V.2.5. nh gi hot tnh sinh hc ca cc hp cht
V.2.5.1. Hot tnh gy c t bo
Trc tin, tin hnh nh gi gi tr CS% ca cc hp cht. Gi tr CS%
hay chnh l gi tr biu hin lng t bo sng st sau khi cho tip xc vi cc
mu chit c tnh theo cc cng thc trnh by trong phn quy trnh. Cc
cht c CS% < 50 c coi l c hot tnh v s c tip tc tin hnh xc nh
gi tr IC50. Kt qu th nghim cho thy: Hp cht HM5 th hin hot tnh trn
c 3 dng t bo th nghim. Hp cht HM1 v HM2 th hin hot tnh 2/3
dng t bo th nghim. Hp cht HM3 th hin hot tnh 1/3 dng t bo th
nghim.
Bng V.2.5.1. Hot tnh gy c t bo ca cc cht
Hp cht
DMSO
Ellipthine
HM1
HM2
HM3
HM4

Gi tr CS% (Cell survival)

KB
1000,0
1,071,07
40,20,5
72,12,6
76,71,2
64,20,9

FL
1000,0
1,520,00
81,20,9
34,22,5
74,22,2
71,11,2

Hep-G2
100,00,0
1,340,63
44,21,5
39,21,1
29,40,7
82,21,6
451

IC50 (g/ml)
KB
0,22
3,51
(-)
(-)
(-)

FL
0,11
(-)
4,06
(-)
(-)

Hep-G2
0,19
3,15
2,89
2,71
(-)

HM5
HM6
HM7
HM8
HM9
HM10

30,11,0
56,13,2
66,71,4
84,21,7
65,72,4
79,64,1

27,31,2
73,22,1
69,81,6
91,12,5
78,13,6
74,92,4

24,21,5
69,22,6
74,21,2
72,20,7
66,71,3
56,71,8

2,00
(-)
(-)
(-)
(-)
(-)

3,93
(-)
(-)
(-)
(-)
(-)

2,40
(-)
(-)
(-)
(-)
(-)

Cc kt qu nghin cu trn cho thy s ph hp vi cc ti liu cng

b. Cc hp cht glycolipit th hin nhiu hot tnh qu bu trong c hot tnh
gy c t bo. C th l hp cht HM2 th hin hot tnh gy c t bo i vi
cc dng t bo u P388 v HL60. Ngoi ra n cn th hin hot tnh gy c t
bo i vi cc dng t bo u ngi l MCF7 v A549 [260]. Gn y M
c mt s bng c quyn sng ch c cp v kh nng ng dng lm thuc
ca cc dn xut thuc lp cht sulfono-glycolipid [261]. Theo cc tc gi ny,
lp cht ny thng th hin hot tnh gy c t bo v l cc cht c ch enzym
tng hp DNA-polymerase. Cc hp cht ny c phn lp vi hm lng kh
cao t loi hi min G. varius sinh sng rt ph bin ti cc vng bin Vit Nam.
iu ny gi m cho nhng nghin cu su hn v hp cht ny theo hng ng
dng lm dc liu.
V.2.5.2. Hot tnh khng vi sinh vt kim nh
Tt c cc hp cht phn lp c t loi hi min Gellius varius c
th nghim v nh gi hot tnh khng vi sinh vt kim nh trn cc dng vi
sinh vt gy bnh tiu biu nht thuc cc nhm vi khun Gr(-); vi khun Gr(+);
nm mc v nm men. nh gi s b hot tnh, chng ti pha mu nng
theo tiu chun sng lc s b v tm cc mu th hin hot tnh khng vi sinh
vt kim nh nng . Cc mu c biu hin dng tnh bc s b ny
c pha long theo cc thang nng thp dn, t 5-10 thang nng tnh
gi tr nng ti thiu (MIC) m vi sinh vt b c ch pht trin gn nh
hon ton.
Bng V.2.5.2. Kt qu xc nh gi tr MIC ca HM1-HM3
Hp cht

Vi khunGr(-)

Nng c ch ti thiu(MIC: g/ml)

Vi khunGr(+)
Nm mc

Nm men

E.coli

P.aeru

B.subti

S.aureus

Asp.niger

F.oxy

S.cere

C.albic

HM1

(-)

150

(-)

200

(-)

(-)

(-)

(-)

HM2

(-)

100

100

(-)

(-)

50

200

(-)

HM3

200

(-)

200

(-)

(-)

(-)

200

(-)

Kt qu th nghim cho thy, hp cht HM1 khng 03 chng vi sinh vt

kim nh l P. aeruginosa v S. aureus, hp cht HM2 khng 04 chng l P.
aeruginosa, B. subtilis, F. oxysporum v S. cerevisiae, HM3 khng 03 chng l

452

E. coli, B. subtilis, v S. cerevisiae. Trong , trc khun m xanh Pseudomonas

aeruginosa (thuc nhm vi khun Gram (-) l vi khun gy khng thuc khng
sinh mnh v khng hu ht cc thuc khng sinh th h c) v t cu vng
Staphylococcus aureus (thng gy nhim trng v bi nhim nng ngoi da
cng nh mt s c quan v thng c mt trong cc trng hp nhim trng
mu v cng l chng khng thuc vi nhiu thuc khng sinh). Hp cht HM2
v HM3, thuc nhm cht sulfuno glycolipit, c cu trc c th vi s c mt
ca d t lu hunh (S) trong phn t khng nhng gi nhiu chc nng quan
trng trong c th sinh vt m cn th hin nhiu hot tnh qu bu. Chng c
phn lp vi hm lng cao trong loi hi min G. varius sinh sng ph bin ti
cc vng bin Vit Nam. iu ny t c s khoa hc cho cc nghin cu tip
theo nhm to ra cc dc phm mi trong iu tr cc bnh him ngho ng
thi s dng mt cch hp l v c hiu qu ngun dc liu bin v cng qu
gi ca t nc.
V.2.6. Cc cng trnh cng b
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc
loi hi min Gellius varius, chng ti bin son v ng 02 cng trnh trn
tp ch chuyn ngnh quc gia l:
(1) Nguyn Xun Cng, Phan Vn Kim v Chu Vn Minh, Cc hp cht glycolipit t
loi hi min cnh xanh Gellius varius sinh sng ti vng bin Vit Nam. Tp ch Khoa
hc v Cng ngh, 2007, 45(2), 41-47.
(2) Nguyn Xun Cng, Nguyn Hi ng, Cng Thung, Trn Thu Hng, Nguyn
Th Kim Thy, Phan Vn Kim v Chu Vn Minh, Isoprenoids from the sponge
Gellius varius living in Vietnamese sea. Tp ch Ha hc, 2007, 45(6), 776-780.

Cc kt qu trn c s dng trong mt lun n Thc s Ha hc v bo

v thnh cng vi ti Nghin cu thnh phn ha hc v hot tnh gy c t
bo ca loi hi min Gellius varius, ThS. Nguyn Th Kim Thy.
Bng tng hp cc cht phn lp c t loi hi min
cnh xanh Gellius varius
HO
OH
4"

6"
5"

6'

OH
3"

CH2SO3H

1"

2"

OH OH
4'

5'

6'
5'

CH2

2
1' HC

OH
3'

2'

OH

4'

OH

OH

CH2 O

1'"

O
1'

OH
2'
3'

OH

16'"

CH2
2

HC

1'"

(CH2)14CH3
1

HM1: 1-O-Palmitoyl-3-O-[-Dgalactopyranosyl-(16)--Dgalactopyranosyl]-sn-glycerol

CH2 O

16'"

(CH2)14CH3

O 1"
16"
C (CH2)14CH3
O

HM2: (2S)1,2-Di-O-Palmitoyl-3-O-(6sulfo--D-quinovopyranosyl) glycerol

453

6'

CH2SO3Na
5'
4'

21

OH
2'

OH

3'

HC

OH

1
1"

CH2 O

16"

(CH2)14CH3

20

18

HM4: Cholesterol

22

21

21

26

25
27

17

15

7 O

10

HO

27

13

26
25

17

11
19

10

22
20

18

13
8

HO

HO

HM3: Mui natri ca (2S)1-OHexadecanoyl-3-O-(6-sulfo--Dquinovopyranosyl) Glycerol

19

25

26

19

11

24

17

CH2
2

OH

21

20
18

1'

15

OH

OH

HM5: 5,8-Epidioxycholest-6-en-3-ol
21
18

HM6: Cholest-7-ene-3,5,6-triol

22

26

20
25

O
8

HO

11

13

19
8

10

27

17

11

15

20

17

14

OH
OCH3

HM8: Metyl este ca axit

(5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic

HM7: 6-Methoxycholest-7-ene-3,5diol

HO
17

18

19

O
5'

8'

15

11

OH
2

20
1

1'

OH

16

20'
11'

14'

HM10: Glycerol 1-(5Z,8Z,11Z,14Zeicosatetraenoate)

HM9: Phytol

454

V.3. Kt qu nghin cu ho hc v hot tnh sinh hc ca

loi hi min Ianthella sp. (ianthellidae)
V.3.1. M u
Mu hi min
Ianthella sp. c thu
thp ti Nam Yt,
Qun o Trng Sa,
tnh Khnh Ha vo
thng 03 nm 2007 v
c TS. Cng
Thung v cc cng
s, Vin Ti nguyn
v Mi trng Bin,
Vin KH&CN Vit
Nam gim nh tn
khoa hc.

Hi min Ianthella sp.

- M t: C th c dng qut, mu sc thay i t mu vng nht, mu vng cam
m cho ti mu tm sm, khi tip xc vi khng kh thng chuyn mu tm sm

Ianthella l mt chi hi min thuc h Ianthellidae, sinh sng kh ph

bin ti cc vng bin Vit Nam. Hin nay nc ta hon ton cha c cng
trnh khoa hc no c cng b v thnh phn ha hc v hot tnh sinh hc ca
cc loi thuc chi ny. Cc nghin cu trn th gii cho thy, cc dng dn xut
bastadin l thnh phn ha hc chnh ca cc loi thuc chi Ianthella. Hp cht
bastadin u tin c phn lp vo nm 1981, bastadin 1 (1) [269]. Nm 1990,
nhm nghin cu ca tc gi Miao SC cng b s phn lp, xc nh cu trc v
hott tnh gy c t bo ca bastadin 8 (2) v bastadin 9 t loi I. basta [270].
n nm 1993, nhm nghin cu ca tc gi Gulavita NK cng b hp cht 34sulfatobastadin 13 (3) th hin hot tnh c ch th quan endothelin A t loi hi
min Ianthella sp. [271]. Chn hp cht mi, hemibastadins 1-3 (4),
hemibastadinols 1-3 (5) [272], bastadin 20 (6), 15,34-O-disulfatobastadin 7 v
10-O-sulfatobastadin 3 (7) [273] tip tc c cng b t loi I. basta vo nm
1996. Nm 2001, hai hp cht mi v rt him, iantherans A (8) v B, c phn
lp t loi hi min Australia Ianthella sp.. Chng th hin hot tnh c ch enzim
Na,K-ATPase v nhiu enzym khc [274]. Nm 2007, nhm nghin cu ca tc
gi Greve H cng b 3 hp cht iantheran mi, iso-iantheran A, 8-carboxy-isoiantheran A v iso-iantheran B t loi I. quadrangulata [275]. Gn y nht, mt
hp cht mi bastatin 24 (9) tip tc c phn lp t loi I. quadrangulata [276].
Hp cht ny c ch chn lc 5 trong s 36 dng t bo ung th c nghin cu.
Di y, chng ti gii thiu mt s hp cht tiu biu c phn lp
t loi Ianthella sp.

455

OH

Br

Br

NOH

HO

CH 2 CH 2 NHCOCCH 2

C
O

OH
O

Br

HO

Br

Br

HO

OH

H
N

Br

Br OH

Br

CH2 CH2 NHCOCCH2

Br

O
C

H
N

NOH

1 [269]

HO

2 [270]
N

H
N

OH

OH

Br

C
O

HO

O
Br
H
N

N
H

Br

Br
OSO3Na
Br

Br

HO

O
C

HO

N
H

Br

OH

OH

3 [271]
H
N

4 [272]

Br

5'

Br
OH

5'

OH

5 [272]
OH

Br

HO

34

Br

Br

OH

NOH
CH2CCONHCH2CH2

HO
HO

Br
28

Br
N
H

Br
N

Br

OH

7 [273]
N

H
N

Br

Br

OH

C
O

OH

Br

O
Br

OSO3H

HO

1'

Br

HO3SO

Br

Br
HO

OH

CH2CCONHCH2CH2
NOH

6 [273]

OSO3H

10

O
C
N
H

Br

OH
HO N

9 [276]

8 [274]

456

Br
OH
Br

V.3.2. Phn lp cc hp cht

Hi min Ianthella sp. (10 kg ti) c ra sch, xay nh v chit bng
metanol (3 ln) bng thit b chit siu m. Dch metanol ca cc ln chit c
lc, gom li v ct loi dung mi nhit khng qu 60oC di p sut gim
thu c 70 g dch cn dch metanol. Cn dch ny c ha vo nc v chit
phn b ln lt vi cc dung mi hexan v etyl axetat theo t l 1/1 (3 ln, mi
ln 6 lit) thu c cc cn dch tng ng l hexan 10 g (k hiu: IH), EtOAc 9.3
g (k hiu IE) v nc 50 g (k hiu IN) (S V.3.2.a).
Cn chit EtOAc: IE (9.3 g) c tin hnh chy ct th trn sc k ct
pha thng gradient nng metanol trong cloroform thu c 4 phn on nh
hn l IE1-IE4. Cc phn on IE1, IE2 v IE3 c tip tc phn tch trn sc
k ct pha thng vi h dung mi ra gii thch hp thu c cc hp cht IS1
n IS5 nh c ch ra trn S V.3.2.b.

Hi min Ianthella sp. ti

(10 kg)
- Chit MeOH, ct loi dung mi di p sut
gim
B sung nc

IM - Cn chit MeOH (70 g)

B sung hexan
hexan: nc 1/1

Lp nc
B sung EtOAc

Cn hexan
IH (10 g)

EtOAc: nc 1/1

Lp nc

Cn nc
IN (50 g)

Cn EtOAc
IE (9.3 g)

S V.3.2.a: S chit phn on mu hi min

457

IE (9.3 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

IE1 (0.5 g)

Silica gel CC
CM 30/1

IE2 (1 g)

IE3 (1.8 g)

IE4 (6 g)

YMC RP-18 CC
MWA 5/1/3

IS1

IS3
7.5 mg

90 mg

S V.3.2.b: S phn lp cc hp cht t cn chit EtOAc

IE3 (1.8 g)
Silica gel CC
HE 5/1

IE3A (0.2 g)

IE3B (0.6 g)

IE3C (1.0 g)

RP CC (YMC ODS-A)
MeOH-H2O-Acetone (5:1:3)

IS4
10 mg

IE3C1 (100 mg)

Kt tinh li
trong axeton

IS2
12 mg

IE3C2 (0.5 g)

RP CC (YMC ODS-A))
MeOH-H2O-Acetone (5:1:3)

IE3C3 (0.4 g)

CC (silica gel,)
n-hexane:EtOAc=3/2

IS5
(10mg)

S V.3.2.c: S phn lp cc hp cht t cn chit cloroform

458

V.3.3. Hng s vt l v d kin ph ca cc hp cht

V.3.3.1. Hp cht IS1: petrosterol
Cht bt mu trng, C29H48O (M = 412).
1
H-NMR (500 MHz, CDCl3) (ppm): 1,85 (1H, m, Ha-1), 1,05 (1H,m, Hb-1),
1,78 (2H, m, H-2), 3,42 (1H, m, H-3), 1,95-2,20 (2H, m, H-4), 5,30 (1H, br d, J = 5,5
Hz, H-6), 1,95 (1H, m, Ha-7), 1,54 (1H, m, Hb-7), 1,42-1,50 (2H, m, H-8), 0,92 (1H, m,
H-9), 1,32-1,40 (2H, m, H-11), 2,05 (1H, ddd, J = 12,0, 5,0, 2,0 Hz, H-12), 0,95-1,0
(1H, m, H-14), 1,82 (1H, m, Ha-15), 1,28 (1H, m, Hb-15), 1,58 (1H, m, Ha-16), 1,09
(1H, m, Hb-16), 1,10 (1H, m, H-17), 0,68 (3H, s, H-18), 1,0 (3H, s, H-19), 1,36 (1H, m,
H-20), 0,92 (3H, d, J = 7,0 Hz, H-21), 1,50 (1H, m, Ha-22), 1,00 (1H, m, Hb-22), 1,35
(1H, m, Ha-23), 1,25 (1H, m, Hb-23), 0,58 (1H, m, H-24), 0,12 (1H, m, H-25), 0,45 (1H,
m, H-26), 0,12 (1H, m, Ha-27), 0,05 (1H, m, Hb-27), 1,03 (3H, d, J = 6,0 Hz, H-28) v
0,88 (3H, d, J = 6,0 Hz, H-29).
13
C-NMR (125 MHz, CDCl3) (ppm): 38,18(C-1), 31,94(C-2), 72,02(C-3),
42,65(C-4), 141,78(C-5), 122,29(C-6), 32,84(C-7), 32,73(C-8), 51,19(C-9), 37,36(C-10),
21,91(C-11), 40,73(C-12), 43,16(C-13), 57,73(C-14), 29,07(C-15), 25,06(C-16),
57,10(C-17), 12,42(C-18), 19,56(C-19), 36,85(C-20), 19,26(C-21), 34,37(C-22), 34,8(C23), 39,68(C-24), 28,19(C-25), 13,59(C-26), 12,18(C-27), 19,90(C-28) v 20,35(C-29).
V.3.3.2. Hp cht IS2: iansterol A (cht mi)
Cht bt mu trng, C29H44O2 (M = 424), ph khi lng FAB-MS: m/z 425
[M+H]+, ph khi lng phn gii cao HR-FAB-MS: m/z 425,3397 [M+H]+ (tnh ton l
thuyt cho C29H45O2 l 425,3375).
1
H-NMR (500 MHz, CDCl3) (ppm): 2,18 (1H, m, Ha-1), 1,90 (1H, m, Hb-1),
2,54 (1H, m, Ha-2), 2,50 (1H, m, Hb-2), 6,18 (1H, s, H-4), 2,68 (1H, dd, J = 16,0, 4,0
Hz, Ha-7), 2,06 (1H, m, Hb-7), 1,90 (1H, m, H-8), 1,40 (1H, m, H-9), 1,65 (1H, m, Ha11), 1,51 (1H, m, Hb-11), 2,11 (1H, m, Ha-12), 1,28 (1H, m, Hb-12), 1,20 (1H, m, H-14),
1,63 (1H, m, Ha-15), 1,18 (1H, m, Hb-15), 1,90 (1H, m, Ha-16), 1,33 (1H, m, Hb-16),
1,20 (1H, m, H-17), 0,73 (3H, s, H-18), 1,17 (3H, s, H-19), 1,38 (1H, m, H-20), 0,94
(3H, d, J = 6,8 Hz, H-21), 1,50 (1H, m, Ha-22), 1,22 (1H, m, Hb-22), 1,25-1,75 (2H, m,
H-23), 0,61 (1H, m, H-24), 0,15 (1H, m, H-25), 0,46 (1H, m, H-26), 0,14 (1H, m, Ha27), 0,07 (1H, m, Hb-27), 1,02 (3H, d, J = 6,4 Hz, H-28) v 0,90 (3H, d, J = 6,4 Hz, H29).
13
C-NMR (125 MHz, CDCl3) (ppm): 35,76 (C-1), 34,19 (C-2), 199,70 (C-3),
125,67 (C-4), 161,28 (C-5), 202,55 (C-6), 47,04 (C-7), 34,43 (C-8), 51,20 (C-9), 40,03
(C-10), 21,11 (C-11), 39,36 (C-12), 42,75 (C-13), 56,17 (C-14), 24,19 (C-15), 28,22 (C16), 56,79 (C-17), 12,12 (C-18), 17,73 (C-19), 36,05 (C-20), 18,86 (C-21), 33,61 (C-22),
34,13 (C-23), 38,92 (C-24), 27,57 (C-25), 13,02 (C-26), 11,81 (C-27), 19,36 (C-28) v
20,08 (C-29).
V.3.3.3. Hp cht IS3: iansterol B (cht mi)
Cht bt mu trng, C29H48O2 (M = 428), ph khi lng FAB-MS: m/z 429
+
[M+H] , ph khi lng phn gii cao HR-EI-MS: m/z 428,3656 [M]+ (tnh ton l
thuyt cho C29H48O2 l 428,3654).

459

H-NMR (500 MHz, CDCl3) (ppm): 1,70 (1H, m, Ha-1), 1,36 (1H, m, Hb-1),
1,92 (1H, m, Ha-2), 1,62 (1H, m, Hb-2), 3,90 (1H, m, H-3), 2,08 (1H, m, Ha-4), 1,31
(1H, m, Hb-4), 2,92 (1H, d, J = 4,4 Hz, H-6), 1,93 (1H, m, Ha-7), 1,50 (1H, m, H-b7),
1,38 (1H, m, H-8), 1,26 (1H, m, H-9), 1,37 (1H, m, Ha-11), 1,25 (1H, m, Hb-11), 1,95
(1H, m, Ha-12), 1,10 (1H, m, Hb-12), 1,07 (1H, m, H-14), 1,55 (1H, m, Ha-15), 1,03
(1H, m, Hb-15), 1,82 (1H, m, Ha-16), 1,26 (1H, m, Hb-16), 0,95 (1H, m, H-17), 0,61
(3H, s, H-18), 1,07 (3H, s, H-19), 1,33 (1H, m, H-20), 0,90 (3H, d, J = 6,5 Hz, H-21),
1,46 (1H, m, Ha-22), 0,98 (1H, m, Hb-22), 1,34 (1H, m, Ha-23), 1,23 (1H, m, Hb-23),
0,59 (1H, m, H-24), 0,14 (1H, m, H-25), 0,45 (1H, m, H-26), 0,14 (1H, m, Ha-27), 0,08
(1H, m, Hb-27), 1,00 (3H, d, J = 6,0 Hz, H-28) v 0,88 (3H, d, J = 6,5 Hz, H-29).
1

C-NMR (125 MHz, CDCl3) (ppm): 32,38 (C-1), 31,07 (C-2), 68,78 (C-3),
39,84 (C-4), 65,71 (C-5), 59,31 (C-6), 28,81 (C-7), 29,88 (C-8), 42,55 (C-9), 34,85 (C10), 20,64 (C-11), 39,84 (C-12), 42,32 (C-13), 56,04 (C-14), 24,05 (C-15), 28,07 (C-16),
56,85 (C-17), 11,87 (C-18), 15,92 (C-19), 35,90 (C-20), 18,62 (C-21), 33,44 (C-22),
33,93 (C-23), 39,40 (C-24), 27,36 (C-25), 12,77 (C-26), 11,56 (C-27), 19,13 (C-28) v
19,85 (C-29).
13

V.3.3.4. Hp cht IS4: aragusterol B

Cht bt mu trng, C27H48O3 (M = 444).
1
H-NMR (CDCl3, 500 MHz) (ppm): 2,04 (1H, ddd, J = 13,0, 6,5, 4,5 Hz, H-1),
1,36 (1H, m, H-1), 2,42 (1H, dt, J = 3,5, 6,5 Hz, H-2), 2,33 (1H, m, H-2), 2,20 (1H, t, J
= 14,5 Hz, H-4), 2,10 (1H, dq, J = 14,5, 1 Hz, H-4), 1,53 (1H, m, H-5), 1,3-1,4 (2H, m,
H-6), 1,72 (1H, m, H-7), 0,88 (1H, m, H-7), 1,38 (1H, m, H-8), 0,83 (1H, m, H-9), 1,76
(1H, m, H-11), 1,36 (1H, m, H-11), 3,37 (1H, dd, J = 11,0, 4,5 Hz, H-12), 1,00 (1H, m,
H-14), 1,70 (1H, m, H-15), 1,22 (1H, m, H-15), 1,72 (1H, m, H-16), 1,50 (1H, m, H16), 1,65 (1H, t, J = 6,5 Hz, H-17), 0,86 (3H, s, H-18), 1,02 (3H, s, H-19), 1,16 (3H, s,
H-21), 1,55 (1H, m, H-22), 1,32 (1H, m, H-22), 1,82 (1H, brt, J = 10,5 Hz, H-23), 1,58
(1H, m, H-23), 0,68 (1H, q, J = 6,5 Hz, H-24), 0,18 (1H, m, H-25), 0,50 (1H, m, H-26),
0,18 (1H, m, H-27), 0,12 (1H, m, H-27), 1,03 (3H, d, J = 6,0 Hz, H-28) v 0,94 (3H, d,
J = 6,5 Hz, H-29).
13
C-NMR (CDCl3, 125 MHz) (ppm): 38,6 (C-1), 38,4 (C-2), 211,2 (C-3), 44,8
(C-4), 46,8 (C-5), 29,1 (C-6), 31,3 (C-7), 33,9 (C-8), 52,7 (C-9), 35,8 (C-10), 29,4 (C11), 77,9 (C-12), 48,9 (C-13), 54,8 (C-14), 23,6 (C-15), 25,2 (C-16), 64,4 (C-17), 9,8 (C18), 11,6 (C-19), 75,9 (C-20), 28,9 (C-21), 34,1 (C-22), 31,9 (C-23), 39,2 (C-24), 27,3
(C-25), 13,1 (C-26), 11,8 (C-27), 19,4 (C-28) v 20,5 (C-29).
V.3.3.5. Hp cht IS5: iansterol C (cht mi)
Cht bt mu trng, ph khi lng FAB-MS: m/z 461 [M-CH2O+H]+ (C31H54O4,
M = 490).
H-NMR (500 MHz, CDCl3) (ppm): 1,58 (1H, m, Ha-1), 1,32 (1H, m, Hb-1),
1,90 (1H, m, Ha-2), 1,45 (1H, m, Hb-2), 1,63 (1H, m, H-4), 1,30 (1H, m, H-4), 1,31 (1H,
m, H-5), 1,29 (1H, m, Ha-6), 1,18 (1H, m, Hb-6), 1,68 (1H, m, Ha-7), 0,88 (1H, m, Hb-7),
1,32 (1H, m, H-8), 0,83 (1H, m, H-9), 1,73 (1H, m, Ha-11), 1,27 (1H, m, Hb-11), 3,36
1

460

(1H, dd, J = 11,0, 4,5 Hz, H-12), 0,96 (1H, m, H-14), 1,68 (1H, m, H-15), 1,20 (1H, m,
H-15), 1,71 (1H, m, Ha-16), 1,49 (1H, m, Hb-16), 1,65 (1H, m, H-17), 0,84 (3H, s, H18), 0,81 (3H, s, H-19), 1,16 (3H, s, H-21), 1,55 (1H, m, Ha-22), 1,32 (1H, m, Hb-22),
1,58 (1H, m, Ha-23), 1,30 (1H, m, Hb-23), 0,66 (1H, q, J = 6,5 Hz, H-24), 0,16 (1H, m,
H-25), 0,50 (1H, m, H-26), 0,16 (1H, m, Ha-27), 0,11 (1H, m, Hb-27), 1,03 (3H, d, J =
6,0 Hz, H-28), 0,93 (3H, d, J = 6,5 Hz, H-29), 3,14 (3H, s, OMe) v 3,19 (3H, s, OMe).
C-NMR (125 MHz, CDCl3) (ppm): 35,18 (C-1), 28,60 (C-2), 100,60 (C-3),
35,61 (C-4), 42,60 (C-5), 28,62 (C-6), 31,59 (C-7), 34,07 (C-8), 52,95 (C-9), 35,94 (C10), 29,31 (C-11), 78,21 (C-12), 48,87 (C-13), 55,07 (C-14), 23,68 (C-15), 25,29 (C-16),
64,40 (C-17), 9,81 (C-18), 11,76 (C-19), 75,89 (C-20), 28,93 (C-21), 34,15 (C-22),
31,94 (C-23), 39,26 (C-24), 27,31 (C-25), 13,12 (C-26), 11,78 (C-27), 19,40 (C-28),
20,45 (C-29), 47,69 v 47,73 (OMe).
13

461

V.3.4. Xc nh cu trc ha hc ca cc hp cht

V.3.4.1. Hp cht IS1: petrosterol
Hp cht IS1 phn lp c di dng tinh th hnh kim mu trng. Ph
1
H-NMR ca IS1 c dng c trng ca mt hp cht steroit. Trong , 01 proton
olefin v 01 proton oximetin c xc nh ln lt bi cc tn hiu cng hng
ti 5,30 (1H, br d, J = 5,5 Hz, H-6) v 3,42 (1H, m, H-3). Cc tn hiu cng
hng ti 0,68 (3H, s, H-18), 1.00 (3H, s, H-19), 0,92 (3H, d, J = 7,0 Hz, H21), 1,03 (3H, d, J = 6,0 Hz, H-28) v 0,88 (3H, d, J = 6,0 Hz, H-29) khng nh
s tn ti ca 2 nhm metyl bc ba v 3 nhm metyl bc hai. S xut hin ca
cu trc vng cyclopropan c chng minh bi cc tn hiu proton cng hng
ti 0,12 (1H, m, H-25), 0,45 (1H, m, H-26), 0,12 (1H, m, Ha-27) v 0,05 (1H,
m, Hb-27).

Hnh V.3.4.1.a. Ph 1H-NMR ca IS1

29
21

22

18

19

12
11

13
14

2
3

HO

10
5

20

26

24

28
23

25
27

17
16
15

8
7

Hnh V.3.4.1.b. Cu trc ha hc ca IS1

Ph 13C-NMR ca IS1 xut hin cc tn hiu ca 29 cacbon. Trong ,
mt nhm oximetin v mt lin kt i b th 3 v tr c khng nh bi cc tn
hiu cng hng ti 72,02 (CH, C-3) v 141,78 (C, C-5)/122,29 (CH, C-6).

462

Nm nhm metyl c c trng bi cc tn hiu cng hng ti 12,42 (C-18),

19,56 (C-19), 19,26 (C-21), 19,90 (C-28) v 20,35 (C-29).

Hnh V.3.4.1.c. Ph 13C-NMR ca IS1

Hnh V.3.4.1.d. Ph 13C-NMR dn rng ca IS1

463

Tt c cc s liu chuyn dch ha hc ca cacbon c gn vi cc s

liu proton tng ng thng qua ph HMQC. Kt qu chi tit c tng hp
trong bng V.3.4.1.

Hnh V.3.4.1.e. Ph HMQC ca IS1

Bng V.3.4.1. Kt qu ph NMR ca IS1
C

Ca, b

37,5

38,18

2
3
4
5
6

31,9
72,0
42,5
141,0
122,0

31,94
72,02
42,65
141,78
122,29

32,1

32,84

8
9
10
11
12
13
14
15

32,1
50,3
36,7
21,3
40,0
42,4
57,0
28,5

32,73
51,19
37,36
21,91
40,73
43,16
57,73
29,07

Ha, c
dng pic (J = Hz)
1,85 (1H, m)
1,05 (1H, m)
1,78 (2H, m)
3,42 (1H, m)
1,95-2,20 (2H, m)
5,30 (1H, br d, J = 5,5 Hz)
1,95 (1H, m)
1,54 (1H, m)
1,42-1,50 (2H, m)
0,92 (1H, m)
1,32-1,40 (2H, m)
2,05 (1H, ddd, J = 12,0, 5,0, 2,0 Hz)
0,95-1,0 (1H, m)
1,82 (1H, m)
464

16

24,5

25,06

17
18
19
20
21

56,3
12,1
19,4
36,1
18,9

57,10
12,42
19,56
36,85
19,26

22

33,7

34,37

23

34,1

34,8

24
25
26

38,9
27,6
13,0

39,68
28,19
13,59

27

19,4

12,18

28
29

19,4
20,1

19,90
20,35

1,28 (1H, m)
1,58 (1H, m)
1,09 (1H, m)
1,10 (1H, m)
0,68 (3H, s)
1,00 (3H, s)
1,36 (1H, m)
0,92 (3H, d, J = 7,0 Hz)
1,50 (1H, m)
1,0 (1H, m)
1,35 (1H, m)
1,25 (1H, m)
0,58 (1H, m)
0,12 (1H, m)
0,45 (1H, m)
0,12 (1H, m)
0,05 (1H, m)
1,03 (3H, d, J = 6,0 Hz)
0,88 (3H, d, J = 6,0 Hz)

o trong CDCl3+MeOD, b125 MHz, c500 MHz, #s liu ca petrosterol [277].

Hnh V.3.4.1.f. Ph 1H-1H COSY ca IS1

Phn tch chi tit cc tng tc trn ph 1H-1H COSY kt hp vi cc
tng tc xa trn ph HMBC cho php xc nh v tr ca nhm oximetin ti C-3,
lin kt i ti C-5/C-6, vng propan ti C-25/C-26 cng nh cu trc ca ton b
phn t nh c ch ra trn hnh V.3.4.h.

465

Hnh V.3.4.1.g. Ph HMBC ca IS1

HMBC
COSY

HO

Hnh V.3.4.1.h. Cc tng tc HMBC (HC) v COSY chnh ca IS1

T tt c cc phn tch nu trn, cng vi s ph hp hon ton v s liu
ph NMR ca IS1 so vi cc s liu tng ng c cng b [277] cho php
xc nh cu trc ha hc ca IS1 l 26,27-cyclo-26-methylergost-5-en-3-ol
(petrosterol).
V.3.4.2. Hp cht IS2: iansterol A (cht mi)
Hp cht IS2 c phn lp di dng cht bt mu trng. Cng thc phn
t ca n c xc nh l C29H44O2 (M = 424) bng cc d kin ph khi lng
bn ph nguyn t nhanh (FAB-MS ti m/z 425 [M+H]+) v ph khi lng phn
gii cao (HR-FAB-MS ti m/z 425,3397 [M+H]+, tnh ton l thuyt cho
C29H45O2 425,3375).

466

Hnh V.3.4.2.a. Ph 1H-NMR ca IS2

Ph 1H-NMR ca IS2 c dng tng t nh ca IS1. iu ny cho thy
hai hp cht c cu trc tng t nhau. S khc bit d nhn thy nht trn ph
1
H-NMR ca IS2 so vi IS1 l s mt i tn hiu ca nhm oximetin ti C-3.

Hnh V.3.4.2.b. Ph 13C-NMR ca IS2

467

Hnh V.3.4.2.c. Ph 13C-NMR dn rng ca IS2

Hnh V.3.4.2.e. Ph HSQC ca IS2

468

29
21

22

18

20

12
19

11

27

15

9
8

10
5

25

16

14
2

28
23

17

13

26

24

7
6

Hnh V.3.4.2.d. Cu trc ha hc ca IS2

Ph 13C-NMR ca IS2 c dng tng t nh ca IS1 vi s xut hin cc
tn hiu ca 29 cacbon. S khc bit d nhn thy nht trn ph 13C-NMR ca IS2
so vi IS1 l s mt i tn hiu ca nhm oximetin v s xut hin thm cc tn
hiu ca 2 nhm ketn ti 199,70 (C-3) v 202,55 (C-6). Tn hiu ca lin kt
i b th 3 v tr ti 125,67/161,28 gi rng v tr ca lin kt i trong phn
t ca IS2 b thay i so vi IS1.
Bng V.3.4.2. Kt qu ph NMR ca IS2
C
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

C#
35,5

Ha, c
dng pic (J = Hz)

Ca, b
35,76

34,0
199,5
125,5
162,8
202,3
46,8

34,19
199,70
125,67
161,28
202,55

34,2
50,9
39,0
20,8
39,8
41,9
56,6
23,9
29,7
55,8
12,0
17,5
39,0
20,8

34,43
51,20
40,03
21,11
39,36
42,75
56,17
24,19
28,22
56,79
12,12
17,73
36,05
18,86
33,61

47,04

2,18 (1H, m)
1,90 (1H, m)
2,54 (1H, m); 2,50 (1H, m)
6,18 (1H, s)

2,68 (1H, dd, J = 16,0, 4,0 Hz)

2,06 (1H, m)
1,90 (1H, m)
1,40 (1H, m)
1,65 (1H, m); 1,51 (1H, m)
2,11 (1H, m); 1,28 (1H, m)
1,20 (1H, m)
1,63 (1H, m); 1,18 (1H, m)
1,90 (1H, m); 1,33 (1H, m)
1,20 (1H, m)
0,73 (3H, s)
1,17 (3H, s)
1,38 (1H, m)
0,94 (3H, d, J = 6,8 Hz)
1,50 (1H, m); 1,22 (1H, m)

469

23
24
25
26
27
28
29

34,13
38,92
27,57
13,02
11,81
19,36
20,08

1,25-1,75 (2H, m)
0,61 (1H, m)
0,15 (1H, m)
0,46 (1H, m)
0,14 (1H, m); 0,07 (1H, m)
1,02 (3H, d, J = 6,4 Hz)
0,90 (3H, d, J = 6,4 Hz)

o trong CDCl3, b125 MHz, c500 MHz, #s liu ca (20R)-22E-cholesta-4,22-diene-3,6-dione [278].

Hnh V.3.4.2.f. Ph HMBC ca IS2

HMBC
COSY

O
O

Hnh V.3.4.2.g. Cc tng tc HMBC (HC) chnh ca IS2

V tr ca 2 nhm ktn (ti C-3 v C-6) v lin kt i (ti C-4/C-5) c
xc nh bi s ph hp hon ton v s liu phn khung steroit ca IS2 so vi
cc s liu c cng b ca hp cht (20R)-22E-cholesta-4,22-diene-3,6dione [278].
470

Hnh V.3.4.2.f. Ph khi lng FAB-MS ca IS2

Hnh V.3.4.2.f. Ph khi lng phn gii cao HR-FAB-MS ca IS2

T tt c cc phn tch nu trn, cu trc ha hc ca IS2 c xc nh l
26,27-cyclo-26-methylergost-4-en-3,6-dione. y l mt hp cht c cu trc
mi, ln u tin c phn lp t thin nhin v c chng ti t tn l
iansterol A.
V.3.4.3. Hp cht IS3: iansterol B (cht mi)
Hp cht IS3 cng c phn lp di dng cht bt mu trng. T cc kt
qu ph NMR, ph khi lng (FAB-MS ti m/z 429 [M+H]+) v ph khi lng
phn gii cao (HR-EI-MS ti m/z 428.3656, tnh ton l thuyt cho C29H48O2 l
428.3654) cho php xc nh cng thc phn t ca IS3 l C29H48O2.

471

Hnh V.3.4.3.a. Ph 1H-NMR ca IS3

Hnh V.3.4.3.b. Ph 13C-NMR ca IS3

Bng V.3.4.3. Kt qu ph NMR ca IS3

C

C#

C$

Ca, b

1
2
3
4
5
6
7
8
9

37,21
31,03
69,44
42,26
62,91
63,72
32,59
29,74
51,31

32,36
31,01
68,66
39,81
65,73
59,29
28,73
29,84
42,51

32,38
31,07
68,78
39,84
65,71
59,31
28,81
29,88
42,55
472

Ha, c
dng pic (J = Hz)
1,70 (1H, m), 1,36 (1H, m)
1,92 (1H, m), 1,62 (1H, m)
3,90(1H, m)
2,08 (1H, m), 1,31 (1H, m)
2,92 (1H, d, J = 4,4)
1,93 (1H, m), 1,50 (1H, m)
1,38 (1H, m)
1,26 (1H, m)

10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29

34,82
21,97
39,79
42,21
56,20
24,17
28,15
56,07
11,74
17,03
36,05
18,71
33,85
25,98

35,09
20,59
39,62
42,29
56,80
24,02
28,07
55,72
11,82
15,89
36,09
18,66
34,10
26,01

34,85
20,64
39,84
42,32
56,04
24,05
28,07
56,85
11,87
15,92
35,90
18,62
33,44
33,93
39,40
27,36
12,77
11,56
19,13
19,85

1,37 (1H, m), 1,25 (1H, m)

1,95 (1H, m), 1,10 (1H, m)
1,07 (1H, m)
1,55 (1H, m), 1,03 (1H, m)
1,82 (1H, m), 1,26 (1H, m)
0,95 (1H, m)
0,61 (3H, s)
1,07 (3H, s)
1,33 (1H, m)
0,90 (3H, d, J = 6,5)
1,46 (1H, m); 0,98 (1H, m)
1,34 (1H, m); 1,23 (1H, m)
0,59 (1H, m)
0,14 (1H, m)
0,45 (1H, m)
0,14 (1H, m); 0,08 (1H, m)
1,00 (3H, d, J = 6,0)
0,88 (3H, d, J = 6,5)

o trong CDCl3, b125 MHz, c500 MHz, #s liu ca 5,6-epoxy-5-stigmastan-3-ol [279], $s liu ca
5,6-epoxy-5-stigmastan-3-ol [279],

29
21

22

18
12
19

11

13
14

2
3

HO

20
17
16

26

24

28
23

25
27

15

10

Hnh V.3.4.3c. Cu trc ha hc ca IS3

Ph 1H-NMR ca IS3 c dng tng t nh ca IS1 v IS2. im khc
bit ln nht trn ph 1H-NMR ca IS3 so vi IS1 l s mt i tn hiu ca
proton olefin v thay vo l tn hiu ca mt nhm oximetin. Trn ph 13CNMR ca IS3 xut hin cc tn hiu cng hng ca 29 cacbon. So vi IS1, tn
hiu ca lin kt i b mt i v thay vo l tn hiu ca mt nhm
oximetin ( 59,31) v mt cacbon bc bn ni vi nguyn t xy ( 65,71). iu
ny gi cho s c mt ca mt cu epoxy ti v tr C-5/C-6.

473

Hnh V.3.4.3.d. Ph khi lng phn gii cao HR-EI-MS ca IS3

Cu hnh ca cu epoxy c xc nh l 5,6 bng cch so snh s liu
ph C-NMR ca phn khung steroit c bit l ti hai v tr C-5 v C-6 ca IS3
(ti 65,71/59,31) vi cc s liu tng ng ca 5,6-epoxy-5-stigmastan-3-ol
(ti 65,73/59,29) [279] v 5,6-epoxy-5-stigmastan-3-ol (ti 62,91/63,72)
[279]. T tt c cc phn tch nu trn, cu trc ha hc ca IS3 c xc nh l
5,6-epoxy-26,27-cyclo-26-methylergost-3-ol. y l mt hp cht mi v
c chng ti t tn l iansterol B.
13

V.3.4.4. Hp cht IS4: aragusterol B

Hp cht IS4 nhn c di dng cht bt mu trng. Ph 1H-NMR ca
IS4 c trng cho mt hp cht steroit, trong xut hin cc tn hiu ca mt
nhm oximetin ti 3,37 (1H, dd, J = 11,0 v 4,5 Hz, H-12). Ba nhm metyl bc
ba v hai nhm metyl bc hai c xc nh ln lt bi cc tn hiu cng hng
ti 0,86 (3H, s, H-18), 1,02 (3H, s, H-19), 1,16 (3H, s, H-21), 1,03 (3H, d, J =
6,0 Hz, H-28) v 0,94 (3H, d, J = 6,5 Hz, H-29).

Hnh V.3.4.4.a. Ph 1H-NMR ca IS4

474

Ph 13C-NMR xut hin cc tn hiu ca mt steroit 29 cacbon. Trong ,

mt nhm ktn, mt nhm oximetin v mt cacbon bc bn ni vi nguyn t
xy c xc nh ln lt bi cc tn hiu cng hng ti 211,2 (C-3), 77,9 (C12) v 75,9 (C-20). Cc tn hiu ti 9,8 (CH3, C-18), 11,6 (CH3, C-19), 28,9
(CH3, C-21), 19,4 (CH3, C-28) v 20,5 (CH3, C-29) cho php xc nh s c mt
ca 5 nhm metyl trong phn t ca IS4.
29

21

HO

18

12
19

11

13
14

2
3

22

HO
20

26

24

28
23

25
27

17
16
15

10

5
6

Hnh V.3.4.4.b. Cu trc ha hc ca IS4

Bng V.3.4.4. Cc s liu ph NMR ca IS4
C
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

C#

Ca, b

38,5

38,6

38,2

38,4

211,8
44,8

211,2
44,8

46,7
29,0
31,2
33,9
52,7
35,7
29,3
77,8
48,8
54,8
23,5
25,1

46,8
29,1
31,3
33,9
52,7
35,8
29,4
77,9
48,9
54,8
23,6
25,2

64,3
9,7
11,5
75,7
28,8
34,0

64,4
9,8
11,6
75,9
28,9
34,1

Ha, c
dng pic (J = Hz)
2,04 (1H, ddd, J = 13,0,6,5,4,5 Hz)
1,36 (1H, m)
2,42 (1H, dt, J = 3,5,6,5 Hz)
2,33 (1H, m)
2,20 (1H, t, J = 14,5 Hz)
2,10 (1H, dq, J = 14,5, 1 Hz)
1,53 (1H, m)
1,3-1,4 (2H, m)
1,72 (1H, m); 0,88 (1H, m)
1,38 (1H, m)
0,83 (1H, m)
1,76 (1H, m); 1,36 (1H, m)
3,37 (1H, dd, J = 11,0, 4,5 Hz)
1,00 (1H, m)
1,70 (1H, m); 1,22 (1H, m)
1,72 (1H, m)
1,50 (1H, m)
1,65 (1H, t, J = 6,5 Hz)
0,86 (3H, s)
1,02 (3H, s)
1,16 (3H, s)
1,55 (1H, m)
475

23
24
25
26
27
28
29

31,8

31,9

39,1
27,2
13,0
11,6

39,2
27,3
13,1
11,8

19,2
20,3

19,4
20,5

1,32 (1H, m)
1,82 (1H, brt, J = 10,5 Hz)
1,58 (1H, m)
0,68 (1H, q, J = 6,5 Hz)
0,18 (1H, m)
0,50 (1H, m)
0,18 (1H, m)
0,12 (1H, m)
1,03 (3H, d, J = 6,0 Hz)
0,94 (3H, d, J = 6,5 Hz)

o trong CDCl3, b 125MHz, c 500MHz, #s liu ca aragusterol B [280].

Hnh V.3.4.4.c. Ph 13C-NMR ca IS4

Phn tch chi tit cc tng tc trn ph cng hng t ht nhn hai chiu
HMQC, HMBC v COSY cho php xc nh v tr ca cc nhm chc cng nh
ton b cu trc phn t ca IS4. V tr ca nhm ktn ti C-3 c xc nh bi
cc tng tc HMBC t cc proton H-1, H-2 v H-3 sang C-3. Tng tc HMBC
t proton H-18 sang C-12 v H-21 sang C-20 cho php xc nh v tr tng ng
ca cc nhm hydroxyl ti C-12 v C-20 (Hnh V.3.4.4.g).

Hnh V.3.4.4.d. Ph HMQC ca IS4

476

Hnh V.3.4.4.e. Ph HMBC ca IS4

Hnh V.3.4.4.f. Ph COSY ca IS4

HO

HO

COSY
HMBC

Hnh V.3.4.4.g. Cc tng tc COSY v HMBC chnh ca IS4

477

T tt c cc phn tch nu trn, cng vi s ph hp hon ton v s liu

chuyn dch ha hc 13C-NMR ca IS4 so vi cc s liu c cng b
[280] cho php xc nh chnh xc cu trc ha hc ca IS4 l aragusterol B.
V.3.4.5. Hp cht IS5: iansterol C (cht mi)
Hp cht IS5 phn lp c di dng cht bt mu trng. Ph cng
hng t ht nhn 1H-NMR ca IS5 gn nh trng khp hon ton vi ph ca
IS4. im khc bit ln nht l s xut hin thm tn hiu ca 02 nhm metoxy
ti 3,14 (3H, s) v 3,19 (3H, s).
29

21

HO

18

12
19

11

13
14

H3CO

2
3

9
5

20
17
16

26

24

28
23

25
27

15

10

22

HO

7
6

OCH3

Hnh V.3.4.5.a. Cu trc ha hc ca IS5

Hnh V.3.4.5.b. Ph 1H-NMR ca IS5

478

Bng V.3.4.5. Cc s liu ph NMR ca IS5

Ca, b

C#

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26

34,9
28,3
100,3
35,4
42,4
28,3
31,3
33,8
52,4
35,7
29,3
77,8
48,9
54,5
23,7
26,8
48,2
8,3
11,5
65,7
50,5
72,2
40,2
34,9
27,6
12,5
12,4

35,18
28,60
100,60
35,61
42,60
28,62
31,59
34,07
52,95
35,94
29,31
78,21
48,87
55,07
23,68
25,29
64,40
9,81
11,76
75,89
28,93
34,15
31,94
39,26
27,31
13,12

18,9
19,1
47,8
47,8

19,40
20,45
47,69
47,73

27
28
29
OMe
OMe

11,78

Ha, c
dng pic (J = Hz)
1,58 (1H, m); 1,32 (1H,m)
1,90 (1H, m); 1,45 (1H, m)
1,63 (1H, m); 1,30 (1H, m)
1,31 (1H, m)
1,29 (1H, m); 1,18 (1H, m)
1,68 (1H, m); 0,88 (1H, m)
1,32 (1H, m)
0,83 (1H, m)
1,73 (1H, m); 1,27 (1H, m)
3,36 (1H, dd, J = 11,0, 4,5 Hz)
0,96 (1H, m)
1,68 (1H, m); 1,20 (1H, m)
1,71 (1H, m); 1,49 (1H, m)
1,65 (1H, overlapped)
0,84 (3H, s)
0,81 (3H, s)
1,16 (3H, s)
1,55 (1H, m); 1,32 (1H, overlapped)
1,58 (1H, m); 1,30 (1H, m)
0,66 (1H, quin-like, J = 6,5 Hz)
0,16 (1H, m)
0,50 (1H, m)
0,16 (1H, m)
0,11 (1H, m)
1,03 (3H, d, J = 6,0 Hz)
0,93 (3H, d, J = 6,5 Hz)
3,14 (3H, s)
3,19 (3H, s)

o trong CDCl3, b 125MHz, c 500MHz, #s liu ca aragusteroketal A [281].

Ph 13C-NMR ca IS5 hon ton ng t nh ca IS4 vi s xut hin cc

tn hiu ca 31 nguyn t cacbon. S xut hin thm 02 nhm metoxy cng c
khng nh trn ph 13C-NMR bi cc tn hiu metyl cng hng ti 47,69 v
47,73. Ngoi ra, tn hiu ca nhm ktn trong phn t ca IS4 c thay th
bi tn hiu ca mt cacbon bc 4 ni vi nguyn t xy ti 100.60 trong phn
t ca IS5. S chuyn dch mnh v vng trng thp ca tn hiu ny cho php
d on v tr ca n ti C-3 v n phi ni vi hai nguyn t xy.

479

Hnh V.3.4.5.c. Ph 13C-NMR ca IS5

Hnh V.3.4.5.d. Ph HMQC ca IS5

480

Hnh V.3.4.5.e. Ph HMBC ca IS5

Hnh V.3.4.5.f. Ph COSY ca IS5

481

HO

HO

COSY
HMBC

MeO
MeO

Hnh V.3.4.5.f. Cc tng tcHMBC v COSY chnh ca IS5

Hnh V.3.4.5.g. Ph khi lng FAB-MS ca IS5

Phn tch chi tit cc tng tc trn ph cng hng t ht nhn hai chiu
HMQC, HMBC v COSY cho php xc nh 2 nhm metoxy u gn vo C-3
cng nh ton b cu trc phn t ca IS5 (Hnh V.3.4.5.f). T tt c cc phn
tch nu trn, cng vi kt qu ph khi lng bn ph nguyn t nhanh FAB-MS
ti m/z 461 [M - CH2O + H]+ (tng ng vi cng thc C31H54O4, M = 490) cho
php xc nh nh c a ra trn hnh V.3.4.5.a. y l mt hp cht mi v
c chng ti t tn l iansterol C.

482

V.3.5. nh gi hot tnh gy c t bo ca cc hp cht

Tt c cc hp cht phn lp c t loi hi min Ianthella sp. c tin
hnh nh gi hot tnh gy c t bo. Kt qu th nghim cho thy, tt c cc
hp cht u th hin hot tnh mnh chng li 3 dng t bo ung th ngi th
nghim l MCF-7 (v), SK-Hep-1 (gan) v Hela (c t cung).
Bng V.3.5.1. Kt qu nh gi hot tnh gy c t bo ca cc hp cht
IC50 (M)
Hp cht
IS1
IS2
IS3
IS4
IS5
doxorubixin*

MCF-7
(v)
5,24
3,74
17,36
12,83
16,95
0,27

SK-Hep-1
(gan)
7,31
9,71
13,84
18,53
25,32
0,14

Hela
(c t cung)
15,63
12,84
18,67
27,80
24,61
0,19

*Cht chun dng

Kt qu ny hon ton ph hp vi cc nghin cu trc y. Nhiu cng

trnh cng b u khng nh rng cc hp cht sterol c cu trc b khung
tng t u th hin hot tnh gy c t bo rt mnh [280-282]. Cc tc nhn
gy c t bo c s dng trong iu tr mt s loi ung th nh ung th
phi, ung th v... v c coi l mt liu php chnh trong cc tr liu ung th
bng phng php ha hc. Cc tc nhn ny biu hin c tnh bng cch tiu
dit cc t bo ung th theo cc c ch khc nhau, ch yu l can thip vo qu
trnh phn chia t bo, c bit qu trnh sao chp DNA. Cc steroit c phn
lp t loi hi min Ianthella sp. c cu trc c o v th hin hot tnh gy
c t bo cao i vi cc dng t bo ung th c th nghim, iu ny gi
m cho cc nghin cu tip theo theo hng ng dng lm dc liu.

483

tng hp cc cht phn lp c t loi hi min

Ianthella sp.
29

29
21

22

18

19

12
11

13
14

2
3

HO

10
5

21

26

24

28

20

12

25

23

11

19

13

27

17
16
15

22

18

14
1

2
3

28

20

27

17
16
15

IS1 - Petrosterol

IS2 - Iansterol A (cht mi)

29

29

21
21

22

18
12
19

11

13
14

2
3

HO

25

23

10

26

24

20

26

24

28

HO

25

23

27

17

14
2

13

15

10

18

12
11

19

16

9
8

10

IS4 - aragusterol B

IS3 - Iansterol B (cht mi)

29
21

HO

HO
18

12
19

11

13
14

H3CO

2
3

9
5

20

26

24

28
23

25
27

17
16
15

10

22

7
6

OCH3

IS5 - Iansterol C (cht mi)

484

22

HO
20

28
23

25
27

17
16
15

26

24

chng VI: Tng hp cc hp cht phn lp

c trong khun kh ti KC09.09/06-10 v hot
tnh sinh hc ca chng
Trong khun kh ti KC09.09/06-10, chng ti tin hnh phn lp v
xc nh cu trc c 51 hp cht. Trong 18 hp cht c cu trc mi ln u
tin c phn lp t thin nhin. Cc hp cht phn lp c c tin hnh
nh gi hot tnh: gy c t bo, chng xy ha v khng vi sinh vt kim nh.
Ngoi ra, thng qua hp tc quc t, hot tnh chng long xng thng qua cc c
ch kch thch s bit ha ca nguyn bo xng v khng yu t phin m NF-B
ca mt s hp cht cng c tin hnh nh gi. Ton b cc cht c phn
lp, cu trc v hot tnh sinh hc ca chng c tng hp di y.
VI.1. Lp cht ditecpen dng cembranoit
Cembranoit l cc hp cht ditecpen c cu trc vng ln 14 cacbon trong
phn t. Khung cembranoit thng c bit n vi hot ng sinh hc rt a
dng. Cc loi san h mm c bit n nh l ngun sn sinh cc cembranoit di
do, trong chi Sarcophyton thng cha hm lng cao cc hp cht ny (i khi
ln ti 10% hm lng kh). Cc cembranoit ng vai tr quan trng cho s sinh
tn ca cc loi san h ny, chng hot ng nh h thng phng v, cnh tranh sn
mi, v ti sinh. Trong khun kh ti, chng ti phn lp v xc nh c cu
trc ca 07 hp cht cembranoit t loi san h mm Sarcophyton mililatensis, v
mt iu rt ng quan tm l c ti 05 hp cht c cu trc mi, ln u tin
c phn lp t thin nhin. Di y l cc cembranoit c phn lp, xc nh
cu trc v nh gi HTSH.
1. Hp cht (-)-7-hydroxy-8-methoxydeepoxysarcophytoxide (Cht mi, ln
u tin c phn lp t thin nhin)
- Cu trc ha hc:

18

HO

- c phn lp t loi: Sarcophyton

mililatensis.
- Hot tnh sinh hc:
+Hot tnh chng long xng

16

21
19

OMe
1

15

17

14

11

M
0,3
3
30

20

Hot lc ca
enzym ALP
(%)
127,81,392
125,12,760
115,56,288

Hm lng
collagen
(%)
108,21,860
101,92,680
91,483,231

S khong ha
(%)
110,62,470
106,70,403
105,12,619

- Cht du khng mu
- Cng thc phn t: C21H34O3, M = 334
+ Hot tnh gy c t bo
- Ph khi lng: m/z 357 [M+Na]+ 317 [MDng KB: IC50 = 3,23 g/ml
H2O+H]+, 303 [M-MeOH+H]+, 285 [MDng FL: IC50 = 2,77 g/ml
MeOH-H2O+H]+
Dng Hep-G2: IC50 = 3,45 g/ml
- c cng b trong cng trnh: Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van
Kiem, Chau Van Minh, Eun Mi Choi, and Young Ho Kim, New Cembranoid Diterpenes
from the Vietnamese Soft Coral Sarcophyton mililatensis Stimulate Osteoblastic
Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin, 2008, 56(7),
988-992.
485

2. Hp cht (-)-17-hydroxysarcophytonin A (Cht mi, ln u tin c phn

lp t thin nhin)
- Cu trc ha hc:
- Hot tnh sinh hc:
+Hot tnh chng long xng

H
O

OH

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

131,06,834
127,21,292
130,83,774

101,03,154
102,51,832
111,41,208

97,111,485
101,72,556
107,93,767

+ Hot tnh gy c t bo
Dng FL: IC50 = 3,06 g/ml
Dng Hep-G2: IC50 = 3,69 g/ml

- Cht du khng mu
- Cng thc phn t: C20H30O2, M = 302
- Ph khi lng: m/z 325 [M+Na]+, 303
[M+H]+, 285 [M-H2O+H]+
- c phn lp t loi: Sarcophyton
mililatensis.
- c cng b trong cng trnh: Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van
Kiem, Chau Van Minh, Eun Mi Choi, and Young Ho Kim, New Cembranoid Diterpenes
from the Vietnamese Soft Coral Sarcophyton mililatensis Stimulate Osteoblastic
Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin, 2008,
56(7), 988-992.

3. Hp cht sarcophytol V (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
- c phn lp t loi: Sarcophyton
mililatensis.
- Hot tnh sinh hc:
OH +Hot tnh chng long xng

OH

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

127,02,449
121,02,237
113,22,973

102,70,137
101,01,938
110,41,440

96,771,650
97,721,546
101,60,990

+ Hot tnh gy c t bo
- Cht du khng mu
Dng KB: IC50 = 3,77 g/ml
- Cng thc phn t: C20H32O2, M = 304
- Ph khi lng: m/z 327 [M+Na]+, 287
[M-H2O+H]+, 269 [M-2H2O+H]+
- c cng b trong cng trnh: Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van
Kiem, Chau Van Minh, Eun Mi Choi, and Young Ho Kim, New Cembranoid Diterpenes
from the Vietnamese Soft Coral Sarcophyton mililatensis Stimulate Osteoblastic
Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin, 2008,
56(7), 988-992.

486

4. Hp cht sarcophytol X (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
- c phn lp t loi: Sarcophyton
mililatensis.
- Hot tnh sinh hc:
Chng xy ha trn h DDPH vi gi tr
ED50 = 75,5 g/ml

O
OH
O

OH
OH

- Cht du khng mu
- Cng thc phn t: C20H32O5, M = 352
- Ph khi lng: m/z 334,8 [M-H2O+H]+,
316,8 [M-2H2O+H]+, 298,8 [M-3H2O+H]+)
- Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van Kiem, and Chau Van Minh,
Sarcophytol X, a new cembranoid diterpene from the vietnamese sea soft coral
Sarcophytol mililatensis, Tp ch Khoa hc (Trng i hc S phm H Ni 2), 2007,
tp 1, 55-60.

5. Hp cht (+)-7,8-dihydroxydeepoxysarcophytoxide (Cht mi, ln u

tin c phn lp t thin nhin)
- Cu trc ha hc:

H
O

HO
HO

- c phn lp t loi: Sarcophyton

mililatensis.
- Hot tnh sinh hc:
+Hot tnh chng long xng
M

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

128,92,660
139,52,237
123,41,814

109,51,898
106,84,880
99,794,012

100,71,684
104,01,277
105,42,868

+ Hot tnh gy c t bo
Dng KB: IC50 = 4,23 g/ml
Dng FL: IC50 = 1,97 g/ml
Dng Hep-G2: IC50 = 3,95 g/ml

- Cht du khng mu
- Cng thc phn t: C20H32O3, M = 320
- Ph khi lng: m/z 343 [M+Na]+, 303
[M-H2O+H]+
- c cng b trong cng trnh: Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van
Kiem, Chau Van Minh, Eun Mi Choi, and Young Ho Kim, New Cembranoid Diterpenes
from the Vietnamese Soft Coral Sarcophyton mililatensis Stimulate Osteoblastic
Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin, 2008,
56(7), 988-992.

487

6. Hp cht (+)-sarcophine
- Cu trc ha hc:
- Hot tnh sinh hc:
+Hot tnh chng long xng

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

133,32,091
130,26,923
121,91,062

103,50,836
109,72,000
104,01,716

96,171,540
99,870,941
99,661,515

+ Hot tnh gy c t bo
Dng FL: IC50 = 4,21 g/ml
Dng Hep-G2: IC50 = 3,55 g/ml
+ Hot tnh chng xy ha
ED50 = 62,3 g/ml

- Cht du khng mu
- Cng thc phn t: C20H32O3, M = 320
- Ph khi lng: m/z 339 [M+Na]+, 317
[M+H]+, 299 [M-H2O+H]+ .
- c phn lp t loi: Sarcophyton
mililatensis.
- c cng b trong cng trnh: Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van
Kiem, Chau Van Minh, Eun Mi Choi, and Young Ho Kim, New Cembranoid Diterpenes
from the Vietnamese Soft Coral Sarcophyton mililatensis Stimulate Osteoblastic
Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin, 2008,
56(7), 988-992.

7. Hp cht sarcophytoxide
- Cu trc ha hc:

H
O

- Hot tnh sinh hc:

+Hot tnh chng long xng

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

127,82,995
125,13,296
117,42,995

103,23,421
106,91,636
109,11,734

96,102,443
97,783,635
102,81,296

+ Hot tnh gy c t bo
Dng FL: IC50 = 4,32 g/ml
- Cht du khng mu
- Cng thc phn t: C20H30O2, M = 302
- Ph khi lng: m/z 325 [M+Na]+, 303
[M+H]+, 285 [M-H2O+H]+.
- c phn lp t loi: Sarcophyton
mililatensis.
- c cng b trong cng trnh: Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van
Kiem, Chau Van Minh, Eun Mi Choi, and Young Ho Kim, New Cembranoid Diterpenes
from the Vietnamese Soft Coral Sarcophyton mililatensis Stimulate Osteoblastic
Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin, 2008, 56(7),
988-992.

488

VI.2. Lp cht 9,11-secosterol

9,11-Secosteroit l cc steroit c cu trc vng C b m vng ti v tr C-9.
Tt c cc hp cht 9,11-secosterol c phn lp u c nhm kton C-9.
Chng thng c nhm aldehit (CHO) hay hydroxyl (OH) v tr C-11, cc nhm
OH vng A v B, v i khi c cu epoxit v tr C-5 v C-6. Cc hp cht 9,11secosteroit thng c ni i v tr C-5/C-6, C-7/C-8 hay c cu trc vng no.
Chng thng c phn lp t hi min, rut khoang v t hi tiu A. conicum.
Nghnh rut khoang li c chia thnh cc lp Hydrozoa (thy tc), Cubozoa,
Scyphozoa (sa) v Anthozoa (san h). Lp Anthozoa li c chia thnh 2 lp ph
l Octocorallia v Hexacorallia. Lp ph Octocorallia li c chia thnh 5 b,
trong cc b Gorgonacea (san h sng), Alcyonacea (san h mm) v Stolonifera
l ngun sn sinh di do cc hot cht 9,11-secosteroit.
Mc d cho ti nay, mt s lng ln cc hp cht steroit c phn lp
v xc nh cu trc, ch c mt s lng t cc hp cht secosterol c cng b.
Chng thng th hin hot tnh gy c t bo v chng tng sinh rt nhiu dng
t bo ung th khc nhau nh C6 (t bo lioma chut), J774 (bch cu n nhn to
v i thc bo chut), LnCap (t bo ung th tuyn tin lit), Calu-3 (ung th
phi).... Trong khun kh ti, chng ti phn lp c 03 hp cht 9,11scosteroit t cc loi san h mm sinh sng ti cc vng bin Vit Nam. Mt iu
th v l c ti 02 trong s 03 hp cht phn lp c c cu trc mi, ln u
tin c phn lp t thin nhin. Di y l cc hp cht phn lp c.
8. Hp cht cladiasterol (Cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:
HO

O H
O

OH
H

HO

O
HO

OH

- Ph khi lng phn gii cao:

m/z 615,389 [M+Na]+ (tnh
ton l thuyt cho
C34H56O8Na: 615,3873)
- c phn lp t loi:
Cladiella sp.
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng KB: IC50 = 4,43 g/ml
+ Hot tnh chng xy ha
ED50 = 77,8 g/ml

- Cht bt mu trng
- Cng thc phn t: C34H56O8, M = 592
- Ph khi lng: m/z 615.2 [M+Na]+, 454.2 [Mfucose+Na]+, 591.4 [M-H]- c cng b trong cng trnh: Chau Van Minh, Phan Van Kiem, and Nguyen Xuan
Cuong, New 9,11-secosterol glycoside and ceramide from a soft coral Cladiella sp.
living in Vietnamese sea, Asean Journal on Sicience and Technology for Development,
2007, 24(3), 275-283.

489

9. Hp cht sarcomilasterol (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
28

21

26
24

17

11

HOH2C

27

13

19

25

20

18

- c phn lp t loi: Sarcophyton

mililatensis.
- Hot tnh sinh hc:
+Hot tnh chng long xng

14

1
8

10
3

5
6

HO

Hot lc ca
enzym ALP
(%)

Hm lng
collagen (%)

S khong ha
(%)

0,3
3
30

119,15,214
118,01,486
86,410,212

106,51,822
104,01,254
69,10,495

101,11,8564
107,21,680
89,383,609

+ Hot tnh gy c t bo
Dng KB: IC50 = 2,23 g/ml
Dng FL: IC50 = 3,57 g/ml
Dng Hep-G2: IC50 = 2,65 g/ml

OH

- Tinh th hnh kim mu trng: mp 160161oC

- Cng thc phn t: C28H48O4, M = 448
- Ph khi lng: m/z 471,1 [M+Na]+,
449,1 [M+H]+, 431,1 [M-H2O+H]+
- c cng b trong cng trnh: Chau Van Minh, Nguyen Xuan Cuong, Tran Anh
Tuan, Eun Mi Choi, Young Ho Kim, and Phan Van Kiem, A new 9,11-secosterol from
the Vietnamese Sea soft coral Sarcophyton mililatensis, increases the function of
osteoblastic MC3T3-E1 cells, Natural Product Comunications (USA), 2007, 2(11),
1095-1100.

10. Hp cht 3,6,11-trihydroxy-24-methylene-9,11-secocholest-7-en-9-one

- Cu trc ha hc:

28
26

21

HO

20

11
12
19

13

HO

10

23

24

25

27

17
16

14
9

22

18

15

- Hot tnh sinh hc:

+ Hot tnh khng NF-B:
IC50 = 32,5 1,80 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 4,23 g/ml
Dng FL: IC50 = 2,97 g/ml
Dng Hep-G2: IC50 = 1,95
g/ml

7
6

H
OH

- Tinh th hnh kim mu trng: mp 162-164oC

- Cng thc phn t: C28H46O4, M = 446
- Ph khi lng: m/z 447,2 [M + H]+, 445,4 [M-H]- c phn lp t loi: Cladiella sp.
- c cng b trong cng trnh: Chau Van Minh, Phan Van Kiem, and Nguyen Xuan
Cuong, New 9,11-secosterol glycoside and ceramide from a soft coral Cladiella sp.
living in Vietnamese sea, Asean Journal on Sicience and Technology for Development,
2007, 24(3), 275-283.

490

VI.3. lp cht tritecpen saponin

Holothurin l cc hp cht triterpen glycosit, hu ht thuc khung holostane
[(20S)-20-hydorxy-5-lanostane-18-carboxylic acid (18-20) lactone]. Cc hp cht
ny thng c ph hot tnh rng nh: gy c t bo, khng nm, tan mu, iu
bin min dch... c trn 100 hp cht thuc nhm cht ny c phn lp t
cc loi hi sm. Hu ht cc hp cht ny c cu trc khung c bn holostane gn
vi mt chui cc n v ng (2-6 n v ng) v tr C-3. Cc hp cht ny
thng c nhiu hot tnh qu bu nn c gi tr rt cao. n c nh hp ct
holothurin A c hng Sigma rao bn vi gi 5000 USD/1g.
Trong khun kh ti, t loi hi sm Holothuria scabra, chng ti phn
lp v xc nh cu trc c 06 hp cht holothurin trong c 03 hp cht mi.
iu ny gp phn khng nh gi tr thc t ca ngun dc liu bin ng thi t
c s khoa hc cho cc nghin cu tip theo v cc loi hi sm ca Vit Nam nhm
pht trin cc dc cht mi t ngun dc liu qu bu v di do ca t nc.
11. Hp cht holothurigenin B (Cht mi, ln u tin c phn lp t thin
nhin)
- Cu trc ha hc:
23

24

21

25
20

18
19

10

13
9

O O

OH

15

6
1'

OH
6'' CH3

28

29

2'

27

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 2,33
g/ml
+ Hot tnh khng VSV kim nh
Khng cc chng nm:
Aspergillus niger, Fusarium
oxysporum, Candida albicans,
v Saccharomyces cerevisiae
vi gi tr MIC = 50 g/ml

OH

OH

5'

4''

30

4'

17

12

11

26
22

1''

OH
2''

OH

- Cht bt v nh hnh mu trng

- Cng thc phn t: C41H62O13 (M = 762)
- Ph khi lng: m/z 762,7 [M+H]+, 785.3
[M+Na]+, 761.3 [M-H]- c phn lp t loi: Holothuria scabra.
- c cng b trong cng trnh: Nguyen Van Thanh, Nguyen Hai Dang, Phan Van
Kiem, Nguyen Xuan Cuong, Hoang Thanh Huong, and Chau Van Minh. A new
triterpene glycoside from the sea cucumber Holothuria scabra collected in Vietnam.
Asean Journal on Science and Technology for Development, 2006, 23(4), 253-259.

491

12. Hp cht holothurin A3 (Cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
- Cht bt v nh hnh mu
trng
- Cng thc phn t:
O

21

HO

17

12

26

23

25

OH

13

27

10

15
30

3
4

5'

O O

Xyl
4'

20

18

11

19

24

22

OH

NaO3SO

29

28

1'
2'

6'' CH3
O O

Holothuria scabra.

1''

OH

C54H85O28NaS, M = 1236
- Ph khi lng: m/z 1259,5
[M+Na]+, 1235,5 [M-H]- Ph khi lng phn gii cao:
m/z 1237,5027 [M+H]+ (tnh ton
l thuyt cho C54H86O28NaS:
1237,4924)

- c phn lp t loi:

Qui
4''
6'''

OH

CH2OH

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
MeGlc
Dng FL: IC50 = 0,39 g/ml
Dng Hep-G2: IC50 = 0,23
g/ml
Dng KB: IC50 = 0,57 g/ml
- c cng b trong cng trnh: Nguyen Hai Dang, Nguyen Van Thanh, Phan Van
Kiem, Le Mai Huong, Chau Van Minh and Young Ho Kim. Two new triterpene
glycosides from the Vietnamese sea cucumber Holothuria scabra. Archives of
Pharmacal Research, 2007, 30(11), 1387-1391.
- Chu Vn Minh, Phan Vn Kim, Nguyn Hi ng, Nguyn Vn Thanh, L Mai
Hng, Cng Thung, Trn Thu Hng, Nguyn Xun Cng. Hp cht Holothurin
A3 c hot tnh chng ung th v phng php chit hp cht ny t loi hi sm
Holothuria scabra, Bng c quyn sng ch s 6852, cp theo quyt nh s 2394/QSHTT, ngy 14 thng 02 nm 2008 ca Cc S hu Tr Tu Vit Nam
O O

6''''

CH2OH

Glc

4'''

OH

1'''

OH

4''''

2'''

1''''

OCH3

OH

2''

OH

2''''

OH

13. Hp cht holothurin A4 (Cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
- Cht bt v nh hnh mu
trng
- Cng thc phn t:
OH

21

22

HO

19

10

17

13

OH

27

15
30

O O

OH

1'

NaO3SO

6
28

Holothuria scabra.

2'

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 0,57
g/ml
Dng KB: IC50 = 1,12 g/ml

O O
4''
6'''

CH2OH
1''''

1'''

OCH3
OH

2''''

OH

2''

OH

4'''

4''''

1''

OH

CH2OH
O O

OH

C54H87O27SNa (M=1222)
- Ph khi lng: m/z 1245,2
[M+Na]+, 1221,4 [M-H]-

- c phn lp t loi:

29

6'' CH3

6''''

26

25

23

5'

4'

20

18

12

11

24

2'''

OH

- c cng b trong cng trnh: Nguyen Hai Dang, Nguyen Van Thanh, Phan Van
Kiem, Le Mai Huong, Chau Van Minh and Young Ho Kim. Two new triterpene
glycosides from the Vietnamese sea cucumber Holothuria scabra. Archives of
Pharmacal Research, 2007, 30(11), 1387-1391.

492

14. Hp cht holothurin A

- Cu trc ha hc:
21

O
HO

10

Xyl

- Ph khi lng: m/z 1243,3

[M+Na]+
- c phn lp t loi:
Holothuria scabra, H.
martensii v H. vagabunda.
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 4,08
g/ml
Dng KB: IC50 = 1,28 g/ml
Dng FL: IC50 = 1,54 g/ml

27

OH

15
30

O O

OH

6
1'

NaO3SO
6'' CH3

29

28

2'

Qui
4''

26

25

5'

O O
1''

OH

6'''

20
17

13

4'

22

18
12

11

19

24

23

CH2OH
OH

CH2OH
O

1'''
2'''

OH

MeGlc
4''''

Glc

4'''

6''''

OH

1''''

OCH3

OH

2''

O O

2''''

OH

- Cht bt v nh hnh mu trng: mp 228-230oC

- Cng thc phn t: C54H85O27SNa (M=1220)
- c cng b trong cng trnh: Nguyen Van Thanh, Nguyen Hai Dang, Phan Van
Kiem, Nguyen Xuan Cuong, Hoang Thanh Huong, and Chau Van Minh. A new
triterpene glycoside from the sea cucumber Holothuria scabra collected in Vietnam.
Asean Journal on Science and Technology for Development, 2006, 23(4), 253-259.

15. Hp cht holothurin A2

- Cu trc ha hc:
21
22

O
HO

19

10

17

27

9
15
30

O O

OH

6
1'

NaO3SO

28

29

2'

6'' CH3
O O
4''
6'''

1''

OH

CH2OH
O O

6''''

CH2OH

4''''

OH

1''''

OCH3

- c phn lp t loi:
Holothuria scabra.
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng KB: IC50 = 2,42
g/ml
Dng FL: IC50 = 1,00
g/ml
Dng Hep-G2: IC50 = 1,06
g/ml

2''

OH

4'''

25

23

5'

26

OH

13

4'

20

18
12

11

24

1'''

OH

2'''

OH
2''''

OH

- Cht bt v nh hnh mu trng: mp 246-2480 C

- Cng thc phn t: C54H87O26SNa, M = 1206
- Ph khi lng: m/z 1229,6 [M+Na]+
- c cng b trong cng trnh: Nguyen Van Thanh, Nguyen Hai Dang, Phan Van
Kiem, Nguyen Xuan Cuong, Hoang Thanh Huong, and Chau Van Minh. A new
triterpene glycoside from the sea cucumber Holothuria scabra collected in Vietnam.
Asean Journal on Science and Technology for Development, 2006, 23(4), 253-259.

493

16. Hp cht holothurin B

- Cu trc ha hc:
21

O
HO

19

10

Xyl

13

OH

15
30

O O

OH

Qui

6
1'

NaO3SO
6'' CH3

OH

17

5'

4''

22
20

12

11

4'

O
18

24

23

28

29

2'
O O

26

25
27

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng KB: IC50 = 2,85 g/ml
Dng FL: IC50 = 2,14 g/ml
Dng Hep-G2: IC50 = 1,99
g/ml
+ Hot tnh khng VSV kim nh
Khng cc chng nm:
Aspergillus niger, Fusarium
oxysporum, Candida albicans,
v Saccharomyces cerevisiae
vi gi tr MIC = 50 g/ml

1''

OH
2''

OH

- Cht bt v nh hnh mu trng: mp 251-253oC

- Cng thc phn t: C41H63O17NaS (M = 882)
- Ph khi lng: m/z 905 [M+Na]+
- c phn lp t loi: Holothuria scabra, H.
martensii v H. vagabunda.
- c cng b trong cng trnh: Nguyen Van Thanh, Nguyen Hai Dang, Phan Van
Kiem, Nguyen Xuan Cuong, Hoang Thanh Huong, and Chau Van Minh. A new
triterpene glycoside from the sea cucumber Holothuria scabra collected in Vietnam.
Asean Journal on Science and Technology for Development, 2006, 23(4), 253-259.

VI.4. Lp cht cerebrosit v ceramit

Cerebrosit, mt h glycosphingolipit, l thnh phn quan trng ca rt nhiu
m v c quan khc nhau trong c th sinh vt. V mt ha hc, chng c to
thnh bi mt ng hexose v mt n v ceramit thng bao gm mt
aminoalcohol mch di gi l baz sphingoid (sphingosine hay sphingol) v mt
axit bo mch di lin kt vi nhm amit. Cu trc tng th ca mt h cerebrosit
thin nhin u tin c Carter v cng s pht hin vo nm 1950. Sau ,
Shapiro v Flowers cng ln u tin v c lp cng b quy trnh tng hp ton
phn ca mt cerebrosit vo nm 1961.
V phng din sinh hc, mt s cerebrosit c chng minh gi vai tr
h tr cu trc v xc nh kt cu ca cc mng t bo, v hot ng nh cc cht
iu tit ca cc qu trnh sinh hc nh qu trnh hot ha, qu trnh dnh kt t bo,
trao i thng tin ni bo v s pht trin ca t bo. Hn na, cc cerebrosit trong
mng t bo gi vai tr sinh hc quan trng l cc khng nguyn b mt v cc th
th. c bit, ngy cng c nhiu bng chng ch ra rng cc cerebroside c cc
chc nng sinh hc rt a dng, hu ht cc chc nng ny u lin quan n c
tnh lng cc ca phn t. Chng hn nh, nhn t pht trin thn kinh (NGF Nerve growth factor), mt i phn t sinh hc l nhn t u tin v c xc nh
r nht, iu tit s pht trin, bit ha v s sng st ca cc nron v n c coi
494

l mt ng vin cho cc thuc tr bnh Alzheimer (bnh qun). Tuy nhin, phn t
NGF qu ln v mt kch thc v qu a nc i qua cc vch ngn mu no, v
khng c con ng an ton no khc a cc NGF vo trong no tht. Do ,
cc cht phn t lng thp c tc dng tng t cc NGF nh cc cerebrosit l cc
ng vin y ha hn cho vic tr bnh Alzheimer. 01 ceramit mi v 01 cerebrosit
mi trong tng s 03 hp cht c phn lp trong khun kh ti l:
17. Hp cht cladiaceramide (Cht mi, ln u tin c phn lp t thin
nhin)
- Cu trc ha hc:
OH
O

1'

3'

16'

10

HN
18

HO

OH

- c phn lp t loi: Cladiella

sp.
- Ph khi lng phn gii cao:
m/z 574,489 [M+Na]+ (tnh
ton l thuyt cho
C34H65NO4Na: 574,4811)
- Hot tnh sinh hc:
+ hot tnh khng NF-B
IC50 = 14,8 1,73 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 4,43 g/ml

- Cht bt v nh hnh mu trng

- Cng thc phn t: C34H65NO4 (M = 551)
- Ph khi lng: m/z 574,4 [M+Na]+, 552,4
[M+H]+, 534,4 [M-H2O+H]+, 550,4 [M-H]- c cng b trong cng trnh: Chau Van Minh, Phan Van Kiem, and Nguyen Xuan
Cuong, New 9,11-secosterol glycoside and ceramide from a soft coral Cladiella sp.
living in Vietnamese sea, Asean Journal on Sicience and Technology for Development,
2007, 24(3), 275-283.

18. Hp cht archastercerebroside (cht mi, ln u tin c phn lp t

thin nhin)
- Cu trc ha hc:
OH
OH OH
6"

4"

H
HO

(CH2)21CH3

H O

5"

3"

2'

1'

Gal

2"

OH
H

OH

NH

1"

(CH2)9CH(CH3)2

OH

- Ph khi lng phn gii cao:

m/z 840,6734 [M+Na]+ (tnh
ton l thuyt cho
C46H91NO10Na: 840,6541)
- Hot tnh sinh hc:
+ hot tnh khng NF-B
IC50 = 34,7 1,84 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 4,45 g/ml
Dng FL: IC50 = 3,91 g/ml
Dng Hep-G2: IC50 = 2,35
g/ml

- Cht bt v nh hnh mu trng: mp 221-222oC

- Cng thc phn t: C46H91NO10 (M = 817)
- Ph khi lng: m/z 818,8 [M+H]+, 816,8 [M-H]- c phn lp t loi: Archaster typicus
- c cng b trong cng trnh: Chau Van Minh, Nguyen Van Thanh, Nguyen Xuan
Cuong, Phan Van Kiem, Nunziatina De Tommasi and Lam Ngoc Thiem, A new
galactocerebroside from the starfish Archaster typicus, Tuyn tp cc cng trnh Hi
ngh Khoa hc v Cng ngh Ha hc Hu c ton quc ln th IV, H Ni, 10-2007, tr
446-450.

495

19. Hp cht cladiacerebroside (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:

- Ph khi lng: m/z: 848,6

[M+H]+, 846,8 [M-H]O
- c phn lp t loi:
16
Cladiella sp.
HN
OH
O
- Hot tnh sinh hc:
O
HO
+ Hot tnh gy c t bo
7
HO
SO3H
Dng KB: IC50 = 3,77 g/ml
HO
Dng Hep-G2: IC50 = 4,72
- Cht bt v nh hnh mu trng
g/ml
- Cng thc phn t: C45H85NO11S, M = 847
+ Hot tnh chng xy ha
ED50 = 65,5 g/ml
- c cng b trong cng trnh: Chau Van Minh, Nguyen Hai Dang, Nguyen Xuan
Cuong, Phan Van Kiem, Hoang Thanh Huong, Bioacive constituents from marine
organisms inhabiting in Vietnames sea. Journal of Science and Technology (Vietnam),
2007, 45(6-DB), 1-18.
OH

1'

20'

6"

4"

18

5"

1"

2"

3"

19

VI.5. Lp cht axit bo khng no b brm ha

y l cc axit bo a ni i, ni ba c cha t 1 n nhiu nguyn t Brm
trong phn t. Theo ti liu cng b, cc hp cht axit mch di a ni i v ni
ba b brm ha thng c hot tnh khng sinh v gy c t bo. c bit, mt s
hp cht th hin hot tnh c ch enzym HIV protease, mt enzym thit yu cho s
ti bn ca HIV. Ngoi ra, chng cn c hot tnh km hm tc nhn i khng lin
kt vi th th Adenosine. Khi th th ny c hot ha s mang n cc hiu qu
tch cc nh gim nhp tim, bo v no v tiu m. Trong khun kh ti, chng
ti phn lp c 02 hp cht thuc nhm axit bo a ni i, ni ba b brm ha
v mt iu th v l chng u c cu trc mi, ln u tin c phn lp t
thin nhin. Di y l cc hp cht c phn lp v xc nh cu trc.
20. Hp cht axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17triynoic (cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:

6
7
9

COOH
1

17
15

11

Br
19

- Cht kt tinh mu trng

- Cng thc phn t: C20H21BrO2, M = 372
- Ph khi lng: m/z 394.8/396.8 [M+Na]+ vi t l 1/0,98 v cng tn hiu
- c phn lp t loi: Xestospongia testudinaria.
- Hot tnh khng vi sinh vt kim nh: khng mnh ba chng vi sinh vt kim nh l:
trc khun m xanh Pseudomonas aeruginosa, khng t cu vng Staphylococcus aureus
v khng nm si Fusarium oxysporum vi gi tr MIC thp l 50 g/ml.
- c cng b trong cng trnh:
496

1.Chu Vn Minh, Phan Vn Kim Trn Anh Tun v Nguyn Xun Cng, Two new
brominated fatty acids from the sponge Xestospongia testudinaria living in Vietnamese
sea, Advance in Natural Science, 2008, 9(2), 167-171
2. Chu Vn Minh, Nguyn Xun Cng, Phan Vn Kim, L Mai Hng, Cng Thung
Hp cht axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic v phng
php chit hp cht ny t loi hi min Xestospongia testudinaria, Sng ch np ti
Cc s hu tr tu Vit Nam, s n 1-2007-01616 SC ngy 8/8/2007 (ang thm nh
ni dung).

21. Hp cht axit 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19-triynoic

(cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:

8
9

COOH

11

19
13

17

22

Br

21

- Cht kt tinh mu trng

- Cng thc phn t: C22H27O2Br, M = 402
- Ph khi lng: m/z 424.9/426.9 [M+Na]+ vi t l 1/0,98 v cng tn hiu
- c phn lp t loi: Xestospongia testudinaria.
- Hot tnh khng vi sinh vt kim nh: khng mnh ba chng vi sinh vt kim nh l:
trc khun m xanh Pseudomonas aeruginosa, khng t cu vng Staphylococcus aureus
v khng nm si Fusarium oxysporum vi gi tr MIC thp l 50 g/ml.
- c cng b trong cng trnh: Chu Vn Minh, Phan Vn Kim, Trn Anh Tun v
Nguyn Xun Cng, Two new brominated fatty acids from the sponge Xestospongia
testudinaria living in Vietnamese sea, Advance in Natural Science, 2008, 9(2), 167-171

VI.6. lp cht steroit c cu trc vng propan mch

nhnh
22. Hp cht iansterol A (cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:

29
21

22

18
12
19

11

13
14

2
3

26

24

28
23

25

17
16

27

15

10
5

20

7
6

- Cht bt mu trng
- Cng thc phn t: C29H44O2 (M = 424)
- Ph khi lng: m/z 425 [M+H]+

497

- Ph khi lng phn gii cao HRFAB-MS: m/z 425,3397 [M+H]+

(tnh ton l thuyt cho C29H45O2
l 425,3375)
- c phn lp t loi: Ianthella
sp.
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 3,74 M
Dng SK-Hep-1: IC50 = 9,71 M
Dng Hela: IC50 = 12,84 M

23. Hp cht iansterol B (cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:

29
21

22

18

11

19
1

HO

25

23

27

17

13

16

14
2

28

20

12

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 17,36 M
Dng SK-Hep-1: IC50 = 13,84 M
Dng Hela: IC50 = 18,67 M

26

24

15

9
8

10

- Cht bt mu trng
- Cng thc phn t: C29H48O2 (M = 428)
- Ph khi lng: m/z 429 [M+H]+
- Ph khi lng phn gii cao HR-EI-MS: m/z
428,3656[M+H]+ (tnh ton l thuyt cho
C29H48O2 l 428,3654)
- c phn lp t loi: Ianthella sp.

24. Hp cht iansterol C (cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:

29

21

HO

18

12
19

11

13
14

H3CO

2
3

9
5

20

26

24

28
23

25
27

17
16
15

10

22

HO

7
6

OCH3

- Cht bt mu trng
- Cng thc phn t: C31H54O4, M = 490
- Ph khi lng: m/z 461 [M - CH2O + H]+
- c phn lp t loi: Ianthella sp.

498

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 16,95 M
Dng SK-Hep-1: IC50 = 25,32 M
Dng Hela: IC50 = 24,61 M

25. Hp cht petrosterol

- Cu trc ha hc:

29
21

22

18
12

19
1

2
3

HO

11
9

10
5

28

20

13

17
16

14

15

26

24
25

23

27

- c phn lp t loi: Ianthella sp.

- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 5,24 M
Dng SK-Hep-1: IC50 = 7,31 M
Dng Hela: IC50 = 15,63 M

8
7

- Cht bt mu trng
- Cng thc phn t: C29H48O, M = 412

26. Hp cht aragusterol B

- Cu trc ha hc:

29

21

HO

22

HO
18

11

2
3

27

16
15

- c phn lp t loi: Ianthella sp.

- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 12,83 M
Dng SK-Hep-1: IC50 = 18,53 M
Dng Hela: IC50 = 27,80 M

10

25

23

17

13
14

24

28

20

12
19

26

- Cht bt mu trng
- Cng thc phn t: C29H48O3, M = 444

VI.7. Lp cht steroit mang nhiu nhm hydroxyl trong

phn t (polyhydroxylated steroids)
27. Hp cht 27-norcholestan-3,4,5,6,8,14,15,24R-octol
- Cu trc ha hc:
OH
21

22
20

18
12
11

19

HO

10

25

23

17

13

OH

24

26

16
14

15

8
7

OH

OH

OH
OH

OH

- Ph khi lng: m/z 509,2

[M+Na]+, 485,4 [M-H]- c phn lp t loi: Archaster
typicus
- Hot tnh sinh hc:
+ Hot tnh khng NF-B:
IC50 = 30,82,12 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 3,37 g/ml
Dng Hep-G2: IC50 = 2,69 g/ml

- Tinh th hnh kim mu trng: mp 294oC

- Cng thc phn t: C26H46O8 (M = 486)
- c cng b trong cng trnh: Chau Van Minh, Nguyen Xuan Cuong, Nguyen Van
Thanh, Hoang Thanh Huong, and Phan Van Kiem. Highly oxygenated sterols from the
starfish Archaster typicus. Tp ch Ha hc, 2007, 45(3), 377-381.
499

28. Hp cht 27-norcholestan-3,4,5,6,7,8,14,15,24R-nonol

- Cu trc ha hc:
OH
21

22
20

18
12
11

19
1

10

16
14

15

OH

- Hot tnh sinh hc:

+ Hot tnh khng NF-B:
IC50 = 28,11,98 M
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 4,52
g/ml

OH

HO

23

17

13

OH

26

25

24

OH

OH
OH

OH

- Tinh th hnh kim mu trng: mp 288-290oC

- Cng thc phn t: C26H46O9 (M = 502)
- Ph khi lng: m/z 501,3 [M-H]-, 525,2 [M+Na]+
- c phn lp t loi: Archaster typicus
- c cng b trong cng trnh: Chau Van Minh, Nguyen Xuan Cuong, Nguyen Van
Thanh, Hoang Thanh Huong, and Phan Van Kiem. Highly oxygenated sterols from the
starfish Archaster typicus. Tp ch Ha hc, 2007, 45(3), 377-381.

29. Hp cht ergost-22-en-3,4,5,6,8,14,15,22E,24R,25R,26-nonol

- Cu trc ha hc:

28
21

22
20

18
12
11

19
1

HO

10

25

23

CH 2 OH
OH

27

17

13

OH

26
24

16
14

15

8
7

OH

OH

- Hot tnh sinh hc:

+ Hot tnh khng NF-B:
IC50 = 27,81,87 M
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 3,72
g/ml

OH
OH

OH

- Tinh th hnh kim mu trng: mp 288-290oC

- Cng thc phn t: C28H48O9 (M = 528)
- Ph khi lng: m/z 527,4 [M-H]+, 551,3 [M+Na]+
- c phn lp t loi: Archaster typicus
- c cng b trong cng trnh: Chau Van Minh, Nguyen Xuan Cuong, Nguyen Van
Thanh, Hoang Thanh Huong, and Phan Van Kiem. Highly oxygenated sterols from the
starfish Archaster typicus. Tp ch Ha hc, 2007, 45(3), 377-381.

500

30. Hp cht ergosta-1,3,5,6-tetraol.

- Cu trc ha hc:

28

21
25

20
17

OH
1

27

13

11

19

- c phn lp t loi: Sarcophyton

mililatensis
- Hot tnh sinh hc:
+Hot tnh chng long xng

26
24

18

9
10

HO
OH

Hot lc ca
enzym ALP (%)

0,3
3
30

132,96,623
126,54,246
91,300,766

Hm lng
collagen (%)

S khong ha
(%)

105,61,259
100,03,690
89,634,532

104,501,324
98,123,197
105,82,156

OH

- Tinh th hnh kim mu trng

- Cng thc phn t: C28H50O4, M = 450
- Ph khi lng: m/z: 473,1 [M + Na]+,
450,9 [M +H]+
- c cng b trong cng trnh: Chau Van Minh, Nguyen Xuan Cuong, Tran Anh
Tuan, Eun Mi Choi, Young Ho Kim, and Phan Van Kiem, A new 9,11-secosterol from
the Vietnamese Sea soft coral Sarcophyton mililatensis, increases the function of
osteoblastic MC3T3-E1 cells, Natural Product Comunications (USA), 2007, 2(11),
1095-1100.

31. Hp cht sarcoaldesterol B

- Cu trc ha hc:

28

21

26
24

18

HO
19

20
17

13

11

25

27

- c phn lp t loi: Sarcophyton

mililatensis
- Hot tnh sinh hc:
+Hot tnh chng long xng

10
3

0,3
3
30

HO
OH
OH

Hot lc ca
enzym ALP
(%)
126,31,486
124,80,368
88,542,872

Hm lng
collagen (%)
109,72,302
108,01,012
98,001,801

S khong
ha (%)
100,91,763
104,51,808
97,851,195

- Cht bt mu trng
- Cng thc phn t: C28H50O4, M = 450
- Ph khi lng: m/z: 473,1 [M+Na]+,
433,1 [M-H2O+H]+, 415,1 [M-2H2O+H]+,
397,1 [M-3H2O+H]+
- c cng b trong cng trnh: Chau Van Minh, Nguyen Xuan Cuong, Tran Anh
Tuan, Eun Mi Choi, Young Ho Kim, and Phan Van Kiem, A new 9,11-secosterol from
the Vietnamese Sea soft coral Sarcophyton mililatensis, increases the function of
osteoblastic MC3T3-E1 cells, Natural Product Comunications (USA), 2007, 2(11),
1095-1100.

501

32. Hp cht 2,3,12,21,24R,25-hexahydroxylanosta-8-en

- Cu trc ha hc:
OH

21

OH

18

12
11

HO

HO

16

30

- Tinh th hnh kim mu trng

- Cng thc phn t: C30H52O6 (M
508
- Ph khi lng: m/z 490 [MH2O]+
- c phn lp t loi: Holothuria
scabra

H
28

OH
27

15

26
25

23

20

14

2
10

24

17

22

13

19

OH

29

- c cng b trong cng trnh: Nguyen Van Thanh, Chau Van Minh, Hoang Thanh
Huong, Phan Van Kiem, Steroidal constituents from Holothuria scabra living in
Vietnamese sea, Tuyn tp cc bi bo khoa hc Hi ngh Khoa hc ln th 20 - i hc
Bch Khoa H Ni, 2006, trang 200-204.

33. Hp cht 24R-ergosta-7,22-dien-3,5,6-triol

- Cu trc ha hc:
28
21
18

22
23

20

12
13

19
1

14

10
5

HO

OH

25

16

26

27

17

11

24

15

- Tinh th hnh kim mu trng

- Cng thc phn t: C28H46O3, M =
430.
- Ph khi lng: m/z 412 [MH2O]+
- c phn lp t loi: Holothuria
scabra

7
6

OH

- c cng b trong cng trnh: Nguyen Van Thanh, Chau Van Minh, Hoang Thanh
Huong, Phan Van Kiem, Steroidal constituents from Holothuria scabra living in
Vietnamese sea, Tuyn tp cc bi bo khoa hc Hi ngh Khoa hc ln th 20 - i hc
Bch Khoa H Ni, 2006, trang 200-204.

502

34. Hp cht cholest-7-ene-3,5,6-triol

- Cu trc ha hc:

21

22
20

18

27

13
9

10

5
4

15

- Tinh th hnh kim mu trng

- Cng thc phn t: C27H46O3, M
= 418
- Ph khi lng: m/z 401,2 [MH2O+H]+, 383,2 [M-2H2O+H]+,
365,2 [M-3H2O+H]+
- c phn lp t loi: Gellius
varius

HO

25
17

11
19
1

26

OH
OH

- c cng b trong cng trnh: Nguyn Xun Cng, Nguyn Hi ng, Cng
Thung, Trn Thu Hng, Nguyn Th Kim Thy, Phan Vn Kim v Chu Vn Minh,
Isoprenoids from the sponge Gellius varius living in Vietnamese sea, Tp ch Ha hc,
2008, 46(1), 96-101.

35. Hp cht 6-methoxycholest-7-ene-3,5-diol

- Cu trc ha hc:

21
18

HO

25
27

13
9

10

26

20

17

11
19
1

22

15

- Tinh th hnh kim mu trng

- Cng thc phn t: C28H48O3, M =
432
- Ph khi lng: m/z 415,3 [MH2O+H]+, 397,2 [M-2H2O+H]+, 383,3
[M-2H2O-Me+H]+, 365,3 [M-2H2OMe-H2O+H]+, 431,1 [M-H]+
- c phn lp t loi: Gellius
varius

OH
OCH3

- c cng b trong cng trnh: Nguyn Xun Cng, Nguyn Hi ng, Cng
Thung, Trn Thu Hng, Nguyn Th Kim Thy, Phan Vn Kim v Chu Vn Minh,
Isoprenoids from the sponge Gellius varius living in Vietnamese sea, Tp ch Ha hc,
2008, 46(1), 96-101.

503

VI.8. Lp cht glycolipit

36. Hp cht 1-O-palmitoyl-3-O-[-D-galactopyranosyl-(16)--D-galactopyranosyl]-sn-glycerol
- Cu trc ha hc:
HO

6"

5"

OH
4"

O
1"

OH

6'

2"

3"

5'

OH OH

OH

4'

1'

CH2
2

HC

2'

3'

OH
1'"

CH2 O

OH

16'"

(CH2)14CH3

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng KB: IC50 = 3,51 g/ml
Dng Hep-G2: IC50 = 3,15 g/ml
+ Hot tnh khng VSV kim nh
P. aeruginosa: MIC = 150 g/ml
S. aureus: MIC = 200 g/ml

- Cht bt v nh hnh mu trng

- Cng thc phn t: C31H58O14, M = 654
- Ph khi lng: m/z 653,3 [M-H]+, 677,3
[M+Na]+,
515,2
[M+Na-162]+,
689,4
+
[M+2H2O-H]
- c phn lp t loi: Gellius varius
- c cng b trong cng trnh: Nguyn Xun Cng, Phan Vn Kim v Chu Vn
Minh. Cc hp cht glycolipit t loi hi min cnh xanh Gellius varius sinh sng ti cc
vng bin Vit Nam, Tp ch Khoa hc & Cng ngh, 2007, 45 (2), 41-47.

37. Hp cht (2S)1,2-di-O-palmitoyl-3-O-(6-sulfo--D-quinovopyranosyl)

glycerol
- Cu trc ha hc:
6'

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng FL: IC50 = 4,06 g/ml
Dng Hep-G2: IC50 = 2,89 g/ml
+ Hot tnh khng VSV kim nh
P. aeruginosa: MIC = 100 g/ml
B. subtilis: MIC = 100 g/ml
F. oxysporum: MIC = 50 g/ml
S. cerevisiae: MIC = 200 g/ml

CH2SO3H
5'
4'

OH

O
1'

OH
2'
3'

OH

CH2
2

HC

1'"

CH2 O

16'"

(CH2)14CH3

O 1"
16"
C (CH2)14CH3
O

- Cht bt v nh hnh mu trng

- Cng thc phn t: C41H78O12S, M = 794
- Ph khi lng: m/z 793,6 [M-H]+
- c phn lp t loi: Gellius varius
- c cng b trong cng trnh: Nguyn Xun Cng, Phan Vn Kim v Chu Vn
Minh. Cc hp cht glycolipit t loi hi min cnh xanh Gellius varius sinh sng ti cc
vng bin Vit Nam, Tp ch Khoa hc & Cng ngh, 2007, 45 (2), 41-47.

504

38. Hp cht mui natri ca (2S)1-O-hexadecanoyl-3-O-(6-sulfo--Dquinovopyranosyl) glycerol

- Cu trc ha hc:
6'

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 2,71 g/ml
+ Hot tnh khng VSV kim nh
E. coli: MIC = 200 g/ml
B. subtilis: MIC = 200 g/ml
S. cerevisiae: MIC = 200 g/ml

CH2SO3Na
5'
4'

OH

O
1'

OH
2'

3'

CH2
2

OH

HC

OH
1"

CH2 O

16"

(CH2)14CH3

- Cht bt v nh hnh mu trng

- Cng thc phn t: C25H47NaO11S, M = 578
- Ph khi lng: m/z 555,3 [M-Na]+, 601,2
[M+Na]+
- c phn lp t loi: Gellius varius
- c cng b trong cng trnh: Nguyn Xun Cng, Phan Vn Kim v Chu Vn
Minh. Cc hp cht glycolipit t loi hi min cnh xanh Gellius varius sinh sng ti cc
vng bin Vit Nam, Tp ch Khoa hc & Cng ngh, 2007, 45 (2), 41-47.

VI.9. Cc lp cht khc

39. Hp cht saringosterol
- Cu trc ha hc:

29
28
21

18

25

OH
27

13
9

10

26

20

17

11
19
1

24

14
8

HO

- Cng thc phn t: C29H49O2, M =

428
- Ph khi lng: m/z 411,1 [MH2O+H]+, 393,0 [M-2H2O+H]+
- c phn lp t loi: Xestospongia
testudinaria
- Hot tnh khng vi khun lao:
M. turbeculosis: IC50 = 0,25 g/ml

- Tinh th hnh kim mu trng: mp 160-161oC

- c cng b trong cng trnh: Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu
Phng, V Vn Thnh, Phan Vn Kim v Chu Vn Minh, Nghin cu thnh phn ha
hc loi hi min Xestospongia testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v
Cng ngh, 2007, 45(3), 43-50.

505

40. Hp cht 5,8-epidioxycholest-6-en-3-ol

- Cu trc ha hc:

22

21
18

27

13
9
8

10
3

15

HO

26

25

17

11
19
1

20

- c phn lp t loi: Xestospongia

testudinaria v Gellius varius
- Hot tnh sinh hc:
+ hot tnh khng NF-B
IC50 = 34.7 1.84 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 2,0 g/ml
Dng FL: IC50 = 3,93 g/ml
Dng Hep-G2: IC50 = 2,4 g/ml

- Tinh th hnh kim mu trng: mp 102-105oC

- Cng thc phn t: C27H44O3, M = 416
- Ph khi lng: m/z: 417,0 [M+H]+, 439,0 [M
+Na]+
c cng b trong cc cng trnh:
- Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn
Kim v Chu Vn Minh, Nghin cu thnh phn ha hc loi hi min Xestospongia
testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v Cng ngh, 2007, 45(3), 43-50.
- Nguyn Xun Cng, Nguyn Hi ng, Cng Thung, Trn Thu Hng, Nguyn Th
Kim Thy, Phan Vn Kim v Chu Vn Minh, Isoprenoids from the sponge Gellius
varius living in Vietnamese sea, Tp ch Ha hc, 2008, 46(1), 96-101.

41. Hp cht cholest-7-en-3-on

- Cu trc ha hc:

21

22

18

19

12

11

13
14

2
3

1
10
4

20
17

24
23

26
25
27

16
15

8
7

- Tinh th hnh kim mu trng: mp

147-148oC
- Cng thc phn t: C27H44O, M =
384
- Ph khi lng: m/z 385,2 [M+H]+
- c phn lp t loi: Xestospongia
testudinaria v Archaster typicus

c cng b trong cc cng trnh:

- Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn
Kim v Chu Vn Minh, Nghin cu thnh phn ha hc loi hi min Xestospongia
testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v Cng ngh, 2007, 45(3), 43-50.
- Phan Van Kiem, Nguyen Van Thanh, Nguyen Xuan Cuong, and Chau Van Minh.
Cholest-7-en-3-one, cholesterol and batilol from the starfish Archaster typicus. Tp ch
Ha hc, 2007, 45(4), 513-517.

506

42. Hp cht cholesterol

- Cu trc ha hc:

21

21

20

24

25

18
17

26

19
1
8
3

HO

- Tinh th hnh kim mu trng: mp

148-149oC
- Cng thc phn t: C27H46O,
M=386
- c phn lp t loi: Gellius
varius, Cladiella sp., Xestospongia
testudinaria v Archaster typicus

c cng b trong cc cng trnh:

- Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn
Kim v Chu Vn Minh, Nghin cu thnh phn ha hc loi hi min Xestospongia
testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v Cng ngh, 2007, 45(3), 43-50.
- Phan Van Kiem, Nguyen Van Thanh, Nguyen Xuan Cuong, and Chau Van Minh.
Cholest-7-en-3-one, cholesterol and batilol from the starfish Archaster typicus. Tp ch
Ha hc, 2007, 45(4), 513-517.

43. Hp cht -Adenosine

- Cu trc ha hc:
NH 2

N
N

HO
O
(R)

(R)

OH

OH

- Tinh th hnh kim mu trng: mp 234235oC

- Cng thc phn t: C10H13N5O4, M = 267
- Ph khi lng: m/z 267,8 [M + H]+, 289,8
[M + Na]+
- c phn lp t loi: Holothuria scabra

44. Hp cht thimidine

- Cu trc ha hc:
O
7

CH3

HN3

2
1

5'

HOCH2
O
1'

4'
3'

- Tinh th hnh kim mu trng: mp 186187oC

- Cng thc phn t: C10H14N2O5, M = 242
- Ph khi lng: m/z 264,7 [M+Na]+, 240.8
[M-H]- c phn lp t loi: Xestospongia
testudinaria

2'

OH

- c cng b trong cng trnh: Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu
Phng, V Vn Thnh, Phan Vn Kim v Chu Vn Minh, Nghin cu thnh phn ha
hc loi hi min Xestospongia testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v
Cng ngh, 2007, 45(3), 43-50.

507

45. Hp cht thimine

- Cu trc ha hc:
- Tinh th hnh kim mu trng: mp 326oC
- Cng thc phn t: C5H6N2O2, M = 126
- Ph khi lng: m/z 126,8 [M+H]+, 148,7
[M+Na]+, 124,7 [M-H]- c phn lp t loi: Xestospongia
testudinaria

O
7

CH3

HN 3

2
1

N
H

- c cng b trong cng trnh: Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu
Phng, V Vn Thnh, Phan Vn Kim v Chu Vn Minh, Nghin cu thnh phn ha
hc loi hi min Xestospongia testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v
Cng ngh, 2007, 45(3), 43-50.

46. Hp cht batyl alcohol (batilol)

- Cu trc ha hc:
1

- Tinh th mu trng: mp 70-71oC

- Cng thc phn t: C21H44O3, M = 344
- Ph khi lng: m/z 345,1 [M+H]+
- c phn lp t loi: Xestospongia
testudinaria, Cladiella sp v Archaster
typicus

OH
HO

1'

18'

CH2(CH2)16CH3

c cng b trong cc cng trnh:

- Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn
Kim v Chu Vn Minh, Nghin cu thnh phn ha hc loi hi min Xestospongia
testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v Cng ngh, 2007, 45(3), 43-50.
- Phan Van Kiem, Nguyen Van Thanh, Nguyen Xuan Cuong, and Chau Van Minh.
Cholest-7-en-3-one, cholesterol and batilol from the starfish Archaster typicus. Tp ch
Ha hc, 2007, 45(4), 513-517.

47. Hp cht chimyl alcohol

- Cu trc ha hc:
1

- Tinh th mu trng: mp 64-65oC

- Cng thc phn t: C19H40O3, M=316
- Ph khi lng: m/z: 317,1 [M+H]+
- c phn lp t loi: Xestospongia
testudinaria v Cladiella sp

OH
HO

1'

16'

CH2(CH2)14CH3

c cng b trong cc cng trnh:

- Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn
Kim v Chu Vn Minh, Nghin cu thnh phn ha hc loi hi min Xestospongia
testudinaria thu thp ti Vit Nam. Tp ch Khoa hc v Cng ngh, 2007, 45(3), 43-50.
- Chau Van Minh, Phan Van Kiem, and Nguyen Xuan Cuong, New 9,11-secosterol
glycoside and ceramide from a soft coral Cladiella sp. living in Vietnamese sea, Asean
Journal on Sicience and Technology for Development, 2007, 24(3), 275-283.

508

48. Hp cht trans-phytol

- Cu trc ha hc:
17

18

19

11

15

20

OH

16

- Cht du khng mu
- Cng thc phn t: C20H40O, M = 296
- c phn lp t loi: Gellius varius
- c cng b trong cng trnh: Nguyn Xun Cng, Nguyn Hi ng, Cng
Thung, Trn Thu Hng, Nguyn Th Kim Thy, Phan Vn Kim v Chu Vn Minh,
Isoprenoids from the sponge Gellius varius living in Vietnamese sea, Tp ch Ha hc,
22008, 46(1), 96-101.

49. Hp cht metyl este ca axit (5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic

- Cu trc ha hc:
O
11

20

17

14

- Cht du khng mu
- Cng thc phn t: C21H32O2, M = 316
- Ph khi lng: m/z 317,1 [M+H]+, 315,0 [M-H]+
- c phn lp t loi: Gellius varius
- c cng b trong cng trnh: Nguyn Xun Cng, Phan Vn Kim v Chu Vn
Minh. Cc hp cht glycolipit t loi hi min cnh xanh Gellius varius sinh sng ti cc
vng bin Vit Nam, Tp ch Khoa hc & Cng ngh, 2007, 45 (2), 41-47.

50. Hp cht glycerol 1-(5Z,8Z,11Z,14Z-eicosatetraenoate)

- Cu trc ha hc:

HO

OH
2

O
5'

8'

1'

O
20'
11'

14'

- Cht du khng mu
- Cng thc phn t: C23H38O4 (M=378)
- Ph khi lng: m/z 379,2 [M+H]+, 361,2 [M-H2O+H]+, 347,2 [M-H2O-CH2+H]+, 329,1
[M- H2O-CH2-H2O+H]+, 287,2 [M-Glycerol+H]+, 377,1 [M-H]+
- c phn lp t loi: Gellius varius
509

- c cng b trong cng trnh: Nguyn Xun Cng, Phan Vn Kim v Chu Vn
Minh. Cc hp cht glycolipit t loi hi min cnh xanh Gellius varius sinh sng ti cc
vng bin Vit Nam, Tp ch Khoa hc & Cng ngh, 2007, 45 (2), 41-47.

51. Hp cht 1-heptadecanol

- Cu trc ha hc:
1

16
17

HO

14

CH3

- Tinh th mu trng: mp 59,5-60,0oC

- Cng thc phn t: C17H36O, M=256
- Ph khi lng: m/z 279.0 [M+Na]+, 255.0 [M-H]-, 237.0 [M-H2O-H]- c phn lp t loi: Cladiella sp

VI.10. nhn xt
Trong khun kh ti KC09.09/06-10 chng ti thu thp c 101 mu
sinh vt bin ca ba nhm l: san h mm, da gai v hi min. Trn c s kt qu
sng lc hot tnh sinh hc, tr lng, kh nng thu mu v tra cu cc ti liu
nghin cu cng b trn th gii, 07 i tng sinh vt bin c la chn thu
mu lng ln v tin hnh nghin cu ha hc theo nh hng hot tnh sinh hc
l: San h mm Sarcophyton mililatensis v Cladiella sp., hi sm Holothuria
scabra, sao bin Archaster typicus, hi min Xestospongia testudinaria, Gellius
varius v Ianthella sp.
Nghin cu thnh phn ha hc theo nh hng hot tnh sinh hc ca 07
i tng sinh vt bin phn lp, xc nh cu trc, v th hot tnh sinh hc
c 51 hp cht (18 cht mi) trong c: 07 hp cht ditecpen dng cembranoit
(05 cht mi), 03 hp cht 9,11-secosteroit (02 cht mi), 06 hp cht holothurin
(03 cht mi), 03 hp cht cerebrosit v ceramit (02 cht mi), 02 axit bo a ni
i v ni ba b brm ha mi, 05 hp cht steroit mang cu trc vng propan
mch nhnh (03 cht mi), 09 hp cht steroit mang nhiu nhm hydroxy trong
phn t, 03 hp cht glycolipit v 13 hp cht khc.
T l cc cht mi trong tng s cht phn lp c thuc cc nhm cht
ditecpen dng cembranoit, 9,11-secosteroit, holothurin, axit bo a ni i - ni ba
b brm ha, steroit c cu trc vng propan mch nhnh, cerebrosit v ceramit l
rt cao. y u l cc lp cht c cu trc c th v gi nhiu chc nng sinh hc
quan trng trong c th sinh vt cng nh th hin nhiu hot tnh qu bu. Cc
cembranoit gi vai tr phng v ha hc ca cc sinh vt chng li k th. Chng
thng th hin nhiu hot tnh sinh hc qu bu nh bo v thn kinh, khng vi
sinh vt, khng vim v khng ung th. Cc 9,11-secosterol tn ti vi s lng
khng nhiu trong t nhin, chng thng th hin hot tnh gy c t bo v
chng tng sinh rt nhiu dng t bo ung th khc nhau nh C6 (t bo lioma
chut), J774 (bch cu n nhn to v i thc bo chut), LnCap (t bo ung th
tuyn tin lit), Calu-3 (ung th phi).... Cc holothurin thng c ph hot
510

tnh rng nh: gy c t bo, khng nm, tan mu, iu bin min dch... Cc
holothurin th hin nhiu hot tnh qu bu nn c gi tr rt cao. n c nh hp
ct holothurin A c hng Sigma rao bn vi gi 5000 USD/1g. V phng din
sinh hc, mt s cerebrosit c chng minh gi vai tr h tr cu trc v xc
nh kt cu ca cc mng t bo, chng hot ng nh cc cht iu tit ca nhiu
qu trnh sinh hc nh qu trnh hot ha, qu trnh dnh kt t bo, trao i thng
tin ni bo v pht trin ca t bo. Hn na, cc cerebrosit trong mng t bo gi
vai tr sinh hc quan trng l cc khng nguyn b mt v cc th th. c bit,
ngy cng c nhiu bng chng ch ra rng cc cerebrosit c cc chc nng sinh hc
rt a dng, hu ht cc chc nng ny u lin quan n c tnh lng cc ca
phn t. Chng hn nh, nhn t pht trin thn kinh (NGF - Nerve growth factor),
mt i phn t sinh hc l nhn t u tin c xc nh r nht, chng iu tit
s pht trin, bit ha v sng st ca cc nron v c coi l mt ng vin nng
k pht trin cc thuc tr bnh Alzheimer (bnh qun). Tuy nhin, phn t NGF
qu ln v mt kch thc v qu a nc i qua cc vch ngn mu no, m
li l con ng an ton duy nht a cc NGF vo trong no tht. Do , cc
cht phn t lng thp c tc dng tng t cc NGF (chng hn nh cc
cerebrosit) l cc ng vin y ha hn cho vic tr bnh Alzheimer. Cc hp cht
axit mch di a ni i v ni ba b brm ha thng c hot tnh khng sinh v
gy c t bo. c bit, mt s hp cht th hin hot tnh c ch enzym HIV
protease, mt enzym thit yu cho s ti bn ca HIV. Thm vo , chng cn c
hot tnh km hm tc nhn i khng lin kt vi th th Adenosine. Khi th th
ny c hot ha s mang n cc hiu qu tch cc nh gim nhp tim, bo v no
v tiu m.
Cc nghin cu trong khun kh ti cho thy, cc hp cht cembranoit v
9,11-secosteroit cn th hin hot tnh chng long xng thng qua cc c ch kch
thch s pht trin v bit ha ca nguyn bo xng. Kt qu ny cho thy cc hp
cht hot ng nh mt tc nhn iu bin chn lc (SERM) v iu bin qu
trnh ri lon xng ph n thi k mn kinh. Cc hp cht ny c phn lp vi
hm lng kh cao, iu ny gi m cho nhng nghin cu su hn v cc hp cht
ny theo hng ng dng trong vic phng v iu tr bnh long xng, mt cn
bnh rt ph bin hin nay (c bit l trong cng ng ngi cao tui). Ngoi ra,
cc hp cht 9,11-secosterol, steroit mang nhiu nhm hydroxy, cerebrosit v
ceramit cn th hin hot tnh khng yu t phin m NF-B. Yu t ny iu tit s
biu hin ca nhiu gen tham gia vo cc p ng min dch v vim. N lm tng
s biu hin ca cc gen quy nh nhiu cytokine, cc enzym v cc phn t dnh
kt trong cc bnh vim mn tnh. NF-B c hot ha bi nhiu yu t khc nhau
(virus, cc cht xy ha v cc khng nguyn) lm tng p ng vim. S hot ha
ny li dn ti s biu hin phi hp ca nhiu gen m ha cc protein (nh cc
cytokine, chemokine, enzym v cc phn t dnh kt) tham gia vo s tng hp cht
trung gian v ko di thm p ng vim. V vy, NF-B l mt ch hin nhin cho
cc phng php iu tr khng vim mi. Glucococticoit l cc cht c ch NF-B
hu hiu nhng chng c nhiu tc dng ph ti ni tit v chuyn ha khi c s
dng vi liu php iu tr ton thn. Cc tc dng ph ny s khng xy ra khi s

511

dng cc cht c ch NF-B c hiu hn. V vy, vic tm kim v nghin cu k

lng cc hp cht thin nhin c tc dng c ch c hiu NF-B v ang thu
ht mnh m s quan tm v u t nghin cu ca cc nh khoa hc trn ton th
gii.
Nh vy, cc kt qu nghin cu thnh phn ha hc v hot tnh sinh hc
t c trong ti mang ngha khoa hc rt cao (c bit l trong lnh vc ha
hc cc hp cht thin nhin bin, mt lnh vc nghin cu cn rt non tr Vit
Nam) v m ra nhiu hng nghin cu ng dng pht trin cc dc t mi t
ngun dc liu bin di do ca t nc trong tng lai. Cc kt qu c
ng trn 17 bi bo khoa hc trong c 11 bi trn cc tp ch chuyn ngnh
quc gia v 06 bi bo quc t (05 bi ng v 01 bi gi ng. Ngoi ra ti
ng k c 02 bng c quyn sng ch ti cc s hu tr tu Vit Nam (01
bng c cp). iu ny s gp phn a cc nghin cu v ha hp cht thin
nhin bin ca nc ta ln ngang tm cc nc trong khu vc cng nh quc t
ng thi bo h c cc nghin cu ca t nc.

512

C. Cc nghin cu thm d v rong, to v vi sinh

vt bin
Chng VII. Nghin cu thm d kh nng khng vim
t rong, to bin vit nam.
VII.1. M u
Vim hin ang c coi l cn bnh ph bin, cc loi thuc chng vim
trn th trng ngy nay l cc thuc NSAID (cc thuc khng vim khng thuc
dng steroit). Cc thuc ny hu ht hot ng da trn c ch km hm hot
ng ca cc enzyme xc tc cho qu trnh to cht gy vim nh COX-1 v
COX-2 (cyclooxygenase), lipogenase (cc enzyme xc tc cho vic to nn cc
prostaglandin v leukotrien nhn t quan trng trong qu trnh gy vim, t axit
arachidonic), PLA2 (phospholipase A2) - enzyme xc tc qu trnh to axit
arachidonic. Cht km hm PLA2 c ngun gc t nhin u tin c bit l mt
sesterterpen c tn l manoalide (IC50 = 3,9 M), c tch t loi bt bin
Luffatiella variabilis cho thy hot ng khng vim v gim au rt cao, hin
ang c th nghim lm sng trn pha 1. Mc d cha c mt trn th trng
thuc, nhng manoalide c coi nh mt cht chun trong hot ng km hm
PLA2 [283-286]. Mt nhn t khng vim ng quan tm khc l cc
Pseudopterosin, c phn lp t loi san h sng Pseudopterogorgia
elisabethae. Chng hot ng khng vim rt cao trn c 2 c ch km hm PLA2
cng nh 5-Lypoxygenase. Mt trong s cc dn xut ca pseudopterosin hin
ang c th nghim lm sng v chun b a ra th thng trong thi gian ti.
Ngoi ra, pseudopterosin ang c bn rng ri trn th trng di dng m
phm v kem chng nng rt thnh cng [287-288]. Nh vy, chng vim,
ngoi vic cn tng cng h min dch ca c th, cn cn thit phi phng nga
vim.
Gn y, hot ng khng vim ca cc axit bo khng bo ho a ni i
omega 3 (-3 PUFAs) in vivo v in vitro c khng nh [289-291]. -3
PUFAs c th iu chnh chng vim nhim v s hnh thnh cc cytokine tin
vim nhim. Chng cng iu chnh s chuyn ha prostaglandin v lm gim
triglycerids, h thp cholesterol v c cc c tnh khng huyt khi v khng
vim. Cc eicosanoid v cc dn xut ca chng c ch nhiu trong vic pht
trin cc th h thuc khng vim mi. Mt s to bin c th dng lm thc
phm cha -3 PUFAs vi t l cao. Nhiu nghin cu gn y c tnh rng
vic b sung -3 PUFAs c tc dng trong vic chng li nhiu bnh vim
nhim. Tc dng khng vim ca cc -3 PUFAs ch yu lin quan n s cnh
tranh ca chng nh l cc c cht cho cc enzyme COX v LOX dn n s to
thnh cc prostaglandin v leukotrien t hot tnh. Cc cht khng vim c bn
cht sterol glycoside c tch chit t Undaria pinnatifida v Enteromorpha
linza [292], cc axit bo EPA, DHA, n-6DPA c tc ng ti h thn kinh v tim
mch c tch chit t vi to bin Labyrinthula spp. v Schizochytrium spp.,
cng c vai tr trong phng nga vim [293-295].

513

Lyngbyatoxin A v debromoaplysiatoxin l hai cht c hot tnh khng

vim cao, nhng cu trc khc nhau c phn lp t cc chng to lam gy c
l Lyngbya mausculata Hawaii [296] v anatoxin-A t Anabaena ciecinalis
[297] To lc Ulva lactuca c cha cht khng vim [298]. Cc cht chng oxy
ho c s dng trong vic ngn chn hnh thnh cc gc t do v cc dng hot
tnh ca oxygen. Mt s hp cht c hot tnh km hm hot ng to gc t do
superoxit - mt yu t lin quan n cn bnh ung th, vim nhim v lo ho.
Do , vic pht trin cc nhn t km hm qu trnh to superoxit c th gip
ngn chn cc bnh trn.
vng bin Vit Nam, rong bin rt a dng, phong ph v thnh phn
chi, loi v t lu c bit l ngun nguyn liu qu gi v cc cht c hot
tnh sinh hc. Nhng hp cht t nhin ny c nhng tnh cht c hiu. Tuy
nhin, gi tr s dng trong c gi tr v dc hc cn cha hiu bit c y
. Trong s cc hot cht t rong bin c th khai thc v a vo s dng lm
thuc phi k n cc cht c hot tnh khng vim v cc axt bo khng bo
ho a ni i LCFUFA, (3) (DHA) v 3 (n-6DPA).
VII.2. KT QU V THO LUN
VII.2.1.Thu thp v nh tn s b 19 loi Rong to bin Vit Nam (tp
trung cc loi thuc Rong , Nu v Lc) cho phn tch hot tnh khng
vim
19 Loi rong c thu mt s tnh pha Bc (Hi Phng), min trung
(Tha Thin, Hu) v min Nam (Nha Trang, Khnh Ho, Ninh Thun), Vit
Nam (bng II.2.4.1). Cc mu rong sau khi thu ngoi t nhin c nh tn
khoa hc, sau ra sch bng nc bin v nc ngt, phi kh nhit
phng n m 20- 30 % v nghin thnh bt. Bt rong c bo qun -200C.

Hnh VII.2.1. Bt rong ca cc loi rong nghin cu: A/ Undaria pinnatifida, B/

Enteromorpha linza, C/ Ulva reticulata, D/ Sargassum swartzii, E/ Gracilaria asiatica,
F/ Gracilaria blodgettii, G/ Sargassum polycystum.
514

VII.2.2. Hot tnh ca cht khng vim c chit t Undaria pinnatifida

bng cc loi dung mi khc nhau
Kt qu phn tch hot tnh khng vim t Undaria pinnatifida c chit
bng cc loi dung mi khc nhau c ch ra trn bng VII.2.2.

Hnh VII.2.2.a. Quy trnh tch dch chit c hot tnh

khng vim t bt cc loi rong bin nghin cu
Bng VII.2.2. Hot tnh ca dch chit khng vim c chit t
pinnatifida bng cc loi dung mi khc nhau
Dung mi

Gi tr ph n

Gi tr ban

0,03 0,00
0,17 0,02
0,03 0,00
0,03 0,00
0,13 0,00
0,12 0,00
0,27 0,00
0,12 0,02
0,62 0,02
0,12 0,00
0,03 0,00

Phn trm
c ch
96
79
96
96
84
85
67
85
23
85
96

Acetonitrile
Acetone
Benzene
Chloroform
Cyclohexane
Ethyl axetat
Ethyl alcohol
te
Hexan
Metanol
Methylene
chloride
Nc kh trng
Nc si
i chng

0,70 0,12
0,80 0,00
0,81 0,04

14
1
0

0,010,00
0,03 0,00
0,04 0,00

Undaria

Phn trm c
ch
96
89
85

0,06 0,01

78

0,14 0,04

48

0,10 0,00
0,06 0,00

63
78

0,17 0,00
0,16 0,02
0,27 0,01

37
4
0

Ghi ch: *: khng xc nh; i chng: khng c bi dch chit c hot tnh khng vim. Mi gi tr i din trung bnh S.D. ca t 6 n 7 con chut. Sai khc c ngha thng k
P<0,05

515

5
6

Hnh VII.2.2.b. Dch chit i din ca mt s loi rong nghin cu: 1. Undaria
pinnatifida, 2. Enteromopha linza, 3. Gracilaria asiatica, 4. Gracilaria blodgettii,
5. Sargassum polycystum, 6. Sargassum swartzii, 7. Ulva reticulata
Kt qu trong bng VII.2.2 cho thy, hot tnh khng vim trong cc dch
chit t U. pinnatifida vi cc dung mi khc nhau l khc nhau. Hot tnh khng
vim cao nht khi rong c chit vi acetonitrile, benzen v metanol. Tng ng
vi phn trm c ch ph n v ban trong dch chit vi acetonitrile l 96% v
96%; trong benzen l 96% v 85%, v trong metanol l 85% v 78%, tng ng
tng gi tr. Tuy nhin, tch mt lng ln cht khng vim vi gi thnh r v
cht lng tt phc v cho cc th nghim tip theo chng ti chn dung mi
tch chit l metanol. Qui trnh tch chit cht khng vim bng metanol c
trnh by trong hnh VII.2.2.a v b.
VII.2.3. Sng lc hot tnh khng vim trong cc loi rong bin Vit Nam
Sau khi chn c dung mi cn thit cho qu trnh tch chit cht c
hot tnh khng vim, chng ti tin hnh sng lc hot tnh ny trong 19 loi
rong bin Vit Nam v s dng 2 loi (Enteromorpha linza v Undaria
pinnatifida) ca Hn Quc lm mu th so snh vi mu rong ca Vit Nam. Kt
qu sng lc hot tnh khng vim trong 19 loi rong bin nghin cu c ch ra
trong bng VII.2.3.
Bng VII.2.3. Sng lc hot tnh khng vim trong cc loi rong bin Vit Nam
c dng trong nghin cu
TT
1
2
3
4
5
6
7
8

Tn loi
RONG LC
Codium arabicum
Enteromorpha compressa
Enteromopha linza
Ulva reticulata (1)
Ulva reticulata (2)
RONG NU
Dictyota dichotoma
Sargassum polycystum
S. microcustum

516

c ch ph n

c ch ban

++
+++
+++
++
++++

++
++
+++
++
++++

+++
+++
+++

++
+++
++

9
10
11
12
13
14
15
16
17
18
19
20
21
22
23

S. mcclurei
S. denticarpum
S. swartzii
S. duplicatum
S. binderi
Undaria pinnatifida
RONG
Gracilaria blogettii
Gracilaria asiatica
G. arcuata
G. eucheumoides
G. salicornia
Hypnea valentina
Porphyra crispata
P. suborbiculata
i chng*

+++
+++
+++
+++
+++
++++

+++
+++
++
+++
+++
++++

++
+
+
+
++
+++
++
+++
0

++
+
+
++
++
++
++
++
0

Ghi ch: * i chng : khng c bi dch chit c hot tnh khng vim; + (c ch <25%); ++ (c ch
t 25 n 50%); +++ (c ch t 50 n 75%) v ++++ (c ch >75%).

Kt qu trnh by trn bng VII.2.3 cho thy tt c cc loi rong nghin

cu u c hot tnh khng vim. Cc loi rong bin khc nhau th hot tnh
khng vim cng khc nhau. Trong , Ulva reticulata v Undaria pinnatifida
cho hot tnh khng vim cao nht (chim trn 75% gi tr c ch ph n v ban
), sau n n cc loi Enteromopha linza v Sargassum (S. duplicatum, S.
polycystum, S. mcclurei, S. denticarpum, S. swartzii, S. binderi) (50-70% gi tr
c ch ph n v ban ). Hot tnh khng vim thp nht l trong cc loi
Gracilaria arcuata, G. eucheumoides v G. asiatica (nh hn 20% gi tr c ch
ph n v ban ).
VII.2.4. nh hng ca thi gian iu tr vim bng dch chit t Undaria
pinnatifida ln kh nng khng vim tai chut
Dch chit ca Undaria pinnatifida (Hn Quc) c pha trong Ethanol
nng 0,08 mg/l. Kh nng khng vim ca dch chit ny c xc nh sau
cc thi gian iu tr 30 pht, 1 v 2 gi. Kt qu v nh hng ca thi gian iu
tr vim t dch chit ca Undaria pinnatifida ln kh nng khng vim c ch
ra trn bng VII.2.4.
Bng VII.2.4. nh hng ca thi gian iu tr vim t dch chit ca Undaria
pinnatifida ln kh nng khng vim tai chut*
Thi gian iu tr (gi)
% c ch ban
% c ch ph n
0,5
20,949.97
72,9617,77
1
78,0116,96
85,638,57
2
56,224,21
14,075,86
Ch thch: * - Mi gi tr i din trung bnh S.D. ca t 6 n 7 con chut. Sai khc c
ngha thng k P<0,05.

517

Kt qu ch ra trong bng VII.2.4 cho thy sau 0,5 n 2 gi iu tr bng dch

chit Undaria pinnatifida u c hot tnh c ch ban . Tuy nhin, ch c phn
trm c ch ban trong l chut c iu tr sau 1h l c s sai khc c
ngha thng k khi so snh vi i chng. Phn trm c ch ph n ca dch chit
Undaria pinnatifida cc thi gian iu tr khc nhau cng khc nhau. Kh nng
c ch ph n ca dch chit Undaria pinnatifida t cao nht ti thi im sau 1
gi iu tr (85,63%). Sau thi gian iu tr 2 gi, kh nng c ch ph n v ban
trong dch chit ny gim xung ng k. Do , chng ti chn thi gian
iu tr vim bng dch chit c hot cht khng vim t rong bin cho cc th
nghim tip theo l sau 1 ting.
VII.2.5. nh hng ca cc nng dch chit trong cc loi rong bin
nghin cu ln kh nng khng vim tai chut
Da trn kt qu sng lc cc loi rong c hot tnh khng vim (bng
VII.2.3), chng ti chn ra mt s loi nh l Undaria pinnatifida, Gracilaria
blodgettii, Ulva reticulata, Sargassum swartzii v S. polycystum nghin cu
su hn v nh hng ca chng cc nng dch chit khc nhau ln kh
nng khng vim trn tai chut. Kt qu c ch ra trn bng VII.2.5.
Bng VII.2.5. nh hng ca cc nng dch chit ca cc loi rong bin
nghin cu ln phn trm c ch ban v ph n tai chut
Nng mg/l
Rong bin

U. pinnatifida
Ulva reticulata
S. swartzii
S. polycystum
G. blodgettii
i chng*

Phn trm c ch ban

Phn trm c ch ph n ph n

0,04

0,08

0,16

0,04

0,08

0,16

29,01
5,47
14,71
4,79
18,48
1,91
27,87
0,34
9,80
8,96
0,00

78,56
14,56
80,40
1,55
58,83
0,46
55,25
3,61
26,23
8,94
0,00

67,34
9.45
104,66
3,38
48,47
5,55
38,17
8,89
3,68
8,96
0,00

55,06
5,86
37,95
0,09
32,59
0,80
0,00
0,32
4,99
4,19
0,00

85,46
4,80
75,35
4,83
60,17
5,86
55,06
5,94
6,09
7,31
0,00

82,72
4,01
63,71
3,82
44,38
6,23
49,16
5,73
28,25
9,44
0,00

* i chng: khng c bi dch chit c hot tnh khng vim; Mi gi tr i din trung bnh
S.D. ca t 6 n 7 con chut. Sai khc c ngha thng k P<0,05.

Kt qu trong bng VII.2.5 cho thy phn trm c ch ca dch chit Undaria
pinnatifida, Gracilaria blodgettii, Ulva reticulata, Sargassum swartzii v S.
polycystum cc nng khc nhau cng khc nhau.Trong phn ln cc loi
nghin cu gi tr ny tng dn theo nng .
Khi so snh dch chit thu c t cc loi rong nghin cu nng
0,08ug/ml, chng ti thy rng dch chit ca Ulva reticulata v Undairia
pinnatifida c hot tnh khng vim cao nht (th hin phn trm gi tr c ch
ban t n > 80% v c ch ph n > 75%). Tip theo l hai loi Sargassum
518

swartzii v S. polycystum c thu ven b bin Bi Tin, Vnh Ha, Nha Trang,
Khnh Ha v do cng ty TNHH Vit i cung cp u c hot tnh khng vim
gn tng ng nhau (phn trm gi tr c ch ban r, 83 v 55,25%; c ch
ph n - 60,14 v 55,06%, tng ng tng loi mt). Cn loi rong cu Tht - G.
blogettii c hot tnh khng vim thp nht (26,23 v 6,09%).
Trn c s cc s liu thu c trn, chng ti chn ra hai loi c hot tnh
khng vim cao v tr lng ln l Ulva reticulata (thu thp ti Nha Trang,
Khnh Ha) v S. swartzii tch chit lng ln dch chit khng vim. Dch
chit ca hai loi ny sau , c s dng cho cc nghin cu su hn v th
hot tnh khng sinh trn cc chng vi sinh vt kim nh v khng vim trn
chut (th c tnh cp, tc dng gim au bng axit axetic, gim au mm nng,
gim vim mn bng u ht, gy vim cp mng bng bng carageenane).

Hnh VII.2.5. nh quy trnh tch lng ln dch chit c hot tnh khng vim
t hai loi Ulva reticulata v Sargassum swartzii
VII.2.6. Kt qu th hot tnh vi sinh vt v nm trn cc chng vi sinh vt
v nm kim nh
Kt qu th hot tnh khng sinh trn cc chng vi sinh vt kim nh
Escherichia coli (Ec); Pseudomonas aeruginose (Pa); Bacillus subtilis (Bs) v
Staphylococcus aureus (Sa) v nm kim nh Candida albicans (Ca) c ch ra
trn bng VII.2.6.
Bng VII.2.6. Kt qu th hot tnh khng sinh trn cc chng vi sinh vt v nm
kim nh
STT
1
2

Tn mu
Ulva reticulata
Sargassum
swartzii

Tn cc chng vi sinh vt v nm kim nh

Pa
Bs
Sa
Ca
Ec
IC50g/ml IC50g/ml IC50g/ml IC50g/ml IC50g/ml
62,15
>256
>256
>256
>256
>256

>256

519

>256

56,53

>256

Kt qu trn bng VII.2.6 cho thy, Ulva reticulata v Sargassum swartzii c

hot tnh khng chng Bacillus subtilis (vi gi tr IC50 = 62,15 g/ml) v
Staphylococcus aureus (IC50= 56,53 g/ml), tng ng tng loi mt
VII.2.7. Th nghim c tnh cp
Sau khi cho chut ung dch chit Sargassum swartzii v Ulva reticulata
n liu cao nht (66g/kg), chng ti thy rng chut sau ung thuc th vn hot
ng, n ung v i li bnh thng, phn v nc tiu ca chut khng c g bt
thng. Sau 72 gi, khng c chut no cht. V khng th tng th tch theo
ng ung hn c na (th tch thuc ti a chut c th dung np) nn cha
tnh c LD50 ca cao khng vim Sargassum swartzii v Ulva reticulata trn
chut nht trng theo ng ung.
VII.2.8. Tc dng gim au
* Theo phng php qun au axit axetic (Koster): nh hng ca dch
chit Sargassum swartzii (S) v Ulva reticulata (U) ln s cn au ca chut
c ch ra trn bng VII.2.8.a.
Bng VII.2.8.a. nh hng ca dch chit Sargassum swartzii (S) v Ulva
reticulata (U) ln s cn qun au ca chut.
S cn qun au ca chut (con /5 pht)
L chut nghin cu
0-5 pht

5-10 pht

10-15
pht

15-20
pht

20-25
pht

i chng m (ung
nc ct)
i chng dng
(ung aspirin
100mg/kg)
Dch chit S liu thp
250mg/kg

5,60,52

18,24,71

16,53,75

13,03,30

9,12,51

1,20,42
P1-2<0,001

11,63,63
P1-2<0,001

15,53,47
P > 0,05

12,12,77
P > 0,05

6,31,95
P > 0,05

5,21,55
P > 0,05

11.74,06
P > 0,05

10,43,13
P1-2<0,001

5,71,95
P1-2<0,001

4,61,35
P1-2<0,001

Dch chit S liu cao

500mg/kg

2,90,57
P1-2<0,001

8,81,93
P1-2<0,001

8,93,11
P1-2<0,001

5,81,23
P1-2<0,001

3,10,88
P1-2<0,001

Dch chit U liu thp

250mg/kg

6,01,15
P > 0,05

12,13,87
P < 0,05

13,82,57
P > 0.05

11,43,37
P > 0,05

7,02.21
P > 0,05

Dch chit U liu cao

250mg/kg

6,21,62
P > 0,05

8,81,93
P < 0,05

13,32,45
P < 0,05

7,31,77
P < 0,05

6,21,62
P < 0,05

Kt qu trn bng VII.2.8.a. cho thy, dch chit Sargassum swartzii c 2

liu thp v cao u lm gim r rt s cn au ca chut so vi l i chng m
cc thi im 5-10, 10-15, 15-20sau khi gy au cho chut bng axit axetic
(P<0,05 - 0,001). Liu 500mg/kg c tc dng tt hn v mnh hn so vi aspirin
liu 100mg/kg.

520

Dch chit Ulva reticulata liu 500mg/kg cng lm gim s cn au ca

chut v tng ng vi Aspirin liu 100mg/kg.
* Theo phng php gy au bng mm nng: nh hng ca dch chit
Sargassum swartzii (S) v Ulva reticulata (U) ln thi gian phn ng vi nhit
ca chut nht c ch ra trn bng VII.2.8.b.
Bng VII.2.8.b. nh hng ca dch chit Sargassum swartzii (S) v Ulva
reticulata (U) ln thi gian phn ng vi nhit ca chut nht
Thi gian phn ng vi nht (giy)

L chut nghin cu

T0

T1

P trc-sau

i chng m (ung nc ct)

i chng dng (morphin liu 10
mg/kg)
Dch chit S liu 250mg/kg

13,67 4,01

12,42 3,71

> 0,05

14,08 2,01

21,99 6,93

< 0,05

14,37 3,90

16,22 3,40

> 0,05

Dch chit S liu 500mg/kg

12,61 2,60

16,5 4,80

< 0,05

Dch chit U liu 250mg/kg

15,39 3,81

14,96 4,50

> 0,05

Dch chit U liu 500mg/kg

13,27 2,06

17,87 5,88

< 0,05

Kt qu bng VII.2.8.b cho thy, dch chit Sargassum swartzii v Ulva

reticulata liu 500mg/kg lm gim au r rt thng qua vic lm ko di thi
gian tc dng phn ng vi nhit khi so snh gia trc v sau khi dng thuc
th (P < 0,05). Tc dng ca hai dch chit ny yu hn morphin liu 10mg/kg.
VII.2.9. Tc dng chng vim cp
* Trn m hnh gy ph chn chut bng carrageenin: Tc dng chng vim
ca dch chit t Sargassum swartzii (S) v Ulva reticulata (U) trn m hnh gy
ph chn chut c ch ra trn bng VII.2.9.a.
Bng VII.2.9.a. Tc dng chng vim ca dch chit t Sargassum swartzii (S) v
Ulva reticulata (U) trn m hnh gy ph chn chut
Sau 2 gi (V1)
L

ph
(%)

i chng
Indometacin
(25 mg/kg)

% gim
ph so
vi i
chng

40,44

ph(%)

% gim
ph so vi
i chng

53,90
12,25

14,06
24,92
14,43

Sau 4 gi (V2)

38,38
p1-2< 0,05

24,92
14,43

Sau 6 gi (V3)
ph
(%)

% gim
ph so vi
i chng

55,97
14,12
53,77
P1-2< 0,001

521

37,31
16,42

Sau 24 gi (V4)
ph
(%)

% gim
ph so vi
i chng

28,97
11,96
33,34
P1-2< 0,05

19,37
11,24

33,14
P1-2< 0,05

Dch chit S
(175mg/kg)

26,87
12,08

33,56

Dch chit S
(350mg/kg)

25,03
10,29

38,11

Dch chit U
(175mg/kg)

40,52
20,49

Dch chit U
(350mg/kg)

37,31
21,56

7,74

p1-2< 0,05

P1-2< 0,05

P1-2> 0,05

P1-2> 0,05

45,43
13,24

15,71

44,20
16,58

17,99

64,48
16,86

66,51
33,28

P1-2 > 0,05

P1-2> 0,05

P1-2> 0,05

P1-2> 0,05

40,50
11,32

27,64

40,08
9,0

28,39

51,52
16,04

7,95

55,03
25,25

1,68

P1-2< 0,05

P1-2< 0,05

P1-2> 0,05

P1-2> 0,05

13,87
6,86

52,12

15,69
4,71

45,84

17,56
8,67

4,87

22,71
10,07

21,61

P1-2< 0,05

P1-2< 0,05

P1-2> 0,05

P1-2> 0,05

Kt qu bng VII.2.9.a cho thy, dch chit Sargassum swartzii c 2

liu 175mg/kg v 350mg/kg c tc dng chng vim r rt thng qua lm gim
th tch chn chut gy ph bng carrageenin cc thi im 2, 6 v 24 gi (P<
0,05). Tc dng chng vim ca dch chit t Sargassum swartzii (52,12% liu
175mg/kg v 45,84% liu 350mg/kg) mnh hn khi so snh vi Indometacin
liu 25mg/kg (33,145%) thi im 24 gi sau gy vim. Dch chit Ulva
reticulata khng lm gim vim cp tnh trn m hnh gy vim chn chut bng
carrgeenin (P > 0,05).
* Trn m hnh gy trn dch mng bng
Kt qu tc dng ca dch chit Sargassum swartzii (S) v Ulva reticulata
(U) ln th tch dch r vim, hm lng protein trong dch r vim v s lng
bch cu trong dch r vim c ch ra trn bng VII.2.9.b-d.
Bng VII.2.9.b. Tc dng ca dch chit Sargassum swartzii (S) v Ulva
reticulata (U) ln th tch dch r vim
L
i chng (ung NaCl 9,
1 ml/100 g)

Lng dch r vim (ml)

P so i chng

3,621,30

Indometacin 25 mg/kg

2,3 0,73

p < 0,05

Dch chit S (175mg/kg)

1,62 0,39

p < 0,01

Dch chit S (350mg/kg)

2,22 0,68

p < 0,01

Dch chit U (175mg/kg)

3,9 0,81

p > 0,05

Dch chit U (350mg/kg)

1,76 0,32

p < 0,01

Kt qu bng VII.2.9.b cho thy, dch chit t Sargassum swartzii liu

175mg/kg v liu 350mg/kg lm s lng dch r vim gim r rt so vi i
chng (P <0,05). Trong khi Ulva reticulata ch c tc dng liu 350mg/kg
(P <0,05).
522

Bng VII.2.9.c. Tc dng ca dch chit Sargassum swartzii (S) v Ulva

reticulata (U) ln hm lng protein trong dch r vim
L
i chng (ung NaCl 9,
1 ml/100 g)

Hm lng protein (mg/dl)

P so i chng

50,64 7,81

Indometacin 25 mg/kg

47,45 5,17

P > 0,05

Dch chit S (175mg/kg)

51,77 6,54

P > 0,05

Dch chit S (350mg/kg)

49,44 2,67

p > 0,05

Dch chit U (175mg/kg)

48,55 4,17

p > 0,05

Dch chit U (350mg/kg)

49,75 2,51

p > 0,05

Kt qu bng VII.2.9.c cho thy, dch chit Sargassum swartzii v Ulva

reticulata c 2 liu khng nh hng n hm lng protein trong dch r vim
so vi l chng (P >0,05).
Bng VII.2.9.d. Tc dng ca dch chit Sargassum swartzii (S) v Ulva
reticulata (U) ln s lng bch cu trong dch r vim
L
i chng (ung NaCl 9,
1ml/100 g)

S lng bch cu (g/L)

P so i chng

25,07 8,76

Indometacin 25 mg/kg

18,70 3,85

< 0,05

Dch chit S (175mg/kg)

17,73 4,57

< 0,05

Dch chit S (350mg/kg)

11,72 1,45

< 0,01

Dch chit U (175mg/kg)

13,67 3.15

< 0,01

Dch chit U (350mg/kg)

13,47 0,88

< 0,01

Kt qu bng VII.2.9.d cho thy, dch chit Sargassum swartzii v Ulva

reticulata c 2 liu 175 v 350mg/kg u c tc dng lm gim r rt s lng
bch cu trong dch r vim so vi l i chng (P <0,05). Tc dng gim s
lng bch cu trong dch chit t hai loi to nghin cu c xu hng tt hn
khi s dng Indometacin 25mg/kg.

523

VII.2.10. Tc dng chng vim mn

Tc dng ca dch chit t Sargassum swartzii (S) v Ulva reticulata (U)
ln trng lng u ht c ch ra trn bng VII.2.10.
Bng VII.2.10. Tc dng ca dch chit t Sargassum swartzii (S) v Ulva
reticulata (L) ln trng lng u ht
L
i chng (ung NaCl 9,
1ml/100 g)

Trng lng u (mg)

P so i chng

105,5 14,84

Prednisolon 5 mg/kg

86,67 7,35

< 0,05

Dch chit to S (175mg/kg)

84,25 12,89

< 0,05

Dch chit to S (350mg/kg)

72,0 18,09

< 0,01

Dch
chit
(175mg/kg)

to

105 23,18

> 0,01

Dch
chit
(350mg/kg)

to

108.4 20,53

> 0,01

Kt qu bng VII.2.10 cho thy, dch chit t Sargassum swartzii liu 175 v
350mg/kg c tc dng chng vim mn tnh thng qua lm gim trng lng khi u
ht so vi chng (P <0,05), tc dng ny c xu hng tt hn khi s dng
Prednisolon liu 5mg/kg. Dch chit Ulva reticulata khng c tc dng chng
vim mn (P >0,05).
VII.2.11. Hm lng lipit tng s
- Mu S (Cao Sargassum swartzii):
23,69% trng lng ti
- Mu U (Cao Ulva reticulata):
19,32% trng lng ti
Kt qu phn tch trn cho thy trong hai loi cao th nghim, mu cao
rong Sargassum swartzii cha hm lng lipit tng s cao hn 4,37% so vi mu
cao rong Ulva reticulata.
VII.2.12. Thnh phn axit bo
Thnh phn axit bo ca hai mu cao rong S v U c th hin bng
VII.2.12.a-b
Bng VII.2.12.a Thnh phn axit bo mu to S

Myristic

Thi
gian
lu (Rt)
11,92

Hm
lng
[%]
0,92

Axit 9 tetradecenoic

12,62

0,25

C15:0

Axit pentadecanoic

Oleic

12,94

0,35

C15:1(n-5)

Axit 10 pentadecenoic

13,76

0,72

TT

Axit bo

Tn khoa hc

Tn thng

C14:0

Axit tetradecanoic

C14:1(n-5)

3
4

524

C16:0

Axit hexadecanoic

Palmitic

16,76

53,19

C16:1(n-7)

Axit 9 hexadecenoic

17,64

5,73

C17:1(n-7)

Axit 10 heptadecenoic

20,84

0,49

C18:0

Axit octadecanoic

Stearic

23,19

0,85

C18:1(n-9)

Axit 11 octadecenoic

24,73

9,02

10

C18:2(n-6)

26,71

1,39

11

C18:3(n-6)

28,28

0,15

12

C18:3(n-3)

Axit 9,12 octadecenoic

Axit 6,9,12
octadecatrienoic
Axit octadecatrienoic

29,89

7,97

13

C20:0

Arachidic

31,65

4,43

14

C20:4(n-6)

Arachidonic

39,01

0,66

15

C22:0

Axit eicosanoic
Axit 5,8,11,14
eicosatetraenoic
Axit docosanoic

Behenic

41,68

0,76

16

C22:3(n-3)

49,69

0,27

17

C22:5(n-3)

Axit docosapentaenoic
Axit 7,10,13,16,19
docosapentaenoic

Clupanodonic

53,70

0,21

Others

12,85

Tng cc axit bo no

60,50

Tng cc axit bo khng no

26,65

Bng VII.2.12.b. Thnh phn axit bo mu to U

Myristic

Thi
gian
lu (Rt)
11,99

Hm
lng
[%]
2,77

Axit 9 tetradecenoic

12,66

0,34

C15:0

Axit pentadecanoic

Oleic

13,00

0,48

C15:1(n-5)

Axit 10 pentadecenoic

13,39

0,32

C16:0

Axit hexadecanoic

Palmitic

16,63

24,45

C16:1(n-7)

Axit 9 hexadecenoic

17,65

1,35

C17:1(n-7)

Axit 10 heptadecenoic

20,70

0,47

C18:0

Axit octadecanoic

Stearic

23,23

4,48

C18:1(n-9)

Axit 11 octadecenoic

24,45

31,87

10

C18:2(n-6)

26,80

22,69

11

C18:3(n-6)

28,51

0,68

12

C18:3(n-3)

Axit 9,12 octadecenoic

Axit 6,9,12
octadecatrienoic
Axit octadecatrienoic

29,87

2,79

TT

Axit bo

Tn khoa hc

Tn thng

C14:0

Axit tetradecanoic

C14:1(n-5)

525

13

C20:0

Axit eicosanoic

Arachidic

31,72

1,38

14

C20:1(n-9)

33,06

0,38

15

C20:4(n-6)

Arachidonic

38,82

1.81

16

C22:0

Axit 11 eicosenoic
Axit 5,8,11,14
eicosatetraenoic
Axit docosanoic

Behenic

41,68

0.62

17

C24:0

Axit tetracosanoic

Lignoceric

52,05

0.73

Others

2.39
Tng cc axit bo no

34,91

Tng cc axit bo khng no

62,70

Kt qu c ch ra trong bng VII.2.12.a-b cho chng ta thy:

- Tng cc axit bo no v tng cc acit bo khng no ca hai mu cao rong
nghin cu c s khc bit nhau r rt. mu S, tng cc acit bo no cao gp
2,27 ln so vi tng cc axit bo khng no (60,50% so vi 26,65%), trong khi
mu U th ngc li, tng cc axit bo khng no li cao gp 1,8 ln tng cc
axit bo no (62,70% so vi 34,91%). iu ny cng c ngha l tng cc axit bo
no ca mu S cao hn khong 1,73 ln so vi mu U (60,50% so vi 34,91%) cn
tng cc axit bo khng no th ngc li, mu U cao gp khong 2,35 ln so vi
mu S (62,70% so vi 26,65%).
- V thnh phn axit bo: Mu U c hm lng axit arachidonic (ARA,
C20:4(n-6)) cao gp 2,74 ln so vi mu S (1,81% so vi 0, 66%). ARA l mt
loi axit bo thuc h Omega-6, tin cht ca cc Eicosanoid gy vim. V vy,
kt qu phn tch ny c th l mt gi cho vic la chn i tng to bin cho
cc th nghim th hot tnh khng vim.
VII.2.13. Kt qu chit phn lp
T kt qu phn tch thnh phn axit bo vi hm lng ARA nh trn, kt
hp vi kt qu mt s th nghim sng lc s b hot tnh khng vim nh: cao
tng s ca c hai loi rong bin nu trn liu dng 500mg cao/kg u c tc
dng gim au trn c hai m hnh gy au bng axit axetic v mm nng; v tc
dng chng vim cp, cao tng s ca rong bin Sargassum swartzii liu 500mg
cao/kg c tc dng chng vim cp trn c hai m hnh gy ph chn chut v
m hnh trn dch mng bng, cn cao tng s ca rong bin Ulva reticulata ch
c tc dng trn m hnh gy trn dch mng bng; cao tng s ca rong bin
Sargassum swartzii c 2 liu dng (175 v 350mg/kg) u c tc dng chng
vim mn tnh. Trong khi cao tng s ca rong bin Ulva reticulata li khng
c tc dng chng vim mn. Do vy, trong hai loi cao c nghin cu, do
hn ch v thi gian v kinh ph, chng ti chn cao Sargassum swartzii (cao
S) tin hnh chit phn lp v tin ti tinh sch, xc nh bn cht mt s cht
c th c trong phn cao c hot tnh khng vim.
T 50 gam cao metanol tng s chit xut t bt to Sargassum swartzii,
tin hnh chit phn lp bng cc dung mi theo th t l n-hexan, diclomethane

526

v etyl-axetat. Sau khi ct ui dung mi thu c 3 loi cao tng ng vi hm

lng c th c ch ra trong bng VII.2.13.
Bng VII.2.13. Hm lng cc loi cao Sargassum swartzii sau khi chit phn
lp
Loi cao

TL cao (gam)

Hm lng % so vi cao tng s

MeOH

50

100

n-Hexan

37,43

74,86

Dichlomethane (CH2Cl2)

1,9

3,80

Ethyl axetat (EtOAC)

0,8

1,60

Nh vy trong 3 loi cao thu c sau khi chit phn lp, cao n-hexan
chim hm lng cao nht v cao hn rt nhiu so vi hai loi cao cn li
(74,86% so vi 3.80% v 1,60%). Kt qu ny c th cho php d on trong
thnh phn ha hc ca to Sargassum swartzii, hm lng cc cht mu (cc sc
t), cc cht bo v cc cht t phn cc ni chung l tng i ln.
VII.2.14. Hot tnh khng sinh ca ba loi cao tch t cao metanol tng s
ca to S
C 3 loi cao n-hexan, diclomethane v etyl-axetat u c th s b
hot tnh khng vi sinh vt nhm xc nh xem thnh phn c hot tnh khng
vim c ho tan trong dung mi no. Kt qu th hot tnh khng sinh ca 3
loi cao Sargassum swartzii sau khi chit phn lp c th hin bng VII.2.14.
Bng VII.2.14. Hot tnh khng sinh ca 3 loi cao Sargassum swartzii sau khi
chit phn lp
Tn chng vi sinh vt kim nh
STT

Tn mu

Ec
IC50
g/mL

Pa
IC50
g/mL

Bs
IC50
g/mL

Sa
IC50
g/mL

Ca
IC50
g/mL

1
H (cao Hexan)
>256
>256
>256
>256
>256
64
2
C (cao CH2Cl2)
>256
>256
>256
>256
3
E (cao EtOAC)
>256
>256
>256
>256
>256
K hiu: Ec: Escherichia coli; Pa: Pseudomonas aeruginosa; Bs: Bacillus subtillis;
Sa: Staphylococcus aureus; Ca: Candida albicans

Kt qu ch ra trn bng VII.2.14 cho thy trong 3 mu cao c th hot

tnh khng sinh ch c mu cao dichlomethane (K hiu C) c hot tnh khng
chng Staphylococcus aureus vi gi tr IC50 l 64g/mL. Gi tr ny cho thy
hot tnh khng sinh ca mu th l tng i tt. iu ny cho php nh hng
tip tc phn tch phn cao ny xc nh bn cht ha hc ca phn dch chit
c hot tnh khng vim. Hai loi cao cn li khng th hin hot tnh khng sinh
(hoc c hot tnh nhng khng ng k).

527

VII.2.15. Tinh sch cht

a/ Phn cao Dichlomethane:
Sau khi xc nh c cao Dichlomethane c hot tnh khng vi sinh
vt kim nh, phn cao ny s c dng tch cc phn on bng sc k ct
(SKC). Trc khi tch cc phn on bng SKC, tin hnh nh tnh cc lp cht
c trong phn cao Dichlomethane bng sc k bn mng (SKBM) kt hp vi cc
loi thuc hin thng dng pht hin cc vt cht (nh: cn axit (45-50%
H2SO4 trong ethanol) + t nhit cao, dragendorff , UV ). Th nghim cho
thy phn cao Dichlomethane (CH2Cl2) c th cha vt cht thuc nhm Alkaloid
hin mu vng cam c pht hin bi thuc hin mu c trng Dragendorff.
Sc k cng cho thy vt alkaloid cng c mt trong phn cao n-Hexan.
Tin hnh SKC phn cao Dichlomethane tch cc phn on, gp
chung cc phn on c sc k tng t nhau, chng ti thu c hai nhm
phn on chnh ng ch , k hiu l DC1 v DC2 .
- DC1 l nhm p/ cha vt alkaloid d on, c hin di UV bc
sng 254 nm, do hm lng tng i thp nn trong th nghim ny chng ti
cha kt tinh v cha lm sch. Qu trnh lm sch s c thc hin trong cc
th nghim sau.
- Nhm p/ DC2 kt tinh cho tinh th dng bt, mu trng, hm lng
khong 30 mg (trong tng s 1,558 gam cao Dichlomethane em tch, tc l hm
lng DC2 chim gn 2%) . Ra sch tinh th bng cc dung mi thch hp v
kim tra sch bng SKBM. Sc k cho thy vt cht DC2 hin r di UV
bc sng 254nm, c Rf = 0,55 trong h trin khai Chloroform : Metanol
(9,5:0,5).
Lm kh hon ton tinh th DC2 v ghi ph xc nh cu trc:
+ Ph khi cho peak M = 123
+ Ph 1H-NMR cho 2 singlet 3,4 v 3,6 l ca hai nhm CH3
ng xa nhau, 1 singlet 7,8 l ca CH trong vng thm v mt tn hiu
12,7 l ca H lin kt vi N.
+ Ph 13C cho bit hai tn hiu 28,5 v 30,2 l ca hai nhm CH3
lin kt vi vng thm. Tn hiu 106 l ca CH bc hai trong vng thm khng
b th, cn tn hiu 140, 149, 151 l ca C bc bn vng thm b th. Tn
hiu 156 l ca C trong nhm acetal CHO. T cc tn hiu ca ph MS, 1HNMR, 13C-NMR cho thy phn t c khi lng 123 gm cc nhm sau:
2 CH3 (30)
1 CH (13)
3 C (36)
1 NH (15)
1 CHO

OHC

H3C

(29)

Cng thc phn t: C7H9NO

D kin cng thc cu to ca phn t nh sau:

528

CH3

N1

H
3,5-Dimethyl-4-al-pyrol

b) Phn cao n-hexan

(Phn cao ny c tin hnh SKC vi mc ch xem c th tch c
cht no c hm lng ln hay khng v hm lng phn cao ny ln (37,43 g,
chim 74,86% trong tng s cao metanol em tch).
SKC 4 gam cao n-Hexan thu c 80 p/ (100 mL/p), cc h dung mi
tng dn phn cc. SKBM kim tra cc p/ v gp chung cc p/ c sc k
tng t nhau, nhn thy c 3 nhm p/ ng ch , k hiu l H1 (p/ t 21, ,
53), HII (p/ t 59, , 62 c mu nu) v HIII (p/ t 71, , 76 c mu vng
cam).
SKBM v kim tra vi cc thuc hin mu c trng, nhn thy HI cha
vt cht thuc nhm sterol (hin mu hng cnh sen sau khi t bng cn axit),
HII cha vt cht mu nu (cha xc nh) v HIII cha vt cht mu vng cam d
on l alkaloid. Vic tinh sch HI, HII v HIII s c thc hin tip trong cc th
nghim sau ty thuc vo kh nng kt tinh ca chng.
Tip tc xc nh bn cht ha hc ca cht thuc nhm Sterol trong HI
bng cch SKBM so snh vi cht chun. Kt qu xc nh c HI cha cht
22-Dehydroclerosterol (www.lipitbank.jp/cgi-bin/detail.cgi)
+ Danh php quc t:
(22E,24S)-24ethylcholesta-5,22,25-trien-3-ol
+ Tn thng dng:
22-dehydroclerosterol
+ Cng thc phn t:
C29H46O1
+ Trng lng phn t trung bnh:
410,675
+ Cng thc cu to:
(Hnh VII.2.15)
+ Nhit nng chy: axetat 146-150 C
+ D liu ph NMR: axetat 1H-NMR
(CDCl3) 3-CH=4.601 (m), 3-Ac=2.030 (s), 6CH=5.370 (br d), 18-CH3= 0.691(s), 19-CH3=1.012
(s), 21-CH3=1.011 (d), 22-CH=5.243 (dd), 23CH=5.171 (dd), 24-CH=2.420 (q), 26-CH3=1.646
(d), 27-CH3=4.694 (m), 29-CH3=0.833 (t).
+ D liu ph khi MS: axetat: 452[M+]
(0.3%), 409(0.2%), 392(100%), 377(3%), 363(3%),
282(5%), 255(32%), 253(12%), 228(3%), 213(7%).

Hnh VII.2.15. Cng thc cu to

22-Dehydroclerosterol

VII.2.16. Kt qu th hot tnh khng sinh v hot tnh chng oxi ha ca

DC2
Sau khi tinh sch v xc nh c bn cht ha hc ca DC2, chng ti
tin hnh phn tch hot tnh khng vi sinh vt v hot tnh chng oxi ha ca
cht ny ( dng tinh th).
a. Kt qu th hot tnh khng sinh
Kt qu th hot tnh khng sinh c trnh by bng VII.2.16.a.

529

Bng VII.2.16.a. Hot tnh khng sinh ca DC2 (3,5-Dimethyl-4-al-pyrol)

Tn chng vi sinh vt kim nh
Tn mu

Ec
IC50
g/mL

Pa
IC50
g/mL

Bs
IC50
g/mL

Sa
IC50
g/mL

Ca
IC50
g/mL

>128

>128

>128

>128

>128

DC2 (3,5-Dimethyl4-al-pyrol)

K hiu: Ec: Escherichia coli; Pa: Pseudomonas aeruginosa; Bs: Bacillus subtillis;
Sa: Staphylococcus aureus; Ca: Candida albicans

Kt qu ch ra bng VII.2.16.a cho thy cht DC2 sau khi c lm sch

li khng th hin hot tnh khng cc chng vi sinh vt kim nh nng <
128 g/mL. Nh vy, so vi kt qu th hot tnh khng sinh ca 3 loi cao
Hexan, CH2Cl2, EtOAC (ch ra bng 17), trong cao CH2Cl2 c hot tnh
khng sinh vi IC50 l 64 g/mL th thy rng: mc d cht DC2 nm trong phn
cao Dichlomethane CH2Cl2 nhng khi tch ra dng tinh sch th thy khng cn
hot tnh. iu ny cho thy trong qu trnh tinh sch cht DC2 c th lm mt i
hot tnh khng sinh ca n. iu ny cn phi kim tra li c th c kt lun
cui cng.
b. Kt qu th hot tnh chng oxi ha
Kt qu th hot tnh chng oxy ho c th hin bng VII.2.16.b.
Bng VII.2.16.b. Hot tnh chng oxi ha ca DC2 (3,5-Dimethyl-4-al-pyrol)
STT

K hiu mu

Nng mu
(g/mL)

SC%

Kt qu

1
2

Chng (+)
Chng (-)

50
-

66,8 0,0
0,00 0,00

Dng tnh
m tnh

DC2

2,1 0,00

m tnh

Kt qu bng VII.2.16.b cho thy, gi tr SC% ca cht DC2 ch l 2,1%.

Trong khi , mt mu th c coi l c hot tnh chng oxi ha khi gi tr SC%
50%. Theo , c th kt lun rng cht DC2 khng th hin hot tnh chng
oxi ha trn h DPPH.
* C th l gii hin tng cht DC2 dng tinh khit khng c hot tnh
ca bi mt s nguyn nhn sau:
- Hot tnh khng sinh ca phn cao CH2Cl2 c th khng phi ch do mt
cht nht nh gy ra m l do tc ng tng tc ca mt s cht cng c mt
trong phn cao ny gy ra. V vy khi tch ring ra, cht DC2 dng tinh sch
khng th hin hot tnh.
- Nguyn nhn th hai c th l do tc ng ca qu trnh tinh sch. Trong
qu trnh lm sch DC2, do nh hng ca dung mi (cha cc cht bo qun...)
nn tinh th b bm cc cht c ngun gc t dung mi, cc cht ny lm nhiu
ph ca DC2 nn chng ti tip tc lm sch loi b cc cht bm ny. C

530

th do qu trnh ra sch tinh th ko di v tri qua cc iu kin khc nhau (nh

nhit , hn hp dung mi...) nn hot tnh ca cht DC2 gim i ng k. V
vy, nng < 128 g/mL, cht DC2 khng cn th hin hot tnh khng sinh
v trn h DPPH, cht DC2 khng th hin hot tnh chng oxi ha.
- Nguyn nhn th ba c th l do hot tnh khng sinh ca phn cao
CH2Cl2 l do cc cht khc (m chng ti cha tin hnh tinh sch tip) ch
khng phi do cht DC2. Khi , cht DC2 hon ton khng c hot tnh khng
sinh.
Nh vy, qu trnh tinh sch cht DC2 c th lm mt i hot tnh khng
sinh ca n. iu ny cn phi kim tra li c th c kt lun cui cng.
VII.3. Kt lun
- thu thp v nh tn c 19 loi rong bin Vit Nam phn b cc tnh
min Bc, Trung v Nam ca Vit Nam.
- xy dng c quy trnh tch dch chit c hot tnh khng vim t cc loi
rong bin Vit Nam bng dung mi metanol; phng php sng lc nhanh cc
loi rong to bin trong khun kh ti c hot tnh khng vim bng dung
mi metanol, iu kin ti u cho vic sng lc cht khng vim t cc loi
rong bin Vit Nam l 1 gi vi nng 0,08mg/l dch chit khng vim
c xc nh.
- Trn c s ca phng php ny, trong tng s 19 loi rong bin c sng lc
th Sargassum swartzii v Ulva reticulata c hot tnh khng vim cao v c
tim nng khai thc c pht hin;
- thu c 300g v 200g dch chit c c c hot tnh khng vim cao t
Ulva reticulata v Sargassum swartzii, tng ng tng loi mt;
- Kt qu th hot tnh khng sinh cho thy: Ulva reticulata c hot tnh khng
Bacillus subtilis vi gi tr IC50 = 62,15 g/ml v Sargassum swartzii c hot
tnh khng Staphylococcus aureus vi gi tr IC50 = 56,53 g/ml.
- tin hnh th hot tnh khng vim ca cc dch chit thu c t Ulva
reticulata v Sargassum swartzii trn ng vt thc nghim (th c tnh cp,
tc dng gim au bng axit axetic, gim au mm nng, gim vim mn bng
u ht, gy vim cp mng bng bng carageenane):
+ Dch chit t Sargassum swartzii v Ulva reticulata ung n liu
66g/kg cha c chut cht. Thuc t c tnh cp, cha tnh c LD50 theo
ng ung.
+ Dch chit Sargassum swartzii v Ulva reticulata liu 500mg/kg c tc
dng gim au trn c hai m hnh gy au bng axit axtic v mm nng.
+ Dch chit Sargassum swartzii liu 500mg/kg c tc dng chng vim
cp trn m hnh gy ph chn chut v m hnh trn dch mng bng. Dch
chit Ulva reticulata ch c tc dng chng vim cp trn m hnh gy trn
dch mng bng.
+ Dch chit Sargassum swartzii liu 175 v 350mg/kg c tc dng chng
vim mn tnh. Dch chit t Ulva reticulata khng c tc dng chng vim
mn tnh.
531

- xc nh c hm lng lipit tng s ca hai mu cao rong Sargassum

swartzii v Ulva reticulata theo th t l 23,69% v 19,32% trng lng ti.
- Thnh phn axit bo ca hai mu cao rong Sargassum swartzii v Ulva
reticulata c xc nh. Trong , hm lng axit arachidonic (ARA C20:4(n-6)) ca cao Ulva reticulata l 1,81%, cao gp 2,74 ln so vi hm
lng axit Arachidonic ca mu cao Sargassum swartzii (0,66%). Tng cc
axit bo no v axit bo khng no ca hai mu cao ny ln lt l 60,50% v
26,65%; 34,91% v 62,70%.
- T cao metanol tng s ca Sargassum swartzii sau khi chit phn lp thu c
ba loi cao vi hm lng tng ng nh sau: cao n-Hexan (37,43 g) chim
74,86%, cao Dichlomethane (1,9g) chim 3,80% v cao Ethyl axetat (0,8 g)
chim 1,60%.
- Trong ba loi cao n-hexan, dichlomethane v ethyl axetat th ch c cao
dichlomethane c hot tnh khng Staphylococcus aureus vi gi tr IC50 l
64g/mL, hai loi cao cn li khng th hin hot tnh khng sinh (hoc c
hot tnh nhng khng ng k).
- Phn tch ha hc phn cao dichlomethane v cao n-hexan cho thy c hai phn
cao ny u cha cht thuc nhm alkaloid. Trong , phn cao dichlomethane
c hai cht quan tm, k hiu l DC1 v DC2. Cht d on l alkaloid trong
phn cao dichlomethane c k hiu l DC1, cn DC2 tinh sch c l mt
cht kt tinh dng bt mu trng, hm lng 30 mg. Phn tch cc d liu ph,
chng ti xc nh c bn cht ha hc ca DC2 l 3,5-Dimethyl-4-alpyrol, c cng thc phn t l C7H9NO.
- Phn tch ha hc phn cao n-Hexan cho thy trong phn cao ny c 3 cht
quan tm, k hiu l HI, HII v HIII, trong HI xc nh c l 22Dehydroclerosterol, HIII l cht d on thuc nhm Alkaloid (cha xc nh
c th) cn HII cha xc nh.
- Kt qu th hot tnh khng sinh v hot tnh chng oxi ha cho thy, cht DC2
khng th hin hot tnh khng cc chng vi sinh vt kim nh nng <
128 g/mL v khng th hin hot tnh chng oxi ha trn h DPPH (1,1diphenyl-2-picrylhydrazyl).

532

Chng VIII. Nghin cu thm d kh nng sinh cc

cht c hot tnh sinh hc t vi sinh vt bin.
VIII.1. M u
Vi sinh vt bin tr thnh mt vn quan trng trong vic nghin cu cc
sn phm vi sinh vt mi trong th k 21. Nhiu hp cht th cp c hot tnh YSinh hc nh cc enzim, cc protein, cc cht chng ung th, chng virus, chng
vim nhim c bit m cc ngun khc khng c c tch chit t sinh vt
bin (hot tnh ca chng thng cao gp hn 100 ln so vi sinh vt trn cn)
[299].
Cc cht c HTSH t vi sinh vt bin c th c chia thnh 6 nhm cht sau:
1. Cc cht c hot tnh khng sinh (khng khun, khng nm, khng vim
nhim, dit cn trng...)
2. Cc cht dit virus.
3. Cc cht gy c t bo v chng ung th.
4. Cc cht c ch enzim.
5. Cc cht chng oxy ho.
6. Cc vitamin, carotenoid v cc axit bo khng no mch di.
Cho n nay c hn 1.200 hp cht c tch t VSV bin, trong c
nhng cht c nht v nh m cc sinh vt trn cn khng c c. Bin l mi
trng sng rt c bit, v ngho cht dinh dng v mn cao. Vi sinh
vt bin c kh nng thch nghi vi cc iu kin mi trng bin lun b thay
i, iu ny m ra cc trin vng sn xut cc hp cht hu c th cp c o.
T dng vi khun di bin su, ngi ta tch c mt hp cht Macrolid
mi gy c t bo. Khi nui cy, chng sn sinh ra mt lot hp cht thin nhin
mi m trn cn cha tng c nh Macrolactin A, c tc dng c ch t bo ung
th, bo v t bo limpho T, chng HIV [299]. Tuy nhin, nui cy v phn lp
cc vi sinh vt bin l cng vic rt kh khn. Nguyn nhn vic kh nui cy l
do bin khi ngho cht dinh dng, c mui khc nhau, vi sinh vt bin
thch nghi vi vic i v mi trng ngho nn khng pht trin trn mi trng
giu cht dinh dng hn. Ngi ta s dng cc mi trng v cc phng
php khc nhau tch cc nhm vi sinh vt bin khc nhau.
S lng vi sinh vt bin khng cao nh trn cn. Mt s kt qu cho
rng, nc bin cha khong 10 - 104 CFU /ml v c th bt gp chng cc iu
kin sng khc nhau ln hn so vi cc vi sinh vt trn cn. Vi sinh vt bin c
th phn lp c t trm tch bin, nc bin, t cc i tng hot ng hoc
bt ng m vi sinh vt c th cng sinh.
Hiu c qui lut phn b ca vi sinh vt bin s lm d dng v to iu
kin thun li thu thp chng t cc vt mu thch hp cho vic phn lp nh
hng cc nhm vi sinh vt c hot tnh sinh hc c phn loi.
Vi sinh vt cng sinh vi thc vt v ng vt khng xng sng c
ngha rt ln trong vic tch chit cc cht th cp c hot tnh sinh hc. S cng
sinh gia vi khun v bt bin l mt loi cng sinh ph bin nht [300]. Mt
trong nhng loi vi khun cng sinh vi bt bin l Rhopaloeides odorabile.

533

Ngoi ra rt nhiu loi vi khun l thng v cha c m t thng cng sinh

vi to bin. Khong 30% vi khun cng sinh vi to xanh (Hamimeda sp.) v to
nu (Lopophora sp.) c th nui cy c l vi khun Gram dng (+). Cc vi
khun Gram dng (+) thng gp trm tch bin v chng c kh nng sinh
cc cht th cp c hot tnh dit cn trng, cc enzyme gy c cho t bo
[301]. T nm si cng sinh vi to nu bin Roseuvuigea cng tch c cht
khng sinh mi Pestanlone c ch sinh trng vi khun [302]. Cc cht chng
oxy ho Superoxidase glutathion (SOD) v mt s dn xut mi Hydroquinone,
Oxepinamides A-C, Fumiquinazolines H-1 cng c tch chit t vi khun bin
v nm si bin Acremonium spp. [303]. X khun bin cng l ngun tch chit
cc cht c HTSH mi m cc ngun trn cn khng c [299, 304]. Ngoi cc
cht khng sinh c tch t nhm x khun Streptomyces, Diazepinomicin- mt
hp cht alkaloid mi khng khun cng c tch t x khun bin
Micromonospora.
Trn th gii, vic nghin cu v ng dng cc cht hot tnh t vi sinh vt
bin c tin hnh trong nhiu nm qua trn qui m rng ln v thu c
nhiu kt qu ng k [303-307]. Vi nm gn y VN c mt s cng trnh
v iu tra s lng, s phn b v tnh a dang ca mt s nhm VSV bin nh
nm men bin, vi khun phn gii cht hu c [308-310]. T cc kt qu ny, cc
nhm nghin cu ch to ra c mt s ch phm sinh hc cha cc chng vi
sinh vt sng hu ch lm sch mi trng nc nui tm v thc n b sung
cho nui trng thu sn. Tuy nhin, cc nghin cu v cc cht c HTSH t VSV
bin cn l mt mng trng.
Vit Nam c b bin di trn 3000 km, tnh a dng vi sinh vt rt phong
ph. V vy, vic nghin cu cc cht hot tnh sinh hc t vi sinh vt bin rt c
trin vng, m rng vic s dng ngun ti nguyn sinh vt bin mt cch c
hiu qu. l mt hng nghin cu mi, cn thit, c ngha khoa hc v ng
dng.
VIII.2. KT QU v tho lun
VIII.2.1 Phn lp v xc nh s lng ca cc nhm vi sinh vt bin trong
nc bin, trm tch bin v trong cc mu rong to, c bin v s huyt
Vit Nam
60

12

52.5

50

10

40

8.5

30

24

20
10

10

14.9

12.7

9.1

8.6

6.9

4.4

S lng VSV x1000 (CFU/g)

% vi khun

% x khun

T l % cc nhm VSV

% nm men

% nm si

T l % chng c KN khng VSVK

Trm tch

T l % chng c KN i
khng VSVK
Nc bin

Hnh VIII.2.1.a. T l % v kh nng khng VSV kim nh ca cc nhm VSV phn lp

c

534

T 81 mu vt bin, chng ti phn lp c 1052 chng VSV, bao

gm 552 chng vi khun (chim 52,5% tng s chng), 253 chng nm si
(chim 24%), 157 chng x khun (14,9%) v 90 chng nm men (8,6%) (hnh
VIII.2.1.a). Trong cc mu trm tch, s lng VSV dao ng t 0 n 5,6 . 104
CFU/ml, cao hn so vi s lng VSV trong cc mu nc bin vi chc ln (0 7,6 . 103 CFU/ml). Trong tt c cc mu th s lng vi khun t cao nht (101
n 104), tip n l nm si, x khun, cn nhm nm men xut hin t nht (t
0 101 CFU/ml).
Hnh VIII.2.1.b cho thy, s lng VSV trong cc mu trm tch v nc
bin ph thuc vo mn, khong cch xa b, su. S lng VSV trong cc
mu trm tch dao ng t 0 n 5,6.104CFU/g nhiu hn s lng vi sinh vt
trong cc mu nc bin (0-7,6.103CFU/ml). Trong tt c cc mu th s lng vi
khun t cao nht (10-104), tip n l nm si, x khun, cn nhm mn men
xut hin t nht. Trong , cc mu nc bin c mn 3% s lng vi
khun xut hin t hn vi s lng 102CFU/ml, ring mu nc bin Trng Sa
s lng vi khun l 7,6.103CFU/ml cao hn cc vng bin Ph Yn v Bnh
nh. Hu ht mu trm tch bin c mn 2% Bnh nh v Qung Bnh, s
lng vi khun dao ng 104CFU/g, ln hn s lng vi khunt mu nc
bin 10-100 ln (102-103CFU/ml) v cng cao hn 100 ln so vi s lng vi
khun trong cc mu san h v rong trng bin Trng Sa, Nha Trang
(102CFU/ml).
Nm men, x khun v nm si trong cc mu nc bin s lng rt thp
t 0 n 101CFU/ml, thp hn so vi hu ht cc mu trm tch bin Bnh nh,
Quang Bnh (102-103CFU/g). 27 mu phn tch, nm men ch xut hin 7/27
mu, x khun c 13/27 mu v hu ht vi s lng thp 10-102CFU/ml(g).
Trong khi , vi khun v nm si u xut hin tt c cc mu phn tch ngoi
tr 2 mu trm tch Qung Bnh l khng c mt nm si.

10

10

10

8 .5
8

S lng VSV (CFU/ml)

T l % s chng VSV khng VSVK (%)

10

7 .6

5 .5

T l % s chng VSV khng

VSVK (%)

10

11

12

60
55

10.5

10.6
10.3

10.3

50
40

10

30
9.5

9.3

20
10

10
10

Lng VSV (CFU/g)

12

5.5
0

8.5
<10

80-100

350-500

10001200

Khong cch xa b (m)

0

0
1 - 1 ,5

2 ,5 - 3

Kh? nng K

m n (% )
K h nng K

Slg VSV X1000 (CFU/ml)

S lg V S V x 1 0 0 (C F U / m l)

Hnh VIII.2.1.b. nh hng ca mn v khong cch xa b ln s lng v kh

nng khng VSV kim nh ca vi sinh vt bin Vit Nam
Khc hn so vi cc mu nc v trm tch gn b, cc mu thc vt
(rong trng, to) v ng vt bin (san h, ngao s huyt) vi khun xut hin rt
535

t, cao nht l 102 CFU/g, cn nm men v x khun hu nh khng c (bng

VIII.2.1). Theo mt s ti liu, cc chng vi khun cng sinh vi ng vt bin
thng sinh cc cht c hot tnh sinh hc c bit. T c hi i Ty Dng
phn lp c chng Acromonas salamoniada, t c ui vng Yeloowtail
Sereola quinqueradiate phn lp c cc chng vi khun Streptococcus spp. v
t tm hm Penaeus monodon phn lp uc vi khun Vibrio hareyi, Vibrio
splendidus. Rut c bin c h vi khun rt phong ph, trong c nhiu chng
khng c vi khun gram (+) [307, 311], Tuy nhin, trong s 103 chng VSV
phn lp c t rong to bin, c, s huyt ca Vit Nam (bng VIII.2.1), khi
th kh nng khng khun th hu nh chng khng c hot tnh.
Cc khun lc VSV bin rt a dng v phong ph v hnh thi, kch
thc. iu ny ha hn s a dng ca cc hp cht c HTSH m chng c th
sinh ra. T cc kt qu phn tch chng ti nhn thy, s lng VSV trong cc
mu ph thuc vo mn, khong cch xa b v su ca tng ly mu. S
lng ca cc nhm VSV nc bin c mn 3% t hn t 10 100 ln so vi
s lng VSV trm tch bin c mn 2%. Cc mu trm tch bin, nc bin
gn b v nc b mt do cha nhiu cht hu c nn d phn lp v nui cy
VSV bin hn cc mu xa b. Kt qu ny ph hp vi nghin cu ca Li
Thy Hin [309], ng Cm H [308], Tng Kim Thun [312].
Bng VIII.2.1. S lng VSV cng sinh trong cc mu ng vt v thc vt bin
Mu

a im

Nm si

Nm men

X khun

Vi khun

San h

Nha Trang,
Trng sa

Cc loi c

Ch H Ni

15

74

Ngao, s

Hi Phng

20

Rong trng

Nha Trang

Rong

Nha Trang

19

103

Tng s

VII.3.2 Sng lc s b, th hot tnh sinh hc ca cc chng VSV bin phn

lp uc (khng sinh, khng nm, gy c t bo, sinh enzim v axit bo
khng bo ho..)
Cc chng VSV bin phn lp c th hot tnh khng khun, khng
nm. Kt qu trnh by trong bng VIII.2.2.a cho thy, trong s 1052 chng VSV
bin c th kh nng i khng th khng c bt k chng no i khng c
vi vi khun kim nh Gram (-) E.coli v P.aeruginose. C 38 chng vi khun
(chim 3,6% s chng), 20 chng x khun (1,9%), 23 chng nm si (2,1%) v
4 chng nm men (0,38%) khng vi khun Gram (+) B.subtilis vi ng knh
vng v khun dao ng t 7- 46mm, c 1 chng nm men v 1 chng x khun
khng c vi khun gram (+) S.aureus vi hot tnh trung bnh, ng knh vng
v khun t 14-15mm. c bit c 1 chng x khun khng c nm men gy
bnh C.albicans.
536

Bng VIII.2.2.a. Kh nng khng 5 chng VSV kim nh ca 1052 chng VSV
bin
Nhm
vsv

B.subtilis
E.coli
K d-d1
K
d-d1
(mm)
(mm)
552 38 7-18
0
253 23 10-46 0
157 20 18-40 0
90
4
8
0
1052 85

TTC

VK
NS
XK
NM
Tng s

S.aureus
P.aeruginose C.albican
K
d-d1
K
d-d1
K d-d1
(mm)
(mm)
(mm)
0
0
0
0
0
0
+
14
0
+ 5-12
+
15
0
0

TTC: s chng th nghim, VK: vi khun, NS: nm si, XK: x khun, NM: nm men, K: khng
VSV kim nh; d1-d2: ng knh vng v khun (mm).

T bng VIII.2.2.b cho thy, trong 4 nhm VSV bin phn lp c, th

nhm x khun c t l s chng c hot tnh khng khun, khng nm cao nht
(12,7%), tip n l nhm nm si (9,1%). Kh nhiu chng x khun c ng
knh vng khng khun ti 40mm, vng khng nm ti 24mm. Nhm vi khun v
nm men hu nh khng c kh nng khng khun, khng nm, hoc c th hot
tnh rt thp. ng knh vng khng khun, khng nm u di 10mm. c
bit, trong qu trnh phn lp tuyn chn tuyn c mt chng x khun TG1
c kh nng khng chng nm si 55 cng phn lp t Ph Tam Giang, vng
khng 18mm (hnh VIII.2.2).

x khun khng mc ng knh

vng khng 18mm

Hnh VIII.2.2. X khun TG1 khng chng nm si 55 Tam Giang

Bng VIII.2.2.b. Kh nng khng khun, khng nm ca 4 nhm VSV bin
Nhm VSV
bin
Vi khun
Nm si
X khun
Nm men
VSV bin

Kh nng khng khun

Schng
T l (%)
c HT
38/552
6,8
23/253
9,1
20/157
12,7
4/90
4,4
85/1052
8,1

D1-D2
7-18
10-46
18-40
7
7-46

537

Kh nng khng nm gy bnh

Schng
T l (%)
D1-D2
c HT
0/552
0
0
0/253
0
0
2/157
1,3
12-24
0/90
0
0
2/1052
0,2
12-24

Theo mt s ti liu [305, 307, 311], th nhiu chng VSV bin c kh

nng tng hp mt s c t gy cht cn trng. Chng ti th hot tnh gy
c t bo trn giun t v kh nng dit su t v su xanh ca 142 chng VSV
bin. Kt qu c trnh by trn bng VIII.2.2.c.
Bng VIII.2.2.c. Kh nng dit su v giun t ca VSV bin phn lp c
Nhm
VSV bin

Kh nng dit su t, su xanh

S chng c hot
T l %
tnh/tng s chng
8/80
10
0/30
0
0/20
0
0/12
0
8/142
5,7

Vi khun
Nm si
X khun
Nm men
VSV bin

Kh nng gy c t bo
S chng c hot
T l %
tnh/tng s chng
2/80
2,5
4/30
13
1/20
5
0/12
0
7/142
5,1

T bng VIII.2.2.c cho thy, c 10% s chng VSV (8/80) c kh nng

dit su hi v u thuc nhm vi khun gram (+) v hu ht c phn lp t
trm tch bin. iu ny cng ph hp vi nhn nh ca mt s tc gi [307,
311], rng t trm tch bin su thng phn lp c cc chng vi khun gram
(+) c kh nng sinh c t dit su. Cc chng nm men bin khng c hot tnh
dit su v gy c t bo. Nm si v x khun ch c kh nng gy c t bo,
khng c kh nng dit su. Cn nhm vi khun th c c 2 kh nng trn. T l
cc chng vi khun c kh nng dit su cao nht (10%) v gy c t bo ch
chim 2,5%. T l chng nm si c kh nng gy c t bo kh cao, ti 13%,
cn x khun ch c 5% tng s chng c kh nng ny.
Trong s 1052 chng VSV bin c th kh nng i khng vi 5 loi
VSV kim nh, c 3 chng c vng v khun ln nht. 3 chng ny u phn lp
c t trm tch bin. Chng nm si T10.6 c vng khng vi khun gram (+)
B.subtilis l 39-46mm; chng x khun VN2 c ng knh vng v khun khng
2 loi vi khun gram (+) B.subtilis l 38mm v S.aureus l 24mm; chng vi
khun T9.5 c vng v khun khng B.subtilis l 18mm. C 3 chng ny c
kim tra kh nng gy cht t bo trn i tng l giun t. Kt qu c trnh
by trn bng VIII.2.2.d
Bng VIII.2.2.d. Kh nng gy cht giun t Derodigitata Miiler ca dch nui cy
03 chng VSV bin sau 45 pht (t l % giun cht )
Liu
lng
dch ml

Chng nm si T10.6

Chng x khun VN2

Chng vi khun T9.5

TN

TN

TN

0,5

100

30

95

40

100

30

1,0

96

45

95

48

96

40

1,5

94

60

92

72

96

62

Dch nui cy 3 chng VSV bin c ly tm, loi b cn v sinh khi.

iu chnh pH dch nui cy n 7,0. Tin hnh th nghim theo phng php
c m t trong phn phng php nu trn.
Kt qu bng VIII.2.2.d cho thy, dch nui cy 03 chng x khun VN2,
nm si T10.6 v vi khun T9.5 u c hot tnh gy c cho giun t
538

Derodigitata miiler - thc n cho c cnh. Sau 45 pht vi liu lng dch nui
cy l 1,5ml c 60-72% s giun t cht. Chng x khun VN2 c kh nng
gy cht giun t cao nht, t 72%. y ch l cc nghin cu thm d nn tc
nhn gy cht giun t chng ti cha c r. Cn nghin cu su hn.
Nh trn bng VIII.2.2.c trnh by, cc chng x khun, nm si v
nm men bin khng c kh nng dit su. Ch c mt s chng vi khun bin c
kh nng ny. Trong s 3 chng vi khun c th nghim hot tnh dit su t
v su xanh th chng vi khun T 9.5 c hot tnh dit su t tt nht, sau 5 ngy
vi lng dch nui cy l 2,0ml c 80% s su cht (bng VIII.2.2.e).
Bng VIII.2.2.e. Kh nng dit su t, su xanh ca dch nui cy 03 chng vi
khun bin sau 5 ngy b ly nhim (t l % su cht)
Liu
Su xanh ( Heliothis armigera)
lng Chng
Chng
Chng
dch T3.4
T 9.5
T6.2
th
(ml) C TN C TN C TN

Su t (Plutella xylostella)
Chng
Chng
Chng
T3.4
T9.5
T6.2
C

TN

TN

TN

1,0

100

20

100

50

100

36

100

50

96

60

100

25

1,5

100

34

100

58

100

42

96

52

98

68

94

36

2,0

96

40

98

62

96

50

94

60

98

80

96

46

T l su cht c tnh theo cng thc Abbott C-T.100%, trong , C: s su sng l C, T:

s su cht l TN

C 3 chng vi khun trn c phn lp t trm tch bin v u l vi

khun Gram (+), c hnh thi khun lc ging chi Bacillus. So vi cc chng
VSV dit su phn lp t cc con su cht t nhin trn cnh ng u tng Ho
Bnh th kh nng dit su ca 3 chng vi khun ny khng cao bng, tuy nhin
chng cng c trin vng ng dng nh cc tc nhn sinh hc trong phng tr
su bnh khng gy nhim mi sinh nu c nghin cu su hn.
Bng VIII.2.2.f. Kh nng sinh enzym ca mt s chng VSV bin phn lp c

TT

1
2
3
4

Nhm VSV
bin

Vi khun
X khun
Nm si
Nm men
TSVSV

Sinh amylaza
S
Vng
chng
phn gii
(mm)
c hot
tnh
15/30
13-29
4/20
5-14
16/22
12-22
5/10
8-12
40/82

Sinh xenlulaza
S
Vng
chng
phn gii
(mm)
c hot
tnh
20/30
14-30
15/20
4-36
12/22
10-20
3/10
2-10
50/82

Ghi ch: HT: hot tnh; TS: tng s chng.

539

Sinh proteaza
S
Vng
chng
phn
c hot
gii
tnh
(mm)
6/30
12-26
2/20
8-13
1/22
5-10
3/10
7-10
12/82

T bng VIII.2.2.f cho thy, trong 82 chng c th nghim c trn 50%

s chng c hot tnh sinh amilaza v xenlulaza, ch c khong 12% s chng c
hot tnh proteaza. Nhm vi khun c hot tnh sinh 3 loi enzim cao nht (vng
phn gii xenlulaza ti 30mm), sau n nhm x khun (vng phn gii
xenlulaza ti 36mm) v nm si. Cn nhm nm men c hot tnh enzim thp
nht (vng phn gii ch khong 10mm).
Trong mu nc bin Trng Sa v Nha Trang, chng ti tch c 6
chng nm men bin v xc nh hm lng axit bo tng s. Kt qu c trnh
by bng8.
Axit bo cha no mch di c quan tm nhiu nht l axit bo loi -3
v -6. Chng l nhng axit bo cn thit cho sc khe, cho sinh trng v pht
trin.Thnh phn axit bo ca mt s chng nm men Candida c cc tc
gi nghin cu trong mt s bi bo [310]. Tuy nhin, hm lng axit bo -3 v
-6 ca cc chng ny cn tng i thp. Do , vic tm ra nhng loi nm
men c kh nng sinh ra cc axit bo cn thit -3 v -6 c hm lng cao
ng dng lm nguyn liu thuc v thc n b sung cho nui trng thy sn.
Bng VIII.2.2.g: Thnh phn axit bo ca 6 chng nm men phn lp t nc
bin trn mi trng Hansen dch th sau 24 gi (% ca axit bo tng s)
Chng
VSV

Axit bo TS
(%)/
trng 16:0
lng kh

16:1

18:0

18:1

18:2

18:3

NT1

4,73

16,11

5,00

4,18

33,76

28,18

5,79

NT2

23,63

9,76

2,60

24,29

12,00

6,55

NT3

4,91

17,26

4,52

32,24

28,48

5,72

NT8

16,35

18,93

2,84

38,26

38,55

6,42

TS1

5,25

13,04

4,91

3,60

50,85

18,78

8.83

TS2

8,29

11,23

1,32

7,99

57,40

12,30

7,63

4,76

T bng VIII.2.2.g cho thy, i vi tt c 6 chng nghin cu, axit bo

ch yu trong 2 phn on trung ho v c cc l Linolic (18:2), axit linolenic
(18:3) v axit palmitic(16:0), chim ti hn 80% axit bo tng s. C 6 chng
u khng sinh DHA (22:6) v ch c chng NT2 sinh EPA (20:5) vi hm lng
rt thp 0,13% tng s axit bo. Chng NT1, NT3 v NT8 sinh axit linoleic
(18:2) cao nht, t 28,48-38,55%, cn chng NT2, TS2 v TS1 sinh axit linolenic
(18;3) kh cao, t khong 6,55-8,83% axit bo tng s. So snh vi kt qu phn
tch thnh phn axit bo ca mt s chng nm men, x khun v nm si ca
mt s tc gi [310], cho thy hm lng axit bo khng bo ho ca 2 chng
NT1 v NT8 vo loi kh.
Nh vy, trong 6 chng th c th tuyn chn 2 chng NT1 v NT8
nghin cu qui trnh sn xut sinh khi nm men lm thc n cho c.

540

Bng VIII.2.2.h. Kh nng sinh trng v sinh tng hp astaxanthin ca 4 chng

nm men bin sinh sc t trn mi trng Hansen dch th
K hiu chng

Sinh khi kh
(g/l)

Carotenoid TS (g/g)

Astaxanthin (g/g)

NT5

9,2

450

240

NT8

8,4

194

120

TS5

8,2

246

HP4

7,4

201

Kt qu t bng VIII.2.2.h cho thy, 2 chng nm men NT5 v NT8 u c

kh nng sinh astaxanthin, tuy nhin hm lng astaxanthin trong sinh khi
khng cao (240 v 120 g/g).
Tm li, kt qu sng lc s b hot tnh sinh cht khng khun, khng
nm, sinh axit bo khng no, sc t carotenoit v enzim ngoi bo ca 1052
chng VSV bin phn lp c, chng ti tuyn chn c 11 chng c hot
tnh kh cao, trong c 3 chng x khun khng vi khun Gram(+), khng nm
v gy c t bo (VN1, VN2, VN3), 2 chng nm men sinh axit bo mch di
khng bo ho (TS1, NT8), 1 chng nm men sinh astaxanthin (NT5); 1 chng vi
khun Gram (+) sinh c t dit cn trng (T9.5) v 4 chng nm si khng vi
khun Gram (+) (c bit chng T10.6 c hot tnh rt cao 46 mm).
VIII.2.3. Quy trnh tuyn chn, sng lc cc VSV bin sinh cc cht c
HTSH
T 1052 chng VSV phn lp t cc mu nc, trm tch ng vt, thc
vt bin chng ti s dng sng lc cc chng c hot tnh sinh cao (ng
knh vng khng >22mm). Cc chng VSV bin c tin hnh sng lc theo cc
bc sau :
- Cc chng VSV bin c hot ha trn mi trng thch 1 ngy i vi vi
khun, 2 ngy i vi nm men, 7 ngy vi x khun v nm mc.
- Cc chng VSV bin hot ha c cy im trn cc a thch cha cc
VSV kim nh khc nhau gm : B.subtilis, S.aureus, E.coli, P.aeruginosa,
C.albicans.
- Nui cy trong t m cc nhit v thi gian khc nhau ty tng c im
sinh trng ca sau chng VSV kim nh. Sau , ly a thch ra quan st,
chng vi khun bin no to vng i khng vi VSV kim nh s c gi li
tip tc nghin cu (hnh VIII.2.3.a).
- Cc chng VSV bin c hot tnh c nui cy trn mi trng dch th (MPA
i vi vi khun, Hansen i vi nm men, Gause i vi x khun, Czapek i
vi nm si), trong 1 ngy i vi vi khun, 2 ngy i vi nm men, 7 ngy i
vi x khun v nm si.
- Dch nui cy ca cc VSV bin sau khi loi sinh khi c em th kh
nng i khng vi cc VSV kim nh bng phng php ging thch (mi
ging thch nh 200 l dch nui cy VSV).

541

- cc a thch trong t lnh 4h, sau vo t 370C trong 24h v ly ra c

kt qu. Cc chng VSV bin c ng knh vng i khng > 15mm s c s
dng tip cho cc nghin cu tip theo.
- Quy trnh sng lc s b cc chng VSV bin c kh nng sinh cc cht khng
khun, khng nm c trnh by hnh VIII.2.3.b.
VSV bin hot ha

cy im
a thch chaVSV kim nh
370C, 24h
Tch chng to vng khng

Hot trn ng thch

cy
Mi trng dch th
300 C

Vi khun 1ngy, NS v XK 7 ngy, NM 2 ngy

Dch huyn ph VSV

Ly tm

Dch nui cy VSV

Sinh khi VSV

Nh 200l

T lnh trong 4h

Ging thch, trn a cha

VSV kim nh

T m 370C, 24h

Cc chng VSV c vng i

khng >15mm

Hnh VIII.2.3.a. Quy trnh sng lc, tuyn chn cc chng VSV bin c hot tnh
khng khun khng nm
542

cc chng VSV bin to vng khng

cc chng VSV to ng knh vng

khng >15mm

Hnh VIII.2.3.b. Tuyn chn cc chng VSV bin c kh nng i khng VSV
kim nh
Sau khi tuyn chn, sng lc cc chng VSV bin c kh nng i khng
VSV kim nh, tuyn chn c 3 chng x khun VN1, VN2, VN3 v 1
chng nm si T10.6 c ng knh vng i khng > 15mm, c bit chng
nm si T10.6 c ng knh vng khng n 46mm. Cc chng x khun VN1,
VN2, VN3 c gi sang Hn Quc nghin cu, cn chng nm si T10.6
c chng ti tin nghin cu a ra quy trnh nui cy v tch chit cht
khng khun.
VIII.2.4. nh hng ca mt s iu kin nui cy ln sinh trng v sinh
cht khng khun ca chng nm si T10.6
i vi qu trnh tng hp cc cht c HTSH c rt nhiu yu t nh
hng ti qu trnh ca chng nh thnh phn mi trng, pH, nhit nui cy,
nh sng v stress. la chn c cc iu kin thch hp nht cho sinh trng
v sinh tng hp cht khng khun ca chng T10.6, chng ti ln lt nghin
cu cc yu t trn.
a. nh hng ca thnh phn mi trung nui cy
Thnh phn mi trng nh hng ln n hot tnh khng khun. Cng
nh cc loi VSV khc, nm si cng cn nhng vitamin, cht khong cho sinh
trng. Nhu cu v cht sinh trng ca mt s loi nm c th thay i ty theo
iu kin nui cy v ty theo tui ging. T bng VIII.2.4.a cho thy, thnh
phn mi trng v kiu nui cy nh hng ln n s sinh trng v sinh tng
hp cht khng khun. Chng T10.6 nui trn mi trng Czapek cho sinh khi
kh v vng khng B.subtilis ln hn so vi mi trng sinh khng sinh, nhng
li khng tt bng mi trng czapek c b sung thm bt ng, bt u tng v
cao nm men. Trn mi trng Cazek ci tin ny, sinh khi tng khong 10%.
cn ng knh vng khng khun tng 25% so vi 2 mi trng cn li. T cc

543

kt qu ny, chng ti quyt nh chn mi trng Czapek ci tin nui cy v

chn kiu nui cy tnh cho tt c nhng th nghim sau.
Bng VIII.2.4.a. nh hng thnh phn mi trng v iu kin nui cy ln sinh
trng v sinh tng hp cht khng khun ca chng nm si bin T10.6
Mi trng

Nui
cy

Lc
Czapek
Tnh

Lc
Khng sinh
Tnh
Czapek + 1%
Lc
bt u tng
+1% bt ng
v 0,2% cao
Tnh
nm men

c im sinh trng sau 7 ngy

Si nm co cm thnh vin trn nh
ht go, mu trng. Dch nui cy
mu trng,nh vng.
Nm pht trin tt, ph kn mt dch
th. Mu xanh nht. Dch mu vng
chanh
Si nm bn vo nhau to thnh
nhiu vin ht mu trng. Dch c,
mu vng
b mt mu xanh thm xen trng,
mc kn b mt. Dch trong, mu
vng sm.
Nm pht trin tt, ph kn mt dch
th. Mu xanh nht. Dch mu vng
chanh
Sinh trng rt tt, mu xanh nht
n m. Dch trong, vng mu h
phch.

11,360,2

vng
khng
B.subtilis
(d-d1)
152,2

12,380,6

301,2

10,4 0,4

152,4

11,10,5

281,4

12,682,4

26,461,4

13,210,2

39 2,12

Sinh khi
kh
(g/l)

b. nh hng ca nng mui trong mi trng

Kt qu th nghim cho thy, nng mui trong mi trng khng nh
hng n sinh trng ca chng nm T10.6. Chng u sinh trng tt mi
trng c nng mui t 0% n 3%. Lp mng nm dy, mu xanh ru thm,
ph kn b mt bnh, dch nui cy c mu vng, trong tt c cc phng n th
nghim. Nhng nng mui li nh hng n kh nng sinh cht khng khun
ca chng T10.6 rt r rt. Kt qu c trnh by trn hnh VIII.2.4.a. nng
mui 1% cho vng khng khun ln nht, nng cao hn, t 2-3% hot tnh
gim t 25-50 ln. Khi nng mui l 4% th n c ch qu trnh tng hp cht
khng khun r nht, ng knh vng v khun gim ti 85%.
c. nh hng ca pH mi trng
pH ban u trong mi trng nh hng r rt ln kh nng khng
B.subtilis ca chng nm si bin T10.6. di pH 5,5-6,5 cho vng khng khun
ln nht, t 34-37mm, pH qu kim (7,5-8,0) u lm gim vng khng khun
ca chng ti 60%. Nh vy, pH ban u l 5,5-6,5 l thch hp cho sinh tng
hp cht khng khun ca chng nm si T10.6 (hnh VIII.2.4.a). pH cui l 8,0.

544

13.5
28

12.5

30

16

25

14
12

11.3

10

20 9.2

20

15
10

40

5.5 6
4
2

30
25

2.5

8.4
8.34 8.32

8.3
8.2
8.1

15

15

10 8 8

10

7.9

7.8

Nng mui (%)

d-d1(mm)

37

20

20

0
0

34
8.3 8.3

35

pH cui

38

Vng khng khun (dd1, mm)

35

35

Sinh khi kh (g/l)

Vng khng khun (dd1, mm)

40

5.5

6.5

7.5

pH ca dch nui cy ban u

d-d1(mm)

SKK(g/l)

pH cui

Hnh VIII.2.4.a. nh hng ca pH ban u v nng mui trong mi trng

ln kh nng sinh cht khng khun ca chng nm T10.6
d. ng thi sinh trng v sinh tng hp cht khng khun ca chng nm
si T10.6
40

8.02

7.8

7.5

39.2

35
Vng v khun (d-d1,mm)

8.21

8.1

34

32

30
5.5

26

25

22

20
17

15
10

10

12.9

4
13

13.12

13.21

13.2

3
2

10.8

Sinh khi (mg/l), pH dch nui cy

45

0
0

Thi gia n (ng y)

Sinh khi

ng knh vng v khun

pH mi trng5.5

Hnh VIII.2.4.b. ng thi sinh trng, sinh tng hp cht khng khun v pH
mi trng nui cy
Nghin cu ng thi sinh trng v tng hp cht khng khun ca
chng nm si T10.6 cho thy, sinh khi kh t cao nht vo ngy th 4
(13,24g/l), cn ng knh vng khng khun t cao nht vo ngy th 7 (39
2,12). Sau , kh nng khng khun ca dch nui cy gim dn. Nh vy, t
kt qu nhn c cho thy thi gian dng ln men thch hp nht l sau 7 ngy.
Kt qu c trnh by trn hnh VIII.2.4.b.
Sau khi nghin cu nh hng cc iu kin ln s sinh trng v sinh
tng hp cht khng khun ca chng nm si T10.6 chng ti a ra quy
trnh nui cy thu dch khng khun nh sau.

545

Chng nm si
T10.6 hot ha

Mi trng
Czapek + 1% NaCl

Lc 200vng/ pht
28-300C

4 ngy
Dch ging
cp 1, 2

Mi trng Czapek 1% NaCl, 1%

bt u tng, 1% bt ng
7 ngy

Nui tnh 28-300C

Dch nui cy
chng T10.6

Dch nui cy cha

cht khng khun

Sinh khi chng

nm si T10.6

Hnh VIII.2.4.c. Quy trnh nui cy thu nhn dch khng khun t
chng nm si bin T10.6
Nghin cu tch chit hp cht khng khun
i vi vic tch chit cc cht c HTH t dch nui cy hoc dch chit,
vic la chn dung mi thch hp l iu quan trng nht. Cc dung mi c s
dng phi p ng cc tiu ch sau: (1) khng nh hng n hot tnh khng
khun, (2) c c tnh bay hi nhanh iu kin nhit thp, (3) phi tch c
2 pha r rt. Trong th nghim ny, cc dung mi c s dng trong nghin cu
l: n-butanol, 2-butanol; n-butyaxetat; ethylaxetat, metanol, isopropyl alcohol,
clorofom, diclometan.
Vng v khun ca dch nui cy chng T10.6 i vi B.subtilis sau 7
ngy l 37mm. Chng ti s dng dch ny tch cht khng khun vi 6 dung
mi: n-butanol, 2-butanol; n-butyaxetat; ethylaxetat, cloroform, diclometan. Kt
qu c trnh by trn bng VIII.2.4.b. T bng VIII.2.4.b cho thy, 6 trong 8
loi dung mi s dng c kh nng tch c 2 pha r rt. Ch c metanol,
isopropyl alcohol l khng c kh nng tch pha, nn b loi b. Nhn xt th 2
l, sau khi tch chit bng 6 dung mi ni trn th cht khng khun th vn gi
c hot tnh t 30-80% so vi hot tnh ban u. Hot tnh khng B.subtilis

546

khc nhau i vi tng loi dung mi. Nhng theo tiu ch dung mi phi bay hi
nhanh, to 2 pha ring bit, khng ln dch nui cy, chng ti chn dung mi
thch hp l cloroform. Vi dung mi ny, thi gian tch chit c rt ngn v
hot tnh khng khun vn gi c kh cao. Sau khi cho bay hi clorofom bng
my c quay, chng ti thu c dng bt vng mn, mi hc. Xc nh kh nng
khng khun ca khng sinh th cho thy, dung mi clorofom cho hiu xut tch
chit khong 70%, vng khng khun cn 26mm. T cc kt qu trnh by trn,
chng ti a ra qui trnh tch chit cht khng khun t chng nm si bin
T10.6 (hnh VIII.2.4.d).
Dch nui cy chng 10.6
tch sinh khi

Dung mi Clorofom
T l 1:1
Dung dch hn hp

Xc nh hot tnh
khng khun
Nhit
phng

o trn trong
2h
Dung dch hn hp
o trn

Nhit
phng

tnh 3h
Dung dch tch
thnh 2 pha

Nhit
phng
Pha di

Tch pha bng

phu chit
Na2SO4 10g/l

Pha trn

Hn hp clorofom + cht
khng khun
500C,
0,53 at

Dich nui cy sau khi

chit cn li

C quay
600vng/pht
Loi b

Clorofom thu hi

Cht khng khun th

Hnh VIII.2.4.d. Quy trnh tch chit cht khng khun t chng nm si T10.6
M t qui trnh: Dch nui cy tnh sau 7 ngy 300C c lc b h si
v cn mi trng. Thu c dch vng sm, trong vt. B sung dung mi
clorofom vo dch nui cy theo t l 1:1. Lc u hn hp trong 2 gi trong phu
chit. Sau hn hp dch tnh 3 ting. Lc ny hn hp dch chit to
thnh 2 pha r rt. Tch ly pha trong pha di. B sung Na2SO4 vi nng
10g/l loi b nt nc v dch nui cy. Thu c dch chit cha clorofom v

547

cht khng khun. Sau , dch chit c c quay 500C, 0,53at, tc

600vng/pht. Thu hi dung mi v hp cht khng khun th di dng bt
mn, mu vng ngh, mi hc.
Bng VIII.2.4.b. Kh nng khng khun ca dch chit khng sinh bng cc dung
mi khc nhau
S
TT
1
2
3
4
5
6
7
8

Tn dung
mi
2-butanol
n-butylaxetat
Ethylaxetat
Metanol
Isopropylalcohol
n-butanol
Clorofom
Diclometan
(dung mi
nng)

c im tch pha ca dung mi

Tch c 2 pha r rt. Ly pha trong bn trn.
Tch 2 pha kh r rt. Thu hi pha trong bn trn.
Tch c 2 pha, nhng pha trn to nh tng ln
dch nui cy.
Khng tch pha.
Khng tch pha.
Tch c 2 pha r rt, pha trong bn trn, ly pha
trong.
Tch c 2 pha, ly pha trong mu vng chanh pha
di. Dung mi bay hi rt nhanh.
Tch c 2 pha, ly pha di. Pha di to dng nh
tng, loi dch nui cy t nh tng.

Vng
khng
(mm)
24
14
25
KX
KX
26
26
24

VIII.2.5. nh tn khoa hc mt s chng VSV bin c hot tnh sinh hc

kh cao
Kt qu nh tn mt s chng vi sinh vt bin c hot tnh sinh hc c
trnh by trn bng VIII.2.5.
hiu bit su hn v cc chng VSV bin c kh nng sinh mt s cht
c HTSH nh k trn, chng ti tin hnh nh tn khoa hc ca chng. Chng vi
khun T9.5 v chng nm si T10.6 c s b nh tn theo hnh thi, c quan
pht sinh bo t v mt s phn ng sinh ho. Ring 3 chng x khun v 3
chng nm men c nh tn theo phng php sinh hc phn t.
M t c im hnh thi, vi hc ca khun lc chng nm si T10.6
Khun lc trn mi trng Czapek mc nhanh, ng knh 4-5 cm, mu
trng, mt dng xp bng dy 3 mm vng trung tm, vng st mp khun lc c
mt dng nhung, sau 12 ngy c mu xanh lc do to thnh bo t trn. Mt tri
khun lc c mu vng chanh. Khng c git tit. Gi bo t trn nhn pht trin
t si nm kh sinh hoc him hn t si nn, di 40-100 m hoc 150-250 m x
1,5-2,0 m. Thng c 2-3 nhnh dc thn gi hoc phn ngn gi. Cc nhnh
khng to thnh vng r rt. Nhnh c kch thc 5-20x1,5-2 m. Ngn gi hoc
ngn nhnh mang 1 vng th bnh 3-8 chic, th bnh 6-8x1,5-2 m, phn nh
thon dn, hi nhn. Bo t trn hnh cu, gn cu 2,5-3 m, rp nh. Theo bn

548

m t A manual of Penicilia, chng nm si bin T10.6 c tn khoa hc l

Penicillium waksmani Zaleski (hnh VIII.2.5.a).

Hnh VIII.2.5.a. C quan pht sinh bo t (a), hnh thi khun lc chng nm si
bin (b)
Bng VIII.2.5. Kt qu nh tn 11 chng VSV bin c kh nng sinh cc cht c HTSH
K hiu
Ngun
c im khun lc,
Khng VSV K
Tn khoa hc
chng
gc
sinh trng
v sinh HTSH
theo phn loi
Chng
Trm
Xm xanh, kh, vin Khng vi khun Streptomyces
x khun tch bin trng. Sinh trung tt trn Gram(+) B.subtilis aurantiogriseus
VN1
mi trng c v khng v S.aureus
c nc bin
Chng
Trm
Trng, hi xm trng. Khng vi khun Streptomyces
x khun tch bin Mt khun lc kh, bt. Gram(+) B.subtilis malayensis
VN2
Sau 6 ngy, sinh trng v S.aureus. Vng
tt trn c 2 MT c v i khng t 22 v
34mm.
khng c nc bin.
Chng
Trm
Trng, hi xm trng. Khng vi khun (+) Streptomyces
Mt khun lc kh, bt.
B.subtilis .
cheonanensis
x khun tch
VN3
Chng
Trm
Khun lc nh (u tm) , Vng khng chng Cha xc nh
x khun tch Ph trn, trng, tc sinh nm si TG3 Ph
TG1
Tam
trng chm, to thnh Tam Giang l
Giang
khng
cc vng ng tm. Trn 18mm,
vi
mi trng dch th sinh B.subtilis
trng chm. Dch nui ng knh vng
cy c mu nu en. Sau khng l 17mm
7 ngy mc kn b mt.
Chng
Trm
Mu xm bc, b mt mn Khng B.subtilis Cha xc nh
nm si tch Ph nhung, c tia hng tm. vi ng knh
TG2
vng khng 29mm
Tam
Giang

549

Chng
Trm
nm si tch Ph
TG3
Tam
Giang
Chng
Nc
nm men bin
TS1
Trng
sa
Chng
Nc
nm men bin
NT8
Nha
Trang
Chng
nm men
mu
NT5
Chng
nm si
T10.6

Nc
bin
Nha
Trang
Trm
tch bin

Chng vi Trm
khun T tch bin
9.5

Mu xanh ru, mt bt Khng B.subtilis Cha xc nh

mc st mt thch
vi ng knh
vng khng 18mm
Khun lc nh, trn,
trng, mp c rng ca.
T bo hnh ovan, trng.
Ny chi nhiu pha.
Khun lc trn, trng,
bng. T bo hnh ovan,
trng. Sinh trung tt trn
mi trng c v khng
cha mui
Khun lc trung bnh 510mm. Mu c hi. T
bo hnh elip, ny chi
nhiu pha.
Khun lc to, mu xanh
ru thm, c vng trn
ng tm. Sinh trng tt
trn mi trng c 1%
NaCl. Bo t hnh trn,
hnh ovan, mu xanh.
Khun lc trng, b mt
kh, mp hnh rng ca.
T bo hnh que ngn,
Gram (+). Sinh bo t.

Sinh axit linolenic Pichia

(18;3) kh cao, t guilliermondii
khong 6,55-8,83%
Sinh axit linoleic Candida
(18:2) cao nht, t haemulonii
28,48-38,55%,

Phaffia
Sinh carotenoid
astaxanthin, hm rhodozyma
lng 327g/g
Khng vi khun Penicillium
Gram (+) B.subtilis waksmani
mnh. Vng i Zaleski.
khng t 46mm.

Khng vi khun Bacillus sp.

Gram (+) B.subtilis
mnh, gy cht
giun t v su xanh

Hai chng nm si TG2, TG3 v chng x khun TG1 phn lp t trm

tch ph Tam Giang c vng v khun khng nm v khng khun kh cao
nhng chng ti cha xc nh c tn khoa hc ca chng.
T bng VIII.2.5 cho thy, c 3 chng x khun mc tt trn mi trng
c v khng c nc bin. Hnh thi khun lc kh ging nhau, c mu trng n
nu trng, mt khun lc kh, bt. C 3 chng u mc tt sau 6-7 ngy nui cy.
Da trn cy chng loi pht sinh (hnh VIII.2.5.b) c th nhn thy,
chng x khun VN2 c mi quan h gn gi vi loi Streptomyces malayensis
(99,78%). Trnh t nucleotids ca on 16SrDNA ca chng x khun VN2 cho
thy, chng ny c th thuc loi Streptomyces malayensis. Tuy nhin, khi kim
tra hot tnh khng VSV kim nh ca chng ny th thy chng c hot tnh
khng vi khun gram (+) Staphylococcus aureus, nhng theo ti liu th loi
Streptomyces malayensis khng i khng vi Staphylococcus aureus. Nh vy
chng ny cn nghin cu thm v mt phn loi hc, chng c th l mt loi
mi.

550

0.1
S.yatensis
S.rimosus
S.cebimarensis
S.malaysiensis
S.sclerotialus
S.erumpens
S.kasugaensis
S.rutgersensis
S.sporoclivatus
S.antimycoticus
S.indonesiensis
VN-2

Hnh VIII.2.5.b. cy chng loi pht sinh ca chng x khun VN2

- Da trn cy chng loi pht sinh, chng x khun VN1 (hnh VIII.2.5.c) c
mi quan h gn gi vi loi Streptomyces aurantiogriseus NRRL B-5416
(99,15% nucleotit trng lp).
- Chng VN3 trn cy chng loi pht sinh (hnh VIII.2.5.d) c mi quan h gn
gi vi Streptomyces cheonanensis (99,43%), Streptomyces spinoverrucosus
(97,12%). S liu 16S rDNA cho thy loi ny c th l Streptomyces sp. m c
kh nng khng nm gy bnh.
0.1
VN-1
S.coelescensICS
S.caesius
S.lienomycini
S.lividans
S.coelescens
S.marokkonensis
S.aurantiogriseus
S.cyanocolor
S.rubrogriseus
S.violaceoruber
S.humiferus

Hnh VIII.2.5.c. Cy chng loi pht sinh ca chng x khun VN1

551

0.1
S.thermocarboxydovorans
S.parvisporogenes
S.morookaensis
S.rameus
S.tricolor
S.spinoverrucosus
VN-3
S.glomeratus
S.lomondensis
S.cheonanensis

Hnh VIII.2.5.d. Cy chng loi pht sinh ca chng x khun VN3

Chng nm men bin TS1 phn lp t nc bin o Trng Sa c
nghin cu v hnh thi, nui cy, sinh l, sinh ha, xc nh h thng
Ubiquinone, mol %(G+C) v phn tch trnh t nucleotit gen 26S rDNA vng
D1/D2. Kt qu ch ra chng TS1 c c im v hnh thi, nui cy, sinh l,
sinh ha ging vi loi Pichia guilliermondii ngoi tr mt s c im: khng
ln men Sucrose, Raffinose; khng ng ha Galactose, Melibiose, Raffinose,
Inulin, D-Ribose, Ribitol, 2-Ketogluconic axit; ng ha tinh bt tan.
Gen 26S rDNA vng D1/D2 ca chng nm men bin TS1 c 560
nucleotide. Trnh t nucleotide ny 100% tng ng vi trnh t nucleotide gen
26S rDNA vng D1/D2 ca loi Pichia guilliermondii c ng k trnh t
trong ngn hng gen quc t. Do c th kt lun chng nm men TS1 thuc
loi Pichia guilliermondii.

Hnh VIII.2.5.e. Cy chng loi pht sinh ca chng nm men bin TS1

552

Chng nm men NT8 (phn lp t nc bin Nha Trang) da theo kt qu

phn loi bng phng php sinh hc phn t, chng thuc loi Candida
haemulonii
Cn chng nm men NT5 phn lp t nc bin Nha Trang, hin ang
c s dng sn xut ch phm "nm men Phaffia lm cht mu b sung
thc n cho nui trng thy sn theo cc c im phn loi sinh l sinh ha v
da vo trnh t gen 26S rDNA vng D1/D2 v cy chng loi pht sinh cng cho
thy n c quan h gn nht vi loi P. rhodozyma.
>NT5_N
TAAGCGGAGGAAAAGAAACTAACAAGGATTCCCCTAGTAACGGCGAGTGAA
GCGGGATGAGCTCAAATTTGAAATCTGGCAGCCTCCGGTTGTCCGAGTTGTAA
ACTAGAGAAGCGTTTTCCGTGCCGGCCTGTGTACAAGTCCCTTGGAATAGGGC
GTCATAGAGGGTGAGAATCCCGTCCTTGACACAGACCACCGGTGCTATGTGAT
ACGCTCTCGACGAGTCGAGTTGTTTGGGAATGCAGCTCAAATTGGGTGGTAAA
TTCCATCTAAGGCTAAATATTGGCGAGAGACCGATAGCGAACAAGTACCGTG
AGGGAAAGATGAAAAGCACTTTGGAAAGAGAGTTAAACAGTACGTGAAATT
GTTGAAAGGGAAACGATTGAAGTCAGTCATGCGTGCTCGGACTCAGCTGGGTT
CGTCCCAGTCTATTTCCGGGTGCCGCAGGTCAGCATCAGTTTCGGGCGGTGGA
AAACGGGCGGGGGAAGGTGGCATCTCCGGATGTGTTATAGCCCCCGTTTGGAT
GCATCGCGTGGGACTGAGGAACGCAGCGCGCCCTTCACGGGGTCGGTCTCCGG
ACACGAACGCGCTTAGGATGCTGGCATAATGGCTTTAAACGACCCGTCTTGAA
ACACGGACC

Hnh VIII.2.5.f. Trnh t gen 26S rDNA vng D1/D2 ca chng nm men NT5
Chng nm si T10.6 phn lp t trm tch bin Qung Bnh thuc loi
Penicillium waksmani Zaleski.
Chng vi khun T9.5 phn lp t trm tch bin c nhiu c im thuc
loi Bacillus sp.
VIII.3. KT LUN
1. phn lp v lm sch c 1052 chng VSV bin t 81 mu nc bin,
trm tch bin v ng thc vt bin Vit Nam c su, mn v khong
cch xa b khc nhau. Trong , c 552 chng vi khun (chim 52,5% tng s
chng), 253 chng nm si (24%), 157 chng x khun (14,9%) v 90 chng
nm men (8,6%). T l cc chng VSV bin phn lp t trm tch bin c kh
nng sinh cc cht c HTSH cao hn so vi nc bin. Cn cc chng phn lp
t ng thc vt bin hu nh khng c hot tnh khng khun.
2. sng lc s b v th kh nng khng 5 chng VSV kim nh (vi khun
Gram (-): E.coli; Pseudomonas aeruginose, vi khun Gram (+): Bacillus
subtilis; Streptococcus..aureus, nm men gy bnh: Candida albicans) ca
1052 chng VSV bin. Ph khng khun ca cc chng VSV th nghim rt
hp. Khng c chng no khng vi khun Gram (-), c 8,1% s chng c hot
tnh khng vi khun gram (+) v khng nm, nhng ch c 1,2% tng s chng

553

c hot tnh cao (vng v khun t 22-39 mm). Trong 4 nhm VSV bin phn
lp c, th nhm x khun c t l s chng c kh nng khng khun, khng
nm cao nht (6,8%), tip n l nhm nm si (9,1%). Nhm vi khun v
nm men hu nh khng c kh nng khng khun, khng nm, hoc c th rt
thp.
3. th kh nng dit su hi, gy c t bo ca 142 chng VSV bin. C
5,7% s chng c kh nng dit su t (Plutella xylostella) v su xanh
(Heliothis armigera). C 5,1% s chng c kh nng gy cht cho giun nhiu
t Derodigitata Miiler .
4. xc nh kh nng sinh cc enzim ngoi bo amylaza, xenlulaza v proteaza
ca 82 chng VSV bin. Trn 50% s chng c hot tnh sinh amilaza v
xenlulaza, ch c khong 12% s chng c hot tnh proteaza. Cc chng c
hot tnh enzim ngoi bo phn ln u c phn lp t cc mu nc v trm
tch ven b.
5. xc nh hm lng axit bo khng bo ho mch di ca 6 chng nm
men bin, hm lng. C 6 chng u khng sinh DHA (22:6) v ch c chng
NT2 sinh EPA (20:5) vi hm lng rt thp 0,13% tng s axit bo. Chng
NT1, NT3 v NT8 sinh axit linoleic (18:2) cao nht, t 28,48-38,55%, cn
chng NT2, TS2 v TS1 sinh axit linolenic (18;3) kh cao, t khong 6,558,83% axit bo tng s.
6. tuyn chn c 11 chng VSV bin c hot tnh sinh hc cao: 3 chng x
khun khng vi khun Gram (+), khng nm v gy c t bo (VN1, VN2,
VN3), 3 chng nm men sinh axit bo mch di khng bo ho v sinh
astaxanthin (TS1, NT8, NT5), 1 chng vi khun Gram (+) sinh c t dit cn
trng (T9.5) v 4 chng nm si khng vi khun Gram (+) (trong T10.6 c
vng v khun rt ln >45mm).
7. a ra qui trnh nui cy v qui trnh tch chit cht khng khun t dch
nui cy chng nm si T10.6.
8. tin hnh nh tn mt s chng VSV bin c HTSH cao. C 3 chng x
khun u thuc ging Streptomyces.. Chng VN1 c ti 99,15% nucleotids
trng lp vi cc nucleotit ca loi Streptomyces aurantiogriseus NRRL B5416 ; Cn chng VN2 c mi quan h gn gi vi loi Streptomyces
malayensis (99,78%), Cn chng VN3 trn cy chng loi pht sinh c mi
quan h gn gi vi Streptomyces cheonanensis (99,43%). Chng nm men
TS1 thuc loi Pichia guilliermondii. Chng NT8 thuc loi Candida
haemulonii. Chng NT5 thuc loi Phaffia rhodozyma. Chng nm si T10.6
thuc chi Penicillium waksman Zaleski. v chng vi khun T9.5 thuc chi
Bacillus sp.

554

Mt s mu vt thu thp t bin

To bin Nha Trang

Rong bin Nha Trang

San h bin Nha Trang

C, ng vt bin

C bin

S, ngao bin

555

S lng vi sinh vt bin trong cc mu nc bin

Ca sng

Gn b (<20 m)

Xa b (>20m)

A: gn b B: Xa b

Kh nng phn gii tinh bt v xenlulo ca vi sinh vt bin

556

Kh nng i khng ca vsv bin vi mt s vsv kim nh

Kh nng khng B. subtilis v S. aureus ca x khun bin

Kh nng khng nm si ca x khun bin

B.subtilis

B.subtilis

Kh nng khng VSV kim nh ca dch nui cy chng nm s bin T10.6

557

s.aureus

s.aureus

Kh nng khng VSV kim nh ca dch nui cy chng nm s bin T10.6

Mt s hnh nh v c im khun lc ca mt s
chng vi sinh vt bin

Hnh thi khun lc ca cc chng x khun bin c hot tnh khng khun

558

D. cc NGhin cu ng dng sn xut cc sn

phm t ngun dc liu bin vit nam
Mc tiu chnh ca vic nghin cu cc hot cht sinh hc t sinh vt bin
khng g khc ngoi to cc sn phm hu ch, phc v cho i sng ca loi
ngi. Vi n lc tm kim cc ng hng mi, pht trin hp l cc ngun ti
nguyn sinh vt bin, ngy cng nhiu cc sn phm hu ch ra i. Cc sn
phm ny thng tn ti di nhiu dng khc nhau nh cc sn phm dc, m
phm, thc phm dinh dng, enzim, ha cht phc v cho cuc sng in
hnh c th k n l cc loi thuc chng vi rt hay ung th nh Ara-A, AraC, manoalide cht chun c ch enzim PLA2, enzim VentTMDNA polymerase
dng trong phng php PCR t loi vi khun vng bin nc nng, dch chit
san h lm sn phm kem dng da... Tuy vy th s sn phm thc t t bin l
mt con s v cng khim tn so vi cc sn phm c ngun gc t t lin.
pht trin cc sn phm t ngun ti nguyn sinh vt bin, cn c c
nhng bc i ng n. La chn cc hng i mi nhm a cc sn phm t
phng th nghim ra th trng s tng cng kh nng s dng ngun hot cht
t i dng v ng gp vo gi tr kinh t ton cu. Nu s dng mt cch hp
l ngun ti nguyn sinh vt bin, gi tr em li s l v cng ln, ng thi
nhng hot ng ny cng gp phn thc y s thay i v kinh t x hi
cc nc ang pht trin. Tuy nhin vic phi u t vi ngun vn ln v thi
gian di a c sn phm ra th trng hin ang l nhng ro cn cho s
pht trin ca cc sn phm t i dng. Qu trnh nghin cu v pht trin cc
sn phm ny i hi s hp tc lin ngnh bao gm cc c s nghin cu, cc
nh u t, chnh sch v s quan tm ca chnh ph v cc t chc khc.
S hp tc lin ngnh s gip tp trung ngun lc ca cc chuyn gia thuc
nhiu lnh vc khc nhau v vy s ra c nhng con ng ph hp, gim
thiu chi ph nghin cu, sn xut, cc chi ph v iu tra, sng lc, x l thng
tin nhm pht trin cc sn phm mi. S phi hp ny cng s gip tng cng
kh nng tip cn, khai thc ngun ti nguyn sinh vt bin, chia s li nhun
ng thi cng gip pht trin cc kin thc chuyn mn gia cc nh chuyn
gia. C mt vi m hnh nhm pht trin cc sn phm t i dng c bit
n rng ri. Rt nhiu s hp tc gia cc cng ty t nhn v cc c s nghin
cu v pht trin cc sn phm t nhin t sinh vt bin c thc hin trong
thi gian gn y. Theo m hnh ny, cc cng ty ln kinh doanh cc sn phm t
nhin t bin s lin h vi cc nh cung cp, cc nh trung gian, mi gii cc
nc ang pht trin thu mua cc sn phm cn thit, v h cng s lin kt
vi cc c s chuyn thc hin cc th nghim sng lc, th phn ng sinh hc
hoc tinh ch cc hot cht t sinh vt bin. T , cc quc gia ang pht trin
s c ngun vn u t cho cc nghin cu tip sau, pht trin cc ngun hot
cht mi hay u t vo khai thc hp l nhm bo tn ngun ti nguyn bin.
Mt m hnh khc cng c nhc n thng c p dng cc nc pht
trin nh Chu u v M. Theo , cc vin nghin cu s thc hin cc cng tc

559

sng lc hot tnh, phn lp, xc nh cu trc cc hot cht sinh hc, th nghim
cc hot tnh thng qua cc chng trnh h tr ti chnh t chnh ph. Khi nhng
hot cht tim nng c pht hin, chng s c ng k bn quyn v ch cc
cng ty, c s c php mi c th pht trin cc sn phm ny ra th trng.
Theo m hnh ny, cc cng ty phi thit lp c mi lin h vi quc gia s
hu ngun hot cht mua bn quyn v cc iu khon kinh t khc. Nh
m cc sn phm s c bo v v vic pht trin cc hot cht ra th trng
cng em li nhng li ch khng nh cho cc c s nghin cu v quc gia s
hu ngun hot cht sinh hc .
Mt s sn phm t sinh vt bin c mt trn th trng
Tn sn phm
Ara-A

ng dng
Thuc chng vi rt

Ara-C
Axt okadaic

Thuc chng ung th

Cht chun c ch
PLA
Manoalide
Cht chun c ch
PLA2
VentTMDNA
Enzim phn ng
polymesare
chui polymerase
Formulaid
Axt bo s dng b
sung vo thc phm
dinh dng cho tr
em
Aequorin
Ch th canxi pht
quang sinh hc
Protein hunh quang Gen ch th
xanh (GFP)
Phycoerythrin
Khng th lin hp s
dng
trong
cc
phng php ELISA
Resilience
Chit sut sinh vt
bin b sung trong
m phm

Ngun gc
Bt bin Cryptotethya crypta
Bt bin Cryptotethya crypta
Trng roi
Bt bin Luffariella variabilis
Loi vi khun vng bin nc
nng
Loi vi to bin

Loi sa pht quang Aequora

victoria
Loi sa pht quang Aequora
victoria
To

San h sng
Pseudopterogorgia
elisabethae

Vit Nam, s quan tm pht trin cc sn phm hu ch t sinh vt bin l

cha nhiu. Gn y, cng vi s quan tm c bit ca nh nc v pht trin
cng ngh bin, c ngy cng nhiu cc sn phm ra i. Qu trnh pht trin
cc sn phm ny cng thuc phm vi hai m hnh nu. Chng c th c
nghin cu, pht trin t ngun ti tr ca cc cng ty, cc sn phm di dng
lin kt ny thng ch cn thi gian ngn, kh nng thu hi vn nhanh. Hn na,
chng thng tn ti di dng cc ch phm sinh hc hay thc phm b dng,
560

Kt qu thu c t vic pht trin cc sn phm l nhng minh chng sinh

ng cho hiu qu ca hng i ny nc ta. Bn cnh , nhng chng trnh
nghin cu sng lc nhm tm kim cc hot cht qu t sinh vt bin cng
c thc hin thnh cng trong nhng nm gn y. Rt nhiu hot cht in
hnh, nhng hot cht c gi tr cao c pht hin thng qua cc chng trnh
sng lc ha hc theo nh hng hot tnh. T mt s lng khng nhiu cc
sinh vt c la chn nghin cu, c n vi chc hp cht c hot tnh sinh hc
cao c pht hin, mt vi trong s chng c nghin cu, hon thin quy
trnh v ng k bn quyn v chun b cho cc giai on tip sau.
Vi li th c thin nhin u i, Vit Nam l mt trong 25 nc c
nh gi l c mc a dng sinh hc cao nht th gii. Nm trong khu vc
bin Thi Bnh Dng, t nc ta s hu hn 1 triu km2 vng bin. Kt qu
nghin cu thng k n nay thng bo c trn 12.000 loi ng thc vt bin
Vit Nam, trong c rt nhiu loi c c t hay c hot tnh sinh hc tim
tng. y l mt tin v cng thun li cho vic nghin cu pht trin cc sn
phm c gi tr cao t ngun dc liu bin di do ca nc ta. Xut pht t cc
kt qu nghin cu c bn, chng ti trnh by trong bo co ny cc nghin cu
ng dng nhm to ra cc sn phm thc phm chc nng c gi tr cao, gp phn
phc v i sng con ngi v nng cao gi tr ca ngun dc liu bin, mt
ngun ti nguyn v gi ca t nc.

561

Chng IX. NGhin cu cng ngh sn sut thc n

chc nng b sung omega 3 phng v h tr iu tr
tim mch
IX.1. m u
IX.1.1. Tnh hnh ngi mc bnh tim mch, ri lon chuyn
ho lipoprotein cholesterol cao
T chc Y t th gii (WHO), sau khi nghin cu cc s liu v bnh tt
v t vong cc nc, xp cc bnh gy t vong thnh by nhm nh sau
[313]:
A. Cc bnh tim mch nh: bnh tim bm sinh, tim do thp, tim thiu mu cc
b, bnh c tim...
B. Cc bnh h hp ( phi, cung phi, mng phi...), k c lao phi.
C. Cc loi ung th, d b phn no trong ngi cng tnh vo y.
D. Cc tai nn, ch yu l tai nn giao thng
E. Cc bnh nhim trng v k sinh trng, nh thng hn, st rt,...
F. Cc bnh thai sn (do cha v )
G. Cc bnh khc, khng nm trong 6 nhm trn

(I)

(II)

(III)

Hnh IX.1.1. T vong cc nc ang pht trin (vng trn II), cc nc

pht trin (vng trn III) v ton th gii (vng trn I) [313].
562

Bng IX.1.1. T l t vong do tng nhm bnh nm 1988 [313].

Nhm bnh

T l ton th gii

A
B
C
D
E
F
G

23%
18%
9%
6%
14%
6%
24%

T l cc nc ang T l cc nc pht
trin
pht trin
16%
21%
6%
5%
18%
7%
27%

48%
7,5%
19%
7%
18,5%
18,5%
18,5%

Hnh IX.1.2. Th t t vong do tng nhm bnh, nm 1990

v d kin nm 2020 [313].
T bng IX.1.1 v hnh IX.1.1 minh ha, ta c th thy: Trn con s t
vong ton th gii, t vong do bnh tim mch ng hng u (23%). T vong do
nhm G (cc bnh khc) c cao hn cht t: 24%, nhng l tng s ca nhiu
bnh khc khng xp c vo 6 nhm trn. Bnh tim mch ng c coi l k
git ngi s mt, trn c cc bnh h hp v cc bnh nhim trng v k sinh
trng (18% v 14%). T chc Y t th gii tnh ton rng: bnh tim thiu mu
cc b l nguyn nhn t vong s 1 trn th gii nm 1990 cng nh tng lai
2020 (hnh IX.1.2). cc nc pht trin, tc l cc nc giu c, "ngi th nht"
ca bnh tim mch cng ni bt: 48% t vong cc nc ny l do bnh tim
mch, ngha l c 2 ngi cht th 1 do bnh tim! Thnh tch git ngi ca
bnh tim mch vt xa ung th: tt c cc bnh ung th cng li ch c 19%,
ng hng th hai. Phi chng li sng giu c tha thi cc nc pht trin
lm ngi ta cht v bnh tim nhiu hn [313].

563

 M, mt thng k vo nm 2002 - 2003 lm mi ngi phi ch : C

29 giy c thm mt ngi b bnh mch vnh v c mt pht li c thm mt
ngi cht v bnh mch vnh; tng s c 60 triu ngi trng thnh ang b
bnh mch vnh do x va ng mch; t vong do bnh tim mch chim 42%
ton b cc ca t vong hng nm; ph tn do bnh ln n 128 t USD/nm [314].
Khng phi ch ngi gi hay nam gii mi l i tng mc phi nhng
rc ri v tim mch. Trn thc t, ti M, nhng bnh tim mch l nguyn do u
bng gy ra nhng ca t vong cho nhng ph n trn 25 tui, nhiu hn s ca t
vong v cc cn bnh ung th cng li. Hip hi tim mch M c tnh rng c
n 1/3 ph n ti y mc phi nhng cn bnh lin quan n tim v c rt t
ngi bit c bnh tim mch nguy him ang rnh rp h.
Trc kia, c nhng nghin cu dn n kt lun rng: nhng ph n
di 50 tui t c nguy c mc phi bnh tim so vi nam gii cng tui, cn vi
nhng nghin cu mi vi ph n tui trn 25 th nguy c t vong v bnh
tim cao gp i so vi nam gii cng tui. Trong danh sch 6 bnh d mc v
gy nguy him nht cho ph n, bnh ng v tr s 1 chnh l tim mch.
Trong cc thp k qua, nhiu nc nh Thu in, Phn Lan, c, Hoa
K... thc hin nhiu bin php phng nga bnh mch vnh v h t
c mt s kt qu kh quan. T l cht do cc bnh tim mch gy ra gim
20- 30% so vi nhng nm 70 ca th k trc. Ni chung cc bin php ny bao
gm cc li khuyn v ch dinh dng, cai thuc l, hot ng th lc v duy
tr cn nng n nh. Trong cc khuyn co v n ung, ngi ta khuyn nng
lng do cht bo cung cp khng c vt qu 30% tng s nng lng, s
dng du thc vt, tng s dng khoai, rau, tri cy. Cc loi ng ngt khng
cung cp qu 10% tng s nng lng, cn nng lng do protein nn t t 10 15%.
Vit Nam, cng vi s bng n ca th ho, s pht trin khoa hc v
cng ngh... lm thay i li sng ca ngi dn: tnh ti t vn ng, cng
thng thn kinh - tm l v ch n ung tha calo. iu ny dn n tng
nhanh s lng ngi b tha cn, bo ph, bnh nhn mc cc bnh tim mch, h
hp, bnh nhn ung th. T l ngi mc bnh tim mch v t l cht do cc bnh
ny ang chim v tr s mt.
Trong nghin cu tn thng gii phu bnh l cc trng hp va x
ng mch u thp k 80, 85% c tn thng ng mch no v 15% c tn
thng ng mch vnh.
Nm 1989, Vin Tim mch lm mt cuc iu tra ln hn, bao gm 18
tnh trong ton quc, th thy t l tng huyt p tng ln 10,52%, ngha l gp
hn 10 ln so vi 30 nm trc. Gn y nht, cuc iu tra ca Vin Tim mch
ti khu vc H Ni nm 2000 cho thy t l tng huyt p ngi ln l16%.
Nhiu ngi tng huyt p hn, tt nhin s dn n nhiu ngi cht hn do
bnh tim mch bin chng ca tng huyt p, nh chy mu no, nhi mu c
tim, cht t ngt, suy tim... [313].

564

Bng IX.1.2. T l mc v cht ca bnh tim mch ti Vin tim mch [313].
Nm

1977

1990

1996

T l mc (%)

3,8

5,34

T l cht (%)

8,8

19,2

18,73

Mt th d na nc ta l bnh nhi mu c tim, mt trong nhng bnh

tim gy t vong nhiu nht. T nm 1954 n 1962, bnh ny rt him Vit
Nam, mi nm tt c cc bnh vin H Ni v Hi Phng ch gp 1-2 trng hp.
Sinh vin trong 6 nm rng r i thc tp cc bnh vin, hu ht khng thy bnh
ny ln no. Ngay c cc bc s sut i hnh ngh cng nhiu ngi cha gp ca
no.
T 1963 tr i, tnh hnh thay i hn, mi nm s trng hp nhi mu
c tim pht trin ngy mt nhiu hn v ch trong nm 1993, Vin tim mch hc
gp 95 ca, bnh vin Hu Ngh gp 66 ca, Bnh vin Vit Tip 16 ca. thnh ph
H Ch Minh, nm 1988 c 313 ca, nhng 4 nm sau, nm 1992 tng vt ln 639
ca.
Bng IX.1.3. S lng bnh nhn nhp vin trong thp nin 90 nhng BV chnh
Tn bnh vin
BV Hu Ngh H Ni
BV Vit Tip Hi Phng
BV Thng nht TP HCM
BV Nguyn Tri Phng TP. HCM
BV tnh ng Nai
BV Thng Nht ng Nai
BV Ch Ry TP. HCM
BV ND Gia nh
BV Thng Nht TP. HCM

Nm

S ca nhp vin

1990 - 1997
1991 - 1995
1986 - 1996
1996 - 1997
1989 - 1990
1995 - 1998
1998 - 2000
1997 - 2001
1999 - 2991

275
68
149
206
24
109
63
200
82

cc nc ang pht trin khc cng c hin tng t vong do bnh tim
mch tng ln nh vy. Ngi ta c cm gic rng pht trin v kinh t thng i
km vi pht trin cc bnh tim mch. Cc nh thng k cn nhn xt rng:
tng nc, cc tng lp trn, giu c hn v sng theo "kiu phng Ty" hn,
mc nhi mu c tim nhiu hn, cc tng lp nhng ngi dn cc nc ngho
nh c cc nc giu, b nhi mu c tim nhiu hn ng bo ca h trong
nc.
Nhiu cuc hi tho v bnh l tim mch va qua ti thnh ph H Ch
Minh bo ng: s ngi tr tui mc bnh lin quan n tim mch ngy cng
gia tng. Ch trong vng mt tun l cui thng 3-2005, Vin tim thnh ph H
Ch Minh tip nhn cp cu v iu tr nhiu bnh nhn tr tui b nhi mu
c tim (36 - 42 tui). Trong t rt m ko di bt thng u nm 2008, bnh
tim mch ngi ln tui tng t bin. Trong vng mt tun, ti Bnh vin

565

Nng, bnh qun mi ngy c n 15 - 17 bnh nhn nhp vin, khoa Ni tim
mch c nhng ca rt nng. S lng bnh nhn nhp vin trong nhng ngy ny
ln n 120 ngi.
Bnh tng huyt p ( 140/90 mmHg) cng c xu hng tng nhanh, c
bit l cc i tng trn 16 tui. Theo iu tra dch t hc nm 1999 Khnh
Ho, tng huyt p t 15 tui tr ln c t l 8%, v trn ton quc nm 1993 l
11% trong nhm tui t 15 - 31 c t l mc bnh 4,3% v tng dn theo tui,
nhm tui 70 - 79 c t l mc 47,4%. Mt nhm nghin cu vn dng hng
dn ca WHO/ISH-1999 vo thc t lm sng THA ngi ln trn 60 tui ti
Khnh Ho, a ra kt lun: tng huyt p tm thu n c chim mt t l
khng nh ngi ln tui - 36,04%; tui l yu t nguy c kt hp cao nht
chim 89%, tng cholesterol ton phn > 2,5g/l (10,89%), ht thuc l 29/45 ch
yu nam gii (52%), tin s gia nh c bnh tim mch sm 5,81% [313].
Nhn chung ton cu th bnh tim mch lm ngi ta cht nhiu nht.
mt s nc pht trin, s t vong tim mch trc kia rt cao, gn y ang gim
mt cch r rt. Tri li, cc nc ang pht trin, t vong tim mch hin nay
khng ng hng u, nhng li ngy cng c xu hng gia tng. l nhng
nhn xt lm chng ta phi suy ngh.
S tng ca bnh tim mch v t l cht do bnh tim mch ngy cng cao
khin cho mi ngi bng hong v quan tm n ch n phng bnh tim
mch hn. Nhiu loi thc phm b sung omega 3, DHA v cc cht dinh dng
c tc dng ngn nga bnh tim mch xut hin v ngy cng nhiu.
Coromega l sn phm b sung omega 3 c ngun gc t nhin t du c, hm
lng EPA v DHA trong sn phm ln ti 350 v 230 mg/100g tng ng, gip
ngn nga hiu qu cc bnh tim mch. EPO (Everning Prime Rose Oil) ca c,
vi chit xut t du hoa anh tho c thnh phn axit bo omega 6 ln ti 75%
gip gim cholesterol xu v tng hm lng cholesterol tt. Ngoi ra cn c
nhiu sn phm khc cng b sung omega 3 v DHA t ngun nguyn liu t
nhin vi hm lng cao, c tc dng gim cholesterol c bit l LDL cholesterol nh: Cholesterol Reducer, Cholestasys, HTC - Lipits, Vitaheart,
Lessterol Ti Vit Nam, Vin Ho hc cc Hp cht Thin nhin - Vin Khoa
hc v Cng ngh Vit Nam cng sn xut c ch phm omega 3 - L vi
hm lng omega 3 t 40%.
IX.1.2. Ch n trong phng v iu tr ri lon chuyn
ho lipoprotein mu.
T l mc bnh tim mch khc nhau cc nc cng nh trong cng mt
nc nhng khc nhau v iu kin kinh t x hi lm cho ngi ta ch n cc
nguy c mc bnh l mi trng v dinh dng.
Theo s hiu bit hin nay ba yu t nguy c c xc nh, l ht
thuc l, tng huyt p v hm lng cholesterol trong mu cao. Trong , hm
lng cholesterol l nguyn nhn chnh gy ra cc bnh v tim mch. Ri lon
chuyn ho lipoprotein mu bao gm cc ri lon thnh phn trong chuyn ho
lipoprotein:
- Tng triglyxerit huyt tng.
566

Tng cholesterol huyt tng.

Tng Low Density lipoprotein (LDL) - cholesterol.
Gim Hight Density lipoprotein (HDL) - cholesterol.
Tng t l LDL/HDL.

Bng IX.1.4. Gii hn chun v gii hn c cho l tt ca cc thnh phn

lipoprotein huyt tng [315].

Cholesterol ton phn

Gii hn chun
(mmol/l)
3,5 - 7,8

Gii hn tt
(mmol/l)
< 5,2

LDL - cholesterol
HDL - cholesterol

2,3 - 6,1
0,8 - 1,7

< 4,0
> 1,15

Triglyxerit

0,7 - 1,8

0,7 - 1,7

Thnh phn lipoprotein

iu tr h cholesterol mu, iu chnh c cc ri lon v lipoprotein

mu khng nhng c kh nng ngn chn m cn phc hi c nhng tn
thng va x thnh mch vnh.
IX.1.2.1. Lipit v lipoprotein mu.
Lipit mu bao gm cc axit bo t do, tryglyxerit, phosphorid, cholesterol
t do, cholesterol este ho. Cholesterol l thnh t ch yu ca mng t bo v
cng l yu t chnh tng hp cc ni t nh glucocorticoid, aldosteron v axit
mt. Cholesterol trong huyt thanh khi i c vn chuyn ch yu l tiu th
LDL - cholesterol.
Tryglyxerit th ng vai tr quan trng trong vn chuyn nng lng t
thc n vo trong t bo v c vn chuyn ch yu trong cc tiu th
chylomicron v Very Low Density Lipoprotein (VLDL). Chylomicron c cu
to rut t axit bo trong thc n, c hp th qua tnh mch ca vo gan v
qua ng bch mch ngc vo i tun hon chuyn nng lng t thc n v
m m. VLDL th c tng hp gan vo t bo. Phn cn li th chuyn ho
thnh LDL - cholesterol cung cp cholesterol cho nhu cu t bo. Phn LDL cholesterol tha s c tp trung v gan v lng cholesterol trong LDL cholesterols c gii phng o thi qua mt. HDL - cholesterol cng c
tng hp gan v rut non v lm nhim v vn chuyn ngc cholesterol t
cc m v gan. Chnh v vy m khi t l cholesterol ln th c nguy c tch t
cholesterol thnh mch gy va x ng mch v thiu HDL - cholesterol,
khng vn chuyn c cholesterol v gan [316].
Cc iu tra v dch t hc cng khng nh rng lng LDL - cholesterol
cng cao th t l va x ng mch cng ln. Ngc li lng HDL cng cao th
t l va x mch vnh cng thp. T l LDL - cholesterol/HDL - cholesterol ln
hn 5 th nguy c b bnh mch vnh cng nhiu. Do hu ht cholesterol huyt
thanh l LDL - cholesterol cho nn cholesterol ton phn tng cng l mt biu
hin ca nguy c b bnh mch vnh

567

Ngoi ra nhng pht hin mi cho thy trong nhiu trng hp ch ri lon
mt thnh phn n c ca chuyn ho lipoprotein cng c nguy c dn n va
x ng mch v bnh mch vnh. Mt trong nhng yu t l ri lon
lipoprotein. nhm bnh nhn ny c th k n cc trng hp tng lipit mu
kt hp vi tnh trng huyt khi tc mch. nhng ngi n du c t gp bin
lon ny. C th du c c nhiu axit bo omega 3 c kh nng gim tnh kt
dnh ca tiu cu ngn nga hnh thnh huyt khi.
Tm li, ri lon lipoprotein mu c nhiu loi. Nguy c ch yu l gy
va x ng mch v bnh mch vnh. Cn phi ch ng bng nhng ch n
khoa hc trc khi s dng cc thuc h cholesterol trong mu [315].
IX.1.2.2. Vai tr ca dinh dng trong gim ri lon chuyn ho lipoprotein.
- Vai tr ca cht bo [315].
Tiu th cht bo c lin quan ti bnh tim mch, ung th, i tho ng,
bo ph, cao huyt p, vim xng khp. C s lng v loi cht bo n vo l
quan trng. Ch n nn gim cht bo kt hp vi gim axit bo no v
cholesterol.
- Vai tr ca axit bo no.
C mi lin quan dng tnh gia axit bo no vi nng cholesterol mu
cng nh t l mc bnh mch vnh. Nhiu nghin cu lm sng cng ch ra rng
thnh phn cht bo v s lng cholesterol ca khu phn n c tc dng ti s
thay i cholesterol mu. Axit bo no c kh nng lm tng cholesterol mu hn
l khi gim axit bo cha no c nhiu ni i. Cc axit bo no phi k n l axit
myristic (C14:0), axit lauric (C12:0)v axit palmitic lm tng cholesterol ton
phn v LDL - cholesterol.
Nhng bng chng dch t hc cho thy ch n nhiu cht bo, axit bo
no v cholesterol th c lin quan ti yu t ng mu s VII v fibrinogen,
nhng yu t ng vai tr quan trng trong hnh thnh cc mu ng, v c
xem l yu t nguy c ca x va ng mch.
- Vai tr ca axit bo th trans.
Cc axit th trans c nhiu trong m, sa ng vt n c cng c vai tr
quan trng lm tng nguy c ca bnh mch vnh. Tc dng tiu cc ca cc cht
bo th trans i vi t s cholesterol /HDL cao gp hai ln so vi cht bo bo
ho.
Axit bo th trans lm tng cholesterol ton phn v LDL - cholesterol v
lm gim HDL - cholesterol dn n lm tng t s triglicerid/HDL - cholesterol
v LDL - cholesterol/HDL - cholesterol dn n lm tng nguy c mc bnh
mch vnh. Axit bo th trans lm tng lipoprotein l cht c lin quan ti tng
yu t nguy c ca va x ng mch.
- Vai tr ca axit bo cha no.
n nhiu cht bo cha no s lm gim nguy c mt s bnh tim mch.
C mi lin quan gia mc s dng cc axit bo no trong khu phn n vi t l
t vong v mch vnh. Trong thi k 1965-1985 t l t vong do bnh mch vnh
gim Hoa K, c v Ty u nhng li tng ln ng u. Sau khi nghin
cu, cc chuyn gia u i n kt lun rng ch n l yu t thay i mi
568

trng quan trng then cht cho hin tng . S thay i hng u l c s tng
s dng cc axt bo cha no c nhiu ni i (c bit l axit linoleic v
arachidonic) so vi cc axit bo no, lm t l axit bo cha no/axit bo no tng
ln. C ch chnh ca tc dng ny l cc axit bo cha no c nhiu ni i lm
gim tng s cholesterol v LDL - cholesterol, nhng khng c tc dng r rng
vi HDL - cholesterol.
HDL - cholesterol l yu t d bo quan trng ca bnh mch vnh hn c
tng s cholesterol. Nhng ngi c HDL - cholesterol thp c nguy c cao hn
v bnh ny. Ch s d bo tt nht l t s cholesterol ton phn/HDL cholesterol . Cht bo cha no trong ch n c tc dng lm tng HDL cholesterol. Nu cc cht bo no c thay th bng Gluxit hoc cc cht bo
cha no u lm gim tng s cholesterol. Cc axit bo cha no c mt ni i
tt hn cc axit bo no nhng km tc dng hn cc axit bo cha no c nhiu
ni i i vi h tim mch.
+ Axit bo cha no mt ni i
Axit bo cha no mt ni i c th s dng linh hot trong khi xy dng
ch n v chng c th dng thay th axit bo no, Gluxit hoc cung cp nng
lng thay cho c hai. Khi ch n c nhiu axit bo cha no c mt ni i s
dn n gim cholesterol ton phn, LDL - cholesterol, triglyxerit v hn ch
n mc thp nht s gim HDL - cholesterol.
+ Axit bo cha no c nhiu ni i
Axit bo Omega - 6
Nhng th nghim lm sng nh gi hiu qu ca ch n t axit bo no,
nhiu axit bo cha no c nhiu ni i (7% nng lng) ti t l mc x va
ng mch cho thy hiu qu gim cholesterol mu ca axit bo khng no
nhiu ni i. Nhng nghin cu ny cho thy gim c ngha hm lng
cholesterol mu ca ch n giu axit bo cha no c nhiu ni i (gim 17,6
- 20% cholesterol so vi ban u). iu quan trng l gim cholesterol lin quan
n t l mc bnh va x ng mch (gim 16 - 34%).
Mt s nghin cu khuyn ngh nu mt khu phn n c 30% nng
lng khu phn t cht bo th axit bo cha no nhiu ni i nn < 10% nng
lng khu phn; mt s khc li cho rng khng nn qu 7% tng nng lng
khu phn.
Axit bo Omega - 3
Trong nhng nm gn y c nhiu cng trnh nghin cu v chng
minh v vai tr tch cc ca cc loi axit bo omega - 3 i vi phng chng cc
bnh tim mch. Cc loi c, du c cha nhiu axit bo ny nh cc
Eicosapentaenoic (EPA) v Docosahexaenoic (DHA). Cc loi thc vt ni bin
v sng h tng hp cc axit bo ny v chng l ngun thc n cho c, hi sn
v ng vt c v bin. Trong nhng nm 50 ngi ta bit vai tr gim
cholesterol ca axit bo cha no n-6-linoleic c nhiu trong thc vt v by gi
ngi ta cng nhn thy du c cng c tc dng gim cholesterol tng t. Cc
quan st trn thc a cho thy cc b tc Eskimo t l mc mch vnh thp
hn so vi ngi an Mch nh ch n ca h nhiu du c bin hn. Cc

569

nghin cu cho thy cc axit bo omega - 3 khng nhng gim cholesterol m

cn gim c triglyxerit nhng ngi c triglyxerit cao. Cc axit bo omega - 3
cn c tc dng tt phng chng lon nhp tim, rung tm tht, huyt khi v
iu chnh phn no huyt p trong tng huyt p th nh. Cc axit bo omega-3
ngun gc thc vt (axit - linolenic - ALA) cng c tc dng tt i vi bnh
tim mch. ch n giu ALA, nguy c t vong do bnh tim mch vnh gim
ti 50%. T cc quan st trn ngi ta cho rng ch n hng ngy cn tng cc
axit bo n - 3 phng cc bnh tim mch c th l mi tun nn c 2-3 ln n
c, thay th c cho tht. Tt c cc loi c v hi sn u cha cc axit bo n - 3,
ngay c khi lng lipit thp trong mt s hi sn. i vi nhng ngi khng
thch n c v hi sn c th s dng du c mi ngy 2 - 3 g.
B sung du c c tc dng lm gim c ngha triglyxerit mu trn c
nhng i tng c triglyxerit bnh thng v triglyxerit cao ( 2 mmol/l). Vic
b sung khong 9 - 13 g du c/ngy (tng ng vi 1,7 - 7 g axit bo Omega 3/ngy) th s lm gim 20 - 25% triglyxerit ngi c lng triglyxerit bnh
thng v gim 26 - 33% triglyxerit ngi c triglyxerit tng.
Ngoi ra, cc nghin cu v ch phm Omega3-L c thnh phn chnh l
cc axit bo khng no omega-3 trn ng vt thc nghim trong khun kh ti
KC09.15 cho thy: Khi dng Omega 3-L 30 ngy khng gy nh hng g n s
pht trin bnh thng v th trng v trng lng cc tng gan, lch, thn ca
chut nht trng thc nghim. Nghin cu nh hng ca Omega 3-L trn s tiu
th thc n (TA), nc ung v cht thi ca chut nht trng cho thy: S tiu
th thc n, nc ung gia cc nhm chut th nghim khng c s khc bit,
khi so snh cng thi im v lng cht thi trung bnh gia cc nhm nghin
cu khng c s khc bit, khi so snh cng thi im. Nghin cu nh hng
ca Omega3-L trn chc phn to mu v chc nng gan, thn cho thy: vi mc
liu Omega3-L dng (0,25 g/kg v 0,30 g/kg th trng) ung 30 ngy, trn
ng vt thc nghim khng thy tn thng gan, thn, lch. Hnh thi gan, thn,
lch ca nhm chng so vi nhm thuc khng c s khc bit g ng k. Thuc
khng c vi gan, lch, thn v gc m bnh hc. Hn na, ch phm
Omega3-L c tc dng hn ch s suy gim nng lc tm thn kinh trn chut
nht trng thc nghim.
Nh vy b sung du c l mt gii php an ton, c tc dng iu tr tt
cho nhng ngi b triglyxerit mu cao v gp phn ci thin tr nh.
- Vai tr ca Gluxit.
Thay th axit bo no bng lng nng lng t axit bo cha no mt ni
i hoc Gluxit u c tc dng tt vi nguy c mc bnh va x ng mch. C
cu khu phn nn c trn 55% nng lng t Gluxit. Th loi Gluxit l quan v
nn dng cc Gluxit phc hp [314].
- Vai tr ca cht x.
Ngy cng c nhiu cc nghin cu trin vng ni ln vai tr ca cht x
trong ch n vi nguy c ca bnh tim mch. Hi dinh dng lm sng ca
Hoa K khuyn co mi ngi hy tiu th s lng cht x t a dng cc
thc n ngun gc thc vt. Nhng khuyn co chung nn l n 20 - 35 g cht

570

x/ngy hoc 10 - 33 g cht x/100 Kcal. Hin ti cc qun dn c tiu th ch

t mt na nhu cu nu trn. Mt s cht x lin quan ti gim cholesterol mu.
Cc cht nhy polysaccharid hot ng trong ng rut lm gim cholesterol
mu bng cch gim hp thu cholesterol hoc axit bo hoc gim hp thu mui
mt v axit mt. Da trn 40 th nghim lm sng, nghin cu trn ng vt,
FDA ca M khuyn co: S dng cm ht ng cc l mt phn lm gim
nguy c ca bnh mch vnh tim [315].
- Vai tr ca protein
Nhng nghin cu dch t hc gn y cho thy ch n cao protin
(24% ca nng lng khu phn) bao gm protein ng vt v thc vt c tc
dng gim c ngha nguy c ca bnh tim mch. Ch l s dng tht t bo v
sn phm sa thay th cht bo [315].
- Cc cht chng xy ho.
Cho n nay hi ngh quc t v chng xy ho phng va x ng
mch thng nht rng cc cht chng xy ho c tc dng nht gm: Vitamin
E, -caroten, vitamin C, selen. Cc nghin cu thc nghim chng minh tc
dng ca cc cht chng oxy ho n qu trnh va x ng mch v nhiu cng
trnh dch t hc quan st cho thy ch n c nhiu cht chng xy ho c th
gim ti 20 - 40% nguy c bnh mch vnh [315].
IX.1.2.3. Nguyn tc xy dng ch n trong ri lon chuyn ho
lipoprotein [315].
Ch n ng vai tr trung tm trong iu tr h LDL - cholesterol mu
ngn nga va x ng mch v bnh mch vnh. Di y l mt v d v c
cu khu phn thp cht bo no v cholesterol cho bnh nhn c ri lon chuyn
ho lipit trong mu.
Bng IX.1.5. C cu khu phn thp cht bo no v cholesterol cho bnh nhn
c ri lon chuyn ho lipit trong mu.
Thnh phn ch n

Hm lng
70
10 - 15
15
5
5
5
< 100
35
Hiu qu thay i (%)
Gim 10 - 20%
Gim 10 - 20%
Tng nu c km theo gim cn
Gim nu c km theo gim cn

% nng lng t Gluxit

% nng lng t protein
% nng lng t cht bo
% axit bo no
% axit bo cha no c mt ni i
% axit bo cha no c nhiu ni i
Cholesterol (mg)
Cht x (g)
Lipit mu v lipoprotein
Cholesterol ton phn
LDL - cholesterol
HDL - cholesterol
Triglyxerit

571

IX.1.3. Mt s Nguyn liu c li cho phng nga bnh tim

mch.
IX.1.3.1. Go lt.
Mt ht thc nguyn vn c nhiu lp v, khi lp v ngoi cng (v tru)
b loi b th c c go lt. Qu trnh ny khng nh hng n gi tr dinh
dng ca ht go m cn trnh c s mt cc thnh phn dinh dng din ra
trong nhng qu trnh tip theo. Nu go lt tip tc c xt loi b lp phn
cm v gn ht lp phi th thu c go trng, nhng go ny mt i nhiu
thnh phn dinh dng. Qu trnh nh bng loi b lp alron cha cc cht bo
cn thit cho sc kho.
Go lt c gi tr dinh dng v sinh hc cao do c nhiu vitamin, cht x
tiu ho v cc nguyn t vi lng c li cho sc kho con ngi. Go lt l
ngun cung cp rt tt cc khong cht quan trng l mangan, slen v magi,
cc cht ny rt tt cho sc kho ca h thn kinh, gim hm lng cholesterol,
chng c c th s dng sn xut hoocmn.
Nhng tc dng ca go lt i vi sc kho c chng minh gim
hm lng cholesterol trong mu, tc dng tch cc ti h tim mch, hn ch cc
nguy c mc cc bnh trao i cht.
a. Gim thiu nguy c mc bnh nhi mu c tim
Thnh phn du trong go lt c tc dng gim thiu lng cholesterol.
Ngoi ra, go lt cn giu cc vitamin B. c bit ph n tui mn kinh nn s
dng go lt trong ch n ca mnh. Thng xuyn n thc phm nguyn ht,
nh go lt, t nht 6 ln trong mt tun l mt cch c bit tt cho ph n tui
mn kinh c du hiu mc cc bnh v m trong mu cao, huyt p cao, hoc cc
du hiu v bnh tim mch khc [316]. Mt nghin cu ko di 3 nm vi hn
200 ph n mc bnh tim mch c cng b trn tp ch American Heart
Journal cho thy vic n ng cc nguyn ht t nht 6 ln/tun lm chm li
qu trnh x va ng mch, lm gim tin trnh gy tc ng mch. Oxt nitric
trong go lt c tc dng tng s gin n ca mch mu v ngn nga s hnh
thnh cc gc t do v s bm dnh ca bch cu vo thnh mch mu l nguyn
nhn gy nn x va ng mch [317].
b. Kim sot huyt p
Magi gip iu chnh trng thi thn kinh v bp tht bng cch cn bng
hot ng ca canxi. Bng cch ngn chn canxi i vo cc t bo mu, magi
gi cc mch mu c ngh ngi. Nu c th chng ta thiu magi, canxi c th
tng tc i vo t do, v cc t bo thn kinh tr nn hot ng khp c th, gi
i nhiu tn hiu v gy nn hin tng co rt qu mc. V vy gy nn hin
tng cao huyt p, s co tht c, v chng au na u, tnh trng cng thng v
s mt mi. Mt cng b mi y trn tp ch American Journal of Clinical
Nutrition thng 7/2007 cng ch ra rng go lt gip ngn nga bnh cao huyt
p. Sau mi nm nghin cu pht hin ra rng ch cn n mt ba go lt mt
ngy l c th lm gim s tng huyt p n 4% [318].

572

c. Lm gim cholesterol
Cm go, lp v ngoi cng ca go lt, c tc dng lm gim lng
cholesterol xu LDL - cholesterol. Cho n nay c ch ny vn cha c lm
sng t, nhng phytat trong go lt c xem l c tc dng h tr nhiu qu
trnh bao gm qu trnh lm gim hm lng cholesterol v triglyxerit.
Most v cng s trong nghin cu nh gi nh hng ca cm go v du
cm go n hm lng cholesterol ca nhng ngi tnh nguyn c hm lng
cholesterol khng cao lm ch ra rng du cm go c tc dng lm gim hm
lng LDL cholesterol. Trong khi ch n l cm go tch bo khng lm
gim hm lng cholesterol th ch n du cm go lm gim lng LDL cholesterol n 7% [319]. Cc nh nghin cu kt lun rng nguyn nhn lm
gim cholesterol l do trong du cm go c cha mt lng hp cht phi saponin
(unsaponifiable). Cc phi saponin trong cm go rt tt i vi nhng ngi b
tim mch. Bn cnh cht x trong go lt c tc dng chng x va ng
mch.
Cc nh khoa hc tm thy cht du c trong lp cm bc ngoi go lt
cha tocotriennol c tc dng lm sch nhng cht ho hc gy nn hin tng
ng mu v ng thi gim cholesterol.
d. Lm gim nguy c ri lon chuyn ho
Theo quan nim mi, cc loi cht bo khc nhau c nh hng khc nhau
n c th, v d: cc cht bo dng bo ho v dng trans lm tng nguy c mc
bnh tim mch trong khi cht bo dng omega 3 li lm gim nguy c mc cc
bnh tim mch. Cc nghin cu gn y nht lm sng t s tng phn ny.
Hin nay, cc loi ng cc tinh ch v cc loi thc phm c lm t chng
(bnh m trng, bnh quy, cm) khng ch l nguyn nhn lm tng trng
lng c th m cn lm tng ri lon chuyn ho (bnh tim mch), trong khi n
nhiu thc phm t ng cc nguyn ht c tc dng chng li tt c cc bnh
trn. Trong mt nghin cu gn y nht xut hin trn Diabetic Care, cc nh
nghin cu phn tch d liu trn 2.800 thnh vin thy rng s ri lon chuyn
ho gim i 38% vi ngi n mt lng ln cht x t ng cc nguyn ht.
Ngc li, nhng ngi n nhiu thc phm dng tinh ch li tng 141% ri lon
chuyn ho so vi n ch yu cht x t ng cc nguyn ht
IX.1.3.2. Vng.
Vng l thc phm giu protein (20,1%), lipit (46,4%) v gluxit (17,6%),
tng ng vi lc. Vng l ngun cung cp nhiu mangan, ng, canxi, magi,
st, pht pho, vitamin B1, km, v cht x. Cng vi nhng dng cht quan
trng ny, vng cn cha 2 cht c nht v nh: sesamin v sesamolin. C hai
cht ny u thuc v mt nhm cht x c ch c bit gi l lignan, v c
cho thy c tc dng gim cholesterol trong con ngi, chng cao huyt p v
tng ngun cung cp vitamin E ng vt. Sesamin cng c bit l c kh
nng bo v gan khi tc hi oxi ha. Lipit ca vng khng ch c ngha l lipit
thc vt m n cn rt giu cc axt bo khng no mt ni i, cc axt bo
omega 3 (0,12%) v omega 6 (7,72%). Trong thc n cho ngi tim mch,

573

thnh phn khng th thiu l cc axt bo khng no, cc axt bo omega 3. Do

vy, trong cng thc sn xut thc n chc nng c tc dng gim hm lng
cholesterol trong mu, vng trng c chn lm mt trong nhng nguyn liu
chnh [320].
a. Tc dng iu tr bnh vim khp mn tnh
ng trong ht vng c tc dng gm mt s cn au v sng ty ca
rheumatoid arthritis (dng vim khp mn tnh tng dn, gy ra vim). C hiu
qu cao nh vy l v cht khong ny c vai tr quan trng trong h enzim
chng vim v chng oxi ha. Thm na, ng ng mt vai tr quan trng trong
hot ng ca lysyl oxidase, mt enzim cn thit cho vic to ra cc lin kt cho
gia collagen v elastin- cht nn to nn cu trc, sc bn v n hi ca
mch mu, xng v khp.
b. H tr sc khe tim mch v h hp.
Cc nghin cu ng h cc tc dng magie ca vng trong:
Chng co tht kh qun ngi b bnh hen suyn.
Gim cao huyt p, mt nhn t gy au tim, t qu v au tim do tiu
ng.
Ngn nga hin tng co tht mch mu ca dy thn kinh no, hin tng
ny c th gy ra triu chng au na u.
Gip ph n iu ho nhp gic ng bnh thng trong thi k mn kinh.
c. Vng lm gim Cholesterol.
Phytosterol l hp cht tm thy trong cc loi thc vt, c cu trc ha
hc gn ging cholesterol, v vi lng va trong khu phn n, ngi ta cho
rng n c th gim mc cholesterol trong mu, lm tng cc phn ng min dch
v gim nguy c ung th. Nhng tc ng c li ca phytosterol k diu n mc
chng c chit xut t u nnh, ng cc, du thng v thm vo cc thc
phm ch bin. Trong mt nghin cu trong n phm Journal of Agricultural and
Food Chemistry thng 11/2005, cc nh nghin cu cng b lng
phytosterols c trong cc loi qu v ht thng c n M. Tng lng
phytosterol trong ht vng l cao nht (400-413 mg per 100 grams), trong qu c
ch Anh v qu hch Brazil l thp nht (113 mg/100grams and 95 mg/100
grams).
IX.1.3.3. u tng.
u tng l mt loi thc phm c gi tr dinh dng cao, c bit c
nhiu protein, lipit, vitamin v mui khong. u tng cha mt lng ng k
thiamin, riboflavin, niacin, st v canxi. Hm lng lipit trong u tng kh cao
(18,4%), trong 13% l cc axt bo no, cc axt bo khng no mt ni i
chim 30% v c ti 55 60% l axt bo cha no c nhiu ni kp, ch yu di
dng cc axt bo cn thit (axt linoleic, axt linolenic) [320].
Cc nh khoa hc cn chng minh rng, u tng v cc sn phm t u
tng c cha hp cht isoflavon, c tc dng gim nguy c mc cc bnh tim

574

mch, gim hm lng cholesterol trong mu, c bit tt cho ph n thi k

mn kinh.
Mt nhm nghin cu ca Nht Bn thc hin nghin cu trong vng
13 nm v tc dng ca u tng i vi ph n v cui cng h kt lun:
Nhng ngi ph n n u n loi thc phm ny hng ngy c th gim nguy
c mc bnh tim mch t 3 - 4 ln. Nghin cu ny c thc hin vi s tham
gia ca 40.462 ngi Nht Bn bao gm c nam v n trong tui t 40- 59.
Khi bt u tham gia nghin cu, nhng ngi ny khng mc bnh ung th cng
nh cc bnh tim mch. H c chia lm 5 nhm tu theo mc s dng cc
sn phm lm t u tng mi ngy. Kt qu ca nghin cu ch r: nguy c
mc bnh nhi mu c tim hoc cc bnh tim mch l 39% i vi nhng ngi
n nhiu u tng mi ngy, trong khi nhng ngi n t u tng t l ny ln
hn 2 ln.
Da trn nhng kt qu nghin cu trong nm 1999, c quan FDA ca M
a ra khuyn co: gim cc nguy c ca bnh tim mch lin quan nn tiu
th t nht 24 g u tng/ngy. Nhng tc dng k trn l ca thnh phn
isoflavon c trong u tng. B sung 45mg isoflavon/ngy c tc dng gim c
ngha cholesterol ton phn v LDL - cholesterol nhng ph n tr tui [321].
Nghin cu trn ng vt cng cho thy isoflavon u tng c tc dng
h cholesterol ton phn v LDL - cholesterol, v tng HDL - cholesterol. Kt
qu nghin cu cho thy tiu th cc loi ch phm t u tng tng ln, c
bit l nhng loi giu isoflavon, c th dn ti h t ngt nng cholesterol
trong mu. Sn phm u tng cha cng nhiu isoflavon th tc dng n gim
nng cholesterol cng cao, iu ny khng nh rng li ch ca u tng i
vi tim l do isoflavon. Nghin cu trn 156 nam v n kho mnh c nng
cholesterol huyt ln hn hoc bng 140 mg/dl, trong 9 tun lin n u tng c
cha 62 mg isoflavon kt qu gim 4% cholesterol ton phn v 6% LDL cholesterol [321].
IX.1.4. Cng ngh p n cao p.
p n thc phm c s dng t hn 70 nm trc. Vai tr ban u
ca n rt hn ch ch phi trn v nh hnh m ng, mt loi thanh ng cc
n lin. Ngy nay, my p n thc phm c coi l l phn ng sinh hc c
nhit cao v p sut ln n chuyn ho rt nhiu thnh phn nguyn liu th
ban u thnh sn phm bin tnh trung gian hoc sn phm cui cng. ng lc
ca nhng pht trin ny ph thuc vo nhng yu cu ca cng ngh thc phm:
quy trnh c nng sut cao lin tc, hiu qu nng lng, ch bin cc nguyn
liu ban u c nht tng i cao, ci thin hng v v cu trc ca thc
phm, kim sot qu trnh trao i nhit ca cc thnh phn thc phm v s
dng cc loi nguyn liu khng truyn thng. Cc my p n 1 trc, c trang
b cc u gia nhit bng in c pht trin t nhng nm 1940 sn xut
cho n lin t bt hoc ht ng cc. Cc my p n ny c th ch bin cc
thnh phn ht ng cc nghin nh (c m ln hn 20%). Trong khi chuyn
ng qua my p n, c nng ca trc quay c chuyn thnh nhit nng, lm
tng nhit ca hn hp ln hn 1500C. Kt qu l nguyn liu b bin tnh,
575

polyme ho c y qua 1 khun. S gim p qua khun lm n sn phm v

nc c nhit cao nhanh chng b chuyn thnh hi nc.
Chc nng ca mt my p n cao p:
- Gn kt: Cc thnh phn nguyn liu c th b nn v gn vi nhau thnh tng
vin ring bit nh my p n.
- ui kh: Thnh phn nguyn liu c cha cc ti kh c th b ui kh nh
vo qu trnh p n.
- Kh nc: trong qu trnh p n thng thng mt lng m khong 4-5% c
th c loi b.
- Trng n. T trng ca sn phm ( ni hoc chm) c th c iu
khin bng cc iu kin thao tc v cu hnh ca my p n.
- H ho: Ch bin bng p n c th ci thin s h ho tinh bt.
- Nghin: Cc nguyn liu c th c nghin trong trc p trong qu trnh p
n.
- ng ho: Mt my p n c th ng ho bng cch cu trc li cc thnh
phn nguyn liu t gi tr thnh cc dng c gi tr hn.
- Phi trn: C rt nhiu loi trc p khc nhau c sn xut nhm tho mn
nhu cu phi trn vi s lng cc thnh phn mong mun trong cng mt
bung p n.
- Thanh trng v tit trng: Cc loi nguyn liu c th c thanh trng hoc
tit trng bng cng ngh p n i vi nhiu lnh vc khc nhau.
- Bin tnh protein: Cc protein thc vt hoc ng vt c th b bin tnh bng
p n.
- nh hnh: Mt my p n c th to bt k hnh dng mong mun no cho
sn phm ch bng cch thay i u khun pha cui ca khun p.
- Ct: Mt cu hnh chuyn bit bn trong khun p c th c to ra cho mt
hot ng ct mong mun ca mt sn phm c bit.
- Thay i cu trc: Cu trc vt l v ho hc c th b thay i trong h p
n.
- Lm chn bng nhit: Hiu qu lm chn mong mun c th t trong my p
n.
- ng nht: Rt nhiu dng nguyn liu c th c kt hp thnh mt sn
phm to ra cc tnh cht c bit nh s dng my p n.
Mt s li ch ca my p n trong sn xut thc phm:
- Tnh thch nghi: C th to ra rt nhiu loi sn phm khc nhau da trn s
thay i mt lng nh thnh phn v iu kin ca cng ngh p n. Qu
trnh p n c th p ng ng k s iu tit nhu cu ngi tiu dng i
vi cc phm mi.
- c tnh ca sn phm: Hng lot hnh dng, hn hp, mu sc v cc loi b
mt c th c to ra m cc phng php khc rt kh c th lm c.
- Hiu qu nng lng: Cc my p n hot ng vi nguyn liu c m
tng i thp trong khu nu chn cc sn phm, do vy, hn ch yu cu sy
li sn phm.

576

Gi thnh thp: p n l mt qu trnh c gi thnh r hn so vi cc qu trnh

gia nhit v nh hnh khc. S tit kim nguyn liu (19%), lao ng (14%)
v chi ph vn (44%) khi s dng thit b p n. Qu trnh p n cng cn t
mt bng trn 1 n v thao tc so vi cc h thng nu chn truyn thng.
Cc loi thc phm mi: p n c th lm thay i tnh cht ca protein v
tinh bt ca tht v rau v cc loi nguyn liu thc phm khc to ra rt
nhiu hoc ch 1 sn phm mi.
Nng sut cao v iu khin t ng: Mt my p n cung cp mt qu trnh
ch bin c nng sut cao lin tc v c th t ng ho hon ton.
Sn phm c cht lng cao: Do p n l mt qu trnh to ra nhit cao v
thi gian ngn, n ti thiu ho s phn hu cc cht dinh dng ca thc
phm, trong khi n ci thin kh nng tiu ho ca protein (nh bin tnh) v
tinh bt (nh h ho). p n c nhit cao cng phn hu cc loi cht phn
dinh dng nh cc cht c ch saponin, tripsin, cc loi enzim khng mong
mun nh lipaza, lipoxidaza, v vi sinh vt...
Khng c cht thi: y l u im rt quan trng i vi cng nghip thc
phm v thc n gia sc, do cc iu lut v mi trng ngy cng nghim
ngt v tn tin.
M rng sn xut: Cc s liu t qui m thc nghim c th d dng nhn ln
p dng cho h thng p n sn xut cng nghip.
S dng nh l mt l phn ng lin tc: My p n ang c s dng nh
mt l phn ng lin tc rt nhiu nc trn th gii kh hot tnh
aflatoxin trong lc.

577

IX.2. Kt qu v tho lun

IX.2.1. Nghin cu qui trnh cng ngh sn xut thc n
chc nng phng v h tr iu tr bnh tim mch qui m
cng nghip.
IX.2.1.1. Phn tch thnh phn mt s nguyn liu chnh
Trn c s nhu cu dinh dng ca ngi b bnh tim mch v tham kho
cc ti liu, chng ti la chn mt s loi nguyn liu chnh cho sn xut
thc n chc nng phng v h tr iu tr bnh tim mch, l go, go lt, u
tng, vng. Phn tch thnh phn dinh dng ca cc loi nguyn liu trn, kt
qu c trnh by bng IX.2.1 v bng IX.2.2.
Bng IX.2.1. Thnh phn c bn ca mt s nguyn liu s dng cho sn xut
thc n chc nng phng v h tr iu tr bnh tim mch
Thnh phn
Tinh bt (%)
Protein (%)
Lipit (%)
m (%)
Cht x (%)

Go

Go lt

u tng

Vng

76,2
7,3
1,4
14,2
0,6

75,2
7,8
1,3
13,2
2,9

24,6
34,0
18,4
14,0
4,5

16,6
18,1
57,6
7,6
3,4

Bng IX.2.2. Kt qu phn tch hm lng lipit v axt bo ca vng trng ( xt

v)
TT

Thnh phn

n v tnh

Hm lng

% cht kh

57,61

Lipit

Axt Palmitic

% lipit

9,48

Axt Stearic

% lipit

6,46

Axt Oleic

% lipit

38,35

Axt Linoleic (omega 6)

% lipit

43,35

Axt Linolenic (omega 3)

% lipit

2,36

Kt qu phn tch trn bng IX.2.1 v IX.2.2 cho thy go rt giu tinh bt
(76,2%), trong khi u tng l nguyn liu c hm lng protein rt cao
(34%) v vng c hm lng cc axt bo khng no rt phong ph. Cc axt bo
quan trng nh axt oleic (axt bo khng no mt ni i MUFA) trong vng
chim n 38,35% hm lng lipit. Cc axt bo omega 6 v omega 3 cng chim
n 43,35 v 2,36 % lipit tng ng. Bn cnh , go lt l loi nguyn liu rt
giu cht x, c bit lng du trong lp v cm ca go lt c tc dng lm
gim cholesterol LDL C rt tt [319]. y l cc loi ng cc sn c Vit
Nam, khi kt hp li s to ra mt ngun thc phm cn bng v dinh dng, rt
tt cho phng v iu tr bnh tim mch.

578

IX.2.1.2. Nghin cu ti u ha cc iu kin cng ngh ca qu trnh p

n hn hp nguyn liu go, go lt, u tng, vng bng ma trn ton
hc thc nghim Doehlert.
Nh trnh by phn tng quan, cng ngh p n cao p l mt cng
ngh tin tin, hin nay ang c s dng rng ri cc nc trn th gii v
Vit Nam trong sn xut thc phm nh cc loi ng cc n lin, snack. Vi cc
nguyn liu c hm lng tinh bt cao nh go hoc ng th qu trnh p n
din ra kh d dng, nhng vi cc loi nguyn liu l u tng, vng c hm
lng protein, lipit cao my rt d b tc, sn phm thng sng hoc chy.
to cho sn phm c c hm lng cc cht dinh dng cn bng, c bit l s
dng c ngun cht x t go lt, th vic la chn cc iu kin cho qu trnh
p n l rt cn thit sn phm to ra c chn, n, ho tan tt, ph
hp vi yu cu ca mt sn phm n lin (ready to eat).
Ma trn Doehlert c thit lp trn c s ca cc bin s l m ca
nguyn liu hn hp (go t + go lt + u tng + vng = 40% + 40% + 10% +
10%) trc khi a vo p n (X1), bin i t 14 - 22% v tc cp liu cho
my p n (X2), bin i t 575 - 675 g/pht ti u ha cc hm mc tiu l
ho tan (Y1) v n ca bng (Y2).
Da vo m hnh ton hc thc nghim ny, 10 th nghim c tin
hnh, trong cc th nghim mc 0 c lp li 4 ln nh gi mc
chnh xc ca cc th nghim v m hnh. Kt qu th nghim c trnh by
bng IX.2.3.
Bng IX.2.3. Ma trn thc nghim Doehlert v cc kt qu th nghim.
TT
1
2
3
4
5
6
7
8
9

Bin m ho
X1 (%)
1,000
-1,000
0,500
-0,500
0,500
-0,500
0,000
0,000
0,000

X2 (ln)
0,000
0,000
0,866
-0,866
-0,866
0,866
0,000
0,000
0,000

Bin thc
X1 (%)
22
14
20
16
20
16
18
18
18

X2 (ln)
625,0
625,0
668,3
581,7
581,7
668,3
625,0
625,0
625,0

Hm mc tiu
Y1 (%)
34,5
30,1
32,3
24,7
27,5
26,2
37,1
36,9
36,8

Y2 (Ln)
6,0
5,4
6,3
5,2
5,7
5,5
6,8
6,7
6,8

Kt qu bng IX.2.3 cho thy cc mc th nghim khc nhau th gi tr

thu c ca cc hm mc tiu cng rt khc nhau. ho tan ca bng thay i
t 24,7 - 37,1%, v n ca bng thay i t 5,2 - 6,8 ln. iu ny cho thy
nh hng ca cc bin ti cc hm mc tiu l rt ng k. Bn cnh , theo
cc kt qu tnh ton bng phn mm NEMRODW c trnh by bng IX.2.4
th tt c cc h s thch ng ca m hnh f u nm trong khong t 1-1,01.
Theo Roger (1983), i vi ma trn thc nghim Doehlert th h s f nh hn 2,5
579

l hon ton thch ng [322]. Bn cnh , cc h s r2 ( lch) ca cc hm mc

tiu Y1 v Y2 tng ng l 0,984 v 0,974, m khi r2 < 1 th phng trnh l ng
tin cy.
Kt qu tnh ton h s hi quy ca cc hm mc tiu bng phn mm
Nemrodw c trnh by bng IX.2.4.
Bng IX.2.4. Kt qu tnh ton h s hi quy ca cc hm mc tiu bng phn
mm Nemrodw
H s
Hm mc tiu Y1
Hm mc tiu Y2
hi
ho tan ca bng (%)
n ca bng (ln)
quy
Gi tr
H s
Mc ngha Gi tr
H s
Mc
(b)
thch ng
(b)
thch
ngha
(f)
ng (f)
< 0,01***

6,775

< 0,01***

0,417

0,0458***

1,819

0,01***

0,260

< 0,211***

b11

-4,650

1,01

< 0,01***

-1,075

1,01

< 0,01***

b22

-10,817

1,01

< 0,01***

-1,108

1,01

< 0,01***

b12

1,905

1,00

0,0672***

0,173

1,00

< 0,56***

b0

36,950

b1

2,950

b2

< 0,01***

Ch thch: *** mc ngha nh hn 0,01%

Phn tch phng trnh hi quy ca cc hm mc tiu

T bng IX.2.4 c c phng trnh hi quy ca cc hm mc tiu l:
Y1 - ho tan (%) = 36,950 + 2,950X1 + 1,819X2 - 4,650X12 - 10,817X22 + 1,905
Y2 - n (ln) = 6,775 + 0,417X1 + 0,260X2 - 1,075X12 - 1,108X22 + 0,173
C th nhn thy rng c m ca nguyn liu v tc cp liu cho
my p n n l u nh hng dng tnh n ho tan v n ca bng p
n:
Y1 - (b1 = 2,960 v b2 = 1,819) v Y2 (b1 = 0,417 v b1= 0,260)
Tuy nhin, nh hng ny s gim dn khi chng c gi tr rt thp hoc rt cao
trong min c kho st.
Y1 - (b11 = -4,650 v b12 = -10,817) v Y2 (b11 = -1,075 v b12 = -1,108).
Phn tch b mt p ng ca cc hm mc tiu
B mt p ng l th ca hm mc tiu c v trn h ta (XOY),
trn gi tr ca mi trc tng ng vi mt bin cn kho st.

580

Hnh IX.2.1. B mt p ng ca hm mc tiu Y1 - ho tan ca bng (%)

T b mt p ng ca hm mc tiu Y1 (hnh IX.2.1), c th thy c
min ti u cho ho tan ca bng t 37,1% l: m ca nguyn liu bin
i t 18,0 n 20,3% v tc cp liu t 620 n 640 g/pht.

Hnh IX.2.2. B mt p ng ca hm mc tiu Y2 - n ca bng (ln)

581

T b mt p ng ca hm mc tiu Y2 (hnh IX.2.2), c th thy rng

min ti u cho n ca bng t 6,8 ln khi m ca nguyn liu bin i t
18,0 n 19,4% v tc cp liu t 623 n 640 g/pht.
Kt hp b mt p ng ca hm mc tiu Y1 v Y2, c th kt lun rng
khi m ca hn hp nguyn liu ban u a vo p n t 18,0 - 19,4% v
tc cp liu t 623 - 640 g/p (tng ng 37,38 - 38,40 kg/h) th bng c
ho tan v n ln nht, tng ng l 37,1% v 6,8 ln.
IX.2.1.3. Nghin cu cng thc phi trn to sn phm thc phm chc
nng phng v h tr iu tr bnh tim mch.
Sau khi p n, hn hp go + go lt + u tng + vng c nghin
mn v phi trn vi cc loi bt tht, malto - dextrin v sau b sung ch phm
omega 3 t c bin to ra thc phm chc nng phng nga v h tr iu tr
bnh tim mch. Mc tiu ca qu trnh phi trn l to ra sn phm c t l cc
thnh phn protein, lipit, gluxit ph hp vi nhu cu dinh dng ca bnh nhn
tim mch, c bit sn phm phi giu hm lng axt bo khng no omega 3
ci thin chc nng chuyn ho lipit mu, huyt p bnh nhn tim mch. ng
thi sn phm phi c gi tr cm quan tt. Sau khi lm cc th nghim s b, mt
s cng thc phi trn c tin hnh v trnh by bng IX.2.5. Thnh phn
dinh dng ca cc cng thc phi trn c trnh by bng IX.2.6.
Bng IX.2.5. Mt s cng thc phi trn to sn phm thc phm chc nng
phng v h tr iu tr bnh tim mch
Nguyn liu

Hn hp Go + go lt +
u tng + vng (%)
Bt tht (%)
Malto dextrin (%)
Ch phm omega 3 (%)
Ph gia (%)

Cng thc
1

85

80

80

75

5
5
2
3

10
5
2
3

5
10
2
3

10
10
2
3

Bng IX.2.6. Thnh phn ho l ca mt s cng thc phi trn to sn phm

thc phm chc nng phng v h tr iu tr bnh tim mch
n v
Cng thc
Thnh phn
tnh
1
2
3
4
Tnh cht l hc
nht 500C
ho tan
Thnh phn ho hc
Protein
Lipit
Gluxit
m
Omega 3
im cm quan

Cp
%

386
36,5

391
37,1

463
39,3

558
44,5

%
%
%
%
mg/100 g
im

12,78
4,02
71,28
4,6
680,58
16,59

15,21
4,16
69,04
4,7
675,13
17,20

12,40
3,91
72,60
4,9
675,13
17,93

14,48
3,87
70,12
5,0
670,24
18,56

582

Kt qu bng IX.2.6 cho thy xt v thnh phn ho hc th c 4 cng

thc u p ng c nhu cu dinh dng cho ngi bnh tim mch [315].
l: nng lng t gluxit > 70%, nng lng t protein: 14-15%, nng lng t
lipit <15%. Tuy nhin, v tnh cht l hc, cng thc 4 c nht thp nht (558
Cp) v ho tan cao nht (44,5%). y l nhng ch s rt tt nh gi cht
lng mt sn phm ng cc n lin do nht thp v ho tan cao lm tng
kh nng hp thu ca sn phm, mt khc lm gim cm gic chng ngn khi n.
Hn na, im cm quan ca cng thc 4 cng t c cao nht (18,56 im).
Do vy, cng thc 4 c la chn cho sn xut thc phm chc nng phng
nga v h tr iu tr bnh tim mch. Tuy nhin, vi t l phi trn ch phm
omega 3 l 2%, sn phm c nh gi l cha ni bt c hng v thm
ngon do b nh hng bi mi tanh ca ch phm omega 3. Do vy, t l ch
phm omega 3 c iu chnh xung thp hn sao cho va m bo c cht
lng ca sn phm, va tng c gi tr cm quan. Kt qu c trnh by
bng IX.2.7.
Bng IX.2.7. nh hng ca t l phi trn omega 3 n cht lng sn phm
thc phm chc nng phng nga v h tr iu tr bnh tim mch
Mu

T l phi trn
omega 3 (%)

Hm lng omega 3
trong sn phm

M1

1,0

345,54

M2

1,5

457,17

M3

2,0

670,24

Gi tr cm quan ca sn phm

Thm ngon, hng v c

trng
Thm ngon, hng v c
trng
Hi c mi tanh

Kt qu bng IX.2.7 cho thy vi t l ch phm omega 3 s dng t

1,0-1,5% th sn phm thc n chc nng cho ngi tim mch vn gi c
hng v c trng. Tuy nhin, khi tng t l ch phm omega 3 ln 2,0% th sn
phm c mi hi tanh do ch phm omega 3 c chit xut t c bin v iu
ny nh hng rt ln n gi tr cm quan ca sn phm. Do vy, chng ti
la chn t l b sung ch phm omega 3 l 1,5% sn phm va c gi tr cm
quan tt, va p ng c nhu cu v omega 3 cho c th trong 1 ngy. Theo
tiu chun ca T chc Y t Th gii th nhu cu omega 3 ca con ngi l 500
mg/ngy [323].

583

IX.2.1.4. Qui trnh cng ngh sn xut thc phm chc nng phng v h
tr iu tr bnh tim mch
u tng

Sy
600C/3h

Tch v
Vng
Go
+ go
lt

u tng
mnh

Rang chn

Phi trn

p n cao p

Nghin

Hn hp bt go
+ go lt + u
tng + vng

Phi trn
Bt tht, malto-dextrin
ng gi

Thnh phm

584

Ch phm
Omega 3

Thuyt minh qui trnh cng ngh

Chun b nguyn liu:
- u tng c tch v trc khi trn vi cc nguyn liu khc bng
my xay xt.
- Go trng: lm sch, loi b sn, tru.
- Go lt: ngm nc nhit thng trong thi gian 60 pht. Sau ,
vt ra ro v trn vi cc loi nguyn liu khc.
p n: Qu trnh p n c thc hin trn my p n mt trc
- Hn hp go trng: go lt: u tng: vng c phi trn vi t l 40%
: 40% : 10% : 10% c p n iu kin m ca hn hp nguyn liu l
18-19%, tc cp liu 38-40 kg/h.
Nghin mn
Bng go + u tng + vng c em nghin mn trn my nghin ba
thu c bt hn hp.
Phi trn v ng ti
- Cc loi bn thnh phm c phi trn bng my trn theo t l: Bt
G+T+V: bt tht: malto dextrin: ch phm omga 3: ph gia = 75% : 10% :
10% : 1,5% : 3,5%.
- Thng s ca my trn: Tc vng quay bng 120 vng /pht. Thi
gian 10 pht/m 20 kg.
- Sn phm c cn nh lng 300g/ti v ng trong mng PE/Al/PP c
tc dng chng m, chng nh sng. Sau c ng hp giy. Sn phm sau
khi c kim tra cc ch tiu cht lng chnh mi c nhp kho.
IX.2.2. Nghin cu xc nh ch tiu l ho, vi sinh v th
nghim lm sng sn phm thc n chc nng phng v
iu tr bnh tim mch ti Bnh vin.
IX.2.2.1. Xc nh thnh phn dinh dng, ch tiu ho l, vi sinh, ca sn
phm
Sau khi xc nh c cc iu kin cng ngh ca qui trnh sn xut
thc n chc nng phng v h tr iu tr bnh tim mch, vic sn xut c
tin hnh. Sn phm to ra c phn tch cc ch tiu cht lng, kt qu c
trnh by cc bng IX.2.8, IX.2.9 v IX.2.10.
Cc ch tiu cm quan:
- Trng thi:
- Mu sc:
- V:
- Hng:

dng bt mn
vng sng
ngt, m
thm c trng

585

Bng IX.2.8. Cc ch tiu cht lng ch yu, gi tr dinh dng

STT
1
2
3

Tn ch tiu

n v tnh

Hm lng

%
Cp
L
a
b

41,5
542
81,36
1,21
13,74

%
%
%
%
mg/100g
mg/100g
mg/100g
%
%

4,80
71,08
15,42
3,44
457,17
39,6
2846,61
1,85
0,15

Tnh cht l hc
ho tan
nht 500C
mu

Thnh phn ho hc
4
5
6
7
8
9
10
11
12

m
Hm lng Gluxit
Hm lng Protein
Hm lng Lipt
Hm lng omega 3 (linolenic)
Hm lng DHA
Hm lng axit linoleic
Hm lng tro tng s
Hm lng tro khng tan trong
HCl 10%

Bng IX.2.9. Cc ch tiu vi sinh vt v cc c t vi nm ca sn phm

STT
1
2
3
4
5
6
7
8
9

Tn ch tiu

n v tnh

Hm lng

kl/g
kl/g
kl/g
kl/g
kl/25g
kl/g
kl/g
kl/g
g/kg

0
0
0
0
0
0
0
2 x 102
Khng pht hin

E.coli
Coliforms
S. aureus
Cl.perfringens
Salmonella
B. cereus
TSBTNM- M
TSVSVHK
Aflatoxin

Bng IX.2.10. Hm lng kim loi nng trong sn phm

STT Tn ch tiu
2
3
4
5
6
7

Ch (Pb)
ng (Cu)
Asen (As)
Thu ngn (Hg)
Km (Zn)
Cadimi (Cd)

n v tnh

Hm lng

mg/kg
mg/kg
mg/kg
mg/kg
mg/kg
mg/kg

0,02
< 0,01
< 0,002
< 0,002
8,02
< 0,01

586

Sn phm c xy dng tiu chun c s v c B Y t cp Chng

nhn tiu chun sn phm s: 6585/2007/YT CNTC vi tn gi Thc phm
chc nng: Cefish Ci thin chc nng chuyn ho lipt mu, huyt p bnh
nhn tim mch. Sn phm c bao gi trong mng PET/Al/PE, v bn ngoi l
hp Douplex. Khi lng tnh 300 g/hp (hnh IX.2.4).

Hnh IX.2.4. Nhn hp thc phm chc nng CEFISH v Chng nhn
tiu chun sn phm ca B Y t
587

IX.2.2.2. Nghin cu th nghim lm sng ti Bnh vin

nh gi chnh xc hiu qu ca vic s dng thc n chc nng n kh
nng lm gim ri lon chuyn ho lipit mu, bnh n huyt p, chng ti th
nghim lm sng trn 24 bnh nhn tim mch s dng thc n chc nng trong
vng 2 thng v ng thi so snh vi 24 bnh nhn i chng (b bnh tim mch
nhng khng s dng thc n chc nng). Sau mi thng, kim tra cc ch s lipit
mu, cc chc nng gan, thn, huyt p, nhp tim v cc du hiu c nng ca
tng bnh nhn trong 2 nhm trn. Kt qu thu c l gi tr trung bnh ca 24
bnh nhn trong mi nhm (bng IX.2.11 n IX.2.15).
Bng IX.2.11. Chc nng chuyn ho lipit mu trc v sau khi th nghim 2
nhm bnh nhn s dng thc n chc nng v nhm khng s dng thc n chc
nng
Nhm bnh
Trc th
Sau 1
Sau 2
Ch tiu
P
nhn
nghim
thng
thng
Nhm th
nghim (s
dng thc n
chc nng)
n = 24

Nhm i
chng (khng
s dng thc
n chc nng)
n = 24

Cholesterol
(mmol/L)
Triglyxerit
(mmol/L)
LDL C
(mmol/L)
HDL C
(mmol/L)
Cholesterol
(mmol/L)
Triglyxerit
(mmol/L)
LDL C
(mmol/L)
HDL C
(mmol/L)

5,5

5,1

5,1

< 0,001

2,4

2,9

2,3

> 0,05

3,3

2,6

2,5

< 0,001

1,2

1,5

1,6

< 0,001

5,5

5,9

5,8

< 0,001

2,2

2,4

2,6

> 0,05

3,4

3,5

3,5

> 0,05

1,2

1,2

1,2

> 0,05

Bng IX.2.12. S bnh nhn c ch s lipit mu nm ngoi tr s bnh thng

trc v sau khi th nghim 2 nhm bnh nhn s dng thc n chc nng v
nhm khng s dng thc n chc nng
Trc th
Sau 1
Nhm bnh nhn
Ch tiu
Sau 3 thng
nghim
thng

Nhm th nghim (s
dng thc n chc
nng)
n = 24

Cholesterol
(mmol/L)
Triglyxerit
(mmol/L)
LDL C
(mmol/L)
HDL C
(mmol/L)

588

14

10

14

12

11

Nhm i chng
(khng s dng thc n
chc nng)
n = 24

Cholesterol
(mmol/L)
Triglyxerit
(mmol/L)
LDL C
(mmol/L)
HDL C
(mmol/L)

13

13

14

13

14

12

14

13

Bng IX.2.13. Huyt p v tn s nhp tim trc v sau khi th nghim 2 nhm
bnh nhn s dng thc n chc nng v nhm khng s dng thc n chc nng
Nhm bnh nhn

Nhm th nghim
(s dng thc n
chc nng)
n = 24

Nhm i chng
(khng s dng
thc n chc
nng)
n = 24

Ch tiu

Trc th
nghim

Sau 1
thng

Sau 2 thng

Huyt p tm thu
(mmHg)

153

145

139

Huyt p tm trng
(mmHg)

90

85

85

Tn s nhp tim

82

79

80

Huyt p tm thu
(mmHg)

155

151

149

Huyt p tm trng
(mmHg)

110

107

101

Tn s nhp tim

79

80

80

Bng IX.2.14. Chc nng gan trc v sau khi th nghim 2 nhm bnh nhn s
dng thc n chc nng v nhm khng s dng thc n chc nng
Nhm bnh nhn
Nhm th nghim
(s dng thc n
chc nng)
n = 24
Nhm i chng
(khng s dng
thc n chc
nng)
n = 24

Ch tiu

Trc th
nghim

Sau 1
thng

Sau 2
thng

GOT (U/L)

30,2

31,7

25,5

> 0,05

GPT (U/L)

29,1

29,3

24,5

> 0,05

(GT (U/L)

32,5

26,5

23,5

> 0,05

GOT (U/L)

26,8

28,5

28,6

> 0,05

GPT (U/L)

26,0

28,2

29,6

> 0,05

(GT (U/L)

31,9

32,8

33,0

> 0,05

589

Bng IX.2.15. Chc nng thn trc v sau khi th nghim 2 nhm bnh nhn
s dng thc n chc nng v nhm khng s dng thc n chc nng
Nhm bnh nhn

Ch tiu

Nhm th nghim
(s dng thc n
chc nng)
n = 24
Nhm i chng
(khng s dng
thc n chc
nng) n = 24

Ur
(mmol/L)
Creatinin
(mmol/L)
Ur
(mmol/L)
Creatinin
(mmol/L)

Trc th
nghim

Sau 1
thng

Sau 2
thng

5,3

5,3

5,2

> 0,05

90,2

87,9

86,0

> 0,05

5,8

5,9

5,9

> 0,05

80,8

84,5

86,9

> 0,05

Nhn xt kt qu th nghim lm sng

- V chc nng chuyn ho lipit mu
Bng IX.2.11 cho thy bnh nhn nhm th nghim sau 2 thng c
cholesterol gim t 5,5 mmol/L (vt tr s bnh thng) xung cn 5,1 mmol/L
(nm trong tr s bnh thng), c ngha thng k (P<0,001). nhm i chng
th cng sau thi gian 2 thng ch s cholesterol khng c ci thin m cn
tng ln n 5,8 mmol/L (P <0,001).
Bng IX.2.11 cng cho thy: i vi HDL C (High Density Lipoprotein Cholesterol), l cholesterol c li th sau th nghim, tr s ny nhm th
nghim tng t 1,2 mmol/L ln 1,6 mmol/L, trong khi nhm i chng gi
tr ny khng i. i vi LDL- C (Low Density Lipoprotein - Cholesterol), l
cholesterol xu th sau thi gian th nghim, gi tr ny gim i t 3,3 mmol/L
n 2,5 mmol/L nhm th nghim, trong khi nhm i chng gi tr ny
khng c ci thin (vn gi l 3,4 mmol/L nh trc khi th nghim). i vi
ch s triglyxerit, c 2 nhm i chng v th nghim, sau thi gian 2 thng th
c s bin i khng ng k (P>0,05). Bng IX.2.12 cho thy:
nhm th nghim:
S bnh nhn c hm lng cholesterol trong mu cao (vt tr s bnh
thng) trc khi th nghim l 14 ngi, sau khi th nghim ch cn li
8 ngi.
S bnh nhn c hm lng HDL C tt (> 0,9 mmol/L) trc khi th
nghim l 21 ngi sau th nghim tng ln 24 ngi.
S bnh nhn c hm lng LDC C vt qu mc bnh thng trc
th nghim l 11 ngi nhng sau th nghim gim xung khng cn
ngi no.
nhm i chng: kt qu chuyn bin v chc nng chuyn ho lipit mu l
khng ng k. Sau 2 thng th nghim, s bnh nhn c ch s cholesterol xu
khng gim trong khi , s ngi c ch s cholesterol tt khng tng.
- V huyt p v nhp tim
Bng IX.2.13 cho thy:
nhm th nghim: Huyt p tm thu v huyt p tm trng c xu hng
chuyn v gi tr bnh thng, c ngha l chuyn bin theo hng c li.
590

 nhm i chng: huyt p tm thu v huyt p tm trng vn duy tr gi

tr tng ng nh trc khi th nghim c ngha l huyt p khng c ci
thin.
V nhp tim, c 2 nhm i chng v th nghim u khng c g bin i.
- V cc chc nng gan, thn
Bng IX.2.14 v IX.2.15 cho thy: c hai nhm i chng v th nghim
th cc chc nng gan, thn u khng c g thay i bt thng trong th
nghim. Ngoi ra, cc bnh nhn n thc n chc nng hon ton khng c du
hiu no b au bng, a lng, kh tiu, bun nn.
Nh vy thc n chc nng cho ngi tim mch c li cho vic ci thin
chc nng chuyn ho lipit mu, huyt p bnh nhn tim mch.
IX.2.3. Sn xut th nghim thc phm chc nng Cefish
Sn phm Thc phm chc nng: Cefish Ci thin chc nng chuyn ho
lipt mu, huyt p bnh nhn tim mch c sn xut trn dy chuyn thit
b ti Xng thc nghim Vin Cng nghip Thc phm vi khi lng 350 kg,
trong c 120 kg c mang i th nghim trn bnh nhn tim mch ti Bnh
vin a khoa tnh H Ty. Phn cn li gii thiu v phn phi ti Vin Cng
nghip Thc phm, Cng ty Dc phm Vit Hng.
IX.3. Kt lun
1. xy dng c quy trnh cng ngh sn xut thc phm chc nng phng
nga v h tr iu tr bnh tim mch. Cc thng s ca qui trnh cng ngh
c ti u v iu kin sn xut qui m cng nghip.
2. xc nh c cc ch tiu l, ho, vi sinh ca thc phm chc nng phng
nga v h tr iu tr bnh tim mch. Sn phm c thnh phn dinh dng cn
bng, giu omga 3 ph hp vi nhu cu ca ngi bnh tim mch.
3. th nghim lm sng sn phm trn 48 bnh nhn tim mch iu tr ti
Bnh vin a khoa tnh H Ty cho kt qu tt. Sn phm c chng nhn
tiu chun cht lng ti Cc V sinh an ton thc phm B Y t s:
6585/2007/YT CNTC vi tn gi Thc phm chc nng: Cefish Ci thin
chc nng chuyn ho lipt mu, huyt p bnh nhn tim mch.
4. sn xut c 350 kg sn phm Cefish trong 120 kg sn phm phc v
cho th nghim lm sng, 230 v ang c gii thiu cc knh phn phi
thc phm chc nng v bc u tip cn th trng.

591

Chng X. Nghin cu qui trnh sn xut thc phm

chc nng tng cng th lc cho vn ng vin
t Sao bin
X.1. m u
X.1.1. Vn ng th thao vi trao i cht v chuyn ho
nng lng trong c th.
Ho sinh hc l khoa hc, n lm sng t nhng qui lut quan trng nht
ca qu trnh bin i thch nghi v mt ho hc din ra trong t bo ca c th
con ngi v sinh gii ni chung. Ho sinh hc th thao cng vi ho sinh y hc
th thao nghin cu cc qu trnh bin i ho sinh trong c th vn ng vin
khi thc hin nhng lng vn ng th thao khc nhau. Cc kt qu nghin cu
chng minh rng chnh bn cht sinh ho hc ca cc qu trnh trao i cht
v nng lc chuyn ho ca cc h thng cung cp nng lng cho c th hot
ng trong qu trnh luyn tp th thao mi thc s l yu t quyt nh nng lc
hot ng th lc cng nh kh nng chu ng ca vn ng vin i vi lng
vn ng ln ca bi tp. Nhng bin i thch nghi trc ht phi l nhng bin
i thch nghi v mt sinh ho din ra bn trong c th vn ng vin sau nhng
lng vn ng hp l ca bi tp, c thc hin trong nhng chu k hun luyn
di hn mi l c s bn vng ca ca trnh luyn tp v trng thi xung sc
th thao. Trong qu trnh luyn tp th thao vn tiu hao nng lng v phc
hi th lc l rt quan trng. nh gi c tnh trng sc kho ca vn ng
vin th vn quan tm chnh trc tin l cc ch tiu sinh ho bn trong lin
quan n qu trnh trao i cht nh: testosteron, lactat dehydrogenaza, creatinin,
axit lactic, cc ch tiu v huyt hc.v..v... Ngoi ra ngi ta cn nh gi cc ch
tiu bn ngoi nh v tm l, phn x thn kinh, chuyn mn s phm... [324329]
X.1.2. Hot cht thin nhin trong thc phm l ngun
dinh dng v l thc phm Thuc c gi tr v rt quan
trng vi sc kho con ngi.
Cc hoocmn steroit, protein, axit amin v cc cht vi khong (Cu, Fe,
Zn..) c vai tr rt quan trng trong c th con ngi. Nhng cht ny u c
tng hp qua con ng chuyn ho nng lng ca c th, chng c nhng chc
nng v nhim v ring.
Vai tr v tc dng ca cc protein, axit amin trong c th c tng hp
trc tip qua ng thc phm b sung dinh dng.
Chng ta u bit rng c th ngi u nhn cc protein v cc axit amin
t thc n. Vai tr ca cc axit amin trong c th rt quan trng nh lxin l cc
axit amin gi vai tr trong tng hp protein c bp v hot ng ca tuyn tu, rt
cn cho s tun hon ca mu. Izolxin cn thit cho qu trnh sinh tng hp
glucogen v cc xetonic, nu c th thiu axit amin ny s dn n vic gim st
cn nhanh.
Lysin gi vai tr trong sinh tng hp hemoglobin, axit nucleic, nh hng
ln cc hot ng thn kinh, tiu ho v vic hnh thnh cc m xng.
592

Theonine c tc dng chng m ho gan. Prolin v axit glutamic c tc

dng tt trong vic trao i cht ca c th.
Mi mt lin kt gia cc axit amin li c nhng vai tr v chc nng
ring i vi c th. mt s peptit, isopeptit c trng lng phn t thp nh
(Lys-iso-leu, phe-his-met, gly-acg-met) v cc amino axit c tc dng tng kh
nng min dch, chng oxy ho trong vic hi phc th lc v tng cng th lc.
Cng thc cu to v trng lng phn t ca 20 axit amin:

593

Vai tr ca cc nguyn t vi khong trong c th khi c b sung qua ng

thc phm Vin tng lc.
S ng gp quan trng ca cc nguyn t vi khong l rt cn thit.
Ngi ta v nguyn t km (Zn) trong c th ngi nh l ngn la sinh mng.
St (Fe) tham gia vo qu trnh to mu. St (Fe) v km (Zn) ng vai tr quan
trng v cn thit trong cc hot ng chc nng ca gan, thn, tuyn tu, tuyn
sinh dc v cc c quan th gic. N rt cn cho s kch hot enzim
lactatdehydrogenaza phn gii cc axit lactic gii phng cc axit lactic ra khi
t bo c, chng mt mi, chng oxy ho trong c th.
Hm lng mui v c trong c th gm by nguyn t: Ca, Ka, Na, P, S,
Cl. Cc nguyn t khc: Fe, Cu, I, Zn, Mg, Se, v.v.. Cc loi trn chim mt
lng rt nh trong c th, gi chung l nguyn t vi lng.
Cht st (Fe) c thc n cung cp cho c th. Thiu st no b s khng
c cung cp oxy, khng nng lng. Nhiu nghin cu xc nh ch s
thng minh (IQ) ca tr t l thun vi hm lng st trong mu dy rn ca b
m. Khi b m thiu ht cht st, ch s pht trin ca no b gim thai nhi v
tr di mt tui. Thiu st no b s khng pht trin ng mc. ngi ln
thiu st dn n thiu mu v d mt mi.
Ngun cht st : C th con ngi hp thu km cht st. Ch 2-3% cht st
trong thc vt c c th hp th (u kh, cacao.) Cht st ng vt c
hp thu d dng hn: T 20-30% (tht , di trng, gan, tim, bu dc, tht th,
c thu, s c, c cm, c mi v cc ng vt bin).
Cc thc phm ngun gc ng vt l ngun cung cp tt nht cht st v
vitamine cn thit cho no b v s hnh thnh ca cc mng t bo, cho cc
nron, cc cht trung gian thn kinh. [328, 330, 331]
Cc nguyn t vi lng ny rt cn thit cho c th, song bn thn n li
khng sinh ra nng lng. Mui v c rt quan trng, v d: Ca l thnh phn
chnh cu thnh xng v rng, gi vai tr quan Cc loi trn chim mt lng
rt nh trong c th, gi chung l nguyn t vi lng.
Cc nguyn t vi lng ny rt cn thit cho c th, song bn thn n li
khng sinh ra nng lng. Mui v c rt quan trng, v d: Ca l thnh phn
chnh cu thnh xng v rng, gi vai tr quan trng c bp, thn kinh; P, S l
thnh phn cu thnh anbumin; Na, K, Cl duy tr thm thu protein vi t bo.
594

Mui v c c li cho sc kho hai im:

- Nu nh c th thiu mt nguyn t vi lng no , dn ti kh nng
vn ng gim, khi c b sung, c th nhanh chng hi phc nh ban u.
- Nu lng b sung thch hp, c th s pht trin tt hn. Nu b sung
mt lng thch hp s tng kh nng min dch.
Cc nguyn t in gii: Na, K, Cl. Nu tp luyn v thi u trong ma h,
m hi ra nhiu, dn ti hin tng thiu ht mui v c.
+ Nguyn t Na: Tp luyn lin tc trong mi trng nng dn ti thiu Na. Mt
Na s mt nc, biu hin: c bp co git, au dn, mt lc, v.v.. Nu gim th
trng, i i tin nhiu, tiu tin t, da ph n. Cn b sung Na.
+ Nguyn t K: Hm lng K trong mu thp, gy mt mi. Tc dng ca
nguyn t vi lng ny ngy cng c nhiu ngi ch ti, lng tuy nh song
khng nn coi thng. Se, Zn, Cu v.v.. u l cht mi gii quan trng. Khi c th
thiu Zn hoc thiu Fe, chc nng min dch gim.
Bng nhiu con ng khc nhau, mt s nguyn t vi lng gip c th
thc hin tt chc nng ni tit, tun hon mu, mau lnh vt thng, khng ch
t bo gy nhiu. Zn c chc nng duy tr sinh l ho trong c th, tng kh nng
sinh dc cho nam gii, gim bt cc nguyn t vi lng t do. Cc b phn trong
c th d b tn thng: gan, da, mt, v.v, Zn sau khi c b sung, c cng hiu
ngay.
Cng c nhng nguyn t vi lng, thng qua c ch phn ng ca cc
gc t do v tc dng qua oxy ho (peroxid) ca cc cht bo m c th hn ch
c hng lot nhng ri lon chc nng ca t bo; th d (Zn) l nguyn t vi
lng rt quan trng trong vic duy tr chc nng sinh l, sinh ho bnh thng
ca t bo v gim nh tc hi ca cc gc t do. Nhiu kt qu nghin cu
pht hin thy nhng t chc, c quan trong c th d b oxy ho gy tn thng
nh gan, da, mt, tinh trng....nng km trong cc t chc t bo trn m km
s sinh ra mc bnh, nh sinh khi u, nam gii thiu nng sinh dc v.v... sau khi
b xung thm km thng t c hiu qu iu tr rt tt. Km c coi nh l
thuc chng oxy ho (antioxydant) quan trng, iu ny d c nhiu hc gi
cng nhn.
Selen l nguyn t vi lng tham gia vo cu trc Coenzym(nhm phprosthetic, phn phi protid, l nhng cht hu c c hiu, Mangan l Cofactor
ca nhiu men gan cc men, cc men chng oxy ho glutathione peroxydaza, c
c tnh chng oxy ho v tc dng quan trng trong iu tit nng lc min dch.
Selen tham gia bo v n ron thn kinh. Magi c tc dng kch thch thn kinh,
chng trm cm
Vitamin c th chia thnh 2 loi chnh: Vitamin tan trong m v vitamin
tan trong nc. Vitamin tan trong m bao gm cc loi: vitamin A, E, D, K; cn
vitamin tan trong nc. bao gm cc loi: vitamin C, vitamin nhm B. T l bi
tit ca vitamin bo khng cao, nu thu np qu nhiu lm tch t trong c th s
c kh nng gy ra nhng nh hng xu. Cn t l bi tit ca cc vitamin tan
trong nc kh cao, ni chung khng b tch t trong c th, nu s dng vi
lng ln, thng thng rt t gy c tnh.

595

Vitamin A c quan h mt thit n th gic, chc nng sinh l th 2 ca

vitamin A gn vi s hnh thnh t bo biu b v gp phn lm lin vt thng.
Khi c th thiu vitamin A s dn n s suy gim ca kh nng thch ng trong
bng ti ca mt v chng m m, tip theo s ko theo hng lot bnh trong
qu trnh hnh thnh m biu b nh: kh da, da kh rp khc thng, hnh thnh
vy.
Tc dng ch yu ca vitamin E, C l chng s oxy ho, c kh nng lm
gim bt nhng tc hi ca oxy ho lipit. Vn mu cht trong qu trnh thay
th nng lng v vt cht trong c th l s tham gia ca nhiu loi vitamin, c
bit l cc vitamin nhm B. Khi c th thiu mt loi vitamin no s gy cn
tr cho qu trnh thay th nng lng, nh hng xu ti ton b c th, gim tc
sn sinh nng lng, cng nh th lc.
Vitamin B1 (thiamine), vitamin B2 (lactoflavine), niacin l nhng vitamin
c lin quan mt thit ti qu trnh thay th nng lng trong c th, nhu cu v
cc cht ny t l thun vi tng nhit nng c th hp th. V vy, ni chung
thng ly tng s Calo nhit lng c cung cp mi ngy lm n v biu th
lng cn thit v 3 loi vitamin.
Vitamin B6 (pyridoxil) c quan h rt cht ch vi qu trnh thay th axit
amin, do lng cn thit tng gim tu theo lng protein c hp th.
Trong c th vitamin khng hay b thiu l mt qu trnh din bin
chm chp. Khi trong thc n thiu mt loi vitamin no , biu hin u tin l
lng vitmin d tr trong m gim st, tip theo xut hin s thiu ht vitamine
v nhng hin tng sinh l bt thng, dn dn gy nn s thiu ht trn phng
din t chc (tc cu trc), v cui cng l cc biu hin lm sng. S thiu ht
vitamin nh trong thi gian di (hay gi l gn mc thiu ht) khng nht
thit xut hin cc triu chng lm sng, nhng c kh nng lm gim hiu sut
lao ng (bao gm c lao ng tr c), ng thi ko theo cm gic kh cng s
suy gim sc khng. V vy, cung cp vitamin mt cch hp l thng qua thc
n khng nhng c kh nng phng bnh m quan trng hn l n gip ta khng
ngng tng cng sc kho.
Vic b sung cc cht dinh dng c ngun hot cht t nhin cho vn
ng vin nhm mc ch gi cho th trng vn ng vin lun trng thi cn
bng v hng phn trong tp luyn
v thi u, cn bng cc cht dinh dng trong c th bao gm:
S cn bng gia 3 loi cht dinh dng sn sinh ra nhit lng. T l
nng lng c to ra nh protein/lipit/ng khong 1/1/4
S cn bng gia cc loi mui v c v nguyn t vi lng: Zn v Fe c
tnh cnh tranh mnh. Nu b sung qu nhiu Fe c th lm gim t l hp th Zn
trong c th gy nn tnh trng thiu Zn.
S cn bng gia cc loi vitamin, cn bng gia protein v axit amin: nu
cung cp qu nhiu hay qu t mt loi axit amin no s cn tr tc dung ca
mt s axit amin khc. Trong qu trnh c th tng hp protein, nhu cu v axit
amin khnng hon ton ging nhau. Lng protein ph hp vi t l axit amin
c gi l protein u cht.

596

X.1.3. Testosteron v cc hoocmn steroit vi s mt mi

trong luyn tp th thao v hun luyn qu sc v tc
dng ca hoocmn steroit trong c th c b sung qua
ng thc phm chc nng.
Testosteon, progesteron, estradiol, cortisol l cc hoocmn steroit tan
trong lipit, c th i qua mng ca t bo d dng th th th l cc protein c
hiu khu tr nm trong t bo ( bo tng v nhn t bo), nhng protein c
trng lng phn t thp nhng c c hiu cao v i lc rt ln. Cc hoocmn
steroit bn vng, c cu to ho hc thng gn ging nhau.
OH

OH

HO

Testosteron

Estradiol

COCH2OH

COCH3

OH

HO

Progesteron

Cortisol

Hnh X.1.3.a. Cng thc cu to ca cc hoocmn steroit

Cc hoocmn steroit (testosteon, progesteron, estradiol, cortisol) rt k
nc, tan d dng trong mu, c vn chuyn di dng kt hp vi protein vn
chuyn c hiu t ni chng c gii phng n c quan ch. Ti c quan
ch, nhng hoocmn qua mng bo tng bng c ch khuch tn n thun v
ri gn vi th th c hiu trong nhn (hoc th th bo tng) to thnh phc
hp hoocmn cht th th. Mt t bo ch cha khong 10.000 th th steroit.
Cc th th c gia phn t mt vng lin kt vi AND ca nhn t bo v mt
vng lin kt vi hoocmn. Phc hp phn nhy cm vi hoocmn (hoocmn
responsive element). S kt hp ca phc hp hoocmn - cht th th trn vng
AND s kch thch v khi ng ARNm. ARNm ra bo tng v cm ng s tng
hp mt hoc nhiu protein mi bo tng c hiu cho s p ng ca
hoocmn.

597

Hnh X.1.3.b. C ch tc dng ca hoocmn steroit trn t bo ch

Nhp bi tit ca hoocmn l nt chung ca h thng ni tit. S ri lon
nhp bi tit hoocmn cng l nguyn nhn chung v bnh l ni tit. Nhp bi
tit hoocmn (hay chu k bi tit hoocmn) c th thay i:
- Theo gi: bi tit LH v testosteron.
- Theo ngy: thay i ngy m i vi bi tit cortisol.
- Theo thng: Chu k kinh nguyt.
- Theo ma hoc thi gian di hn: Bi tit thyroxin..
Cortisol kch thch tng hp glucogen, l steroit chuyn ho ng, c tc
dng ln chuyn ho Gluxit, kch thch s tn to glucose t nhng axit amin, c
tc dng lm tng kh nng min dch, chng vim nhim. Cortisol v ACTH u
c bi tit vo 8 gi sng, iu ny gii thch nh sau: Trong na thi gian ngh
hng ngy (Sau 1 gi n 2 gi bui sng) xut hin hot ng ca h thng CRH
ACTH Cortisol nhm m bo cho c th c nng glucose mu bnh
thng trc khi bt u bc vo s hot ng hng ngy v trong khi cha c
ba n u tin ca ngy.
+ Chuyn ho Gluxit: Tng hp thu glucose rut, tng phn hu
glucogen.
+ Tng phn hu lipit nht l triglyxerit, phospholipit v cholesterol.

598

+ Tng tng hp protein do tc ng trc tip n s hot ho RNA

polymease hoc gin tip qua kch thch bi tit GH.
+ Tng cng s dng oxy ca c th, tng chuyn ho c bn, c tc
dng sinh nhit

Hnh X.1.3.c. C ch tc dng ca testosteron

Testosteron c mt c ch tc dng c bit v trong a s t bo ch,
testosteron tc dng nh mt tin hoocmn (pre-hoocmn). Mt enzim 5reductase ca bo tng t bo ch chuyn mt cch khng thun nghch
testosteron thnh dihydrotestosteron (DHT).
OH

OH

5-reductase

O
H

Testosterone

5-Dihydrotestosterone

Hnh 1.3.d. Chuyn ho testosteone thnh dihydrotestoster

nh enzim 5-reductase
Testosteron l hoocmn c hai tc dng: Tc dng nam tnh v tc dng
chuyn ho. Testosteron va l mt hoocmn va l mt prohoocmn. c tc
dng ln t bo ch th trc tin n c chuyn thnh dihydrotestosteron
(DHT), n c chc nng bit ho sinh hc trong qu trnh pht trin phi, m
bo s trng thnh, dn ti dy th v bo tn c tnh sinh dc cng nh bn
nng sinh dc ca n ng (ging c), m bo sinh sn ngi nh s to tinh

599

trng, c vai tr kch thch to hng cu, tng hp Hemoglobin trong nguyn
hng cu.
Luyn tp th thao lm thay i rt ln nng testosteron trong mu.
Mi quan h gia ni tit t nam tnh Testosteron v hot ng th thao c
nhiu nh nghin cu quan tm n t nhng nm 1970. Cho n nay c rt
nhiu cng b khoa hc v cc cng trnh nghin cu v vn ny [332] .
Testosteron l kch t ging c, dn xut ca steroit c cha 19 nguyn t
carbon v l mt trong nhng kch t ng ho ch yu ca c th. Ni tit t ny
c mt th bc cao trong s iu ho qu trnh trao i cht. Testosteron trong c
th tng hay gim u gy ra s thay i cc cng sut ca cc h thng trao i
cht v chuyn ho nng lng. T thc tin c cc nh khoa hc chng
minh khi c th trng thi xung sc th hm lng testosteron lc ny t
c n nh cao v ngc li khi c th trng thi mt mi th hm lng
testosteron trong mu lc ny b tiu gim [324, 329, 333-335].
* FFA : Axit bo t do
Hm lng testosteron ngi tu thuc vo gii tnh v cc la tui khc
nhau. Hm lng testosteron n gii thp hn nam gii khong 10 ln. Theo
L c Trnh [335] th hm lng testosteron trong mu ca nam v n gii cng
nh cc la tui l khc nhau. Bng X.1.3 v hnh X.1.3.e l tng kt nhng sai
khc v hm lng testosteron cc nhm tui khc nhau.
Bng X.1.3. Hm lng testosteron trong mu nam gii cc la tui khc nhau
(nmol/l).
Nhm tui

Testosteron

Nhm tui

Testosteron

4 - 10
10 - 12
12 - 14
14 - 16

0,35
1,04
3,13
7,29

16 -18
20 - 30
30 - 40
40 - 50

10,42
22,92
19,44
16,32

Hnh X.1.3.e. Nhp ngy m ca testosteron ngi tr v ngi c tui

600

Hnh X.1.3.f. Nhng tc dng chnh ca cortisol trong chuyn ho trung gian.

Hnh X.1.3.g. Nhp ngy m ca ACTH v cortisol ngi bnh thng

Theo cc kt qu iu tra nghin cu ca Vin Khoa hc Th dc Th thao
[327, 328] th hm lng testosteron trong mu vn ng vin cao hn ngi bnh
thng t 1,5 - 2,5 ln.
Khi mc testosteron trong mu gim r rt v lu khng hi phc c cn
tnh n kh nng lng hoocmn khng hoc chc nng ca trc HPG
(Hypothalamus-Hypophysis-Gonad) suy gim. vn ng vin th thao hm
lng testosteron trong mu gim di 0,69 nmol/l i vi n hay di 3,47
nmol/l i vi nam th c coi l trng thi in hnh luyn tp qu sc v lc
ny cn c tng cng b sung ti to hi phc [325-328, 336].

601

Hnh X.1.3.h. Thiu s dao ng bnh thng v sng v chiu

ca cortisol mu trong bnh Cushing
X.1.4.
Mi
lin
quan
gia
Testosteron,
Lactat
dehydrogenaza (LDH) v axit lactic trong vn mt mi
v hi phc ca vn ng vin.
Mt vn rt quan trng trong dy chuyn phn ng sinh ho trong c
th l khi hm lng testosteron trong mu tng th t l thun vi hot enzym
LDH tng km theo s gim xung ca axit lactic. Enzym LDH tham gia vo qu
trnh gii phng axit lactic trong c, m nh ta bit axit lactic chnh l nguyn
nhn gy ra mt mi ca c th. Trong th thao axit lactic l ch tiu c trng v
c tin cy cao trong vic theo di v khng ch lng vn ng thch hp ca
vn ng vin [324, 327, 328, 329, 331, 332, 337, 338]
Axit lactic l sn phm ca qu trnh ng phn ym kh (glycolysis).
Phn ng c din ra nh sau:
CH3.CO.COOH + NADH -----> CH3.CHOH.COOH + NAD
(axit pyruvic)
(axit lactic)
trng thi yn tnh, trong iu kin oxy, ti cc c quan, t chc
trong c th vn din ra qu trnh phn gii ng ym kh mnh m v sn sinh
ra axit lactic (AL) v axit ny mau chng i vo mu v vy trong trng thi yn
tnh, trong mu lun lun duy tr mt mc axit lactic nht nh. Trong lc ngh
ngi, nng AL trong mu ng mch khong 0,4-0,8 mmol/l. Gia vn ng
vin v ngi bnh thng khng c s khc nhau v lng AL lc tnh. Tuy
nhin trong qu trnh tp luyn v thi u c s khc bit nhau qu xa, bi v kh
nng chu ng ca vn ng vin i vi nng axit lactic cao trong mu l
yu t c ngha quyt nh t c thnh tch cao. Cng tc hun luyn vn
ng vin ng ngha vi vic hun luyn cho vn ng vin tp chu ng
ngng axit lactic cao trong mu cc mc thi gian di, ngn khc nhau
[325, 326, 329, 332, 334].

602

X.1.5. Thiu mu do mt mi trong luyn tp, thi u qu

sc v bin php hi phc.
Thiu mu trong th thao l dng thiu mu c tnh tng i VV ch
yu l thiu mu do thiu st (thiu mu do nhc sc) [325, 326, 335, 339].
Trong qu trnh luyn tp th thao lng st hp th gim mnh, lng st mt i
theo mc thot m hi. vn ng vin bng trung bnh lng st mt
theo m hi c th ln ti 0,6mg/l, la tui nhi ng l 0,4mg/l. V vy trong
hun luyn v thi u vn ng vin lun cn phi quan tm dn vn b sung
st nh succinate st, lactate st, ferro (Fe2+) sulphate..v..v.. nhm nng cao d tr
protein cha st. Ni n thiu mu ca vn ng vin l ni n s gim st mc
hemoglobin so vi nhu cu pht huy nng lc vn ng. Tu theo mc pht
trin ca trng thi luyn tp qu sc m c th t thiu mu ca vn ng vin,
h thp xung ti mc ch tiu thiu mu ca ngi thng v t thiu mu gy
gim st nng lc vn ng s dn n pht trin thnh thiu mu bnh l. Do
vy hemoglobin l mt ch tiu quan trng nh gi th lc ca vn ng vin
[330, 332, 333, 340].
X.1.6. ng dng cc bi thuc dn gian v c truyn vo
vic b sung dinh dng v thc phm thuc nhm tng
cng th lc cho cc vn ng vin Vit nam.
Trong iu kin bnh thng, nng lng c cung cp v b tiu hao
lun gi mc n nh. Nng lng l iu kin c bn ca mi hot ng trong
c th. Nng lng khng cc hot ng chc nng gim st, sc kho km;
nng lng qu tha li gy tch m, cng lm cho c th kh vn ng, hot
ng chc nng kh khn v sc kho cng km i. Do vic cung cp thc n
hp l l ti quan trng.
Ch n ung b sung trong th thao gm cc thc n v ung c tc
dng ci thin nng lc vn ng ca VV. Hin nay cc dc liu th thao pht
trin rt phong ph.
Thc n b sung cho vn ng vin gm dc liu, thc phm c th lm
t ng, t cc cht in gii, t nc hoa qu, dc liu nhiu oxy. Dc liu
thc n c ngha c bit l c th gp phn nng cao thnh tch n s cung
cp nc v ng cho VV.
Ta bit rng, nng lng v s trao i cht ca c th VV lc tp luyn
ln gp 10- 20 ln so vi lc ngh ngi. Phn ln nng lng tiu hao cung cp
cho hot ng c bp, phn khc cho qu trnh chuyn ho v lm tng thn
nhit. Lng vn ng tp luyn cng ln s sinh nhit cng nhiu, thn nhit
cng cao, c th khng to nhit kp thi c v nh hng khng tt n chc
nng vn ng ca c th. Do vy, trong qu trnh vn ng, VV ra c nhiu
m hi l c th to nhit tt. 1ml m hi thot ra c th c th c tn
nhit 0,75Kcal. Do vy, cng vn ng, mi trng tp luyn cng nh vic
tiu hao nng lng rt ln ng ngha vi vic cn b sung nng lng bng
thuc hay thc phm chc nng l rt cn thit v quan trng.
Vi li th v k tc v pht huy nhng kinh nghim c truyn ca cha
ng ta li, kt hp vi s pht trin v khoa hc cng ngh hin i v tim
603

nng ngun ti nguyn sn c trong nc. Ngy nay nhiu Cng ty trong nc
cho ra i nhng sn phm c gi tr v hiu qu cao trong vic cha tr v b
dng sc kho lm phong ph thm th trng sn phm dc trong nc.
ti da trn c s kinh nghim dn gian y hc c truyn Vit Nam.
Theo ti liu ng y v ca cc nh nghin cu Tt Li cng nh ca tc gi
V Vn Chi th sao bin l mt hi sn qu ch bin mn n v lm thuc b
dng cho ngi gi, yu, mt mi v tr em chm ln. Sao bin c th ch bin
lm sch, phi hoc sy kh. C th ch bin thnh bt, n cho tht hoc ngm
ru ung. T sao bin ngi ta c th chit sut ra c cc cht kch thch min
dch, n c th kch thch nhng bin php bo v min dch ca c th, ng vai
tr ct li trong bnh hc ung th v cc bnh vim nhim. Qua iu tra c bn,
ngi ta cho bit cc loi sao bin Asterias forbesi, Acanthaster planei, Asteria
pectinifera c tnh khng sinh. Cc nghin cu iu tra phn tch gn y ca
chng ti cho thy trong mt s loi sao bin nh: Sao bin Gai lc (Comb Sea
Star - Astropecten velilaris), Sao ct Spiny (Spiny Sand Star - Astropecten vappa),
Sao Ct Gallopping (Galloping sand star - Stellaster equestris) rt giu cc hot
cht sinh hc nh cc axit amin, cc hoocmn steroit v cc nguyn t vi lng
cn thit cho c th (Kt qu c cng b trong bo co khoa hc. Nhng
vn nghin cu c bn trong khoa hc s sng 2007). Nhng kinh nghim y
hc c truyn v nhng nghin cu pht hin mi v sao bin s l c s khoa
hc nghin cu v thc hin ti.
Sao bin thng c hnh dng sao nm cnh (i khi nhiu cnh) hoc
hnh nm cnh, kch thc t 1, 2 n vi chc cm hay hn. Mt i din ca sao
bin c th c nhiu mu sc rc r. Trn c th sao bin c th phn bit ng
phng x l nhng ng i t gia c th ra tn u cnh v ng gian phng
x gia hai cnh v tn cng ra a. Sao bin b nh hot ng ca chn ng
phn di ca mi cnh. Chng di chuyn c l nh h thng ng nc cha
y dch lng, bng cch dn nc qua h thng ng nc vo chn ng v pha
chuyn vn m sao bin i c nhng tc rt chm, trung bnh khng qu 38cm trong mt pht.
Sao bin gm nhiu loi, thuc nhiu h khc nhau trong lp sao bin
Assterroidea. trn th gii sao bin c khong 1500 loi, Sao bin sinh sng
khp cc bin v i dng (tr nhng vng b ngt ho) n su ti 8500m.
b bin nc ta gp khong 5-6 loi, ph bin nht l sao bin gai lc (Comb
Sea Star - Astropecten velilaris) thuc loi h Astropectinidea. Con chong chng
Astropecten velilaris chim u th ven bin Nam Hi, vnh bc b, rt ph
bin vng y ct nh.
Nc ta c ngun tim nng v bin l v tn, Ngun sao bin phc v cho
nghin cu v sn xut l rt di do v phong ph. y cng chnh l mt trong
nhng tiu ch ti a ra t tnh kh thi.
Nhng hot cht sinh hc c trong sao bin u l nhng hot cht cn b
sung cho vn ng vin trong qu trnh tiu hao nng lng, tng cng th lc
v hi phc nhanh sc kho trong tp luyn v thi u nh (testosteron, cortisol,
st, kmv..v.).

604

Vic nghin cu thc phm chc nng tng cng th lc cho vn ng

vin s mang li nhiu hiu qu kinh t v x hi nh gim chi ph nhp khu
thuc v thc phm chc nng ngoi, an ton cho sc kho ca vn ng vin v
ch ng ngun cung cp thuc v thc phm chc nng cho vn ng vin cho
cc gii thi u trong v ngoi nc.
Kinh nghim c truyn v gi tr dinh dng thc phm thuc t mt s
loi sao bin hay t nguyn liu ng vt hay tho dc t nghn i xa cho
n nay vn cn gi nguyn c gi tr. K tha v pht trin cc sn phm trn,
ti pht trin khng ch l sn phm thc phm thng thng nh ngy xa
m vi cng ngh hin i ti cho ra i sn phm l nhng vin nang c
sn xut trn dy chuyn v cc thit b hin i. Vi sn phm ny cn mang
tnh hin i dng c cho nhiu i tng c nam gii v n gii, dng cho
nhiu la tui khc nhau.
Sn phm vin tng lc Bionamine c sn xut t nguyn liu l loi sao
bin gai lc (Comb Sea Star - Astropecten velilaris), bng cc cng ngh ho
sinh, cng ngh enzim, cng ngh ho hc..v..v... Vi nhng thnh phn dinh
dng kh phong ph c trong bt nh (Cc axit amin, cc nguyn t vi lng,
cc hot cht steroit...) cho thy ngun nguyn liu ny c nhiu hot cht gi tr
cho sc kho.
T l do cc vn nu trn, vic nghin cu sn xut vin tng lc
Bionamine nhm nng cao sc kho v tng cng th lc vn ng vin l th
hin s pht huy tim nng v ni lc v ngun ti nguyn v trnh khoa hc
cng ngh Vit Nam trong thi k cng nghip ho, hin i ho t nc nhm
phc v tt cho cc vn ng vin tham gia cc gii thi u trong nc v trong
khu vc.
X.2. kt qu v tho lun
X.2.1. IU TRA NGHIN CU V THNH PHN CC HOT CHT
SINH HC TRN 3 LOI SAO BIN - Comb Sea Star, Spiny,
Gallopping MT S VNG BIN VIT NAM
Qua iu tra nghin cu thu mu thc a chng ti thy:
- Sao ct Spiny (Spiny Sand Star - Astropecten vappa) c phn b nhiu
nht l vng bin Nha Trang Khnh Ho sau n vng bin Vng Tu
B Ra tip n l Kin Giang v Ct B - Hi Phng.
- Sao Ct Gallopping (Galloping sand star - Stellaster equestris) c phn
b nhiu nht l vng bin Kin Giang sau n vng bin Vng Tu B
Ra tip n l Nha Trang Khnh Ho v Ct B - Hi Phng.
- Sao bin Gai lc (Astropecten velilaris) c phn b nhiu nht l
vng bin Ct B - Hi Phng sau n vng bin Nha Trang Khnh Ho tip
n l Vng Tu B Ra v Kin Giang.
Cng qua cc kt qu phn tch iu tra hm lng v cc hot cht sinh
hc ca 3 loi sao bin theo s phn b c th trn chng ti thy hm lng cc
hot cht sinh hc ca ba loi sao bin cng tng cao theo s phn b v sinh

605

thi. iu ny rt c ngha trong vn chn ngun nguyn liu cho vic

nghin cu v sn xut.
Phn tch thnh phn axit amin v cc hoocmn steroit v cc nguyn t vi
lng c trong 3 loi sao bin Vit Nam: Sao bin Gai lc (Comb Sea Star Astropecten velilaris), Sao ct Spiny (Spiny Sand Star - Astropecten vappa), Sao
Ct Gallopping (Galloping sand star - Stellaster equestris), chng ti pht hin
thy trong 3 loi sao bin ny rt giu cc hot cht hoocmn steroit, cc nguyn
t vi lng.
Thnh phn cc axit amin, cc hot cht steroit v cc nguyn t vi lng
c trong 3 loi Sao bin Vit Nam nh sau:
X.2.1.1. Cc kt qu iu tra phn tch nghin cu v thnh phn axit amin
c trong cc mu nguyn liu th ca 3 loi sao bin: Sao bin Gai lc
(Comb Sea Star - Astropecten velilaris), Sao ct Spiny (Spiny Sand Star Astropecten vappa), Sao Ct Gallopping (Galloping sand star - Stellaster
equestris).
Bng X.2.1.1. Kt qu thnh phn v hm lng axit amin trung bnh c trong
mu th 3 loi sao bin (n v tnh: %g a.amin/100g mu).
Tn Axit amin
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.

Tirozin
Alanin
Acginin-Histidin
Histidin
Aspactic
Glutamic-Treonin
Treonin
Metionin+Valin
PhenylAlanin
Xerin
Glyxin
Valin
Izolxin
Xistein
Prolin
Lxin-izolxin
Lizin
4-Hydroprolin

Sao bin Gai lc Sao ct Spiny

X
1,18
3,09
4,28
0,17
2,12
3,87
0
3,76
4,78
2,17
3,85
2,89
2,98
1,82
3,87
3,76
1,76
0,89

SD
0,29
0,90
0,62
0,16
0,38
0,53
0,49
0,76
0,63
0,85
0,45
0,75
0,37
0,78
0,95
0,46
0,24

X
1,05
3,01
2,92
3,45
1,08
2,26
0
2,08
4,59
1,84
3,67
1,36
2,54
0
3,1
2,82
1,15
0,64

*Cc kt qu nghin cu vi gi tr thng k: n=30 , p 0,05

606

SD
0,26
0,74
0,81
0,70
0,31
0,56
0,53
1,00
0,36
0,51
0,35
0,64
0,83
0,80
0,28
0,10

Sao ct
Gallopping
X
SD
1,12
0,25
3,11
0,77
2,26
0,47
4,02
1,01
0,99
0,22
2,96
0,81
0
2,22
0,52
4,02
0,98
1,97
0,53
1,95
0,54
1,48
0,31
2,41
0,59
0
2,19
0,52
3,45
0,67
1,10
0,26
0,65
0,18

Hnh X.2.1.1.a. th axit amin trong loi Sao bin gai lc

(Comb Sea Star - Comb Sea Star - Astropecten velilaris)

Hnh X.2.1.1.b. th axit amin trong loi Sao ct piny

(Spiny Sand Star - Astropecten vappa)

607

Hnh X.2.1.1.c. th axit amin trong loi Sao ct Galloping

(Galloping sand star - Stellaster equestris)
Cc kt qu nghin cu trn cho thy trong 3 loi sao bin u rt giu cc
axit amin, ng vi kt qu nghin cu ca nhiu tc gi nghin cu v sinh
vt bin [330, 336, 340, 341, 342]. Kt qu iu tra phn tch ban u cho thy s
xut hin ca 17 axit amin trong loi sao bin Sao bin gai lc - Comb Sea Star
v 16 axit amin trong 2 loi sao bin (Sao ct piny, Sao ct Gallopping) chng t
ngun b sung dinh dng t thc phm sao bin rt di do v cht lng. c
bit l trong loi Sao bin gai lc hm lng axit amin rt cao, cao nht l
Phenylalanin sau n Acginin-Histidin, Glutamic-Treonin .v cui cng l
Histidin.
X.2.1.2. Cc kt qu nghin cu v cc hoocmn steroit c trong mu th
ca 3 loi sao bin: Sao bin gai lc (Comb Sea Star - Astropecten velilaris),
Sao ct piny (Spiny Sand Star - Astropecten vappa), Sao ct Galloping
(Galloping sand star - Stellaster equestris)
Cc kt qu phn tch nghin cu v thnh phn hoocmn steroit c trong
mu th ca 3 loi sao bin bng X.2.1.2. cho chng ta thy hm lng ngun
hot cht ny rt cao so vi nhiu loi ng vt qu him khc.
- Hm lng Testosteron (mg/kg vck) loi sao bin Gai lc - Comb Sea
Star l (45,029,47) sau l Sao ct Spiny (40,497,90) v ca Sao ct
Gallopping (40,383,71).
- Hm lng Cortison (mg/kg vck) loi sao bin Gai lc - Comb Sea Star
l (47,8910,16) sau l ca Sao ct Gallopping (39,566,96) v Sao ct
Gallopping (38,285,91).

608

Bng X.2.1.2. Hm lng cc hot cht steroit c trong cc mu th ca 3 loi

sao bin
Testosteron
Cortison
Progesteron
Oestradiol
Tn mu
(mg/kg cvk) (mg/kg cvk)
(mg/kg cvk) (mg/kg cvk)
X

SD

Sao ct
40,49 7,90
Gallopping
Sao bin gai lc
45,02 9,47
Comb Sea Star
Sao ct
40,38
3,71
Spiny

SD

38,28

SD

5,91 39,24

5,66

SD

35,42 7,56

47,89 10,16 47,29 10,42 87,93 17,56

39,56

6,96 38,12

6,58

34,67 7,64

* Gi tr thng k vi : (n= 30, p 0,05)

- Hm lng Progesteron (mg/kg vck) loi sao bin Gai lc - Comb Sea
Star l (47,2910,42) sau l Sao ct Gallopping (39,246,58) v ca Sao ct
Spiny (38,125,66).
- Hm lng Oestradiol (mg/kg vck) loi sao bin Gai lc - Comb Sea
Star l (87,9317,56) sau l Sao ct Gallopping (35,427,64) v ca Sao ct
Spiny (34,677,56).
Hm lng cc hoocmn steroit c trong ngun sao bin s l ngun thc
phm chc nng b sung cho c th rt tt, chc nng v tc dng ca mi loi
hot cht nh ti trnh by phn tng quan ti liu, b sung kch hot h
thng min dch, tuyn thng thn v tng cng th lc cho c th.
X.2.1.3. Cc kt qu nghin cu iu tra v thnh phn cc cht vi khong
c trong mu th thu c ca 3 loi sao bin gai lc - Comb Sea Star, Sao
ct Spiny, Sao ct Gallopping
Bng X.2.1.3. Kt qu phn tch hm lng cc vi khong c trong mu th 3 loi
sao bin Sao bin gai lc (Comb Sea Star), Sao ct piny (Spiny Sand Star Astropecten vappa), Sao ct Galloping (Galloping sand star - Stellaster equestris)
Tn mu
- Comb Sea
Star
Sao ct
Spiny
Sao ct
Gallopping

Cu
Fe
(mg/kg
Zn
(mg/kg vck)
vck)
(mg/kg vck)
41,182,44 217,2919,95 157,3930,81

Mn
Se
(mg/kg vck) (mg/kg vck)
36,275,29 67,8714,56

35,295,94

208,4833,33 147,8943,10 38,3410,00 68,9214,02

34,459,73

236,4551,82 142,2832,21 41,0212,13 62,4815,45

* Gi tr thng k vi: n=30, p 0,05

Qua kt qu nghin cu bng trn cho thy hm lng cc nguyn t vi

lng cn thit cho c th c trong mu th ca 3 loi sao bin cng kh cao so
vi cc i tng ng vt bin nghin cu trc y.
- Hm lng ng (Cu) trung bnh c nhiu nht l trong sao bin Gai
lc - Comb Sea Star (41,182,44mg/kg vck) tip theo l Sao ct Spiny
(35,295,94mg/kg vck) v ca Sao ct Gallopping (34,459,73mg/kg vck).
609

- Hm lng St (Fe) trung bnh c nhiu nht l trong Sao ct Gallopping

(236,4551,82mg/kg vck) tip theo l sao bin Gai lc - Comb Sea Star
(217,2919,95mg/kg vck) v ca Sao ct Spiny (208,4833,33mg/kg vck).
- Hm lng Km (Zn) trung bnh c nhiu nht l trong sao bin Gai
lc - Comb Sea Star (157,3930,81mg/kg vck) tip theo l Sao ct Spiny
(147,8943,10mg/kg vck) v ca Sao ct Gallopping (142,2812,13mg/kg vck).
- Hm lng Mangan(Mn) trung bnh c nhiu nht l trong Gallopping
(41,0212,13mg/kg vck) tip n l Spiny (38,3410,00mg/kg vck) v ca Gai
lc - Comb Sea Star (36,275,29mg/kg vck).
- Hm lng Selen (Se) trung bnh c trong Spiny (68,9214,02mg/kg
vck) tip theo l sao bin Gai lc - Comb Sea Star (67,8714,56mg/kg vck) v
ca Sao ct Gallopping (62,4815,45mg/kg vck).
S giu c cc hot cht nguyn t vi khong trong nguyn liu sao bin
s l ngun thc phm v ngun dc liu tt cho ngnh cng nghip ch bin
dc v cc thc phm chc nng.

Hnh X.2.1.3.a. th cc nguyn t vi lng trong Sao bin gai lc - Comb Sea
Star

Hnh X.2.1.3.b. th cc nguyn t vi lng trong Sao ct piny -Spiny Sand Star
610

Hnh X.2.1.3.c. th cc nguyn t vi lng trong Sao ct

Galloping - Galloping sand star
X.2.2. KT QU NGHIN CU V QUI TRNH TO BT NGUYN LIU
CHO CH PHM BioNAMINE.
X.2.2.1. Kt qu la chn enzym v cc iu kin thy phn
Sau khi iu tra hm lng cc hot cht sinh hc c trong 3 loi sao bin
trn, ti xc nh chn ra loi sao bin Gai lc (Comb Sea Star Astropecten velilaris) lm ngun nguyn liu v hm lng cc hot cht sinh hc
c trong sao bin Gai lc cao hn mt s loi sao bin khc.
Da trn kinh nghim y hc c truyn, kt hp vi cng ngh hin i.
ti nghin cu ng dng cng ngh ho sinh enzim nhm chit xut sao bin
ra sn phm giu cc axit amin v cc hot cht cn thit cho sc kho.
Sao bin Gai lc (Comb Sea Star - Astropecten velilaris) sau khi thu
hoch phn loi, chn lc, s ch xong ngm khong 1-2 gi, ra sch sau cho
vo xay nh v bc tip theo l dng cng ngh enzim ch bin. ti th
nghim nghin cu trn loi enzim papain. y l loi enzym thc vt c tc
dng lm mm tht, thc y nhanh qu trnh thu phn protein. Enzim papain
thu t nha cy u . Ngun enzym chng ti dng cho nghin cu do Vin
Cng ngh Sinh hc cung cp. la chn loi enzim trn vo vic x l nguyn
liu sao bin chng ti lm th nghim trong qu trnh thu phn trn c s
nghin cu d sc xut theo cc iu kin khc nhau v pH, nhit , thi gian v
hot ng c cht. Da trn kinh nghim cc kt qu nghin cu trc y v
cng da trn tnh cht v c th hot ng ca enzim ny hot ng tt nht l
pH trong khong t 6,0 8,0 do vy ti khoanh vng c tm hot ng
ca yu t pH v iu chnh cc yu t nhit , thi gian v t l c cht.

611

Bng X.2.2.1. Kt qu phn tch mu bt sao bin gai lc - Comb Sea Star sau
thu phn bng loi enzimpapain vi cc iu kin khc nhau
iu kin
iu kin
iu kin
iu kin
iu kin:
pH:6.0- to:40oC,pH:6.5 - to:40oC, pH:6.5 - to:45oC,pH:7.5 - to:45oC,
pH:7.5Tn Axit amin Tgian: 30' , T Tgian: 40' , T Tgian: 45' , T Tgian: 45' , T
to:40oC,
l c cht
l c cht
l c cht
l c cht
Tgian: 40', T
E:0,5%
E:0,5%
E:0.4%
E:0.7%
l c cht
E:0,5%

0,24
0,92

SD
0,05
0,26

3,17
4,11

SD
0,92
0,93

2,26
3,11

SD
0,60
0,89

3,05
2,12

SD
0,85
0,52

3,40
3,92

SD
0,98
0,91

1,02
2,83
3,11
0,14
1,22
3,10

0,25
0,74
0,83
0,04
0,30
0,87

0,82
3,12
4,86
2,65
4,28
5,31

0,15
0,83
1,20
0,75
1,13
1,30

3,67
3,17
3,12
2,12
1,28
4,82

0,93
0,75
0,90
0,52
0,35
1,15

0,22
1,02
0,93
4,26
0,24
2,14

0,05
0,17
0,13
0,83
0,04
0,51

3,02
3,77
3,27
3,02
4,80
2,82

0,85
0,98
0,90
0,83
1,04
0,63

0,91
3,52
0,39

0,24
1,00
0,10

1,84
3,46
4,78

0,41
1,01
1,24

1,98
3,38
0,22

0,53
0,87
0,05

2,21
0
2,00

0,59
0,49

2,12
2,01
3,12

0,61
0,50
0,88

2,67

0,72

6,12

1,23

2,01

0,56

0,10

0,02

2,21

0,57

2,29
0,22

0,61
0,06

3,92
4,80

1,07
1,26

3,22
3,84

0,80
1,11

2,14
1,92

0,53
0,39

0,18
2,87

0,05
0,75

0
2,99

0,79

3,82
3,12

0,83
0,88

5,27
3,12

1,30
0,74

0
3,01

0,79

0,24
0

0,06
-

3,01 0,86
5,12 1,40
58,31
18

2,11
0
48,7
17

0,44
-

0,92 0,24
0
26,28
15

0
0
40,77
15

1. Aspactic
2. GlutamicTreonin
3. Treonin
4. Xerin
5. Glyxin
6. Tirozin
7. Alanin
8. AcgininHistidin
9. Histidin
10. Valin
11.Metionin
+Valin
12. Pheny
lAlanin
13. Izolxin
14. Lxinizolxin
15. Lizin
16. 4-Hydro
prolin
17. Xistein
18. Prolin
a.amin TP
S lng
a.amin

1,00 0,28
0
26,57
16

* Gi tr thng k vi: n=30, p 0,05

Qua kt qu thu c bng X.2.2.1, ta thy:

- iu kin u tin (pH: 6,0, t0: 400C, Thi gian: 30', T l c cht E:
0,5%): thu phn sao bin bng enzim papain cho sn phm c thnh phn ti 16
axit amin.
- iu kin th hai (pH: 6,5, t0: 400C, Thi gian: 40',T l c cht E:
0,5%): sao bin c thu phn bng enzim papain cho sn phm l 18 axit amin.
- iu kin th ba (pH: 6,5, t0: 450C, Thi gian: 45', T l c cht E:
0,4%): thu phn sao bin bng enzim papain cho sn phm ti 17 axit amin.
- iu kin th t (pH:7,5, t0: 450C, Thi gian: 45', T l c cht E:
0,7%): thu phn sao bin kt qu cho sn phm c thnh phn l 15 axit amin.
- iu kin th nm (pH: 7,5, t0: 400C, Thi gian: 40', T l c cht E:
0,5%): thu phn sao bin bng enzim papain nhng cho cho thnh phn l 15
axit amin.
612

T cc kt qu nghin cu iu kin th hai (pH: 6,5, t0: 400C, Thi gian:

40', T l c cht E: 0,5%): thu phn sao bin bng enzim Papain cho sn phm
c thnh phn giu amine nht c ti 18 axit amin. V iu ny cho thy vic
dng enzim papain thu phn sao bin iu kin trn cho hiu sut ti u.
V ti la chn gii php iu kin nh trn ng dng vo vic sn xut
nguyn liu chnh cho vin tng lc Bionamine v papain l mt enzim c hot
tnh proteolytic rt mnh. Nc ta vng nhit i, c ngun u di do. y
l ngun cung cp papain ch yu. Papain d tch chit, bn vng, gi c hot
tnh trong thi gian di iu kin bo qun bnh thng.
Vi kt qu th nghim bng loi enzim papain trong cc iu kin khc
nhau thu phn nguyn liu sao bin nh trn cho thy enzim papain hot
ng tt v c hiu qu cao iu kin th hai tc l (pH: 6,5, t0: 400C, Thi
gian: 40', T l enzim c cht E: 0,5%): Sao bin Gai lc - Comb Sea Star
thu phn cho ra c 18 axit amin.
X.2.2.2. Kt qu nghin cu so snh v thnh phn axit amin c trong mu
bt nguyn liu sao bin - Sao bin gai lc Comb Sea Star, trc v sau khi
thu phn.
Bng X.2.2.2. Bng so snh v thnh phn axit amin c trong mu bt nguyn liu
sao bin - Sao bin gai lc Comb Sea Star, trc v sau khi thu phn.
Sao bin gai lc - Comb Sea Star
N.liu th
N.liu sau Thu phn
2,120,38
3,170,92
3,870,53
4,110,93
0
0,820,15
2,170,63
3,120,83
3,850,85
4,861,20
1,180,29
2,650,75
3,090,90
4,281,13
4,280,62
5,31 1,30
0,170,16
1,840,41
2,890,45
3,461,01
3,760,49
4,781,24
4,780,76
6,121,23
2,980,75
3,921,07
3,760,95
4,801,26
1,760,46
3,820,83
0,890,24
3,120,88
1,820,37
3,010,86
3,870,78
5,121,40
47,24
58,31
17
18

Axit amin
1. Aspactic
2. Glutamic-Treonin
3. Treonin
4. Xerin
5. Glyxin
6. Tirozin
7. Alanin
8. Acginin-Histidin
9. Histidin
10. Valin
11.Metionin+ Valin
12. Phenylalanin
13. Izolxin
14. Lxin-izolxin
15. Lizin
16. 4-Hydroprolin
17. Xistein
18.Prolin
Tng cng. a.amin ton phn
Tng s a.amin

* Cc kt qu x l bng phng php thng k tin hc cho gi tr thng k vi s mu n= 30

v p 0,05

Cc kt qu nghin cu bng trn cng cho thy tnh c hiu ca enzim

papain. Sn phm sau khi thu phn v hm lng v s lng lng axit amin
613

u tng ln r rt. iu ny cho thy kh nng xc tc rt tt ca enzim papain

trong qu trnh thu phn, cc protein sao bin c ct mch thnh cc
polypeptit, peptit v cc axit amin.
S xut hin ca 18 axit amin trong loi sao bin bin Gai lc - Comb
Sea Star chng t ngun b sung dinh dng t thc phm sao bin tht di do
v cht lng, hm lng axit amin cao nht l Prolin sau n Tirozin, 4Hydroprolin, Aspactic.v cui cng l Treonin.

Hnh X.2.2.2. th axit amin trong mu bt sau khi thu phn bng enzim
papain ca loi sao bin gai lc - Comb Sea Star
Cng t cc kt qu trn nghin cu v thu phn enzim chng ti cng
ly cc mu sn phm sau khi thu phn ca loi sao bin gai lc - Comb Sea
Star kim tra hm lng cc hot cht steroit v hm lng cc nguyn t vi
lng so snh gia mu th v mu sau khi thu phn.
X.2.2.3. Cc kt qu nghin cu v cc hoocmn steroit c trong mu bt
nguyn liu loi sao bin - Sao bin gai lc Comb Sea Star trc v sau khi
thu phn
Bng X.2.2.3. So snh hm lng cc hot cht steroit c trong mu bt nguyn
liu Sao bin gai lc - Comb Sea Star trc v sau khi thy phn.
Tn mu

Sao bin gai

lc- Comb
Sea Star

Testosteron
Cortison
Progesteron
Oestradiol
(mg/kg cvk)
(mg/kg cvk)
(mg/kg cvk)
(mg/kg cvk)
Mu
Sau
Mu
Sau
Mu
Sau
Mu
Sau
th
TP
th
TP
th
TP
th
TP
45,02 62,18 47,89 70,50 47,29 62,12 87,93
97,5
9,47 14,04 10,16 15,99 10,42 13,92 17,56 17,37

* Gi tr thng k vi: n=30, p 0,05

614

Cc kt qu phn tch nghin cu bng trn v hm lng hoocmn

steroit c trong mu bt nguyn liu loi sao bin gai lc sau khi thu phn cao
hn trc khi thu phn. iu ny cho chng ta thy cc hoocmn steroit cng
u c mt lng d tr trong v bc t bo. Trong mi t bo lun c mt cu
trc v bc bn nhng bnh thng khng o c. Trong qu trnh thu phn
enzim papain c ph v c lp v bc ca t bo lm bung cc hot cht
dinh dng cng nh cc hot cht steroit d tr bn trong. Do vy sau khi thu
phn hm lng cc hot cht steroit c ci thin tng ln. Hm lng
testosteron, cortison, progesteron v oestradiol ca loi sao bin gai lc - Comb
Sea Star c tng ln cao. Kt qu ny tng t nh nhiu cng trnh ca mt s
tc gi nghin cu [332, 336, 343, 344]. y cng l mt trong nhng ch tiu
ti a ra quyt nh chn ngun sao bin Gai lc - Comb Sea Star lm
nguyn liu chnh cho sn xut vin tng lc Bionamine.
X.2.2.4. Kt qu nghin cu v cc cht vi khong c trong mu bt nguyn
liu loi sao bin - Sao bin gai lc - Comb Sea Star trc v sau khi thu
phn.
Bng X.2.2.4. So snh hm lng cc vi khong c trong mu bt nguyn liu loi
sao bin - Comb Sea Star, trc v sau khi thu phn bng papain.
Cu
Fe
Zn
Mn
Se
mg/kg vck
mg/kg vck
mg/kg vck
mg/kg vck
mg/kg vck
Mu Sau Mu Sau TP Mu Sau TP Mu Sau TP Mu Sau TP
th
TP
th
th
th
th
Sao bin 41,18 42,00 217,29 228,93 157,39 162,60 36,27 40,10 67,87 86,80
gai lc 2,44 2,73 19,95 21,48 30,81 34,08 5,29 11,75 14,56 15,84
* Gi tr thng k vi: n=30, p 0,05
Tn
mu

Qua kt qu nghin cu bng trn cho thy hm lng cc nguyn t vi

lng cn thit cho c th c trong loi sao bin sau khi thu phn bng papain
cng c ci thin tng ln so vi trc. iu ny cho thy kh nng ca enzim
papain khng ch c tc dng phn hu cc protein thnh cc axit amin trong qu
trnh thu phn m chng cn c tc dng trong vic ph v lp cu trc v bc
t bo c cha cc cht dinh dng v hot cht d tr bn trong, y l lp cu
trc bn nhng khng o c, ch khi c tc dng ca enzim mnh nh papain
th lp v t bo b v v cc cht dinh dng v hot cht d tr bn trong n
mi b bung ra. Do vy sau khi thu phn hm lng d tr cc nguyn t vi
lng hu c cn thit trong qu trnh trao i nng lng ca t bo c trong cc
mu sao bin c tng ln. Kt qu ny tng t nh nhiu cng trnh ca mt
s tc gi nghin cu [324, 334, 335, 345-347]. Cc kt qu phn tch trn
chng minh cho thy trong loi sao bin - Comb Sea Star l mt ngun cung cp
cc nguyn t vi lng rt di do cho c th, y cng l mt ngun thc phm
chc nng v dc liu rt tt.

615

Hnh X.2.2.4. th cc nguyn t vi lng trong mu bt sau khi thu phn

bng enzim papain ca loi sao bin - Sao bin gai lc Comb Sea Star
X.2.2.5. qui trnh cng ngh sn xut to bt nguyn liu
Qua cc kt qu nghin cu v mu bt nguyn liu ca 3 loi sao bin
cho thy sau khi thu phn hm lng cc axit amin, cc nguyn t vi lng v
hm lng cc hot cht steroit c ci thin v cao hn trc khi thu phn,
c bit l cc mu trong loi sao bin - Comb Sea Star c hm lng cc hot
cht cao hn cc loi sao ct Spiny v sao ct Gallopping. Do vy ti tin
hnh chn loi sao bin Gai lc (Comb Sea Star - Astropecten velilaris) l
nguyn liu cho vic sn xut vin tng lc Bionamine
Thu phn ln 1 (iu kin to, pH, t
l E/c cht v thi gian thch hp)

- Chn lc v s ch nguyn liu

lc
Kim tra hot tnh v hm lng
cc hot cht sinh hc

Thu phn ln 2 (iu kin to, pH, t l

E/c cht v thi gian thch hp)

B sau thu
phn 1

lc

Dch chit ln 1

Dch chit ln 2
trn
Kim tra hot tnh v hm lng cc
hot cht sinh hc

Sy phun sng

Bt nguyn liu
Tinh ch

Hnh X.2.2.5. S qui trnh cng ngh sn xut to bt nguyn liu

616

X.2.3. KT QU NGHIN CU V C TNH CP TNH V BN

TRNG DIN CA BT bioNAMINE
Bt Bionamine c bo ch t sao bin bng cng ngh ho sinh enzim.
Da trn cc ti liu y hc c truyn cho thy sao bin l mt hi sn qu ch
bin mn n, ng y Tu Tnh vit Sao bin l mt thc phm thuc c gi tr
cha bnh v b dng cao. Trong sao bin c cha nhiu cc hot cht tng
cng th lc, b huyt, b thn, trng dng v knh thch min dch, c hot
cht khng sinh.
c tnh ca bt Bionamine c nh gi theo ch tiu: LD50 (liu c
gy cht mt na) v nhim c bn trng din trn ng vt thc nghim. Cc
nghin cu thc nghim v c tnh cp v bn trng din c thc hin ti
Trung tm nghin cu c hc Hc Vin Qun Y. Kt qu nghin cu xc nh
LD50 ca hot cht cha trong bt Bionamine c trnh by trong bng X.2.3.a.
Bng X.2.3.a. Bng nghin cu t l sng cht ( xc nh liu c LD50) ca
chut nht trng ung bt Bionamine (n=12)
S
TT

Mc
liu
(mg/g)

Lng
ung
(ml)

S
lng
chut
(con)

Sng/nhm
S
T l
lng
(%)

3,0

0,1

12

12

100

2
3
4
5

7,5
15,0
22,5
30,0

0,25
0,5
0,75*
1,0*

12
12
12
12

12
12
12
12

100
100
100
100

Ghi ch

Tng vn ng trong khong thi

gian 5-10 pht.
Tng vn ng chung
Tng vn ng chung
Tng vn ng (mc va phi)
Tng vn ng (mc va phi)

* Nhng mc liu c ung lm 2 ln.

Bng X.2.3.b. Bng kt qu nghin cu c tnh bn trng din ca chut nht

trng ung bt Bionamine (n=12)
L
i chng
Xc nh c tnh

SL
(chut)
25
25

Thng 1
Sng
Cht
25
0
25
0

Thng 2
Sng
Cht
25
0
25
0

Thng 3
Sng
Cht
25
0
25
0

T nhng kt qu thu c, trnh by trong bng X.2.3.a-b, chng ti c

mt s kin nhn xt sau:
Theo quy nh chung nh gi mt cch chnh xc v khch quan c
tnh ca bt k cht no u cn tun th mt s nguyn tc sau:
Chn ng vt th nghim thch hp. Cn chn nhng loi ng vt nhy
cm vi ho cht cn th. Theo quy nh kim tra c tnh cn phi th t nht
trn hai loi ng vt m mt trong hai loi phi khng thuc loi gm nhm.
Bt Bionamine s dng cho ngi nn thc nghim c tin hnh trn chut
nht trng p ng c yu cu ca nguyn tc xc nh c tnh. Thm vo ,
chut nht trng l loi ng vt gm nhm c dng ph bin trong

617

nghin cu y-sinh hc ti tt c cc c s nghin cu trong nc v trn th gii

Chnh v vy trong trng hp nghin cu c th ny c th p ng c yu cu
ca vic chn ng vt.
Cc ng vt c chn cn c s ng u v cc yu t sinh hc nh
trng lng, tnh trng sc kho, loi b cc i tng c d vt bm sinh hoc
khc thng vi cc c th khc... iu ny c nhm nghin cu thc hin
y .
Cc ng vt cn c chn v phn l mt cch ngu nhin m bo
tnh khch quan v trung thc ca vic xc nh c tnh.
S lng cc sc vt nghin cu cn ln gim bt sai s trong qu
trnh xc nh c tnh. Thng thng cc ng vt nghin cu c chia thnh
nhiu l nh c vi cc mc liu khc nhau. S lng ng vt ca mi l
v s lng l ph thuc vo tn s cc biu hin c xut hin v iu kin kinh
t, kch thc ca ng vt th nghim. Thng thng s lng l nghin cu trn
thc t phi t ti trn 10 l. S lng ng vt trong mi l theo quy nh nm
trong khong 10-20 con i vi trng hp nhim c cp tnh.
Do nh hng c mc c ca bt Bionamine l rt thp nn s
phn l c th rt bt v vi 5 l th nghim theo chng ti cng xc nh
mc c ca bt Bionamine. S lng c th trong l ca chng ti l 12
chut nm trong khong quy nh ca phng php xc nh c tnh cp v 25
chut m bo chun xc v thng k.
Thi gian theo di i vi mt cht gy c cp tnh qua ng tiu ho,
theo truyn thng, thng c xc nh l 24 gi k t khi a cht c vo c
th ng vt. iu ny cng c tun th trong qu trnh thc nghim. Cc kt
qu c th c trnh by nh sau:
Ngay t nhng liu u tin (3mg/g th trng) sau khi ung, chut bt
u c nhng bin i ban u v hnh vi. Hat ng ca chut c v nhanh nhn
hn. Chut hay chy i chy li nhiu hn so vi l i chng. Cng bt gp
nhng biu hin hng phn, tng kch thch nh. Chut hay dng li ngi, dng
tm kim.
Sau 5-10 pht hnh vi ca chut tr li trng thi bnh thng. Tuy nhin,
nu k ta thy chng nhanh nhn hn v t ng hn vo thi im bui tra
v u gi bui chiu.
So snh biu hin ca chut gia cc l th nghim chng ti thy c s
khc bit nh ch, mc hot bt ca l 4 v l 5 c v nh thp nht, c
bit l l 5.
Theo nh gi ca chng ti, s tng vn ng trong khong thi gian 510 pht u sau khi ung l hu qu ca s kch thch do ng tc ung bt buc
gy nn. Qua giai on , chut tr li mc hot ng hnh vi n nh thng
thng. iu ng quan tm l nhanh nhn, hot bt cng nh tng thi gian
thc tnh ca cc l chut c ung p buc dch hot cht cha trong bt
Bionamine cao hn cht t so vi cc l i chng. iu ny gi cho chng ta
ngh n hiu qu dc l ca bt Bionamine. Phi chng, tc dng b ca cc

618

loi axit amin v hot cht steroit cha trong bt Bionamine to ra cho ng
vt trng thi hng phn kho mnh nh gp trn.
Chng ti khng nghin cu tip qu trnh tng liu ca bt Bionamine
xc nh chnh xc liu c bi 2 l do sau:
Cho ti liu 30 mg/g th trng chut trong cc l nghin cu chng ti u
khng thy c bt k du hiu bt thng no v hnh vi.
Trn thc t liu 30 mg/g th trng l mc liu dc liu ln gp 1250 ln
mc liu c quy nh s dng cho ngi.
Nh vy cn c vo nhng kt qu thu c chng ti iu kin
kt lun rng bt Bionamine khng gy c trc ht l i vi chut v sau l
ngi bi l: mc liu 30 mg/g th trng chut, cc hot cht cha trong bt
Bionamine khng gy c bt k mt biu hin c no trn chut th nghim.
Theo tnh ton, y l mc liu cao hn 1250 ln liu c quy nh s dng trn
ngi. Trn thc t kh ai c th tip nhn c liu cao hn na bi nu theo
cch ng gi nh hin nay (300 mg/vin) c c liu 30mg/g th trng th
mt ngi nng trung bnh 50 kg phi ung s lng thuc l 5000 vin trong
mt ngy m (theo liu quy nh ch phi ung 4 vin/ngy).
Theo quy nh chung, xc nh mc c ca mt cht bt k ngi
ta phi da vo ch tiu liu gy cht ca cht . Cn c vo c cp tnh, t
chc y t th gii phn chia tt c cc ho cht thnh 5 nhm mc c khc
nhau. Cc cht thuc nhm c mc 5 (mc thp nht) c ch s LD50 nm di
5000 mg/kg. Theo cch nh gi ny th nhng cht c LD50 nm trn mc 5000
mg/kg th trng s c xem l cht khng c. Trong thc nghim ca chng
ti bt Bionamine s dng ti mc liu 30.000 mg/kg th trng v khng gy
t vong mt c th no trong l th nghim. Nh vy LD50 ca bt Bionamine
chc cn cao hn mc liu 30000 mg/kg th trng nn c th khng nh rng bt
Bionamine khng c.
Thc nghim bn trng din (kt qu trong bng X.2.3.b) cho thy bt
Bionamine khng gy cht ng vt th nghim trong khong thi gian s dng
03 thng lin tc mc liu 15.000 mg/kg th trng (gp 625 ln s dng cho
ngi) khi xm nhp vo c th qua ng tiu ho.
* Kt lun
1. Bt Bionamine liu 30000 mg/kg th trng khng gy c i vi chut
th nghim. Khng lm cht chut hoc gy xut hin bt k triu chng nhim
c no.
2. Vic xc nh chnh xc LD50 ca bt Bionamine trn thc t khng cn
thit v chc chn s cao hn mc liu 30.000 mg/kg th trng nhiu ln, m
thm ch ngay mc liu ny th mt ngi c trng lng trung bnh 50 kg cng
phi tip nhn trong vng 24 gi mt s lng 5000 vin Bionamine. iu ny
kh c kh nng xy ra trong thc t.
3. Bt Bionamine khng gy nh hng bn trng din trn ng vt thc
nghim ngay c khi s dng liu cao, gp 625 ln so vi liu dng cho ngi,
trong thi gian di 3 thng.

619

X.2.4. KT QU NGHIN CU V QUI TRNH TO VIN TNG LC

BIONAMINE
X.2.4.1. Nghin cu to vin tng lc Bionamine
Da trn kinh nghim v cc bi thuc y hc c truyn, sn phm bt sao
bin Gai lc (Comb Sea Star - Astropecten velilaris) sau khi thu phn l
nguyn liu ca vin tng lc Bionamine c phi ch vi cc ph gia, t dc
theo cng thc v thnh phn nh sau:
- Thnh phn hn hp nguyn liu gm bt sao bin Gai lc thu phn,
nhn sm, vitamin B1, B2 v t dc va , trn u. To m cm bng h tinh bt
sy kh. Trn tip vi lng va Talc magnesi stearat c cm.
- em cm ng nang vi khi lng nht nh t hm lng nguyn
liu theo tiu chun (Sai s cho php 64010% mg/vin) vo vin nang, ng
10vin/v/, 1 hp/4v.
X.2.4.2. S quy trnh to vin tng lc Bionamine
Lactose, tinh bt

Talc, Mg stearat

Hn hp nguyn liu

To bt kp, sy kh

Cm kh

ng gi

p v

ng nang

Hnh X.2.4.2.a. S qui trnh cng ngh sn xut vin tng lc Bionamine
Sn phm nguyn liu v vin tng lc Bionamine c sn xut trn
dy chuyn thit b hin i t tiu chun ti Cng ty Cng ngh Intecpharm.

Hnh X.2.4.2.b Sn xut vin tng lc Bionamine trn dy chuyn thit b

hin i ti Cng ty Cng ngh Intechpharm

620

Sn phm c xy dng tiu chun c s v c B Y t cp Chng

nhn tiu chun sn phm s 956/2008/YT-CNTC vi tn gi Thc phm chc
nng: Vin tng lc Bionamine.

Hnh X.2.4.2.c. Hnh nh sn phm v chng nhn tiu chun ca B Y t

621

X.2.5. KT QU NGHIN CU thnh phn v tc dng ca VIN

TNG LC BIONAMINE trn ng vt
Da trn cc bi thuc y hc c truyn, Vin Bionamine vi thnh phn
chnh l cc hot cht sinh dc c chit xut t nguyn liu sao bin c
phi ch vi cc dc liu, ph gia, t dc, vi thnh phn l nhng nguyn liu
c cha nhiu cc protein, axit amin, cc vitamine trong nhm B, cc hot cht
steroit v cc cht vi khong cn thit cho c th, c ng vo nang s 0 vi
hm lng Bionamine l 300 mg. Sn phm vin nang tng lc Bionamine c
sn xut trn dy chuyn thit b t tiu chun Cng ty TNHH Cng ngh
Intechpharm, sn phm ng mi v l 10vin/v, 2v/ti v 2ti/hp,bao b, cht
lng t cc tiu chun c B Y t cho php, giy php s 956/2008/YTCNTC. nh gi tc dng ca vin tng lc Bionamine trong vic hi phc sc
kho v tng cng th lc ca vn ng vin. ti tin hnh nghin cu v
thu c mt s kt qu nghin cu nh sau:
X.2.5.1. Kt qu nghin cu v thnh phn axit amin vin tng lc
Bionamine
Qua kt qu nghin cu trong bng X.2.5.1. cho chng ta thy hm lng
cc axit amin c trong sn phm vin tng lc Bionamine so vi cc sn phm
khc trong nc nh Phunamine, Taxaton, ru Hi m th vin tng lc
Bionamine c gi tr cao hn v cng khng thua km g so vi sn phm vin
Triplex hay gi khang thai ca Trung Quc. sn phm gi Khang thai, thnh
phn c 17 axit amin v hm lng cng tng i cao nh: Histidin: 9,88
Xistein: 9,54, Lxin: 8,21, A,Aspactic: 7,22, Trong khi vin tng lc
Bionamine thnh phn cng c ti 18 axit amin v hm lng cng rt phong ph
nh: A,Aspactic: 3,18, A,glutamic: 4,21, Treonin: 2,92, Xerin: 3,26, Glyxin:
6,85, Tirozin: 3,56, Alanin: 5,68, Acginin: 5,71, Histidin: 3,74, Valin: 3,16,
Metionin: 4,38, Phenylalanin: 7,42, Izolxin:3,98, Lxin: 4,80, Lizin: 3,82, 4Hydroprolin: 2,12, Xistein+Xistin: 3,06, Prolin: 5,12, kt qu trn ph hp vi
nhiu tc gi nghin cu trc y [334, 348, 349].

Hnh X.2.5.1. th axit amin c trong vin tng lc Bionamine

622

Bng X.2.5.1. Hm lng cc axit amin trong vin Bionamine so vi cc sn

phm khc (n v tnh:%g a.amin/100g mu).
Tn axit amin

1. A.Aspactic
2. A.glutamic
3. Treonin
4. Xerin
5. Glyxin
6. Tirozin
7. Alanin
8. Acginin
9. Histidin
10. Valin
11. Metionin
12. PhenylAlanin
13. Izolxin
14. Lxin
15. Lizin
16. 4-Hydroprolin
17. Xistein+Xistin
18. Prolin

Vin
Bionamine

Vin
Phunamine

Ru
Hi
M
(VN)

Vin
Taxaton
(VN)

Gi
Khang
Thai
(TQ)

Vin
Triplex
(TQ)

3,18
4,21
2,92
3,26
6,85
3,56
5,68
5,71
3,74
3,16
4,38
7,42
3,98
4,80
3,82
2,12
3,06
5,12

7,20
4,80
2,04
3,01
3,28
10,45
3,18
7,19
6,10
3,05
4,60
8,04
7,20
3,20
6,02
7,62
3,00
10,42

3,24
2,50
0,12
1,12
3,70
0,38
0,52
1,48
1,26
2,01
1,18
1,05
2,01
1,53
0,85

3,03
2,80
1,22
4,68
6,96
7,50
3,50
3,10
3,87
7,80
2,82
2,90
4,85
2,55
7,20
5,80
6,50
0,12

7,22
6,15
4,02
3,11
6,28
4,02
4,07
5,78
9,88
4,20
6,19
5,44
3,19
8,21
3,15
7,14
9,54
7,89

4,12
6,05
2,80
2,18
10,88
2,90
5,92
4,03
3,99
1,80
1,49
3,49
2,77
3,12
4,28
3,60
4,82

Chng ta u bit cc axit amin ng vai tr rt quan trng v thit yu

trong trong c th. Mi axit amin u c nhng ng gp tng chc nng cho c
th nh: A.Aspactic l axit amin c tc dng gip cho qu trnh gii c v tng
cng sc bn c th. A.glutamic b no, gip mau lnh vt thng, kch thch
hng phn thn kinh. Treonin kch thch h thng thn kinh thc vt gip cho ng
ngon tng cng chc nng gan. Xerin tng cng min dch, sc khng v
tng sc bn. Glyxin tng c bp, gip tuyn tin lit kho mnh.
Tirozin tng cng sinh lc, hi phc sc kho, b gan. Alanin n nh
ng trong mu. Acginin l axit amin thit yu c tc dng tng cng h thng
min dch, tng trng hoocmn gip cho ngn nga v iu chnh cc chng bt
lc ca nam gii.
Histidin c tc dng b khp, tng to mu chng d ng. Metionin, Valin
gip cho n nh tm l. PhenylAlanin c tc dngngn nga chng bo ph,
gim trm cm, gim au nhc do bnh cm, kinh nguyt, gim vim khp.
Izolxin l axit amin thit yu n nh lng ng trong mu, tng cng h tr
pht trin c bp.
Lxin lm tng c bp, n nh tm l. Lizin gip cho qu trnh hnh thnh
khng th, n nh ni tit t. Hydroprolin, Xistein tng cng min dch chng
xy ho, v hiu ho cc gc t do, gip cho mau lnh vt thng, pht trin v
lm p t bo da, tc. Prolin c tc dng gip cho qu trnh vn ng khp v
cc c chng, c tim [330, 331, 337, 338].

623

Vi thnh phn axit amin ca vin tng lc Bionamine so vi cc sn

phm tng t khc trong nc v nc ngoi khng km g v gi tr chc
nng dinh dng do vy sn phm rt c tin dng cho vic nng cao th lc ca
vn ng vin l rt tt [324, 335, 336, 350].
X.2.5.2. Nghin cu thnh phn cc cht vi khong trong vin tng lc
Bionamine
V hm lng cc hot cht vi khong sau khi bo ch vin tng lc
Bionamine qua kt qu nghin cu trong bng X.2.5.2. cho thy hm lng cc
cht vi khong c trong sn phm vin tng lc Bionamine so vi cc sn phm
tng t trong nc cao hn v so vi sn phm gi khang thai v vin Triplex
ca Trung Quc th kt qu cng khng khc nhau nhiu nh ca Bionamine,
thm ch mt s thnh phn ca hm lng nguyn t v st, km hay selen cn
cao bng hoc hn cc sn phm ni v ngoi, nhng kt qu ny ng vi nhiu
tc gi nghin cu trc y [331, 335, 349, 351].
Bng X.2.5.2. Hm lng cc cht vi khong c trong vin Bionamine so vi cc
sn phm khc.
Hm lng vi
cht
Cu(mg/kg vck)
Fe (mg/kg vck)
Zn (mg/kg vck)
Mn(mg/kg vck)
Se(mg/kg vck)

Vin
Bionamine

Vin
Phunamine

4,20
228,93
16,26
40,10
86,80

3,22
231,08
7,27
32,90
-

Vin
Taxaton
(VN)
4,01
227,20
6,26
27,24
-

Gi
Khang
thai (TQ)
6,22
240,02
11,28
37,91
-

Vin
Triplex
ca (TQ)
3,92
238,27
3,24
42,76
-

Hnh X.2.5.2. Biu hm lng cc cht vi khong c trong

vin tng lc Bionamine
Vi thnh phn cc cht vi khong hu c c trong vin tng lc
Bionamine rt phong ph c bit l nguyn t st s gip cho qu trnh tng to
hng cu, hemoglobin trong c th rt tt. Hm lng ng, mangan c trong

624

vin Bionamine gip cho s pht trin tr tu, kch thch phn x thn kinh c
tt hn, hm lng km c trong vin Bionamine cao s gip cho qu trnh tng
cng th lc, tng cng min dch gip c th thc hin tt chc nng ni tit,
tun hon mu, mau lnh vt thng, khng ch t bo gy nhiu. c bit
nguyn t km cn ng vai tr chc nng duy tr sinh l ho trong c th, tng
kh nng sinh dc cho nam gii, gim bt cc nguyn t vi lng t do. kch
thch cc hoocmn tuyn thng thn hot ng [328, 332, 337].
X.2.5.3. Nghin cu thnh phn cc hot cht hoocmn steroit trong vin
tng lc Bionamine
V hm lng cc hot cht steroit, sau khi bo ch vin tng lc
Bionamine Qua kt qu nghin cu trong bng X.2.5.3 cho thy hm lng cc
hot cht steroit c trong sn phm vin tng lc Bionamine cng kh cao so vi
cc sn phm khc, k c sn phm ngoi, iu ny chng t sn phm trong
nc nu c ng dng khoa hc cng ngh th cht lng v gi tr s c
nng ln r rt. Hm lng Testosteron (mg/kg vck) c trong vin Bionamine l
(69,02) cao hn vin Taxaton (68,00) v gi khang thai ca Trung quc (67,35)
nhng cao hn vin Triplex ca Trung quc (32,08).
Bng X.2.5.3. Hm lng cc hot cht steroit c trong vin Bionamine so vi cc
sn phm khc.
Hm lng steroit

Vin
Bionamine

- Testosteron(mg/kg vck)
- Progesteron(mg/kg vck)
- Cortisol (mg/kg vck)

69,02
79,73
56,02

Ru
Hi m
(VN)
11,02
90,28
8,22

Vin
Gi
Vin
Taxaton
Khang
Triplex
(VN)
thai (TQ) ca (TQ)
68,00
67,35
32,08
65,07
52,15
38,72
19,40
59,24
55,12

Hm lng Progesteron(mg/kg vck) ca vin Bionamine l (79,73) cao

hn nhiu so vi cc sn phm trong nc v ca Trung Quc nh ru Hi m
(9,28), vin Taxaton (65,07), gi khang thai (52,15) v vin Triplex (38,72).
Hm lng Cortisol (mg/kg vck) ca vin Bionamine l (56,02) cao hn tt
c cc sn phm trong nc nh ca ru Hi m (8,22), vin Taxaton (19,40) v
khng thua km g cc sn phm ca Trung Quc nh gi Khang Thai (19,40),
vin Triplex (55,12).
Vi thnh phn v hm lng cc hot cht hoocmn steroit c trong vin
tng lc Bionamine c tc dng ln cc chuyn ho trung gian, Gluxit, lipit v
protein. Cc hoocmn steroit kch thch tng hp lipoprotein gip cho vic b p
v cn bng glucose v lipit trong mu. Hoocmn steroit vi nng cao c tc
dng tng cng min dch, chng vim nhim, chuyn ho mui ln tuyn nc
bt v tuyn m hi cng nh s vn chuyn mui v nc qua nim mc ng
tiu ho, tham gia vo s duy tr cn bng nc v in gii m bo s vn ng
tiu hao nng lng c th [331, 334, 337, 345, 352].
Thnh phn hot cht hoocmn steroit c trong vin Bionamine s lm
tng gi tr b dng. Chnh v vy vic chn vin tng lc Bionamine dng cho
vn ng vin l rt cn thit v quan trng. Trc khi dng th nghim chnh
625

thc cho vn ng vin ti c lm thc nghim v tc dng tng cng th lc

trn ng vt v thu c cc kt qu nh trnh by di y.
X.2.5.4. Kt qu nghin cu v th nghim tc dng ca vin tng lc
Bionamine trn ng vt thc nghim.
a. Tc dng ca vin tng lc Bionamine ln mt s ch tiu huyt hc ca
chut cng trng.
Bng X.2.5.4.a. Bng kt qu kim tra cc ch tiu huyt hc ca chut dng th
nghim ch phm.
L Th nghim dng Vin Bionamine
Tn
CT

WBC
(x103/l) S
lng tuyt
i bch cu
oan trung
tnh

RBC
x106/l)
S lng
hng cu

HGB
(g/dL)
Huyt sc t

Trc Sau Trc Sau Trc

2,30 2,00 3,33 4,04 12,76
SD 0,22 0,22 0,48 0,67 0,55
* Gi tr thng k vi: n = 10, p 0,05
X

Sau
14,02
0,84

L i chng dng Vin Placebo

WBC
(x103/l)
RBC
HGB(g/dL)
S lng
Huyt sc
(x106/l)
tuyt i
t
S lng
bch cu
hng cu
oan trung
tnh

Trc Sau Trc Sau Trc Sau

2,00
0,18

2,10
0,16

3,34
0,43

3,39 12,78 12,67

0,46 0,73 0,68

Kt qu bng trn cho thy sau khi cho chut cng trng n b sung vin
tng lc Bionamine trong 20 ngy th lm thay i mt s ch tiu huyt hc
quan trng ca chut.
S lng hng cu (RBC) trung bnh ca chut nhm (TN1) cho n b
sung ch phm Bionamine trc th nghim l 3,33 (x106/l), sau th nghim l
4,04 (x106/l), Tng 0,91 (x106/l). Trong khi nhm i chng (C2) cho
n vin Bionamine gi (Placebo) trc th nghim l 3,34 (x106/l), sau th
nghim l 3,39 (x106/l). Tng 0,02 (x106/l), ch tiu ny cho thy hng cu
l i chng trc v sau th nghim c tng khng ng k.
S lng Hemoglobin (HGB) trung bnh ca chut l (TN1) cho n vin
tng lc Bionamine trc th nghim l 12,76 (g/dl), sau th nghim l 14,02
(g/dl). Tng 1,26 (g/dl). l (C2) cho n vin Bionamine gi (Placebo) trc
th nghim l 12,78 (g/dl), sau th nghim l 12,67 (g/dl) gim 0,11 (g/dl).
S lng bch cu (WBC) trung bnh ca chut cc l th nghim c
thay i nhng khng ng k. l th nghim (TN1) cho n b sung vin tng
lc Bionamine trc th nghim l 2,3 (x103/l), sau th nghim l 2,0 (x103/l).
l (C2) cho n vin Bionamine gi (Placebo) trc th nghim l 2,0
(x103/l), sau th nghim l 2,10. S tng gim ca bch cu c l i chng v
th ngim l khng c ngha.
Qua kt qu th nghim trn cho thy vin tng lc Bionamine thc s c
tc dng kch thch s to mu, to hng cu v hemoglobin. S tng s lng
hng cu c ngha tng cng cung cp oxy cho t bo, iu rt cn thit cho
qu trnh vn ng v cc hot ng ca c bp cng nh vic tng cng th lc
ca c th.
626

b. Tc dng tng cng th lc v sc bn ca ch phm Bionamine trn

chut trong qu trnh bi ln.
Bng X.2.5.4.b. Bng kt qu kim tra thi gian bi ca chut.
Chut TN

Chut ci
Chut c
X

L i chng
TrcTN
Sau (TN) 15ngy

3,36
3,25
3,31

3,41
3,35
3,38

L (TN1)B sung Bionamine

TrcTN
Sau (TN) 15 ngy

3,26
3,28
3,27

4,07
4,02
4,05

* Gi tr thng k vi: n = 10, p 0,05

Qua kt qu bng trn cho thy gia cc l i chng v th nghim c s

khc nhau r rt, thm ch s khc nhau cn th hin c kt qu trong cc l
th nghim t i chng.
- l i chng thi gian bi trung bnh ca chut gia trc th nghim l 3,31
v sau t th nghim l 3,38 kt qu c s khc nhau khng nhiu lm.
- l th nghim (TN) chut th nghim c cho n b sung ch phm
Bionamine v thi gian bi trung bnh c ci thin tng c 0,38 trc th
nghim l 3,27 , sau th nghim kt qu l 4,05 .
Cc kt qu thc nghim trn chng minh cho thy cc ch phm
Bionamine thc s c tc dng tng cng th lc i vi chut th nghim th
hin thi gian bi ca chut c ko di hn, sc chu ng ca chut c tt
hn. iu ny l gii rng trong cc ch phm cung cp b sung thm cc cht
dinh dng cng nh cc hot cht sinh hc, cc hoocmn steroit (Testosteron,
progesteron), cc cht vi khong nh Fe, Zn, Cu..v cc axit amin cn thit
trong qu trnh vn ng v tiu hao nng lng ca chut.
T cc thc nghim chng minh c tc dng ca vin tng lc
Bionamine i vi sc bn v tng cng th lc trn chut, ti tin hnh
trin khai thc nghim trn vn ng vin mn in kinh, bng bn, mn c t
v thu c cc kt qu di y.
X.2.6. Kt qu nghin cu v th nghim vin tng lc
Bionamine trn vn ng vin
X.2.6.1. nh gi v tc dng sc bn trn vn ng vin
nh gi c sc bn ca vn ng vin in kinh ti tin hnh
ly mu mu v kim tra cc ch tiu sinh ho lin quan n sc bn ca th lc
vn ng vin v kt qu cho thy:
a. Tc dng hi phc v nng cao hm lng Testosteron trong mu vn ng
vin ca vin tng lc Bionamine.
Kt qu bng X.2.6.1.a cho thy hm lng testosteron ca VV
nhm i chng (khng dng vin Bionamine) sau 1 thng tp luyn v thi u:
VV nam gim 3,62% t 24,28 nmol/l xung 23,40 nmol/l, cn VV n
gim 11,93%, t 2,18 nmol/l xung cn 1,92 nmol/l.

627

Bng X.2.6.1.a. Hm lng Testossterone trong mu ca VV in kinh trc v

sau 1 thng luyn tp.
Hm lng
Testossterone (nmol/l)

Nam VV

SD
N VV

SD

Nhm C dng placebo

Sau 30 ngy
Trc t tp
tp

24,28
3,71
2,18
0,49

23,40
3,84
1,92
0,26

Nhm TN dng Bionamine

Trc t
Sau 30 ngy
tp

21,92
1,92
1,46
0,44

24,95
1,83
2,04
0,39

* Gi tr thng k vi: n = 14, p 0,05

nhm th nghim dng vin tng lc Bionamine sau 1 thng tp luyn

v thi u hm lng testosteron nh sau: Vn ng vin nam trc khi dng v
sau khi dng tng 13,84%, t 21,92 nmol/l n 24,95 nmol/l. Nhm vn ng
vin n sau 1 thng dng vin tng lc Bionamine hm lng testosteron tng
t 1,46 nmol/l ln 2,04nmol/l.
Cc kt qu trn cho thy rng vin tng lc Bionamine gp phn ci thin
hm lng testosterrone hao ht t nhin trong c th sau t tp luyn, gip cho
vn ng vin hi phc nhanh hn. iu ny ng vi kt qu nghin cu ca
nhiu tc gi [331, 335].
b. Tc dng b sung b p nng cao hm lng cortisol trong mu vn ng
vin tc ng tch cc vo qu trnh nng cao th lc v hi phc sc kho cho
vn ng vin ca vin tng lc Bionamine.
Cortisol (l hoocmn thuyn thng thn) v l mt tham s quan trng
trong vic nh gi lng vn ng ca c th. Khi c th vn ng nng s sinh
ra s tiu hao nng lng ln
Kt qu bng X.2.6.1.b. cho thy hm lng cortisol trong mu gia cc
nhm i chng v nhm nghin cu dng vin tng lc Bionamine c s khc
nhau r rt.
Bng X.2.6.1.b. Hm lng Cortisol trong mu ca VV in kinh trc v sau 1
thng luyn tp
Hm lng Cortisol
(nmol/l)

Nam VV
N VV

Nhm C dng placebo

Nhm TN dng Bionamine

Trc t
Sau 30 ngy
tp
541,55
584,42

Trc t tp

Sau 30 ngy tp

545,93

536,55

SD

57,12

52,69

51,05

64,07

542,25

530,25

540,17

544,00

SD

78,05

72,15

75,46

96,53

* Gi tr thng k vi: n = 14, p 0,05

- Vn ng vin n nhm i chng sau mt thng tp luyn khng

dng b sung vin Bionamine hm lng cortisol b hao ht t 542,25 (nmol/l)
xung cn 530,25 (nmol/l). Trong khi nhm nghin cu c dng b sung
vin tng lc Bionamine sau 1 thng tp luyn hm lng cortisol trong mu

628

khng nhng khng b gim m cn c b p tng thm t 540,17 (nmol/l)

ln 544,00 (nmol/l).
- Vn ng vin nam nhm i chng sau mt thng tp luyn khng
dng b sung vin Bionamine hm lng cortisol cung b gim t 545,93 (nmol/l)
xung cn 536,55 (nmol/l). Trong khi nhm nghin cu c dng b sung
vin tng lc Bionamine sau 1 thng tp luyn hm lng cortisol trong mu
tng thm t 541,55 (nmol/l) ln 584,42 (nmol/l). Cc s liu so snh trn u
mc cho php p0,05 do vy kt qu nghin cu l hon ton c gi tr.
T vn hm lng cortisol t nhin c trong thnh phn ca vin tng
lc Bionamine cao. Thng qua vic dng b sung vin tng lc Bionamine cho
vn ng vin in kinh nghin cu trn chng minh rng vin tng lc
Bionamine b sung rt tt v kp thi vo qu trnh hao ht cortisol (l
hoocmn tuyn thng thn) gim st trong qu trnh vn ng. Khc vi
nhng hoocmn protein, cortisol cng nh testosteron l nhng hoocmn steroit
khng c d tr trong nhng nang bi tit v cng khng c hoocmn d tr
trong nang t bo. Cortisol cng nh testosteron c th khuch tn t do qua
mng, t bo v vo mu. trong tun hon mu, phn ln cortisol cng nh
testosteron v cc hoocmn steroit kt hp vi protein c hiu ca huyt tng,
bng cch ny cortisol cng nh cc hoocmn steroit khc tn ti t do trong
tun hon lu hn so vi dng steroit t do. Ch c cc hoocmn steroit t do l
c tc dng hot ng sinh hc. Vic b sung cortisol cng nh testosteron v
cc hoocmn steroit bng vic dng vin tng lc Bionamine c tc dng rt tt
trong vic tng cng th lc v hi phc sc kho ca vn ng vin. iu ny
ph hp vi nhiu tc gi nghin cu trc y [330, 331, 333, 352].
c. Tc dng b tr chc nng gan trong c th, kch hot enzym Lactat
Dehydrogenaza (LDH) gip tng nhanh qu trnh hi phc ca vin tng lc
Bionamine.
Qua kt qu bng X3.6.1.c cho ta thy vn ng vin cc nhm i
chng sau 1 thng tp luyn khng dng vin tng lc Bionamine hot LDH
trong huyt thanh hu nh khng thay i hoc tng ln khng ng k t 0,614,52%. vn ng vin n thay i trc tp luyn v sau tp luyn t 398,40
UI/l ln 416,40 UI/l (tng 4,52%). Cn vn ng vin nam thay i trc tp
luyn v sau tp luyn t 390,60 UI/l ln 393,00 UI/l (tng 0,61%).
Bng X.2.6.1.c. Hm lng Lactat Dehydrogenaza (LDH) trong mu ca VV
in kinh trc v sau 1 thng tp luyn.
Nhm C dng Placebo
Trc t
Sau 30 ngy
tp luyn
tp luyn

Hm lng LDH
trung bnh (UI/l)

Nam VV

SD
N VV

SD

390,60
33,11
398,40
21,30

393,00
65,15
416,40
40,27

* Cc gi tr thng k vi n=14 v p0,05

629

Nhm TN dng Bionamine

Trc t
Sau 30 ngy
tp luyn
tp luyn

428,47
36,93
381,13
43,32

478,27
53,52
425,47
31,97

Trong khi nhm thc nghim c dng b sung vin tng lc

Bionamine trong thi gian tp luyn, hot LDH nam VV tng t 428,47
UI/l n 478,27 UI/l (11,62%), n VV tng 11,63% (t 381,13 UI/l n
425,47 UI/l), iu ny c th gii thch l do vin Bionamine c thnh phn v
hm lng km hu c cao. Trong nhng cng trnh nghin cu mi gn y
khng nh rng Zn km c tc dng kch hot enzym LDH trong phn ng kh
axit lactic to thnh axit piruvic, y l mt yu t rt cn thit i vi cc vn
ng vin v cc i tng lao ng c bp. iu ny cng ph hp vi kt qu
nghin cu ca nhiu tc gi trc y [333, 335, 345, 352].
d. Tc dng trong qu trnh phn gii axit lactic trong c lm tng cng kh
nng hi phc ca c th vn ng vin ca vin tng lc Bionamine.
Bng X.2.6.1.d. Hm lng axit lactic trong mu ca VV in kinh trc v sau
1 thng tp luyn.
Hm lng axit
lactic trung bnh
(mg/dl)

Nam VV

SD
N VV

SD

Nhm C dng placebo

Sau 30 ngy
Trc t tp
tp

4,26
0,34
3,96
0,18

4,49
0,30
4,29
0,24

Nhm TN dng Bionamine

Sau 30 ngy
Trc t tp
tp

3,24
0,40
4,02
0,21

2,60
0,25
2,74
0,50

* Cc gi tr thng k vi n=14 v p0,05

Cc kt qu trong bng X.2.6.1.d cho thy cc vn ng vin nhm i

chng khng dng vin tng lc Bionamine, hm lng axit lactic nam VV
sau t tp luyn khng c g thay i m c v tng nh, tng t 4,26 mg/dl ln
4,49 mg/dl (5,4%), cn n vn ng vin nhm i chng khng dng
Bionamine hm lng axit lactic trung bnh tng t 3,96 mg/dl ln 4,29 mg/dl
(8,28%). iu ny cng rt c l v sau mi t tp luyn vn ng vin thng
mt mi, nguyn nhn gy mt mi chnh l Axit lactic tn ng li nhiu
trong c, nu c xc tc nhiu ca enzim LDH th axit lactic se c phn hu v
s lm gim mt mi, khng b co c.
Cc vn ng vin nhm th nghim c dng vin tng lc
Bionamine, hm lng axit lactic trung bnh nam VV sau t tp luyn c
ci thin gim t 3,24 mg/dl xung 2,60 mg/dl (19,63%), n VV nhm th
nghim gim 31,83% (t 4,02 mg/dl xung 2,74 mg/dl) cc kt qu ny cho thy
vin Bionamine c tc dng kch hot enzym LDH tng cng phn ng kh axit
lactic to thnh axit piruvic, y l mt yu t rt cn thit i vi cc vn ng
vin v cc i tng lao ng c bp. Kt qu ny ph hp vi nhiu tc gi
[327, 331, 337].

630

e. Tc dng hi phc v tng hng cu v ci thin cc ch tiu huyt hc ca

vn ng vin ca vin Bionamine.
Bng X.2.6.1.e. Kt qu kim tra cc ch tiu huyt hc vn ng vin in kinh
trc v sau mt thng tp luyn.
Ch tiu

Nhm VV
VV (Nam)
Nhm TN
X

SD
VV (Nam)
Nhm C
X

SD
VV (N)
Nhm TN
X

SD
VV (N)
Nhm C
X

WBC (x103/l)
S lng tuyt
i bch cu
oan trung
tnh
T
S

RBC (x106/l)
S lng hng
cu

HGB
g/dL
Huyt sc t

MCV
Th tch trung
bnh hng cu
(f/l)

6,57
1,29

6,29
1,12

4,98
0,36

5,56
0,51

14,00
0,53

14,69
0,95

88,05
4,73

89,13
4,15

6,92
1,12

7,02
0,95

4,82
0,39

4,62
0,52

13,76
1,13

13,32
1,12

87,64
2,90

87,06
2,58

6,13
0,81

5,91
0,83

4,29
0,22

4,90
0,48

12,21
0,84

12,86
0,96

88,68
2,92

90,10
3,94

6,72

6,83

4,39

4,26
0,30

12,26
1,15

11,76
1,16

87,06
3,24

86,84
2,94

SD
1,43
1,51
0,45
* Cc gi tr thng k vi n=14 v p0,05

Qua kt qu bng X.2.6.1e. chng ta thy th tch trung bnh hng cu

(MCV) ca cc VV nhm th nghim sau khi dng vin tng lc Bionamine
mc n nh tt: nam VV t 88,05 f/l n 89,13 f/l v n VV t 88,68 f/l
n 90,10 f/l.
S lng bch cu WBC ca VV nhm th nghim trc v sau t tp
luyn c dao ng nhng mc n nh tt: Nam VV t 6,57(x103/l) n 6,29
(x103/l) v n VV t 6,13 (x103/l) n 5,91 (x103/l).
Huyt sc t (HGB) cc VV th nghim u gi c mc n nh:
nam VV t 14,00 g/dl n 14,69 g/dl v n VV t 12,21g/dl n 12,86 g/dl.
S lng hng cu nhm VV th nghim sau 1 thng tp luyn c dng
vin tng lc Bionamine cho thy s lng hng cu khng nhng khng b gim
m cn c ci thin tng ln, nam VV hng cu tng 4,98 (x106/l) ln
5,56 (x106/l), n VV hng cu tng t 4,29 (x106/l) ln 4,90 (x106/l).
Cc kt qu tng t nhn c trong thc nghim vi cc VV nhm
i chng khng dng vin tng lc Bionamine trong t tp luyn nh sau:
Hng cu nam VV c thay i khng nhng khng tng m cn gim nh t
4,82 (x106/l) n 4,62 (x106/l) v n VV tng t 4,39 (x106/l) n 4,26
(x106/l).

631

Huyt sc t (HBG) nam VV nhm i chng gim t 13,76 g/dL n

13.32 g/dL. n VV gim t 12,26 g/dL n 11,76 g/dL.
S lng bch cu WBC ca VV nhm i chng trc v sau t tp
luyn c dao ng theo nh hng ca s lng hng cu: Nam VV t
6,92x103/l n 7,02x103/l v n VV t 6,72x103/l n 6,83x103/l.
Th tch trung bnh hng cu (MCV) ca cc VV nhm VV i chng
khng dng vin tng lc Bionamine sau t tp luyn hu nh khng c s thay
i: nam VV t 87,64 f/l n 87,06 f/l v n VV t 87,06 n 86,84 f/l.
f. Tc dng hiu qu ln cc test gng sc ti a ca VV ca vn ng vin
in kinh ca vin tng lc Bionamine.
Thc nghim c tin hnh kim tra nh gi trn cc vn ng vin in
kinh trc v sau 1 thng tp luyn c dng th nghim vin tng lc Bionamine.
Kt qu thu c trn thit b chn on chc nng Cosmed ca Italia cho 20
VV in kinh (10 nam v 10 n) theo bng di y.
Bng X.2.6.1.f. Cc ch s chc nng trung bnh ca vn ng vin nam in kinh
H ni trong test gng sc ti a. (Cc gi tr thng k vi n=14 v p0,05)
Ch tiu

Cng sut vn
Sc bn a kh
Chc nng h
ng
hp
load
%d VO2max/Kg %d
VE
%d
(W)
bo
ml/ph/Kg
bo
(lt)
bo

Nhm

Trc
TN
Sau
TN

SD
X

SD

150
20,00
190
20,00

95
7,00
109
9,73

33,3
5,20
43,7
3,53

65
7,50
80
8,21

91
4,50
100
4,50

Chc nng tim

mch
VO2/HR %d
ml/m
bo

52
8,50
69
7,66

13,5
0,35
14,2
0,41

79
7,50
94
7,50

200
Sau TN , 190
180
160
Trc TN, 150
140
120

Sau TN , 100

100
Trc TN, 91
80
60
Sau TN , 43.7
40
Trc TN, 33.3

Sau TN , 14.2

20
Trc TN, 13.5
0
Cng sut vn ng load (W)

Sc bn a kh VO2max/Kg (ml/ph/Kg)

Chc nng h hp VE(lt)

Chc nng tim mch VO2/HR (ml/m)

Hnh X. 3.6.1.a. Biu so snh test gng sc vn ng vin nam i in kinh

H Ni thng qua cc ch s chc nng trc v sau
thi gian dng vin tng lc Bionamine.
Qua kt qu bng trn cho ta thy nhng vn ng vin nam c dng vin
tng lc Bionamine trc v sau 1 thng luyn tp cc test kim tra u cho cc
kt qu r nt hn.

632

Bc cng sut VV thc hin c l c s chnh nh gi kh nng

gng sc. Test gng sc c thc hin trn xe p lc k theo bc cng sut tng
dn. Nam gii mi bc cng sut tng 50 W, thi gian mi bc cng sut ko di
2 pht. Kt qu cho thy cng sut vn ng thc t trc khi th nghim t
c l 150 W, t 95 % so vi d bo, sau vn ng thc t t 190 W t 109
% so vi d bo. Tng 14 %.
Sc bn a kh gia trc v sau tng 10.4 (ml/ph/Kg), chc nng h hp
kh nng d bo 17 %. Kh nng vn chuyn oxy ca h tim mch c nh
gi thng qua ch s oxy/mch (VO2/HR) l th tch oxy c tim bm vo mu
sau mi ln tim co bp. n v tnh l ml/mch p (ml/m), chc nng tim mch
gia trc v sau thc nghim tng 0.7 (ml/m). Ch s ny cho thy sau thc
nghim l rt tt.
T nhng kt qu trn cho thy sau 1 thng dng vin tng lc Bionamine
cc ch s test th lc ca vn ng vin nam c ci thin r rt.
Bng X.2.6.1.g. Cc ch s chc nng ca vn ng vin n i in kinh H Ni
trong test gng sc ti a.
Cng sut vn
Sc bn a kh
Chc nng h
ng
hp
Ch tiu
Load % d VO2max/Kg %d VE
%d
(W)
bo
ml/ph/Kg
bo
(lt)
bo
Nhm

Trc
TN
Sau
TN

150
86
28,0
SD 19,6
7,39
3,75
180
95
37,4
X
SD 20,0 10,63
5,20
Cc gi tr thng k vi n=14 v p0,05

67
8,11
87
7,50

180

44
4,41
75
4,50

Chc nng tim

mch
VO2/HR %d
ml/m
bo

30
6,23
60
8,50

11,2
0,42
15,0
0,35

125
7,35
107
7,50

Sau TN , 180

160
Trc TN, 150
140
120
100
Sau TN , 75

80
60
Sau TN , 37.4

40

Trc TN, 44

Trc TN, 28
Sau TN , 15
Trc TN, 11.2

20
0
Cng sut vn ng load (W)

Sc bn a kh VO2max/Kg
(ml/ph/Kg)

Chc nng h hp VE(lt)

Chc nng tim mch VO2/HR

(ml/m)

Hnh X.2.6.1.b. Biu so snh test gng sc vn ng vin n i in kinh H

ni thng qua cc ch s chc nng trc v sau thi gian dng
vin tng lc Bionamine.
Qua kt qu bng trn cho ta thy nhng vn ng vin n c dng b
sung vin tng lc Bionamine trong thi gian 1 thng tp luyn cc test kim tra
u cho cc kt qu r nt hn.

633

Bc cng sut VV thc hin c l c s chnh nh gi kh nng

gng sc. Test gng sc c thc hin trn xe p lc k theo bc cng sut tng
dn. n mi bc cng sut tng 30 W, thi gian mi bc cng sut ko di 2
pht. Kt qu cho thy cng sut vn ng thc t trc khi th nghim t c
l 150 W, t 86% so vi d bo, sau vn ng thc t t 180 W t 95% so vi
d bo. Tng 9%.
Sc bn a kh gia trc v sau tng 9,4 (ml/ph/Kg), chc nng h hp
tng gia trc v sau thc nghim l 31lit kh nng d bo tng sau thc nghim
l 60% tng 30%. Kh nng vn chuyn oxy ca h tim mch c nh gi
thng qua ch s oxy/mch (VO2/HR) l th tch oxy c tim bm vo mu sau
mi ln tim co bp. n v tnh l ml/mch p (ml/m), tng gia trc v sau
thc nghim l 3,8 (ml/m).
Cc kt qu trn chng minh rng sau 1 thng dng vin tng lc
Bionamine cc ch s test th lc ca vn ng vin n c ci thin r rt.
g. Tc dng tng cng th lc v nng cao thnh tch ca vn ng vin in
kinh ca vin tng lc Bionamine.
Thc nghim c chng minh trn 48 VV in kinh c chia 2 nhm
(C v TN), mi nhm li c chia theo nhm nam v n. Nhm TN dng vin
tng lc Bionamine, nhm C dung vin Placebo. Thi gian theo di l 30 ngy
thc nghim.
Kt qu trong bng X.2.6.1.h cho chng ta thy hu ht cc VV dng
vin tng lc Bionamine thnh tch u c ci thin r rt (C ly 100 m: VV
n tng trung bnh 0"37, VV nam tng trung bnh 0"38. C ly 200 m VV n
tng trung bnh 0"42, VV nam tng trung bnh 0"45. C ly 400 m VV n tng
trung bnh 102, VV nam tng trung bnh 143. Trong khi VV cc nhm
C hu nh khng c s thay i. iu vin tng lc Bionamine thc s c
tc dng tng cng b sung cc hot cht kch thch tng cng th lc, nng
cao thnh tch ca VV. Kt qu trn ph hp vi cc kt qu nghin cu ca cc
tc gi trc y [332, 337, 346, 349, 353].
Bng X.2.6.1.h. Kt qu kim tra thnh tch ca cc VV trc v sau khi dng
vin tng lc Bionamine. (n=14, p0.05)
Ch tiu

Nhm VV

VV TN
(N): X
VV
C(N): X
VV
TN(Nam): X
VV C
(Nam): X

C ly 100 m (s)
Trc Sau
SS
TN TN 30
ngy
13,29 12,92 0,37
0,10 0,10 0,01
13,27 13,26 0,01
0,04 0,04 0,00
11,84 11,46 0,38
0,14 0,14 0,00
11,88 11,84 0,03
0,04 0,03 0,01

C ly 200m (s)
Trc Sau
SS
TN TN 30
ngy
26,43 26,01 0,42
0,26 0,27 0,04
26,13 26,10 0,03
0,06 0,07 0,01
22,53 22,08 0,45
0,16 0,11 0,05
22,55 22,52 0,03
0,01 0,01 0,01

634

C ly 400m (s)
Trc Sau
SS
TN TN 30
ngy
57,92 56,9
1,02
0,24 0,24 0,01
56,98 56,94 0,04
0,12 0,13 0,01
51,18 49,75 1,43
0,27 0,25 0,08
50,54 50,50 0,04
0,12 0,12 0,01

X.2.6.2. nh gi v tc dng tng cng thn kinh phn x tr nh trn vn

ng vin
Bng X.2.6.2. Kt qu kim tra phn x ca VV bng bn - i hc TDTT T
Sn
Ch tiu

Phn x n
Ln 1
Ln 2
(ms)
(ms)

Ln 1
(ms)

MaX

227,84

176,20

352,97

276,00

5,00

3,00

Min

155,84

95,00

245,51

165,00

1,00

0,00

192,15

131,70

306,38

204,38

3,00

0,81

SD

21,34

25,14

29,25

28,08

1,17

0,81

MaX

256,18

227,69

421,00

219,84

6,00

3,00

Min

190,39

136,92

190,39

133,15

1,00

0,00

223,97

178,19

268,05

167,02

3,38

0,88

SD

21,26

26,00

59,25

21,52

1,17

0,78

Nhm

VV
nam

VV
n

Phn x phc
S li
Ln 2 (ms)
Ln 1

S li
Ln 2

Gi tr thng k sinh hc vi n=14, p0.05

* (ms): miligiy
* Ln 1: Kim tra trc t tp luyn.
* Ln 2: Kim tra sau 1 thng tp luyn

Kt qu kim tra phn x trong bng X.2.6.2 cho thy test kim tra phn
x n cc VV nam trc v sau thi gian tp luyn 1 thng trung bnh t
192,15 ms xung 131,70 ms gim 31%. Cn vn ng vin n gim trung bnh
t 223,97 ms xung 178,19 ms gim 20.44%.
Phn x phc ca vn ng vin nam sau 1 thng dng vin tng lc
Bionamine c ci thin trung bnh t 306,38 ms xung 204,38 ms gim
33,29%. Cn vn ng vin n phn x phc t 268,05 ms xung cn 167,02
ms gim 37,69%.
S li mc phi trong phn x phc ca VV nam c ci thin trung
bnh t 3 ln xung cn 0,81 ln. VV n gim trung bnh t 3,38 ln xung
cn 0,88 ln.
Cc kt qu kim tra v phn x n, phn x phc v s ln mc li in
t gia trc v sau khi dng h tr vin tng lc Bionamine c s khc bit nh
trn ta c th nh gi c tc dng ca vin tng lc Bionamine gip cho
cc vn ng vin tng hiu sut vn ng, kch thch hot ng ca thn kinh
trung ng x l tt phn x trong cc bi tp. Kt qu ny cng ph hp vi
nhiu tc gi [332, 334, 335].

635

Hnh X.2.6.2. Kim tra ch tiu phn x trn vn ng vin in kinh

X.2.7. tho lun
Mi qu trnh trao i cht u thng qua hot ng ca h thng enzym
v chu s phi hp iu ho ca h thn kinh, ni tit, v h iu tit t ng
trong t bo (iu khin cc phn ng ph hp vi yu cu ca c th). C th
con ngi l c mt h thng phc tp, c hng trm nghn cc phn ng ho hc
khc nhau. H thng ny hot ng rt chnh xc, ng b, nhanh nhy theo cc
tc ng mi trng bn ngoi v bn trong ca c th. cc phn ng trong c th
hot ng c lp hay lin kt v to thnh mt qu trnh. l nn tng ca s
sng..
Nhng tc ng lm bin i qu c cht v thnh phn ni mi trong c
th s c iu chnh bi tc ca cc phn ng ho sinh trong mt gii hn
nht nh. Trong iu kin thnh phn ni mi ca c th khi qu c cht b ph
hu v gim xung th mi s vn ng hay iu tit s t ng iu tit cn
bng. Vn ny c chng minh bng cc thc nghim trn vn ng vin
trong iu kin vn ng cng cao, khi thnh phn ni mi b ph hu v qu
c cht b gim xung sau 1 thng tp luyn v thi u. Hiu qu phc hi , thm
ch phc hi nhanh 1 s cc ch tiu v th lc cc vn ng vin di nh hng
tc dng ca vin tng lc Bionamine. iu ny th hin cc im di y:
- Cc hot cht sinh hc c trong thnh phn ca vin tng lc Bionamine
cung cp v b sung kp thi cho c th cc ngun hocmon Testosteron t
nhin, m khng phi l tng hp ho hc. Vai tr ca testosteron gip cho c
th tng thc y sinh tng hp protein, tng tch lu glucogen, y mnh chuyn
ho nit, photpho..v..v..
- Vin tng lc Bionamine cung cp cho c th sn phm giu cht vi
khong dng lin kt hu c (Fe, Zn, Cu) nhm tng to mu, thc y sinh
tng hp hemoglobin, feritin, cc enzym h hp v trao i cht.
- Vin tng lc Bionamine cung cp cho c th VV cc hot cht c
hot tnh sinh hc c tc dng tng hot ho enzym LDH gii phng axit lactic v
do c th nng cao c ngng chu ng axit lactic, tng nhanh qu trnh
636

hi phc ca c th. Vin tng lc Bionamine c tc dng tng cng cng

sut vn ng, chc nng tim mch v h hp gip cho vn ng vin tng cng
th lc tt hn.
X.3. Kt lun.
1. Cc kt qu nghin cu trn chng minh cho thy trong ngun nguyn liu
3 loi sao bin - Comb Sea Star, Spiny, Gallopping rt giu cc hot cht axit
amin, cc nguyn t vi lng v hot cht hoocmn steroit cn thit v c gi tr
b dng cho sc kho con ngi.
2. Vic ng dng cng ngh ho sinh enzim v s dng enzim papain thu
phn lm giu thm hm lng cc hot cht sinh hc c trong nguyn liu
loi sao bin - Sao bin gai lc (Comb Sea Star - Astropecten velilaris). lm
cho hm lng cc hot cht axit amin, cc nguyn t vi lng v hot cht
hoocmn steroit l cao nht. iu ny chng t cho thy sao bin Gai lc l
ngun nguyn liu qu cho ngnh ch bin thc phm chc nng v cng nghip
dc.
3. Sn phm bt nguyn liu Bionamine qua thc nghim trn ng vt cho
thy an ton v c tc dng y l thnh phn rt tt ca vin tng lc Bionamine,
l nhng ngun thc phm chc nng v dc liu c gi tr. Th hin cc hm
lng axit amin, hoocmn steroit v cc nguyn t vi lng l cao so vi mt s
loi sn phm tng t khc trong nc v nc ngoi.
4. Vin tng lc Bionamine an ton v c tc dng thc s r nt thng qua
vic thc nghim trn chut.
5. Vin tng lc Bionamine thc s c tc dng i vi vn ng vin in
kinh trong vic hon thnh tt cc bi tp th lc, kch thch phn x thn kinh
hot ng tt hn, tng cng c cc t cht v sinh l, sinh ho cho vn ng
vin v nng cao c thnh tch tp luyn ca cc vn ng vin.
6. Vic ng dng vin tng lc Bionamine trong vic tng cng th lc v
hi phc nhanh sc kho cho vn ng vin nhm nng cao thnh tch trong tp
luyn v thi u l rt ph hp v c hiu qu mang tnh mi v khoa hc v cng
ngh.
7. Vin tng lc Bionamine qua thi gian thc nghim cho thy s dng an
ton v yn tm trn vn ng vin, khng c phn ng ph xy ra. ngh B
Vn ho - Th thao v Du lch cn c trin khai dng rng ri cho vn ng
vin ti cc gii thi u trong nc v quc t nhm tng cng th lc v hi
phc sc kho nng cao thnh tch ca nn th thao nc nh.

637

Chng XI. Nghin cu bo ch v nh gi tc dng

bi b c th ca vin nang bo ng (Haliotis)
XI.1. m u
Nn y hc c truyn ca Vit Nam c mt truyn thng v c lch s lu
i phong ph. T xa ng cha ta bit s dng ngun dc qu gi ca t
nc vi cc phng php ch bin khc nhau, vi cc dng bo ch thch hp
dng phng v cha bnh cho nhn dn, trong vic khai thc s dng
ngun dc liu t sinh vt bin chim mt t l ng k.
Ngy nay, cng vi s pht trin ca khoa hc k thut, m hnh nghin
cu lin ngnh gia cc nh khoa hc thuc cc lnh vc ho sinh v Y Dc
nhm khai thc v chuyn dng bo ch thch hp t ngun dc liu bin
c s dng c hiu qu nhiu nc trn th gii nh Nht Bn, Hn Quc,
Trung Quc, Niu Di Ln, M, c rt nhiu thuc c ngun gc t sinh vt
bin c mt trn th trng dng iu tr bnh, bi b c th, b sung axit
amin v cc nguyn t vi lng do cc hng sn xut dc phm trn th gii
cung cp nh thuc h tr iu tr ung th Cytarabin (Ara-C) c chit tch t
loi hi min Cytotethy cryta, thuc khng sinh Phycoerythin c ngun gc t to
(Red algae), ngoi ra cn tin hnh chit xut v phn lp, thu phn t rt
nhiu ngun dc liu t sinh vt bin vi tr lng ln c kh nng khai thc v
nui trng nh rong bin, hi sm, bo ng, c nga Nhng thnh qu ny
mang li li ch kinh t rt to ln cho nhiu Quc gia trn th gii.
Vit Nam c b bin tip gip hu ht chiu di t nc, vi hn mt
triu km2 vng bin, c kh hu nhit i gi ma, l iu kin tt cho cc dng
sinh vt bin sinh sng, pht trin v thun li cho vic nui trng trong c
bo ng. T nhng nm 1970 Vit Nam c mt s cng trnh nghin cu
chuyn dng bo ch t nguyn liu l sinh vt bin nhng cn t i v hn ch.
Bo ng cn gi l cu khng hay c khng c tn khoa hc l Haliotis
thuc h Halitidae, l loi c c v cng, hi dt, mt ngoi nhm v c mu
nu sm, mt trong c lp x c ng nh. mp c 7 13 l. y l mt loi hi
sn qu, n l mt trong 8 mn n cao cp gi l bt trn thng hin din
trong nhng ba tic cung nh hay nhng gia nh giu c.
Vi mc ch khai thc v sng lc sinh vt bin c hot tnh sinh hc
nhm bo ch dng thuc t bo ng thch hp, tin s dng v c hiu qu kinh
t. Vin ha hc cc hp cht thin nhin Vin khoa hc Vit Nam phi hp
vi Vin Y hc c truyn Qun i B Quc phng tin hnh: Nghin cu bo
ch v nh gi tc dng bi b c th ca vin nang Haliotis t bo ng vi
mc ch:
1. Nghin cu qui trnh bo ch vin nang mm HALIOTIS t bo ng
2. Nghin cu c tnh v nh gi tc dng bi b c th ca vin nang
HALIOTIS.

638

XI.2. kt qu v tho lun

XI.2.1. Kt qu nghin cu qui trnh bo ch vin nang
HALIOTIS t Bo ng
Bo ng

X l sch tp cht

Sy kh (50-60o)

Nghin bt mn

Trn t dc

ng nang mm

Sy nang

ng l, p v, hon thin
sn phm
Kim nghim
thnh phm
Nghin cu
Hnh XI.2.1.a. S quy trnh sn xut vin nang mm bo ng

639

- Nghin cu la chn dc liu v xy dng qui trnh bo ch vin nang mm

Bo ng Haliotis thnh dng bn thnh phm v thnh phm: xy dng
c qui trnh bo ch sn phm bo ng di dng nang mm nh sau:
Cng thc cho 1 vin:
Bt kh bo ng: 150 mg
T dc v:
01 vin
Tin hnh sn xut th nghim c 67.000 vin nang trn dy chuyn sn
xut nang mm hin i lm sn phm mu v phc v cho cng tc p dng trn
lm sng v ng k lu hnh sn phm.
- Nghin cu xy dng tiu chun c s cho sn phm vin nang mm bo
ng Haliotis: tin hnh nghin cu xy dng tiu chun c s ph hp vi ch
tiu do B Yt qui nh cho sn phm.
+ Cn c quy ch ghi nhn hng ho lu thng trong nc v hng ho xut
khu, nhp khu ca th tng chnh ph
+ Cn c quy ch cng b tiu chun sn phm thc phm ban hnh km theo
quyt nh s 42/2005/Q-BYT ngy 08 Thng 12 nm 2005 ca B trng B
Y t
Chng ti tin hnh xy dng tiu chun c s p dng cho sn phm thc
phm chc nng vin nang mm bo ng Haliotis vi cc ch tiu nh sau:
a. Cc ch tiu cm quan.
- Trng thi : Dng vin nang mm
- Mu sc : c trng cho sn phm
- Mi v : Thm c trng cho sn phm
b. Cc ch tiu l ho:
STT
Tn ch tiu
n v tnh
Mc cng b
1
Hm lng m
%
<3
2
Khi lng vin
mg
1600-1800
3
Hm lng carbohydrat
%
<1
4
Hm lng Protein
%
22-27
5
Hm lng Lipit
%
52-57
6
Hm lng tro ton phn
%
<1
c. Cc ch tiu v sinh:
TT
Tn ch tiu
1
Tng s vi sinh vt hiu kh
2
E. coli
3
Coliforms(MPN/ml)
4
Cl. perfringens
5
B. cereus
6
S. aureus
7
Salmonella
8
TSBTNM-M

n v tnh
vi khun/ g
vi khun / g
vi khun /g
vi khun / g
vi khun / g
vi khun / g
vi khun /25g
Bo t/g

640

Mc ti a
104
0
10
0
0
0
0
102

d. Hm lng kim loi nng:

TT
Tn ch tiu
1
Asen (As)
2
Ch (Pb)
3
ng (Cu)
4
Thic (Sn)
5
Km (Zn)
6
Thu ngn (Hg)
7
Cadimi (Cd)
8
Antimon (Sb)

n v tnh
mg/ kg
mg/ kg
mg/ kg
mg/ kg
mg/ kg
mg/ kg
mg/ kg
mg/ kg

Mc ti a
0,1
0,5
5
40
40
0,05
1
1

e. Hm lng cc cht c hi khng mong mun:

TT
Tn ch tiu
n v tnh

Mc cng b

1
2
3

c t Aflatoxin B1
ppb
< 10
c t vi nm khc
ppb
< 35
Ho cht bo v thc vt ph hp vi quy nh ti Quyt nh s
867/1998/Q-BYT ngy 04/4/1998 ca B trng B Y t

f. Thnh phn cu to:

Bt kh nguyn cht Bo ng, t dc, Gelatine dng lm v nang.
g. Thi hn s dng:
24 thng k t ngy sn xut. Ngy sn xut v hn s dng ghi trn
nhn sn phm.
h. Hng dn s dng Bo qun
Cng dng: Bi b sc kho, tng cng sinh lc, gip tng th lc, sng
mt v h tr iu tr u chong, au nhc xng khp.
Ch : Thc phm ny khng phi l thuc, khng c tc dng thay th
thuc cha bnh.
Liu dng: Ngy dng 2 vin/ln x 2 3 ln /ngy
Hng dn bo qun, vn chuyn: ni kh ro, thong mt, trnh nh
sng mt tri.
i. Cht liu bao b v quy cch bao gi:
Cht liu bao b:
ng l PE chuyn dng, p v m bo yu cu v sinh thc phm.
Qui cch ng gi:
Khi lng tnh:
+170 gam/l (1700mg/vin x 100 vin)
+ 17gam/v (1700mg/vin x 10 vin x10 v/hp)
Xut x: Sn xut bi Vin Y hc c truyn Qun i
j. D tho ni dung ghi nhn sn phm
Tn sn phm: Thc phm chc nng: Vin nang mm bo ng
Thnh phn cu to:
Bt kh nguyn cht Bo ng, t dc, Gelatine dng lm v nang

641

Ch tiu cht lng ch yu:

Hm lng Protein
%
22 - 27
Hm lng carbohydrat
%
<1
Hm lng cht bo
%
52 - 57
Ngy sn xut, hn s dng: Xem trn bao b
Cng dng:
Bi b sc kho, tng cng sinh lc, gip tng th lc, sng mt v h tr
iu tr u chong, au nhc xng khp.
Ch : Thc phm ny khng phi l thuc, khng c tc dng thay th thuc
cha bnh.
Liu dng: Dng 2 vin/ln x 2 3 ln /ngy
Hng dn bo qun, vn chuyn: ni kh ro, thong mt, trnh nh sng
mt tri.
Khi lng tnh:
+ 170 gam/l (1700mg/vin x 100 vin)
+ 17gam/v (1700mg/vin x 10 vin x10 v/hp)
- Tin hnh cng b sn phm di dng thc phm chc nng vi B y t
xin cp s ng k lu hnh ton Quc:
Sn phm c Cc An ton v sinh thc phm B Y t cp chng nhn
tiu chun sn phm cho vin nang mm bo ng s: 6714/2007/YT-CNTC. Ph
hp vi cc qui nh hin hnh v cht lng, v sinh an ton thc phm v c
php lu hnh ton quc ng nh tiu chun cng b.

Hnh XI.2.1.b. Chng nhn tiu chun sn phm vin nang mm bo ng

642

Hnh XI.2.1.b. Sn phm vin nang mm bo ng

XI.2.2. kt qu Nghin cu c tnh ca vin nang Haliotis
XI.2.2.1. Th c tnh cp :
Bng XI.2.2.1. B tr th nghim v kt qu th c tnh cp:
Nhm
1
2
3
4
5
6

Liu dng
(ml/ 10g chut)
0,2 ml x 1 ln
0,2 ml x 3 ln
0,3 ml x 3 ln
0,4 ml x 3 ln
0,5 ml x 3 ln
0,5 ml nc x 3 ln

Liu dng
(g / kg chut)
2,0 g
6,0 g
9,0 g
12,0 g
15,0 g
nc ct

S chut th
nghim
10
10
10
10
10
10

S chut
cht
0
0
0
0
0
0

Nhn xt: Sau khi ung nc ct v mu th, chut tt c cc nhm u

hot ng, n ung bnh thng k c nhm ung 3 ln vi th tch ti a cho
chut th nghim (0,5 ml/ 10g chut/ln). Khng c chut no cht trong thi
gian theo di.
XI.2.2.2. Th c tnh bn trng din
Tnh trng th: Trong thi gian th nghim, hu ht th c hai nhm hot ng
bnh thng, n ung tt, phn kh, lng mt; khng c hin tng rng lng
hoc kh cng.
Quan st i th cc t chc: Khi kt thc th nghim, tin hnh m th, quan st
cc t chc gan, thn, tim, phi. Khng nhn thy biu hin khc thng trn cc
t chc ca th nhm th so vi nhm chng.

643

Quan st m bnh hc trn tiu bn vi th: Phn tch m bnh hc c thc hin
ti Khoa Nghin cu thc nghim - Vin Y hc c truyn Qun i
Hnh thi vi th gan sau ung vin nang Haliotis:
- L chng: cu trc ca gan bnh thng.
- L th nghim: cc b gan, t bo khng hoi t. Cc xoang mch khng
gin, khng xung huyt.
Hnh thi vi th thn:
- L chng: Cc cu thn khng xung huyt
- L th nghim: cu thn v ng thn bnh thng, khng thy thoi ho biu
m ng thn
Kt qu cn nng th v cc ch s huyt hc, ho sinh c ghi trn cc
bng XI.2.2.2.a-c.
Bng XI.2.2.2.a. Kt qu theo di cn nng th
Nhm

Chng
Th
P (chng-th)

10
10

Cn nng (kg)
Trc th nghim
Sau th nghm

2,16 0,17
2,18 0,14
0,162

2,19 0,15
2,21 0,18
0,120

P
(trc - sau)

0,036
0,024

Bng XI.2.2.2.b. Kt qu theo di cc ch s huyt hc

Ch tiu nh gi

Hng cu
(x 1012/ l)
Bch cu
(x 109/ l)
Huyt sc t
(g/l)
Hematocrit
(%)

Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)

Trc
th nghim

Kt qu
Sau
th nghim

10
10

5,90 0,20
5,88 0,23
0,389
6,22 0,26
6,17 0,20
0,069
127,1 2,6
124,1 3,5
0,201
38,1 1,9
38,5 2,4
0,306

5,92 0,28
5,95 0,21
0,313
6,19 0,21
6,24 0,18
0,095
122,0 5,1
118,8 4,5
0,213
37,1 2,1
37,6 2,2
0,212

10
10
10
10
10
10

P
(trc- sau)

0,494
0,005
0,363
0,002
0,058
0,023
0,018
0,056

Bng XI.2.2.2.c. Kt qu theo di cc ch s sinh ho

Ch tiu nh gi

Protein
ton phn
(g/ l)
Bilirubin

Chng
10
Th
10
P (chng-th)
Chng
10

Trc
th nghim

Kt qu
Sau
th nghim

49,6 4,5
54,9 5,1
0,232
3,68 0,39

53,5 3,6
49.8 4,2
0,187
3,36 0,52

644

P
(trc - sau)

0,309
0,257
0,032

(mol/ l)
GOT
(UI/l)
GPT
(UI/l)
Ure
(mmol/l)
Creatinin
(mol/ l)

Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)

10
10
10
10
10
10
10
10
10

3,72 0,27
0,457
50,5 4,5
38,4 6,1
0,048
58,5 10,7
62,7 11,4
0,467
5,38 1,32
4,67 1,21
0,143
105,2 11,3
107,6 9,8
0,361

2,89 0,34
0,239
58,6 5,7
49,5 4,9
0,105
67,6 10,8
80,3 12,2
0,121
5,28 1,03
4,42 0,72
0,122
109,8 12,1
113,2 7,5
0,206

0,012
0,113
0,103
0,207
0,019
0,502
0,426
0,181
0,059

XI.2.3. nh gi tc dng bi b c th ca vin nang bo

ng
XI.2.3.1. Ni dung nghin cu
- Nghin cu tc dng bi b sc kho, tng cng sinh lc, gip tng th lc,
sng mt v h tr iu tr u chong, au nhc xng khp ca vin nang Haliotis
trn lm sng.
- Theo di tc dng khng mong mun ca vin nang mm Haliotis.
XI.2.3.2. nh gi hiu qu h tr iu tr chng h lao sau 4 tun
a. S ci thin cc triu chng lm sng trc v sau thc nghim
au u, hoa mt, chng mt, tai , mt ng, hay qun, th trng gy; ngi
mt mi, hay cu gt, lng bn tay chn nng; ra m hi trm; au lng; mi gi;
nhc trong xng.
Bng XI.2.3.2.a. S ci thin ca cc triu chng trc v sau thc nghim
Thi gian
Triu chng lm sng

Mt mi
au u
Chng mt, hoa mt
Hay qun
Ng tm phin nhit
au lng, mi gi

Do

D30

S lng BN

T l

S lng BN

T l

30
21
18
16
9
14

30/45
21/45
18/45
16/45
9/45
14/45

8
10
9
7
3
4

8/45
10/45
9/45
7/45
3/45
4/45

* Nhn xt: Sau 4 tun thc nghim ta thy cc triu chng lm sng nh mt
mi gim t 30 cn 8 bnh nhn, au u gim t 21 cn 10 bnh nhn chng
mt, hoa mt gim t 18 cn 9 bnh nhn, hay qun gim t 16 cn 7 bnh nhn,
ng tm phin nhit gim t 9 cn 3 bnh nhn, au lng mi gi gim t 14 cn
4 bnh nhn.

645

b. S thay i im trung bnh Crocq - Bugard trc v sau thc nghim

Bng XI.2.3.2.b. S im trung bnh Crocq - Bugard sau 4 tun thc nghim (n =
45)
Thi im nghin cu

S im Crocq - Bugard

D0

D30

39,73 1,91

26,21 1,11
34%
p0-30< 0,001

( x s)
% gim
p

S im trung bnh Crocp - Bugard trc thc nghim l 39,73, sau 4 tun
thc nghim l 26,21 (gim 34%); s im gim c ngha thng k vi p <
0,001.
c. S thay i im trung bnh Wechsler trc v sau thc nghim
Bng XI.2.3.2.c. S im trung bnh Wechsler sau 4 tun thc nghim (n = 45)
S im Wechsler

Thi im nghin cu
D30

D0

( x s)
% tng
p

48,49 1,16

63,16 1,28
30%
p0-30 < 0,001

S im trung bnh Wechsler trc thc nghim l 48,49, sau 4 tun thc
nghim l 63,16 (tng 30%); s im tng c ngha thng k vi p < 0,001.
d. S thay i cn nng trung bnh trc v sau thc nghim
Bng XI.2.3.2.d. S thay i cn nng trung bnh sau 4 tun thc nghim (n = 45)
Cn nng
(kg)

D0

Thi im nghin cu
D30

( x s)

42,96 6,79

43,35 6,64
p0-30 < 0,05

Cn nng trung bnh trc thc nghim l 42,95 kg, sau 4 tun thc
nghim l 43,35 kg, s thay i cn nng trung bnh c ngha thng k vi p <
0,05.
e. S thay i c lc trung bnh trc v sau thc nghim
Bng XI.2.3.2.e. S thay i c lc tay phi trung bnh sau 4 tun thc nghim (n
= 45)
C lc tay phi
(kg)
( x s)

Thi im nghin cu
D0

D30

18,45 5,47

19,75 5,18
7%
p0-30 < 0,05

%tng
p

C lc trung bnh tay phi trc thc nghim l 18,45 kg,sau4 tun iu l
19,75 kg (tng 7%); s tng c lc trung bnh c ngha thng k vi p < 0,05.

646

Bng XI.2.3.2.f. S thay i c lc tay tri trung bnh sau 4 tun thc nghim (n =
45)
C lc tay tri
(kg)
( x s)

Thi im nghin cu
D0

D30

17,14 5,67

18,42 5,75
% tng
7,5%
p
p0-30 < 0,05
C lc trung bnh tay tri trc thc nghim l 17,14 kg, sau 4 tun thc
nghim l 18,35 kg(tng 7,5%); s tng c lc trung bnh c ngha thng k
vi p < 0,05.
f. S ci thin v n trc v sau thc nghim
Bng XI.2.3.2.g. S ci thin v n sau 4 tun thc nghim (n = 45)
Thi im nghin cu

S ci thin
v n

D0

n
2
18
25

Tt
Trung bnh
Km

p0 30
D30

%
4%
40%
55%

n
18
17
10

%
40%
37%
22%

< 0,05

Sau 4 tun thc nghim s bnh nhn n tt tng t 2 bnh nhn ln 18, c
ngha thng k vi p < 0,05, s bnh nhn n km gim t 25 bnh nhn xung
cn 10, c ngha thng k vi p < 0,05.
g. S ci thin v ng trc v sau thc nghim theo thang im Pittsburgh
Bng XI.2.3.2.h. S ci thin v ng sau 4 tun thc nghim (n = 45)
S ci thin
v ng

Tt
Trung bnh
Km

D0

n
2
22
21

p0 30

Thi im nghin cu
D30

%
4%
48%
46%

n
18
22
5

%
40%
48%
11%

< 0,05

Sau 4 tun thc nghim s bnh nhn ng tt tng t 2 bnh nhn ln 18,
c ngha thng k vi p < 0,05, s bnh nhn ng km gim t 21 bnh nhn
xung cn 5, c ngha thng k vi p < 0,05.

647

XI.2.3.3. Tc dng ca vin nang Haliotis trn chc nng gan, thn, nhm
m mu v huyt hc
a. nh gi tc dng ca vin nang Haliotis trn chc nng gan, thn, nhm
m mu
Bng XI.2.3.3.a. So snh phn tch ch s m mu trc v sau thc nghim
( x s)
Nhm

n=45

Trc T
Sau T

TC

TG

HDL-C

LDL-C

4,760,21
3,810,34
<0,05

2,130,26
1,650,04
<0,05

0,920,12
1,330,22
<0,05

2,650,13
2,120,35
<0,05

Qua kt qu bng XI.2.3.3.a ta c th thy sau 4 tun thc nghim thuc

c tc dng lm lng TC, TG, LDL-C gim v lm tng lng HDL-C trong
mu, c ngha thng k vi p < 0,05.
Bng XI.2.3.3.b. So snh phn tch tnh hnh chc nng gan trc v sau thc
nghim ( x s)
Nhm

n=45

Trc T
Sau T

AST

ALT

GGT

42,464,97
31,242,23
>0,05

28,542,73
27,131,34
>0,05

39,764,67
34,681,65
>0,05

Bng XI.2.3.3.c. So snh phn tch tnh hnh chc nng thn trc v sau thc
nghim ( x s)
Nhm

n=45

Trc T
Sau T

URE

CREA

5,091,07
5,160,68
>0,05

101,758,42
87,522,51
>0,05

Qua bng XI.2.3.3.b-c so snh gi tr trung bnh ca chc nng gan v thn
trc v sau thc nghim cho thy khng c thay i, khng c ngha thng k (p
> 0,05).
b. S thay i cc ch s huyt hc trc v sau thc nghim
Bng XI.2.3.3.d. S thay i cc ch s huyt hc trc v sau thc nghim ( x s)
Thi gian

S lng hng cu (M/ul)

Hemoglobin (g/dl)
S lng bch cu (K/ul)
Protit mu ton phn (g/l)

D0 ( x s)
4,11 0,12
12,34 0,32
7,40 0,45
69,271,51

D60 ( x s)
4,62 0,05
13,130 0,28
8,05 0,37
71,84 1,36

< 0,001
< 0,05
> 0,05
< 0,05

Sau 4 tun thc nghim nhn thy s tng hng cu trong mu ngoi vi c
ngha thng k (p < 0,001), tng Hb trong mu ngoi vi c ngha thng k (p
< 0,05), tng protit mu c ngha thng k (p < 0,05). So snh gi tr trung
bnh trc v sau thc nghim thy s thay i bch cu khng c ngha
thng k (p > 0,05).
648

XI.2.3.4. Theo di tc dng khng mong mun ca vin nang mm Bo ng

trn lm sng:
Trong qu trnh thc nghim tt c cc bnh nhn u khng thy xut
hin cc triu chng bun nn, a chy, y bng, mn nga.
XI.2.3.5. Nhn xt v bn lun
Sau 4 tun thc nghim ta c th d nhn thy cc triu chng lm sng
c ci thin r rt: Mt mi gim t 30 cn 8 bnh nhn, au u gim t 21
cn 10 bnh nhn chng mt, hoa mt gim t 18 cn 9 bnh nhn, hay qun
gim t 16 cn 7 bnh nhn, ng tm phin nhit gim t 9 cn 3 bnh nhn, au
lng mi gi gim t 14 cn 4 bnh nhn (bng XI.2.3.2.a).
Qua kt qu t bng XI.2.3.2.b-h ta thy mc suy nhc, ri lon tr
nh, n v ng c ci thin r rt. Th trng, trng lc c u tng sau 4 tun
thc nghim.
Bng XI.2.3.3.b-c th hin khng c s bin i chc nng gan v thn.
Ngoi ra qua qu trnh thc nghim thy lng m trong mu gim, tng s
lng hng cu, hemoglobin v protit mu ton phn. Khng thy xut hin cc
triu chng bun nn, a chy, y bng, mn nga.
XI.3. Kt lun
Vin nang Haliotis bo ch t Bo ng: c cng b lu hnh sn phm
di dng thc phm chc nng, c B Y t cp s ng k lu hnh ton
Quc. Qua nghin cu c tnh v nh gi tc dng trn lm sng cho thy:
1. V c tnh:
- Khng xc nh c liu gy cht 50% ng vt th nghim (LD50) v khng
tm c liu gy cht chut mc d cho ung vi mc liu cao.
- Cho th ung lin tc trong mt thng vi mc liu 2 vin/kg/ngy khng
nhn thy s khc nhau ng k v tnh trng th, cc ch s huyt hc v sinh
ho, i th cc tng ph sau th nghim gia nhm chng v nhm th; hnh nh
tn thng m bnh hc t chc gan v thn trn tiu bn vi th khng c s khc
nhau nhiu gia nhm chng v nhm th.
2. V tc dng bi b c th trn lm sng:
nh gi tc dng h tr iu tr bnh suy nhc c th ca vin nang mm
bo ng trn lm sng sau 4 tun thc nghim ta thy vin nang mm bo ng c
tc dng: tng cng sinh lc, gip tng th lc, sng mt v h tr iu tr u
chong, au nhc xng khp.

649

Chng XII. Nghin cu cng ngh sn xut vin

nang mm hi sm Hasamin
XII.1. M u
XII.1.1. Hi sm trong kinh nghim dn gian
Hi sm, hay cn c gi l da bin, sm bin, a bin, l loi ng vt
khng xng sng. Thn c dng ng di nh qu da, phnh ra gia v thon
nh hai u vi nhng gai tht nh. Pha trc c ming vi vnh tua r rt, pha
sau l hu mn. Dc thn l cc dy chn ng. Da mm c cc phin xng nm
ri rc di da. Hi sm l ng vt phn tnh, tr mt s loi thuc b Khng
chn (Apoda). Trng th tinh v pht trin ngoi c th m. Sng b trn nn y,
su khc nhau. Ch c mt s loi thuc h Pelagothuridae l bi li. Hi
sm c kh nng ti sinh rt cao v sc chu ng bn b (c th sng di nc
ti su 6.000m).
C nhiu loi hi sm thuc cc chi Holothuria, Actinopyga, Stichopus.
Trong , hai loi hi sm en v trng c s dng rng ri hn:
- Hi sm en (Holothuria vagabunda) c thn mu en, bng nht mu
hn, di 30-40cm.
- Hi sm trng (H. scabra) c lng mu xm, nht dn hai bn, bng
trng, di 40-50cm, c khi n 60-70cm.
Trn th gii, hi sm c hng nghn loi, phn b nhiu nc nh Trung
Quc, Nht Bn, c, n , Malaysia, vng ng Phi.... Vit Nam, c bn loi
c pht hin ph bin l hi sm en, hi sm trng, hi sm v (Microthele
nobilis Selenka) v hi sm mt (Actinopyga echinites Jaeger). Chng c khp
tuyn bin t bc vo nam, nhiu nht Ph Yn, Khnh Ha, Vng Tu, Cn
o, Ph Quc, Kin Giang. Chng thng sng cc di nng ven b n
su khong 5-7m, ch yu vnh v nhng ni nhiu ngm. Di chuyn bng
cch pht nc hoc co dui cc c. n ng vt, thc vt nh y v cht b
hu c.
Nhiu quc gia trn th gii t chc nui hi sm bo v v ch
ng ngun nguyn liu tiu dng.
Ton con hi sm bt v c m b rut, ra sch ri phi hay sy kh.
Loi tht c mu en qunh, dnh, da c nhiu gai bu l tt.
Theo cc sch thuc [354]: Tht hi sm c 21,45% protein, 0,27% lipit,
1,37% gluxit v 1,13% tro, trong tro ch yu gm canxi: 0,118; photpho: 0,22;
st: 0,0014; kali: 0,070.
Theo y hc c truyn, hi sm v mn, tnh m, c cng dng b thn
trng dng, t m dng huyt, ch tinh nhun to, thng dng cho nhng
ngi b suy nhc, lao lc, thiu mu, ph n sau khi sinh, thn dng h nhc
gy nn tnh trng lit dng, di tinh, xut tinh sm, di niu...
V mt thc phm, hi sm l loi bin c gi tr dinh dng rt cao,
rt giu cht m, cc axit amin, vitamin v cc nguyn t vi lng, c bit l
st v iode. Hm lng cholesterol trong hi sm rt thp, hu nh khng c, nn
hi sm l thc phm b dng rt tt cho nhng ngi b cao huyt p, ri lon

650

lipit mu, x va ng mch, ung th hoc c bnh l mch vnh... Sau khi ch
bin c mi v thm ngon, rt hp dn v thng c mt trong cc yn tic sang
trng nht Trung Quc, Nht Bn, n , Maylaysia, Philipin. Tht hi sm c
th n ti di dng xo nu, p mui, ng hp; nhng ch bin kh l
phng php ph bin nht. Hi sm phi kh cn l mt hng xut khu c gi
tr cnh tranh c vi yn so, bo ng.
V mt dc liu, do rt giu dinh dng v tc dng khng km v nhn
sm, nn ngi ta coi tht hi sm l nhn sm ca bin. N cha tnh trng c
th gy yu, thn kinh suy nhc, vim ph qun, cc chng chy mu, ho, mn
nht. Dng dng thng thng l nng gin, tn thnh bt. Ngy ung ba ln,
mi ln 6-10g vi nc m hay ru. Trung Quc, ngi ta dng hi sm di
dng thuc n thun hoc ch bin thnh thc n, v thuc nh sau:
- Cha lao phi: Hi sm (500g), bch cp (250g), mai ra (1 ci, nng
gin). Tt c thi nh, sao vng, tn bt, ry mn. Ngy ung 3 ln, mi ln 25g
vi nc m.
- Cha c th nhit to, khi u: Hi sm (30g), mc nh (30g), thi nh v
nhi vo mt khc rut gi ln, luc cho chn nh, n trong ngy.
- Thuc b kh huyt, h huyt p: Hi sm (50g, thi ming), ti (30g, gi
nh), go (100g, vo sch). Tt c nu k, nh thnh cho. n lm mt ln trong
ngy vo bui sng.
- Thuc b gan, thn, h huyt p: Hi sm: (50g), trng (5g) cho vo
ni cng vi nc dng g (100ml), t hnh, gng, mui. Nu cho nh tht. n
lm mt ln trong ngy.
nc ta, hi sm cng c ch bin thnh nhiu mn n bi thuc c
gi tr cao nh:
- Cho g Hi sm: Hi sm (kh 20g, ti 40g), tht g 100g, go 100g,
hnh hoa 10g, mui, gia v. Cch ch: Hi sm kh ngm trong nc m, ra
sch, ct ming nh, Hi sm ti ra sch, m b ni tng ct ming nh, tht g
ct thnh ming cho cng go i sch vo ni vi nc 1.000ml, dng la to
un si, sau un nh la nu thnh cho, cho hnh thi nh, mui, gia v va
n. Tc dng: n b t thn, ch kh dng huyt, dng cho trng hp thn h,
di tinh, mng tinh, to bn. Ngy n 2 bt chia 2 ln.
- Hi sm xo mp ng: Hi sm kh 100g (ti 200g), mp ng
500g, hnh hoa 50g, du n, mui, gia v va . Cch ch: Hi sm kh ngm
trong nc m, ra sch, ct ming nh, Hi sm ti ra sch, m b ni tng
ct ming nh, mp ng b rut thi nh, bp qua t mui. Cho du n v Hi
sm vo cho xo chn sau cho mp ng, mui, gia v va ; mn n c
mi v thm ca Hi sm v hnh. Cng dng: Hi sm v mp ng u i vo
thn kinh v can kinh, gip cho c th cng trng, b thn, trng dng, ch
tinh. Mn ny cn dng cho ngi b bnh i tho ng, mp ng v Hi
sm u b chn m gip cho ngi bnh i tho ng thy mt m trong
ngi, gim i, gim kht.

651

XII.1.2. Ngun nguyn liu hi sm

Theo Thng tn x Vit Nam, hi sm trong thin nhin ngy cng b cn
kit do nh bt qu mc, nhng hin loi c mnh danh l "nhn sm ca
bin" ny khng nhng ang c gy nui d dng m cn cho nng sut cao
ngoi s trng i ca ngi nui.
Khu bo tn bin Rn Tro, x Vn Hng, huyn Vn Ninh, tnh Khnh
Ha c xy dng vi s tr gip ca IMA VN (Lin minh Sinh vt Bin Quc
t ti Vit Nam) l mt m hnh nui hi sm rt thnh cng. Hi sm nui ti
y pht trin rt tt, khng b bnh, trng lng tng nhanh.
Qua thc t cho thy, nui hi sm khng tn thc n, khng mt cng
cho n v loi ny ch n mn b hu c, thc vt ph du. Loi ny c kh nng
tn ti cao, t dch bnh, c th kch thch sinh d dng v li ln nhanh.
Khng ch c vy, hi sm cn c kh nng ci to mi trng bin, lm sch
nc nn c th nui chung vi cc loi thy sn khc nh tm s.
T chc ICLARM (Trung tm Ngh c Th gii) cng h tr vn cho
Trung tm Nghin cu Thu sn III thc hin cng trnh nghin cu "Sn xut
nhn to v nui hi sm thng phm". Cng trnh chn hi sm ct (H.
scabra) v hi sm mt (A. echinites) nui, bc u mang li thnh cng ng
k.
Bn cnh , nhiu m hnh nui hi sm ca ngi dn cng thu c
nhng kt qu kh quan. Ti phng Cam Ngha, th x Cam Ranh, tnh Khnh
Ho mt s gia nh do nui tm s khng thnh cng, chuyn sang nui hi
sm. Ch sau 4 thng, hi sm c thu hoch v bn ra th trng vi gi hn
20.000 ng/kg tht ti, tr chi ph vn li gn 3 triu ng. Vn l mt loi thc
n c sn ngon v b, c th dng ch bin mn n cng nh cng nghip
dc liu nn hi sm c tiu th mnh c th trng trong ln ngoi nc.
XII.1.3. Nghin cu thnh phn sinh ha mt s loi hi sm
Trc y, trong khun kh ti cp nh nc KC09.15 (2003-2005),
chng ti tin hnh cc nghin cu xc nh thnh phn sinh ha ca cc loi
hi sm: hi sm en: Holothuria vagabunda, hi sm trng: H. scabra, hi sm
: Holothuria sp., hi sm nu: Actynopyga echinites, hi sm xanh:
Bohadschia graeffei, hi sm vng, hi sm da trn: Bohadschia argus (Jaeger)
v hi sm v: Microthele nobilis v thu c cc kt qu sau:

652

a) Hm lng protein tng v thnh phn axt amin (xc nh protein tng s theo
phng php Kjeldalh v Lowry, phn tch thnh phn axt amin bng phng php in di sc
k giy 2 chiu)

Mu nghin cu
STT
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17

Protein
Axt amin

H,
Actynopyga Holothuria Bohadschia Bohadschia
graeffei
argus
scabra
echinites
sp,
67,82
65,63
67,82
63,23
60,35
(%mu
(%mu
(%mu
(%mu
(%mu kh)
kh)
kh)
kh)
kh)

Aspactic axit
Glutamic axit
Xerin
Histidine
Glyxin
Threonine
Alanin
Agrinine
Tyrosine
Xistein+
Xistin
Valin
Metionin
PhenynAlanin
Izolxin
Lxin
Lizin
Prolin

5,42
10,25
2,35
2,48
3,38
5,74
2,92

4,62
9,25
1,80
2,08
3,15
4,20
2,35

5,30
9,66
2,20
2,40
3,80
5,50
3,05

5,58
7,85
1,84
2,12
2,56
5,96
2,34

4,80
6,40
1,84
0,07
8,62
2,29
3,75
3,22
1,14

2,82

2,78

2,70

3,82

4,57

2,78
2,02
6,88
8,78
8,66

2,02
3,82
7,74
9,78
8,02

2,58
3,44
8,84
9,24
8,06

2,72
2,46
6,25
8,48
7,68

1,52
0,57
0,89
0,97
1,64
0,68
2,71

b) Hm lng lipit v thnh phn axt bo (Xc nh Lipit v axt bo theo Tiu chun
ISO/DIS 659:1988 v ISO/FDIS 5590:1998, LB c. Phn tch trn my sc k kh HP-6890,
ghp ni vi Mass Selective Detector Agilent 5973.)

Hm lng lipit
STT
Tn Vit Nam
1
Hi Sm vng

Tn khoa hc
ang xc nh tn

Hm lng Lipit [%]

0,27a

Hi Sm v

Microthele nobilis

0,34a

Hi Sm trng

0,36a

Hi Sm en

5
6
7
8
9

Hi sm da trn
Hi Sm trng
Hi Sm nu
Hi Sm
Hi Sm xanh

Holothuria scabra
Holothuria martensii
Semper
Botradschia argus
Holothuria scabra
Actynopyga echintes
Holothuria sp.
Bahadschia graeffei

a. % so vi trng lng ti
b. % so vi trng lng kh

653

0,58a
0,36a
1,25b
2,10b
1,58b
3,05b

 Thnh phn v hm lng axt bo

Bng 1: Mu hi sm vng
TT

Axt bo

1
2
3
4

C14:0
C16:1(n-7)
C16:0
C18:1(n-9)

5
6

C18:0
C20:4 (n-6)

C20:5(n-3)

8 C20:1(n-9)
9 C20:0
10
11

Tn khoa hc

Tn thng

Thi gian
lu (Rt)
8,52
10,33
10,52
12,54

Tetradecanoic axit
Myristic
9-Hexadecenoic axit
Palmitoleic
Hexadecanoic axit
Palmitic
Cis
9-Octadecenoic Oleic
axit
Octadecanoic axit
Stearic
12,4
5,8,11,14Arachidonic
13,61
Eicosatetraenioic axit
5,8,11,14,17
EPA
13,67
Eicosapentaenoic axit
11-Eicosenoic axit
13,92
Eicosanoic axit
14,15
Cyclotetracosane
17,48
Others
Tng cc axt bo no:
Tng cc axt bo khng no:

Hm
lng [%]
0,89
1,89
14,58
24,9
9,2
5,9
4,24
4,68
2,21
1,93
39,64
17,68
43,28

Bng 2: Hi Sm trng (Holothuria scabra)

TT

Axt bo

Tn khoa hc

1
2
3
4
5
6
7
8

C14:0
C15:0
C16:1(n-7)
C16:0
C17:0
C18:1(n-9)
C18:0
C20:4 (n-6)

C20:5(n-3)

10
11
12
13
14

C20:1(n-9)
C20:0
C21:0

Tetradecanoic axit
Pentadecanoic axit
9-Hexadecenoic axit
Hexadecanoic axit
Heptadecanoic axit
Cis 9-Octadecenoic axit
Octadecanoic axit
5,8,11,14Eicosatetraenioic axit
5,8,11,14,17
Eicosapentaenoic axit
11-Eicosenoic axit
Eicosanoic
Heneicosanoic
Cyclotetracosane

Tn thng Thi gian

lu (Rt)
Myristic
8,52
9,162
Palmitoleic
10,33
Palmitic
10,52
Margnic
11,12
Oleic
12,18
Stearic
12,40
Arachidonic
13,61
EPA

13,68

2,61

13,92
14,16
15,16
17,48

9,77
2,54
2,54
3,07
56,16
17,55
26,29

Others
Tng cc axt bo no:
Tng cc axt bo khng no:

654

Hm
lng [%]
1,83
0,93
2,84
5,29
0,21
5,56
4,21
5,51

Bng 3: Mu Hi sm v (Microthele nobilis)

TT

Axt bo

Tn khoa hc

Tn thng

1
2
3
4
5
6
7
8

C14:0
C15:0
C16:1(n-7)
C16:0
C18:2(n-6)
C18:1(n-9)
C18:0
C20:4 (n-6)

Myristic

C20:5(n-3)

10
11
12
13
14

C20:1(n-9)
C20:0
C21:0

Tetradecanoic axit
Pentadecanoic axit
9-Hexadecenoic axit
Hexadecanoic axit
9,12-Octadecadienoic axit
Cis 9-Octadecenoic axit
Octadecanoic axit
5,8,11,14Eicosatetraenioic axit
5,8,11,14,17
Eicosapentaenoic axit
11-Eicosenoic axit
Eicosanoic
Heneicosanoic
Cyclotetracosane

Palmitoleic
Palmitic
Linoleic
Oleic
Stearic
Arachidonic
EPA
-

Thi
gian
lu (Rt)
8,52
9,54
10,33
10,52
12,13
13,18
12,40
13,61

Hm
lng[%]

13,67

4,02

13,62
14,16
15,15
17,48

12,43
3,44
1,65
2,77
26,52
30,63
42,75

Others
Tng cc axt bo no:
Tng cc axt bo khng no:

3,14
1,16
10,35
16,98
0,86
5,39
7,40
4,96

Bng 4: Mu Hi sm en (Holothuria vagabunda)

Thi gian
lu (Rt)

Hm
Lng[%]

8,52
9,53
10,33
10,52
11,17
11,99

2,97
2,37
11,3
10,15
1,45
1,32

9,12-Octadecadienoic axit Linoleic

12,13

0,67

Cis 9-Octadecenoic axit

Octadecanoic axit
Nonadecanoic

Oleic
Stearic

12,18
12,4
13,28

5,39
5,67
0,54

Arachidonic

13,61

8,99

EPA

13,68

8,55

13,92
14,16

8,55
2,02

TT

Axt bo

1
2
3
4
5
6

C14:0
C15:0
C16:1(n-7)
C16:0
C17:0
C18:3

Tetradecanoic axit
Pentadecanoic axit
9-Hexadecenoic axit
Hexadecanoic axit
Heptadecanoic axit
Octatrienoic

C18:2(n-6)

8
9
10

C18:1(n-9)
C18:0
C19:0

11

C20:4 (n-6)

12
13
14

Tn khoa hc

Tn thng

5,8,11,14- Eicosatetraenioic axit

C20:5(n-3) 5,8,11,14,17
Eicosapentaenoic axit
C20:1(n-9) 11-Eicosenoic axit
C20:0
Eicosanoic

655

Myristic
Palmitoleic
Palmitic
Margnic

15

C21:0

Heneicosanoic

16

C22:6(n-3)

4, 7, 10, 13, 16, 19

Docosahexaenoic axit
Cyclotetracosane

17

DHA
-

15,15

0,82

15,5

1,13

17,48

1,59

18 Others

37,78
Tng cc axt bo no:

21,02

Tng cc axt bo khng no:

41,2

Bng 5: Mu Hi sm da trn (Botradschia argus)

TT

Axt bo

Tn khoa hc

1
3
4
5
6
7

C14:0
C15:0
C16:1(n-7)
C16:0
C17:0
C18:3
C18:2(n-6)

Tetradecanoic axit
Pentadecanoic axit
9-Hexadecenoic axit
Hexadecanoic axit
Heptadecanoic axit

8
9
10
11

C18:1(n-9)
C18:0
C19:0
C20:4 (n-6)

12

C20:5(n-3)

13
14
15
16
17

C20:1(n-9)
C20:0
C21:0
C22:6

18
19

Tn thng
Myristic
Palmitoleic
Palmitic
Margnic

Thi gian
lu (Rt)
8,52
9,53
10,33
10,54
11,47
11,99
12,13

Hm
lng[%]
3,56
1,41
5,17
11,88
0,92
0,55
0,77
5,08
6,44
0,59
7,51

9,12-Octadecadienoic
axit
Cis 9-Octadecenoic axit
Octadecanoic axit

Linoleic

5,8,11,14Eicosatetraenioic axit
5,8,11,14,17
Eicosapentaenoic axit

Arachidonic

12,18
12,40
13,28
13,61

EPA

13,68

7,44

11-Eicosenoic axit
Eicosanoic
Heneicosanoic
4, 7, 10, 13, 16, 19
Docosahexaenoic axit
Cyclotetracosane

13,75
13,92
14,15
15,14
15,49

1,07
7,45
2,80
1,08
2,01

17,47

2,55
31,59
28,68
39,73

Oleic
Stearic

DHA

Others
Tng cc axt bo no:
Tng cc axt bo khng no:

656

c) Hoocmon (Xc nh hm lng cc hocmon testosteron, oestradiol, progesteron, prolactin

theo phng php min dch enzym Elisa).

Hm lng cc hoocmon steroit trong Hi sm (ng/g vck):

Mu
Hi Sm
(Holothuria sp,)
Hi sm nu
(Actynopyga
echinities)
Hi Sm da trn
(*) (H, argus)
Trung bnh

Testosteron
(ng/g vck)

Estradiol
(ng/g vck)

Progesteron
(ng/g vck)

Prolactin
(ng/g vck)

Oestradiol
(ng/g vck)

19,156

0,974

>219,85

<64 nM/l

21,130

0,785

>194,29

<64nM/l

0,97

20,143

0,879

>207,07

<64nM/l

0,970

(*): cha carotenoid-225g

Testosteron: Hooc mn sinh dc nam
Progesterol: Hooc mn sinh dc n

d) Hm lng nguyn t a vi lng v cacbuahydrat (Phn tch thnh phn cc

nguyn t a vi lng bng phng php nhiu x hunh quang tia X, xc nh cacbuahydrat
theo phng php so mu s dng anthrone)

Tn loi
Nguyn t
K (%vek)
Ca (%vek)
Fe (mg/kg vek)
Cu (nt)
Zn (nt)
Mn (nt)
Br (nt)
Co (nt)
Mo (nt)
Se (nt)
Cacbuahydrat
(% mu kh)

Holothuria
scabra

Actynopyga
echinites

Holothuria
sp,

Bohadschia
graeffei

2,879 0,42
8,0420,84
112,1514,42
22,586,63
21,966,26
<33,178
70,598,19

1,0553,1
<5,519
2,25

1,79 0,207
1,20 0,127
184,220,60
21,784,01
13,655,68

265,8427,80
<10,407
<3,3880,47
11,641,90
3,28

0,0340,009
20,9612,192
197,5520,89
62,596,70
63,386,74

381,3439,70
31,523,39

2,860,86
4,40

4,0670,446
1,1640,126
731,6379,22
125,6116,93
152,6116,90
246,6127,89
229,1924,80

7,170,65
13,692,34
3,90

Cc kt qu nghin cu ca chng ti cho thy:

a) Hm lng protein, lipit, cacbuahydrat trong cc loi Hi sm:
Hm lng protein tng s ca 2 loi hi sm Holothuria scabra v
Holothuria sp. l nh nhau v cao nht (67,82%), sau l ca Actynopyga
Echinites v Bohadschia graeffei. Trong khi hm lng cacbuahydrat ca
chng dao ng trong khong t 2,25% n 4,40%. Hm lng lipit ca
Bohadschia graeffei l cao nht v bng 3.05%, cn gi tr tng ng ca
Bohadschia scabra l thp nht (1,25%). Cc mu ti ca mt s loi hi sm
vng bin Nha Trang c hm lng lipit dao ng trong khong 0,27-0,58%.

657

b) Thnh phn v hm lng axt amin trong cc loi Hi Sm

Tht hi sm c y cc axt amin, c bit cc axt amin khng thay th.
Hi Sm (Holothuria sp.) v Hi Sm nu (Actynopyga echinitess) c hm
lng lizin, phenylAlanin, lxin + Izolxin cao hn Hi Sm trng (Holothuria
scabra) v hi sm xanh (bohadschhia graeffei). Ring hi sm nu c hm
lng lysin cao nht (9.87%).Hi Sm c hm lng Alanin (3.80%) v
tyrosine (3.05%) cao nht trong 5 loi hi sm. C 5 loi hi sm u c hm
lng cao cc axt amin: prolin, axit glutamic v theonine. Chc nng ca cc
axt amin c khng nh: Lizin, Lxin, Izolxin c tc dng tng trng c
th, ring lizin c nh hng n trng thi ca h thn kinh, hm lng kali
trong m xng, tng hp hemoglobin v s to thnh axt nucleic. Tyrosine l
nguyn liu tng hp sc t melanin. Khi c th thiu melanin s b bnh
bch tng, s nh sng, git nhn cu.. PhenynAlanin trong c v gan bin thnh
tyrosine nh enzim phenynalanin hydroxylaza.
c) Thnh phn v hm lng axt bo trong cc loi hi sm
Thnh phn v hm lng cc axt bo tn ti trong lipit mi loi i tng
nguyn liu t nhin, s quyt nh n bn cht v gi tr thc tin ca loi
nguyn liu . Trong i tng sinh vt bin, ngoi s c mt cc axt bo thng
dng thng c trong cc nguyn liu t nhin trn cn nh: Myristic C14:0,
Palmitic C16:0, Stearic C18:0, Oleic C18:1, Linoleic C18:2 v Arachidonic C20:4
(AA); cn c s tham gia ca cc axt bo khng no mch di vi hot tnh sinh
hc cao, c th ring ch c trong sinh vt bin l Eicosapentaenoic C20:5
(EPA) v Docosahexaenoic C22:6 (DHA).
Thnh phn v hm lng cc axt bo trong cc loi hi sm ni chung
u c cha cc loi axt bo ph dng trong thnh phn sinh vt bin, c 5 loi
u c hm lng cc axt bo khng no, a ni i tng i cao (26,2943,28%), v c 5 loi u c mt axt bo rt quan trng trong c th sng l AA
vi hm lng (4,96-8,99%) v EPA (2,61-8,55%). Tuy nhin DHA ch c mt
trong hi sm en (1.13%); Hi sm da trn (2.01%);
d) Thnh phn v hm lng cc nguyn t vi lng trong cc loi hi sm
Tt c 5 loi hi sm nguyn cu u giu st, m st l nguyn liu cn
cho s to mu ca c th. Trong hi sm v hi sm xanh li giu Cu v Zn,
m cc nguyn t ny c tc dng tt n chc nng sinh dc ca c th, tng
cng to tinh trng nam gii, pht trin bung trng n gii. Cu v Zn cn
gip lm tng tr nh v tc dng tt ln h thn kinh. Brm c tc dng an thn
lm cho gic ng c tt, u thy c hm lng cao hi sm nu v hi sm
(hi sm nu 265,84 27,8 mg/kg vek, hi sm 381,34 39,7 mg/kg vek),
trong khi hi sm trng ch c 70,598,1 mg/kg vek. iu l th l Selen l
nguyn t c lin quan n qu trnh chng xy ha, chng lo ha li c nhiu
nht trong hi sm nu (11,64 1,8mg/kg vek) v hi sm xanh (13.6 2,3 mg/kg
vek) nhiu gp i so vi hi sm trng (<5,519 mg/kg vek).
Nh vy qua cc kt qu nghin cu v thnh phn v hm lng cc axt
amin, cc nguyn t vi lng c th gii thch ti sao con ngi li dng hi
sm lm thc phm b dng c th v hi sm cha tp hp cht c tc dng b
mu, tng cng trao i cht b thn kinh, b m, trng dng, tr ha c th.

658

e) Hm lng cc hoocmon trong hi sm

Mt iu rt l th l pht hin thy trong hi sm c hm lng
Testosteron tng i cao (20,143 ng/g) nhiu gp ba ln so vi trong yn so
(8.54ng/g). Hm lng Progesteron nhiu gp hai ln yn so (Hi sm
207,7ng/g, yn so 100,63 ng/g). Trong Hi Sm cc hoocmon Estradiol v
Prolactin c hm lng khng ng k. Nh chng ta bit, Testosteron lm
pht trin cc du hiu sinh dc ph, to ra dc tnh nam gii. Th nghim kinh
in tim cho g trng thin Testosteron, mo g li pht trin, lng tr nn sc
s tr li, hung hng v p mi c, minh ha tc dng k trn ca
Testosteron. Testosteron nh hng n qu trnh chuyn ha ca nit v photpho
trong c th, n c tc dng kch thch qu trnh sinh tng hp protit gi nit, lm
gim bi tit ur, lm pht trin cc c. Kt qu ny gip ta c c s khoa hc
hiu c tc dng ca hi sm i vi kh nng sinh dc ca nam gii v tng
cng sc khe cho ngi gi. Qua kt qu phn tch trn cn thy hoocmon sinh
dc n trong hi sm cng kh cao, nh vy trong hi sm khng nhng c tc
dng kch dng m cn b m, rt tt vi ph n trong thi k mang thai, bi v
Progesteron c tc dng lm cho hp t bm chc vo nim mc t cung, chng
co bp t cung trnh sy thai...
XII.1.4. Nghin cu thnh phn cc cht c hot tnh sinh
hc ca mt s loi hi sm
Trong khun kh ti cp nh nc KC09.15, chng ti tin hnh
nghin cu xc nh thnh phn ha hc v hot tnh sinh hcca loi hi sm
Holothuria vagabunda, v H. martensii.
Cc mu hi sm c s dng nghin cu:
Tn Vit Nam: Hi sm en
Tn khoa hc: Holothuria vagabunda
a im ly mu: Vng bin Qung Ninh.
Ni sng: Khu vc Bi Hng Vn n hn
Khe Tru. Hi sm en thng sng trn cc
rn san h v theo nc triu tp vo bi ct
kim n.
Ngi gim nh: TS. Cng Thung

Vin Ti nguyn v Mi trng Bin

Tn Vit Nam: Hi sm en
Tn khoa hc: Holothuria martensii
a im ly mu: Vng bin Qung Ninh.
Ni sng: y ct th, v sinh vt ln cn, rn
v rn san h hoc thm c bin.
Ngi gim nh: TS. Cng Thung

Vin Ti nguyn v Mi trng Bin

659

Kt qu ng ch l chng ti phn lp v xc nh c t c hai

hoi hi sm ny 02 hp cht tritecpen saponin dng khung holosthan l:
holothurin A va holothurin B vi hm lng kh cao. Chng l cc hp cht c
cu trc ha hc rt c o v th hin nhiu hot tnh sinh hc ng quan tm.
Cc nghin cu nh gi hot tnh sinh hc cho thy: cc hp cht ny u th
hin hot tnh gy c t bo cao i vi nhiu dng t bo ung th ngi nh KB
(ung th biu m), FL (ung th mng t cung), Hep-G2 (ung th gan). Ngoi ra,
hp cht holothurin A cn th hin hot tnh khng MAO mnh (IC50= 1,5
g/ml).
Mt s hp cht in hnh phn lp c t cc loi hi sm:
21

HO

O
18

12

14

9
8

5'

O O

OH

27

OH

16
15

24
26

17

13

19

23
20

28
7
6

30

29

1'
2'

N aO 3S O
O

CH3
5 ''
6'''

OH

C H 2O H
5'''

O O

2 ''
1'''

OH

2 '''

OH

Holothurin A (C54H86NaO27S)
Ngun gc: Hi sm en
Tn khoa hc: H. vagabunda,
H. martensii
Cht thu c dng bt mu trng
Nhit nng chy: 228-230oC
Hot tnh gy c t bo:
KB (IC50=0,95 g/ml)
Hep-G2 (IC50=2,15g/ml)
FL (IC50=1,10g/ml).
Hot tnh khng MAO
(IC50= 1,5 g/ml).

C H 2O H

OH

5'' ''
7 '' ''

O
1''

OCH3
2 ''

OH

OH
21

O
HO
13

14

9
8

10
3

O
5'

OH

27

15

7
6

30

OH

16

25

28

26
22

17

11

24

20

18

12
19

23

29

1'
2'

NaO 3 SO
O

6"

CH 3
O

5"

OH
OH

Holothurin B (C41H63NaO17S)
Ngun gc: Hi sm
Tn khoa hc: H. vagabunda,
H. martensii
Cht dng bt mu trng.
Nhit nng chy: 251-253oC
Hot tnh gy c t bo:
KB: IC50 = 1,5g/ml
Hep-G2: IC50 = 1,15g/ml
FL: IC50 = 0,5g/ml

1"
2"

OH

Trn c s cc kt qu nghin cu trn, chng ti ng k v c

cp 01 bng c quyn sng ch: Phng php chit hp cht holothurin B t
loi hi sm Holothuria vagabunda, s 5846, ngy 23/08/2006 ti Cc S hu
Tr tu, B Khoa hc v Cng ngh Vit Nam.

660

XII.1.5. C s hnh thnh sn phm

T cc kinh nghim c truyn cng vi s minh chng bng cc nghin
cu hin i v thnh phn ha hc v hot tnh sinh hc cho thy, hi sm l
loi ng vt c gi tr dinh dng rt cao vi hm lng di do ca cc loi
protein, lipit, cc axit bo, cc axit amin khng thay th, cc loi vitamin, khong
cht, cc nguyn t vi lng.... Mc d cc nguyn t vi lng tn ti trong c th
vi khi lng v cng nh nhng n ng vai tr quyt nh trong hu ht cc
phn ng sinh ha v cc qu trnh trao i cht trong c th. Vi hm lng cc
nguyn t vi lng kh y v di do trong hi sm lm cho n tr thnh
mt loi thc phm c gi tr rt cao.
Hn th na hi sm cn cha nhiu loi hoocmon quan trng, c bit l
testosteron vi hm lng (20,143 ng/g) nhiu gp 3 ln trong yn so (8,45
ng/g), mt hooc mn c bit n c vai tr quan trng trong vic pht trin
cc du hiu sinh dc ph, to ra dc tnh nam gii. Testosteron nh hng n
qu trnh chuyn ha ca nit v photpho trong c th, n c tc dng kch thch
qu trnh sinh tng hp protein, gi nit, lm gim bi tit ur, lm pht trin cc
c. Kt qu ny gip ta c c s khoa hc hiu c tc dng ca Hi Sm i
vi kh nng sinh dc ca nam gii v tng cng sc khe cho ngi gi.
Cc nghin cu v thnh phn ha hc cho thy hi sm cha rt nhiu
cht c cc hot tnh qu bu nh hot tnh gy c t bo, chng xy ha, khng
MAO. c bit l s c mt ca cc hp cht holothurin c hot tnh gy c t
bo rt mnh i vi nhiu dng t bo ung th ngi, ha hn cho vic pht trin
cc dc phm c tc dng phng v tr bnh ung th. Nh vy hi sm khng
nhng c gi tr dinh dng rt cao m cn c tc dng phng v cha bnh.
T cc nghin cu ca chng ti, cng vi vic ngun nguyn liu hi sm
c th c ch ng m bo bng cc phng php nui trng, t c s
khoa hc vng chc cho vic xy dng, sn xut v pht trin mt loi thc phm
chc nng c gi tr cao l Vin nang mm hi sm: HASAMIN. iu ny s
gp phn nng cao tnh n nh v gi tr dinh dng cng nh gi tr dc liu
ca hi sm, gp phn s dng ngun nguyn liu qu gi ny mt cch hp l v
c hiu qu nht .

661

XII.3. kt qu v tho lun

XII.3.1. xy dng quy trnh cng ngh sn xut vin nang
mm hi sm Hasamin
Cng thc pha ch cho mt m sn xut 100.000 nang:
- Bt hi sm
20 kg
- Du u nnh
17 kg
- Du c
12 kg
- Sp ong trng
19 kg
- Gelatin
9 kg
- Glycerin
3 kg
- Dung dch Sorbitol 70%
1 kg
- Nipagin
0,1 kg
- Nipasol
0,03 kg
- Etyl vanilin
0,1 kg
- Titan dioxit
1 kg
- Mu Brown R
0,1 kg
- Nc ct
16 lt
- Etanol 90%
0,5 lt
XII.3.1.1. Chun b nguyn ph liu, my mc, con ngi
a ph liu:
- Mng nhm, PVP t tiu chun c s
- Hp giy ng quy cch
- Hm carton 3 lp ng quy cch
b. Dng c my mc:
Phng lm vic, my mc, dng c cn, ong, pha ch phi kh, sch,
khng ln mi l, chng nhim cho. Cc thit b phi m bo an ton v in.
c. Nguyn liu:
Lnh nguyn liu theo nh mc k thut. Khi lnh nguyn liu phi c 02
ngi trong c mt cn b k thut kim tra cht lng v s lng nguyn
liu, i chiu phiu xut kho, phiu kim nghim nguyn liu.
d. Con ngi
Ngi pha ch phi thc hin ng quy ch v v sinh v an ton lao ng.
XII.3.1.2. Quy trnh bo ch
a. cn: Cn nguyn liu theo cng thc pha ch. Ghi khi lng tng loi vo h
s l sn xut
b. Bo ch hn hp v nang.
- Trn gelatin vi 15 lt nc ct, glycerin, dung dch sorbitol 70% un
nng 70oC trong 3-4 gi.
- Ha mu brown R, titan dioxit vo 500 ml nc ct, un nng 60oC,
lc qua ry s 180, cho hn hp vo dung dch gelatin trn, khuy trn u.
- Ha tan etyl vanilin vo 0,5 lt nc ct, un nng 70oC cho tan, ri
cho vo khi gelatin trn.
- Ha tan nipagin, nipazol vo 0,5 lt etanol 90%, cho vo khi gelatin
trn.
662

Trn cho ti khi to thnh mt mu ng nht. Ht chn khng kh bt

trong 1 gi. Bo qun khi gelatin to v nang nhit 60oC trong 8 gi.
c. Pha ch hn hp thuc
- Cho du c vo du u nnh, un nng nhit 70oC, khuy trn
trong thi gian 5 pht (hn hp 1).
- Nghin mn bt hi sm, ry thng qua ry s 180.
- un nng chy sp ong trng.
- Phn tn bt hi sm vo sp ong trng, khuy trn u trong thi gian
05 pht (hn hp 2).
- Cho hn hp 1 vo hn hp 2, khuy trn u to thnh hn hp thuc
ng nht (hn hp 3).
- Cho hn hp 3 qua my xay keo (hn hp thuc)
- Ht chn khng, kh bt hn hp thuc. Bo n hn hp thuc nhit
50oC, thi gian 2 gi.
d. Kim tra bn thnh phm
Hn hp thuc v hn hp v nang c kim tra bn thnh phm v cc
ch tiu sau:
- ng u ca khi mu gelatin.
- ng nht ca hn hp thuc.
- nh lng cc thnh phn ca thuc theo tiu chun c s. Xc nh khi
lng thuc cn ng vo mt nang. Ghi kt qu kim nghim bn thnh phm
vo h s l sn xut.
e. ng nang
Sau khi kim tra bn thnh phm, hn hp v nang v hn hp thuc c
chuyn sang cng on ng nang.
My ng nang c v sinh sch s, kim tra an ton v in. My c
lp khun theo ng chng loi nang cn ng (vin nang mm hnh Oblong 16).
Tin hnh ng th thuc vo nang theo kt qu kim tra bn thnh phm.
Sau khi kim tra v nang t yu cu th tin hnh ng thuc vo nang.
Trong qu trnh ng nang, c 15 pht kim tra khi lng trung bnh nang mt
ln, thng xuyn kim tra dy ca v nang v kn ca nang.

Hnh XII.3.1.2. nh my ng nang

663

f. Sy nang
Cho cc vin nang mm i qua mt my sy trn v tin hnh qu trnh
sy. Tri cc vin nang mm sy ln khay, nhit phng (20-24oC) v
m tng i 20-22% trong 12-24 gi cho n khi cc vin nang mm n nh
hon ton. Nang t yu cu c bo qun trong ti PE.
g. Chn nang
Loi b nhng nang khng t tiu chun v hnh thc v cht lng.
h. p v
Cho my chy th v iu chnh giy nhm, PVC vi khun my cho khp
nhau. Cho my chy sn phm t yu cu, tip tc cho my chy n khi ht
vin. V thuc p xong phi m bo bm dnh PVC, s l sn xut, hn dng
r rng mi em ng hp. Kim tra kn ca v thuc 1 gi/ln theo tiu chun
c s bao b.
i. ng gi
V 5 vin, hp 6 v; l 100 vin. ng thng carton.
j. Nhp kho
- Nhng v thuc khng t tiu chun v hnh thc, khi lng, bm
dnh, nang mp mo u c tp trung li em x l cho vo l m tip theo.
- Nhng nang v, bn, dnh du m, nang v chy phi loi b.
- Kim nghim thnh phm t tiu chun chuyn sang nhp kho.
k. Thnh phm
- Mi vin nang thnh phm c cha:
- Bt hi sm
200 mg
- Du u nnh
170 mg
- Du c
120 mg
- Sp ong trng
190 mg
- Gelatin
90 mg
- Glycerin
30 mg
- Dung dch Sorbitol 70%
10 mg
- Nipagin
0,1 mg
- Nipasol
0,03 mg
- Etyl vanilin
1 mg
- Titan dioxit
10 mg
- Mu Brown R
1 mg
- Ch nh
Tng cng sinh lc, duy tr trng thi hot ng cao, chng stress, chng
lo ha, tng cng min dch, tng sc khng ca c th, b sung axit amin,
cc nguyn t vi lng, tng tun hon mu, ci thin kh nng hp th xy,
chng mi c tim, thc y qu trnh chuyn ha v hp th, tng sinh tng hp
protein, gim cholesterol mu.
- Cch dng v liu dng: Ung mi ln 2 vin, ngy 2 ln.
- Bo qun, hn dng: Bo qun ni kh ro, nhit 15-25oC, hn dng 24
thng k t ngy sn xut

664

Du u nnh,
du c

Bt hi sm,
Sp ong
trng

Cn

Cn

Hn hp 1

Hn hp 2

Hn hp thuc

Gelatin, glycerin,
dd Sorbitol, nc
ct

Cht mu,
titan dioxyt,
nc ct

Ethyl
vanilin, nc
ct

Cht bo
qun, etanol
90%

Cn

Cn

Cn

Cn

Kim nghim
bn thnh
phm

Hn hp v nang

ng nang

Sy nang

Chn nang

p v

ng gi
Kim nghim
thnh phm
Thnh phm,
Nhp kho
S quy trnh sn xut Vin nang mm hi sm: HASAMIN

665

XII.3.2. Nghin cu xc nh Thnh phn, th nghim c

tnh cp, c tnh bn trng din v tc dng trn ng
vt ca sn phm vin nang mm hi sm Hasamin
XII.3.2.1. Xc nh thnh phn vin nang mm HASAMIN
a. Xc nh thnh phn dinh dng, ch tiu ha l, vi sinh ca sn phm
Sau khi xc nh c cc iu kin cng ngh ca quy trnh sn xut
vin nang mm hi sm HASAMIN, vic sn xut c tin hnh. Sn phm
to ra c phn tch cc ch tiu cht lng v kt qu c trnh by di y.
Bng XII.3.2.1.a. Cc ch tiu vi sinh vt
TT
1
2
3
4
5
6
7

Ch tiu
Tng s vi khun hiu kh
(CFU/g)
E. coli (37oC/96h)
(CFU/g)
Coliforms (37oC/48h)
(MPN/g)
Cl. perfringens (37oC/24h)
(CFU/g)
B. cereus (37oC/24h)
(CFU/g)
S. aureus (37oC/48h)
(CFU/g)
Salmonella (37oC/48h)
(CFU/g)

Phng php

Kt qu

TCVN 51651990
TCVN 68452001
TCVN 48822001
TCVN49911989
TQBYT
3550-2001
TCVN 48301989
TCVN 48291989

7,5 101

Gii hn ti a
theo TCCS
104

10

10

10

Bng XII.3.2.1.b. Cc ch tiu ha l

TT

Ch tiu

Phng php

Kt qu

m (%)

TCVN 5701-1993

Nng lng (Kcal/100g)

AOAC-1997

351,2

Hm lng protein (%)

NMKL.No6

7,43

4
5

Hm lng gluxit (%)

Hm lng lipit (%)

0
34,7

Hm lng tro tng s (%)

7
8

Hm lng tro khng tan

trong HCl (%)
Hm lng Ch (mg/kg)

TCVN 4594-1990
KNLTTP-NXBYH
1975
KNLTTP-NXBYH
1975
KNLTTP-NXBYH
1975
AOAC-2000

khng pht hin

Hm lng Asen (mg/kg)

AOAC-2000

khng pht hin

666

0,5

2,82
0,82

Sn phm c xy dng tiu chun c s v c B Y t cp chng

nhn tiu chun sn phm s 4978/2006/YT-CNTC vi tn gi : Thc phm chc
nng Vin nang mm hi sm HASAMIN.
- Trng thi: dng vin nang mm
- Mu sc: mu nu en
- Mi: mi thm, hi tanh c trng ca sn phm
- V: c v mn
- tan r: khng qu 30 pht
- ng u khi lng: 7,5% so vi khi lng trung bnh thuc trong
nang

Hnh XII.3.2.1.a. Sn phm nang mm hi Sm HASAMIN

v chng nhn tiu chun sn phm
667

b. Xc nh thnh phn axit amin trong vin Hasamin

Qua kt qu nghin cu trong bng XII.3.2.1.a cho thy hm lng cc
axit amin c trong sn phm vin nang Hasamin c thnh phn axit amin tng
i cao khng km g so vi cc sn phm khc trong nc v ngoi nc nh
ru Hi m, vin Taxaton, vin Triplex hay gi khang thai ca Trung Quc.
sn phm gi Khang thai, thnh phn c 18 axit amin v hm lng cng tng
i cao nh: Histidin: 9,88; Xistein: 9,54; Lxin: 8,21; A.Aspactic: 7,22, Trong
khi vin tng lc Hasamin thnh phn cng c ti 17 axit amin v hm
lng cng rt phong ph nh: A.Aspactic: 2,87; A.glutamic: 3,97; Treonin:
2,96; Xerin: 3,09; Glyxin: 5,98; Tirozin: 3,12; Alanin: 3,26; Acginin: 6,24;
Histidin: 2,98; Valin: 3,10; Metionin: 3,12; PhenylAlanin: 4,06; Izolxin: 5,12;
Lxin: 4,80; Lizin: 1,12; 4-Hydroprolin: 4,38; Prolin: 0,98,
Bng XII.3.2.1.a: Hm lng cc axit amin trong vin HASAMIN so vi cc sn
phm khc (n v tnh:%g a.amin/100g mu).
Tn axit amin

A,Aspactic
A,glutamic
Treonin
Xerin
Glyxin
Tirozin
Alanin
Acginin
Histidin
Valin
Metionin
PhenylAlanin
Izolxin
Lxin
Lizin
4-Hydroprolin
Xistein+Xistin
Prolin

Vin
HASAMIN

Ru Hi
M (VN)

Vin
Taxaton
(VN)

Gi
Khang
Thai (TQ)

Vin
Triplex
(TQ)

2,87
3,97
2,96
3,09
5,98
3,12
3,26
6,24
2,98
3,10
3,12
4,06
5,12
4,80
1,12
4,38
0,98

3,24
2,50
0,12
1,12
3,70
0,38
0,52
1,48
1,26
2,01
1,18
1,05
2,01
1,53
0,85

3,03
2,80
1,22
4,68
6,96
7,50
3,50
3,10
3,87
7,80
2,82
2,90
4,85
2,55
7,20
5,80
6,50
0,12

7,22
6,15
4,02
3,11
6,28
4,02
4,07
5,78
9,88
4,20
6,19
5,44
3,19
8,21
3,15
7,14
9,54
7,89

4,12
6,05
2,80
2,18
10,88
2,90
5,92
4,03
3,99
1,80
1,49
3,49
2,77
3,12
4,28
3,60
4,82

* Gi tr thng k vi : (n= 15, p 0,05)

Chng ta u bit cc axit amin ng vai tr rt quan trng v thit yu

trong c th. Mi axit amin u c nhng ng gp tng chc nng cho c th
nh: A.Aspactic l axit amin c tc dng gip cho qu trnh gii c v tng
cng sc bn c th. A.glutamic b no, gip mau lnh vt thng, kch thch
hng phn thn kinh. Tirozin tng cng sinh lc, hi phc sc kho, b gan.
Alanin n nh ng trong mu. Acginin l axit amin thit yu c tc dng tng
cng h thng min dch, tng trng hoocmn gip cho ngn nga v iu
chnh cc chng bt lc ca nam gii.

668

Hnh XII.3.2.1.b. th axit amin c trong vin tng lc HASAMIN

Histidin c tc dng b khp, tng to mu chng d ng. Metionin, Valin
gip cho n nh tm l. PhenylAlanin c tc dngngn nga chng bo ph,
gim trm cm, gim au nhc do bnh cm, kinh nguyt, gim vim khp.
Izolxin l axit amin thit yu n nh lng ng trong mu, tng cng h tr
pht trin c bp. Lxin lm tng c bp, n nh tm l. Lizin gip cho qu trnh
hnh thnh khng th, n nh ni tit t. Hydroprolin, Xistein tng cng min
dch chng xy ho, v hiu ho cc gc t do, gip cho mau lnh vt thng,
pht trin v lm p t bo da, tc. Prolin c tc dng gip cho qu trnh vn
ng khp v cc c chng, c tim. Treonin kch thch h thng thn kinh thc
vt gip cho ng ngon tng cng chc nng gan. Xerin tng cng min dch,
sc khng v tng sc bn. Glyxin tng c bp, gip tuyn tin lit kho mnh.
Vi thnh phn axit amin ca vin tng lc HASAMIN cho thy gi tr
chc nng dinh dng khng km g so vi cc sn phm tng t trong nc
v nc ngoi, do vy sn phm Hasamin dng cho vic nng cao th lc ca vn
ng vin l rt tt.
c. Nghin cu thnh phn cc cht vi khong trong vin Hasamin
V hm lng cc hot cht vi khong c trong vin Hasamin qua kt qu
nghin cu trong bng XII.3.2.1.b cho thy: Hm lng ng (Cu) hu c c
trong vin Hasamin l 3,73 mg/kg vck, St (Fe) l 19,89 mg/kg vck, km (Zn)
hu c l 8,20, hm lng km l 8,20 mg/kg vck v Mangan (Mn) l
37,19mg/kg vck. Nhn chung hm lng cc cht vi khong c trong vin tng
lc Hasamin so vi cc sn phm tng t trong nc cng nh so vi sn phm
gi khang thai v vin Triplex ca Trung Quc th kt qu cng khng khc nhau
nhiu. Thm ch mt s thnh phn ca hm lng nguyn t km hay mangan
(Mn) cn cao bng hoc hn.

669

Bng XII.3.2.1.b: Hm lng cc cht vi khong c trong vin Hasamin so vi

cc sn phm khc.
Hm lng vi
cht

Vin
Hasamin
(VN)

Ru
Hi M
(VN)

Vin
Taxaton
(VN)

Gi
Khang thai
(TQ)

Vin
Triplex
ca (TQ)

Cu(mg/kg vck)

3,73

4,12

4,01

6,22

3,92

Fe (mg/kg vck)

198,89

58,22

227,20

240,02

238,27

Zn (mg/kg vck)

8,20

2,67

6,26

11,28

3,24

Mn(mg/kg vck)

37,19

15,09

27,24

37,91

42,76

* Gi tr thng k vi : (n= 15, p 0,05)

Hnh XII.3.2.1.c. Biu hm lng cc cht vi khong c trong vin HASAMIN

Vi thnh phn cc cht vi khong hu c c trong vin tng lc Hasamin
rt phong ph nh nguyn t km c trong vin Hasamin cao s gip cho qu
trnh tng cng th lc, tng cng min dch gip c th thc hin tt chc
nng ni tit, tun hon mu, mau lnh vt thng, khng ch t bo gy nhiu.
c bit nguyn t km cn ng vai tr chc nng duy tr sinh l ho trong c
th, tng kh nng sinh dc cho nam gii, gim bt cc nguyn t vi lng t do.
kch thch cc hoocmn tuyn thng thn hot ng, st s gip cho qu trnh
tng to hng cu, hemoglobin trong c th rt tt. Hm lng ng, mangan c
trong vin Hasamin gip cho s pht trin tr tu, kch thch phn x thn kinh
c tt hn.
d. Nghin cu thnh phn cc hot cht hoocmn steroit trong vin Hasamin
Kt qu phn tch hm lng cc hot cht steroit c trong vin
HASAMIN trong bng XII.3.2.1.c l kh cao so vi cc sn phm khc, k c sn
phm ngoi.

670

Bng XII.3.2.1.c. Kt qu phn tch hm lng cc hot cht steroit c trong vin
Hasamin so vi cc sn phm khc.
Hm lng steroit
- Testosteron(mg/kg vck)
- Progesteron(mg/kg vck)
- Cortisol (mg/kg vck)

Vin
Hasamin
(VN)
62,17
65,24
17,53

Ru
Hi M
(VN)
11,02
9,28
8,22

Vin
Taxaton
(VN)
68,00
65,07
19,40

Gi
Khang
thai (TQ)
67,35
52,15
59,24

Vin
Triplex
(TQ)
32,08
38,72
55,12

* Gi tr thng k vi : (n= 15, p 0,05)

Hm lng Testosteron (mg/kg vck) c trong vin HASAMIN l (62,17),

v ru Hi m l (11,02), vin Taxaton (68,00) v gi khang thai ca Trung
quc (67,35) nhng cao hn vin Triplex ca Trung quc (32,08).
Hm lng Progesteron (mg/kg vck) ca vin HASAMIN l (65,24) cao
hn nhiu so vi cc sn phm trong nc v ca Trung Quc nh ru Hi m
(9,28), vin Taxaton (65,07), gi khang thai (52,15) v vin Triplex (38,72).
Hm lng Cortisol (mg/kg vck) ca vin HASAMIN l (17,53) cao hn
tt c cc sn phm trong nc nh ca ru Hi m (8,22), vin Taxaton (19,40)
v khng thua km g cc sn phm ca Trung Quc nh gi Khang Thai (59,24),
vin Triplex (55,12).
So vi mt s sn phm khc th vin Hasamin c thnh phn v hm
lng cc hot cht hoocmn steroit nh Testosteron, progesteron, cortisol cng
rt phong ph. Cc hoocmn steroit ny u c tc dng kch thch tng hp
lipoprotein gip cho vic b p v cn bng glucose v lipit trong mu.
Hoocmn steroit vi nng cao c tc dng tng cng min dch, chng vim
nhim, chuyn ho mui ln tuyn nc bt v tuyn m hi cng nh s vn
chuyn mui v nc qua nim mc ng tiu ho, tham gia vo s duy tr cn
bng nc v in gii m bo s vn ng tiu hao nng lng c th. Thnh
phn hot cht hoocmn steroit c trong vin HASAMIN s lm tng gi tr b
dng ca vin Hasamin.
XII.3.2.2. Nghin cu c tnh cp v c tnh bn trng din ca vin
nang mm hi sm HASAMIN
a. Khi lng trung bnh lng thuc ng trong vin nang.
Cn 20 nang, xc nh khi lng trung bnh lng bt thuc c trong mi
vin nang l 0,202g/ vin .
b. Th c tnh cp :
Mu th: Ct vin nang mm, p ra ht cht dch v thm nc, trn u
thu c hn dch c cha lng thuc l 0,1g/ml. Khuy trn u hn dch
mi khi ly cho chut ung.
Tin hnh: Chut c nhn n 15 gi trc khi th nghim, nc ung
bnh thng. Cho chut ung kim cong u t, a thuc vo thng d dy chut.
Thm d cc liu gy c trn ng vt th nghim.
Da trn kt qu thm d, th nghim tin hnh trn 60 chut, chia lm 6
nhm, mt nhm chng dng nc ct v 5 nhm th. Cc nhm th cho ung

671

hn dch mu th chun b trn t 1- 3 ln trong ngy, cch nhau 3 gi. Mt

nhm cho ung vi th tch ti a chut c th chu ng c (0,5ml/10g chut/
ln). Theo di chut 5 ngy sau khi cho ung. B tr th nghim v kt qu thu
c th hin trn bng XII.3.2.2.a.
Bng XII.3.2.2.a: B tr th nghim v kt qu th c tnh cp:
Nhm
1
2
3
4
5
6

Liu dng
(ml/ 10g chut)
0,2 ml x 1 ln
0,2 ml x 3 ln
0,3 ml x 3 ln
0,4 ml x 3 ln
0,5 ml x 3 ln
0,5 ml nc x 3 ln

Liu dng
(g / kg chut)
2,0 g
6,0 g
9,0 g
12,0 g
15,0 g
nc ct

S chut th
nghim
10
10
10
10
10
10

S chut cht
0
0
0
0
0
0

Nhn xt: Sau khi ung nc ct v mu th, chut tt c cc nhm u hot

ng, n ung bnh thng k c nhm ung 3 ln vi th tch ti a cho chut
th nghim (0,5 ml/ 10g chut/ln). Khng c chut no cht trong thi gian theo
di.
c. Th c tnh bn trng din
Mu th: Liu th: 2 vin/ kg/ ngy
Ct vin nang mm, p ra ht cht dch v thm nc, trn u thu c hn
dch c cha lng bt thuc tng ng 2 vin/ 5ml. Khuy trn u hn dch
mi khi ly cho th ung.
Chun b mu th hng ngy trc khi cho th ung.
B tr th nghim v tin hnh: Th nghim c tin hnh trn 20 th, chia
lm hai nhm chng v th, mi nhm 10 th.
- Nhm chng: ung nc ct 5,0ml/ kg/ ngy.
- Nhm th : ung dch mu th chun b nh trn, liu 0,4g/ kg/ ngy.
Cho th ung hng ngy, lin tc trong mt thng.
Th c 2 nhm c nui dng trong cng iu kin v cng ch
dinh dng.
Theo di v nh gi: Theo di th hng ngy v mc tiu th thc n,
kh nng hot ng, tnh trng phn, lng.
Trc khi th nghim, xc nh ch s bnh thng ca th v cn nng, cc
du hiu ton thn, cc ch s v huyt hc, sinh ho chc nng gan, thn. Ht
thi gian th nghim, nh gi li th trn cc ch tiu nh trc th nghim. M
th quan st i th cc t chc gan, thn, phi, tim v hnh dng, mu sc, th
cht, tnh trng xung huyt...
So snh kt qu trc v sau th nghim ca nhm chng, nhm th, gia
nhm chng v nhm th theo phng php thng k y sinh hc.
Kt qu:
- Tnh trng th: Trong thi gian th nghim, hu ht th c hai nhm
hot ng bnh thng, n ung tt, phn kh, lng mt; khng c hin tng
rng lng hoc kh cng.

672

- Quan st i th cc t chc: Khi kt thc th nghim, tin hnh m th,

quan st cc t chc gan, thn, tim, phi. Khng nhn thy biu hin khc thng
trn cc t chc ca th nhm th so vi nhm chng.
- Quan st m bnh hc trn tiu bn vi th: Phn tch m bnh hc c
thc hin ti Khoa Nghin cu thc nghim - Vin Y hc c truyn Qun i.
Hnh thi vi th gan sau ung thuc:
+ L chng: cu trc ca gan bnh thng.
+ L th nghim: cc b gan, t bo khng hoi t. Cc xoang mch
khng gin, khng xung huyt.
Hnh thi vi th thn:
+ L chng: Cc cu thn khng xung huyt
+ L th nghim: cu thn v ng thn bnh thng, khng thy thoi ho
biu m ng thn
Kt qu cn nng th v cc ch s huyt hc, ho sinh c ghi trn cc
bng XII.3.2.2.b-d.
Bng XII.3.2.2.b. Kt qu theo di cn nng th
Nhm
Chng
Th
P (chng-th)

n
10
10

Cn nng (kg)
Trc th nghim
Sau th nghm
2,12 0,12
2,15 0,12
2,14 0,13
2,16 0,14
0,329
0,027

P
(trc - sau)
0,026
0,031

Bng XII.3.2.2.c. Kt qu theo di cc ch s sinh ho

Ch tiu nh gi
Protein
ton phn
(g/ l)
Bilirubin
(mol/ l)
GOT
(UI/l)
GPT
(UI/l)
Ure
(mmol/l)
Creatinin
(mol/ l)

Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)

n
10
10
10
10
10
10
10
10
10
10
10
10

Trc
th nghim
48,2 3,5
55,3 4,3
0,152
3,82 0,41
3,78 0,35
0,457
51,3 6,5
46,5 8,9
0,167
88,5 9,7
89,6 12,1
0,467
6,18 0,45
5,38 1,12
0,151
114,2 12,5
108,1 10,6
0,078
673

Kt qu
Sau
th nghim
53,1 5,9
50,3 3,4
0,187
3,17 0,54
2,89 0,32
0,239
62,1 13,9
56,5 6,3
0,325
78,6 10,8
80,3 11,4
0,371
6,28 0,76
5,67 0,52
0,120
116,8 10,7
110,2 6,8
0,115

P
(trc- sau)
0,109
0,332
0,032
0,012
0,039
0,013
0,016
0,109
0,502
0,226
0,362
0,417

Bng XII.3.2.2.d. Kt qu theo di cc ch s huyt hc

Ch tiu nh gi

Hng cu
(x 1012/ l)
Bch cu
(x 109/ l)
Huyt sc t
(g/l)
Hematocrit
(%)

Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)
Chng
Th
P (chng-th)

n
10
10
10
10
10
10
10
10

Trc
th nghim
5,90 0,19
5,89 0,21
0,52
5,17 0,16
5,15 0,25
0,201
123,3 2,9
121,7 2,5
0,639
39,1 1,8
37,6 2,4
0,139

Kt qu
Sau
th nghim
5,88 0,17
5,96 0,28
0,013
5,16 0,14
5,19 0,19
0,026
114,3 3,0
113,1 2,5
0,954
37,8 2,1
36,5 2,2
0,176

P
(trc- sau)
0,194
0,036
0,450
0,016
0,035
0,019
0,152
0,116

Kt lun:
Mu vin nang HASAMIN, l SX: 110707 gi ti yu cu th c tnh cp v
bn trng din c cc kt qu nh sau:
- Th c tnh cp trn chut nht trng: cho ung hn dch mu th trong
nc t 1-3 ln cch nhau 3 gi trong ngy, vi tng liu t 2,0 15,0g/ kg
khng nhn thy biu hin ng c no trn chut th nghim trong thi gian theo
di (k c nhm chut ung vi liu ti a 0,3g/ 10g chut/ ln). Khng xc nh
c liu gy cht 50% ng vt th nghim (LD50) v khng tm c liu gy
cht chut mc d cho ung vi mc liu cao.
- Th c tnh bn trng din: cho th ung lin tc trong mt thng vi mc
liu 2 vin/kg/ ngy c cc kt qu nh nu trn. Cha nhn thy s khc
nhau ng k v tnh trng th, cc ch s huyt hc v sinh ho, i th cc tng
ph sau th nghim gia nhm chng v nhm th. Hnh nh tn thng m bnh
hc t chc gan v thn trn tiu bn vi th khng c s khc nhau nhiu gia
nhm chng v nhm th.
XII.3.2.3. Th nghim tc dng ca vin Hasamin trn ng vt thc
nghim
Kt qu nh gi cc ch tiu sinh hc tinh trng th trc v sau b sung
ch phm c trnh by bng XII.3.2.3.a-c.
Bng XII.3.2.3.a. Ch tiu sinh hc tinh dch th trc b sung vin Hasamin,
Placebo
Ch tiu theo di
Lng tinh dch (ml)
Hot lc tin thng (A %) ca tinh trng
Nng tinh trng (triu/ml)
Tng s tinh trng tin thng (V.A.C:triu/
mt ln ly tinh)
674

Trc b sung
vin Hasamin
0,56 0,11
65,86 5,14
250,15 16,42
106,27 15,73

Trc b sung
vin Placebo
0,58 0,10
66,77 3,27
248,05 16,74
110,45 13,79

T l k hnh (%) ca tinh trng

T l sng (%) ca tinh trng
Sc khng ca tinh trng i vi (NaCl 1%)
PH tinh dch

20,52 4,35
74,51 7,38
538 14,36
6,75 0,22

19,60 5,36
73,29 9,61
532 16,75
6,80 0,10

Bng XII.3.2.3.b. Ch tiu sinh hc tinh dch th sau 10 ngy b sung vin
Hasamin, Placebo
Ch tiu theo di
Lng tinh dch (ml)
Hot lc tin thng (A %) ca tinh trng
Nng tinh trng (triu/ml)
Tng s tinh trng tin thng (V.A.C:triu/
mt ln ly tinh)
T l k hnh (%) ca tinh trng
T l sng (%) ca tinh trng
Sc khng ca tinh trng i vi (NaCl 1%)
PH tinh dch

B sung vin
Hasamin
0,78 0,17
72,45 4,25
270,94 15,65
153,11 14,62

B sung vin
Placebo
0,56 0,16
60,84 4,28
215,17 15,73
73,31 15,78

21,74 3,51
79,39 2,62
565 17,85
6,79 0,15

25,00 4,73
67,48 3,73
524 14,83
6,80 0,21

Bng XII.3.2.3.c. Ch tiu sinh hc tinh dch th sau 20 ngy b sung vin
Hasamine, Placebo
Ch tiu theo di
Lng tinh dch (ml)
Hot lc tin thng (A %) ca tinh trng
Nng tinh trng (triu/ml)
Tng s tinh trng tin thng (V.A.C:triu/
mt ln ly tinh)
T l k hnh (%) ca tinh trng
T l sng (%) ca tinh trng
Sc khng ca tinh trng i vi (NaCl 1%)
PH tinh dch

B sung vin
Hasamin
0,87 0,15
74,70 4,16
279,84 14,72
181,87 14,62

B sung vin
Placebo
0,63 0,17
62,73 3,25
225,84 11,84
89,25 15,78

18,80 3,66
80,72 3,69
572 14,84
6,81 0,12

24,72 3,96
69,83 4,74
528 15,61
6,83 0,15

Kt qu cho thy: b sung vin Vin Hasamin c tc dng lm tng c

ngha v phm cht tinh dch th so vi b sung Placebo.
Trc khi b sung ch phm, lng tinh dch th khc nhau khng c
ngha gia cc l th nghim. Sau 10 ngy b sung, lng tinh dch ca l 1:
0,78ml cao hn c ngha so vi lng tinh dch ca l 2: 0,56ml v trc khi b
sung. Sau 20 ngy b sung ch phm, lng tinh dch ca cc l cao hn khng
c ngha so vi sau 10 ngy. Lng tinh dch ca l1 cao hn c ngha ngha so
vi l 2.
Trc khi b sung ch phm, hot lc tin thng ca tinh trng khc nhau
khng c ngha gia cc l th nghim. Sau 10 ngy b sung ch phm, c tc
dng tt lm tng hot lc tin thng ca tinh trng l 1. Sau 20 ngy, hot lc
tin thng ca tinh trng l1: 74,70% cao hn c ngha so vi l2: 62,73%.
675

Hot lc tin thng ca tinh trng l 1 sau khi b sung ch phm cao hn c
ngha so vi trc khi b sung.
Nng tinh trng khc nhau khng c ngha trc khi b sung ch
phm. B sung ch phm c tc dng tng c ngha v nng tinh trng l 1
so vi l 2 v trc khi b sung.
B sung ch phm c tc dng tng tng s tinh trng tin thng. Sau 20
ngy b sung, tng s tinh trng c hot lc tin thng l 1:181,87 triu, cao
hn c ngha so vi l 2: 89,25 triu/ln.
T l k hnh ca tinh trng trc khi b sung ch phm khc nhau khng
c ngha. Sau 10 ngy b sung ch phm, t l k hnh ca tinh trng l 1:
21,74% thp hn c ngha thng k so vi l 2: 25,00%.
T l sng ca tinh trng khc nhau khng c ngha trc khi b sung ch
phm. Ch phm c nh hng tt lm tng t l sng ca tinh trng. Sau 20
ngy b sung Hasamin t l sng ca tinh trng l 1: 80,72% cao hn c
ngha so vi l 2: 69,83%.
Ch phm c nh hng tng sc khng ca tinh trng l 1 so vi trc b
sung, nhng khng c ngha.
Ch phm duy tr c pH n nh.
XII.4. Kt lun.
# Sn phm vin Hasamin phn tch cho thy hm lng qua thc nghim trn
ng vt cho thy an ton v c tc dng rt tt. Vin Hasamin l nhng
ngun thc phm chc nng c gi tr dinh dng. Th hin cc hm lng
axit amin, hoocmn steroit v cc nguyn t vi lng l cao so vi mt s loi
sn phm tng t khc trong nc v nc ngoi.
# Cc th nghim v c tnh cp v c tnh bn trng din trn chut nht
trng v th cho thy: khng nhn thy biu hin ng c no trn chut th
nghim trong thi gian theo di, Khng xc nh c liu gy cht 50%
ng vt th nghim (LD50) v khng tm c liu gy cht chut mc d
cho ung vi mc liu cao, khng nhn thy s khc nhau ng k v tnh
trng th, cc ch s huyt hc v sinh ho, i th cc tng ph sau th
nghim gia nhm chng v nhm th. Hnh nh tn thng m bnh hc t
chc gan v thn trn tiu bn vi th khng c s khc nhau nhiu gia nhm
chng v nhm th.
# Vin Hasamin c tc dng thc s r nt thng qua vic thc nghim tng
cht lng tinh dch trn th.

676

phn III. Kt lun v kin ngh chung

A. kt lun
ti KC 09.09/06-10 Nghin cu sng lc cc cht c hot tnh sinh hc
theo nh hng khng sinh, gy c t bo v chng xy ho t sinh vt bin
nhm to cc sn phm c gi tr dc dng l ti nghin cu lin ngnh, t

vn nghin cu mt cch c h thng ngun dc liu bin Vit Nam p dng

m hnh nghin cu ho hc theo nh hng hot tnh sinh hc (bioasay-guided
chemical study). Nh chng ti trnh by trn, y l m hnh nghin cu
tin tin nht hin nay trn th gii, c nhiu quc gia nh M, Nht Bn, Hn
Quc, Php p dng hnh thnh nhng chng trnh nghin cu ln qui m
quc gia v xuyn quc gia. Kt qu l trong nhng nm va qua, nhiu loi
thuc c hiu cha tr cc bnh him ngho c ra i v lu hnh trn th
gii thng qua cc chng trnh ny. Ngay cc nc trong khu vc nh Thi Lan,
Malaysia, Philippines cng c nhng chng trnh sng lc cc cht c hat
tnh sinh hc t bin v t c nhiu kt qu ban u ng khch l. Hiu
qu ca nhng chng trnh sng lc ny ph thuc rt nhiu vo i ng cn b
cc chuyn ngnh lin quan, h thng trang thit b phn tch v c bit l h
thng thit b th hot tnh sinh hc hin i, c tin cy cao. Mc thnh
cng cng ph thuc rt nhiu vo kinh ph dnh cho nghin cu.
trin khai thc hin ti ny, Ban Ch nhim ti KC09.09/06-10
tp trung c i ng cn b khoa hc ng o thuc cc lnh vc chuyn mn
khc nhau nh Ho, Sinh, Y, Dc, Cng ngh. Cc c quan tham gia ti u
l nhng c quan u ngnh ca Vit Nam cng tham gia gii quyt nhim v.
Sau 3 nm trin khai thc hin, vi n lc c gng ca tp th cn b thc hin
ti, c s quan tm, ng vin rt ln v kp thi ca Ban Ch nhim chng
trnh KC 09/06-10, B Khoa hc v Cng ngh, Vin Khoa hc v Cng ngh
Vit nam, ti hon thnh tt cc mc tiu ra ban u v t c
nhng kt qu tt. Cn c vo cc kt qu t c, c th rt ra cc kt lun
sau:
I. t chc 9 t kho st thu thp mu sinh vt bin trn cc vng bin bc,
trung v nam Vit Nam, thu thp v xc nh tn phn loi 101 mu sinh vt bin
v to tiu bn lu tr lu di tp trung vo cc nhm san h mm, da gai v hi
min. xy dng c c s d liu qun l cc loi sinh vt bin cha dc
liu di ven b Vit Nam. C s d liu cho php qun l, truy cp, b sung, tu
chnh thun tin v n gin.
II. tin hnh to dch chit tng v cc dch chit phn on ca 101 mu thu
c theo mt qui trnh thng nht phc v cho sng lc hot tnh sinh hc v
nghin cu ha hc.
III. tin hnh sng lc hot tnh sinh hc ca tt c cc dch chit tng v cc
dch chit phn on ca 101 mu sinh vt bin theo cc nh hng:

677

a. Hot tnh khng vi sinh vt kim nh (Antimicrobial activity asay)

Kt qu thu c cho thy, s mu c hot hot tnh khng vi sinh vt
kim nh l 51 trong tng s 101 mu sng lc (chim t l 50,49 %). Trong ,
s mu c hot tnh khng 1 vi sinh vt kim nh l 8 mu (chim 7,92 %),
khng 2 vi sinh kim nh l 10 mu, khng 3 vi sinh kim nh l 19 mu, khng
4 vi sinh kim nh l 10 mu, khng 5 vi sinh kim nh l 2 mu, khng 6 vi
sinh kim nh 1 mu v khng 7 vi sinh kim nh l 1 mu. Cc mu c hot
tnh c tin hnh chit phn b gia nc v cc dung mi hu c c
phn cc tng dn t hexan sang butanol. xc nh gi tr c ch ti thiu
(MIC) ca dch chit MeOH tng v cc phn on chit phn b.
b. Hot tnh gy c t bo trn 3 dng t bo ung th ngi (Cytotoxic activity
assay)
Trong s 101 mu th nghim, c 43 mu c biu hin hot tnh gy c
t bo (chim 42,57 %), trong c 12 mu khng mt dng t bo (chim 11,88
%), 15 mu khng 2 dng t bo (chim 14,85 %) v 16 mu khng c 3 dng t
bo (chim 15,84 %). xc nh gi tr IC50 (nng c ch 50% s pht trin
ca t bo) ca dch chit MeOH tng v cc phn on chit phn b nh
hng cho cc nghin cu tip theo.
c. Hot tnh chng oxy ho theo c ch bao vy gc t do DPPH (Antioxidant
activity assay)
Kt qu thu c cho thy, c 23 mu trong tng s 101 mu (chim t l
22,72 %) c biu hin dng tnh vi gi tr SC > 50%. Trong , c 18 mu c
c kh nng bao vy t 50-70% cc gc t do trong th nghim DPPH v 5 mu
c kh nng bao gy trn 70% cc gc t do. nh hng cho cc nghin cu
tip theo, gi tr SC50 (gi tr nng mu th c kh nng bao vy 50% cc gc
t do) ca dch chit MeOH tng v cc phn on chit phn b ca cc mu c
biu hin hot tnh c xc nh.
IV. Trn c s cc kt qu nghin cu thu thp, gim nh loi, to dch chit v
sng lc hot tnh sinh hc, kt hp vi cc tiu ch la chn b sung nh: Mc
kh khn trong cng tc ly mu, tr lng thc t ca loi v kh nng c th
thu thp mu, tra cu cc nghin cu v hot tnh sinh hc v thnh phn ha hc
cng b v loi hoc chi ca i tng nghin cu..., la chn c 07 loi
sinh vt bin thuc cc nhm san h mm, da gai v hi min thu mu lng
ln v nghin cu su v mt ho hc l :
- Nhm san h mm :
1. T loi san h mm Sarcophyton mililatensis, chit xut, phn lp v xc
nh cu trc c 10 hp cht trong c 07 hp cht ditecpen dng
cembranoit (05 cht mi), 01 hp cht 9,11-secosterol mi (sarcomilasterol) v
02 hp cht sterol mang nhiu nhm hydroxyl. tin hnh nh gi hot tnh
gy c t bo, chng xy ha v chng long xng ca cc hp cht phn lp
c. Cc kt qu c ng ti trn 02 bi bo chuyn ngnh quc gia v
02 bi bo chuyn ngnh quc t.
2. T loi san h mm Cladiella sp., 08 hp cht c phn lp v xc nh
cu trc, trong c: 02 hp cht 9,11-secosterol (01 cht mi l :
678

cladiasterol), 01 hp cht ceramit mi (cladiaceramide) v 01 hp cht

cerebrosit mi (cladiacerebroside). tin hnh nh gi hot tnh gy c t
bo, chng xy ha v khng NF-B ca cc hp cht phn lp c. ng
ti kt qu trn 01 tp ch chuyn ngnh quc t.
- Nhm da gai:
3. T loi hi sm Holothuria scabra, tin hnh chit xut, phn lp v xc
nh c cu trc ca 09 hp cht, trong c 06 hp cht tritecpen saponin
dng khung holosthan (03 hp cht mi l: holothurin A3, holothurin A4 v
holothrigenin B). tin hnh nh gi hot tnh gy c t bo v khng sinh
ca cc hp cht. Tt c cc hp cht saponin u th hin hot tnh gy c t
bo cao. Cc hp cht holothurin A2 v A3 c nh gi hot tnh gy c
t bo bng phng php quan st trc tip hnh thi b mt t bo di knh
hin vi sau 24, 48 v 72 gi. Cc kt qu c ng trn 02 tp ch chuyn
ngnh quc t v 01 hi ngh quc gia. Ngoi ra, tp th tc gi cng ng
k v c cp 01 bng c quyn sng ch s 6852 ti Cc S hu Tr tu
Vit Nam v phng php phn lp v hot tnh chng ung th ca hp cht
mi holothurin A3. Cc kt qu nghin cu c gi tham gia gii thng
khoa hc k thut Trung ng on Thanh nin ln th VIII - 2008 v ot
gii khoa hc: s 1890/GTKHKTTN-2008.
4. T loi sao bin Archaster typicus, 07 hp cht c phn lp v xc nh
cu trc, trong c 01 hp cht cerebrosit mi (archastercerebroside) v 03
hp cht sterol mang nhiu nhm hydroxy (highly oxygenated sterols). Hot
tnh gy c t bo v khng NF-B ca cc hp cht c nh gi.
ng c 02 bi bo trn tp ch quc gia, 01 bi trn Hi tho chuyn ngnh
quc gia v 01 bi trn Hi tho chuyn ngnh quc t.
- Nhm hi min:
5. T loi hi min Xestospongia testudinaria, 10 hp cht c phn lp v
xc nh cu trc, trong ng lu nht l c 02 hp cht axit bo khng no
b brm ha c cu trc mi l: axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19triene-7,9,17-triynoic v 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19triynoic. tin hnh nh gi hot tnh khng vi sinh vt kim nh v gy
c t bo ca cc hp cht. Hai hp cht mi u khng mnh ba chng vi
sinh vt kim nh l: trc khun m xanh Pseudomonas aeruginosa, khng t
cu vng Staphylococcus aureus v khng nm si Fusarium oxysporum vi
gi tr IC50 thp l 50 g/ml. Tp th tc gi ng c 02 bi bo trn tp
ch chuyn ngnh quc gia. Ngoi ra, tp th tc gi gi bn tho v n
ng k 01 bng c quyn sng ch (s n 1-2007-01616 SC ngy 8/8/2008)
ti Cc S hu Tr tu Vit Nam v hp cht axit 20-bromo-(11E,15E,19E)eicosa-11,15,19-triene-7,9,17-triynoic v ang c tin hnh thm nh ni
dung khoa hc.
6. T loi hi min cnh xanh Gellius varius, phn lp v xc nh cu trc
c 10 hp cht. Trong c 02 hp cht sulfuno glycolipit c cu trc c
th vi s c mt ca d t lu hunh trong phn t. tin hnh nh gi hot
tnh gy c t bo v khng vi sinh vt kim nh ca cc hp cht. Cc kt
qu c ng ti trong 02 bi bo trn cc tp ch chuyn ngnh quc gia.

679

7. T loi hi min Ianthella sp., 05 hp cht steroit c cu trc vng propan

mch nhnh c phn lp v xc nh cu trc. Trong , 03 hp cht c
cu trc mi v c chng ti t tn l iansterol A, B v C. Kt qu nghin
cu hot tnh gy c t bo cho thy, tt c cc hp cht phn lp c u
th hin hot tnh cao trn 03 dng t bo ung th ngi c th nghim l
MCF-7 (v), SK-Hep-1 (gan) v Hela (c t cung).
V. nghin cu thm d kh nng khng vim t rong, to bin Vit Nam v
thu c cc kt qu sau:
- thu thp v nh tn c 19 loi rong bin Vit Nam phn b cc tnh
min Bc, Trung v Nam ca Vit Nam.
- xy dng c quy trnh tch dch chit c hot tnh khng vim t cc loi
rong bin Vit Nam bng dung mi metanol; phng php sng lc nhanh cc
loi rong to bin trong khun kh ti c hot tnh khng vim bng dung
mi metanol.
- Trn c s ca phng php ny, trong tng s 19 loi rong bin c sng lc
th Sargassum swartzii v Ulva reticulata c hot tnh khng vim cao v c
tim nng khai thc c pht hin.
- tin hnh th hot tnh khng vim v khng sinh ca cc dch chit thu
c t Sargassum swartzii v Ulva reticulata.
- xc nh c hm lng lipit tng s v thnh phn axit bo ca hai mu
cao rong Sargassum swartzii v Ulva reticulata
- phn lp v xc nh cu trc c hp cht 3,5-dimethyl-4-al-pyrol t cao
diclometan v hp cht 22-dehydroclerosterol t cao n-hexan ca rong rong
Sargassum swartzii.
VI. nghin cu thm d kh nng khng sinh t vi sinh vt bin v thu c
cc kt qu sau:
- phn lp v lm sch c 1052 chng vi sinh vt bin t 81 mu nc bin,
trm tch bin v ng thc vt bin Vit Nam. Trong , c 552 chng vi
khun, 253 chng nm si, 157 chng x khun v 90 chng nm men.
- sng lc s b v th kh nng khng 5 chng VSV kim nh ca 1052
chng vi sinh vt bin.
- th kh nng dit su hi, gy c t bo ca 142 chng vi sinh vt bin.
- xc nh kh nng sinh cc enzim ngoi bo amylaza, xenlulaza v proteaza
ca 82 chng vi sinh vt bin
- xc nh hm lng axit bo khng bo ho mch di ca 6 chng nm men
bin.
- tuyn chn c 11 chng vi sinh vt bin c hot tnh sinh hc cao v nh
tn khoa hc 08 chng.
- a ra qui trnh nui cy v tch chit cht khng khun t dch nui cy
chng nm si T10.6
VII. nghin cu hon thin quy trnh cng ngh v trin khai sn xut c
04 sn phm thc phm chc nng l: thc phm chc nng CEFISH b sung
omega-3 ci thin chc nng chuyn ha lipit mu, huyt p bnh nhn tim

680

mch, vin tng lc BIONAMINE tng cng th lc cho vn ng vin t sao

bin, vin nang mm HALIOTIS t bo ng c tc dng bi b c th v thc
phm chc nng vin nang mm HASAMIN t hi sm.
VIII. nghin cu c tnh cp v c tnh bn trng din ca cc ch phm
trn ng vt thc nghim. Kt qu cho thy khng sn phm no th hin c c
tnh cp v c tnh bn trng din trn ng vt th nghim. tin hnh cc
nghin cu dc l cc sn phm thc phm chc nng v thu c cc kt qu
tt.
IX. xy dng c tiu chun c s cht lng sn phm v c cp giy
chng nhn tiu chun sn phm ca B Y t, cho php lu hnh trn ton quc
ca tt c bn sn phm: sn phm CEFISH c giy chng nhn tiu chun sn
phm s 6585/2007/YT/CNTC, sn phm BIONAMINE c giy chng nhn
tiu chun sn phm s 956/2008/YT-CNTC, sn phm HALIOTIS c cp giy
chng nhn tiu chun sn phm s 6714/2007/YT-CNTC v sn phm
HASAMIN c cp giy chng nhn tiu chun sn phm s 4978/2006/YTCNTC.
X. sn xut lng ca tt c 04 sn phm phc v cho cc nghin cu
v c tnh v tc dng dc l theo h s ng k.
XI. Cc kt qu khc: ng k 02 c quyn sng ch, xut bn c 01 sch
chuyn kho, ng 17 bi bo trong c 11 bi trn cc tp ch chuyn ngnh
quc gia v 06 bi quc t (05 bi ng v 01 bi gi ng), ng 09 bo
co ti cc hi ngh trong c 04 hi ngh quc t v 05 hi ngh trong nc,
o to 02 thc s bo v thnh cng lun n v 02 tin s (01 tin s bo v
thnh cng lun n v 01 ang tin hnh).

b. kin ngh
Da trn cc kt qu thu c, tp th cn b thc hin ti xut mt
s kin ngh sau:
1. Cc nghin cu tip theo cn tip tc tp trung vo cc nhm i tng c tim
nng dc dng cao l san h mm, da gai v hi min vi tng hot tnh c
th nh : Gy c t bo, chng long xng, khng sinh
2. Cn m rng quy m sn xut ca cc sn phm phn phi trn c nc,
phc v cho cng tc chm sc sc khe v nng cao cht lng cuc sng ca
ngi dn.
3. Tip tc trin khai nghin cu to sn phm t cc ngun nguyn liu di do
v c nhiu trin vng khc nhm to ra cc sn phm c gi tr tng t gp
phn nng cao gi tr ngun dc liu bin, ng thi c cc bin php bo tn
v khai thc hp l ngun ti nguyn v gi ny.
4. Tip tc pht trin nghin cu su hn na, tm kim cc thnh phn c hot
tnh chnh trong cc nguyn liu nghin cu, pht trin thnh cc ch phm
thuc cha bnh phc v cuc sng.

681

Li cm n
Tp th cn b thc hin ti xin by t lng bit n n
- Ban ch nhim chng trnh KH&CN trng im cp nh nc KC 09/0610: Khoa hc v Cng ngh Bin phc v pht trin bn vng kinh t x hi
- Vn phng cc chng trnh, B Khoa hc v Cng ngh Vit Nam
- Vin Ha hc cc Hp cht Thin Nhin.
ng vin v gip chng ti trong qu trnh thc hin ti.
Nhn dp ny chng ti cng xin chn thnh cm n cc n v cng
tham gia tch cc vi Vin Ho hc cc hp cht thin nhin, cc nh khoa hc
trong v ngoi Vin cng cng tc ti c hon thnh ng theo k
hoch.

682

cc cng trnh cng b trong khun kh ti

1. Danh mc cc sng ch
TT Tn tc gi
1 Chu Vn Minh, Phan
Vn Kim, Nguyn Hi
ng, Nguyn Vn
Thanh, L Mai Hng,
Cng Thung, Trn
Thu Hng, Nguyn
Xun Cng.
2 Chu
Vn
Minh,
Nguyn Xun Cng,
Phan Vn Kim, L Mai
Hng, Cng Thung

2. Danh mc cc sch
TT Tn tc gi
1 Chu Vn Minh, Phan
Vn Kim, Nguyn Hi
ng

Tn sng ch
Hp cht Holothurin A3 c hot tnh
chng ung th v phng php chit hp
cht ny t loi hi sm Holothuria
scabra, Bng c quyn sng ch s
6852, cp theo quyt nh s 2394/QSHTT, ngy 14 thng 02 nm 2008 ca
Cc S hu Tr Tu Vit Nam
Hp cht axit 20-bromo-(11E,15E,19E)eicosa-11,15,19-triene-7,9,17-triynoic v
phng php chit hp cht ny t loi hi
min Xestospongia testudinaria, s n 12007-01616 SC ngy 8/8/2007

Ghi ch
c
cp

chp
nhn
n

Tn sch, nh xut bn
Ghi ch
Cc hp cht c hot tnh sinh hc t sinh xut
vt bin. Nh xut bn Khoa hc K bn
thut, 2007, 241 trang.

3. Danh mc cc tp ch quc t
TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Nguyen Xuan Cuong, New Cembranoid Diterpenes from the
Tran Anh Tuan, Phan Vietnamese Soft Coral Sarcophyton
Van Kiem, Chau Van mililatensis
Stimulate
Osteoblastic
Minh, Eun Mi Choi, and Differentiation in MC3T3-E1 Cells.
Young Ho Kim.
Chemical & Pharmaceutical Bulletin,
2008, 56(7), 988-992.
2 Chau
Van
Minh, A new 9,11-secosterol from the
Nguyen Xuan Cuong, Vietnamese Sea soft coral Sarcophyton
Tran Anh Tuan, Eun Mi mililatensis, increases the function of
Choi, Young Ho Kim, osteoblastic MC3T3-E1 cells, Natural
and Phan Van Kiem.
Product Comunications (USA), 2007,
2(11), 1095-1100.
3 Nguyen Hai Dang, Two new triterpene glycosides from the
Nguyen Van Thanh, Vietnamese sea cucumber Holothuria
Phan Van Kiem Le Mai scabra, Archives of Pharmacal Research,
Huong, Chau Van Minh, 2007, 30(11), 1387-1391.
and Young Ho Kim.
4 Chau Van Minh, Phan New 9,11-secosterol glycoside and
Van Kiem, and Nguyen ceramide from a soft coral Cladiella sp.
Xuan Cuong.
living in Vietnamese sea, ASEAN Journal
on Sicience and Technology for
Development, 2007, 24(3), 275-283.
5 Nguyen Van Thanh, A new triterpene glycoside from the sea
Nguyen Hai Dang, Phan cucumber Holothuria scabra collected in
683

Ghi ch
ng

ng

ng

ng

ng

Van Kiem, Nguyen

Xuan Cuong, Hoang
Thanh Huong, and Chau
Van Minh.
Chau
Van
Minh,
Nguyen Huu Tung,
Nguyen Xuan Nhiem,
Phan Van Kiem, Tran
Thu Ha, Jae-Hee Hyun,
Hee-Kyoung
Kang,
Young Ho Kim

Vietnam. ASEAN Journal on Science and

Technology for Development, 2006,
23(4), 253-259.
C29-Sterols with a cyclopropane ring at C- gi
25 and 27 from the Vietnamese marine ng
sponge Ianthella sp.

4. Danh mc cc tp ch quc gia

TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Chau
Van
Minh, Highly oxygenated sterols from the
Nguyen Xuan Cuong, starfish Archaster typicus. Journal of
Nguyen Van Thanh, Chemistry (Vietnam), 2007, 45(3), 377Hoang Thanh Huong, 381.
and Phan Van Kiem.
cholesterol
and
2 Phan
Van
Kiem, Cholest-7-en-3-one,
Nguyen Van Thanh, batilol from the starfish Archaster typicus.
Nguyen Xuan Cuong, Journal of Chemistry (Vietnam), 2007,
and Chau Van Minh.
45(4), 513-517.
3 Nguyn Xun Cng, Cc hp cht glycolipit t loi hi min
Phan Vn Kim, Chu cnh xanh Gellius varius sinh sng ti
Vn Minh.
vng bin Vit Nam, Tp ch Khoa hc v
Cng ngh, 2007, 45 (2), 41-47.
4 Nguyen Xuan Cuong, Sarcophytol X, a new cembranoid
Tran Anh Tuan, Phan diterpene from the vietnamese sea soft
Van Kiem, and Chau coral Sarcophyton mililatensis, Tp ch
Van Minh.
khoa hc (i hc S phm H Ni II),
2007, 1, 55-60.
5 Chau
Van
Minh, Polyhydroxylated sterols from the
Nguyen Xuan Cuong, Vietnamese Sea soft coral Sarcophyton
Tran Anh Tuan, Eun Mi mililatensis, increases the function of
Choi, Young Ho Kim, osteoblastic MC3T3-E1 cells, Tp ch
and Phan Van Kiem.
khoa hc (i hc S phm H Ni II),
2007, 1, 108-116.
6 Nguyn Xun Cng, Nghin cu thnh phn ha hc loi hi
Trn Anh Tun, Nguyn min Xestospongia testudinaria thu thp
Thu Phng, V Vn ti Vit Nam. Tp ch Khoa hc v Cng
Thnh, Phan Vn Kim, ngh, 2007, 45(3), 43-50.
Chu Vn Minh.
7 Nguyen Xuan Cuong, Isoprenoids from the sponge Gellius
Nguyen Hai Dang, Do varius living in Vietnamese sea, Journal
Cong Thung, Tran Thu of Chemistry (Vietnam), 2007, 45(6), 776Huong, Nguyen Thi 780.
Kim Thuy, Phan Van
Kiem, Chau Van Minh.

684

Ghi ch
ng

ng

ng

ng

ng

ng

ng

10

11

Chau
Van
Minh,
Nguyen Hai Dang,
Nguyen Xuan Cuong,
Phan Van Kiem, Hoang
Thanh Huong,
Nguyn Minh Thanh,
Luyn Quc Hi, Li
Kim Dung, Phan Vn
Kim, Chu Vn Minh,
ng Dim Hng
ng
Dim
Hng,
Hong Minh Hin,
Nguyn Minh Thanh,
Chu
Vn
Minh,
Nguyn Trng Thng
Chau Van Minh, Phan
Van Kiem, Tran Anh
Tuan, and Nguyen Xuan
Cuong.

Bioacive constituents from marine ng

organisms inhabiting in Vietnames sea.
Journal of Science and Technology
(Vietnam), 2007, 45(6-DB), 1-18.
Xc nh bn cht ha hc ca cht khng ng
vim c trong dch chit ton phn t
Sargassum swartzii ca Vit Nam, Tp
ch Cng ngh Sinh hc, 2008, S 6(4A),
713-720.
Nghin cu kh nng khng vim t rong ng
to bin Vit Nam, Tp ch Ha hc,
2008, Tp 46, s 53, trang 81-90.
Two new brominated fatty acids from the ng
sponge Xestospongia testudinaria living in
Vietnamese sea. Advances in Natural
Sciences, 2008, 9(2), 167-171

5. Danh mc cc hi ngh quc t

TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Chau
Van
Minh, A new galactocereboside from the starfish
Nguyen Van Thanh, Archaster typicus, 12t Asian Chemical
Nguyen Xuan Cuong, Congress (12ACC), August 23-25, 2007,
Phan
Van
Kiem, Malaysia, p. 309.
Nunziatina De Tommasi
2 Chau
Van
Minh, Two new triterpene glycosides from the
Nguyen Van Thanh, Vietnamese sea cucumber Holothuria
Nguyen Hai Dang, Phan scabra, 12t Asian Chemical Congress
Van Kiem, Le Mai (12ACC), August 23-25, 2007, Malaysia,
p. 309.
Huong, Young Ho Kim
3 Phan Van Kiem, Chau Bioactive Constituents from Marine
Van Minh, Nguyen Hai organisms inhabiting in vietnamese sea,
Dang, Nguyen Xuan Oral prsentation in Intenational
Cuong, Hoang thanh Conference From the new drug
Development to Human Well-being The
huong, Kim Young Ho
Pharmaceutical Society of Korea, April
30 - May3, 2008, Vol. 1A, p.15.
4 Nguyen Minh Thanh, The screening compound of antiHoang Minh Hien, inflamatory activities from Vietnamese
Hoang Sy Nam, Chau seaweeds. Vth Asian Pacific Phycological
Van Minh, Dang Diem Forum, Wellington, New Zealand,
November 10-14, 2008. p.88
Hong.
6. Danh mc cc hi ngh quc gia
TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Nguyen Van Thanh, Steroidal constituents from Holothuria
Chau Van Minh, scabra living in Vietnamese sea. Tuyn tp

685

Ghi ch
ng

ng

ng

ng

Ghi ch
ng

Hoang Thanh Huong, cc bi bo khoa hc Hi ngh Khoa hc ln

Phan Van Kiem.
th 20-HBK H Ni, phn ban Cng ngh
Ha hu c, 2006, 200-204.
2

Chu
Vn
Minh, Mt galactocereboside mi t sao bin
Nguyn Vn Thanh, Archaster typicus. Hi ngh Ha hc hu c
Nguyn Xun Cng, Ton quc ln th 4, H Ni, thng 10 nm
Phan
Vn
Kim, 2007, trang 446-450.

ng

Bc u nghin cu sng lc cht c hot

tnh khng vim t mt s loi rong bin
Vit Nam, Nhng Vn nghin cu c bn
trong khoa hc s sng, 2007, 773-776.

ng

Nghin cu tuyn chn vi sinh vt bin c

kh nng i khng VSV kim nh, gy
cht cn trng v kh nng ng dn trong
nng nghip v y dc. Hi ngh Khoa hc Cc bin php phng tr su bnh khng
gy nhim mi sinh, 2007, 77-83.

ng

Nunziatina
De
Tommasi, Lm Ngc
Thim
Hong S Nam, inh
Khnh Chi, Hong
Minh Hin, Chu
Vn Minh, ng
Dim Hng
Tng Kim Thun,
ng Th Mai Anh,
L Th Thanh Xun,
Phm Cng Hot,
Chu Vn Minh

Tng Kim Thun, L

Th Thanh Xun,
ng Th Mai Anh,
Phm Cng Hot,
Chu Vn Minh

Nghin cu sinh tng hp v tch chit cht ng

khng khun t chng nm si Penicillium
waksmani Zaleski T10.6, phn lp t trm
tch bin Bnh nh. Hi ngh Khoa hc ton
quc ln th IV - Ha sinh v sinh hc phn
t phc v nng, sinh, y hc v cng nghip
thc phm, 2008, 892-896.

7. Danh mc cc lun n
TT Tn hc vin
Tn lun n
1 Nguyn Th Kim Nghin cu thnh phn ha hc v hot tnh gy
Thy
c t bo ca loi hi min Gellius varius. Lun
vn thc s Ha hc, 2007.
2 Trn Anh Tun
Nghin cu thnh phn ha hc v hot tnh sinh
hc ca loi san h mm Sarcophyton
mililatensis, Lun vn thc s Ha hc, 2008.
3 Nguyn
Vn Nghin cu thnh phn ho hc v kho st hot
Thanh
tnh gy c t bo in vitro ca loi hi sm
Holothuria scabra v sao bin Archaster typicus
sinh sng ti vng bin vit nam, Lun n Tin
s Ho hc, 2008
4 Trn Thu H
Nghin cu thnh phn ha hc v hot tnh sinh
hc mt s loi hi min sinh sng ti vng bin
Vit Nam, Lun n Tin s Ha hc, 2008-2012.

Ghi ch
bo
v
bo
v
bo
v

ang
thc
hin

Ngoi ra ti t chc 01 hi tho:

-Hi tho khoa hc ti KC09.09/06-10 thng 12 nm 2008 ti H Ni

686

Ti liu tham kho

1
2
3
4
5
6
7
8
9
10
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Nguyn Tp v Trn Ton. Cy thuc v ng vt lm thuc Vit Nam, tp II, Nh xut
bn Khoa hc v K thut H Ni, 2004.

698

B khoa hc v cng ngh

chng trnh KH&CN Bin phc v pht trin bn vng
kinh t-x hi, m s kc.09/06-10

Bo co tm tt ti
Nghin cu sng lc cc cht c hot tnh sinh
hc theo nh hng khng sinh, gy c t bo
v chng xy ho t sinh vt bin nhm to cc
sn phm c gi tr dc dng
M s: KC.09.09/06-10
Ch nhim ti: GS.TS Chu Vn Minh
C quan ch tr ti: Vin Ho hc cc Hp cht thin nhin
Vin Khoa hc v Cng ngh Vit Nam

H Ni, 5 nm 2009

B khoa hc v cng ngh

chng trnh KH&CN Bin phc v pht trin bn vng
kinh t-x hi, m s kc.09/06-10

Bo co tm tt ti
Nghin cu sng lc cc cht c hot tnh sinh
hc theo nh hng khng sinh, gy c t bo
v chng xy ho t sinh vt bin nhm to cc
sn phm c gi tr dc dng
M s: KC.09.09/06-10
C quan ch tr ti: Vin Ho hc cc Hp cht thin nhin
Vin Khoa hc v Cng ngh Vit Nam
Chu Vn Minh - Ch nhim ti
Phan Vn Kim - Th k ti, L Mai Hng, Phm Quc
Long, Hong Thanh Hng, Nguyn Minh H, Cng
Thung, Tng Kim Thun, Nguyn Huy Nam, Trng
Hng Lan, ng Dim Hng, on Thi Hng.

Thng tin tm tt v ti
1.Tn v m s ti:
Nghin cu sng lc cc cht c hot tnh sinh hc theo nh hng khng sinh,
gy c t bo v chng xy ho t sinh vt bin nhm to cc sn phm c gi tr
dc dng
Thuc chng trnh KH&CN trng im cp nh nc KC09/06-10: Khoa hc v Cng
ngh Bin phc v pht trin bn vng kinh t x hi
M s: KC09.09/06-10
Thi gian thc hin: 1/2006-12/2008.
2. C quan ch tr ti:

Vin Ho hc cc Hp cht thin nhin

18 Hong Quc Vit, Cu giy, H Ni
in thoi: 04-8360830; Fax: 04-7564390
C quan phi hp chnh: Vin Ti nguyn v Mi trng bin Hi Phng, Vin Y
hc c truyn Qun i, Vin Cng ngh Sinh hc, Vin
Cng nghip Thc phm, Cng ty Dc phm ng
Dng
3. Ch nhim ti:
GS. TS. Chu Vn Minh
a ch: Vin Ho hc cc Hp cht thin nhin
Vin Khoa hc v Cng ngh Vit Nam
in thoi: 04-8363375; Fax: 04- 7564390
Email: cvminh@vast.ac.vn
Th k khoa hc:
TS. Phan Vn Kim
a ch: Vin Ho hc cc Hp cht Thin nhin
Vin Khoa hc v Cng ngh Vit Nam
in thoi: 04-7562378; Fax: 04- 7564390
Email: phankiem@vast.ac.vn
4. Mc tiu ca ti:
4.1. Xy dng c danh mc sinh vt bin c cht khng sinh, gy c t bo v
chng xy ho.
4.2. Xy dng c qui trnh cng ngh tch chit cc cht c hot tnh sinh hc
v to ra mt s sn phm c gi tr dc dng.
4.3. o to cn b trong lnh vc Hp cht thin nhin bin, mt lnh vc cn rt
mi m Vit nam.
4.4. Phi hp vi cc c quan lin quan a vo ng dng thc t.
5. Nhng ni dung chnh ca ti:
5.1. Thu thp mu, phn loi sinh vt bin, xc nh tn phn loi, to tiu bn, xy
dng c s d liu v ngun dc liu bin Vit Nam.
5.2. Sng lc hot tnh sinh hc cc mu sinh vt bin theo nh hng khng sinh,
gy c t bo v chng xy ho.
5.3. Nghin cu thnh phn ho hc v hot tnh sinh hc ca mt s loi sinh vt
bin c la chn thng qua qu trnh sng lc.
5.4. Nghin cu thm d kh nng khng vim t rong - to v kh nng sinh cc cht
c hot tnh sinh hc t vi sinh vt bin
5.5. Xy dng quy trnh cng ngh to ch phm, xy dng v thm nh tiu chun
c s, tin hnh cc nghin cu v dc l v ng k tiu chun sn phm v
giy php lu hnh ca B Y t cho 04 ch phm: CEFISH, BIONAMINE,
HALIOTIS v HASAMIN
1

Danh sch nhng ngi thc hin

A. Tc gi chnh
1. GS TS Chu Vn Minh
2. TS Phan Vn Kim
3. PGS TS L Mai Hng
4. PGS TS Phm Quc Long
5. PGS TS Hong Thanh Hng
6. TS Nguyn Minh H
7. TS Cng Thung
8. PGS TS Tng Kim Thun
9. ThS Nguyn Huy Nam
10. TS Trng Hng Lan
11. TS ng Dim Hng
12. ThS on Thi Hng

Ch nhim ti
Th k ti
Vin Ho hc cc Hp cht thin nhin
Vin Ho hc cc Hp cht thin nhin
Vin Ho hc cc Hp cht thin nhin
Vin Y hc C truyn qun i
Vin Ti nguyn v Mi trng Bin
Vin Cng ngh Sinh hc
Vin Cng ngh Sinh hc
Vin Cng nghip thc phm
Vin Cng ngh Sinh hc
Cng ty Dc ng Dng

B. Nhng ngi tham gia

1. TS Lu Vn Chnh
2. TS Nguyn Tin t
3. NCS Nguyn Hoi Nam
4. NCS on Lan Phng
5. NCS Trn Th Nh Hng
6. ThS Trn Hng Quang
7. ThS Trnh Thu Hng

Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN

27. BS Phm Th Dung

28. TS L Vn Ty
29. CN V Th Ninh
30. PGS TS V Mnh Hng
31. GS TS L Qu Phng
32. CN ng Trn Hon
33. BS Ng c Nhun

8. TS L Minh H

Vin HCTN

34. DS Nguyn Vn Chung

9. ThS Nguyn Xun Cng

10. CN Nguyn Phng Tho
11. CN Trn Hng Hnh
12. NCS Nguyn Hng Vn

Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN

13. ThS Nguyn Hi ng

Vin HCTN

35. TS Nguyn Vn X
36. ThS. Hong T Minh Hin
37. ThS. Hong Th Lan Anh
38. CN Hong S Nam
39. CN Nguyn T Minh
Thanh

14. ThS Hu Ngh

Vin HCTN

15. NCS Nguyn Vn Thanh

Vin HCTN

16. ThS Mai Ngc Ton

17. NCS Phm Hi Yn
18. CN Phan T Thanh Hng
19. CN Trn Anh Tun
20. NCS Trn Thu H
21. ThS. Nguyn T Kim Thy
22. Ths Li Quc Phong
23. KS Dng Vn ng
24. ThS. Trn T Minh H
25. CN Phm Linh Khoa
26. KS Nguyn Th Ln

Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin HCTN
Vin CNTP
Vin CNTP
Vin CNTP
Vin CNTP
Vin CNTP

40. CN Hunh Quang Nng

41. PGS TS Nguyn Trng
Thng
42. ThS Nguyn Th Vn Anh
43. TS Nguyn Cng Thc
44. ThS Nguyn Vn Lnh
45. NCS. Nguyn ng Ngi
46. NCS. Trn Mnh H
47. CN Nguyn Th Hong
48. CN L Th Thy
49. NCS. Nguyn Vn Qun
50. CN V Th Lu
51. TS. m c Tin
52. ThS. Chu Th Cng
2

BVK H Ty
Vin CNSH
Vin CNSH
Hc vin QY
Vin KHTDTT
Vin KHTDTT
Vin KHTDTT
Cng ty
INTECPHARM
TT GD&PT Sc k
Vin CNSH
Vin CNSH
Vin CNSH
Vin CNSH
Vin NC&UD CN
Nha Trang
i hc Y HN
i hc Y HN
Vin YHCT
Vin YHCT
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin
Vin TN&MT Bin

Tm tt bo co
B cc ca bo co c trnh by nh sau:
M u
Phn I. Tng quan, m hnh v phng php nghin cu
I. Tng quan v dc liu bin
II. M hnh nghin cu v phng php nghin cu.
Phn II. Kt qu nghin cu
A. Thu thp mu v sng lc hot tnh sinh hc
Chng I. Thu thp, nh loi mu SVB v xy dng c s d liu
Chng II. X l mu, to dch chit v sng lc hot tnh sinh hc
B. Nghin cu ha hc theo nh hng hot tnh sinh hc
Chng III. Nghin cu ho hc theo nh hng hot tnh sinh hc ca mt s
loi san h mm
Chng IV. Nghin cu ho hc theo nh hng hot tnh sinh hc ca mt s
loi da gai
Chng V. Nghin cu ho hc theo nh hng hot tnh sinh hc ca mt s
loi hi min
Chng VI. Tng hp cc cht phn lp c trong khun kh ti
C. Cc nghin cu v rong, to v vi sinh vt bin
Chng VII. Nghin cu thm d kh nng khng vim t rong v to bin Vit
Nam
Chng VIII. Nghin cu thm d kh nng sinh cc cht c HTSH t vi sinh vt
bin
D. Cc nghin cu ng dng sn xut cc sn phm
Chng IX. Nghin cu cng ngh sn xut thc n chc nng b sung Omega 3
phng v h tr iu tr bnh tim mch.
Chng X. Nghin cu quy trnh sn xut thc phm chc nng tng cng th lc
cho vn ng vin t sao bin
Chng XI. Nghin cu bo ch v nh gi tc dng bi b c th ca vin nang
HALIOTIS t bo ng
Chng XII. Nghin cu cng ngh sn xut vin nang mm hi sm HASAMIN
Phn III. Kt lun v kin ngh chung

Ni dung chnh ca ton b bo co c tm tt nh sau:

Phn I. Tng quan, m hnh v phng php nghin cu

I. Tng quan v dc liu bin
Phn tng quan c trnh by trong bo co gm cc mc sau:
I.1. Tng quan chung tnh hnh nghin cu trn th gii
1. Lc s hnh thnh, pht trin v cc hng nghin cu hot cht sinh hc bin
hin nay.
Mc ny trnh by mt cch tm tt s hnh thnh v pht trin ca nghin cu
dc liu bin, ng thi nu ra mt s hng nghin cu hot cht sinh hc bin hin
nay trn th gii. Trong , hng sng lc theo nh hng hot tnh sinh hc c
nh gi l mt bc tin trong vic nghin cu, pht trin cc dc phm mi v
ang c nhiu trung tm trn th gii s dng nh l mt trong nhng cng c hng
u pht hin ra cc dc t mi.
2. Cc hp cht thin nhin bin v hot tnh sinh hc t cc loi hi min, da gai v
san h mm.
Mc ny trnh by mt cch tng qut v cc hot cht c phn lp t hi min,
da gai v san h mm c hot tnh c bit hp dn hoc ang c tin hnh th
nghim tin lm sng v lm sng theo cc hng: khng vim, chng ung th, khng vi
rt, khng sinh, khng nm... in hnh nh: Contignasterol c phn lp t hi min
Petrosia congignata th hin hot tnh khng vim rt cao hot ng theo c ch km
hm s gii phng histamine t cc bch cu, cc dn xut tng hp ca n ang c
tin hnh nghin cu lm sng pha II iu tr bnh hen. Cc hp cht sarcodictyin phn
lp t loi san h mm Sarcodictyon roseum hin ang l nhng ng vin nng k cho
nghin cu to thuc tr ung th. Hng ngn hp cht khc v ang c nghin cu,
pht trin cc Vin nghin cu v Bnh vin trn ton th gii.
3. Kh nng khai thc ngun dc liu bin, c hi v thch thc
Trin vng v thch thc trong vic nghin cu cc hot cht t sinh vt bin c
trnh by vn tt trong mc ny. Vi u th v cu trc ha hc c o, a dng v th
hin nhiu hot tnh qu bu, cc hp cht thin nhin bin c nhiu tim nng
nghin cu v pht trin cc dc phm mi. Tuy nhin cng c khng t tr ngi nh:
s kh khn trong cng tc ly mu, kh nng thu mu lng ln.... Cc hot cht t
sinh vt bin thng c hm lng thp do thu c lng cht sn xut
thuc s cn mt lng nguyn liu khng l, v d thu c 1 gam ET-743 th phi
cn ti 1 tn E. turbunata.
4. Phng hng nghin cu sng lc hot tnh t ngun dc liu bin trn th gii
Mc ny a ra cc phng hng nghin cu sng lc hot tnh t ngun dc
liu ang c s dng trn th gii
I.2. Tng quan tnh hnh nghin cu trong nc
Mc ny trnh by mt cch tm tt v h thng cc nghin cu trong nc v cc
cht c hot tnh sinh hc t ngun dc liu bin nh: sng lc hot tnh sinh hc,
nghin cu ha hc theo nh hng HTSH v nghin cu ch to sn phm.
II. M hnh nghin cu v phng php nghin cu

II.1. La chn m hnh nghin cu v thit k nghin cu

II.1.1. La chn m hnh nghin cu
Nghin cu v dc liu bin l mt lnh vc cn mi m i vi cc nh khoa
hc trong nc. ti chn cch tip cn, phng php nghin cu v cc k thut
hin i thc hin ti. C th nh sau:

 Cch tip cn nghin cu ca ti:

M hnh nghin cu la chn trong ti l m hnh nghin cu ho hc theo
nh hng hot tnh sinh hc (bioassay-guided chemical study). y l m hnh nghin
cu khp kn, t thc tin nghin cu n c bn trin khai, to sn phm c th phc v
cho cuc sng bng con ng ngn nht, nng cao gi tr s dng v gp phn bo v
ngun ti nguyn thin nhin. S tm tt cc bc tin hnh ca m hnh ny c
trnh by s 1:
Mu sinh vt bin x l
Chit bng MeOH 90%
Dch MeOH th

Khng c hot tnh

Dng li hoc
th hot tnh
khc

Th hot
tnh
C hot tnh
Dch chit MeOH
- Tch phn on
(Chit dung mi phn cc khc nhau hoc sc k ct)

Phn on 1

Phn on 2

Phn on 3

Th hot
tnh

Th hot
tnh

Th hot
tnh

Khng c hot tnh

Dng li

Khng c hot tnh

C hot tnh
Tch cht sch v tinh ch
ch bng TLC/CC/HPLC

Cht 1

Cht 2

Th hot
tnh

Th hot
tnh

Khng c hot
tnh
Dng li

Dng li

Cht 3

Th hot
tnh

Khng c
hot tnh

C hot tnh
Cc nghin cu su
hn v dc l

Dng li

S 1: Nghin cu ho hc theo nh hng hot tnh sinh hc

5

II.1.2. Thit k nghin cu

y l mt ti nghin cu lin ngnh, gii quyt c cc nhim v t ra,
chng ti bn bc v xy dng s nghin cu nh sau:

C s d
liu

X l, lu
gi mu, to
chit phm
phc v
sng lc
HTSH

Thu thp
mu sinh
vt bin.
Xc nh
tn KH v
to tiu bn

Xy dng
cc qui trnh
NCHH theo
nh hng
HTSH

Sng lc
hot tnh
Sinh hc
(Bioassay
sceening)

Cc QT
tch chit,
phn lp
cc cht c
HTSH

Cc kt qu nghin cu
trin khai trong ti KC09.15
Dy chuyn
thit b quy m
Pilot

Thc phm chc nng

HASAMIN tng cng
sinh lc t hi sm

Qui trnh sn xut nguyn liu

Thc phm chc nng

HALIOTIS t bo ng

Thc phm chc nng

CEFISH h tr iu tr
bnh tim mch t
Omega 3

Thc phm chc nng

BIONAMINE t
sao bin

Sn phm thc
thc t phc v
cuc sng

Cc th nghim dc l, c tnh bn trng din,

hiu lc ch phm (HASAMIN, BIONAMINE,
HALIOTIS v CEFISH)

Th nghim v hon thnh quy trnh sn xut

HASAMIN, BIONAMINE, HALIOTIS v CEFISH

Cc th nghim lm sng trn bnh nhn

ng k
sn phm

 T chc thc hin

ti KC09.09/06-10 l ti nghin cu lin ngnh (Ho, Sinh, Y, Dc v
Cng ngh), cc mng nghin cu gn kt vi nhau v kt qu thc hin ca mi nhm
nghin cu u ng vai tr quan trng n kt qu chung ca ti. V vy, ngay t
khi xy dng cng cng nh khi bt u thc hin ti, chng ti tp trung
c lc lng cn b khoa hc v trin khai xy dng tin nghin cu v ni
dung nghin cu thng nht.
1) Vin Ti nguyn v Mi trng Bin Hi Phng
Chu trch nhim thu thp mu sinh vt bin, xc nh tn, phn loi, to tiu bn
mu v cc thng tin v mu. S lng mu, a im thu mu, s chuyn kho st v
cc k thut x l s b, bo qun mu theo yu cu ca ti.
2) Vin Ho hc cc Hp cht thin nhin:
C quan ch tr ti KC09.09/06-10, chu trch nhim chung. Cc phng
nghin cu trong Vin Ho hc cc Hp cht thin nhin tham gia vo cc phn vic
chnh bao gm:
X l mu sinh vt bin, to dch chit phc v cho sng lc hot tnh sinh hc
v nghin cu ho hc. Xy dng qui trnh x l mu thng nht.
Sng lc hot tnh sinh hc ca cc mu sinh vt bin theo cc hng sau:
Hot tnh khng vi sinh vt kim nh (Antimicrobial activity assay)
Hot tnh gy c t bo (Cytotoxic activity assay)
Hot tnh chng xi ho (Antioxidant activity assay)
Nghin cu ha hc theo nh hng hot tnh sinh hc: Tch chit v xc nh
cu trc cc hp cht c hot tnh sinh hc cao t cc i tng chn lc v hng s
dng.
- Xy dng qui trnh tch chit tng hp cc ch phm c th phc v y t
Ch phm HASAMIN t Hi sm.
Ch phm BIONAMINE t Sao bin.
Ch phm HALIOTIS t Bo ng.
Ch phm Thc n chc nng CEFISH phng nga v h tr iu tr bnh tim
mch.
- Tham gia xy dng tiu chun c s cc ch phm trn, thm nh tiu chun
c s ti Cc An ton V sinh Thc phm B Y t
- Xy dng c s d liu v dc liu bin
3) Vin Y hc C truyn Qun i:
- Th c tnh cp (LD50)
-Th c tnh bn trng din
- Th hiu lc ch phm trn ng vt thc nghim
- Hon chnh h s th nghim dc l.
- To ch phm HALIOTIS t Bo ng
4) Vin Cng ngh Sinh hc:
- Thm d kh nng lm dc liu ca vi sinh vt bin, xy dng qui trnh tch
cht c hot tnh
- Phi hp to sn phm BIONAMINE v thng mi ho sn phm
5) Vin Cng nghip thc phm

Xy dng v tin hnh qui trnh sn xut thc phm chc nng CEFISH phng
nga v h tr iu tr tim mch
6) Cng ty Dc phm ng Dng
Sn xut th, ng k lu hnh v thng mi ho sn phm HASAMIN
7) Mt s n v khc
Cng phi hp vi Vin Ho hc cc Hp cht thin nhin trong vic thc hin
mt s cng vic lin quan n ti.
II.2. Cc phng php nghin cu
II.2.1. Phng php thu mu, x l s liu phn tch mu v xy dng c s d liu
II.2.1.1. Phng php thu mu
Mc ny trnh by cc phng php thu mu c p dng gm:
- Theo quy phm thu thp mu sinh vt bin do UBKH KTNN ban hnh nm 1981.
- Phng php thu mu sinh vt trong quan trc v phn tch mi trng bin do B KHCN v
MT ban hnh nm 2002.
- Phng php thu mu sinh vt ca Winkinson v Baker nm 1998.
- S dng phng php ln su kt hp vi cc thit b quay phim chp nh.

II.2.1.2. Cc loi ho cht

Mc ny trnh by cc loi ha cht c s dng nhum mu, nh hnh v
bo qun lu di mu tiu bn.
II.2.1.3. X l s liu v phn tch mu
- Tt c mu thu c u c chp nh trn my nh k thut s v lu gi trong my
vi tnh, su ngm trong cn 70 lu tr lm mu so snh sau ny
- Mu dc gi n cc chuyn gia trong nc v ngoi nc gim nh theo cc
phng php chuyn dng
- S dng phn mm visual basic v Microsoft Acess xy dng c s d liu v cc
loi sinh vt cha dc liu.
- Xy dng c s d liu v cc loi sinh vt cha dc liu
II.2.2 Phng php x l mu, to dch chit v sng lc hot tnh sinh hc
Do c nhng c trng ring, cc mu sinh vt bin i hi phi c bo qun
v x l rt cn thn trnh h hng v mt hot tnh trong qu trnh x l. Phn ny
gii thiu tm tt cch thc xy dng quy trnh x l, to dch chit v bo qun mu
phc v cho sng lc hot tnh sinh hc v xy dng cc protocol thng nht trong nh
gi hot tnh sinh hc. Cc qui trnh v protocol ny c tun th trong sut qu trnh
thc hin d n. Ni dung chnh gm:
II.2.2.1. X l mu v to chit phm phc v sng lc hot tnh sinh hc
Chi tit cc cng on phi c tin hnh x l mu, to dch chit v bo
qun mu c trnh by trong mc ny. Tun th nghim ngt cc quy trnh ny s
m bo c chnh xc cao, trnh nhm ln gia cc mu hoc chit xut khng
trit cc hot cht gip nh hng mt cch chnh xc cho cc nghin cu tip theo.

II.2.2.2 Cc phng php sng lc hot tnh sinh hc

a) Quy trnh nh gi hot tnh khng vi sinh vt kim nh (Antimicrobial activity
assay):
Hot tnh khng vi sinh vt kim nh c nh gi trn cc phin vi lng 96
ging (96-well microtiter plate) theo phng php hin i ca Vander Bergher v
Vlietlinck (1991), v McKane, L., & Kandel (1996), hin ang c p dng ti trng
i hc Dc, i hc Tng hp Illinois, Chicago, M. Cc chng vi sinh vt kim nh
c s dng bao gm:
- Vi khun Gr (-): Escherichia coli , Pseudomonas aeruginosa
- Vi khun Gr (+): Bacillus subtillis, Staphylococcus aureus.
- Nm si: Aspergillus niger, Fusarium oxysporum.
- Nm men: Candida albicans, Saccharomyces cerevisiae.
Thc nghim c tin hnh ti Phng Sinh hc thc nghim, Vin Ha hc cc
Hp cht Thin nhin.
b) Quy trnh nh gi hot tnh gy c t bo (Cytotoxic assay):
Hot tnh gy c t bo c nh gi theo phng php ca Skehan v cng s
(1990) v Likhiwitayawuid v cng s (1993) ti Phng sinh hc thc nghim, Vin
Ha hc cc HCTN. Cc dng t bo ung th ngi c s dng l:
- Dng KB (Human epidemoid carcinoma - Ung th biu m) t phng th
nghim Bioassay trng i hc Dc Illinois- USA.
- Dng Fl (Fibril sarcoma of Uteus - Ung th mng t cung).
- Dng Hep-G2 (Hepatocellular carcinoma- Ung th gan) t Vin VSDT T.
c) Quy trnh th hot tnh chng xy ho ca cc mu sinh vt bin thng qua phn ng
bao vy gc t do DPPH (DPPH radical scavenging activity)
Th nghim bao vy cc gc t do to ra bi DPPH (1,1-diphenyl-2picrylhydrazyl) ca cc cht chit da trn nguyn l: DPPH c kh nng to ra cc gc
t do bn trong dung dch EtOH bo ho. Khi cho cc cht th nghim vo hn hp ny,
nu cht c kh nng lm trung ho hoc bao vy cc gc t do s lm gim cng
hp ph nh sng ca cc gc t do DPPH. Hot tnh chng xy ho c nh gi da
trn gi tr hp ph nh sng ca dch th nghim so vi i chng khi c trn my
Elisa bc sng 515nm. Thc nghim c tin hnh ti Phng Sinh hc thc nghim,
Vin Ha hc cc Hp cht Thin nhin.
d. nh gi hot tnh chng long xng thng qua cc c ch kch thch s bit ha
ca nguyn bo xng MC3T3-E1.
Hot tnh chng long xng c nh gi bng nh hng ca mu th i vi
s sinh trng ca t bo, hot lc ca enzim ALP (Alkaline Phosphatase), hm lng
collagen v v s tch t canxi i vi t bo MC3T3-E1, mt tin nguyn bo xng,
c chp nhn rng ri lm m hnh in vitro nghin cu s bit ha ca nguyn bo
xng ti trng i hc Dc, i hc Quc gia Chungnam, Hn Quc.
e. nh gi hot tnh khng NF-B.
Cc th nghim hot tnh khng NF-B c tin hnh theo phng php SEAP
ASSAY trn dng t bo chun RAW264 ti Phng nghin cu ung th, Vin Sinh hc
v Cng ngh sinh hc Hn Quc (KRIBB).
II.2.3. Phng php phn lp v xc nh cu trc ho hc
Cc phng php cu ch yu gm:
- Cc phng php sc k: Sc k lp mng (TLC), sc k ct (CC)...
- Cc phng php ph bao gm: Ph hng ngoi (IR), ph khi lng (ESI-MS), ph
khi lng phn gii cao (HR FAB-MS), ph cng hng t ht nhn (NMR)...

II.2.4. Nguyn liu v phng php nghin cu thm d kh nng khng vim t
rong to bin Vit Nam
II.2.4.1. Nguyn liu
a. Mu rong
Danh sch 22 mu rong to bin c s dng trong nghin cu do cn b Phng
Cng ngh To, Vin CNSH v CN. Hunh Quang Nng, Vin nghin cu v ng dng
Cng ngh Nha Trang (VAST) thu thp v nh tn khoa hc. Thi gian thu mu 20072008. Trong , Undaria pinnatifida, Enteromorpha linza l hai loi c hot tnh khng
vim cao do Trng i hc Quc gia Pukyong, Hn Quc cung cp c s dng lm
mu th so snh vi rong bin Vit Nam.
Danh sch 22 mu rong bin c dng tch dch chit c hot tnh khng vim
TT
1

Tn mu
Gracilaria asiatica

G. blodgettii

3
4
5
6
7

Sargassum duplicatum
Ulva reticulata (1)
Sargassum polycystum
Codium arabicum
Enteromorpha
compressa
Ulva reticulata (2)
Dictyota dichotoma
Sargassum microcustum
S. mcclurei
S. denticarpum
S. swartzii
S. binderi
Gracilaria arcuata
G. eucheumoides
G. salicornia
Hypnea valentina
Porphyra crispata
P. suborbiculata
Undaria pinnatifida
Enteromopha linza

8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

a im thu mu
m nui trng thu sn, Trm Nghin cu nui trng
thu sn nc l Qu Kim, Hi Phng
m nui trng thu sn, Trm Nghin cu nui trng
thu sn nc l Qu Kim, Hi Phng
Ca l, Ngh An
Do Cng ty TNHH Vit i cung cp
Do Cng ty TNHH Vit i cung cp
Nha Trang, Khnh Ho
Nha Trang, Khnh Ho
Nha Trang, Khnh Ho
Nha Trang, Khnh Ho
o tri Nguyn, Nha Trang, Khnh Ho
Hn Chng, Vnh Phc, Nha Trang, Khnh Ho
M Ho, Ninh Hi, Ninh Thun
Bi Tin, Vnh Ho, Nha Trang, Khnh Ho
Tri Bu, Vnh Nguyn, Nha Trang, Khnh Ho
Bi Tin, Vnh Ho, Nha Trang, Khnh Ho
Nha Trang Khnh Ho
Nha Trang Khnh Ho
Nha Trang Khnh Ho
Tha Thin Hu
Tha Thin Hu
i hc Quc gia Pukyong, Hn Quc
i hc Quc gia Pukyong, Hn Quc

b. ng vt th nghim
+ Chut cng trng (c v ci) kho mnh c trng lng khong 120 20g do
Trung tm chn nui ca Hc Vin Qun Y cung cp.
+ Chut nht c c trng lng khong 22-25 gam do Vin V sinh dch t
Trung ng cung cp.
Cc ng vt th nghim c nui trong iu kin ng nht ca phng th
nghim c y thc n v nc ung trc khi dng lm th nghim.
II.2.4.2. Phng php nghin cu
a. Quy trnh tch chit dch c hot tnh khng vim
Dch chit c hot tnh khng vim trong cc mu to nghin cu c tch chit
theo phng php ca Jin v cs, 1997 v Khan 2007.
10

b. Phng php th hot tnh khng vim trn tai chut

Theo phng php ca Van Arman (1974).
c. Phng php th hot tnh khng vi sinh vt kim nh (pha long nng )
- Pha long mu th bng phng php pha long a nng : Mu ban u c
nng 20mg/ml c pha long thnh cc nng khc nhau th hot tnh vi cc
chng c nng 128 g/ml; 32 g/ml; 8 g/ml; 2 g/ml; 0,5 g/ml i vi cht sch.,
256 g/ml; 64 g/ml; 16 g/ml .. 1 g/ml; 0,25 g/ml i vi dch chit.
d. Nghin cu c tnh cp
Nghin cu c tnh cp v xc nh LD50 ca dich chit c hot tnh khng vim
t Sargassum swartzii (S) v Ulva reticulata (U) trn chut nht trng bng phng
php ca Litchfield v Wilcoxon (1949)
e. Nghin cu tc dng gim au
Tc dng gim au c xc nh bng phng php Koster - qun au axit acetic
v phng php mm nng:
* Phng php gy vim cp
+ Gy ph chn chut bng carrageenin.
+ Gy vim mng bng
* Gy vim mn (gy u ht)
f. X l s liu
Cc s liu nghin cu c x l thng k theo phng php t-test Student v test
trc - sau(Avant - Aprs). S khc bit c ngha khi p < 0,05.
II.2.5. Nguyn liu v phng php nghin cu thm d kh nng sinh cc cht c
Hot tnh sinh hc t vi sinh vt bin
II.2.5.1. Nguyn liu
- Cc mu nc v trm tch c ly cc a im khc nhau ca bin Vit Nam:
Qung Bnh, Bnh nh, Ph Yn vo thng 5/2007. Cc mu ny c Vin a L
cung cp.
- Cc mu san h v rong ly t bin Nha Trang, o Trng Sa 6/2007 do Phng Sinh
thi nc Trung tm Nhit i Vit Nga cung cp. Cc mu nc bin, trm tch bin
su > 2m tr ln c ly bng cc dng c chuyn dng.
- Cc mu nc b mt bin Hi Phng, Ct B, Nha Trang, H Long, Nam nh do
nhm ti thu thp thng 6-7/2007.
- Cc mu c bin, ngao, s huyt mua ti ch Thnh Cng, H Ni vo thng 5/2007.
- Su t (Plutella xylostella), su xanh (Heliothis armigera) do Vin Bo v Thc vt
cung cp.
- Giun nhiu t Derodigitata miiler (thc n cho c cnh) mua ti ch H Ni.
- 5 chng VSV kim nh nhn t bo tng ging chun ca Trung tm CNSH, i hc
khoa hc t nhin H Ni: vi khun Gram (-): E. coli; Pseudomonas aeruginose, vi
khun Gram (+): Bacillus subtilis; Streptococcus aureus, Nm men gy bnh: Candida
albicans.
II.2.5.2. Mi trng phn lp vi sinh vt (g/l)
- Mi trng MPA (phn lp vi khun) Cao tht: 5; pepton: 10; NaCl: 5; Thch: 20;
Nc bin 100%.
- Mi trng Gause 1 (phn lp x khun) Glucoza :10; K2HPO4: 0,5; MgSO4: 0,5;
KNO3: 1; FeSO4: 0,01; Thch: 20; Nc bin.
- Mi trng Hansen (phn lp nm men) Glucoza: 50; pepton 5; MgSO4.7H2O: 3;
K2HPO4: 3; KH2PO4: 3; Cao nm men: 1; Thch: 20; Nc bin.
11

- Mi trng Czapek (phn lp nm mc) Glucose: 5: MgSO4: 0,5; KCl: 0,5; MgCl2: 3;
FeSO4: 0,01; NaNO3: 3; KH2PO4: 1; K2HPO4 : 1; Thch: 20; Nc bin. Tt c cc mi
trng trn u c b sung dung dch nguyn t vi lng.
II.2.5.3. Phng php nghin cu
- S lng vi sinh vt trong cc mu phn tch c xc nh theo phng php
pha long ti hn ca Koch.
- Sng lc s b kh nng sinh cht khng khun, khng nm ca cc chng
phn lp c trn a thch MPA cha 2% dch VSV kim nh.
- Nhum Gram: lm tiu bn cc chng vi khun c hot tnh sinh hc.
- M t hnh thi khun lc bng phng php cy khun lc ln trn a thch.
- Hot ho cc chng vi khun kim nh trn mi trng MPA 370C trong
vng 24 gi, cn chng nm men C. albicans trn mi trng Hansen 300C trong 48
gi.
- Nui cy VSV bin th kh nng sinh cc cht c HTSH.
- Xc nh kh nng khng cc VSV kim nh bng phng php c l v o
vng v khun.
- Phng php xc nh kh nng gy c t bo trn giun t Derodigitata
miiler.
- Phng php th kh nng dit su hi.
- Xc nh hot tnh amilaza trn mi trng cha 1% tinh bt tan; hot tnh
xenluloza trn mi trng cha 1% bt giy v hot tnh proteaza trn mi trng cha
1% cazein. Dung dch hin mu l Lugol 1%.
- Xc nh nh hng ca ngun cacbon, nit, pH v thong kh ln sinh
trung v sinh tng hp cc cht khng khun, khng nm theo phng php VSV thng
dng.
- nh tn khoa hc cc chng vi sinh vt bin c hot tnh sinh hc theo phng
php hnh thi, sinh l sinh ho v sinh hc phn t.
II.2.6. Nguyn liu v phng php nghin cu to thc phm chc nng b sung
omega-3 phng v h tr iu tr bnh tim mch
II.2.6.1. Phng php nghin cu
a. Phng php cng ngh
- Phng php p n trn my p n VTP 2006 do Vin Cng nghip Thc phm
thit k.
b. Phng php phn tch ho l
- Xc nh hm m theo phng php sy n 1050C trong 3 gi n trng lng
khng i.
- Xc nh m ton phn bng phng php Kieldahn.
- Xc nh hm lng bo bng phng php Sochlex.
- Xc nh n: n ca bng c xc nh bng t s gia din tch ca
bng/din tch u khun.
- Xc nh mu (L,a, b) bng my Hunter M.
- Xc nh nht trn my o nht Brookfield DV-II.
- Hm lng DHA c xc nh ti Vin Dinh dng.
- Xc nh axt bo omega 3 trn my sc k kh: cht bo c methyl ho, chy trn
my GC PLUS HP 6890 ca Vin Nghin cu Dc phm v M phm min Nam.
c. Phng php vi sinh
- Xc nh tng s vi khun hiu kh trn mi trng thch tht, pepton.
- Pht hin E. coli trn canh thang thng, canh thang lactoza, nc pepton.
12

- Pht hin vi khun Salmonella trn mi trng EMB (eosin methylen blue).
- Pht hin nm mc, nm men trn mi trng thch malt.
d. Phng php th nghim lm sng
i tng th nghim l cc bnh nhn c cc yu t v nguy c ca bnh tim mch
nh cao huyt p, ri lon chuyn ho lipit mu ang c iu tr ti Bnh vin a
khoa tnh H Ty.
e. Phng php nh gi cm quan: theo TCVN
f. Phng php ton hc
Qui hoch ton hc thc nghim ti u ha cc iu kin cng ngh ca qu trnh
p n bng ma trn ton hc thc nghim Doehlert.
II.2.6.2. Thit b, dng c, nguyn vt liu v ho cht s dng cho nghin cu
Mc ny trnh by cc thit b, nguyn vt liu v ha cht c s dng trong
nghin cu sn xut thc phm chc nng b sung Omega-3 phng v iu tr bnh tim
mch.
II.2.7. Phng php nghin cu to thc phm chc nng Bionamine tng cng
sc khe cho vn ng vin t sao bin
II.2.7.1. Nghin cu xc nh hm lng hot cht sinh hc hoocmn steroit
(testosteron, progesteron, cortilson, oestradoen) trong cc i tng nghin cu (theo
phng php min dch Eliza trn my Multiskan - Phn Lan vi kit ca hng Biotech
M).
II.2.7.2. Nghin cu xc nh hm lng hot cht sinh hc, cc nguyn t khong vi
lng cn thit (Fe, Zn, Mn, Se) trong cc i tng nghin cu (bng phng
php phn tch hunh quang tia X trn my k hiu XRF SEA-2110 ca Nht).
II.2.7.3. Nghin cu xc nh hm lng hot cht sinh hc (protein, axit amin) ca
cc i tng nghin cu (bng phng php in di trn my axt amin t ng HP
Amino Quant Series II ca Nht).
II.2.7.4. Nghin cu quy trnh khai thc cc hot cht sinh hc t to ch phm thc
phm chc nng vin tng lc Bionamine bng cng ngh enzim.
* Phn tch hot cht sinh hc trong mt s loi sao bin bng phng php sinh
ha, phng php min dch Eliza, phng php vi sinh v phng php phn tch hunh
quang tia X.
* ng dng cng ngh enzyme, cng ngh ha sinh to cc sn phm bt nguyn
liu Bionamine.
* Phng php xc nh mc thu phn c cht: Phng php Auson ci tin, v
phng php chun formol.
II.2.7.5. Nghin cu c tnh cp v c tnh bn trng din ca bt Bionamine.
Cc nghin cu thc nghim v c tnh cp v bn trng din c thc hin ti
Trung tm nghin cu c hc Hc Vin Qun Y.
II.2.7.6. Phng php th nghim tc dng ca vin tng lc Bionamine trn ng
vt thc nghim.
Tc dng ca vin tng lc Bionamine trn ng vt thc nghim c nh gi
thng qua cc ch tiu huyt hc, tng cng th lc v sc bn ca chut trong qu
trnh bi ln.
II.2.7.7. Phng php nh gi tc dng lm sng ca vin Bionamine trn vn ng
vin in kinh
nh gi tc dng v sc bn, sc nhanh (phn x thn kinh) v sc mnh c bp
trn vn ng vin ca cc i in kinh H Ni.

13

II.2.8. Nguyn liu v phng php nghin cu sn xut vin nang mm bo ng

HALIOTIS
II.2.8.1. Phng php nghin cu qui trnh bo ch vin nang Haliotis
a. Ni dung nghin cu
- Nghin cu la chn dc liu v xy dng qui trnh bo ch vin nang mm Bo ng
HALIOTIS thnh dng bn thnh phm v thnh phm
- Nghin cu xy dng tiu chun c s cho sn phm vin nang mm bo ng
HALIOTIS
- Kim nghim sn phm vin nang mm HALIOTIS theo tiu chn c s xy dng
- Tin hnh ng k sn phm di dng thc phm chc nng vi B y t xin cp giy
php lu hnh ton Quc.
b. Nguyn liu v phng php nghin cu:
+ Nguyn liu:
Tht bo ng c ly t vng bin Nha Trang do Trung tm nhit i Vit Nga
chi nhnh Nha Trang cung cp vi s lng 30 kg.
+ Cch x l:
- Bo ng sau khi thu hoch tch b v cng loi ht tp cht v ph tng, ra sch
x l bo qun ng kh.
- Thi lt mng, sy cho n kh
- Nghin thnh bt mn
+ Phng php nghin cu:
* Nghin cu la chn dc liu v xy dng qui trnh bo ch vin nang mm Bo ng
Haliotis:
- Bo ng c la chn t theo tiu chun qui nh ca Dc in c th: T l
thu phn khong 73%, 24,58% protit, 0,44% cht bo, 1,98% tro.
- Xy dng qui trnh bo ch vin nang mm Haliotis:
- Bo ng sau khi thu hoch tch b v cng loi ht tp cht v ph tng, ra sch
x l bo qun ng kh.
- Thi lt mng, sy cho n kh
- Nghin thnh bt mn
- Nh ho bng du u lnh thu c dng bn thnh phm
- ng nang mm Gelatine trn dy chuyn ng nang mm t tiu chun GMP
WHO
- Sy vin
- ng l hoc p v, dn nhn hon thin sn phm
- Kim nghim thnh phm
- Nghin cu
II.2.8.2. Phng php nghin cu c tnh cp
Nghin cu c tnh cp c thc hin ti Khoa Nghin cu thc nghim - Vin
Y hc c truyn Qun i theo cc phng php:
- Phng php xc nh c tnh cp ca thuc Nh xut bn Y hc, 1996.
- Hng dn km theo quyt nh s 371/BYT Q ngy 12/3/1996 v xc nh an
ton cho thuc c truyn ca B Y t
- General guidelines for methodologies on research and evaluational of traditional
medicine. World Health Organization 2000
- Principles and Methods of Toxicology A. Wallace Hayes, 1994.
II.2.8.3. Phng php nghin cu c tnh bn trng din
Mu th: Liu th: 2 vin/ kg/ ngy
14

Ct vin nang mm, p ra ht cht dch v thm nc, trn u thu c hn

dch c cha lng thuc tng ng 2 vin/ 5ml. Khuy trn u hn dch mi khi ly
cho th ung.
Chun b mu th hng ngy trc khi cho th ung.
B tr th nghim v tin hnh: Th nghim c tin hnh trn 20 th, chia lm hai
nhm chng v th, mi nhm 10 th.
- Nhm chng: ung nc ct 5,0 ml/ kg/ ngy
- Nhm th : ung 5 ml dch mu th chun b nh trn vi liu 0,3 g/ kg/ ngy
Cho th ung hng ngy, lin tc trong mt thng.
Th c 2 nhm c nui dng trong cng iu kin v cng ch dinh dng.
Theo di v nh gi: Theo di th hng ngy v mc tiu th thc n, kh nng
hot ng, tnh trng phn, lng.
Trc khi th nghim, xc nh ch s bnh thng ca th v cn nng, cc du
hiu ton thn, cc ch s v huyt hc, ho sinh chc nng gan, thn. Ht thi gian th
nghim, nh gi li th trn cc ch tiu nh trc th nghim. M th quan st i th
cc t chc gan, thn, phi, tim v hnh dng, mu sc, th cht, tnh trng xung huyt...
So snh kt qu trc v sau th nghim ca nhm chng, nhm th, gia nhm
chng v nhm th theo phng php thng k y sinh hc.
II.2.8.4. Phng php nghin cu tc dng bi b c th trn lm sng
a. Cht liu nghin cu:
Vin nang mm Haliotis bo ch t Bo ng c ng gi 170 mg/ vin, do
khoa dc - Vin YHCT Qun i sn xut.
b. i tng nghin cu
Gm 45 bnh nhn (BN) t nguyn 40 tui
+ Tiu chun chn bnh nhn
- Bnh nhn 40 tui, khng phn bit gii, ngh nghip.
- Bnh nhn c chn on l suy nhc c th theo tiu chun chn on ca CDC
nm 1994.
- Theo YHCT bnh nhn thuc chng h lao th can thn m h: au u, hoa mt,
chng mt, tai , mt ng, hay qun, th trng gy; ngi mt mi, hay cu gt, lng
bn tay chn nng; ra m hi trm; au lng; mi gi; nhc trong xng.
- Bnh nhn chp nhn dng thuc, tham gia cng trnh nghin cu, thc hin nhng
iu quy nh trong qu trnh nghin cu.
- Bnh nhn iu tr ni tr ti khoa lo khoa v iu tr ngoi tr ti phng khm bnh
vin YHCT Qun i.
+ Tiu chun loi tr
- BN ang dng thuc khc hoc suy thn, suy gan, suy tim, thiu mu nng, bnh thn
kinh thc tn
- BN khng hp tc nghin cu.
c. Phng php nghin cu
- Phng php nghin cu: Theo phng php th nghim lm sng so snh trc v
sau iu tr.
- Thit k nghin cu: 45 bnh nhn p ng tiu chun nghin cu. Thi gian nghin
cu 4 tun.
- Phng php dng thuc: dng vin nang mm bo ng 2 vin/ln x 2 - 3 ln/ngy x
30 ngy iu tr ni tr ti Khoa Lo khoa v iu tr ngoi tr ti phng khm bnh Vin YHCT Qun i.

15

- Tt c cc bnh nhn nghin cu u c khm xt t m v ni khoa, lm bnh n

c chn on theo YHCT v y hc hin i, c phiu theo di, thm khm hng ngy
theo di kt qu v ghi tc dng ph khng mong mun ca thuc.
II.2.9. Phng php nghin cu sn xut thc phm chc nng vin nang mm hi
sm HASAMIN
II.2.9.1. Phng php xc nh cc thnh phn dinh dng, hoocmon v khong
cht trong Vin nang hasamin
a. Nghin cu xc nh hm lng hot cht sinh hc hoocmn steroit (testosteron,
progesteron, cortilson, oestradoen) trong cc i tng nghin cu theo phng php
min dch Eliza trn my Multiskan - Phn Lan vi kit ca hng BiotechM.
b. Nghin cu xc nh hm lng cc nguyn t khong vi lng cn thit (Fe, Zn,
Mn, Se) trong vin Hasamin bng phng php phn tch hunh quang tia X trn my
k hiu XRF SEA-2110 ca Nht.
c. Nghin cu xc nh hm lng hot cht sinh hc (protein, axit amin) ca trong
vin Hasamin bng my axt amin t ng HP Amino Quant Series II ca Nht.
II.2.9.2. Phng php th nghim c tnh cp v c tnh bn trng din
c tnh cp v c tnh bn trng din ca vin nang mm hi sm Hasamin
c nghin cu ti Vin Y hc C truyn Qun i theo cc phng php c trnh
by mc II.2.7.2 v II.2.7.3.
II.2.9.3. Phng php th nghim tc dng ca vin Hasamin trn ng vt
i tng:
Th nghim tin hnh i vi 10 th c thuc ging Newzealand, nui ti Trung
tm Nghin cu D v Th Sn Ty. Th ging c trng lng 3,0 - 5,0 kg, tui 12 - 26
thng, kho mnh, giao phi thun thc, nui nhng chung ring bit, ch nui dng bao gm thc n tinh hn hp 16% protein 80g/ngy v 600g thc n xanh.
Phng php:
Ly tinh th bng m o gi, nhit m o gi 45oC, ly tinh vo bui sng.
Th tch tinh dch (V), hot lc tin thng ca tinh trng (A), nng tinh trng (C), sc
khng (R) i vi dung dch NaCl 1% ca tinh trng xc nh theo phng php ca
Milovanov (1962), tng s tinh trng tin thng ca mt ln ly tinh (V.A.C) xc nh
theo phng php ca Herrick v Self (1962), t l tinh trng sng (LS) theo phng
php nhum mu phn bit vi eosin 5% ca Morozov (1938), t l tinh trng k hnh
(K) theo phng php nhum mu vi fuxine ca Serdiuc (1970), pH tinh dch o bng
giy o pH ca Trung Quc.
M hnh th nghim:
L 1: 05 con (s tai:4182; 813; 853; 833; 91531), th n theo khu phn c s +
HASAMIN.
L 2: 05 con (s tai:94514; 11558; 1471; 1000; 836), th n theo khu phn c s
+ vin Placebo (Vin trng)
Thi im nh gi phm cht tinh dch:
Trc khi cho n vin HASAMIN, Placebo.
Sau 10 ngy b sung vin HASAMIN, Placebo.
Sau 20 ngy b sung vin HASAMIN, Placebo.
Liu lng v cch cho n: Liu lung b sung: 01 vin/con/ngy. Cch b sung
vin thuc: trn u thuc vi mt t thc n tinh, cho th n ht, sau mi cho n thc n
c s.
16

Phn II. kt qu nghin cu

A. Kt qu Thu thp mu sinh vt bin v sng lc
hot tnh sinh hc
Chng I. Thu thp, nh loi mu sinh vt bin v xy dng
c s d liu
I.1. tnh hnh nghin cu nhm Hi min, Da gai, San h mm
Phn ny trnh by mt cch vn tt cc nghin cu thu thp, nh loi, v phn b
ca cc loi sinh vt bin thuc ba nhm hi min, da gai v san h mm.
I.2. a im v thi gian thu mu
ti t chc 9 chuyn thu mu (Ct b thng 11/2006; Cn C thng
11/2006; Trng Sa thng 3-4/2007; Cn o thng 6/2007; Vnh H Long thng
7/2007 ; Ct B thng 8/2007; Cn C t 6 n 10/6/2008; Ct B t 27/ 6 n
5/7/2008; Vnh H Long t 9/9 n 16/9/2008). Tng s mu thu c trong ti ny
l 101 mu sinh vt Bin. C th bn cc t thu mu c trnh by trn hnh di
y:

Vnh H
Long
Ct B

Cn
C

Trng
Sa
Cn o

Cc a im thu mu nm 2006 - 2008.

17

I.3. Mt s nt c bn v c im mi trng cc a
im nghin cu
Mc ny trnh by mt s nt c bn v c im mi trng ti cc a im ly
mu: Ct B, Cn o v Trng Sa.
I.4.Kt qu thu thp mu
Trong ba nm 2006-2008, qua 9 t thu mu ti tin hnh thu thp c 101
mu sinh vt Bin c kh nng cha dc liu ti cc a im nh sau:
I.4.1. Thu thp mu t 1 ti Ct B thng 11/2006
- Thi gian thu mu: t ngy 5 n 15/11 nm 2006
- Cc im thu: Ct Da, Vn Bi, Ba Tri o, ...
- S mu thu: 6 mu.
I.4.2. Thu mu t 2, Cn C thng 11/2006
- Thi gian thu mu: t ngy 5 - 20/11/2006
- Cc im thu: o Cn C - Qung Tr
- S mu thu: 9 mu
I.4.3. Thu mu t 3 Trng Sa thng 3-4/2007
- Thi gian thu mu: t 20 thng 3 n 20 thng 4/2007
- Cc im thu: Trng Sa Ln, Nam Yt, Tc Tan
- S mu thu: 6 mu
I.4.4. Thu mu t 4 ti Cn o thng 6/2007
- Thi gian thu mu: t 20 thng 5 n 10 thng 6/2007
- Cc im thu: Hn Tre, Hn By Cnh, Hn Trc, Vnh Cn Sn.
- S mu thu: 17 mu
I.4.5. Thu mu t 5 ti Vnh H Long thng 7/2007
- Thi gian thu mu: T ngy 3 - 15 thng 7/2007
- Cc im thu: u B, Hang Trai, Hang D (Ang D), Cng
- S mu thu: 16 mu
I.4.6. Thu mu t 6 ti Ct B thng 8/2007
- Thi gian thu mu: T ngy 1 - 10 thng 8/2007
- Cc im thu: Ct Da, Vn Bi, Ba Tri o, ...
- S mu thu: 17 mu
I.4.7. Thu mu t 7 ti Cn C t 6 n 10/6/2008
- Thi gian thu mu: T ngy 6 - 10 thng 6/2008
- Cc im thu: Bc Cn C
- S mu thu: 9 mu
I.4.8. Thu mu t 8 ti Ct B t 27/ 6 n 5/7/2008
- Thi gian thu mu: T ngy 27/6 5/7/2008
- Cc im thu: Vn Bi, Ba Tri o, Hang Trai, u B,..
- S mu thu: 7 mu
I.4.9. Thu mu t 9 ti Vnh H Long t 9/9 n 16/9/2008
- Thi gian thu mu: T ngy 9/9 16/9/2008
- Cc im thu: Vn H, ng D, Cng , B Xm, Cc 8, Cc 6
- S mu thu: 14 mu
Trong s 101 mu thu c c 41 mu hi min, chim 40,6 % tng s mu thu
c, 31 mu san h mm (30,7 %) v 29 mu da gai (28,7 %). Trong s :
Hi Min
+ S lng: 41 mu
18

+ V d v s phn b cc mu hi min ti cc im thu mu:

- Cc kt qu phn tch ca chng ti xc nh c 6 mu ti khu vc ng
nam o Ct B (Gellius varius, Dysidea cinerea, Haliclona sp, Mycale plumosa,
Ircinia echinata, Amorphinopsis excavans)
- 4 mu cng tm thy o Trng Sa (Petrosia nigricans, Janthella sp,
Xestospongia testudinaria, Ircinia sp)
- 7 mu Hi min ti vng bin Cn o (Clathria vulpina, Petrosia nigricans,
Xestospongia testudinaria, Niphates sp, Spongia sp, Stylissa flabelliformis, Gellius var
fibrosa)
- Ti Vnh H Long cng thu c 7 mu (Haliclona subarmigera, Dysidea
cinerea, Ircinia echinata, Biemna cf. nudelringna, Xestospongea testudinaria, Gelliodes
fibulata, Aka mucosa)
San h mm
+ S lng: 31 mu
+ V d v s phn b cc mu san h mm ti cc im thu mu:
- Ti Cn C tm thy 7 mu: Cladiella sp, Sarcophyton trocheliophorum,
Lobophytum crassum, Lobophytum sp, Lobophytum compactum, Sinularia abrupta,
Klyxum sp
- Ti o Nam Yt thuc Trng Sa thu c 1 mu san h Lobophytum
compactum
- Ti Cn o thu c 5 mu: Sarcophyton elegans, Sarcophyton mililatensis,
Lobophytum compactum, Sinularia sp, Dendronephthya klunzeringi
- Ti H Long mi ch thu c 3 mu nm trong hai ging Sarcophyton v ging
Cladiella
- Ti Ct b thu c 3 mu thuc hai ging Sarcophyton v ging Cladiella.
Da Gai
+ S lng: 29 mu
+ V d v s phn b cc mu da gai ti cc im thu mu:
- Ti Cn o: thu c 3 mu gm: Hi sm en (Holothuria atra), ui rn
(Macrophiothrix sp), cu gai hnh trng (Echinocardium cordatum)
- Vnh H Long: S lng mu thu c nhiu hn khu vc Cn o, tng s 5
mu gm: Sao bin 5 cnh mu tm ru (Asterope carinifera), sao bin cnh ngn
(Anthenea pentagonala), sao bin 5 cnh mu tm (Asterope carinifera), sao bin 5
cnh mu hng (Protoreaster nodusus), c gim gai di (Diadema setosum)
- Vng bin Ct B : Trong t thu mu thng 8/2007 thu c 6 mu Chng
bao gm hi sm gai (Holothuria spinifera), sao bin rnh nng (Astropecten
polyacanthus), da chut bin mu hng khng gai (Thyone bicornis), da chut bin
mu hng c gai (Stichopus sp), sao bin 5 cnh mu tm ru (Asterope sp) v sao bin
cnh ngn.
Kt qu: thi gian, a im thu thp mu v gim nh tn khoa hc c trnh
by chi tit di dng bng trong bo co chnh.
1.5. M t cc loi sinh vt bin in hnh c kh nng cha
dc liu
Mc ny a ra mt s c im nhn dng ca mt s i tng sinh vt bin
in hnh thuc cc nhm san h mm, da gai v hi min. Do khun kh ca bo co
c hn, chng ti ch trnh by di y nh mu lu v m t ca mt s loi sinh vt
bin in hnh:
19

I.5.1. Cc loi san h mm

1. Loi Lobophytum compactum, thuc h Alcyoniidae

c im: Tp on c dng ph
dy, trn polyparium c cc thu
mc gn nhau, di 3-5cm hu nh
khng phn nhnh. Polyp c b
mt trn ca tp on v trn cc
thu
Mu sc: c mu nu nht, xanh
nu, pht hng

2. Loi Lobophytum crassum, thuc h Alcyoniidae

c im: Tp on c thn ngn
v to hoc dng ph. Polyparium
phn nhnh thnh cc thu hoc
dng mo, rt him khi c dng
phng. Polyp lng hnh. Gai
xng trn b mt c dng di cui,
gai xng trong dng hnh con sut
Mu sc: c mu vng sng, nu
hoc xanh lc.

3. Loi Sarcophyton elegans, thuc h Alcyoniidae

c im: Tp on c mt thn
ln, pha trn dng a mang polyp
gi l polyparium. gia
polyparium c dng lng cho,
pha ngoi gn sng hoc gp khc.
mi trng sng mnh thn ngn
v mp, mi trng yn tnh thn
di v mnh hn
Mu sc: c mu nu sm, vng
nu hoc xanh nu

20

4. Loi Sarcophyton mililatensis, thuc h Alcyoniidae

c im: Tp on c thn ln,
polyparium c dng lng cho, pha
ngoi gn sng hoc gp khc. B mt
tp on nhm, polyp nh. mi trng
sng mnh thn ngn v mp, mi
trng c bo v thn di v mnh hn
Mu sc: c mu vng xanh, nu hoc
nu vng
5. Loi Sarcophyton trocheliophorum, thuc h Alcyoniidae
c im: Tp on c dng nm, thn
ln. B mt polyparium thng gp khc.
Polyp lng hnh. Gai xng trong c hnh
dng con sut vi cc mu li, gai xng
ngoi hnh di cui
Mu sc: c mu vng, nu nht hoc
xanh nht
6. Loi Sinularia abrupta, thuc h Alcyoniidae
c im: Tp on dng ph hoc c
thn phn bit. Polyparium nhn, phn
thu dng ngn hoc dng mo. Polyp n
hnh. Gai xng trong c dng con sut ln
c mu nh, chiu di ln hn 2mm. Gai
xng ngoi dng di cui hoc con sut
nh.
Mu sc: hi trng, vng nht, nu sng
I.5.2. Cc loi hi min
1. Loi Gellius varius, thuc h Chalinidae

M t: dng cnh, mu xanh lc m khi

cn sng, khi cht hoc khi c nh bng
ho cht th chuyn mu xanh en,
chng to thnh khm sng trn cc khi
san h hoc bm trn nn y cng

21

2. Loi Dysidea cinerea, thuc h Dysideidae

M t: mc thnh dng khi trn rn san

h, mu sc thany i rt nhiu ph
thuc vo mi trng sng, t mu nu
nht cho ti mu xm hoc, mm v dai,
thng ln c ct hoc sn bn trong c
th.

3. Loi Haliclona sp, thuc h Chalinidae

M t: thng c dng cnh, cu trc

thay i tu loi c th mm hoc hi
cng, mu sc thay i rt nhiu, t mu
xm nht, hng, tm hoc xanh m, vi
l m trn b mt tng i ln

4. Loi Mycale plumosa, thuc h Mycalidae

M t: c dng cnh c trng v

thng ni vi nhau, mu sc t m
cho ti mu tm nht, thng b mt mu
rt nhanh sau khi ly ra khi mi trng
sng, cu trc tng i mm

22

5. Loi Ircinia echinata, thuc h Irciniidae

M t: thng sng dng khi c x

thu, b mt c bao ph bi cc mu
nh, thng c mu nu en cho ti mu
en, c th do v n hi ging nh cao
su

6. Loi Amorphinopsis excavans, thuc h Halichondriidae

M t: y l loi c l vo trong cc
khi san h hoc nn y v chim c
gn ht khng gian bn trong, mu sc
thay i t vng cho ti vng m, c l
m trn b mt kh ln. Khi kh
thng chuyn mu trng ng

7. Loi Clathria vulpina, thuc h Microcionidae

M t: c th dng cnh, c mu vng

cam, b mt x x c to nn bi nhiu
nm gai nh

23

8. Loi Petrosia nigricans, thuc h Petrosiidae

M t: thng c
dng khi, ging
hnh ni la, cu
trc cng v d v,
d gy, b mt
tng i nhn,
thng c mu tm
nht cho ti nu
m

9. Loi Xestospongia testudinaria, thuc h Petrosiidae

M t: hnh dng,
mu sc v kt cu
b mt thay i rt
nhiu tu thuc
vo mi trng
sng,
thng
chng c dng cc
ln vi b mt
nhiu cc g v
cc rnh, cu trc
cng v d v vn.

10. Loi Niphates sp, thuc h Chalinidae

M t: c th dng
cnh phn nhnh,
mu sc thay i
th mu trng ng
cho ti mu hng
nht, b mt tng
i nhn, c mt
s l m nh trn
b mt.

24

11. Loi Stylissa flabelliformis, thuc h Axinellidae

M t: mc thnh
cm trn rn san
h, c mu vng
cam, b mt x x
thng c nhiu
mu nh hoc c
cc ng g hoc
np gp

12. Loi Ianthella sp, thuc h Ianthellidae

M t: C th c
dng qut, mu sc
thay i t mu
vng nht, mu
vng cam m cho
ti mu tm sm,
khi tip x vi
khng kh thng
chuyn mu tm
sm

13. Loi Ircinia sp, thuc h Irciniidae

M t: thng c
dng x thu, c
mu t nu m ti
mu en, i khi
c bao ph bi
cc mu tht nh,
cu trc do v dai
ging cao su

25

.5.3. Cc loi da gai

1. Hi sm trng Holothuria scabra, thuc h Holothuridae
c im nhn dng: L loi c c th
ln, chiu di c th t n 30 cm. Bnh
thng c th hnh qu chui, nhng c
th thay i rt nhanh nu mi trng c
bin di. C th mu en hoc en hi
chuyn mu nu, mu hng. Trn c th
c hai dng gai hnh gn trn v thoi bao
ph

2. Sao bin cnh dy Protoreaster nodusus, thuc h Oreasteridae

c im nhn dng: Hnh sao 5 cnh,

di cc cnh u nhau v tng i
ngn. Trn mi cnh c t 5 - 6 hnh
trn. Mu sc bin i t mu vng
chanh sang vng nht

I.6. Bc u xc nh cc khu vc c kh nng khai thc

ngun dc liu bin
Cc kt qu thu thp mu vt ca chng ti dc theo chiu di ca bin Vit Nam,
cho php bc u xc nh cc sinh cnh chnh m chng ta c th thu thp
cc loi sinh vt bin c gi tr lm dc liu
1. Cc tng, ng thuc qun o vi nh H Long - Ct B: Thng c nhiu loi
thuc nhm Hi min v sao bin, hi sm, phn b. khu vc ny chng thng
tp trung phn b thnh cc bi ln c gi tr khai thc. y l khu vc c bit cn
lu , trong tng lai, chng c th l nhng a im quan trng cho vic nui trng
v khai thc cc loi c cha dc liu bin. iu ny chng t khu vc H Long Ct B - Bi T Long l nhng ni c cc loi sinh vt cha dc liu tp trung sinh
sng.
2. Rn san h: l sinh cnh quan trng, y c th thu c rt nhiu cc loi sinh vt
c kh nng cha dc liu, c bit l nhm Da gai v san h mm v Hi min.
Cc rn san h thuc ng Nam Ct B, Hang Trai- u B, C T, Hi Vn - Sn
Ch, C Lao Chm, ven b Ninh Thun. c bit 3 khu vc l Cn C, Cn o v
Trng Sa l khu vc c mt cao ca cc loi san h mm.
26

3. Cc bi triu ct hoc ct pha v sinh vt, ct si: Phn b hu ht cc cung lm

ca cc o hoc ven b Vit Nam. y c th thu c hu ht cc loi da gai, hi
min c gi tr dc liu

Hnh I.6. Cc bi dc liu bin ch yu khu vc H Long - Ct B

Ghi ch: 1- Bi Vn Bi - Vn H (Bi sao bin); 2- Bi Tng Gu - Tng Hi (Bi hai mnh v); 3- Bi
Ba Tri o (hi min cnh); 4- Bi Vng Vua - Cng L; 5- ng Nam o Hang Trai; 6- Khu H Ba
Hm

Cc kt qu nghin cu ca chng ti cng xc nhn c 6 khu vc l cc bi

dc liu ln ca Vng bin Vit Nam (v d nh hnh 1.6) l: Bi dc liu ng
nam o Ct B, bi dc liu khu vc vnh H Long, bi dc liu khu vc vnh C T
- Thanh Ln, bi dc liu Cn C, bi dc liu Cn o v bi dc liu Trng Sa.
l cc khu vc c a dng sinh hc cao, t chng ta c th khai thc c cc
loi sinh vt bin c gi tr kinh t v dc liu cao c bit l cc loi thuc nhm hi
min, da gai (sao bin, hi sm, cu gai) v san h mm.
I.7. Xy dng c s d liu sinh vt bin
ti thit k, xy dng c s d liu sinh vt bin cha dc liu. y l
mt c s d liu a dng c xy dng da trn cc phn mm hin i nhm to ra
mt h thng chng trnh h tr cng tc qun l, tra cu, tm kim thng tin v mt s
i tng c cha dc liu sng vng bin ven b nh San h, thn mm, da gai,
rong bin, sa bin. C s d liu c thit k n gin, tin li v d s dng, gip cc
nh khoa hc, nh qun l u c th d dng cp nht, hiu chnh, lit k, b sung
thng tin v cc loi sinh vt cha dc liu vo c s d liu, tm kim v tra cu thng
tin, in n v truy xut thng tin kt qu ra my in.
Cc d liu trong c s d liu c t chc di cc hnh thc sau:
- Cc mu thu c c xc nh c h s mu km theo, c lu tr trong
phng th nghim so snh
- H s ca 101 mu thu (t thu mu, thi gian thu mu, a im v tn loi
thu c)
27

- nh ca tt c 101 mu thu c lu tr bng 101 file dng jpg, c th x l

nhiu phn mm khc nhau
Hin nay, chng trnh c s d liu c trnh by qua h my tnh chuyn
dng ca Vin.
M hnh s dng ca c s d liu c trnh by tm tt trn s sau:

Ngi s dng
Yu cu

Tr li

Giao din ngi s

dng

Kim tra yu cu
ca ngi s
Khi to chc

X l

Hin th thng tin

Cp nht
Tm kim
B sung
Hiu chnh
Hu b
In n thng tin
...

C s d liu
a dng sinh hc

Tng qut chu trnh x l thng tin trong c s d liu

B c s d liu c bin son trn b a CD. Trong c tch hp phn
mm c s d liu sinh vt bin v cc file d liu m t cc sinh vt bin c cha dc
liu. y l mt phn trong nhng sn phm m ti hon thnh giao np cho
Chng trnh KC09/06-10.
Chng II. X l mu, to dch chit v sng lc hot tnh
sinh hc
Tt c cc mu sinh vt bin thu thp c trong ti u c x l s b, bo
qun v to dch chit th theo quy trnh thng nht nh trnh by trong phn phng
php nghin cu. Cc dch chit c tin hnh sng lc hot tnh sinh hc theo nh
hng: khng vi sinh vt kim nh, chng xy ha v gy c t bo. Di y l kt
qu c th:
28

II.1. Kt qu to dch chit

Sau khi chit xong ton b mu, cc s liu ca qu trnh chit mu (K hiu
mu, lng mu chit, lng cn dch thu c) c tp hp li thnh bng v
tnh hiu sut thu hi cn dch. Kt qu chi tit c trnh by trong bng II.1.
Bng II.1. Kt qu to dch chit cc mu sinh vt bin

500
500
500
500
500
1000
1000

Lng
cn
dch
chit
(g)
2,75
2,45
2,80
3,70
4,25
4,00
9,60

1000

13,40

1,34

1000
1000
1000
1000
1000
1000
500
500
500

11,80
15,50
16,50
10,80
8,80
12,20
7,15
3,30
2,85

1,18
1,55
1,65
1,08
0,88
1,22
1,43
0,66
0,57

1000

8,30

0,83

500

4,25

0,85

500

3,75

0,75

Lobophytum compactum
Clathria vulpina

1000

12,10

1,21

500

3,45

0,69

San h mm

Sarcophyton elegans

500

7,75

1,55

KC-024

San h mm

Lobophytum compactum

500

4,85

0,97

KC-025

San h mm hnh l
gan

Sarcophyton mililatensis

1000

10,90

1,09

26

KC-026

San h mm

Lobophytum compactum

1000

13,90

1,39

27
28
29

KC-027
KC-028
KC-029
KC-030

Hi min
Hi min xp
Hi sm en
Hi min hnh qu
u
Da gai ui rn
Hi min xp l
San h mm hnh

Petrosia nigricans
Xestospongia testudinaria
Holothuria atra
Niphates sp

500
1000
500

3,35
8,20
7,60

0,67
0,82
1,52

500

3,95

0,79

Macrophiothrix sp
Spongia sp
Sinularia sp

500
500
1000

3,25
2,05
14,50

0,65
0,41
1,45

K hiu
mu

TnVit Nam

Tn khoa hc

KC-001
KC-002
KC-003
KC-004
KC-005
KC-006
KC-007
KC-008

Hi min cnh xanh

Hi min
Hi min
Hi min
Hi min
Hi min
San h mm
San h mm

KC-009
KC-010
KC-011
KC-012
KC-013
KC-014
KC-015
KC-016
KC-017
KC-018

KC-021
KC-022

San h mm
San h mm
San h mm
San h mm
Hi min
San h mm
San h mm
Hi min
Hi min hnh nn
Hi min hnh
chung
Hi min hnh
chung
Hi min mng
mng
San h mm
Hi min cy

Gellius varius
Dysidea cinerea
Haliclona sp
Mycale plumosa
Ircinia echinata
Amorphinopsis excavans
Cladiella sp
Sarcophyton
trocheliophorum
Lobophyton crassum
Lobophytum sp
Lobophytum compactum
Lobophytum compactum
Amorphinopsis sp
Sinularia abrupta
Klyxum sp
Petrosia nigricans
Ianthella sp
Xestospongia testudinaria

23

KC-023

24

TT

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22

25

30
31
32
33

KC-019
KC-020

KC-031
KC-032
KC-033

Xestospongia testudinaria
Ircinia sp

29

Lng
mu
chit
(g)

% Thu
hi cn
dch
chit
0,55
0,49
0,56
0,74
0,85
0,40
0,96

34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69

KC-034
KC-035
KC-036
KC-037
KC-038
KC-039
KC-040
KC-041
KC-042
KC-043
KC-044
KC-045
KC-046
KC-047
KC-048
KC-049
KC-050
KC-051
KC-052
KC-053
KC-054
KC-055
KC-056
KC-057
KC-058
KC-059
KC-060
KC-061
KC-062
KC-063
KC-064
KC-065
KC-066
KC-067
KC-068
KC-069
KC-070

70
71
72
73

KC-071
KC-72
KC-73

chn vt
Hi min cnh
Hi min phin
San h mm hnh
chm chm
Hi min cnh
Da gai hnh trng
San h mm
San h mm
San h mm
Hi min hnh
chung
C gim gai di
Hi min
Hi min xanh
Hi min hnh ti
Hi min
Sao bin
Hi min
Hi min bt
Hi min trng
Hi min tm
Sao bin nm cnh
mu hng
Sao bin 5 cnh
mu tm
Sao bin 5 cnh
mu tm ru
Sao bin rnh nng
Da chut bin
Hi min xp
Hi min xp
Hi min
Hi sm
Hi min hnh chp
Hi min hnh chp
Hi min
San h mm
San h mm
Sao bin 5 cnh
mu tm ru
Da chut bin mu
hng c gai
Da chut bin mu
hng khng gai
Sao bin cnh ngn
mnh dy mu hng
m
Sao bin
San h mm
San h mm

Clathria vulpina
Stylissa flabelliformis
Dendronephthya
klunzeringi
Gellius var fibrosa
Echinocardium cordatum
Sarcophyton sp
Lobophytum sp
Lobophytum sp
Haliclona subarmigera
Diadema setosum
Ircinia echinata
Biemna cf. nudelringna
Xestospongea testudinaria
Gelliodes fibulata
Anthenea pentagonala
Aka mucosa
Dysidea cinerea
Mycale plumosa
Haliclona submarginera
Protoreaster nodusus
Asterope carinifera
Asterope carinifera
Astropecten polyacanthus
Holothuria spinifera
Dysidea cinerea
Biemna megalosigma
Ircinia echinata
Holothuria scabra
Petrosia nigricans
Petrosia nigricans
Spongea sp
Cladiella sp
Sarcophyton sp
Asterope sp
Stichopus sp
Thyone bicornis

500
500

2,20
1,85

0,44
0,37

500

4,90

0,98

500
500
500
1000
500

3,00
3,85
8,90
16,60
5,35

0,60
0,77
1,78
1,66
1,07

500

2,60

0,52

500
500
500
1000
500
1000
500
500
500
500

5,20
2,80
2,20
7,70
2,25
10,60
1,15
1,95
2,80
2,35

1,04
0,56
0,44
0,77
0,45
1,06
0,23
0,39
0,56
0,47

500

5,80

1,16

1000

15,50

1,55

1000

11,30

1,13

1000
500
500
500
500
1000
500
500
500
1000
1000

9,30
8,55
1,65
2,10
3,05
12,50
2,80
2,60
1,95
11,90
10,20

0,93
1,71
0,33
0,42
0,61
1,25
0,56
0,52
0,39
1,19
1,02

1000

11,60

1,16

500

5,90

1,18

500

5,85

1,17

1000

10,80

1,08

100
500
1000

1,32
5,85
12,20

1,32
1,17
1,22

Asterina batheri
Archaster typicus
Sarcophyton sp
Sarcophyton sp
30

74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101

KC-74
KC-75
KC-76
KC-77
KC-78
KC-79
KC-80
KC-81
KC-82
KC-83
KC-84
KC-85
KC-86
KC-87
KC-88
KC-89
KC-90
KC-91
KC-92
KC-93
KC-94

San h mm
San h mm
San h mm
San h mm
San h mm
San h mm
San h mm
Hi sm en
Sao bin
Sao bin
Cu gai
Cu gai
C ghim
Sao bin
San h mm
Hi sm en
Hi min cnh xanh
Sao bin
Sao bin
Hi min
Hi min hnh ti

KC-95
KC-96
KC-97
KC-98
KC-99
KC-100
KC-101

Hi min cnh
Hi min xanh
Hi min trng
Sao bin
Sao bin
San h mm
Sao bin rnh nng

Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Sarcophyton sp
Holothuria vagabunda
Astropecten polycanthus
Astropecten polycanthus
Echinothrix calamaris
Echinothrix calamaris
Diadema setosum
Astropecten polycanthus
Sarcaphyton sp
Holothuria vagabunda
Gellius varius
Astropecten polyacanthus
Astropecten monacanthus
Biemna cf. nudelringna
Xestospongea
testudinaria
Gelliodes fibulata
Haliclona clathrata
Mycale plumosa
Astropecten polyacanthus
Astropecten monacanthus
Lobophytum sp
Astropecten polyacanthus

1000
500
500
500
500
500
1000
1000
1000
500
500
500
1000
500
1000
1000
500
1000
500
500

10,80
5,50
7,15
5,00
4,80
6,20
10,20
10,50
10,20
5,55
4,70
4,50
8,50
5,85
12,60
11,80
2,75
11,00
6,95
2,40

1,08
1,10
1,43
1,00
0,96
1,24
1,02
1,05
1,02
1,11
0,94
0,90
0,85
1,17
1,26
1,18
0,55
1,10
1,39
0,48

500

3,00

0,60

500
500
500
1000
1000
500
500

1,60
4,45
5,05
11,30
10,70
6,20
5,85

0,32
0,89
1,01
1,13
1,07
1,24
1,17

II.2. kt qu nh gi hot tnh khng vi sinh vt kim nh (antimicrobial activity)

+ sng lc 101 mu th hot tnh khng vi sinh vt kim nh (cc vi sinh vt
kim nh l cc i din cho cc nhm vi sinh vt gy bnh tiu biu nht thuc cc
nhm vi khun Gr(-); vi khun Gr(+); Nm mc v Nm men). Cc mu c biu hin
dng tnh bc s b ny c pha long theo cc thang nng thp dn, t 5-10
thang nng tnh gi tr nng ti thiu (MIC) m vi sinh vt b c ch pht
trin gn nh hon ton. Ngoi ra, cc mu c biu hin dng tnh cn c tin hnh
chit phn b gia dung mi hu c v nc theo phn cc ca dung mi tng dn t
hexan, sang clorofooc, butanol. Cc dch chit ny c tin hnh xc nh gi tr MIC
nh hng cho cc nghin cu tip theo.
Bng 1: Thng k hot tnh khng VSV kim nh ca 101 mu sinh vt bin
Tng s
mu

101
T l %

Khng 1 Khng 2 Khng 3 Khng 4 Khng 5 Khng 6 Khng 7 Khng 8 S mu c

VSVK VSVK VSVK VSVK VSVK VSVK VSVK VSVK hot tnh

8
7,92

10
9,90

19
18,81

10
9,90

2
1,98

1
0,99

1
0,99

0
0

51
50,49

Kt qu sng lc trn bng 1 cho thy t l cc mu c hot tnh l kh cao: c

51 mu trn tng s 101 mu khng t nht mt chng vi sinh vt kim nh (chim t l
50,49%). Trong , s mu c hot tnh khng 1 vi sinh vt kim nh l 8 mu (chim
7,92 %), khng 2 vi sinh kim nh l 10 mu, khng 3 vi sinh kim nh l 19 mu,
31

khng 4 vi sinh kim nh l 10 mu, khng 5 vi sinh kim nh l 2 mu, khng 6 vi

sinh kim nh 1 mu v khng 7 vi sinh kim nh l 1 mu.
Bng 2: Kh nng khng tng chng VSVK ca 101 mu sinh vt bin
Vi khun Gr (-)
E.coli

S mu c
hot tnh
T l % trn
101 mu th

Vi khun Gr (-)

P.aeruginosa B.subtillis

Nm mc

Nm men

S.aureus

Asp.niger

F.oxysporum

C.albicans

S.cerevisiae

15

31

13

21

10

15

17

18

14,85

30,69

12,87

20,79

9,90

14,85

16,83

17,82

Ngoi ra, nh gi mt cch r rng hn kh nng khng cc chng VSVK

khc nhau, chng ti tin hnh thng k, tnh ton s lng v t l % ca cc mu
sinh vt bin khng tng chng VSVK c th. Kt qu c a ra trong bng 2 v
biu 1:
60
35.00

50

30.69

30.00
25.00
T l %

T l %

40
30
20

20.79
14.85

14.85

12.87

16.83 17.82

9.90

5.00
0.00
E.
co
P.
li
ae
ru
gi
no
sa
B.
su
bt
i ll
is
S.
au
re
us
As
p.
ni
ge
F.
r
ox
ys
po
ru
m
C.
al
bi
ca
ns
S.
ce
re
vi
sia
e

tn
h

SV

h
n

d
n
g

8
g

7
g
K

M
u

h
n

h
n
K

h
n
K

SV

SV

SV

V
g

5
g

4
g
h
n
K

SV

SV

V
3
h
n

h
n

SV

SV

0
h
n

15.00
10.00

10

20.00

Biu 1: thng k hot tnh khng VSV kim nh ca 101 mu sinh vt bin
Kt qu trn cho thy, t l mu sinh vt bin khng cc chng P. aeruginosa v
S. aureus. c bit l vi chng P. aeruginosa, t l mu biu hin hot tnh ln ti
30,69 %. y l trc khun m xanh thuc dng vi khun Gram (-), hiu kh, c th gy
bnh, c bit l nhng ngi c vn v h thng min dch. N l loi in hnh nht
ca chi Pseudomonas v cng l loi vi khun c xc nh l nguyn nhn ch yu
(khong 10%) gy ln cc ca bnh nhim trng c hi c ghi nhn trong cc bnh
vin (hu ht cc ca nhim trng khi b bng l do vi khun ny). Chng nhim vo
nhng ngi b suy gim min dch hay b bng, b thng v nhng ngi phi s dng
ng thng hay my h hp nhn to. Nhim khun Pseudomonas c th dn ti nhim
trng ng tit niu, nhim trng mu, vim phi, vim hng, v nhiu bnh khc.
Pseudononas pht trin thnh khun lc trn phi ca bnh nhn b x nang (cystic
fibrosis - CF), v lm tng tnh trng mn tnh ha ca bnh v phi v tng t l t vong
khi b x nang. Vi khun Pseudomonas thng sinh sng trong t, vng ly li v cc
vng duyn hi. N cng c th sng st di cc iu kin m t c loi vi khun no
chu ng ni, n to ra mt lp mng mng c th chng li s thc bo, v hin nay,
chng khng hu ht cc loi khng sinh ang c s dng. Pseudomonas c th sinh
si c nhng iu kin mi trng rt khc nhau nh: s phng, bn ra chn, thit b
gy m v hi tnh, cc loi nhin liu, my lm m v thm ch c trong nc ct c
lu tr. N cng c pht hin trong my chy thn. Mu xanh c trng ca

32

m l do sc t xanh (pyocyanin) v sc t lc (fluorescein) c sn sinh bi vi khun

Pseudomonas.
Ngoi ra, t l mu c biu hin hot tnh khng t cu khun Staphylococcus aureus
cng t t l 20,79 %. T cu khun c t l gy bnh rt cao, c kh nng gy nhiu
bnh nng cng nh khng khng sinh rt mnh. Cc nh vi khun hc lng danh nh
Robert Koch (1878) v Louis Pasteur (1880) u rt quan tm nghin cu t cu khun
ngay t thi k u ca lch s ngnh vi sinh vt hc. Trn phng din gy bnh, t cu
khun c chia thnh hai nhm chnh: t cu c men coagulase (gm S. aureus v S.
intermedius) v t cu khng c men coagulase. T cu c men coagulase gyln nhiu
bnh l khc nhau nh: cc bnh nhim trng khu tr ti da v nim mc, nhim trng
cc ni c quan, hi chng nhim c, hi chng Thukydides, ng c thc phm.....
Nh vy, vi t l mu th hin hot tnh khng cc chng VSV kim nh (c bit
l khng khng 2 chng vi khun: S. aureus v P. aeruginosa) kh cao ca cc mu sinh
vt bin l mt tin quan trng phc v cho vic tm kim v nghin cu cc hp cht
thin nhin bin theo nh hng hot tnh khng sinh, nhm pht trin cc ch phm
thuc khng sinh th h mi, khc phc tnh trng khng thuc khng sinh nghim trng
hin nay.
II.3. Hot tnh gy c t bo:
ti tin hnh th nghim hot tnh gy c t bo trn ba dng t bo ung
th ngi KB, FL v Hep-G2. Kt qu th nghim 101 mu cho thy c 43 mu c biu
hin hot tnh trong tng s 101 mu th, trong c 12 mu khng mt dng t bo
(chim 11,88 %), 15 mu khng 2 dng t bo (chim 14,85 %) v 16 mu khng c 3
dng t bo (chim 15,84 %).
15.84

34
32.67

33
4.85

32

11.88

T l %

31
42.57

29.7

30
29

27.72

28
Mu c hot tnh

Mu khng 1 dng TB

Mu khng 2 dng TB

Mu khng 3 dng TB

27
26
25
Khng dng t bo
KB

Khng dng t bo
FL

Khng dng t bo
Hep-G2

Biu 2. So snh hot tnh gy c t bo ca cc mu sinh vt bin

Bng 3. Bng tng kt kt qu sng lc hot tnh gy c t bo ca 101 mu

Tng s mu

S mu c hot C hot tnh vi C hot tnh vi C hot tnh vi

tnh
1 dng t bo 2 dng t bo
3 dng t bo

S mu

S mu

S mu

S mu

S mu

101

100

43

42,57

12

11,88

15

14,85

16

15,84

Trong s 101 mu, c 30 mu th hin hot tnh khng dng t bo KB (ung th

biu m), 28 mu khng dng t bo FL (ung th mng t cung) v 33 mu khng dng
t bo Hep-G2 (ung th gan). Kt qu c tng hp trn bng 4 v biu 2.

33

Bng 4: Bng tng kt kh nng cc dng KB ,FL v Hep-G2ca 101 mu

C hot tnh
khng KB

C hot tnh
khng FL

C hot tnh khng

Hep-G2

S mu c hot tnh

30

28

33

T l % trn 101 mu th

29,70

27,72

32,67

T kt qu trn, cc mu c biu hin hot tnh gy c t bo c tin hnh xc

nh gi tr IC50 (nng c ch 50% s pht trin ca t bo). ng thi, cc mu ny
cng c tin hnh chit phn b trong cc dung mi hu c v cc phn on thu
c cng c tin hnh xc nh gi tr IC50. Kt qu chi tit c trnh by trong bo
co chnh. Cc kt qu ny gp phn nh hng cho cc nghin cu tip theo.
II.4. Hot tnh chng xy ho ca cc mu sinh vt bin thng qua phn ng bao vy
gc t do (DPPH).
DPPH c kh nng to ra cc gc t do bn trong dung dch EtOH bo ho. Khi
cho cc cht th nghim vo hn hp ny, nu cht c kh nng lm trung ho hoc bao
vy cc gc t do s lm gim cng hp ph nh sng ca cc gc t do DPPH.
Hot tnh chng xy ho c nh gi thng qua gi tr hp ph nh sng ca dch th
nghim so vi i chng khi c trn my Elisa bc sng 515nm.
Gi tr SC biu hin kh nng bt gi cc gc t do thng qua h xy ho t to
bi DPPH, c tnh theo cng thc c trnh by trong phn phng php.
Bng 5: Tng hp kt qu hot tnh chng xy ho ca cc mu sinh vt bin
Tng s
mu
S mu T l %
101

S mu c
hot tnh
S mu T l %

100

23

22,72

C gi tr SC
t 50-70%
S mu
T l %
18

C gi tr
SC >70%
S mu
T l %

17,82

4,95

120
100

100
77.28

T l %

80
60
40
22.72

20

17.82
4.95

0
Mu dng Mu c SC Mu c SC
tnh
t 50-70%
> 70%

Mu m
tnh

Tng s
mu

Biu 3. So snh t l % cc mu th hin hot tnh chng OXH

Kt qu xc nh gi tr SC cho thy, c 23/101 mu (chim t l 22,72 %) c biu
hin dng tnh vi gi tr SC > 50%. Trong , c 18 mu c kh nng bao vy t 5070% cc gc t do trong th nghim DPPH v 5 mu c kh nng bao gy trn 70% cc
gc t do. Kt qu tng hp c th hin trn bng 5 v biu 3.

34

Cc mu c biu hin dng tnh (SC% > 50 %) c tip tc tin hnh xc nh

gi tr SC50 (gi tr nng mu th c kh nng bao vy 50% cc gc t do), ng thi
tin hnh chit phn b bng cc dung mi hu c. Cc phn on chit ny c tip
tc tin hnh xc nh gi tr SC%. Cc phn on c biu hin dng tnh li tip tc
c xc nh gi tr SC50. Kt qu th nghim c trnh by chi tit trong bo co
chnh. Kt qu trn cho thy s ph hp so vi cc ti liu c cng b. Chng hn
vi cc mu KC-007 (Cladiella sp), KC-010 (Lobophytum sp), KC-014 (Sinularia
abrupta), KC-025 (Sarcophyton mililatensis) c biu hin hot tnh c th c gii
thch bi thnh phn chnh ca cc chi san h mm ny l cc hp cht itecpen dng
cembranoit th hin nhiu hot tnh sinh hc ng quan tm trong c c hot tnh
chng xy ha. y l lp cht c phn cc thp nn s phn b ch yu dch chit
hexan v cloroform, gii thch cho s tp trung hot tnh vo cc dch chit ny ca cc
mu san h mm trn. Cn cc mu KC-056 (Astropecten polyacanthus) v KC-061
(Holothuria scabra) c hot tnh tp trung cc phn on BuOH v nc, c th c
gii thch bi thnh phn hot cht chnh ca cc chi sao bin v hi sm trn u l cc
steroit glycosit cha nhiu n v ng trong phn t, c phn cc rt cao v do
phn b ch yu cc phn on BuOH v nc.

B. nghin cu ha hc theo nh hng hot tnh sinh

hc mt s i tng c la chn
Trn c s cc kt qu nghin cu thu thp, gim nh loi, to dch chit v sng
lc hot tnh sinh hc, kt hp vi cc tiu ch la chn l:
- Hot tnh sinh hc
- Mc kh khn trong cng tc ly mu
- Tr lng thc t ca loi v kh nng c th thu thp mu
- Tra cu cc nghin cu v hot tnh sinh hc v thnh phn ha hc
cng b v loi hoc chi ca i tng nghin cu.
07 i tng ph hp nht c la chn tin hnh thu thp mu lng ln,
to dch chit phc v cho nghin cu ha hc l:
Tn Vit Nam

Tn khoa hc

a im thu mu
Cn o, B Ra Vng tu
Cn C,
Qung Tr

San h mm

Sarcophyton mililatensis

Alcyoniidae

San h mm

Cladiella sp.

Alcyoniidae

Hi sm trng

Holothuria scabra

Holothuridae

Ct B, Hi Phng

Sao bin

Archaster typicus

Archasteridae

Ct B, Hi phng

Hi min xp

Xestospongia testudinaria

Petrosiidae

Hi min cnh
xanh

Gellius varius

Chanlinidae

Hi min

Ianthella sp.

Ianthellidae

35

Qun o Trng
Sa, Khnh Ha
H Long, Qung
Ninh
Qun o Trng
Sa, Khnh Ha

Chng III. Nghin cu ho hc theo nh hng hot tnh

sinh hc ca mt s loi san h mm
III.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi
san h mm sarcophyton mililatensis (ALcyoniidae)

- Tn Vit Nam: San h mm

- Tn khoa hc: Sarcophyton
mililatensis
- Thi gian thu mu: thng 6
nm 2007
- a im thu mu: hn By
Cnh, huyn o Cn o, tnh
B Ra - Vng Tu
- Ngi nh loi: TS. Cng
Thung v cc cng s, Vin Ti
nguyn v Mi trng Bin,
VAST

Phn lp v tinh ch cc hp cht:

San h mm Sarcophyton
mililatensis ti (20 kg)
- Chit MeOH, ct loi dung mi di p sut gim
- Loi mui bng cch chy qua ct Dianion vi
dung mi ra gii l nc ct, sau thu hi cht
bng MeOH.

B sung nc

SM - Cn chit MeOH

B sung cloroform
cloroform: nc 1/1

Lp nc
B sung BuOH

C - Cn cloroform (150 g)

BuOH: nc 1/1

B- Cn BuOH (20 g)

Lp nc

N - Cn nc (30 g)

B sung MeOH
B sung hexan

MeOH: hexan 1/1

M - Cn tan trong
MeOH (50 g)

H - Cn tan trong
hexan (100 g)

S III.1a: Chit phn on mu san h mm

36

Mu san h mm ti (20 kg) c xay nh v chit 3 ln bng MeOH nng

(50oC mi ln 3h) v ct loi dung mi di p sut gim thu c 200 g cn dch
MeOH. Cn ny c ha vo nc ct v chit phn on ln lt vi cc dung mi
cloroform v n-butanol. Cn dch cloroform (150 g) c ha vo metanol v chit vi
hexan thu c cc cn dch: cn tan trong hexan (100 g) v tan trong metanol (50 g)
(s III.1a). Cn tan trong hexan c tin hnh cha ct th trn sc k ct pha
thng ( = 10 cm, L = 50 cm) gradient nng axtn trong hexan t 0 n 100% thu
c 5 phn on nh hn k hiu l H1 - H5. Phn on H2 (70g) c tin hnh trn
sc k ct pha o vi h dung mi ra gii l MeOH/nc 4/1 thu c cc hp cht
SM6 (50 g) v SM7 (0,5 g) (s III.1.b).
Cn H - (100 g)
Silica gel CC
Hexan/Axeton 100/0-0/100

H1 (5 g)

H2 (70 g)

H3 (9 g)

H4 (11 g)

H5 (5 g)

Silica gel CC pha o

MeOH/H2O 4/1

SM-6
(50 g)

H4A
(1 g)

Silica gel CC pha thng

n-hexan/axeton 7/1

SM-7
(0,5 g)

H4B
(3 g)

H4C
(2 g)

H4D
(1 g)

Silica gel CC pha o

MeOH/H2O 5/1

SM1
(40 mg)

SM4
(20 mg)

SM5
(9 mg)

H4E
(4 g)
Silica gel CC
H/C/M 10/1/0.5

SM2
(15 mg)

SM3
(20 mg)

S III.1.b: Phn lp v tinh ch cc hp cht t cn hexan

Phn on H4 (11 g) c tip tc phn tch trn sc k ct pha thng vi h
dung mi ra gii l hexan/axeton 7/1 thu c 5 phn on nh hn na k hiu l
H4A n H4E. T phn on nh H4B (3g) cc hp cht SM1 (40 mg), SM4 (20 mg)
v SM5 (9 mg) c lm sch bng phng php sc k ct silica gel pha o vi h
dung mi ra gii l MeOH/nc 5/1. Phn on H4D (1 g) uc tin hnh phn lp trn
sc k ct pha thng vi h dung mi ra gii hexan/cloroform/MeOH 10/1/0.5 thu
c cc hp cht SM2 (15 mg) v SM3 (20 mg) (s III.1.c).
37

Cn M - (50 g)

Silica gel CC
Cloroform/MeOH 100/0-0/100

M1 (20g)

M2 (2 g)

Silica gel CC pha

o MeOH/H2O: 5/1

M3 (5 g)

M4 (10 G)

M5 (13 g)

Silica gel CC pha

o MeOH/H2O: 5/1

SM8
(20 mg)
SM9
(200 mg)

SM10
(100 mg)

S III.1.c. Phn lp v tinh ch cc hp cht t cn chit M

Xc nh cu trc ha hc v hot tnh sinh hc ca cc hp cht:
Cu trc ha hc ca cc hp cht c xc nh bng cch phn tch chi tit cc
d kin v hng s vt l, ph cng hng t ht nhn, ph khi lng v so snh cc s
liu vi cc ti liu c cng b. Do khun kh ca bo co tm tt c hn
chng ti ch chnh by y qu trnh xc nh cu trc ca mt hp cht in hnh
SM1:
Hp cht SM1 c phn lp di dng cht du khng mu. Cng thc phn t
ca n c xc nh l C21H34O3 bng cc d kin ph khi lng (positive ESI-MS
m/z: 357 [M+Na]+, 317 [M-H2O+H]+, 303 [M-MeOH+H]+, 285 [M-MeOH-H2O+H]+) v
ph cng hng t ht nhn (Bng III.1).

Hnh III.1.a. Ph 1H-NMR ca SM1

38

18

4
7

HO

16

21

OMe

19

15

17

14

11
20

Hnh III.1.b. Cu trc ha hc ca SM1

1

Ph H-NMR ca SM1 xut hin cc tn hiu c trng ca hai proton olefin cng
hng ti 5,22 (1H, d, J = 11,0 Hz, H-3) v 5,36 (1H, d, J = 11,0 Hz, H-11); hai
proton oximetin ti 5,38 (1H, d, J = 11,0 Hz, H-2) v 3,38 (1H, t, J = 7,0 Hz, H-7);
cc proton ca nhm oximetilen ti 4,45 (1H, d, J = 11,0 Hz, Ha-16)/4,56 (1H, dd, J =
11,0 v 4,0 Hz, Hb-16); cc proton ca 4 nhm metyl bc bn ti 1,69 (3H, s, H-17),
1,81 (3H, s, H-18), 1,09 (3H, s, H-19) v 1,67 (3H, s, H-20); mt nhm metoxy ti
3,19 (3H, s, H-21) v cc tn hiu proton ca cc 6 nhm metilen (Bng III.1).

Hnh III.1.c. Ph 13C-NMR v cc ph DEPT ca SM1

39

Bng III.1. Kt qu ph NMR ca SM1

C

C *

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

38,8
28,3
73,7
79,3
31,5
21,3
127,9
135,7
36,8
26,9
-

133,97
85,80
124,47
142,60
38,66
29,01
74,03
79,57
31,97
21,63
126,38
136,54
38,21
24,41
128,00
78,46

17
18
19
20
21

9,6
16,7
18,1
16,2
48,9

10,67
16,49
18,49
16,08
48,96

Ha, c
multiplicity (J in Hz)
5,36 d (11,0)
5,22 d (11,0)
2,22 m/2,18 m
1,77 m/1,55 m
3,38 t (7,0)
1,70 m/1,65 m
2,15 m/1,99 m
5,36 t (7,0)
2,29 m/2,08 m
2,39 m/1,64 m
4,56 dd (11,0, 4,0)
4,45 d (11,0)
1,69 s
1,81 s
1,09 s
1,68 s
3,19 s

HMBC
(HC)

5, 18

5, 6, 8, 19
10
10, 13, 20
1, 11, 12, 14
12

1, 15, 16
3, 4, 5
7, 8, 9
11, 12, 13
8

o trong CDCl3, b125 MHz, c500 MHz, *s liu ph ca hp cht (-)-7-Hydroxy-8-methoxydeepoxysarcophine (Kobayashi M., and Hirase T., Chem. Pharm. Bull., 1990, 38(9), 2442-2445)

Hnh III.1.d. Ph HSQC ca SM1

Trn ph 13C-NMR ca SM1 xut hin 20 tn hiu c trng ca mt hp cht
ditecpen dng cembranoit. Trong , cc tn hiu cng hng ti 124,47 (CH, C40

3)/142,60 (C, C-4), 126,38 (CH, C-11)/136,54 (C, C-12) v 133,97 (C, C-1)/128,0 (C, C15) khng nh s tn ti ca hai lin kt i b th 3 v tr v mt lin kt i b th
hon ton. Bn nhm metyl bc ba v mt nhm metoxy c khng nh tng ng bi
cc tn hiu cng hng ti 10,67 (C-17), 16,49 (C-18), 18,49 (C-19), 16,08 (C-20) v
48,96 (C-21). Ngoi ra cc tn hiu c trng cho hai nhm oximetin, mt nhm
oximetilen v mt nhm ete bc bn cng c khng nh tng ng ti 85,80 (C-2),
74,03 (C-7), 78,46 (C-16) v 79,57 (C-8). Cc tn hiu cacbon c gn vi cc tn hiu
proton tng ng thng qua ph cng hng t ht nhn hai chiu tng tc trc tip
HSQC (Bng III.1 v Hnh III.1.d).

Hnh III.1.e. Ph HMBC ca SM1

18

HO

H
O

16

21

OMe

19

15

11
17

14

HMBC
20

COSY

Hnh III.1.f. Cc tng tc HMBC v COSY chnh ca SM1

Trn ph 1H-1H COSY xut hin cc tng tc gia H-2/H-3, H2-5/H2-6/H-7, H29/H2-10/H-11 v H2-13/H2-14 (Hnh III.1.f). Kt qu trn, cng vi cc tn hiu tng
tc HMBC gia H-2/C-1, H3-18/C-3, H3-18/C-4, H3-18/C-5, H3-19/C-7, H3-19/C-8, H319/C-9, H3-20/C-11, H3-20/C-12, H3-20/C-13 v H2-13/C-1 cho php ghp ni cc phn
cu trc t C-1 n C-14 ca vng 14 cacbon.

41

Hnh III.1.g. Ph 1H-1H COSY ca SM1

Hnh III.1.h. Ph ROESY ca SM1

42

V tr ca nhm metyl C-17 c khng nh bi cc tn hiu tng tc t cc

proton ca n ti C-1, C-15 v C-16 nhn c trn ph HMBC. Cng vi cc phn tch
nu phn trn cho php khng nh ton b cu trc phng ca SM1 nh a ra
Hnh III.1.b.
Cu hnh ti cc v tr bt i ca SM1 c xc nh bng cch so snh s liu
ph NMR ca n vi cc hp cht c cu trc tng t c cng b (Kobayashi M.,
and Hirase T., Chem. Pharm. Bull., 1990, 38(9), 2442-2445 v Daniela, G., Hesham, S. M. S., Kamel, H.
S., Mohamed, H., and Karlheinz, S., Natural Product Research, 2006, 20, 285-291) v c khng

nh thm bng cc tng tc trn ph ROESY. Cu hnh ca cc lin kt i C-3/C-4

v C-11/C-12 c khng nh u dng trans da trn s liu chuyn dch ha hc
13
C ca cc cacbon C-18 v C-20 tng ng ti 16,49 v 16,08 (nh hn 20 ppm).
CH3

H
O

HO

H
H

CH3

H 3C
CH3
OCH3

Hnh III.1.i. Cu hnh khng gian v cc tng tc

ROESY then cht ca SM1
Trn ph ROESY ca SM1, tn hiu tng tc cho gia proton H-2 vi cc
proton H3-18 cho php khng nh chng u nm v tr v cu hnh ca lin kt i
C-3/C-4 dng trans. Cc tn hiu tng tc cho ROESY gia H-3 v H-7, H-3 v Ha-6
( 1.77), Ha-6 v H3-21 cho thy H-3, Ha-6 v H3-21 u nm v tr . T tt c cc
phn tch nu trn, cu trc ha hc ca SM1 c xc nh cho mt hp cht mi l
(-)-7-hydroxy-8-methoxydeepoxy-sarcophytoxide.

Hnh III.1.j. Ph ESI-MS ca SM1

43

Bng cc phng php tng t, chng ti xc nh c cu trc ha hc ca

ton b cc cht c phn lp t loi san h mm Sarcophyton mililatensis. Cc hp
ny c tin hnh nh gi hot tnh sinh hc theo cc phng php c trnh by
phn phng php. Kt qu chi tit c tm tt sau y:
1. Hp cht SM1: (-)-7-hydroxy-8-methoxydeepoxysarcophytoxide (Cht mi, ln
u tin c phn lp t thin nhin)
- Cu trc ha hc:
- Hot tnh sinh hc:
18
+Hot tnh chng long xng
H

4
7

HO

16

21

Hot lc ca
enzym ALP
(%)

Hm lng
collagen
(%)

S khong ha
(%)

0,3

127,81,392

108,21,860

110,62,470

125,12,760

101,92,680

106,70,403

30

115,56,288

91,483,231

105,12,619

OMe

19

15

17

14

11

+ Hot tnh gy c t bo
20
- Cht du khng mu
Dng KB: IC50 = 3,23 g/ml
- Cng thc phn t: C21H34O3, M = 334
Dng FL: IC50 = 2,77 g/ml
- Ph khi lng: m/z 357 [M+Na]+ 317 Dng Hep-G2: IC50 = 3,45 g/ml
[M-H2O+H]+, 303 [M-MeOH+H]+, 285
[M-MeOH-H2O+H]+
2. Hp cht SM2: (-)-17-hydroxysarcophytonin A (Cht mi, ln u tin c phn
lp t thin nhin)
- Cu trc ha hc:
- Hot tnh sinh hc:
+Hot tnh chng long xng
H

OH

Hot lc ca
enzym ALP
(%)

Hm lng
collagen (%)

S khong
ha (%)

0,3

131,06,834

101,03,154

97,111,485

127,21,292

102,51,832

101,72,556

30

130,83,774

111,41,208

107,93,767

+ Hot tnh gy c t bo
Dng FL: IC50 = 3,06 g/ml
Dng Hep-G2: IC50 = 3,69 g/ml

- Cht du khng mu
- Cng thc phn t: C20H30O2, M = 302
- Ph khi lng: m/z 325 [M+Na]+, 303
[M+H]+, 285 [M-H2O+H]+

3. Hp cht SM3: sarcophytol V (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
- Ph khi lng: m/z: 327 [M+Na]+, 287 [MH2O+H]+, 269 [M-2H2O+H]+
- Hot tnh sinh hc:
OH
+Hot tnh chng long xng
OH

- Cht du khng mu
- Cng thc phn t: C20H32O2, M =
304

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

127,02,449
121,02,237
113,22,973

102,70,137
101,01,938
110,41,440

96,771,650
97,721,546
101,60,990

+ Hot tnh gy c t bo
Dng KB: IC50 = 3,77 g/ml
44

4. Hp cht SM4: sarcophytol X (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
- Ph khi lng: m/z 334,8 [M-H2O+H]+,
316,8 [M-2H2O+H]+, 298,8 [M-3H2O+H]+)
O
- Hot tnh sinh hc:
OH
Chng xy ha trn h DDPH vi gi tr
O
OH
ED50 = 75,5 g/ml
OH

- Cht du khng mu
- Cng thc phn t: C20H32O5, M =
352
5. Hp cht SM5: (+)-7,8-dihydroxydeepoxysarcophytoxide (Cht mi, ln u tin
c phn lp t thin nhin)
- Cu trc ha hc:
-Hot tnh sinh hc:
H
+Hot tnh chng long xng
O
Hot lc ca
Hm lng
S khong
HO
M
enzym ALP (%)
collagen (%)
ha (%)
HO
0,3
128,92,660 109,51,898 100,71,684
3
30

- Cht du khng mu
- Cng thc phn t: C20H32O3, M =
320
- Ph khi lng: m/z 343 [M+Na]+,
303 [M-H2O+H]+

139,52,237
123,41,814

106,84,880
99,794,012

104,01,277
105,42,868

+ Hot tnh gy c t bo
Dng KB: IC50 = 4,23 g/ml
Dng FL: IC50 = 1,97 g/ml
Dng Hep-G2: IC50 = 3,95 g/ml

6. Hp cht SM6: Hp cht (+)-sarcophine

- Cu trc ha hc:
- Hot tnh sinh hc:
H
+Hot tnh chng long xng
O

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

133,32,091
130,26,923
121,91,062

103,50,836
109,72,000
104,01,716

96,171,540
99,870,941
99,661,515

+ Hot tnh gy c t bo
Dng FL: IC50 = 4,21 g/ml
- Cht du khng mu
Dng Hep-G2: IC50 = 3,55 g/ml
- Cng thc phn t: C20H32O3, M =
+
Hot
tnh chng xy ha
320
+
ED
50 = 62,3 g/ml
- Ph khi lng: m/z 339 [M+Na] ,
+
+
317 [M+H] , 299 [M-H2O+H] .

45

7. Hp cht SM7: sarcophytoxide

- Cu trc ha hc:
H

- Hot tnh sinh hc:

+Hot tnh chng long xng

Hot lc ca
enzym ALP (%)

Hm lng
collagen (%)

S khong
ha (%)

0,3
3
30

127,82,995
125,13,296
117,42,995

103,23,421
106,91,636
109,11,734

96,102,443
97,783,635
102,81,296

+ Hot tnh gy c t bo
Dng FL: IC50 = 4,32 g/ml

- Cht du khng mu
- Cng thc phn t: C20H30O2, M =
302
- Ph khi lng: m/z: 325 [M+Na]+,
303 [M+H]+, 285 [M-H2O+H]+.

8. Hp cht SM8: sarcomilasterol (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
- Hot tnh sinh hc:
+Hot tnh chng long xng
28

21

26

24

18

17

11

HOH2C

27

13

19

25

20

14

1
8

10
3

5
6

HO

OH

- Tinh th hnh kim mu trng: mp 160161oC

Hot lc ca
enzym ALP
(%)

Hm lng
collagen (%)

S khong ha
(%)

0,3
3
30

119,15,214
118,01,486
86,410,212

106,51,822
104,01,254
69,10,495

101,11,8564
107,21,680
89,383,609

+ Hot tnh gy c t bo
Dng KB: IC50 = 2,23 g/ml
Dng FL: IC50 = 3,57 g/ml
Dng Hep-G2: IC50 = 2,65 g/ml

- Cng thc phn t: C28H48O4, M = 448

- Ph khi lng: m/z 471,1 [M+Na]+,
449,1 [M+H]+, 431,1 [M-H2O+H]+

9. Hp cht SM9: sarcoaldesterol B

- Cu trc ha hc:
28

21

26
24

18

HO
19

17
13

11

25

20

27

9
10

- Ph khi lng: m/z: 473,1 [M+Na]+, 433,1

[M-H2O+H]+, 415,1 [M-2H2O+H]+, 397,1 [M3H2O+H]+
- Hot tnh sinh hc:
+Hot tnh chng long xng

HO
OH
OH

0,3
3
30

- Cht bt mu trng
- Cng thc phn t: C28H50O4, M =
450
46

Hot lc ca
enzym ALP
(%)
126,31,486
124,80,368
88,542,872

Hm lng
collagen (%)
109,72,302
108,01,012
98,001,801

S khong
ha (%)
100,91,763
104,51,808
97,851,195

10. Hp cht SM10: ergosta-1,3,5,6-tetraol.

- Cu trc ha hc:
- Ph khi lng: m/z 473,1 [M + Na]+, 450,9 [M
28

21

26
24

18

20
17

OH
19

13

11

25

27

+H]+

- Hot tnh sinh hc:

+Hot tnh chng long xng

10
3

Hot lc ca
enzym ALP (%)

0,3
3
30

132,96,623
126,54,246
91,300,766

HO
OH

Hm lng
collagen (%)

S khong ha
(%)

105,61,259
100,03,690
89,634,532

104,501,324
98,123,197
105,82,156

OH

- Tinh th hnh kim mu trng

- Cng thc phn t: C28H50O4, M = 450
T loi san h mm Sarcophyton mililatensis, 10 hp cht (6 cht mi) c
phn lp v xc nh cu trc, trong c 07 hp cht ditecpen dng cembranoit (5 cht
mi), 01 hp cht 9,11-scosterol mi v 02 hp cht steroit mang nhiu nhm hydroxy
trong phn t. Hot tnh gy c t vo, chng xy ha v chng long xng ca cc
hp cht c nh gi.
Trong qu trnh hnh thnh xng, cc nguyn bo xng l t bo ct li trong
qu trnh hnh thnh cht c bn ca xng (bone matrix) v s vi ha. S to xng
bt u bi s sn sinh v tit collagen loi I ca cc nguyn bo xng, thnh phn to
nn 90% cht c bn ca xng, hay m tin ct. Cc nguyn bo xng c tng hm
lng enzym ALP (enzym ct nhm phosphat). Enzym ALP c gii phng vo m
tin ct khi ng qu trnh tch t cc nguyn t khong. Sau qu trnh khong ha,
xng hon chnh ngy cng tr ln cng v chc vi cc c tnh c hc cn thit
chng li cc ngoi lc, nng c th v bo v cc ni c quan. Cc sterol v
cembranoit ditecpen th hin hot tnh kch thch mnh m s bit ha nguyn bo
xng. c bit, hp cht SM1 v SM10 tng cng mnh m c hot lc enzym ALP,
s tng hp collagen v s tch t canxi ca t bo MC3T3-E1. Theo cc hp cht
ny c th ng vai tr quan trng trong vic hnh thnh xng
Cc hp cht SM1, SM5 v SM8 th hin hot tnh trn c 3 dng t bo th
nghim. Hp cht SM2 v SM6 th hin hot tnh 2/3 dng t bo th nghim. Hp
cht SM3 v SM7 th hin hot tnh 1/3 dng t bo th nghim. Theo cc ti liu
cng b, (+)-sarcophine (SM6) l mt trong nhng thnh phn quan trng trong h
thng phng v ha hc ca cc loi san h mm chng li k th. Ngoi ra, n cn th
hin hot tnh gy c t bo cao i vi nhiu dng t bo nh dng A549 (ED50 =
24,66 g/ml), HT-29 (ED50 = 22,88 g/ml) v c bit vi dng P-388 (ED50 = 3,27
g/ml) (Duh, C. Y., and Hou, R. S., J. Nat. Prod., 1996, 59, 595-598). Khng nhng th (+)sarcophine cn th hin kh nng chng s di tr ca cc t bo ung th rt cao [s di
tr (migration) v tng sinh (proliferation) l cc yu t to nn qu trnh di cn
(metastasis) ca bnh ung th], kh nng ny khng phi do tnh c t bo to nn m
l do n c ch cc c ch di tr ca cc t bo ung th. y l mt du hiu tt, t c
s cho cc nghin cu ton din hn v hot tnh khng ung th ca cc hp cht
cembranoit.
47

Ch c hp cht SM4 v SM6 th hin hot tnh chng xy ha vi gi tr ED50

tng ng l 75,5 v 62,3 g/ml. iu ny cho thy s ph hp v theo ti liu cng
b, (+)-sarcophine (SM6) th hin hot tnh chng xy ha, c nh gi bng kh
nng c ch s sn sinh ion m peoxit (O2-) v thromboxane B2 (TXB2) bi cc t bo
tiu thn kinh m no chut s sinh c hot ha vi lipopolysaccharit ca E.
coli (LPS) (Sawant, S., Youssef, D., Mayer, A., Sylvester, P., Wali, V., Arant, M., and Sayed, K. E.,.
Chem. Pharm. Bull., 2006, 54, 1119-1123) khi c kch thch bng phorbol 12-myristate 13acetate (PMA). Hot tnh chng xy ha ca SM4 v SM6 c th c gii thch bi s
c mt ca nhm ktn trong phn t. SM6 c phn lp vi hm lng kh cao, iu
ny gi m cho nhng nghin cu ng dng tip theo ca hp cht ny theo nh hng
hot tnh chng xy ha.
Cc kt qu c ng trn 04 cng trnh trong c 02 cng trnh trn tp ch chuyn ngnh
quc t v 02 bi trn tp ch chuyn ngnh quc gia. Di y l danh sch cc cng trnh cng b:
1.

2.

3.

4.

Chau Van Minh, Nguyen Xuan Cuong, Tran Anh Tuan, Eun Mi Choi, Young Ho Kim, and Phan Van
Kiem, A new 9,11-secosterol from the Vietnamese Sea soft coral Sarcophyton mililatensis, increases
the function of osteoblastic MC3T3-E1 cells. Natural Product Communications (USA), 2007, 2(11),
1095-1100.
Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van Kiem, Chau Van Minh, Eun Mi Choi, and Young
Ho Kim, New Cembranoid Diterpenes from the Vietnamese Soft Coral Sarcophyton mililatensis
Stimulate Osteoblastic Differentiation in MC3T3-E1 Cells. Chemical and Pharmaceutical Bulletin
(Japan), 2008, 56(7), 988-992.
Chau Van Minh, Nguyen Xuan Cuong, Tran Anh Tuan, Eun Mi Choi, Young Ho Kim, and Phan Van
Kiem, Polyhydroxylated sterols from the Vietnamese Sea soft coral Sarcophyton mililatensis,
increases the function of osteoblastic MC3T3-E1 cells. Tp ch Khoa hc (i hc S phm H Ni
II), 2007, 1(1), 49-54.
Nguyen Xuan Cuong, Tran Anh Tuan, Phan Van Kiem, and Chau Van Minh, Sarcophytol X, a new
cembranoid diterpene from the vietnamese sea soft coral Sarcophytol mililatensis. Tp ch Khoa hc
(i hc S phm H Ni II), 2007, 1(1), 55-60.

Ngoi ra, cc kt qu trn c s dng cho mt lun n Thc s bo v thnh cng lun n vi ti:
Nghin cu thnh phn ha hc v hot tnh sinh hc ca loi san h mm Sarcophyton
mililatensis (ThS. Trn Anh Tun)

48

III.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi

san h mm cladiella sp. (ALcyoniidae)

- Tn Vit Nam: San h

mm
- Tn khoa hc: Cladiella sp
- Thi gian thu mu: thng
11 nm 2006 v thng 3
nm 2007
- a im thu mu: Cn
C, tnh Qung Tr
- Ngi nh loi: TS.
Cng Thung v cc cng s,
Vin Ti nguyn v Mi
trng Bin, VAST

Phn lp v tinh ch cc hp cht:

San h mm Cladiella sp. ti
(2 kg)
- Chit MeOH, ct loi dung mi di p sut
gim
- Loi mui bng cch chy qua ct Dianion vi
dung mi ra gii l nc ct, sau thu hi
cht bng MeOH.
B sung nc
SM - Cn chit MeOH (20g)
B sung n-hexan
n-hexan: nc 1/1

Lp nc
B sung CHCl3

SH - Cn n-hexan (5 g)

CHCl3: nc 1/1

Lp nc
B sung BuOH

SC - Cn CHCl3 (7 g)

BuOH: nc 1/1
lp nc

SB - Cn BuOH (4 g)

SN - Cn nc (4 g)

S III.2.a. S chit phn on mu san h mm

49

Mu san h mm ti (2 kg) c xay nh, sau chit vi metanol ba ln, mi

ln 72 gi. Cc dch chit metanol c gp li v ct loi dung mi di p sut gim
thu c cn chit MeOH th. Cn chit ny c ha vo nc ri tin hnh loi mui
trn ct Dianion vi dung mi ra gii l nc. Dng MeOH ra gii thu hi cht thu
c 20 gam dch c. Dch c ny c ho tan vo 01 lt nc ct sau c chit ln
lt bng n-hexan, cloroform, v BuOH (mi ln 02 lit) thu c cc cn dch tng
ng: hexan (5 g), cloroform (7 g), BuOH (4 g) v nc (4 g) (s III.2.a).
SH - (5 g)
Silica gel CC
Hexan/Axeton 100/0-0/100

SH1 (3000mg)

SH2 (400mg)

SH3 (250mg)

SH4 (750mg)

Silica gel CC
n-hexan/Etylaxetat 10/1

SH-8
(200mg)

SH5 (600g)

Silica gel CC
n-hexan/axeton 7/1

SH-7
(100mg)

S III.2.b. S phn lp cc hp cht t cn chit n-hexan

SC (7 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

SC1 (2g)

SC2 (1,2g)

SC3 (1,3g)

Silica gel CC
n-hexan/axeton
3/2

Silica gel CC
Cloroform/axeton: 10/1

SH-5
(500mg)

SC4 (1,5g)

SH-6
(70mg)

SH-4
(8mg)

SC5 (1g)

Silica gel CC
CHCl3/MeOH
7/1
SH-3
(9mg)

S III.2.c. S phn lp cc hp cht t cn chit cloroform

Cn chit n-hexan: SH (5 g) c tin hnh phn tch trn sc k ct lp li nhiu
ln vi cc h dung mi thch hp thu c cc hp cht SH-7 v SH-8. Cn chit
clorofrom: SC (7 g) c tin hnh chy ct th trn sc k ct pha thng gradient
nng metanol trong cloroform thu c 5 phn on nh hn l SC1-5. Cc phn
on SC1, SC4 v SC5 c tip tc phn tch trn sc k ct pha thng vi h dung
mi ra gii thch hp thu c cc hp cht SH-3, SH-4, SH-5 v SH-6. Bng cc
phng php tng t, cc hp cht SH-1 v SH-2 c phn lp t cn chit BuOH
50

SB - (4 g)
Silica gel CC
Cloroform/MeOH/H2O: 5/1/0 - 2/1/0.2

SB1 (1.0 g)

SB2 (2.5 g)

SB3 (1.5 g)

Silica gel CC
CHCl3/MeOH
7/1
SH-1
(4mg)

SB4 (600mg)

SB5 (1.4 g)

Silica gel CC
Etylaxetat/CHCl3/MeOH
6/2/1
SH-2
(12mg)

S III.2.d. S phn lp cc hp cht t cn chit butanol

Xc nh cu trc v hot tnh sinh hc ca cc hp cht:
S dng kt hp cc phng php ph hin i, chng ti xc nh c cu
trc ha hc ca ton b cc cht c phn lp t loi san h mm Cladiella sp. Cc
hp cht ny c tin hnh nh gi hot tnh sinh hc theo cc phng php c
trnh by phn phng php. Kt qu chi tit c tm tt sau y:
1. Hp cht SH1: cladiasterol (Cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:
- Ph khi lng: m/z 615,2
[M+Na]+, 454,2 [MHO
fucose+Na]+, 591,4 [M-H]- Ph khi lng phn gii cao:
m/z 615,389 [M+Na]+ (tnh ton
O H
OH
l thuyt cho C34H56O8Na:
615.3873)
O
H
- Hot tnh sinh hc:
HO
O
+ Hot tnh gy c t bo
Dng KB: IC50 = 4,43 g/ml
OH
HO
+ Hot tnh chng xy ha
ED50 = 77,8 g/ml
- Cht bt mu trng
- Cng thc phn t: C34H56O8, M = 592

51

2. Hp cht SH2: cladiaceramide (Cht mi, ln u tin c phn lp t thin

nhin)
- Cu trc ha hc:
- Ph khi lng phn gii cao: m/z
OH
574,489 [M+Na]+ (tnh ton l
O
thuyt cho C34H65NO4Na:
1'
3'
16'
10
574,4811)
HN
- Hot tnh sinh hc:
+ hot tnh khng NF-B
18
HO
8
1
4
IC50 = 14,8 1,73 M
7
+ Hot tnh gy c t bo
OH
Dng KB: IC50 = 4,43 g/ml
- Cht bt v nh hnh mu trng
- Cng thc phn t: C34H65NO4 (M = 551)
- Ph khi lng: m/z 574,4 [M+Na]+, 552,4
[M+H]+, 534,4 [M-H2O+H]+, 550,4 [M-H]3. Hp cht SH3: cladiacerebroside (Cht mi, ln u tin c phn lp t thin
nhin)
- Cu trc ha hc:
- Ph khi lng: m/z: 848,6
[M+H]+, 846,8 [M-H]OH
Cladiella sp.
O
- Hot tnh sinh hc:
16
+ Hot tnh gy c t bo
HN
OH
Dng KB: IC50 = 3,77
O
HO
O
g/ml
7
HO
SO3H
Dng Hep-G2: IC50 = 4,72
HO
g/ml
- Cht bt v nh hnh mu trng
+ Hot tnh chng xy ha
- Cng thc phn t: C45H85NO11S, M = 847
ED50 = 65,5 g/ml
1'

4"

20'

6"

18

5"

1"

2"

3"

19

4. Hp cht SH4: 3,6,11-trihydroxy-24-methylene-9,11-secocholest-7-en-9-one

- Cu trc ha hc:
28
- Ph khi lng: m/z 447,2 [M + H]+,
26
445,4 [M-H]21
HO
24
23
25
22
20
- Hot tnh sinh hc:
11
18
+ Hot tnh khng NF-B:
12
27
17
IC50 = 32,5 1,80 M
13
19
O
16
+
Hot
tnh gy c t bo
14
15
9
1
Dng KB: IC50 = 4,23 g/ml
8
10
2
5
Dng FL: IC50 = 2,97 g/ml
3
7
6
4
HO
Dng Hep-G2: IC50 = 1,95 g/ml
H

OH

- Tinh th hnh kim mu trng: mp 162-164oC

- Cng thc phn t: C28H46O4, M = 446

52

5. Hp cht SH5: cholesterol

- Cu trc ha hc:
21

21

20

24

- Tinh th hnh kim mu trng: mp 148149oC

- Cng thc phn t: C27H46O, M=386

25

18
17

26

19
1
8
3
5

HO

6. Hp cht SH6: batyl alcohol (batilol)

- Cu trc ha hc:
1

- Tinh th mu trng: mp 70-71oC

- Cng thc phn t: C21H44O3, M = 344
- Ph khi lng: m/z 345,1 [M+H]+

OH

HO

1'

18'

CH2(CH2)16CH3

7. Hp cht SH7: chimyl alcohol

- Cu trc ha hc:
1

- Tinh th mu trng: mp 64-65oC

- Cng thc phn t: C19H40O3, M=316
- Ph khi lng: m/z: 317,1 [M+H]+

OH

HO

1'

16'

CH2(CH2)14CH3

8. Hp cht SH8: 1-heptadecanol

- Cu trc ha hc:
1

16
17

HO

14

CH3

- Tinh th mu trng: mp 59,5-60.0oC

- Cng thc phn t: C17H36O, M=256
- Ph khi lng: m/z 279,0 [M+Na]+,
255,0 [M-H]-, 237,0 [M-H2O-H]-

Kt qu nghin cu nh gi hot tnh cho thy, ch c hai hp cht l SH2 v

SH4 th hin hot tnh khng NF-B. iu ny cho thy s ph hp so vi cc ti liu
cng b v SH2 l mt hp cht ceramit cn SH4 l mt 9,11-secosterol, hai lp cht
th hin rt nhiu hot tnh qu, cn c quan tm nghin cu pht trin thnh cc
ch phm thuc c gi tr cao, phc v cng tc chm sc sc khe cng ng. Hp cht
SH1 cng l mt 9,11-secosterol tuy nhin n li dng dn xut glycosit, v mt iu
th v l y l dn xut glycosit u tin ca cc hp cht 9,11-secosteroit c phn
lp t thin nhin. Hp cht ny khng th hin hot tnh khng NF-B c th l do s
nh hng ca n v ng fucose trong phn t.
Ngoi ra c 4 hp cht th hin hot tnh gy c t bo. Trong hp cht SH1
ch c hot tnh gy c t bo i vi dng KB vi IC50 = 4,43 g/ml, SH2 v SH4 th
hin hot tnh trn c 3 dng t bo th nghim, SH3 th hin hot tnh trn 2 dng t
bo l KB v Hep-G2 vi gi tr IC50 = 3,77 v 4,72 g/ml. Mt iu ng ch l hp
cht SH2 l mt hp cht mi, th hin hot tnh gy c t bo trn c 3 dng t bo
th nghim v c phn lp vi hm lng tng i cao t loi san h mm Cladiella
53

sp. iu ny gi m cho cc nghin cu tip theo ca hp cht ny theo nh hng hot

tnh gy c t bo nhm pht trin cc loi dc liu mi c tc dng phng v iu tr
bnh ung th.
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc loi san h mm
Cladiella sp, chng ti bin son v ng ti 01 cng trnh trn tp ch chuyn ngnh quc t l:
Chau Van Minh, Phan Van Kiem, and Nguyen Xuan Cuong, New 9,11-secosterol glycoside and ceramide from a soft coral
Cladiella sp. living in Vietnamese sea. ASEAN Journal on Science and Technology for Development, 2007, 24(3), 275-283.

Chng IV. Nghin cu ho hc theo nh hng hot tnh

sinh hc ca mt s loi da gai.
IV.1. nghin cu ha hc theo nh hng hot tnh sinh hc
loi hi sm holothuria scabra (holothuridae)

- Tn Vit Nam: hi sm trng

- Tn khoa hc: Holothuria scabra
- Thi gian thu mu: thng 2 nm 2006
v thng 8 nm 2007
- a im thu mu: Ct B, Hi Phng
- Ngi nh loi: TS. Cng Thung v
cc cng s, Vin Ti nguyn v Mi
trng Bin, VAST

Phn lp v tinh ch cc hp cht:

Mu Holothuria scabra (10kg kh) c ra sch nhiu ln loi mui v c,
sau thi nh v chit siu m bng metanol. Qu trnh phn lp cc hp cht c
trnh by c th s di y, kt hp gia cc phng php sc k ct Silicagen pha
thng, pha o YMC v sc k lp mng TLC. T 200 gam dch c metanol, em chit
phn b bng hexan v nc vi t l 1/1 thu c 80 gam dch chit. Tng t ln lt
chit vi cloroform, butanol v nc theo t l 1/1 v th tch thu c 70 gam dch
chit cloroform v 30 gam dch chit butanol, phn cn nc cn li l 20 gam. Phn
dch chit CHCl3 c tm vo 90 gam Silicagel, sau a ln ct v chy sc k vi
hn hp dung mi n-gexan/axeton c gradient phn cc tng dn t 0% n 100%
axeton, thu c 6 phn on khc nhau. Phn on s 3 tip tc c chy sc k ct
Silicagel pha thng vi h dung mi n-hexan/axeton 4:1 thu c 2 cht sch HS-7 (15
mg) v HS-8 (12 mg) u c dng tinh th hnh kim, mu trng. Nhm 7 cng c
chy sc k ct pha thng vi h n-hexan/axeton 10:1 thu c cc tinh th hnh kim
mu trng HS-3 vi lng 10 mg. T nhm 5 sau khi tin hnh sc k ct pha thng
bng h dung mi CHCl3/MeOH 2,3:1 thu c lng ln hai cht bt mu trng, k
hiu l HS-1 v HS-2. Nhm 4 cng c tin hnh sc k nh nhm 5, thu c cht
HS-6 dng bt mu trng. Bng cc phng php tng t, cc hp cht HS-3 v HS9 c phn lp t cn butanol, HS-4 v HS-5 t cn nc (s IV.1).

54

Hi sm Holothuria
scabra (10 kg)

Thi nh, chit siu m trong

MeOH, ct loi dung mi
Dch chit MeOH
(200 g)
B sung nc
B sung CHCl3

Chit n-Hexan 1:1

Chit CHCl3 1:1

B sung n-BuOH

Chit BuOH 1:1

Cn n-Hexan
(80 g)

Cn CHCl3
(70 g)

Cn n-BuOH
(30 g)

CC, Sephadex
CHCl3/MeOH/H2O
1:4:1

CC, silicagel, n-hexan/axeton

gradien phn cc tng dn

10

11

12

HS-4 (9 mg)

HS-9 (12 mg)

HS-3 (10 mg)

CC, Silicagel, YMC, HS-5 (13 mg)

Axeton/H2O 1/1

CC, Silicagel, YMC,

MeOH/H2O 1/3

CC, Silicagel, YMC,

Axeton/H2O 1/1

HS-6 (9 mg)

CHCl /MeOH 2/1

HS-1
(250 mg)
HS2 (220 mg)
3/

CHCl3//MeOH 9/1

n-hexan/axeton 4/1

HS-7 (15 mg)

HS-8(12 mg)

S IV.1. S qu trnh phn lp cc hp cht t

hi sm Holothuria scabra

55

100% MeOH

CC, Dianion
MeOH/H2O
70% MeOH

20% MeOH

100% axeton

80% axeton

40% n-hexan

60% n-hexan

80% n-hexan

100% n-hexan

Cn pha nc
(20 g)

Xc nh cu trc ha hc v hot tnh sinh hc ca cc hp cht:

Cu trc ha hc ca cc hp cht c xc nh bng cch phn tch chi tit cc
d kin v hng s vt l, ph cng hng t ht nhn, ph khi lng v so snh cc s
liu vi cc ti liu c cng b. Do khun kh ca bo co tm tt c hn
chng ti ch chnh by y qu trnh xc nh cu trc ca mt hp cht in hnh
HS4:
Hp cht HS4 c phn lp t pha nc, c dng bt mu trng. Cng thc phn
t ca HS4 c xc nh l C54H85O28SNa thng qua kt qu ph khi lng ESI-MS ti
m/z 1259,3 [M + Na]+ (positive) v 1235,5 [M - H]+ v ph khi lng phn gii cao HR
FAB-MS: m/z 1259,4900 [M + Na]+ (positive) v kt qu tnh phn tch nguyn t cho
cng thc C54H85O28SNa2 l 1259,4743.

Hnh IV.1.a: Ph 1H-NMR ca HS4

O
O

HO
12

O O

17

OH

NaO3SO
CH3
O

CH2OH

CH2OH

OH

O O

OH
O

OCH3

OH

OH
OH

OH

Hnh IV.1.b: Cu trc ha hc ca HS4

56

OH
OH

Trn ph 1HNMR, vng trng mnh xut hin 7 tn hiu singlet ca 7 nhm
metyl bc 3 (H 0,81; 1,02; 1,05; 1,07; 1,08; 1,18; 1,56 ppm) v tn hiu doublet ca 1
nhm metyl bc 2 vi H 1,24 (3H, d, J = 6,0 Hz). Ngoi ra trong phn t cn tn ti
mt nhm metoxy cng hng ti H 3,49 (3H, s). Cng trn ph ny, vng trng yu
cn quan st c 1 proton olefin ca 1 ni i b th 3 ln (H 5,26 ppm) v cc tn
hiu doublet ca 4 proton anome ca 4 n v ng ti: H 4,32 (1H, d, J = 7,5 Hz),
4,34 (1H, d, J = 7,5 Hz), 4,38 (1H, d, 8,0 Hz), v 4,49 (1H, d, J = 7,5 Hz).

Hnh IV.1.c: Ph 13C-NMR ca HS4

Hnh IV.1.d: Ph 13C-NMR v cc ph DEPT ca HS4

57

Ph 13C-NMR v DEPT 135 ca HS4 cho thy s c mt ca 54 nguyn t

cacbon trong phn t, bao gm 9 nhm metyl, 11 nhm metylen, 24 nhm metin v 10
cacbon bc 4. Trong c 2 nhm cacbonyl vi tn hiu cng hng ti 172,6 ppm v
208,1ppm v 1 ni i b th 3 v tr [ 114,5(d)/152,8 (s)]. Bn nhm mtin vi tn hiu
cng hng ti 103,92, 103,86, 103,62, 102,95 c xc nh l cc cacbon anome
ca 4 gc ng. Ngoi ra cn c 24 cacbon khc cng lin kt trc tip vi xi, bao
gm: 17 nhm metin ( 87,5, 69,5, 81,4, 74,8, 74,2, 74,5, 74,2, 86,2, 70,1, 72,1, 87,9,
68,4, 76,1, 73,5, 85,9, 69,27, 76,8), 3 cacbon bc 4 ( 91,3, 86,2, 68,0 ppm), 3 nhm
metylen ( 63,0, 60,9, 60,7) v 1 nhm metoxy ( 59,8).

Hnh IV.1.e: Ph HSQC ca HS4

Gi tr chuyn dch ha hc ca cc cacbon c gn vi cc proton tng
ng thng qua ph HSQC. Kt qu chi tit c tng hp trong bng IV.1. Cng thc
phn t C54H85O28SNa ca HS4 cho php xc nh hp cht ny c khng no l 12,
trong khi cc ph NMR khng nh ch tn ti 3 lin kt (2 nhm cacbonyl v 1 ni
di kiu b th 3 v tr), iu ny chng t trong phn t c 9 cu trc vng, trong c
4 vng pyranosyl ca 4 gc ng.
Vi cc thng tin thu c, HS4 c nhn nh c cu trc kiu tritecpenglycosit, bao gm khung holasthane lin kt vi chui 4 phn t ng (xylozQuinovz- Glucoz-OMe Glucz). Kt qu ph 13C-NMR v DEPT phn chui 4
ng v s liu ph ti cc v tr t C-1 n C-21, t C-28 n C-30 ca khung
holasthane ca HS4 hon ton trng khp vi cc s liu cng b ca holothurin A2
(Ahmad, V.U. and Basha, A. (2000), Spectroscopic Data of Saponins - Triterpenoid Glycosides, CRC
Press, pp. 372-373.). S khc bit gia 2 hp cht ny nm phn nhnh bn t v tr C-22
n C-27, c th so vi holothurin A2 th HS4 c thm 1 nhm cacbonyl v 1 nhm
ancol bc 3.
58

Hnh IV.1.f. Ph HMBC ca HS4

21

O
24

18

O
22

HO

4'

OH

CH3

4''

6'''

6''''
CH2OH

O O

15

29

28

O O
1''

2''
OH

O
4''''

2'

OH

CH2OH

OH

6
1'

NaO3SO

17

30

O O

13

26

5'

OH
12

19

27
25

4'''

1'''

OH
OCH3

1''''

OH

OH
3''''

OH

Hnh IV.1.g. Mt s tng tc HMBC chnh trong phn t HS4

Da vo cc tng tc xa trn ph HMBC, cc v tr ca nhnh bn t C-22 n C27 c xc nh. Proton H-21 ngoi cc tng tc vi C-20 v C-17, cn tng tc
vi C-22 (C 208,1 ppm), iu ny khng nh v tr 22 l nhm cacbonyl. Bn cnh
l tng tc ca H-23 vi C-22; H-24 vi C-23, C-25, C-26 v C-27. C-25 l cacbon bc
4 lin kt vi 2 nhm CH3 (v tr 26 v 27) v 1 nhm OH. Ngoi vic tng tc qua li
vi nhau, proton ti cc v tr 26, 27 cn tng tc vi C-25 v C-24. c bit proton
ca OH-25 (H 4,23, s) cng th hin tng tc vi C-26; C-27; C-25 v C-24. Hn na
cc gi tr ph ca nhnh ny cng hon ton trng khp vi phn nhnh 22-one-25hydroxyl isohexan trong ti liu (Kanchanapoom T. et al., Phytochemistry, 2002, 59, 215-228)
59

Bng IV.1. Kt qu ph NMR ca HS4

C

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
OH-17
OH-25
Xyl 1
2
3
4
5
Qui 1
2
3
4
5
6
Glc 1
2
3

35,7
26,2
87,5
38,9
51,9
20,6
27,4
40,0
152,8
39,7
114,5
69,5
57,3
45,2
37,8
35,8
91,3
172,6
21,9
86,2
20,6
208,1
33,5
36,4
68,0
29,1
29,4
16,1
27,4
19,0

103,8
81,4
74,8
74,2
63,0
103,6
74,5
74,2
86,2
70,1
17,3
102,9
72,1
87,9

Ha, c

HMBC

1,41 - 1,74 (2H, m)

1,63 - 1,85 (2H, m)
3,34 (1H, m)

C-1 Xyl

0,88 (1H, m)
1,67 (2H, m)
1,35 - 1,67 (2H, m)
2,87 (1H, m)

C-10

5,26 (1H, d, 4,5 Hz)

4,48 (1H, m)

C-13; C-8

1,69 2,05 (2H, m)

1,70 (2H, m)

C-14

1,05 (3H, s)

C-5, C-9, C-10

1,56 (3H, s)

C-17, C-20, C-22

2,64 (2H, m)
1,54 - 1,61 (2H, m)

C-22
C-23, C-25, C-26, C-27

1,07 (3H, s)
1,07 (3H, s)
0,81 (3H, s)
1,02 (3H, s)
1,18 (3H, s)
7,03 (1H, s)
4,23 (1H, s)
4,32 (1H, d, 7,5 Hz)
3,34 (1H, m)
3,07 (1H, m)
3,94 (1H, m)
3,20 - 3,93 (2H, m)
4,49 (1H, d, 7,5 Hz)
3,51 (1H, m)
3,29 (1H, m)
3,00 (1H, m)
3,33 (1H, m)
1,25 (3H, s)
4,38 (1H, d, 8,0 Hz)
3,21 (1H, m)
3,03 (1H, m)

C-24, C-25, C-27

C-24, C-25, C-26
C-3, C-4, C-5, C-29
C-3, C-4, C-5, C-28
C-8, C-14, C-15, C-13
C-13, C-15, C-17
C-24, C-25, C-26, C-27
C-3
C-1 Xyl
C-1 Xyl

60

C-1 Xyl, C-4 Xyl

C-2 Xyl
C-3 Qui
C-2 Qui
C-3, C-5, C-6 Qui, C-1Glc
C-4 Qui, C-5 Qui
C-4 Qui

4
5
6
OMeGlc
1
2
3
4
5
6
OCH3
OH-3

68,4
76,1
60,7

3,21 (1H, m)
3,31 (1H, m)
3,41 - 3,68 (2H, m)

103,9
73,5
85,9
69,3
76,8
60,8
59,8

4,34 (1H, d, 7,5 Hz)

3,15 (1H, m)
2,99 (1H, m)
3,14 (1H, m)
3,21 (1H, m)
3,41 - 3,68 (2H, m)
3,49 (3H, s)
5,80 (1H, s)

C-4 Glc
C-3 Glc
C-1 OMeGlc
C-2 OMeGlc, OCH3
C-5 OMeGlc

C-3 OMeGlc
C-2 Xyl, C-3 Xyl

o trong DMSO; bti 125MHz; cti 500MHz

Trn khung holosthan, cc tng tc gia H-5 vi C-10; gia H-11 vi C-13; C-8;
gia H-15 vi C-14; gia H-19 vi C-5, C-9, C-10; gia H-28 vi C-3, C-4, C-5, C-29;
gia H-29 vi C-3, C-4, C-5, C-28; gia H-30 vi C-8, C-14, C-15, C-13 cng c ghi
nhn. ng ch l OH-17 (H 7,03, s) cng th hin tng tc vi C-13, C-15 v C-17.

Hnh IV.1.h. Ph khi lng ESI-MS ca HS4

chui ng, th t lin kt MeGlc(1-3)-Glc(1-4)-Qui(1-2)-Xyl(1-3)-Aglycon
cng c xc nhn. Trc tin l tng tc xa H-3 vi C1xyl; H-1xyl vi C-3, khng
nh khung holosthan gn trc tip vi ng xylose. Tip theo l tng tc gia prton
anome H-1 Quino vi C-2Xylo; H-1Gluco vi C-4Quino; gia H-4Quino vi C1Gluco v H-1OMeGluco vi C-3Gluco. Trt t cc v tr trong tng gc ng
cng c lm sng t nh cc tng tc xa. gc OMeGluco xut hin cc tng tc
ca: H-OMe vi C-3; H-4 vi C-5; H-3 vi C-OMe v C-2, H-2 vi C1, cn gc Gluco l tng tc: H-6 vi C-4. ng Quino c tng tc H-6
vi C-5 v C-4; H-4 vi C-3, C-5, C-6; H-3 vi C-2; H-2 vi C-3 v H-1
vi C-2. Cui cng l ng Xylo c cc tng tc H-5 vi C-1, C-4; H-3 vi C-1
61

v H-2 vi C-1. V tr nhm OH-3 ca ng xyloz cng c xc nhn bng cc

tng tc OH-3 (5.80 ppm) vi C-2 v C-3.

Hnh IV.1.i. Ph khi lng phn gii cao HR-ESI-MS ca HS4

T tt c cc phn tch nu trn, hp cht HS4 c xc nh l 3,12,17,25tetrahydroxyholost-9(11)-ene-22-one 3-O-[(3-O-methyl)--D-glucopyralnosyl-(13)-D-glucopyrano-syl-(14)--D-quino-vopyranosyl-(12)-(4-O-sunfo)--Dxylopyranoside]natri. y l mt hp cht mi, ln u tin phn lp c t thin
nhin v c chng ti t tn l holothurin A3
Bng cc phng php tng t, chng ti xc nh c cu trc ha hc ca
ton b cc cht c phn lp t loi hi sm Holothuria scabra. Cc hp ny c
tin hnh nh gi hot tnh sinh hc theo cc phng php c trnh by phn
phng php. Kt qu chi tit c tm tt sau y:
1. Hp cht HS1: holothurin B
- Cu trc ha hc:
21

O
HO

19

10

Xyl
4'

NaO3SO
6'' CH3

Qui
4''

OH

17
13

26

25
27

OH

15
30

O O

OH

22
20

12

11

3
5'

O
18

24

23

6
1'

28

29

2'
O O
1''

OH
2''

OH

- Cht bt v nh hnh mu trng: mp 251-253oC

- Cng thc phn t: C41H63O17NaS (M = 882)
- Ph khi lng: m/z 905 [M+Na]+
62

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng KB: IC50 = 2,85 g/ml
Dng FL: IC50 = 2,14 g/ml
Dng Hep-G2: IC50 = 1,99
g/ml
+ Hot tnh khng VSV kim
nh
Khng cc chng nm:
Aspergillus niger, Fusarium
oxysporum, Candida
albicans, v Saccharomyces
cerevisiae vi gi tr MIC =
50 g/ml

2. Hp cht HS2: holothurin A

- Cu trc ha hc:
O

HO
19

17

12

10

Xyl
4'

15

2'

CH2OH

Glc

4'''

6''''

OH

MeGlc
4''''

OH

2''

OH
1'''
2'''

OH

1''''

OCH3

O O

CH2OH

- Ph khi lng: m/z 1243,3 [M+Na]+

- Cht bt v nh hnh mu trng: mp 228-230oC
- Cng thc phn t: C54H85O27SNa (M=1220)
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 4,08 g/ml
Dng KB: IC50 = 1,28 g/ml
Dng FL: IC50 = 1,54 g/ml

1''

OH

6'''

6
29

28

O O

Qui
4''

27

1'

NaO3SO
6'' CH3

25

30

O O

OH

26

OH

13

3
5'

22
20

18

11

24

23

21

2''''

OH

3. Hp cht HS3: holothurin A2

- Cu trc ha hc:
21

O
HO
19

10

4'

6'' CH3

4''
6'''

CH2OH

CH2OH
O

O
4''''

OH

1''''

OCH3

2''''

OH

2''

1'''

OH

1''

OH

4'''

2'''

OH

23

OH
27

15

6
28

29

2'
O O

OH

O O

6''''

26
25

1'

NaO3SO

24

30

O O

OH

17
13

3
5'

20

18
12

11

22

- Cht bt v nh hnh mu trng: mp 246-2480 C

- Cng thc phn t: C54H87O26SNa, M = 1206
- Ph khi lng: m/z 1229,6 [M+Na]+
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng KB: IC50 = 2,42 g/ml
Dng FL: IC50 = 1,00 g/ml
Dng Hep-G2: IC50 = 1,06 g/ml

63

4. Hp cht HS4: holothurin A3 (Cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
21

Xyl
4'

10

Qui

4''

CH2OH

CH2OH

MeGlc
4''''

OH

OH

15

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng FL: IC50 = 0,39 g/ml
Dng Hep-G2: IC50 = 0,23 g/ml
Dng KB: IC50 = 0,57 g/ml

1'''

OH

1''''

OCH3

4'''

27

(tnh ton l thuyt cho C54H86O28NaS: 1237,4924)

2''

Glc

OH

- Cht bt v nh hnh mu trng

- Cng thc phn t: C54H85O28NaS, M = 1236
- Ph khi lng: m/z 1259,5 [M+Na]+, 1235,5 [M-H]- Ph khi lng phn gii cao: m/z 1237,5027 [M+H]

1''

O O

6''''

OH

29

28

O O

OH

6'''

25

2'

6'' CH3

23

1'

NaO3SO

26

30

O O

OH

17
13

3
5'

20

18
12

11

24

22

HO
19

2'''

OH

2''''

OH

5. Hp cht HS5: holothurin A4 (Cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
OH

21

O
HO
19

10

4'

1'

NaO3SO
6'' CH3

4''
6''''

6'''

CH2OH

O O

O
1''''
4''''

OCH3
OH

2''''

OH

1'''

OH

1''
2''

OH

4'''

2'''

OH

26
25

23

OH
27

15

4
6
28

29

2'
O O

OH

CH2OH

17

24

30

O O

OH

20

18

13

3
5'

12

11

22

- Cht bt v nh hnh mu trng

- Cng thc phn t: C54H87O27SNa (M=1222)
- Ph khi lng: m/z 1245,2 [M+Na]+, 1221,4 [M-H]- c phn lp t loi: Holothuria scabra.
- Hot tnh sinh hc:
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 0,57 g/ml
Dng KB: IC50 = 1,12 g/ml

64

6. Hp cht HS6: holothurigenin B (Cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
23

24

21

25

17

12

11

10

4'

OH

29

28

O
1''

OH

OH

30

2'
O

4''

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 2,33g/ml
+ Hot tnh khng VSV kim nh
Khng cc chng nm: Aspergillus
niger, Fusarium oxysporum,
Candida albicans, v
Saccharomyces cerevisiae vi gi
tr MIC = 50 g/ml

15

6
1'

- Cng thc phn t: C41H62O13 (M =

762)
- Ph khi lng: m/z 762,7 [M+H]+,
785,3 [M+Na]+, 761,3 [M-H]-

O O

OH
6'' CH3

27

OH

13

3
5'

20

18
19

26
22

2''

OH

- Cht bt v nh hnh mu trng

7. Hp cht HS7: 2,3,12,21,24R,25-hexahydroxylanosta-8-en

- Cu trc ha hc:
OH

21

OH

18

2
10

HO

30

H
28

15

OH
27

- Tinh th hnh kim mu trng

- Cng thc phn t: C30H52O6 (M 508
- Ph khi lng: m/z 490 [M-H2O]+

16

14

26
25

17
13

19
1

HO

OH
24

23

20

12
11

22

29

8. Hp cht HS8: 24R-ergosta-7,22-dien-3,5,6-triol

- Cu trc ha hc:
28
21

22

18

24

20

12
13

19
1

14

10

HO

OH

15

- Ph khi lng: m/z 412 [M-H2O]+

16

- Tinh th hnh kim mu trng

- Cng thc phn t: C28H46O3, M =
430.

27

17

11

26
25

23

7
6

OH

9. Hp cht HS9: Hp cht -Adenosine

- Cu trc ha hc:
NH 2
N

- Tinh th hnh kim mu trng:

mp 234-235oC
- Cng thc phn t: C10H13N5O4, M
= 267
- Ph khi lng: m/z 267,8 [M +
H]+, 289,8 [M + Na]+

HO
O
(R)

OH

OH

(R)

65

Kt qu th nghim hot tnh cho thy: Tt c cc hp cht hothurin u th hin

hot tnh gy c t bo v cc hp cht cn li khng c biu hin hot tnh. iu ny
th hin s ph hp vi cc cng trnh nghin cu c cng b trn th gii bi lp
cht ny c hot tnh gy c t bo c trng. Cc hp cht HS1, HS2, HS3 v HS4
th hin hot tnh trn c 3 dng t bo th nghim, hp cht HS5 th hin hot tnh trn
hai dng t bo th nghim l Hep-G2 v KB, v hp cht HS6 ch th hin hot tnh
trn dng t bo Hep-G2.
Ngoi ra, 2 hp cht HS-3 v HS-4 c th nghim hot tnh theo phng php
quan st trc tip di knh hin vi. Tc dng gy c t bo ca cc hp cht HS-3 v
HS-4 c quan st v chp nh trn knh hin vi sau cc thi im 24h, 48h, 72h. Do
khun kh bo co c hn, di y chng ti ch a ra mt s hnh nh i din:
Sau 24 gi

Mu i chng m

Nng 1 g/ml

Nng 5 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml
Hnh nh t bo vi mu th l HS-3 cc nng khc nhau
66

Nng 5 g/ml

Nng 1 g/ml

Nng 0,2 g/ml

Nng 0,04 g/ml

Hnh nh t bo vi mu th l HS-4 cc nng khc nhau

T cc hnh nh t bo chp c qua knh hin vi chng ti c nhn xt nh sau:
- C 2 mu HS-3 v HS-4 u c kh nng c ch s phn chia v tng trng
ca t bo, c ch s to si (antimitotic activity) v to mng lin kt gia cc t bo
(anti-angiogenic activity) thng qua s thay i r rt v hnh thi t bo, b mt mng
t bo v kh nng to mng si bm dnh vo y phin th nghim.
- Mu HS-3 c tc dng r rt hn biu hin s thay i v hnh thi mng t
bo v hnh nh pht trin ca t bo: sau 24 gi th nghim th nng cht 5g/ml
t bo gn nh khng pht trin v s lng, t bo bm dnh l rt t. Sau 48 v 72 gi
lng t bo bm dnh khng cn, t bo b bong ra khi y phin th nghim v
cht.
- Cc kt qu v hnh thi cng lm sng t hn cho kt qu th nghim gy c
t bo trn phin vi lng 96 ging sau 72 gi. Vi th nghim gy c t bo, cc mu
HS-3 v HS-4 u cho kt qu dng tnh mnh trn c 3 dng t bo th.
Cc kt qu thu c (Th nghim gy c t bo v quan st hnh thi b mt
mng t bo) gi cho cc nghin cu tip theo mc cao hn mc phn t mng
t bo v tc dng gy ging ho- cht theo chng trnh (apoptosis) c th lm r
hn v c ch tc ng ca cc cht trn.
Kt qu nh gi hot tnh khng vi sinh vt kim nh cho thy: Ch c hp cht
HS1 v HS6 th hin hot tnh khng vi sinh vt kim nh. Mt iu th v l cc hp
67

cht ny khng rt mnh tt c cc chng nm vi gi tr MIC u t mc thp l 50

g/ml, trong khi chng li khng c biu hin hot tnh i vi cc loi vi sinh vt
khc. iu ny th hin s ph hp hon ton vi cc nghin cu cng b trn th
gii (hin ch c 1 ti liu cng b v hot tnh khng nm ca cc hp cht holothurin
(Kumar R, Chaturvedi AK, Shukla PK, Lakshmi V. Bioorg Med Chem Lett. 2007, 17(15), 4387-4391).
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc loi hi sm Holothuria
scabra, chng ti bin son v ng ti 2 cng trnh trn cc tp ch chuyn ngnh quc t v 01 cng
trnh trong hi ngh:
1. Nguyen Hai Dang, Nguyen Van Thanh, Phan Van Kiem, Le Mai Huong, Chau Van Minh and Young
Ho Kim, Two new triterpene glycosides from Vietnamese sea cucumber. Archives of Phamarcal
Research (Korea), 2007, 30(11), 1387-1391.
2. Nguyen Van Thanh, Nguyen Hai Dang, Phan Van Kiem, Nguyen Xuan Cuong, Hoang Thanh Huong,
and Chau Van Minh, A new triterpene glycoside form the sea cucumber Holothuria scabra collected
in Vietnam. Asean Journal on Science and Technology for Developtment, 2006, 23(4), 253-259.
3. Nguyen Van Thanh, Chau Van Minh, Hoang Thanh Huong, Phan Van Kiem. Steroidal constituents
from Holothuria scabra living in Vietnamese sea. Tuyn tp cc bi bo khoa hc Hi ngh Khoa hc
ln th 20-HBK H Ni, phn ban Cng ngh Ha hu c, 2006, 200-204.
Ngoi ra cng t cc kt qu ny, tp th tc gi xy dng v ng k 01 bng pht minh sng
ch ti Cc S hu Tr tu Vit Nam l: Chu Vn Minh, Phan Vn Kim, Nguyn Hi ng, Nguyn
Vn Thanh, L Mai Hng, Cng Thung, Trn Thu Hng, Nguyn Xun Cng. Hp cht
Holothurin A3 c hot tnh chng ung th v phng php chit hp cht ny t loi hi sm Holothuria
scabra, Bng c quyn sng ch s 6852, cp theo quyt nh s 2394/Q-SHTT, ngy 14 thng 02 nm
2008 ca Cc S hu Tr Tu Vit Nam
Cc kt qu nghin cu c tham gia gii thng khoa hc k thut thanh nin ln th XVIII 2008 v ot gii khoa hc: s 1890/GTKHKTTN-2008 theo quyt nh s 1890/Q-KHCNVN ca Ch
tch Vin Khoa hc v Cng ngh Vit Nam ngy 18/11/2008 vi ti: Nghin cu thnh phn ha hc,
hot tnh sinh hc v ng dng ca loi hi sm trng Holothuria scabra sinh sng ti cc vng bin Vit Nam.
Cc kt qu trn c s dng trong 01 Lun n Tin s bo v thnh cng (TS. Nguyn Vn
Thanh)

IV.2. nghin cu ha hc theo nh hng hot tnh sinh hc

loi sao bin Archaster typicus (archasteridae)

- Tn Vit Nam: Sao bin

- Tn khoa hc: Archaster
typicus
- Thi gian thu mu: thng 1
nm 2006 v thng 8 nm 2007
- a im thu mu: Ct B,
Hi Phng
- Ngi nh loi: TS. Cng
Thung v cc cng s, Vin Ti
nguyn v Mi trng Bin,
VAST

Phn lp cc hp cht:
Sao bin Archaster typicus (5 kg kh) c ra sch, xay nh v chit bng
metanol (3 ln) bng thit b chit siu m. Dch metanol ca cc ln chit c lc,
gom li v ct loi dung mi nhit khng qu 60oC di p sut gim thu c 200
g dch cn dch metanol. Cn dch ny c ha vo nc v chit phn b ln lt vi
cc dung mi clorofooc v butanol theo t l 1/1 (3 ln, mi ln 6 lit) thu c cc cn
dch tng ng l clorofooc 65 g, butanol 35 g v nc 100 g. Cn chit clorofrom (65
68

g) c tin hnh chy ct th trn sc k ct pha thng gradient nng axeton trong
hexan thu c 4 phn on nh hn l nhm 1 n 4. Cc phn on nhm 1, 2 v 3
c tip tc phn tch trn sc k ct pha thng vi h dung mi ra gii thch hp
thu c cc hp cht SB5, SB6 v SB7. Bng cc phng php tng t cc hp cht
SB1, SB2, SB3, v SB4 c phn lp t cn chit BuOH (S IV.2).
Sao bin Archaster
typicus (5 kg)
Thi nh, chit siu m trong
MeOH, ct loi dung mi
Dch chit MeOH
(200 g)
B sung nc
Chit CHCl3/H2O 1:1

B sung BuOH
Chit BuOH/H2O 1:1

Cn CHCl3
(65 g)

Cn BuOH
(35 g)

CC, silicagel, n-hexan/axeton

gradien phn cc tng dn

Nhm 6

Nhm 7

CHCl3//MeOH/H2O 6/1/0,1

SB-4 (18 mg)

SB-3 (9 mg)
SB-2 (8 mg)

SB-1 (18 mg)

SB-7 (14 mg)

SB-6 (25 mg)

69

Nhm 8

CC, Silicagel, YMC,

Axeton/MeOH/H2O 1/8/1

CHCl3//MeOH/H2O 8/1/0,1

n-hexan/axeton 5/1

Kt tinh trong
axeton

n-hexan/axeton 15/1

SB-5 (13 mg)

S IV.2. S phn lp cc hp cht t sao bin

100%

Nhm 5

35% CHCl3

Nhm 4

70% CHCl3

Nhm 3

100% CHCl3

Nhm 2

CC, silicagel, CHCl3/MeOH gradien

phn cc tng dn
100% axeton

35% n-hexan

70% n-hexan

100% n-hexan

Nhm 1

Cn pha nc
(100 g)

Xc nh cu trc ha hc v hot tnh sinh hc cc hp cht:

Tng hp cu trc ha hc v HTSH ca cc cht c phn lp t loi sao bin
Archaster typicus c a ra di y:
1. Hp cht SB1: archastercerebroside (cht mi, ln u tin c phn lp t thin
nhin)
- Cu trc ha hc:
- Ph khi lng phn gii cao:
OH
OH OH
6"

4"

H
HO

H O

5"

2"

3"

(CH2)21CH3

NH

1"

OH
H

2'

1'

m/z 840,6734 [M+Na]+ (tnh

ton l thuyt cho
C46H91NO10Na: 840,6541)

OH
4

(CH2)9CH(CH3)2

Gal

OH

- Cht bt v nh hnh mu trng: mp 221-222oC

- Cng thc phn t: C46H91NO10 (M = 817)
- Ph khi lng: m/z 818,8 [M+H]+, 816,8 [M-H]-

- Hot tnh sinh hc:

+ hot tnh khng NF-B
IC50 = 34,7 1,84 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 4,45 g/ml
Dng FL: IC50 = 3,91 g/ml
Dng Hep-G2: IC50 = 2,35
g/ml

2. Hp cht SB2: 27-norcholestan-3,4,5,6,8,14,15,24R-octol

- Cu trc ha hc:
OH

21

22

12
11

19
1

10

OH

15

OH

HO

16
14

OH

- Hot tnh sinh hc:

+ Hot tnh khng NF-B:
IC50 = 30,82,12 M
+ Hot tnh gy c t bo
Dng KB: IC50 = 3,37 g/ml
Dng Hep-G2: IC50 = 2,69
g/ml

26

25

23

17

13

OH
8

24

20

18

OH

OH

- Tinh th hnh kim mu trng: mp 294oC

- Cng thc phn t: C26H46O8 (M = 486)
- Ph khi lng: m/z 509,2 [M+Na]+, 485,4 [M-H]3. Hp cht SB3: 27-norcholestan-3,4,5,6,7,8,14,15,24R-nonol
- Cu trc ha hc:
OH

21

22
20

18
12
11

19
1

HO

10

26

23

16
14

OH

15

OH

OH

25

17

13

OH

24

OH

OH
OH

- Tinh th hnh kim mu trng: mp 288-290oC

- Cng thc phn t: C26H46O9 (M = 502)
- Ph khi lng: m/z 501,3 [M-H]-, 525,2 [M+Na]+
70

- Hot tnh sinh hc:

+ Hot tnh khng NF-B:
IC50 = 28,11,98 M
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 4,52
g/ml

4. Hp cht SB4: ergost-22-en-3,4,5,6,8,14,15,22E,24R,25R,26-nonol

- Cu trc ha hc:
28
- Cng thc phn t: C28H48O9 (M =
21

22

20

18
12
11

19
1
2

9
10

HO

OH

OH

OH

528)

- Ph khi lng: m/z 527,4 [M-H]+,

551,3 [M+Na]+

- Hot tnh sinh hc:

+ Hot tnh khng NF-B:
IC50 = 27,81,87 M
+ Hot tnh gy c t bo
Dng Hep-G2: IC50 = 3,72 g/ml

15

OH

OH
27

16
14

23

17

13

OH

26

CH 2 OH

25

24

OH

- Tinh th hnh kim mu trng: mp 288-290oC

5. Hp cht SB5: cholesterol
- Cu trc ha hc:
21

21

20

24

25

- Tinh th hnh kim mu trng:

mp 148-149oC
- Cng thc phn t: C27H46O,
M=386

18
17

26

19
1
8
3
5

HO

6. Hp cht SB6: cholest-7-en-3-on

- Cu trc ha hc:
21

22

18

19

12

11

13
14

2
3

1
10
5

20

24

26
25

23

27

17
16
15

8
7

- Tinh th hnh kim mu trng:

mp 147-148oC
- Cng thc phn t: C27H44O, M =
384
- Ph khi lng: m/z 385,2
[M+H]+

7. Hp cht SB7: batyl alcohol (batilol)

- Cu trc ha hc:
1

- Tinh th mu trng: mp 70-71oC

- Cng thc phn t: C21H44O3, M =
344
- Ph khi lng: m/z 345,1
[M+H]+

OH

HO

1'

18'

CH2(CH2)16CH3

Cc hp cht phn lp c t loi sao bin Archaster typicus c tin hnh th

nghim hot tnh khng NF-B, mt yu t phin m c tm quan trng c bit trong
cc p ng min dch v vim. NF-B iu tit s biu hin ca nhiu gen tham gia vo
cc p ng min dch v vim. N lm tng s biu hin ca cc gen quy nh nhiu
cytokine, cc enzym v cc phn t dnh kt trong cc bnh vim mn tnh. NF-B c
hot ha bi nhiu yu t khc nhau (virus, cc cht xy ha v cc khng nguyn) lm
71

tng p ng vim. S hot ha ny li dn ti s biu hin phi hp ca nhiu gen m

ha cc protein (nh cc cytokine, chemokine, enzym v cc phn t dnh kt) tham gia
vo s tng hp cht trung gian v ko di thm p ng vim. V vy, NF-B l mt
ch hin nhin cho cc kiu iu tr khng vim mi. Glucococticoit l cc cht c ch
NF-B hu hiu nhng chng c nhiu tc dng ph ti ni tit v chuyn ha khi c
s dng vi liu php iu tr ton thn. Cc tc dng ph ny s khng xy ra khi s
dng cc cht c ch NF-B c hiu hn. V vy, vic tm kim v nghin cu k lng
cc hp cht thin nhin c tc dng c ch c hiu NF-B v ang thu ht mnh
m s quan tm v u t nghin cu ca cc nh khoa hc trn ton th gii.
Kt qu nghin cu ca chng ti cho thy, 04 hp cht SB1 - SB4 th hin hot
tnh khng NF-B tt. y u l cc hp cht c cu trc c o, c trng cho loi
sao bin Archaster typicus (c th s dng chng nh yu t ch th trong phng php
nh loi ha hc). Cc hp cht ny c phn lp vi hm lng cao trong loi sao
bin Archaster typicus, iu ny t c s cho vic tin hnh cc nghin cu ton din
hn loi sao bin ny theo nh hng hot tnh khng NF-B. Mt iu rt ng quan
tm na l SB1 l mt hp cht c cu trc mi v c phn lp vi hm lng cao
nht so vi cc cht cn li, iu ny gi m hng nghin cu, pht trin cc dc cht
mi khng yu t phin m NF-B v c s dng iu tr cc bnh vim, gp phn
nng cao gi tr s dng ca ngun nguyn liu sinh vt bin qu bu v sn c ca
nc ta.
Ngoi ra, tt c cc cht phn lp c t loi sao bin Archaster typicus cn c
tin hnh nh gi hot tnh gy c t bo. Kt qu th nghim cho thy c 4 hp cht
SB1 - SB4 th hin hot tnh gy c t bo. Trong hp cht SB1 th hin hot tnh
gy c t bo i trn c 3 dng t bo th nghim, SB2 khng 2 dng t bo KB v
Hep-G2 vi gi tr IC50 ln lt l 3,37 v 2,69 g/ml, hai hp cht cn li SB3 v SB4
ch khng dng Hep-G2 vi gi tr IC50 ln lt l 4,52 v 3,72 g/ml
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc loi sao bin Archaster
typicus, chng ti bin son v ng ti 02 cng trnh trn cc tp ch chuyn ngnh quc gia, 01 cng
trnh ti Hi ngh khoa hc quc gia v 01 cng trnh ti Hi ngh khoa hc quc t l:
(1) Chau Van Minh, Nguyen Xuan Cuong, Nguyen Van Thanh, Hoang Thanh Huong, and Phan Van
Kiem, Highly oxygenated sterols from the starfish Archaster typicus. Tp ch Ha hc, 2007, 45(3),
377-381.
(2) Phan Van Kiem, Nguyen Van Thanh, Nguyen Xuan Cuong, and Chau Van Minh, Cholest-7-en-3-one,
cholesterol and batilol from the starfish Archaster typicus. Tp ch Ha hc, 2007, 45(4), 513-517.
(3) Chu Vn Minh, Nguyn Vn Thanh, Nguyn Xun Cng, Phan Vn Kim, Nunziatina De Tommasi,
Lm Ngc Thim. Mt galactocereboside mi t sao bin Archaster typicus. Hi ngh Ha hc hu c
Ton quc ln th 4, H Ni, thng 10 nm 2007, trang 446-450.
(4) Chau Van Minh, Nguyen Van Thanh, Nguyen Xuan Cuong, Phan Van Kiem, Nunziatina De Tommasi.
A new galactocereboside from the starfish Archaster typicus, 12t Asian Chemical Congress (12ACC),
August 23-25, 2007, Malaysia, p. 309.
Cc kt qu trn c s dng trong 01 Lun n Tin s bo v thnh cng vi ti:
Nghin cu thnh phn ho hc v kho st hot tnh gy c t bo in vitro ca loi hi sm Holothuria
scabra v sao bin Archaster typicus sinh sng ti vng bin vit nam (TS. Nguyn Vn Thanh)

72

Chng V. Nghin cu ho hc theo nh hng hot tnh

sinh hc ca mt s loi Hi min
V.1. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi
hi min xestospongia testudinaria (petrosiidae)

- Tn Vit Nam: Hi min

xp
Tn
khoa
hc:
Xestospongia testudinaria
- Thi gian thu mu: thng
3 nm 2007
- a im thu mu: Nam
Yt, Qun o Trng Sa,
tnh Khnh Ha
- Ngi nh loi: TS.
Cng Thung v cc cng
s, Vin Ti nguyn v
Mi trng Bin, VAST

Phn lp v tinh ch cc hp cht:

Hi min Xestospongia
testudinaria ti (15 kg)
- Chit MeOH, ct loi dung mi di p sut
gim
B sung nc
B sung clorofooc

GM - Cn chit MeOH (120g)

clorofooc: nc 1/1

Lp nc
B sung BuOH

Cn clorofooc
C (60 g)

BuOH: nc 1/1

Lp nc

Cn nc
N (20 g)

Cn BuOH
B (40 g)

S V.1.a: S chit phn on mu hi min

73

Hi min X. testudinaria (15 kg ti) c ra sch, xay nh v chit bng

metanol (3 ln) bng thit b chit siu m. Dch metanol ca cc ln chit c lc,
gom li v ct loi dung mi nhit khng qu 60oC di p sut gim thu c 120
g dch cn dch metanol. Cn dch ny c ha vo nc v chit phn b ln lt vi
cc dung mi clorofooc v butanol theo t l 1/1 (3 ln, mi ln 6 lit) thu c cc cn
dch tng ng l clorofooc 60 g (k hiu: C), butanol 40 g (k hiu B) v nc 20 g (k
hiu N) (s V.1.a).
C (60 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

C1 (30g)

C2 (5 g)

C3 (7 g)

C4 (3 g)

C5 (15 g)

Silica gel CC
n-hexan/ axeton 4/1

XT9
(12mg)

XT10
(8mg)

Silica gel CC
n-hexan/clorofooc/axeton 6/1/0,3

XT4
(20 mg)

XT5
(80 mg)

XT6
(35 mg)

XT2
(30 mg)

XT3
(1200 mg)

Silica gel CC
metanol/nc 5/1

XT1
(20 mg)

S V.1.b: S phn lp cc hp cht t cn chit cloroform

Cn chit clorofrom: C (60 g) c tin hnh chy ct th trn sc k ct pha
thng gradient nng metanol trong cloroform thu c 5 phn on nh hn l C15. Cc phn on C1, C2 v C3 c tip tc phn tch trn sc k ct pha thng vi h
dung mi ra gii thch hp thu c cc hp cht XT1 n XT6 v XT9 - XT10. Bng
cc phng php tng t, cc hp cht XT7, v XT8 c phn lp t cn chit BuOH
(S V.1.b-c).

74

B - (40 g)
Silica gel CC
Cloroform/MeOH/H2O: 5/1/0 - 2/1/0.2

B1 (1.0 g)

B2 (20 g)

Silica gel CC
CHCl3/MeOH/H2O
5/1/0.1

B3 (3 g)

B4 (6 g)

B5 (10 g)

Silica gel CC
CHCl3/MeOH/H2O
4/1/0.1

XT8
(33 mg)

XT7
(40mg)

S V.1.c. S phn lp cc hp cht t cn chit butanol

Xc nh cu trc ha hc v hot tnh sinh hc cc hp cht:
Cu trc ha hc ca cc hp cht c xc nh bng cch phn tch chi tit cc
d kin v hng s vt l, ph cng hng t ht nhn, ph khi lng v so snh cc s
liu vi cc ti liu c cng b. Do khun kh ca bo co tm tt c hn
chng ti ch chnh by y qu trnh xc nh cu trc ca mt hp cht in hnh
XT1:
Hp cht XT1 phn lp c di dng tinh th hnh kim mu trng. Cng thc
phn t ca n c xc nh l C20H21BrO2 (M = 372) bng cc kt qu ph khi lng
(ESI-MS ti m/z 394,8/396,8 [M+Na]+ vi t l 1/0,98 v cng tn hiu) v cc ph
cng hng t ht nhn (1H-, 13C-NMR v cc ph DEPT). Hp cht ny rt km bn
nn i hi phi c o ph khi lng v ph cng hng t ht nhn ngay sau khi
c lm sch.
Trn ph 1H-NMR xut hin cc tn hiu c trng ca ba ni i ngoi vng u
b th hai v tr ti 5,52 (1H, d, J = 15,5 Hz, H-11), 5.57 (1H, dd, J = 15.5, 1.5 Hz, H16), 6.13 (1H, m, H-15), 6.23 (1H, m, H-12), 6.29 (1H, dd, J = 14.0, 2.5 Hz, H-19) v
6.65 (1H, d, J = 14.0 Hz, H-20). S xut hin ca by nhm metilen c c trng bi
cc tn hiu cng hng ti 1,47 (2H, m, H-4), 1,57 (2H, m, H-5), 1,65 (2H, m, H-3),
2,23 (4H, m, H-13 v H-14), 2,33 (2H, t, J = 7,0 Hz, H-6) v 2,36 (2H, t, J = 7,5 Hz, H2).
6
7
9
17
15

Br

11

19

Hnh V.1.a. Cu trc ha hc ca XT1

75

COOH
1

Hnh V.1.b. Ph 1H-NMR ca XT1

Ph 13C-NMR ca XT1 xut hin cc tn hiu ca 20 cacbon trong c 7 cacbon
bc bn, 7 cacbon metilen v 6 cacbon metin. Ba lin kt i ngoi vng mt ln na
c khng nh bi cc tn hiu cacbon metin ti 109.84 (C-11), 110,32 (C-16),
117,64 (C-19), 117,89 (C-20), 143,83 (C-15) v 146,02 (C-12). Ngoi ra, s xut hin
ca 3 lin kt ba c c trng bi cc tn hiu u l cacbon bc bn cng hng ti
65,40 (C-9), 73,43 (C-10), 73,68 (C-8), 83,52 (C-7), 85,04 (C-18) v 90,22 (C-17). Mt
nhm cacbonyl c xc nh bi tn hiu cng hng ti 179,29 (C-1). Cc nhm
metilen nm v tr v so vi nhm cacbonyl c khng nh bi cc tn hiu cng
hng tng ng ti 33,75 (C-2) v 24,12 (C-3). Mt nhm metilen nm cnh ni ba
v 2 nhm metilen nm cnh ni i c khng nh bi cc tn hiu tng ng ti
19,38 (C-6), 32,15 (C-13 v C-14).

Hnh V.1.c. Ph 13C-NMR ca XT1

76

Hnh V.1.d. Ph 13C-NMR v cc ph DEPT ca XT1

T cc phn tch nu trn, c th s b xc nh XT1 l mt hp cht axit mch
di b brm ha c cha 3 ni i v 3 ni ba. Cu trc hnh hc ca cc ni i c
xc nh u dng trans bi hng s tng tc ln (J = 14,0 v 15,5 Hz) ca cc
proton.

Hnh V.1.e. Ph HSQC ca XT1

77

Bng V.1. Kt qu ph NMR ca XT1

C

C#

Ca, b

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20

179,4
36,0
25,7
19,7
83,7
74,4
66,5
74,0
111,2
147,4
33,1
33,1
145,3
110,0
91,1
85,6
118,7
118,9

179,29
33,75
24,12
28,19
27,87
19,38
83,52
73,68
65,40
73,43
109,84
146,02
32,15
32,15
143,83
110,32
90,22
85,04
117,64
117,89

Ha, c
multiplicity (J in Hz)
2,36 t (7,5)
1,65 m
1,47 m
1,57 m
2,33 t (7,0)
5,52 d (15,5)
6,23 m
2,23 m
2,23 m
6,13 m
5,57 dd (15,5, 1,5)
6,29 dd (14,0, 2,5)
6,65 d (14,0)

HMBC
(H C)
1, 3, 4
1, 2, 3
2, 5, 6
3, 4, 6, 7
5, 7, 8

9, 10, 12, 13
10, 13
11, 12, 14
13, 15, 16
14, 17
14, 15, 18
17, 20
18, 19

o trong CDCl3, b125 MHz, c500 MHz, #s liu ca 18-bromo-(9E,13E,17E)-eicosa-9,13,17-triene5,7,15-triynoic acid.

Hnh V.1.f. Ph 1H-1H COSY ca XT1

V tr th ca nguyn t brm ti C-20 v s xp xp cc lin kt i v lin kt
ba ca XT1 nh c ch ra Hnh V.1.b c khng nh bng cch so snh s liu
78

ph NMR ca n vi cc s liu ph c cng b ca 18-bromo-(9E,13E,17E)eicosa-9,13,17-triene-5,7,15-triynoic acid (Ashok D. P., et al., Journal of Natural Products,
1992, 55(9), 1170-1177). iu ny mt ln na c khng nh bi cc tng tc trn ph
cng hng t ht nhn hai chiu HMBC v 1H-1H COSY. Cc phn tch chi tit c
ch ra trong Bng V.1 v Hnh V.1.h.

Hnh V.1.g. Ph HMBC ca XT1

6

COOH

9
17
15

Br

11

19

COSY
HMBC

Hnh V.1.h. Cc tng tc HMBC (HC) v COSY chnh ca XT1

Hnh V.1.i. Ph khi lng ESI-MS ca XT1

79

T tt c cc phn tch nu trn, cu trc ha hc ca XT1 c xc nh l axit

20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic. y l mt hp cht
c cu trc mi, ln u tin c phn lp t thin nhin.
Bng cc phng php tng t cho php xc nh cu trc ha hc ca tt c cc
cht cn li. Ton b cu trc ha hc v hot tnh sinh hc ca cc cht c tng hp
di y:
1. Hp cht XT1: axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17triynoic (cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:
6

7
9

COOH
1

17
15

11

Br
19

- Cht kt tinh mu trng

- Cng thc phn t: C20H21BrO2, M = 372
- Ph khi lng: m/z 394.8/396.8 [M+Na]+ vi t l 1/0,98 v cng tn hiu
- Hot tnh khng vi sinh vt kim nh: khng mnh ba chng vi sinh vt kim nh l:
trc khun m xanh Pseudomonas aeruginosa, khng t cu vng Staphylococcus
aureus v khng nm si Fusarium oxysporum vi gi tr MIC thp l 50 g/ml.
2. Hp cht XT2: axit 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19-triynoic (cht
mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:
8

COOH

11

19
13

17

22

Br

21

- Cht kt tinh mu trng

- Cng thc phn t: C22H27O2Br, M = 402
- Ph khi lng: m/z 424.9/426.9 [M+Na]+ vi t l 1/0,98 v cng tn hiu
- Hot tnh khng vi sinh vt kim nh: khng mnh ba chng vi sinh vt kim nh l:
trc khun m xanh Pseudomonas aeruginosa, khng t cu vng Staphylococcus
aureus v khng nm si Fusarium oxysporum vi gi tr MIC thp l 50 g/ml.
3. Hp cht XT3: cholesterol
- Cu trc ha hc:
21

20

21

24

25

18
17
19

26

1
8
3

HO

80

- Tinh th hnh kim mu trng:

mp 148-149oC
- Cng thc phn t: C27H46O,
M=386
- c phn lp t loi: Xestospongia
testudinaria

4. Hp cht XT4: 5,8-epidioxycholest-6-en-3-ol

- Cu trc ha hc:
22

21
20

18

25
27

17

11

13

19
1

26

9
8

10
3

15

HO

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng KB: IC50 = 2,0 g/ml
Dng FL: IC50 = 3,93 g/ml
Dng Hep-G2: IC50 = 2,4 g/ml

- Tinh th hnh kim mu trng: mp 102-105oC

- Cng thc phn t: C27H44O3, M = 416
- Ph khi lng: m/z: 417.0 [M+H]+, 439.0 [M
+Na]+
5. Hp cht XT5: cholest-7-en-3-on
- Cu trc ha hc:
21

22

18
12

11

19

20

1
10

- Tinh th hnh kim mu trng:

mp 147-148oC
- Cng thc phn t: C27H44O, M = 384
- Ph khi lng: m/z 385,2 [M+H]+

26
25

23

27

17

13

16
15

14
2

24

8
7

6. Hp cht XT6: saringosterol

- Cu trc ha hc:

- Tinh th hnh kim mu trng:

29

mp 160-161oC

28
21

24
20

18

25

OH

17

11

- Cng thc phn t: C29H49O2, M = 428

- Ph khi lng: m/z 411,1 [M-H2O+H]+,
393,0 [M-2H2O+H]+
- Hot tnh khng vi khun lao:
M. turbeculosis: MIC = 0,25 g/ml

26

27

13

19
1

14
8

10
3

HO

7. Hp cht XT7: thimidine

- Cu trc ha hc:

- Tinh th hnh kim mu trng:

mp 186-187oC
- Cng thc phn t: C10H14N2O5, M = 242
- Ph khi lng: m/z 264,7 [M+Na]+,
240.8 [M-H]-

CH3

HN3

5'

HOCH2
O
1'

4'
3'

2'

OH

81

8. Hp cht XT8: thimine

- Cu trc ha hc:

- Tinh th hnh kim mu trng: mp 326oC

- Cng thc phn t: C5H6N2O2, M = 126
- Ph khi lng: m/z 126,8 [M+H]+,
148,7 [M+Na]+, 124,7 [M-H]-

HN 3

2
1

CH3

N
H

9. Hp cht XT9: chimyl alcohol

- Cu trc ha hc:
1

- Tinh th mu trng: mp 64-65oC

- Cng thc phn t: C19H40O3, M=316
- Ph khi lng: m/z: 317,1 [M+H]+

OH

HO

1'

16'

CH2(CH2)14CH3

10. Hp cht XT10: batyl alcohol (batilol)

- Cu trc ha hc:
1

- Tinh th mu trng: mp 70-71oC

- Cng thc phn t: C21H44O3, M = 344
- Ph khi lng: m/z 345,1 [M+H]+

OH

HO

1'

18'

CH2(CH2)16CH3

Kt qu th nghim hot tnh khng vi sinh vt kim nh cho thy, ch c hp

cht XT1 v XT2 c biu hin hot tnh. Chng u khng mnh ba chng vi sinh vt
kim nh l: trc khun m xanh Pseudomonas aeruginosa, khng t cu vng
Staphylococcus aureus v khng nm si Fusarium oxysporum vi gi tr MIC thp l
50 g/ml.
Hin nay, nhiu loi thuc c tc dng iu tr cc bnh nhim khun v ang
c nghin cu v ng dng song hu ht cc thuc ny u gp kh khn trong vic
iu tr cc bnh nhim khun do trc khun m xanh Pseudomonas aeruginosa (thuc
nhm vi khun Gram (-) l vi khun gy khng thuc khng sinh mnh v khng hu
ht cc thuc khng sinh th h c) v t cu vng Staphylococcus aureus (thng gy
nhim trng v bi nhim nng ngoi da cng nh mt s c quan v thng c mt
trong cc trng hp nhim trng mu v cng l chng khng thuc vi nhiu thuc
khng sinh). Cc hp cht XT1 v XT2 c hot tnh mnh khng trc khun m xanh
Pseudomonas aeruginosa, khng t cu vng Staphylococcus aureus v khng nm si
Fusarium oxysporum. iu ny to c s khoa hc vng chc cho cc nghin cu ng
dng tip theo nhm to ra cc dc phm c tc dng phng v iu tr cc bnh nhim
khun da trn vic khai thc ngun dc liu bin qu gi, sn c trong nc vo phc
v cuc sng.
Ngoi ra, tuy khng c biu hin hot tnh trn 6 chng vi sinh vt kim nh
nghin cu, hp cht XT6 (saringosterol) li c hot tnh khng vi khun lao
Mycobacterium tuberculosis rt mnh vi gi tr nng c ch ti thiu (MIC) l 0,25
g/ml v y l gi tr MIC nh nht cho cc hp cht thin nhin c chit tch t
thc vt tnh n nm 2001. Gi tr ny tng ng vi gi tr MIC ca thuc cha lao
rifampin. iu ny chng t XT6 c kh nng khng vi khun lao vi tnh c hiu rt
cao. Cc nghin cu cng b cn chng minh hp cht ny c c tnh rt thp
[Gerald A. W., Scott G. F., Gloria M., Joseph J. H., Wiliam M. M., and Barbara N. T., Journal of Natural
82

Products, 2001, 64, 1463-1464]. Theo tc gi Brent R. C. [Brent R. C., Antimycobacterial natural
products, Natural Products Report, 2003, 20, 535-557.], vi lp cht steroit tnh n nm 2003,

ch c 3 hp cht l axit fusidic, saringosterol v endoperoxide c hot tnh khng vi

khun lao vi gi tr MIC 4 g/ml. iu ny t c s khoa hc cho hng nghin cu
su rng hn v hp cht XT6 cng nh loi hi min Xestospongia testudinaria theo
nh hng hot tnh khng vi khun lao nhm nghin cu v pht trin cc ch phm
thuc tr bnh lao th h mi, phc v cng tc chm sc sc khe cng ng.
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc loi hi min
Xestospongia testudinaria, chng ti bin son v ng ti 02 cng trnh trn tp ch chuyn ngnh
quc gia v 01 bng c quyn sng ch ti Cc S hu Tr tu Vit Nam l:
(1) Nguyn Xun Cng, Trn Anh Tun, Nguyn Thu Phng, V Vn Thnh, Phan Vn Kim v Chu
Vn Minh, Nghin cu thnh phn ha hc ca loi hi min Xestospongia testudinaria thu thp ti
Vit Nam. Tp ch Khoa hc v Cng ngh, 2007, 45(3), 43-50
(2) Chu Vn Minh, Phan Vn Kim v Nguyn Xun Cng, Two new brominated fatty acids from the
sponge Xestospongia testudinaria living in Vietnamese sea. Advance in Natural Science, 2008, 9(2),
167-171.
(3) Chu Vn Minh, Nguyn Xun Cng, Phan Vn Kim, L Mai Hng v Cng Thung, Hp cht
axt 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic v phng php chit hp cht
ny t loi hi min xestospongia testudinaria. S hu Tr tu Vit Nam, 2008, s n 1-2007-01616
SC ngy 8/8/2007 ang thm nh ni dung).

V.2. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi

hi min Gellius varius (chanlinidae)

- Tn Vit Nam: Hi min cnh

xanh
- Tn khoa hc: Gellius varius
- Thi gian thu mu: thng 01
nm 2007
- a im thu mu: Qung
Ninh
- Ngi nh loi: TS. Cng
Thung v cc cng s, Vin Ti
nguyn v Mi trng Bin,
VAST

Phn lp v tinh ch cc hp cht:

Mu hi min ti (3 kg) c xay nh, sau chit vi metanol ba ln, mi ln
72 gi. Cc dch chit metanol c gp li v ct loi dung mi di p sut gim thu,
ha vo nc ri tin hnh loi mui trn ct Dianion vi dung mi ra gii l nc.
Dng MeOH ra gii thu hi cht thu c 15 gam dch c. Dch c ny c ho
tan vo 01 lt nc ct sau c chit ln lt bng n-hexan, cloroform, v BuOH
(mi ln 02 lit) thu c cc cn dch tng ng: hexan (2 g), cloroform (4 g), BuOH (6
g) v nc (3 g) (s V.2.a.).

83

Hi min Gellius varius ti

(3 kg)
- Chit MeOH, ct loi dung mi di p
sut gim
- Loi mui bng cch chy qua ct
Dianion vi dung mi ra gii l nc ct,
sau thu hi cht bng MeOH.
B sung nc

GM - Cn chit MeOH (15g)

B sung n-hexan

n-Hexan: nc 1/1

Lp nc
B sung CHCl3

GH - Cn n-hexan (2 g)

CHCl3: nc 1/1

Lp nc
B sung BuOH

GC - Cn CHCl3 (4 g)

BuOH: nc 1/1
lp nc

GB - Cn BuOH (6 g)

GN - Cn nc (3 g)

S V.2.a: S chit phn on mu Hi min

GH - (2 g)
Silica gel CC
Hexan/Axeton 100/0-0/100

GH1 (250mg)

GH2 (375mg)

GH3 (225mg)

Silica gel CC
n-hexan/Etylaxetat
40/1
HM-8
(10mg)

GH4 (600mg)

GH5 (550g)

Silica gel CC
n-hexan/axeton/ EtOAc:
50/1/1

HM-9
(100mg)

S V.2. b: S phn lp cc hp cht t cn chit n-hexan

84

HM-10
(5mg)

GC (4 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

GC1 (1g)

GC2 (600mg)

GC4 (200mg)

GC5 (900mg)

Silica gel CC
n-hexan/CHCl3/axeton 4/1/0,1

Silica gel CC
n-hexan/axeton: 7/1

HM-4
(7mg)

GC3 (1,3g)

HM-5
(12mg)

Silica gel CC
CHCl3/MeOH
20/1

HM-7
(9mg)

HM-6
(8mg)

S V.2. c: S phn lp cc hp cht t cn chit cloroform

Cn chit n-hexan: GH (2 g) c tin hnh phn tch trn sc k ct lp li nhiu
ln vi cc h dung mi thch hp thu c cc hp cht HM-8, HM-9 v HM-10. Cn
chit clorofrom: GC (4 g) c tin hnh chy ct th trn sc k ct pha thng
gradient nng metanol trong cloroform thu c 5 phn on nh hn l GC1-5. Cc
phn on GC1, GC3 v GC5 c tip tc phn tch trn sc k ct pha thng vi h
dung mi ra gii thch hp thu c cc hp cht HM-4, HM-5, HM-6 v HM-7 nh
c ch ra trn. Bng cc phng php tng t, cc hp cht HM-1, HM-2, v HM-3
c phn lp t cn chit BuOH (s V.2.b-d).

GB - (6 g)
Silica gel CC
Cloroform/MeOH/H2O: 5/1/0 - 2/1/0.2

GB1 (1,0 g)

GB2 (2,5 g)

Silica gel CC
CHCl3/MeOH/H2O
5/1/0,1
HM-1
(12mg)

GB3 (1,5 g)

GB4 (0,6 g)

Silica gel CC
CHCl3/MeOH/H2O
4/1/0,1
HM-2
(90mg)

GB5 (1,4 g)

Silica gel CC
CHCl3/MeOH/H2O
3/1/0,2
HM-3
(15mg)

S V.2. d: S phn lp cc hp cht t cn chit butanol

85

Xc nh cu trc ha hc v hot tnh sinh hc ca cc hp cht:

1. Hp cht HM1: 1-O-palmitoyl-3-O-[-D-galactopyranosyl-(16)--D-galactopyranosyl]-sn-glycerol
- Cu trc ha hc:
- Hot tnh sinh hc:
6"
HO
+ Hot tnh gy c t bo
5"
O
OH
Dng KB: IC50 = 3,51 g/ml
1"
OH
4"
Dng Hep-G2: IC50 = 3,15
6'
O
2"
3"
g/ml
3
5'
O
O CH
OH OH
+ Hot tnh khng VSV kim nh
2
OH
4'
OH
1' HC
P. aeruginosa: MIC = 150
1'"
16'"
2'
3'
1
CH O C (CH ) CH
g/ml
OH
O
S. aureus: MIC = 200 g/ml
- Cht bt v nh hnh mu trng
- Cng thc phn t: C31H58O14, M = 654
- Ph khi lng: m/z 653,3 [M-H]+, 677,3 [M+Na]+,
2

2 14

515,2 [M-162+Na]+, 689,4 [M+2H2O-H]+

2. Hp cht HM2: (2S)1,2-di-O-palmitoyl-3-O-(6-sulfo--D-quinovopyranosyl)

glycerol
- Cu trc ha hc:
- Hot tnh sinh hc:
6'
+ Hot tnh gy c t bo
CH2SO3H
5'
Dng FL: IC50 = 4,06 g/ml
O
1'
4'
Dng Hep-G2: IC50 = 2,89 g/ml
OH
3
2'
+
Hot tnh khng VSV kim nh
OH
O CH2
3'
16'"
1'"
P. aeruginosa: MIC = 100 g/ml
2
OH
HC O C (CH2)14CH3
B. subtilis: MIC = 100 g/ml
O 1"
16"
1
CH2 O C (CH2)14CH3
F. oxysporum: MIC = 50 g/ml
O
S. cerevisiae: MIC = 200 g/ml
- Cht bt v nh hnh mu trng
- Cng thc phn t: C41H78O12S, M = 794
- Ph khi lng: m/z 793,6 [M-H]+
3. Hp cht HM3: mui natri ca (2S)1-O-hexadecanoyl-3-O-(6-sulfo--Dquinovopyranosyl) glycerol
- Cu trc ha hc:
- Hot tnh sinh hc:
6'
+ Hot tnh gy c t bo
CH2SO3Na
5'
Dng Hep-G2: IC50 = 2,71 g/ml
O
1'
4'
+ Hot tnh khng VSV kim nh
OH
3
2'
OH
O CH2
E. coli: MIC = 200 g/ml
3'
2
OH
B. subtilis: MIC = 200 g/ml
HC OH
S. cerevisiae: MIC = 200 g/ml
1"
16"
1
CH2 O

(CH2)14CH3

- Cht bt v nh hnh mu trng

- Cng thc phn t: C25H47NaO11S, M = 578
- Ph khi lng: m/z 555,3 [M-Na]+,
601,2 [M+Na]+

86

4. Hp cht HM4: cholesterol

- Cu trc ha hc:
21

20

21

24

25

18
17

26

19
1

- Tinh th hnh kim mu trng:

mp 148-149oC
- Cng thc phn t: C27H46O,
M=386

8
3

HO

5. Hp cht HM5: 5,8-epidioxycholest-6-en-3-ol

- Cu trc ha hc:
22

21
18

26

25
27

17

11

13

19
1

20

9
8

10
3

7 O

HO

15

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng KB: IC50 = 2,0 g/ml
Dng FL: IC50 = 3,93 g/ml
Dng Hep-G2: IC50 = 2,4 g/ml

- Tinh th hnh kim mu trng: mp 102-105oC

- Cng thc phn t: C27H44O3, M = 416
- Ph khi lng: m/z: 417,0 [M+H]+, 439,0 [M
+Na]+
6. Hp cht HM6: cholest-7-ene-3,5,6-triol
- Cu trc ha hc:
21

18

27

10
5

HO

25

13

19

26

20

17

11
1

22

15

- Tinh th hnh kim mu trng

- Cng thc phn t: C27H46O3, M =
418
- Ph khi lng: m/z 401,2 [MH2O+H]+, 383,2 [M-2H2O+H]+,
365,2 [M-3H2O+H]+

OH
OH

7. Hp cht HM7: 6-methoxycholest-7-ene-3,5-diol

- Cu trc ha hc:
21
22
26
- Tinh th hnh kim mu trng
20
18
25
- Cng thc phn t: C28H48O3, M =
27
17
432
11
13
19
- Ph khi lng: m/z 415,3 [M1

10

HO

5
4

H2O+H]+, 397,2 [M-2H2O+H]+, 383,3

[M-2H2O-Me+H]+, 365,3 [M-2H2O-MeH2O+H]+, 431,1 [M-H]+

15

OH
OCH3

87

8. Hp cht HM8: metyl este ca axit (5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic

- Cu trc ha hc:
O

11

20

17

14

- Cht du khng mu
- Cng thc phn t: C21H32O2, M = 316
- Ph khi lng: m/z 317,1 [M+H]+, 315,0 [M-H]+
9. Hp cht HM9: trans-phytol
- Cu trc ha hc:
17

18
11

15

19

20

OH

16

- Cht du khng mu
- Cng thc phn t: C20H40O, M = 296
10. Hp cht HM10: glycerol 1-(5Z,8Z,11Z,14Z-eicosatetraenoate)
- Cu trc ha hc:
3

HO
O
5'

8'

OH
2

1'

O
20'
11'

14'

- Cht du khng mu
- Cng thc phn t: C23H38O4 (M=378)
- Ph khi lng: m/z 379,2 [M+H]+, 361,2 [M-H2O+H]+, 347,2 [M-H2O-CH2+H]+, 329,1 [MH2O-CH2-H2O+H]+, 287,2 [M-Glycerol+H]+, 377,1 [M-H]+

Hp cht HM5 th hin hot tnh trn c 3 dng t bo th nghim. Hp cht

HM1 v HM2 th hin hot tnh 2/3 dng t bo th nghim. Hp cht HM3 th hin
hot tnh 1/3 dng t bo th nghim. Cc kt qu nghin cu trn cho thy s ph hp
vi cc ti liu cng b. Cc hp cht glycolipit th hin nhiu hot tnh qu bu
trong c hot tnh gy c t bo. C th l hp cht HM2 th hin hot tnh gy c
t bo i vi cc dng t bo u P388 v HL60. Ngoi ra n cn th hin hot tnh gy
c t bo i vi cc dng t bo u ngi l MCF7 v A549 (Shu-Hua Qi, et al., Chem.
Pharm. Bull., 2004, 52(8), 986-988). Gn y M c mt s bng c quyn sng ch
c cp v kh nng ng dng lm thuc ca cc dn xut thuc lp cht sulfonoglycolipit (Yamazaki T., et al., United States Patent, 2004, No. US 6,759,522 B2). Theo cc tc gi
ny, lp cht ny thng th hin hot tnh gy c t bo v l cc cht c ch enzym
tng hp DNA (DNA-polymerase). Cc hp cht ny c phn lp vi hm lng kh
cao t loi hi min G. varius sinh sng rt ph bin ti cc vng bin Vit Nam. iu
ny gi m cho nhng nghin cu su hn v hp cht ny theo hng ng dng lm
dc liu.

88

Kt qu th nghim hot tnh khng vi sinh vt kim nh cho thy, hp cht HM1
khng 03 chng vi sinh vt kim nh l P. aeruginosa v S. aureus, hp cht HM2
khng 04 chng l P. aeruginosa, B. subtilis, F. oxysporum v S. cerevisiae, HM3
khng 03 chng l E. coli, B. subtilis, v S. cerevisiae. Hp cht HM2 v HM3, thuc
nhm cht sulfuno glycolipit, c cu trc c th vi s c mt ca d t lu hunh (S)
trong phn t khng nhng gi nhiu chc nng quan trng trong c th sinh vt m cn
th hin nhiu hot tnh qu bu. Chng c phn lp vi hm lng cao trong loi hi
min G. varius sinh sng ph bin ti cc vng bin Vit Nam. iu ny t c s khoa
hc cho cc nghin cu tip theo nhm to ra cc dc phm mi trong iu tr cc bnh
him ngho ng thi s dng mt cch hp l v c hiu qu ngun dc liu bin v
cng qu gi ca t nc.
T cc kt qu nghin cu v thnh phn ha hc v hot tnh sinh hc loi hi min Gellius
varius, chng ti bin son v ng 02 cng trnh trn tp ch chuyn ngnh quc gia l:
(1) Nguyn Xun Cng, Phan Vn Kim v Chu Vn Minh, Cc hp cht glycolipit t loi hi min
cnh xanh Gellius varius sinh sng ti vng bin Vit Nam. Tp ch Khoa hc v Cng ngh, 2007,
45(2), 41-47.
(2) Nguyn Xun Cng, Nguyn Hi ng, Cng Thung, Trn Thu Hng, Nguyn Th Kim Thy,
Phan Vn Kim v Chu Vn Minh, Isoprenoids from the sponge Gellius varius living in Vietnamese
sea. Tp ch Ha hc, 2007, 45(6), 776-780.
Cc kt qu trn c s dng trong 01 Lun n Thc s bo v thnh cng (ThS. Nguyn Th Kim
Thy)

V.3. Kt qu nghin cu ho hc v hot tnh sinh hc ca loi

hi min Ianthella sp. (Ianthellidae)

- Tn Vit Nam: Hi min

- Tn khoa hc: Ianthella sp.
- Thi gian thu mu: thng 03
nm 2007
- a im thu mu: Nam Yt,
Qun o Trng Sa, tnh Khnh
Ha
- Ngi nh loi: TS. Cng
Thung v cc cng s, Vin Ti
nguyn v Mi trng Bin, VAST

Phn lp v tinh ch cc hp cht:

Hi min Ianthella sp. (10 kg ti) c ra sch, xay nh v chit bng metanol
(3 ln) bng thit b chit siu m. Dch metanol ca cc ln chit c lc, gom li v
ct loi dung mi nhit khng qu 60oC di p sut gim thu c 120 g dch cn
dch metanol. Cn dch ny c ha vo nc v chit phn b ln lt vi cc dung
mi hexan v etyl axetat theo t l 1/1 (3 ln, mi ln 6 lit) thu c cc cn dch tng
ng l hexan 10 g (k hiu: IH), EtOAc 9.3 g (k hiu IE) v nc 50 g (k hiu IN) (s
V.3.a).

89

Hi min Ianthella sp. ti

(10 kg)

- Chit MeOH, ct loi dung mi di p sut gim

B sung nc
B sung hexan

IM - Cn chit MeOH (70 g)

hexan: nc 1/1

Lp nc
B sung EtOAc

Cn hexan
IH (10 g)

EtOAc: nc 1/1

Lp nc

Cn nc
IN (50 g)

Cn EtOAc
IE (9,3 g)

S V.3.a: S chit phn on mu hi min

Cn chit EtOAc: IE (9.3 g) c tin hnh chy ct th trn sc k ct pha
thng gradient nng metanol trong cloroform thu c 4 phn on nh hn l IE1IE4. Cc phn on IE1, IE2 v IE3 c tip tc phn tch trn sc k ct pha thng
v pha o vi h dung mi ra gii thch hp thu c cc hp cht IS1 n IS5 (S
V.3.b-c).
IE (9.3 g)
Silica gel CC
CHCl3/MeOH 100/0-0/100

IE1 (0.5 g)

Silica gel CC
CM 30/1
IS1
90 mg

IE2 (1 g)

IE3 (1.8 g)

IE4 (6 g)

YMC RP-18 CC
MWA 5/1/3
IS3
7,5 mg

S V.3. b: S phn lp cc hp cht t cn chit EtOAc

90

IE3 (1.8 g)
Silica gel CC
HE 5/1

IE3A (0.2 g)

IE3B (0.6 g)

IE3C (1.0 g)

RP CC (YMC ODS-A)
MeOH-H2O-Acetone (5:1:3)
IS4
10 mg

IE3C1 (100mg)

RP CC (YMC ODS-A))
MeOH-H2O-Acetone (5:1:3)

IE3C2 (0,5 g)

IE3C3 (0,4 g)

CC (silica gel,)
n-hexane:EtOAc=3/2

Kt tinh li
trong axeton
IS2
12 mg

IS5
(10mg)

S V.3.c: S phn lp cc hp cht t cn chit cloroform

Xc nh cu trc ha hc v hot tnh sinh hc ca cc cht:
S dng kt hp nhiu phng php ph hin i, chng ti xc nh c cu
trc ha hc ca 05 hp cht steroit c cu trc vng propan mch nhnh (03 cht
mi) t loi hi min Ianthella sp. Cc hp cht ny c tin hnh nh gi hot
tnh gy c t bo. Ton b kt qu c tng hp di y:
1. Hp cht IS1: petrosterol
- Cu trc ha hc:

29

21
18

19

12
11

13
14

2
3

HO

10
5

22
20

26

24

28
23

25
27

17
16
15

8
7

- Cht bt mu trng
- Cng thc phn t: C29H48O, M = 412
91

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 5,24 M
Dng SK-Hep-1: IC50 = 7,31 M
Dng Hela: IC50 = 15,63 M

2. Hp cht IS2: iansterol A (cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
29
- Ph khi lng phn gii cao HRFAB-MS: m/z 425,3397 [M+H]+
21
22
26
24
(tnh ton l thuyt cho C29H45O2 l
28
18
20
25
23
425,3375)
27
12
- Hot tnh sinh hc:
17
11
13
19
16
+ Hot tnh gy c t bo
14
15
Dng MCF-7: IC50 = 3,74 M
9
1
8
10
2
Dng SK-Hep-1: IC50 = 9,71 M
7
5
3
Dng Hela: IC50 = 12,84 M
4
6
O

- Cht bt mu trng
- Cng thc phn t: C29H44O2 (M = 424)
- Ph khi lng: m/z 425 [M+H]+
3. Hp cht IS3: iansterol B (cht mi, ln u tin c phn lp t thin nhin)
- Cu trc ha hc:
29
- Ph khi lng phn gii cao HR21
EI-MS: m/z 428,3656 [M+H]+ (tnh
22
26
24
28
ton l thuyt cho C29H48O2 l
18
20
25
23
428,3654)
27
12
17
- Hot tnh sinh hc:
11
13
19
16
+ Hot tnh gy c t bo
14
15
9
1
Dng MCF-7: IC50 = 17,36 M
8
2
10
Dng SK-Hep-1: IC50 = 13,84 M
7
5
3
4
6
Dng Hela: IC50 = 18,67 M
HO
O

- Cht bt mu trng
- Cng thc phn t: C29H48O2 (M = 428)
- Ph khi lng: m/z 429 [M+H]+
4. Hp cht IS4: aragusterol B
- Cu trc ha hc:

29

21

HO

18

12
19

11

13
14

2
3

20

26

24

28
23

25

17

27

16
15

10
5

22

HO

7
6

- Cht bt mu trng
- Cng thc phn t: C29H48O3, M = 444
- c phn lp t loi: Ianthella sp.

92

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 12,83 M
Dng SK-Hep-1: IC50 = 18,53 M
Dng Hela: IC50 = 27,80 M

5. Hp cht IS5: iansterol C (cht mi, ln u tin c phn lp t thin nhin)

- Cu trc ha hc:
29
- Cht bt mu trng
21
26
22
- Cng thc phn t: C31H54O4, M =
24
HO
28
18
HO
490
20
25
23
- Ph khi lng: m/z 461 [M - CH2O +
27
12
19

11

13
14

H3CO

2
3

9
5

- Hot tnh sinh hc:

+ Hot tnh gy c t bo
Dng MCF-7: IC50 = 16,95 M
Dng SK-Hep-1: IC50 = 25,32 M
Dng Hela: IC50 = 24,61 M

15

10

H]+

17
16

7
6

OCH3

Tt c cc hp cht phn lp c t loi hi min Ianthella sp. c tin hnh

nh gi hot tnh gy c t bo. Kt qu th nghim cho thy, tt c cc hp cht u
th hin hot tnh mnh chng li 3 dng t bo ung th ngi th nghim l MCF-7
(v), SK-Hep-1 (gan) v Hela (c t cung). Kt qu ny hon ton ph hp vi cc
nghin cu trc y. Nhiu cng trnh cng b u khng nh rng cc hp cht
sterol c cu trc b khung tng t u th hin hot tnh gy c t bo rt mnh. Cc
tc nhn gy c t bo c s dng trong iu tr mt s loi ung th nh ung th
phi, ung th v... v c coi l mt liu php chnh trong cc tr liu ung th bng
phng php ha hc. Cc tc nhn ny biu hin c tnh bng cch tiu dit cc t bo
ung th theo cc c ch khc nhau, ch yu l can thip vo qu trnh phn chia t bo,
c bit qu trnh sao chp DNA. Cc steroit c phn lp t loi hi min Ianthella sp.
c cu trc c o v th hin hot tnh gy c t bo cao i vi cc dng t bo ung
th c th nghim, iu ny gi m cho cc nghin cu tip theo theo hng ng
dng lm dc liu.
chng VI: Tng hp cc hp cht phn lp c trong
khun kh ti KC09.09/06-10 v hot tnh sinh hc ca
chng
Trong khun kh ti KC09.09/06-10, chng ti tin hnh phn lp v xc
nh cu trc c 51 hp cht. Trong 18 hp cht c cu trc mi ln u tin
c phn lp t thin nhin. Ton b cc hp cht phn lp c c tin hnh
nh gi hot tnh: gy c t bo, chng xy ha v khng vi sinh vt kim nh. Ngoi
ra, thng qua hp tc quc t, chng ti tin hnh nh gi hot tnh chng long
xng thng qua cc c ch kch thch s bit ha ca nguyn bo xng v khng yu
t phin m NF-B ca mt s hp cht.
Trong chng ny, chng ti tng hp cu trc v hot tnh ca ton b cc hp
cht phn lp c v chia thnh 09 nhm cht bao gm: ditecpen dng cembranoit,
9,11-secosterol, tritecpen saponin (cc holothurin), cerebrosit v ceramit, axit bo khng
no b brm ha, steroit c vng propan mch nhnh, sterol mang nhiu nhm hydroxy
trong phn t (polyhydroxylated sterols), glycolipit, v cc lp cht khc.
T l cc cht mi trong tng s cht phn lp c thuc cc nhm cht
ditecpen dng cembranoit, 9,11-secosteroit, holothurin, axit bo a ni i - ni ba b
brm ha, steroit c cu trc vng propan mch nhnh, cerebrosit v ceramit l rt
cao. y u l cc lp cht c cu trc c th v gi nhiu chc nng sinh hc quan
trng trong c th sinh vt cng nh th hin nhiu hot tnh qu bu. Cc cembranoit
gi vai tr phng v ha hc ca cc sinh vt chng li k th. Chng thng th hin
nhiu hot tnh sinh hc qu bu nh bo v thn kinh, khng vi sinh vt, khng vim v
93

khng ung th. Cc 9,11-secosterol tn ti vi s lng khng nhiu trong t nhin,

chng thng th hin hot tnh gy c t bo v chng tng sinh rt nhiu dng t
bo ung th khc nhau nh C6 (t bo lioma chut), J774 (bch cu n nhn to v i
thc bo chut), LnCap (t bo ung th tuyn tin lit), Calu-3 (ung th phi).... Cc
holothurin l cc thng c ph hot tnh rng nh: gy c t bo, khng nm, tan
mu, iu bin min dch... Cc holothurin th hin nhiu hot tnh qu bu nn c gi
tr rt cao. n c nh hp ct holothurin A c hng Sigma rao bn vi gi 5000
USD/1g. V phng din sinh hc, mt s cerebrosit c chng minh gi vai tr h
tr cu trc v xc nh kt cu ca cc mng t bo, chng hot ng nh cc cht iu
tit ca nhiu qu trnh sinh hc nh qu trnh hot ha, qu trnh dnh kt t bo, trao
i thng tin ni bo v pht trin ca t bo. Hn na, cc cerebrosit trong mng t bo
gi vai tr sinh hc quan trng l cc khng nguyn b mt v cc th th. c bit,
ngy cng c nhiu bng chng ch ra rng cc cerebrosit c cc chc nng sinh hc rt
a dng, hu ht cc chc nng ny u lin quan n c tnh lng cc ca phn t.
Chng hn nh, nhn t pht trin thn kinh (NGF - Nerve growth factor), mt i phn
t sinh hc l nhn t u tin c xc nh r nht, chng iu tit s pht trin, bit
ha v sng st ca cc nron v c coi l mt ng vin nng k pht trin cc
thuc tr bnh Alzheimer (bnh qun). Tuy nhin, phn t NGF qu ln v mt kch
thc v qu a nc i qua cc vch ngn mu no, m li l con ng an ton
duy nht a cc NGF vo trong no tht. Do , cc cht phn t lng thp c tc
dng tng t cc NGF (chng hn nh cc cerebrosit) l cc ng vin y ha hn cho
vic tr bnh Alzheimer. Cc hp cht axit mch di a ni i v ni ba b brm ha
thng c hot tnh khng sinh v gy c t bo. c bit, mt s hp cht th hin
hot tnh c ch enzym HIV protease, mt enzym thit yu cho s ti bn ca HIV.
Thm vo , chng cn c hot tnh km hm tc nhn i khng lin kt vi th th
Adenosine. Khi th th ny c hot ha s mang n cc hiu qu tch cc nh gim
nhp tim, bo v no v tiu m.
Cc nghin cu trong khun kh ti cho thy, cc hp cht cembranoit v 9,11secosteroit cn th hin hot tnh chng long xng thng qua cc c ch kch thch s
pht trin v bit ha ca nguyn bo xng. Kt qu ny cho thy cc hp cht hot
ng nh mt tc nhn iu bin chn lc (SERM) v iu bin qu trnh ri lon xng
ph n thi k mn kinh. Cc hp cht ny c phn lp vi hm lng kh cao, iu
ny gi m cho nhng nghin cu su hn v cc hp cht ny theo hng ng dng
trong vic phng v iu tr bnh long xng, mt cn bnh rt ph bin hin nay (c
bit l trong cng ng ngi cao tui). Ngoi ra, cc hp cht 9,11-secosterol, steroit
mang nhiu nhm hydroxy, cerebrosit v ceramit cn th hin hot tnh khng yu t
phin m NF-B. Yu t ny iu tit s biu hin ca nhiu gen tham gia vo cc p
ng min dch v vim. N lm tng s biu hin ca cc gen quy nh nhiu cytokine,
cc enzym v cc phn t dnh kt trong cc bnh vim mn tnh. NF-B c hot ha
bi nhiu yu t khc nhau (virus, cc cht xy ha v cc khng nguyn) lm tng p
ng vim. S hot ha ny li dn ti s biu hin phi hp ca nhiu gen m ha cc
protein (nh cc cytokine, chemokine, enzym v cc phn t dnh kt) tham gia vo s
tng hp cht trung gian v ko di thm p ng vim. V vy, NF-B l mt ch hin
nhin cho cc phng php iu tr khng vim mi. Glucococticoit l cc cht c ch
NF-B hu hiu nhng chng c nhiu tc dng ph ti ni tit v chuyn ha khi c
s dng vi liu php iu tr ton thn. Cc tc dng ph ny s khng xy ra khi s
dng cc cht c ch NF-B c hiu hn. V vy, vic tm kim v nghin cu k lng
cc hp cht thin nhin c tc dng c ch c hiu NF-B v ang thu ht mnh
m s quan tm v u t nghin cu ca cc nh khoa hc trn ton th gii.
94

Nh vy, cc kt qu nghin cu thnh phn ha hc v hot tnh sinh hc t

c trong ti mang ngha khoa hc rt cao (c bit l trong lnh vc ha hc cc
hp cht thin nhin bin, mt lnh vc nghin cu cn rt non tr Vit Nam) v m ra
nhiu hng nghin cu ng dng pht trin cc dc t mi t ngun dc liu bin
di do ca t nc trong tng lai. Cc kt qu c ng trn 17 bi bo khoa
hc trong c 11 bi trn cc tp ch chuyn ngnh quc gia v 06 bi bo quc t (05
bi ng v 01 bi gi ng). Ngoi ra ti ng k c 02 bng c quyn
sng ch ti cc s hu tr tu Vit Nam (01 bng c cp). iu ny s gp phn
a cc nghin cu v ha hp cht thin nhin bin ca nc ta ln ngang tm cc nc
trong khu vc cng nh quc t ng thi bo h c cc nghin cu ca t nc.

C. Cc nghin cu thm d v rong, to v vi sinh vt bin

Chng VII. Nghin cu thm d kh nng khng vim t
rong, to bin vit nam.
VII.1. M u
Phn ny trnh by s lc v vim, mt s hot cht khng vim in hnh v c
ch hot ng ca chng.

VII.2. KT QU V THO LUN

VII.2.1.Thu thp v nh tn s b 19 loi Rong to bin Vit Nam
thu thp v nh tn khoa hc c 19 loi rong to mt s tnh pha Bc
(Hi Phng), min trung (Tha Thin, Hu) v min Nam (Nha Trang, Khnh Ho,
Ninh Thun), Vit Nam. Danh sch cc loi nu phn trn.
VII.2.2. Hot tnh ca cht khng vim c chit t Undaria pinnatifida bng cc
loi dung mi khc nhau
Kt qu phn tch hot tnh khng vim t Undaria pinnatifida c chit bng cc
loi dung mi khc nhau c ch ra trn bng VII.2.2.
Bng VII.2.2. Hot tnh ca dch chit khng vim c chit t Undaria pinnatifida
bng cc loi dung mi khc nhau
Dung mi

Gi tr ph n

Acetonitrile
Acetone
Benzene
Chloroform
Cyclohexane
Ethyl acetate
Ethyl alcohol
te
Hexan
Metanol
Methylene chloride
Nc kh trng
Nc si
i chng

0,03 0,00
0,17 0,02
0,03 0,00
0,03 0,00
0,13 0,00
0,12 0,00
0,27 0,00
0,12 0,02
0,62 0,02
0,12 0,00
0,03 0,00
0,70 0,12
0,80 0,00
0,81 0,04

Phn trm c
ch
96
79
96
96
84
85
67
85
23
85
96
14
1
0

Gi tr ban
0,010,00
0,03 0,00
0,04 0,00

Phn trm c
ch
96
89
85

0,06 0,01

78

0,14 0,04

48

0,10 0,00
0,06 0,00

63
78

0,17 0,00
0,16 0,02
0,27 0,01

37
4
0

Ghi ch: *: khng xc nh; i chng: khng c bi dch chit c hot tnh khng vim. - Mi gi tr
i din trung bnh S.D. ca t 6 n 7 con chut. Sai khc c ngha thng k P<0,05

95

Kt qu trong bng VII.2.2 cho thy, hot tnh khng vim trong cc dch chit t
U. pinnatifida vi cc dung mi khc nhau l khc nhau. Hot tnh khng vim cao nht
khi rong c chit vi acetonitrile, benzen v metanol. Tng ng vi phn trm c ch
ph n v ban trong dch chit vi acetonitrile l 96% v 96%; trong benzen l 96%
v 85%, v trong metanol l 85% v 78%, tng ng tng gi tr. Tuy nhin, tch mt
lng ln cht khng vim vi gi thnh r v cht lng tt phc v cho cc th
nghim tip theo chng ti chn dung mi tch chit l Metanol
VII.2.3. Sng lc hot tnh khng vim trong cc loi rong bin Vit Nam
Tt c cc loi rong nghin cu u c hot tnh khng vim. Cc loi rong bin
khc nhau th hot tnh khng vim cng khc nhau. Trong , Ulva reticulata v
Undaria pinnatifida cho hot tnh khng vim cao nht (chim trn 75% gi tr c ch
ph n v ban ), sau n cc loi Enteromopha linza v Sargassum (S. duplicatum,
S. polycystum, S. mcclurei, S. denticarpum, S. swartzii, S. binderi) (50-70% gi tr c ch
ph n v ban ). Hot tnh khng vim thp nht l trong cc loi Gracilaria arcuata,
G. eucheumoides v G. asiatica (nh hn 20% gi tr c ch ph n v ban ).
VII.2.4. nh hng ca thi gian iu tr vim bng dch chit t Undaria
pinnatifida ln kh nng khng vim tai chut
Sau 0,5 n 2 gi iu tr bng dch chit Undaria pinnatifida u c hot tnh c
ch ban . Tuy nhin, ch c phn trm c ch ban trong l chut c iu tr sau
1h l c s sai khc c ngha thng k khi so snh vi i chng. Phn trm c ch
ph n ca dch chit U. pinnatifida cc thi gian iu tr khc nhau cng khc nhau.
Kh nng c ch ph n ca dch chit loi ny t cao nht ti thi im sau 1 gi iu
tr (85,63%). Sau thi gian iu tr 2 gi, kh nng c ch ph n v ban trong dch
chit ny gim xung ng k. Do , chng ti chn thi gian iu tr vim bng dch
chit c hot cht khng vim t rong bin cho cc th nghim tip theo l sau 1 ting.
VII.2.5. nh hng ca cc nng dch chit trong cc loi rong bin nghin cu
ln kh nng khng vim tai chut
Da trn kt qu sng lc cc loi rong c hot tnh khng vim, chng ti chn
ra mt s loi nh l Undaria pinnatifida, Gracilaria blodgettii, Ulva reticulata,
Sargassum swartzii v S. polycystum nghin cu su hn v nh hng ca chng
cc nng dch chit khc nhau ln kh nng khng vim trn tai chut.
Kt qu cho thy phn trm c ch ca dch chit Undaria pinnatifida, Gracilaria
blodgettii, Ulva reticulata, Sargassum swartzii v S. polycystum cc nng khc
nhau cng khc nhau. Trong phn ln cc loi nghin cu gi tr ny tng dn theo nng
. Khi so snh dch chit thu c t cc loi rong nghin cu nng 0,08ug/ml,
chng ti thy rng dch chit ca Ulva reticulata v Undairia pinnatifida c hot tnh
khng vim cao nht (th hin phn trm gi tr c ch ban t n > 80% v c
ch ph n > 75%). Tip theo l hai loi Sargassum swartzii v S. polycystum c thu
ven b bin tnh Ngh An v do cng ty TNHH Vit i cung cp u c hot tnh
khng vim gn tng ng nhau (phn trm gi tr c ch ban 58, 83 v 55,25%;
c ch ph n - 60,14 v 55,06%, tng ng tng loi mt). Cn loi rong cu Tht - G.
blogettii c hot tnh khng vim thp nht (26,23 v 6,09%).
Trn c s cc s liu thu c trn, chng ti chn ra hai loi c hot tnh khng
vim cao v tr lng ln l Ulva reticulata (thu thp ti Nha Trang, Khnh Ha) v
Sargassum swartzii tch chit lng ln dch chit khng vim. Dch chit ca hai
loi ny sau , c s dng cho cc nghin cu su hn v th hot tnh khng sinh
trn cc chng vi sinh vt kim nh v khng vim trn chut (th c tnh cp, tc
dng gim au bng axit acetic, gim au mm nng, gim vim mn bng u ht, gy
vim cp mng bng bng carageenane).
96

VII.2.6. Kt qu th hot tnh vi sinh vt v nm trn cc chng vi sinh vt v nm

kim nh
Kt qu cho thy, dch chit ca loi Ulva reticulata v Sargassum swartzii c
hot tnh khng chng Bacillus subtilis (vi gi tr IC50 = 62,15 g/ml) v
Staphylococcus aureus (IC50= 56,53 g/ml), tng ng tng loi mt
VII.2.7. Th nghim c tnh cp
Sau khi cho chut ung dch chit Sargassum swartzii v Ulva reticulata n liu
cao nht (66g/kg), chng ti thy rng chut sau ung thuc th vn hot ng, n ung
v i li bnh thng, phn v nc tiu ca chut khng c g bt thng. Sau 72 gi,
khng c chut no cht. V khng th tng th tch theo ng ung hn c na (th
tch thuc ti a chut c th dung np) nn cha tnh c LD50 ca cao khng vim
Sargassum swartzii v Ulva reticulata trn chut nht trng theo ng ung.
VII.2.8. Tc dng gim au
* Theo phng php qun au axit acetic (Koster):
Kt qu cho thy, dch chit Sargassum swartzii c 2 liu thp v cao u lm
gim r rt s cn au ca chut so vi l i chng m cc thi im 5-10, 10-15,
15-20sau khi gy au cho chut bng axit axetic (P<0,05 - 0,001). Liu 500mg/kg c
tc dng tt hn v mnh hn so vi aspirin liu 100mg/kg. Dch chit Ulva reticulata
liu 500mg/kg cng lm gim s cn au ca chut v tng ng vi Aspirin liu
100mg/kg.
* Theo phng php gy au bng mm nng:
Kt qu cho thy, dch chit Sargassum swartzii v Ulva reticulata liu 500mg/kg
lm gim au r rt thng qua vic lm ko di thi gian tc dng phn ng vi nhit
khi so snh gia trc v sau khi dng thuc th (P < 0,05). Tc dng ca hai dch chit
ny yu hn morphin liu 10mg/kg.
VII.2.9. Tc dng chng vim cp
* Trn m hnh gy ph chn chut bng carrageenin:
Kt qu cho thy, dch chit Sargassum swartzii c 2 liu 175mg/kg v
350mg/kg c tc dng chng vim r rt thng qua lm gim th tch chn chut gy
ph bng carrageenin cc thi im 2, 6 v 24 gi (P< 0,05). Tc dng chng vim
ca dch chit t Sargassum swartzii (52,12% liu 175mg/kg v 45,84% liu
350mg/kg) mnh hn khi so snh vi Indometacin liu 25mg/kg (33,145%) thi im
24 gi sau gy vim. Dch chit Ulva reticulata khng lm gim vim cp tnh trn m
hnh gy vim chn chut bng carrgeenin (P > 0,05).
* Trn m hnh gy trn dch mng bng
Kt qu cho thy, dch chit t Sargassum swartzii liu 175mg/kg v liu
350mg/kg lm s lng dch r vim gim r rt so vi i chng (P <0,05). Trong khi
Ulva reticulata ch c tc dng liu 350mg/kg (P <0,05). Ngoi ra, dch chit
Sargassum swartzii v Ulva reticulata c 2 liu khng nh hng n hm lng
protein trong dch r vim so vi l chng (P >0,05) v u c tc dng lm gim r rt
s lng bch cu trong dch r vim so vi l i chng (P <0,05). Tc dng gim s
lng bch cu trong dch chit t hai loi to nghin cu c xu hng tt hn khi s
dng Indometacin 25mg/kg.
VII.2.10. Tc dng chng vim mn
Kt qu cho thy, dch chit t Sargassum swartzii liu 175 v 350mg/kg c tc dng
chng vim mn tnh thng qua lm gim trng lng khi u ht so vi chng (P <0,05),
97

tc dng ny c xu hng tt hn khi s dng Prednisolon liu 5mg/kg. Dch chit Ulva
reticulata khng c tc dng chng vim mn (P >0,05).
VII.2.11. Hm lng lipit tng s
- Mu S (Cao Sargassum swartzii):
23,69% trng lng ti
- Mu U (Cao Ulva reticulata):
19,32% trng lng ti
Kt qu phn tch trn cho thy trong hai loi cao th nghim, mu cao rong
Sargassum swartzii cha hm lng lipit tng s cao hn 4,37% so vi mu cao rong
Ulva reticulata.
VII.2.12. Thnh phn axit bo
- Tng cc axit bo no v tng cc acit bo khng no ca hai mu cao rong nghin
cu c s khc bit nhau r rt. mu S, tng cc acit bo no cao gp 2,27 ln so vi
tng cc axit bo khng no (60,50% so vi 26,65%), trong khi mu U th ngc li,
tng cc axit bo khng no li cao gp 1,8 ln tng cc axit bo no (62,70% so vi
34,91%). iu ny cng c ngha l tng cc axit bo no ca mu S cao hn khong
1,73 ln so vi mu U (60,50% so vi 34,91%) cn tng cc axit bo khng no th
ngc li, mu U cao gp khong 2,35 ln so vi mu S (62,70% so vi 26,65%).
- V thnh phn axit bo: Mu U c hm lng axit arachidonic (ARA, C20:4(n6)) cao gp 2,74 ln so vi mu S (1,81% so vi 0, 66%). ARA l mt loi axit bo
thuc h Omega-6, tin cht ca cc Eicosanoid gy vim. V vy, kt qu phn tch ny
c th l mt gi cho vic la chn i tng to bin cho cc th nghim th hot tnh
khng vim.
VII.2.13. Kt qu chit phn lp
T 50 gam cao metanol tng s chit xut t bt to Sargassum swartzii, tin hnh
chit phn lp bng cc dung mi theo th t l n-hexan, diclometan v etyl-axetat. Sau
khi ct ui dung mi thu c 3 loi cao tng ng vi hm lng c th l: cao nhexan chim hm lng cao nht v cao hn rt nhiu so vi hai loi cao cn li
(74,86% so vi 3.80% v 1,60%). Kt qu ny c th cho php d on trong thnh
phn ha hc ca to Sargassum swartzii, hm lng cc cht mu (cc sc t), cc cht
bo v cc cht t phn cc ni chung l tng i ln.
VII.2.14. Hot tnh khng sinh ca 3 loi cao tch t cao metanol tng s ca to S
C 3 loi cao n-hexan, diclometan v etyl-axetat u c th s b hot tnh
khng vi sinh vt nhm xc nh xem thnh phn c hot tnh khng vim c ho tan
trong dung mi no. Kt qu th hot tnh khng sinh cho thy trong 3 mu cao c
th hot tnh khng sinh ch c mu cao diclometan (K hiu C) c hot tnh khng
chng Staphylococcus aureus vi gi tr IC50 l 64g/mL. Gi tr ny cho thy hot tnh
khng sinh ca mu th l tng i tt. iu ny cho php nh hng tip tc phn
tch phn cao ny xc nh bn cht ha hc ca phn dch chit c hot tnh khng
vim. Hai loi cao cn li khng th hin hot tnh khng sinh (hoc c hot tnh nhng
khng ng k).
VII.2.15. Tinh sch cht
a/ Phn cao Diclometan:
Tin hnh SKC phn cao Diclometan tch cc phn on, gp chung cc phn
on c sc k tng t nhau, chng ti thu c hai nhm phn on chnh ng ch
, k hiu l DC1 v DC2 .
- DC1 l nhm p/ cha vt alkaloid d on, c hin di UV bc sng
254 nm, do hm lng tng i thp nn trong th nghim ny chng ti cha kt tinh
v cha lm sch. Qu trnh lm sch s c thc hin trong cc th nghim sau.
98

- Nhm p/ DC2 kt tinh cho tinh th dng bt, mu trng, hm lng khong 30
mg (trong tng s 1,558 gam cao Diclometan em tch, tc l hm lng DC2 chim
gn 2%) . Ra sch tinh th bng cc dung mi thch hp v kim tra sch bng
SKBM. Sc k ch ra trn hnh VIII.3.5.b cho thy vt cht DC2 hin r di UV
bc sng 254nm, c Rf = 0,55 trong h trin khai Chloroform : Metanol (9,5:0,5).
Lm kh hon ton tinh th DC2 v ghi ph xc nh cu trc:
+ Ph khi cho peak M = 123
+ Ph 1H-NMR cho 2 singlet 3,4 v 3,6 l ca hai nhm CH3 ng xa
nhau, 1 singlet 7,8 l ca CH trong vng thm v mt tn hiu 12,7 l ca H lin
kt vi N.
+ Ph 13C cho bit hai tn hiu 28,5 v 30,2 l ca hai nhm CH3 lin kt
vi vng thm. Tn hiu 106 l ca CH bc hai trong vng thm khng b th, cn tn
hiu 140, 149, 151 l ca C bc bn vng thm b th. Tn hiu 156 l ca C
trong nhm acetal CHO. T cc tn hiu ca ph MS, 1H-NMR, 13C-NMR cho thy
phn t c khi lng 123 gm cc nhm sau:
2 CH3
1 CH
3C
1 NH
1 CHO

(30)
(13)
(36)
(15)
(29)

OHC

H3C

Cng thc phn t: C7H9NO

D kin cng thc cu to ca phn t nh sau:

CH3

N1

H
3,5-Dimethyl-4-al-pyrol

b) Phn cao n-Hexan

Bng cc phng php sc k kt hp, chng ti tinh sch c hp cht HI t
cao n-hexan v xc nh c HI cha cht 22-Dehydroclerosterol
(www.lipitbank.jp/cgi-bin/detail.cgi)
+ Danh php quc t: (22E,24S)-24ethylcholesta-5,22,25-trien-3-ol
+ Tn thng dng:
22-dehydroclerosterol
+ Cng thc phn t:
C29H46O1
+ Trng lng phn t trung bnh:
410,675
+ Cng thc cu to:
(Hnh VII.2.15)
+ Nhit nng chy:
acetate 146-150oC
+ D liu ph NMR: acetate 1H-NMR
(CDCl3) 3-CH=4,601 (m), 3-Ac=2,030 (s), 6CH=5,370 (br d), 18-CH3= 0,691 (s), 19-CH3=1,012
(s), 21-CH3=1,011(d), 22-CH=5,243(dd), 23CH=5,171(dd), 24-CH=2,420 (q), 26-CH3=1,646 (d),
27-CH3=4,694 (m), 29-CH3=0,833 (t).
+ D liu ph khi MS: acetate: 452[M+]
(0,3%), 409(0,2%), 392(100%), 377(3%), 363(3%),
Hnh VII.2.15. Cng thc cu to
282(5%), 255(32%), 253(12%), 228(3%), 213(7%).
22-Dehydroclerosterol

99

VII.2.16. Kt qu th hot tnh khng sinh v hot tnh chng oxi ha ca DC2
Sau khi tinh sch v xc nh c bn cht ha hc ca DC2, chng ti tin
hnh phn tch hot tnh khng vi sinh vt v hot tnh chng oxi ha ca cht ny (
dng tinh th).
Kt qu th hot tnh khng sinh v chng xy ha cho thy, thy cht DC2 sau
khi c lm sch li khng th hin hot tnh khng sinh v chng xy ha.
* C th l gii hin tng cht DC2 dng tinh khit khng c hot tnh ca bi
mt s nguyn nhn sau:
- Hot tnh khng sinh ca phn cao CH2Cl2 c th khng phi ch do mt cht
nht nh gy ra m l do tc ng tng tc ca mt s cht cng c mt trong phn
cao ny gy ra. V vy khi tch ring ra, cht DC2 dng tinh sch khng th hin hot
tnh.
- Nguyn nhn th hai c th l do tc ng ca qu trnh tinh sch. Trong qu
trnh lm sch DC2, do nh hng ca dung mi (cha cc cht bo qun...) nn tinh th
b bm cc cht c ngun gc t dung mi, cc cht ny lm nhiu ph ca DC2 nn
chng ti tip tc lm sch loi b cc cht bm ny. C th do qu trnh ra sch
tinh th ko di v tri qua cc iu kin khc nhau (nh nhit , hn hp dung mi...)
nn hot tnh ca cht DC2 gim i ng k. V vy, nng < 128 g/mL, cht
DC2 khng cn th hin hot tnh khng sinh v trn h DPPH, cht DC2 khng th hin
hot tnh chng oxi ha.
- Nguyn nhn th ba c th l do hot tnh khng sinh ca phn cao CH2Cl2 l do
cc cht khc (m chng ti cha tin hnh tinh sch tip) ch khng phi do cht DC2.
Khi , cht DC2 hon ton khng c hot tnh khng sinh.
Nh vy, qu trnh tinh sch cht DC2 c th lm mt i hot tnh khng sinh ca
n. iu ny cn phi kim tra li c th c kt lun cui cng.
VII.3. Kt lun
- thu thp v nh tn c 19 loi rong bin Vit Nam phn b cc tnh min
Bc, Trung v Nam ca Vit Nam.
- xy dng c quy trnh tch dch chit c hot tnh khng vim t cc loi rong
bin Vit Nam bng dung mi metanol; phng php sng lc nhanh cc loi rong to
bin trong khun kh ti c hot tnh khng vim bng dung mi metanol, iu
kin ti u cho vic sng lc cht khng vim t cc loi rong bin Vit Nam l 1 gi
vi nng 0,08 mg/l dch chit khng vim c xc nh.
- Trn c s ca phng php ny, trong tng s 19 loi rong bin c sng lc th
Sargassum swartzii v Ulva reticulata c hot tnh khng vim cao v c tim nng
khai thc c pht hin.
- thu c 300g v 200g dch chit c c c hot tnh khng vim cao t Ulva
reticulata v Sargassum swartzii, tng ng tng loi mt.
- Kt qu th hot tnh khng sinh cho thy: Ulva reticulata c hot tnh khng Bacillus
subtilis vi gi tr IC50 = 62,15 g/ml v Sargassum swartzii c hot tnh khng
Staphylococcus aureus vi gi tr IC50 = 56,53 g/ml.
- tin hnh th hot tnh khng vim ca cc dch chit thu c t Ulva reticulata
v Sargassum swartzii trn ng vt thc nghim (th c tnh cp, tc dng gim au
bng axit axetic, gim au mm nng, gim vim mn bng u ht, gy vim cp mng
bng bng carageenane):

100

+ Dch chit t Sargassum swartzii v Ulva reticulata ung n liu 66g/kg cha
c chut cht. Thuc t c tnh cp, cha tnh c LD50 theo ng ung.
+ Dch chit Sargassum swartzii v Ulva reticulata liu 500mg/kg c tc dng
gim au trn c hai m hnh gy au bng axit axtic v mm nng.
+ Dch chit Sargassum swartzii liu 500mg/kg c tc dng chng vim cp trn
m hnh gy ph chn chut v m hnh trn dch mng bng. Dch chit Ulva
reticulata ch c tc dng chng vim cp trn m hnh gy trn dch mng bng.
+ Dch chit Sargassum swartzii liu 175 v 350mg/kg c tc dng chng vim
mn tnh. Dch chit t Ulva reticulata khng c tc dng chng vim mn tnh.
xc nh c hm lng lipit tng s ca hai mu cao rong Sargassum swartzii v
Ulva reticulata theo th t l 23,69% v 19,32% trng lng ti.
Thnh phn axit bo ca hai mu cao rong Sargassum swartzii v Ulva reticulata
c xc nh. Trong , hm lng axit arachidonic (ARA - C20:4(n-6)) ca cao
Ulva reticulata l 1,81%, cao gp 2,74 ln so vi hm lng axit arachidonic ca mu
cao Sargassum swartzii (0,66%). Tng cc axit bo no v axit bo khng no ca hai
mu cao ny ln lt l 60,50% v 26,65%; 34,91% v 62,70%.
T cao metanol tng s ca Sargassum swartzii sau khi chit phn lp thu c ba
loi cao vi hm lng tng ng nh sau: cao n-hexan (37,43 g) chim 74,86%, cao
diclometan (1,9g) chim 3,80% v cao etyl axetat (0,8 g) chim 1,60%.
Trong ba loi cao n-hexan, diclometan v etyl axetat th ch c cao diclometan c hot
tnh khng Staphylococcus aureus vi gi tr IC50 l 64 g/mL, hai loi cao cn li
khng th hin hot tnh khng sinh (hoc c hot tnh nhng khng ng k).
Phn tch ha hc phn cao diclometan v cao n-hexan cho thy c hai phn cao ny
u cha cht thuc nhm alkaloid. Trong , phn cao diclometan c hai cht quan
tm, k hiu l DC1 v DC2. Cht d on l alkaloid trong phn cao diclometan c
k hiu l DC1, cn DC2 tinh sch c l mt cht kt tinh dng bt mu trng,
hm lng 30 mg. Phn tch cc d liu ph, chng ti xc nh c bn cht ha
hc ca DC2 l 3,5-Dimethyl-4-al-pyrol, c cng thc phn t l C7H9NO.
Phn tch ha hc phn cao n-hexan cho thy trong phn cao ny c 3 cht quan tm,
k hiu l HI, HII v HIII, trong HI xc nh c l 22-dehydroclerosterol, HIII l
cht d on thuc nhm alkaloid (cha xc nh c th) cn HII cha xc nh.
Kt qu th hot tnh khng sinh v hot tnh chng oxi ha cho thy, cht DC2 khng
th hin hot tnh khng cc chng vi sinh vt kim nh nng < 128 g/mL v
khng th hin hot tnh chng oxi ha trn h DPPH (1,1-diphenyl-2picrylhydrazyl).

101

Chng VIII. Nghin cu thm d kh nng sinh cc cht c

hot tnh sinh hc t vi sinh vt bin.
VIII.1. M u
Phn ny trnh by tm tt tm quan trng v li th ca cc nghin cu v ng
dng cc hot cht t vi sinh vt bin. Cc nhm hot cht chnh t vi sinh vt bin
VIII.2. KT QU v tho lun
VIII.2.1 Phn lp v xc nh s lng ca cc nhm vi sinh vt bin trong nc
bin, trm tch bin v trong cc mu rong to, c bin v s huyt Vit Nam
T 81 mu vt bin, chng ti phn lp c 1052 chng VSV, bao gm 552
chng vi khun (chim 52,5% tng s chng), 253 chng nm si (chim 24%), 157
chng x khun (14,9%) v 90 chng nm men (8,6%). Trong cc mu trm tch, s
lng VSV dao ng t 0 n 5,6 . 104 CFU/ml, cao hn so vi s lng VSV trong cc
mu nc bin vi chc ln (0 - 7,6 . 103 CFU/ml). Trong tt c cc mu th s lng vi
khun t cao nht (101 n 104), tip n l nm si, x khun, cn nhm nm men xut
hin t nht (t 0 101 CFU/ml).
Hnh VIII.2.1.a cho thy, s lng VSV trong cc mu trm tch v nc bin
ph thuc vo mn, khong cch xa b, su. S lng VSV trong cc mu trm
tch dao ng t 0 n 5,6.104CFU/g nhiu hn s lng vi sinh vt trong cc mu nc
bin (0-7,6.103CFU/ml). Trong tt c cc mu th s lng vi khun t cao nht (10104), tip n l nm si, x khun, cn nhm mn men xut hin t nht. Trong ,
cc mu nc bin c mn 3% s lng vi khun xut hin t hn vi s lng
102CFU/ml, ring mu nc bin Trng Sa s lng vi khun l 7,6.103CFU/ml cao
hn cc vng bin Ph Yn v Bnh nh. Hu ht mu trm tch bin c mn 2%
Bnh nh v Qung Bnh, s lng vi khun dao ng 104CFU/g, ln hn s lng vi
khunt mu nc bin 10-100 ln (102-103CFU/ml) v cng cao hn 100 ln so vi s
lng vi khun trong cc mu san h v rong trng bin Trng Sa, Nha Trang
(102CFU/ml).
11

T l % s chng VSV khng

VSVK (%)

12

10
10

10

10

8 .5
8

S lng VSV (CFU/ml)

T l % s chng VSV khng VSVK (%)

10

7 .6

5 .5

60
55

10.5

10.6
10.3

10.3

50
40

10

30
9.5

9.3

20
10

10
10

Lng VSV (CFU/g)

12

5.5
0

8.5
<10

80-100

350-500

10001200

Khong cch xa b (m)

0

0
1 - 1 ,5

2 ,5 - 3

Kh? nng K

m n (% )
K h nng K

Slg VSV X1000 (CFU/ml)

S lg V S V x 1 0 0 (C F U / m l)

Hnh VIII.2.1.a. nh hng ca mn v khong cch xa b ln s lng v kh nng

khng VSV kim nh ca vi sinh vt bin Vit Nam
Khc hn so vi cc mu nc v trm tch gn b, cc mu thc vt (rong trng, to)
v ng vt bin (san h, ngao s huyt) vi khun xut hin rt t, cao nht l 102 CFU/g,
cn nm men v x khun hu nh khng c. Theo mt s ti liu, cc chng vi khun
cng sinh vi ng vt bin thng sinh cc cht c hot tnh sinh hc c bit. T c
hi i Ty Dng phn lp c chng Acromonas salamoniada, t c ui vng
102

Yeloowtail Sereola quinqueradiate phn lp c cc chng vi khun Streptococcus

spp. v t tm hm Penaeus monodom phn lp uc vi khun Vibrio hareyi, Vibrio
splendidus. Rut c bin c h vi khun rt phong ph, trong c nhiu chng khng
c vi khun gram (+).
VII.3.2 Sng lc s b, th hot tnh sinh hc ca cc chng VSV bin phn lp
uc (khng sinh, khng nm, gy c t bo, sinh enzim v axit bo khng bo
ho..)
Cc chng VSV bin phn lp c th hot tnh khng khun, khng nm.
Kt qu cho thy, trong s 1052 chng VSV bin c th kh nng i khng th
khng c bt k chng no i khng c vi vi khun kim nh Gram (-) E.coli v
P.aeruginose. C 38 chng vi khun (chim 3,6% s chng), 20 chng x khun
(1,9%), 23 chng nm si (2,1%) v 4 chng nm men (0,38%) khng vi khun Gram
(+) B.subtilis vi ng knh vng v khun dao ng t 7- 46mm, c 1 chng nm men
v 1 chng x khun khng c vi khun gram (+) S.aureus vi hot tnh trung bnh,
ng knh vng v khun t 14-15mm. c bit c 1 chng x khun khng c
nm men gy bnh C.albicans.
Trong 4 nhm VSV bin phn lp c, th nhm x khun c t l s chng c
hot tnh khng khun, khng nm cao nht (12,7%), tip n l nhm nm si (9,1%).
Kh nhiu chng x khun c ng knh vng khng khun ti 40mm, vng khng
nm ti 24mm. Nhm vi khun v nm men hu nh khng c kh nng khng khun,
khng nm, hoc c th hot tnh rt thp. ng knh vng khng khun, khng nm
u di 10mm. c bit, trong qu trnh phn lp tuyn chn tuyn c mt chng
x khun TG1 c kh nng khng chng nm si 55 cng phn lp t Ph Tam Giang,
vng khng 18mm (hnh VIII.2.2).

x khun khng mc ng knh

vng khng 18mm

Hnh VIII.2.2. X khun TG1 khng chng nm si 55 Tam Giang

Theo mt s ti liu, nhiu chng VSV bin c kh nng tng hp mt s c t
gy cht cn trng. Chng ti th hot tnh gy c t bo trn giun t v kh nng
dit su t v su xanh ca 142 chng VSV bin. Kt qu cho thy, c 10% s chng
VSV (8/80) c kh nng dit su hi v u thuc nhm vi khun gram (+) v hu ht
c phn lp t trm tch bin. iu ny cng ph hp vi nhn nh ca mt s tc
gi, rng t trm tch bin su thng phn lp c cc chng vi khun gram (+) c kh
nng sinh c t dit su. Cc chng nm men bin khng c hot tnh dit su v gy
c t bo. Nm si v x khun ch c kh nng gy c t bo, khng c kh nng
103

dit su. Cn nhm vi khun th c c 2 kh nng trn. T l cc chng vi khun c kh

nng dit su cao nht (10%) v gy c t bo ch chim 2,5%. T l chng nm si c
kh nng gy c t bo kh cao, ti 13%, cn x khun ch c 5% tng s chng c
kh nng ny.
Trong s 1052 chng VSV bin c th kh nng i khng vi 5 loi VSV
kim nh, c 3 chng c vng v khun ln nht. 3 chng ny u phn lp c t
trm tch bin. Chng nm si T10.6, chng x khun VN2, v chng vi khun T9.5.
C 3 chng ny c kim tra kh nng gy cht t bo trn i tng l giun t. Kt
qu cho thy, dch nui cy 03 chng x khun VN2, nm si T10.6 v vi khun T9.5
u c hot tnh gy c cho giun t Derodigitata miiler - thc n cho c cnh. Sau 45
pht vi liu lng dch nui cy l 1,5ml c 60-72% s giun t cht. Chng x
khun VN2 c kh nng gy cht giun t cao nht, t 72%. y ch l cc nghin cu
thm d nn tc nhn gy cht giun t chng ti cha c r. Cn nghin cu su hn.
Cc chng x khun, nm si v nm men bin khng c kh nng dit su. Ch
c mt s chng vi khun bin c kh nng ny. Trong s 3 chng vi khun c th
nghim hot tnh dit su t v su xanh th chng vi khun T 9.5 c hot tnh dit su t
tt nht, sau 5 ngy vi lng dch nui cy l 2,0ml c 80% s su cht.
Trong 82 chng c th nghim c trn 50% s chng c hot tnh sinh
amilaza v xenlulaza, ch c khong 12% s chng c hot tnh proteaza. Nhm vi
khun c hot tnh sinh 3 loi enzim cao nht (vng phn gii xenlulaza ti 30mm), sau
n nhm x khun (vng phn gii xenlulaza ti 36mm) v nm si. Cn nhm nm
men c hot tnh enzim thp nht (vng phn gii ch khong 10mm).
Axit bo cha no mch di c quan tm nhiu nht l axit bo loi -3 v -6.
Chng l nhng axit bo cn thit cho sc khe, cho sinh trng v pht trin. Trong
mu nc bin Trng Sa v Nha Trang, chng ti tch c 6 chng nm men bin v
xc nh hm lng axit bo tng s. Kt qu cho thy, hm lng axit bo -3 v -6
ca cc chng ny cn tng i thp. Do , vic tm ra nhng loi nm men c kh
nng sinh ra cc axit bo cn thit -3 v -6 c hm lng cao ng dng lm
nguyn liu thuc v thc n b sung cho nui trng thy sn.
Tt c 6 chng nghin cu, axit bo ch yu trong 2 phn on trung ho v c
cc l Linolic (18:2), axit linolenic (18:3) v axit palmitic(16:0), chim ti hn 80%
axit bo tng s. C 6 chng u khng sinh DHA (22:6) v ch c chng NT2 sinh EPA
(20:5) vi hm lng rt thp 0,13% tng s axit bo. Chng NT1, NT3 v NT8 sinh
axit linoleic (18:2) cao nht, t 28,48-38,55%, cn chng NT2, TS2 v TS1 sinh axit
linolenic (18;3) kh cao, t khong 6,55-8,83% axit bo tng s. Nh vy hm lng
axit bo khng bo ho ca 2 chng NT1 v NT8 vo loi kh. Nh vy, trong 6 chng
th c th tuyn chn 2 chng NT1 v NT8 nghin cu qui trnh sn xut sinh khi
nm men lm thc n cho c.
Hai chng nm men NT5 v NT8 u c kh nng sinh astaxanthin, tuy nhin hm
lng astaxanthin trong sinh khi khng cao (240 v 120 g/g).
Tm li, kt qu sng lc s b hot tnh sinh cht khng khun, khng nm,
sinh axit bo khng no, sc t carotenoit v enzim ngoi bo ca 1052 chng VSV bin
phn lp c, chng ti tuyn chn c 11 chng c hot tnh kh cao, trong c
3 chng x khun khng vi khun Gram(+), khng nm v gy c t bo (VN1, VN2,
VN3), 2 chng nm men sinh axit bo mch di khng bo ho (TS1, NT8), 1 chng
nm men sinh astaxanthin (NT5); 1 chng vi khun Gram (+) sinh c t dit cn trng
(T9.5) v 4 chng nm si khng vi khun Gram (+) (c bit chng T10.6 c hot tnh
rt cao 46 mm).
104

VIII.2.3. Quy trnh tuyn chn, sng lc cc VSV bin sinh cc cht c HTSH
T 1052 chng VSV phn lp t cc mu nc, trm tch ng vt, thc vt bin
chng ti s dng sng lc cc chng c hot tnh sinh cao (ng knh vng
khng >22mm). Cc chng VSV bin c tin hnh sng lc theo cc bc sau :
- Cc chng VSV bin c hot ha trn mi trng thch 1 ngy i vi vi khun, 2
ngy i vi nm men, 7 ngy vi x khun v nm mc.
- Cc chng VSV bin hot ha c cy im trn cc a thch cha cc VSV kim
nh khc nhau gm : B.subtilis, S.aureus, E.coli, P.aeruginosa, C.albicans.
- Nui cy trong t m cc nhit v thi gian khc nhau ty tng c im sinh
trng ca sau chng VSV kim nh. Sau , ly a thch ra quan st, chng vi khun
bin no to vng khng vi VSV kim nh s c gi li tip tc nghin cu (hnh
VIII.2.3.a).
- Cc chng VSV bin c hot tnh c nui cy trn mi trng dch th (MPA i
vi vi khun, Hansen i vi nm men, Gause i vi x khun, Czapek i vi nm
si), trong 1 ngy i vi vi khun, 2 ngy i vi nm men, 7 ngy i vi x khun v
nm si.
- Dch nui cy ca cc VSV bin sau khi loi sinh khi c em th kh nng i
khng vi cc VSV kim nh bng phng php ging thch (mi ging thch nh 200
l dch nui cy VSV).
- cc a thch trong t lnh 4h, sau vo t 370C trong 24h v ly ra c kt
qu.
- Cc chng VSV bin c ng knh vng i khng > 15mm s c s dng tip cho
cc nghin cu tip theo.

cc chng VSV bin to vng khng

cc chng VSV to ng knh vng

khng >15mm

Hnh VIII.2.3.a. Tuyn chn cc chng VSV bin c kh nng khng VSV kim nh
Sau khi tuyn chn, sng lc cc chng VSV bin c kh nng i khng VSV
kim nh, tuyn chn c 3 chng x khun VN1, VN2, VN3 v 1 chng nm si
T10.6 c ng knh vng i khng > 15mm, c bit chng nm si T10.6 c ng
knh vng khng n 46mm. Cc chng x khun VN1, VN2, VN3 c gi sang
Hn Quc nghin cu, cn chng nm si T10.6 c chng ti tin nghin cu
a ra quy trnh nui cy v tch chit cht khng khun.

105

38
13.5
28

12.5

30

16

25

14
12

11.3

10

20 9.2

20

15
10

40

5.5 6
4

2
0

2.5

30
25
20

37

8.4
8.34 8.32

8.3
8.2

20

8.1

15

15

10 8 8

10

7.9

5
0

7.8
5

Nng mui (%)

d-d1(mm)

34
8.3 8.3

35

pH cui

35

35

Vng khng khun (dd1, mm)

40

Sinh khi kh (g/l)

Vng khng khun (dd1, mm)

VIII.2.4. nh hng ca mt s iu kin nui cy ln sinh trng v sinh cht

khng khun ca chng nm si T10.6
i vi qu trnh tng hp cc cht c HTSH c rt nhiu yu t nh hng ti
qu trnh ca chng nh thnh phn mi trng, pH, nhit nui cy, nh sng v
stress. la chn c cc iu kin thch hp nht cho sinh trng v sinh tng hp
cht khng khun ca chng T10.6, chng ti ln lt nghin cu cc yu t trn.
a. nh hng ca thnh phn mi trung nui cy
Thnh phn mi trng nh hng ln n hot tnh khng khun. Cng nh cc
loi VSV khc, nm si cng cn nhng vitamin, cht khong cho sinh trng. Nhu cu
v cht sinh trng ca mt s loi nm c th thay i ty theo iu kin nui cy v
ty theo tui ging. Thnh phn mi trng v kiu nui cy nh hng ln n s sinh
trng v sinh tng hp cht khng khun. Chng T10.6 nui trn mi trng Czapek
cho sinh khi kh v vng khng B.subtilis ln hn so vi mi trng sinh khng sinh,
nhng li khng tt bng mi trng czapek c b sung thm bt ng, bt u tng v
cao nm men. Trn mi trng Cazek ci tin ny, sinh khi tng khong 10%. cn
ng knh vng khng khun tng 25% so vi 2 mi trng cn li. T cc kt qu
ny, chng ti quyt nh chn mi trng Czapek ci tin nui cy v chn kiu
nui cy tnh cho tt c nhng th nghim sau.
b. nh hng ca nng mui trong mi trng
Kt qu th nghim cho thy, nng mui trong mi trng khng nh hng
n sinh trng ca chng nm T10.6. Chng u sinh trng tt mi trng c nng
mui t 0% n 3%. Lp mng nm dy, mu xanh ru thm, ph kn b mt bnh,
dch nui cy c mu vng, trong tt c cc phng n th nghim. Nhng nng
mui li nh hng n kh nng sinh cht khng khun ca chng T10.6 rt r rt. Kt
qu c trnh by trn hnh VIII.2.4.a. nng mui 1% cho vng khng khun ln
nht, nng cao hn, t 2-3% hot tnh gim t 25-50 ln. Khi nng mui l 4% th
n c ch qu trnh tng hp cht khng khun r nht, ng knh vng v khun gim
ti 85%.
c. nh hng ca pH mi trng
pH ban u trong mi trng nh hng r rt ln kh nng khng B.subtilis ca
chng nm si bin T10.6. di pH 5,5-6,5 cho vng khng khun ln nht, t 3437mm, pH qu kim (7,5-8,0) u lm gim vng khng khun ca chng ti 60%. Nh
vy, pH ban u l 5,5-6,5 l thch hp cho sinh tng hp cht khng khun ca chng
nm si T10.6 (hnh VIII.2.4.a). pH cui l 8,0.

5.5

6.5

7.5

pH ca dch nui cy ban u

d-d1(mm)

SKK(g/l)

pH cui

Hnh VIII.2.4.a. nh hng ca pH ban u v nng mui trong mi trng ln kh

nng sinh cht khng khun ca chng nm T10.6
106

d. ng thi sinh trng v sinh tng hp cht khng khun ca chng nm si

T10.6
Nghin cu ng thi sinh trng v tng hp cht khng khun ca chng nm
si T10.6 cho thy, sinh khi kh t cao nht vo ngy th 4 (13,24g/l), cn ng
knh vng khng khun t cao nht vo ngy th 7 (39 2,12). Sau , kh nng khng
khun ca dch nui cy gim dn. Nh vy, t kt qu nhn c cho thy thi gian
dng ln men thch hp nht l sau 7 ngy. Kt qu c trnh by trn hnh VIII.2.4.b.
Sau khi nghin cu nh hng cc iu kin ln s sinh trng v sinh tng hp cht
khng khun ca chng nm si T10.6 chng ti a ra quy trnh nui cy thu dch
khng khun nh sau.
Chng nm si
T10.6 hot ha

Mi trng
Czapek + 1% NaCl

Lc 200vng/ pht
28-300C

4 ngy
Dch ging
cp 1, 2

Mi trng Czapek 1% NaCl, 1%

bt u tng, 1% bt ng
7 ngy

Nui tnh 28-300C

Dch nui cy
chng T10.6

Dch nui cy cha

cht khng khun

Sinh khi chng

nm si T10.6

Hnh VIII.2.4.b. Quy trnh nui cy thu nhn dch khng khun t chng
nm si bin T10.6

Nghin cu tch chit hp cht khng khun

i vi vic tch chit cc cht c HTSH t dch nui cy hoc dch chit, vic
la chn dung mi thch hp l iu quan trng nht. Cc dung mi c s dng phi
p ng cc tiu ch sau: (1) khng nh hng n hot tnh khng khun, (2) c c
tnh bay hi nhanh iu kin nhit thp, (3) phi tch c 2 pha r rt. Trong th
nghim ny, cc dung mi c s dng trong nghin cu l: n-butanol, 2-butanol; nbutyaxetat; etylaxetat, metanol, isopropyl alcohol, clorofom, diclometan.
Vng v khun ca dch nui cy chng T10.6 i vi B.subtilis sau 7 ngy l
37mm. Chng ti s dng dch ny tch cht khng khun vi 6 dung mi: n-butanol,
2-butanol; n-butylaxetat; etylaxetat, cloroform, diclometan. Kt qu cho thy, 6 trong 8
loi dung mi s dng c kh nng tch c 2 pha r rt. Ch c metanol, isopropyl
107

alcohol l khng c kh nng tch pha, nn b loi b. Nhn xt th 2 l, sau khi tch
chit bng 6 dung mi ni trn th cht khng khun th vn gi c hot tnh t 3080% so vi hot tnh ban u. Hot tnh khng B.subtilis khc nhau i vi tng loi
dung mi. Nhng theo tiu ch dung mi phi bay hi nhanh, to 2 pha ring bit, khng
ln dch nui cy, chng ti chn dung mi thch hp l cloroform. Vi dung mi
ny, thi gian tch chit c rt ngn v hot tnh khng khun vn gi c kh cao.
Sau khi cho bay hi clorofom bng my c quay, chng ti thu c dng bt vng mn,
mi hc. Xc nh kh nng khng khun ca khng sinh th cho thy, dung mi
clorofom cho hiu xut tch chit khong 70%, vng khng khun cn 26mm. T cc
kt qu trnh by trn, chng ti a ra qui trnh tch chit cht khng khun t chng
nm si bin T10.6 (hnh VIII.2.4.c).
Dch nui cy chng 10.6
tch sinh khi

Dung mi Clorofom
T l 1:1
Dung dch hn hp

Xc nh hot tnh
khng khun
Nhit
phng

o trn trong
2h
Dung dch hn hp
o trn

Nhit
phng

tnh 3h
Dung dch tch
thnh 2 pha

Nhit
phng
Pha di

Tch pha bng

phu chit
Na2SO4 10g/l

Pha trn

Hn hp clorofom + cht
khng khun
500C,
0,53 at

Dich nui cy sau khi

chit cn li

C quay
600vng/pht
Loi b

Clorofom thu hi

Cht khng khun th

Hnh VIII.2.4.c. Quy trnh tch chit cht khng khun t chng nm si T10.6
VIII.2.5. nh tn khoa hc mt s chng VSV bin c hot tnh sinh hc kh cao
hiu bit su hn v cc chng VSV bin c kh nng sinh mt s cht c
HTSH nh k trn, chng ti tin hnh nh tn khoa hc ca chng. Chng vi khun
T9.5 v chng nm si T10.6 c s b nh tn theo hnh thi, c quan pht sinh bo
t v mt s phn ng sinh ho. Ring 3 chng x khun v 3 chng nm men c nh
tn theo phng php sinh hc phn t.

108

M t c im hnh thi, vi hc ca khun lc chng nm si T10.6

Khun lc trn mi trng Czapek mc nhanh, ng knh 4-5 cm, mu trng, mt
dng xp bng dy 3 mm vng trung tm, vng st mp khun lc c mt dng nhung,
sau 12 ngy c mu xanh lc do to thnh bo t trn. Mt tri khun lc c mu vng
chanh. Khng c git tit. Gi bo t trn nhn pht trin t si nm kh sinh hoc him
hn t si nn, di 40-100 m hoc 150-250 m x 1,5-2,0 m. Thng c 2-3 nhnh
dc thn gi hoc phn ngn gi. Cc nhnh khng to thnh vng r rt. Nhnh c kch
thc 5-20x1,5-2 m. Ngn gi hoc ngn nhnh mang 1 vng th bnh 3-8 chic, th
bnh 6-8x1,5-2 m, phn nh thon dn, hi nhn. Bo t trn hnh cu, gn cu 2,5-3
m, rp nh. Theo bn m t A manual of Penicilia, chng nm si bin T10.6 c tn
khoa hc l Penicillium waksmani Zaleski (hnh VIII.2.5).

Hnh VIII.2.5. C quan pht sinh bo t (a), hnh thi khun lc chng nm si bin (b)

Hai chng nm si TG2, TG3 v chng x khun TG1 phn lp t trm tch ph
Tam Giang c vng v khun khng nm v khng khun kh cao nhng chng ti
cha xc nh c tn khoa hc ca chng.
C 3 chng x khun mc tt trn mi trng c v khng c nc bin. Hnh
thi khun lc kh ging nhau, c mu trng n nu trng, mt khun lc kh, bt. C 3
chng u mc tt sau 6-7 ngy nui cy.
Da trn cy chng loi pht sinh c th nhn thy, chng x khun VN2 c mi
quan h gn gi vi loi Streptomyces malayensis (99,78%). Trnh t nucleotids ca
on 16SrDNA ca chng x khun VN2 cho thy, chng ny c th thuc loi
Streptomyces malayensis. Tuy nhin, khi kim tra hot tnh khng VSV kim nh ca
chng ny th thy chng c hot tnh khng vi khun gram (+) Staphylococcus aureus,
nhng theo ti liu th loi Streptomyces malayensis khng i khng vi
Staphylococcus aureus. Nh vy chng ny cn nghin cu thm v mt phn loi hc,
chng c th l mt loi mi.
Da trn cy chng loi pht sinh, chng x khun VN1 c mi quan h gn gi
vi loi Streptomyces aurantiogriseus NRRL B-5416 (99,15% nucleotit trng lp).
Chng VN3 trn cy chng loi pht sinh c mi quan h gn gi vi
Streptomyces cheonanensis (99,43%), Streptomyces spinoverrucosus (97,12%). S liu
16S rDNA cho thy loi ny c th l Streptomyces sp. m c kh nng khng nm gy
bnh.
Chng nm men bin TS1 phn lp t nc bin o Trng Sa c nghin
cu v hnh thi, nui cy, sinh l, sinh ha, xc nh h thng Ubiquinone, mol
109

%(G+C) v phn tch trnh t nucleotit gen 26S rDNA vng D1/D2. Kt qu ch ra
chng TS1 c c im v hnh thi, nui cy, sinh l, sinh ha ging vi loi Pichia
guilliermondii ngoi tr mt s c im: khng ln men Sucrose, Raffinose; khng
ng ha Galactose, Melibiose, Raffinose, Inulin, D-Ribose, Ribitol, 2-Ketogluconic
axit; ng ha tinh bt tan.
Gen 26S rDNA vng D1/D2 ca chng nm men bin TS1 c 560 nucleotit.
Trnh t nucleotit ny 100% tng ng vi trnh t nucleotit gen 26S rDNA vng
D1/D2 ca loi Pichia guilliermondii c ng k trnh t trong ngn hng gen quc
t. Do c th kt lun chng nm men TS1 thuc loi Pichia guilliermondii.
Chng nm men NT8 (phn lp t nc bin Nha Trang) da theo kt qu phn
loi bng phng php sinh hc phn t, chng thuc loi Candida haemulonii
Cn chng nm men NT5 phn lp t nc bin Nha Trang, hin ang c s
dng sn xut ch phm "nm men Phaffia lm cht mu b sung thc n cho nui
trng thy sn theo cc c im phn loi sinh l sinh ha v da vo trnh t gen 26S
rDNA vng D1/D2 v cy chng loi pht sinh cng cho thy n c quan h gn nht
vi loi P. rhodozyma.
Chng nm si T10.6 phn lp t trm tch bin Qung Bnh thuc loi
Penicillium waksmani Zaleski.
Chng vi khun T9.5 phn lp t trm tch bin c nhiu c im thuc loi
Bacillus sp.
VIII.3. KT LUN
1. phn lp v lm sch c 1052 chng VSV bin t 81 mu nc bin, trm tch
bin v ng thc vt bin Vit Nam c su, mn v khong cch xa b khc
nhau. Trong , c 552 chng vi khun (chim 52,5% tng s chng), 253 chng
nm si (24%), 157 chng x khun (14,9%) v 90 chng nm men (8,6%). T l cc
chng VSV bin phn lp t trm tch bin c kh nng sinh cc cht c HTSH cao
hn so vi nc bin. Cn cc chng phn lp t ng thc vt bin hu nh khng
c hot tnh khng khun.
2. sng lc s b v th kh nng khng 5 chng VSV kim nh (vi khun Gram (-):
E.coli; Pseudomonas aeruginose, vi khun Gram (+): Bacillus subtilis;
Streptococcus..aureus, nm men gy bnh: Candida albicans) ca 1052 chng VSV
bin. Ph khng khun ca cc chng VSV th nghim rt hp. Khng c chng no
khng vi khun Gram (-), c 8,1% s chng c hot tnh khng vi khun gram (+) v
khng nm, nhng ch c 1,2% tng s chng c hot tnh cao (vng v khun t 2239 mm). Trong 4 nhm VSV bin phn lp c, th nhm x khun c t l s chng
c kh nng khng khun, khng nm cao nht (6,8%), tip n l nhm nm si
(9,1%). Nhm vi khun v nm men hu nh khng c kh nng khng khun, khng
nm, hoc c th rt thp.
3. th kh nng dit su hi, gy c t bo ca 142 chng VSV bin. C 5,7% s
chng c kh nng dit su t (Plutella xylostella) v su xanh (Heliothis armigera).
C 5,1% s chng c kh nng gy cht cho giun nhiu t Derodigitata Miiler .
4. xc nh kh nng sinh cc enzim ngoi bo amylaza, xenlulaza v proteaza ca
82 chng VSV bin. Trn 50% s chng c hot tnh sinh amilaza v xenlulaza, ch
c khong 12% s chng c hot tnh proteaza. Cc chng c hot tnh enzim ngoi
bo phn ln u c phn lp t cc mu nc v trm tch ven b.
5. xc nh hm lng axit bo khng bo ho mch di ca 6 chng nm men bin,
hm lng. C 6 chng u khng sinh DHA (22:6) v ch c chng NT2 sinh EPA
(20:5) vi hm lng rt thp 0,13% tng s axit bo. Chng NT1, NT3 v NT8 sinh
110

Axit linoleic (18:2) cao nht, t 28,48-38,55%, cn chng NT2, TS2 v TS1 sinh Axit
linolenic (18;3) kh cao, t khong 6,55-8,83% axit bo tng s.
6. tuyn chn c 11 chng VSV bin c hot tnh sinh hc cao: 3 chng x khun
khng vi khun Gram (+), khng nm v gy c t bo (VN1, VN2, VN3), 3 chng
nm men sinh axit bo mch di khng bo ho v sinh astaxanthin (TS1, NT8,
NT5), 1 chng vi khun Gram (+) sinh c t dit cn trng (T9.5) v 4 chng nm
si khng vi khun Gram (+) (trong T10.6 c vng v khun rt ln >45mm).
7. a ra qui trnh nui cy v qui trnh tch chit cht khng khun t dch nui cy
chng nm si T10.6.
8. tin hnh nh tn mt s chng VSV bin c HTSH cao. C 3 chng x khun
u thuc ging Streptomyces.. Chng VN1 c ti 99,15% nucleotids trng lp vi
cc nucleotids ca loi Streptomyces aurantiogriseus NRRL B-5416 ; Cn chng
VN2 c mi quan h gn gi vi loi Streptomyces malayensis (99,78%), Cn chng
VN3 trn cy chng loi pht sinh c mi quan h gn gi vi Streptomyces
cheonanensis (99,43%). Chng nm men TS1 thuc loi Pichia guilliermondii..
Chng NT8 thuc loi Candida haemulonii. Chng NT5 thuc loi Phaffia
rhodozyma. Chng nm si T10.6 thuc chi Penicillium waksman Zaleski. v chng
vi khun T9.5 thuc chi Bacillus sp.

D. Cc nghin cu ng dng sn xut cc sn phm t

dc liu bin
Chng IX. NGhin cu cng ngh sn sut thc n chc
nng b sung omega 3 phng v h tr iu tr tim mch
IX.1. m u
Phn ny c chia thnh 4 mc ln nh sau:
IX.1.1. Tnh hnh ngi mc bnh tim mch, ri lon chuyn ho lipoprotein
cholesterol cao
Mc ny trnh by tm tt v mi nguy him ca bnh tim mch v a ra cc s
liu thng k v t l mc mt s bnh (c bit l bnh tim mch) v t l t vong trn
th gii v nc ta.
Trn con s t vong ton th gii, t vong do bnh tim mch ng hng u
(23%), trn c tt c cc bnh ung th cng li (19%), cc bnh h hp (18%) v cc
bnh nhim trng v k sinh trng (14%). Trong cc thp k qua, nhiu nc nh Thy
in, Phn Lan, c, Hoa K... thc hin nhiu bin php phng nga bnh mch
vnh v h t c mt s kt qu kh quan. Vit Nam, cng vi s bng n ca
th ho, s pht trin khoa hc v cng ngh... lm thay i li sng ca ngi
dn: tnh ti t vn ng, cng thng thn kinh - tm l v ch n ung tha calo.
iu ny dn n tng nhanh s lng ngi b tha cn, bo ph, bnh nhn mc cc
bnh tim mch, h hp, bnh nhn ung th. T l ngi mc bnh tim mch v t l cht
do cc bnh ny ang chim v tr s mt.
Nhn chung, mt s nc pht trin, s t vong tim mch trc kia rt cao, gn
y ang gim mt cch r rt. Tri li, cc nc ang pht trin, t vong tim mch
hin nay khng ng hng u, nhng li ngy cng c xu hng gia tng. l nhng
nhn xt lm chng ta phi suy ngh. S tng ca bnh tim mch v t l cht do bnh
tim mch ngy cng cao khin cho mi ngi bng hong v quan tm n ch n
phng bnh tim mch hn. Nhiu loi thc phm b sung omega 3, DHA v cc cht
dinh dng c tc dng ngn nga bnh tim mch xut hin v ngy cng nhiu.

111

IX.1.2. Ch n trong phng v iu tr ri lon chuyn ho lipoprotein mu.

Theo s hiu bit hin nay ba yu t nguy c c xc nh, l ht thuc l,
tng huyt p v hm lng cholesterol trong mu cao. Trong , hm lng cholesterol
l nguyn nhn chnh gy ra cc bnh v tim mch. iu tr h cholesterol mu, iu
chnh c cc ri lon v lipoprotein mu khng nhng c kh nng ngn chn m cn
phc hi c nhng tn thng va x thnh mch vnh.
Mc ny trnh by tng quan v lipit v lipoprotein mu, vai tr ca cc cht dinh
dng (gluxit, cht x, protein, axit bo no v khng no...) trong gim ri lon chuyn
ha lipoprotein mu, v nguyn tc xy dng ch n trong ri lon chuyn ho
lipoprotein.
Mt iu ng quan tm l: trong nhng nm gn y c nhiu cng trnh
nghin cu v chng minh v vai tr tch cc ca cc loi axit bo omega - 3 i vi
phng chng cc bnh tim mch. Cc loi c, du c cha nhiu axit bo ny nh cc
Eicosapentaenoic (EPA) v Docosahexaenoic (DHA). Cc loi thc vt ni bin v
sng h tng hp cc axit bo ny v chng l ngun thc n cho c, hi sn v ng vt
c v bin. Trong nhng nm 50 ngi ta bit vai tr gim cholesterol ca axit bo
cha no n-6-linoleic c nhiu trong thc vt v by gi ngi ta cng nhn thy du c
cng c tc dng gim cholesterol tng t. Cc quan st trn thc a cho thy cc b
tc Eskimo t l mc mch vnh thp hn so vi ngi an Mch nh ch n ca h
nhiu du c bin hn. Cc nghin cu cho thy cc axit bo omega - 3 khng nhng
gim cholesterol m cn gim c triglycerit nhng ngi c triglycerit cao. Cc axit
bo omega - 3 cn c tc dng tt phng chng lon nhp tim, rung tm tht, huyt
khi v iu chnh phn no huyt p trong tng huyt p th nh. Cc axit bo omega-3
ngun gc thc vt (axit - linolenic - ALA) cng c tc dng tt i vi bnh tim
mch. ch n giu ALA, nguy c t vong do bnh tim mch vnh gim ti 50%.
T cc quan st trn ngi ta cho rng, ch n hng ngy cn tng cc axit bo n - 3
phng cc bnh tim mch c th l mi tun nn c 2-3 ln n c, thay th c cho
tht. Tt c cc loi c v hi sn u cha cc axit bo n - 3, ngay c khi lng lipit thp
trong mt s hi sn. i vi nhng ngi khng thch n c v hi sn c th s dng
du c mi ngy 2 - 3 g. B sung du c c tc dng lm gim c ngha triglycerit
mu trn c nhng i tng c triglycerit bnh thng v triglycerit cao ( 2 mmol/l).
Vic b sung khong 9 - 13 g du c/ngy (tng ng vi 1,7 - 7 g axit bo omega 3/ngy) th s lm gim 20 - 25% triglycerit ngi c lng triglycerit bnh thng v
gim 26 - 33% triglycerit ngi c triglycerit tng.
Thm vo , cc nghin cu v ch phm Omega-3 c thnh phn chnh l cc
axit bo khng no omega-3 trn ng vt thc nghim trong khun kh ti KC09.15
(2003-2005) cho thy: Khi dng Omega-3 trong thi gian 30 ngy khng gy nh hng
g n s pht trin bnh thng v th trng v trng lng cc tng gan, lch, thn ca
chut nht trng thc nghim. Nghin cu nh hng ca Omega-3 trn s tiu th thc
n (TA), nc ung v cht thi ca chut nht trng cho thy: S tiu th thc n, nc
ung gia cc nhm chut th nghim khng c s khc bit, khi so snh cng thi
im v lng cht thi trung bnh gia cc nhm nghin cu khng c s khc bit, khi
so snh cng thi im. Nghin cu nh hng ca Omega-3 trn chc phn to mu
v chc nng gan, thn cho thy: vi mc liu Omega-3 dng (0,25 g/kg v 0,30 g/kg
th trng) ung 30 ngy, trn ng vt thc nghim khng thy tn thng gan, thn,
lch. Hnh thi gan, thn, lch ca nhm chng so vi nhm thuc khng c s khc bit g
ng k. Thuc khng c vi gan, lch, thn v gc m bnh hc. Hn na, ch
phm Omega-3 c tc dng hn ch s suy gim nng lc tm thn kinh trn chut nht
trng thc nghim.
112

Nh vy b sung du c s l mt gii php an ton, c tc dng iu tr tt cho nhng

ngi b triglycerit mu cao v gp phn ci thin tr nh.

IX.1.3. Mt s nguyn liu c li cho phng nga bnh tim mch.

Mc ny gii thiu mt s nguyn liu c li cho phng nga bnh tim mch nh:
Go lt c tc dng gim thiu nguy c mc bnh nhi mu c tim, kim sot huyt p,
lm gim choletsterol, lm gim nguy c ri lon chuyn ha. Vng gp phn h tr sc
khe tim mch, h hp v lm gim cholesterol. u tng cng l thc phm c gi tr
dinh dng cao, vi thnh phn isoflavon c tc dng lm gim nguy c mc bnh tim
mch, h cholesterol v c bit tt cho ph n thi k mn kinh.
IX.1.4. Cng ngh p n cao p.
Phn ny gii thiu v cng ngh p n cao p, chc nng ca mt my p n
cao p v mt s li ch ca my p n trong sn xut thc phm.
IX.2. Kt qu v tho lun
IX.2.1. Nghin cu qui trnh cng ngh sn xut thc n chc nng phng v h
tr iu tr bnh tim mch qui m cng nghip.
IX.2.1.1. Phn tch thnh phn mt s nguyn liu chnh
Trn c s nhu cu dinh dng ca ngi b bnh tim mch v tham kho cc ti
liu, chng ti la chn mt s loi nguyn liu chnh cho sn xut thc n chc
nng phng v h tr iu tr bnh tim mch, l go, go lt, u tng, vng. Phn
tch thnh phn dinh dng ca cc loi nguyn liu trn, kt qu c trnh by bng
IX.2.1.a v bng IX.2.1.1.b
Bng IX.2.1.1.a. Thnh phn c bn ca mt s nguyn liu s dng cho sn xut thc
n chc nng phng v h tr iu tr bnh tim mch
Thnh phn

Go

Go lt

u tng

Vng

Tinh bt (%)

76,2

75,2

24,6

16,6

Protein (%)

7,3

7,8

34,0

18,1

Lipit (%)

1,4

1,3

18,4

57,6

m (%)

14,2

13,2

14,0

7,6

Cht x (%)

0,6

2,9

4,5

3,4

Bng IX.2.1.1.b. Kt qu phn tch hm lng lipit v axt bo ca vng trng ( xt v)

TT

Thnh phn

n v tnh

Hm lng

% cht kh

57,61

Lipit

Axt Palmitic

% lipit

9,48

Axt Stearic

% lipit

6,46

Axt Oleic

% lipit

38,35

Axt Linoleic (omega 6)

% lipit

43,35

Axt Linolenic (omega 3)

% lipit

2,36

Kt qu phn tch cho thy go rt giu tinh bt (76,2%), trong khi u tng
l nguyn liu c hm lng protein rt cao (34%) v vng c hm lng cc axt bo
113

khng no rt phong ph. Cc axt bo quan trng nh axt oleic (axt bo khng no mt
ni i MUFA) trong vng chim n 38,35% hm lng lipit. Cc axt bo omega 6
v omega 3 cng chim n 43,35 v 2,36 % lipit tng ng. Bn cnh , go lt l loi
nguyn liu rt giu cht x, c bit lng du trong lp v cm ca go lt c tc
dng lm gim cholesterol LDL C (Low Density Lipoprotein - Cholesterol) rt tt.
y l cc loi ng cc sn c Vit Nam, khi kt hp li s to ra mt ngun thc
phm cn bng v dinh dng, rt tt cho phng v iu tr bnh tim mch.
IX.2.1.2. Nghin cu ti u ha cc iu kin cng ngh ca qu trnh p n hn
hp nguyn liu go, go lt, u tng, vng bng ma trn ton hc thc nghim
Doehlert.

Hnh IX.2.1. B mt p ng ca hm mc tiu Y1 - ho tan ca bng (%)

.

Hnh IX.2.2. B mt p ng ca hm mc tiu Y2 - n ca bng (ln)

114

Nh trnh by phn tng quan, cng ngh p n cao p l mt cng ngh

tin tin, hin nay ang c s dng rng ri cc nc trn th gii v Vit Nam
trong sn xut thc phm nh cc loi ng cc n lin, snack. Vi cc nguyn liu c
hm lng tinh bt cao nh go hoc ng th qu trnh p n din ra kh d dng,
nhng vi cc loi nguyn liu l u tng, vng c hm lng protein, lipit cao my
rt d b tc, sn phm thng sng hoc chy. to cho sn phm c c hm lng
cc cht dinh dng cn bng, c bit l s dng c ngun cht x t go lt, th
vic la chn cc iu kin cho qu trnh p n l rt cn thit sn phm to ra c
chn, n, ho tan tt, ph hp vi yu cu ca mt sn phm n lin.
Ma trn Doehlert c thit lp trn c s ca cc bin s l m ca
nguyn liu hn hp (go t + go lt + u tng + vng = 40% + 40% + 10% + 10%)
trc khi a vo p n (X1), bin i t 14 - 22% v tc cp liu cho my p n
(X2), bin i t 575 - 675 g/pht ti u ha cc hm mc tiu l ho tan (Y1) v
n ca bng (Y2). Phn tch chi tit cc kt qu thu c, kt hp b mt p ng ca
hm mc tiu Y1 v Y2, c th kt lun rng khi m ca hn hp nguyn liu ban
u a vo p n t 18,0 - 19,4% v tc cp liu t 623 - 640 g/p (tng ng
37,38 - 38,40 kg/h) th bng c ho tan v n ln nht, tng ng l 37,1% v 6,8
ln.
IX.2.1.3. Nghin cu cng thc phi trn to sn phm thc phm chc nng phng
v h tr iu tr bnh tim mch.
Sau khi p n, hn hp go + go lt + u tng + vng c nghin mn v
phi trn vi cc loi bt tht, malto - dextrin v sau b sung ch phm omega 3 t c
bin to ra thc phm chc nng phng nga v h tr iu tr bnh tim mch. Mc
tiu ca qu trnh phi trn l to ra sn phm c t l cc thnh phn protein, lipit,
gluxit ph hp vi nhu cu dinh dng ca bnh nhn tim mch, c bit sn phm phi
giu hm lng axt bo khng no omega 3 ci thin chc nng chuyn ho lipit mu,
huyt p bnh nhn tim mch. ng thi sn phm phi c gi tr cm quan tt. Sau
khi lm cc th nghim s b, mt s cng thc phi trn c tin hnh v trnh by
bng IX.2.1.3.a. Thnh phn dinh dng ca cc cng thc phi trn c trnh by
bng IX.2.1.3.b.
Bng IX.2.1.3.a. Mt s cng thc phi trn to sn phm thc phm chc nng phng
v h tr iu tr bnh tim mch
Cng thc
Nguyn liu
1
2
3
4
Hn hp Go + go lt +
85
80
80
75
u tng + vng (%)
Bt tht (%)
5
10
5
10
Malto dextrin (%)
5
5
10
10
Ch phm omega 3 (%)
2
2
2
2
Ph gia (%)
3
3
3
3
Kt qu bng IX.2.1.3.b cho thy xt v thnh phn ho hc th c 4 cng thc
u p ng c nhu cu dinh dng cho ngi bnh tim mch. l: nng lng t
gluxit > 70%, nng lng t protein: 14-15%, nng lng t lipit <15%. Tuy nhin, v
tnh cht l hc, cng thc 4 c nht thp nht (558 Cp) v ho tan cao nht
(44,5%). y l nhng ch s rt tt nh gi cht lng mt sn phm ng cc n
lin do nht thp v ho tan cao lm tng kh nng hp thu ca sn phm, mt
khc lm gim cm gic chng ngn khi n. Hn na, im cm quan ca cng thc 4
115

cng t c cao nht (18,56 im). Do vy, cng thc 4 c la chn cho sn
xut thc phm chc nng phng nga v h tr iu tr bnh tim mch.
Bng IX.2.1.3.b. Thnh phn ho l ca mt s cng thc phi trn to sn phm thc
phm chc nng phng v h tr iu tr bnh tim mch
n v
Cng thc
Thnh phn
tnh
1
2
3
4
Tnh cht l hc
nht 500C
Cp
386
391
463
558
ho tan
%
36,5
37,1
39,3
44,5
Thnh phn ho hc
Protein
%
12,78
15,21
12,40
14,48
Lipit
%
4,02
4,16
3,91
3,87
Gluxit
%
71,28
69,04
72,60
70,12
m
%
4,6
4,7
4,9
5,0
Omega 3
mg/100 g
680,58
675,13
675,13
670,24
im cm quan
im
16,59
17,20
17,93
18,56
Tuy nhin, vi t l phi trn ch phm omega 3 l 2%, sn phm c nh gi
l cha ni bt c hng v thm ngon do b nh hng bi mi tanh ca ch phm
omega 3. Do vy, t l ch phm omega 3 c iu chnh xung thp hn sao cho va
m bo c cht lng ca sn phm, va tng c gi tr cm quan. Kt qu cho
thy vi t l ch phm omega 3 s dng t 1,0-1,5% th sn phm thc n chc nng
cho ngi tim mch vn gi c hng v c trng. Tuy nhin, khi tng t l ch
phm omega 3 ln 2,0% th sn phm c mi hi tanh do ch phm omega 3 c chit
xut t c bin v iu ny nh hng rt ln n gi tr cm quan ca sn phm. Do
vy, chng ti la chn t l b sung ch phm omega 3 l 1,5% sn phm va c
gi tr cm quan tt, va p ng c nhu cu v omega 3 cho c th trong 1 ngy.
Theo tiu chun ca T chc Y t Th gii th nhu cu omega 3 ca con ngi l 500
mg/ngy (Kris Etherton, P. M., Harris, W. S., Appel, L. J. (2002). Fish consumption, fish oil, omega
3 fatty acids, and cardiovascular disease. American Heart Association 106, p. 2747 2757).
IX.2.1.4. Qui trnh cng ngh sn xut thc phm chc nng phng v h tr iu tr
bnh tim mch
Chun b nguyn liu:
- u tng c tch v trc khi trn vi cc nguyn liu khc bng my xay xt.
- Go trng: lm sch, loi b sn, tru.
- Go lt: ngm nc nhit thng trong thi gian 60 pht. Sau , vt ra
ro v trn vi cc loi nguyn liu khc.
p n: Qu trnh p n c thc hin trn my p n mt trc
- Hn hp go trng: go lt : u tng: vng c phi trn vi t l 40% : 40% :
10% : 10% c p n iu kin m ca hn hp nguyn liu l 18-19%, tc
cp liu 38-40 kg/h.
Nghin mn
Bng go + u tng + vng c em nghin mn trn my nghin ba thu c
bt hn hp.
Phi trn v ng ti
- Cc loi bn thnh phm c phi trn bng my trn theo t l: Bt G+T+V:
bt tht: malto dextrin: ch phm omga 3: ph gia = 75% : 10% : 10% : 1,5% : 3,5%.
116

- Thng s ca my trn: Tc vng quay bng 120 vng /pht. Thi gian 10
pht/m 20 kg.
- Sn phm c cn nh lng 300g/ti v ng trong mng PE/Al/PP c tc dng
chng m, chng nh sng. Sau c ng hp giy. Sn phm sau khi c kim tra
cc ch tiu cht lng chnh mi c nhp kho. Quy trnh sn xut c trnh by trn
s di y:
u tng

Sy
600C/3h

Tch v
Vng

u tng
mnh

Go
+ go
lt

Rang chn

Phi trn

p n cao p

Nghin

Hn hp bt go +
go lt + u tng
+ vng

Phi trn
Bt tht, malto-dextrin

ng gi

Thnh phm
117

Ch phm
Omega 3

IX.2.2. Nghin cu xc nh ch tiu l ho, vi sinh v th nghim lm sng sn

phm thc n chc nng phng v iu tr bnh tim mch ti Bnh vin.
IX.2.2.1. Xc nh thnh phn dinh dng, ch tiu ho l, vi sinh, ca sn phm
Sau khi xc nh c cc iu kin cng ngh ca qui trnh sn xut thc n
chc nng phng v h tr iu tr bnh tim mch, vic sn xut c tin hnh. Sn
phm to ra c phn tch cc ch tiu cht lng, kt qu c trnh by cc bng
IX.2.2.1.a-c.
Cc ch tiu cm quan:
- Trng thi: dng bt mn
- Mu sc:
vng sng
- V:
ngt, m
- Hng:
thm c trng
Bng IX.2.2.1.a. Cc ch tiu cht lng ch yu, gi tr dinh dng
STT

Tn ch tiu

n v tnh

Hm lng

%
Cp
L
a
b

41,5
542
81,36
1,21
13,74

%
%
%
%
mg/100g
mg/100g
mg/100g
%
%

4,80
71,08
15,42
3,44
457,17
39,6
2846,61
1,85
0,15

Tnh cht l hc
1
2
3

ho tan
nht 500C
mu

Thnh phn ho hc
4
5
6
7
8
9
10
11
12

m
Hm lng Gluxit
Hm lng Protein
Hm lng Lipt
Hm lng omega 3 (linolenic)
Hm lng DHA
Hm lng axit linoleic
Hm lng tro tng s
Hm lng tro khng tan trong HCl 10%

Bng IX.2.2.1.b. Cc ch tiu vi sinh vt v cc c t vi nm ca sn phm

STT
1
2
3
4
5
6
7
8
9

Tn ch tiu
E.coli
Coliforms
S. aureus
Cl.perfringens
Salmonella
B. cereus
TSBTNM- M
TSVSVHK
Aflatoxin
118

n v tnh

Hm lng

kl/g
kl/g
kl/g
kl/g
kl/25g
kl/g
kl/g
kl/g
g/kg

0
0
0
0
0
0
0
2 x 102
Khng pht hin

Bng IX.2.2.1.c. Hm lng kim loi nng trong sn phm

STT
2
3
4
5
6
7

Tn ch tiu
Ch (Pb)
ng (Cu)
Asen (As)
Thu ngn (Hg)
Km (Zn)
Cadimi (Cd)

n v tnh

Hm lng

mg/kg
mg/kg
mg/kg
mg/kg
mg/kg
mg/kg

0,02
< 0,01
< 0,002
< 0,002
8,02
< 0,01

Sn phm c xy dng tiu chun c s v c B Y t cp Chng nhn

tiu chun sn phm s: 6585/2007/YT CNTC (hnh IX.2.2.1.a.) vi tn gi Thc
phm chc nng: Cefish Ci thin chc nng chuyn ho lipt mu, huyt p bnh
nhn tim mch. Sn phm c bao gi trong mng PET/Al/PE, v bn ngoi l hp
Douplex. Khi lng tnh 300 g/hp (hnh IX.2.2.1.b).

Hnh IX.2.2.1.a. Giy chng nhn tiu chun sn phm CEFISH

119

Hnh IX.2.2.1.b. Nhn hp thc phm chc nng phng nga

v h tr iu tr bnh tim mch
IX.2.2.2. Nghin cu th nghim lm sng ti Bnh vin
nh gi chnh xc hiu qu ca vic s dng thc n chc nng n kh nng
lm gim ri lon chuyn ho lipit mu, bnh n huyt p, chng ti th nghim lm
sng trn 24 bnh nhn tim mch s dng thc n chc nng trong vng 2 thng v ng
thi so snh vi 24 bnh nhn i chng (b bnh tim mch nhng khng s dng thc
n chc nng). Sau mi thng, kim tra cc ch s lipit mu, cc chc nng gan, thn,
huyt p, nhp tim v cc du hiu c nng ca tng bnh nhn trong 2 nhm trn. Kt
qu thu cho thy:
- V chc nng chuyn ho lipit mu
Bnh nhn nhm th nghim sau 2 thng c cholesterol gim t 5,5 mmol/L
(vt tr s bnh thng) xung cn 5,1 mmol/L (nm trong tr s bnh thng), c
ngha thng k (P<0,001). nhm i chng th cng sau thi gian 2 thng ch s
cholesterol khng c ci thin m cn tng ln n 5,8 mmol/L (P <0,001).
i vi HDL C (High Density Lipoprotein - Cholesterol), l cholesterol c li
th sau th nghim, tr s ny nhm th nghim tng t 1,2 mmol/L ln 1,6 mmol/L,
trong khi nhm i chng gi tr ny khng i. i vi LDL- C (Low Density
Lipoprotein - Cholesterol), l cholesterol xu th sau thi gian th nghim, gi tr ny
gim i t 3,3 mmol/L n 2,5 mmol/L nhm th nghim, trong khi nhm i
chng gi tr ny khng c ci thin (vn gi l 3,4 mmol/L nh trc khi th
nghim). i vi ch s triglyxerit, c 2 nhm i chng v th nghim, sau thi gian
2 thng th c s bin i khng ng k (P>0,05).
nhm th nghim:
- S bnh nhn c hm lng cholesterol trong mu cao (vt tr s bnh thng) trc
khi th nghim l 14 ngi, sau khi th nghim ch cn li 8 ngi.
- S bnh nhn c hm lng HDL C tt (> 0,9 mmol/L) trc khi th nghim l 21
ngi sau th nghim tng ln 24 ngi.
- S bnh nhn c hm lng LDC C vt qu mc bnh thng trc th nghim l
11 ngi nhng sau th nghim gim xung khng cn ngi no.
120

 nhm i chng: kt qu chuyn bin v chc nng chuyn ho lipit mu l

khng ng k. Sau 2 thng th nghim, s bnh nhn c ch s cholesterol xu khng
gim trong khi , s ngi c ch s cholesterol tt khng tng.
- V huyt p v nhp tim
nhm th nghim: Huyt p tm thu v huyt p tm trng c xu hng chuyn
v gi tr bnh thng, c ngha l chuyn bin theo hng c li.
nhm i chng: huyt p tm thu v huyt p tm trng vn duy tr gi tr
tng ng nh trc khi th nghim c ngha l huyt p khng c ci thin.
V nhp tim, c 2 nhm i chng v th nghim u khng c g bin i.
- V cc chc nng gan, thn
c hai nhm i chng v th nghim th cc chc nng gan, thn u khng c
g thay i bt thng trong th nghim. Ngoi ra, cc bnh nhn n thc n chc nng
hon ton khng c du hiu no b au bng, a lng, kh tiu, bun nn.
Nh vy thc n chc nng cho ngi tim mch c li cho vic ci thin chc nng
chuyn ho lipit mu, huyt p bnh nhn tim mch.
IX.2.3. Sn xut th nghim thc phm chc nng Cefish
Sn phm Thc phm chc nng: Cefish Ci thin chc nng chuyn ho lipt
mu, huyt p bnh nhn tim mch c sn xut trn dy chuyn thit b ti
Xng thc nghim Vin Cng nghip Thc phm vi khi lng 350 kg, trong c
120 kg c mang i th nghim trn bnh nhn tim mch ti Bnh vin a khoa tnh
H Ty. Phn cn li gii thiu v phn phi ti Vin Cng nghip Thc phm, Cng ty
Dc phm Vit Hng.
IX.3. Kt lun
1. xy dng c quy trnh cng ngh sn xut thc phm chc nng phng nga v
h tr iu tr bnh tim mch. Cc thng s ca qui trnh cng ngh c ti u v
iu kin sn xut qui m cng nghip.
2. xc nh c cc ch tiu l, ho, vi sinh ca thc phm chc nng phng nga
v h tr iu tr bnh tim mch. Sn phm c thnh phn dinh dng cn bng, giu
omga 3 ph hp vi nhu cu ca ngi bnh tim mch.
3. th nghim lm sng sn phm trn 48 bnh nhn tim mch iu tr ti Bnh vin
a khoa tnh H Ty cho kt qu tt. Sn phm c chng nhn tiu chun cht
lng ti Cc V sinh an ton thc phm B Y t s: 6585/2007/YT CNTC vi
tn gi Thc phm chc nng: Cefish Ci thin chc nng chuyn ho lipt mu,
huyt p bnh nhn tim mch.
4. sn xut c 350 kg sn phm Cefish trong 120 kg sn phm phc v cho th
nghim lm sng, 230 v ang c gii thiu cc knh phn phi thc phm
chc nng v bc u tip cn th trng.

121

chng X. Nghin cu qui trnh sn xut thc phm chc

nng tng cng th lc cho vn ng vin t Sao bin
X.1 m u
Phn ny c chia thnh cc mc sau:
X.1.1. Vn ng th thao vi trao i cht v chuyn ho nng lng trong c th.
Mc ny nu ra cc yu t c vai tr quan trng nh hng n nng lc hot
ng th lc cng nh kh nng chu ng ca vn ng vin.
X.1.2. Hot cht thin nhin trong thc phm l ngun dinh dng v l thc
phm Thuc c gi tr v rt quan trng vi sc kho con ngi
Mc ny trnh by vai tr v tc dng ca cc protein, axit amin, vitamin v cc
nguyn t vi lng c b sung qua ng thc phm b sung dinh dng.
X.1.3. Testosteron v cc hormone steroid vi s mt mi trong luyn tp th thao
v hun luyn qu sc v tc dng ca chng trong c th khi c b sung qua
ng thc phm chc nng.
Mc ny trnh by vn tt v bn cht ha hc, vai tr v tc dng ca mt s
hoocmon steroit nh: testosteon, progesteron, estradiol, cortisol. c bit l testosteron
khi c chuyn thnh dihydrotestosteron c chc nng bit ho sinh hc trong qu
trnh pht trin phi, m bo s trng thnh, dn ti dy th v bo tn c tnh sinh
dc cng nh bn nng sinh dc ca n ng (ging c), m bo sinh sn ngi
nh s to tinh trng, c vai tr kch thch to hng cu, tng hp Hemoglobin trong
nguyn hng cu. Testosteron trong c th tng hay gim u gy ra s thay i cc
cng sut ca cc h thng trao i cht v chuyn ho nng lng. T thc tin c
cc nh khoa hc chng minh khi c th trng thi xung sc th hm lng
testosteron lc ny t c n nh cao v ngc li khi c th trng thi mt mi
th hm lng testosteron trong mu lc ny b tiu gim. vn ng vin th thao
hm lng testosteron trong mu gim di 0,69 nmol/l i vi n hay di 3,47 nmol/l
i vi nam th c coi l trng thi in hnh luyn tp qu sc v lc ny cn c
tng cng b sung ti to hi phc
X.1.4. Mi lin quan gia Testosteron, Lactat dehydrogenaza (LDH) v axit lactic
trong vn mt mi v hi phc ca vn ng vin.
Mt vn rt quan trng trong dy chuyn phn ng sinh ho trong c th l khi
hm lng testosteron trong mu tng th t l thun vi hot enzym LDH tng km
theo s gim xung ca axit lactic. Enzym LDH tham gia vo qu trnh gii phng axit
lactic trong c, m nh ta bit axit lactic chnh l nguyn nhn gy ra mt mi ca c
th. Trong th thao axit lactic l ch tiu c trng v c tin cy cao trong vic theo
di v khng ch lng vn ng thch hp ca vn ng vin v kh nng chu ng
ca vn ng vin i vi nng axit lactic cao trong mu l yu t c ngha quyt
nh t c thnh tch cao. Cng tc hun luyn vn ng vin ng ngha vi vic
hun luyn cho vn ng vin tp chu ng ngng axit lactic cao trong mu cc mc
thi gian di, ngn khc nhau
X.1.5. Thiu mu do mt mi trong luyn tp, thi u qu sc v bin php hi
phc.
Ni n thiu mu ca vn ng vin l ni n s gim st mc hemoglobin so
vi nhu cu pht huy nng lc vn ng. Tu theo mc pht trin ca trng thi luyn
tp qu sc m c th t thiu mu ca vn ng vin, h thp xung ti mc ch tiu
thiu mu ca ngi thng v t thiu mu gy gim st nng lc vn ng s dn n
pht trin thnh thiu mu bnh l. Do vy hemoglobin l mt ch tiu quan trng
nh gi th lc ca vn ng vin
122

X.1.6. ng dng cc bi thuc dn gian v c truyn vo vic b sung dinh dng

v thc phm thuc nhm tng cng th lc cho cc vn ng vin Vit nam.
Nng lng l iu kin c bn ca mi hot ng trong c th. Nng lng
khng cc hot ng chc nng gim st, sc kho km; nng lng qu tha li gy
tch m, cng lm cho c th kh vn ng, hot ng chc nng kh khn v sc kho
cng km i. Do vic cung cp thc n hp l l ti quan trng.
Theo kinh nghim c truyn, sao bin l mt hi sn qu ch bin mn n v
lm thuc b dng cho ngi gi, yu, mt mi v tr em chm ln. Sao bin c th
ch bin lm sch, phi hoc sy kh. C th ch bin thnh bt, n cho tht hoc ngm
ru ung. Cng vi cc nghin cu iu tra, phn tch cho thy mt s loi sao bin
nh: Sao bin Gai lc (Comb Sea Star - Astropecten velilaris), Sao ct Spiny (Spiny
Sand Star - Astropecten vappa), Sao Ct Gallopping (Galloping sand star - Stellaster
equestris) rt giu cc hot cht sinh hc nh cc axit amin, cc hoocmn steroit v cc
nguyn t vi lng cn thit cho c th (Nguyn Huy Nam, V Th Ninh, Chu Vn Minh, C
Nguyn nh, v Ngc Quang, Nghin cu thnh phn cc nguyn t vi lng v cc hormone steroid
c trong 3 loi sao bin Vit Nam, Hi ngh khoa hc nhng vn nghin cu c bn trong khoa hc s
sng, 2007). iu ny t c s khoa hc cho vic nghin cu pht trin sn phm thc

phm chc nng tng cng th lc cho cc vn ng vin t sao bin

X.2. kt qu V THO LUN
X.2.1. IU TRA NGHIN CU V THNH PHN CC HOT CHT SINH HC
TRN 3 LOI SAO BIN - Comb Sea Star, Spiny, Gallopping MT S
VNG BIN VIT NAM
Phn tch thnh phn axit amin v cc hoocmn steroit v cc nguyn t vi lng
c trong 3 loi sao bin Vit Nam: Sao bin Gai lc (Comb Sea Star - Astropecten
velilaris), Sao ct Spiny (Spiny Sand Star - Astropecten vappa), Sao Ct Gallopping
(Galloping sand star - Stellaster equestris), chng ti pht hin thy trong 3 loi sao
bin ny rt giu cc hot cht hoocmn steroit, cc nguyn t vi lng.
X.2.1.1. Cc kt qu iu tra phn tch nghin cu v thnh phn axit amin c
trong cc mu nguyn liu th ca 3 loi sao bin: Sao bin Gai lc (Comb Sea
Star - Astropecten velilaris), Sao ct Spiny (Spiny Sand Star - Astropecten vappa),
Sao Ct Gallopping (Galloping sand star - Stellaster equestris).
Cc kt qu nghin cu cho thy trong 3 loi sao bin u rt giu cc axit amin.
Kt qu iu tra phn tch ban u cho thy s xut hin ca 17 axit amin trong loi sao
bin Sao bin gai lc - Comb Sea Star v 16 axit amin trong 2 loi sao bin (Sao ct
piny, Sao ct Gallopping) chng t ngun b sung dinh dng t thc phm sao bin rt
di do v cht lng. c bit l trong loi Sao bin gai lc hm lng axit amin rt
cao, cao nht l Phenylalanin sau n Acginin-Histidin, Glutamic-Treonin .v cui
cng l Histidin.
X.2.1.2. Cc kt qu nghin cu v cc hoocmn steroit c trong mu th ca 3 loi
sao bin: Sao bin gai lc (Comb Sea Star), Sao ct piny (Spiny Sand Star Astropecten vappa), Sao ct Galloping (Galloping sand star - Stellaster equestris)
Cc kt qu phn tch nghin cu v thnh phn hoocmn steroit c trong mu
th ca 3 loi sao bin cho thy hm lng ngun hot cht ny rt cao so vi nhiu loi
ng vt qu him khc.
Hm lng cc hoocmn steroit c trong ngun sao bin s l ngun thc phm
chc nng b sung cho c th rt tt, chc nng v tc dng ca mi loi hot cht nh

123

 ti trnh by phn tng quan ti liu, b sung kch hot h thng min dch,
tuyn thng thn v tng cng th lc cho c th.
X.2.1.3. Cc kt qu nghin cu iu tra v thnh phn cc cht vi khong c
trong mu th thu c ca 3 loi sao bin gai lc - Comb Sea Star, Sao ct
Spiny, Sao ct Gallopping
Qua kt qu nghin cu bng X.2.1.3 cho thy hm lng cc nguyn t vi
lng cn thit cho c th c trong mu th ca 3 loi sao bin cng kh cao so vi cc
i tng ng vt bin nghin cu trc y.
Bng X.2.1.3. Kt qu phn tch hm lng cc vi khong c trong mu th 3 loi sao
bin Sao bin gai lc (Comb Sea Star - Astropecten velilaris), Sao ct piny (Spiny Sand
Star - Astropecten vappa), Sao ct Galloping (Galloping sand star - Stellaster equestris)
Cu
(mg/kg vck)
- Comb Sea Star
41,182,44
Sao ct Spiny
35,295,94
Sao ct Gallopping 34,459,73
Tn mu

Fe
Zn
Mn
(mg/kg vck) (mg/kg vck) (mg/kg vck)
217,2919,95 157,3930,81 36,275,29
208,4833,33 147,8943,10 38,3410,00
236,4551,82 142,2832,21 41,0212,13

Se
(mg/kg vck)
67,8714,56
68,9214,02
62,4815,45

* Gi tr thng k vi: n=30, n = 30, p 0,05

S giu c cc hot cht nguyn t vi khong trong nguyn liu sao bin s l
ngun thc phm v ngun dc liu tt cho ngnh cng nghip ch bin dc v cc
thc phm chc nng.
X.2.2. KT QU NGHIN CU V QUI TRNH TO BT NGUYN LIU CHO
CH PHM BioNAMINE.
X.2.2.1. Kt qu la chn enzym v cc iu kin thy phn
Sau khi iu tra hm lng cc hot cht sinh hc c trong 3 loi sao bin trn,
ti xc nh chn ra loi sao bin Gai lc - Comb Sea Star lm ngun nguyn liu
v hm lng cc hot cht sinh hc c trong sao bin Gai lc - Comb Sea Star cao hn
mt s loi sao bin khc.
Sao bin Gai lc - Comb Sea Star sau khi thu hoch phn loi, chn lc, s ch
xong ngm khong 1-2 gi, ra sch sau cho vo xay nh v bc tip theo l dng
cng ngh enzim ch bin. ti th nghim nghin cu trn loi enzim papain.
Ngun enzym chng ti dng cho nghin cu do Vin Cng ngh Sinh hc cung cp.
la chn loi enyme trn vo vic x l nguyn liu sao bin chng ti lm th
nghim trong qu trnh thu phn trn c s nghin cu d sc xut theo cc iu kin
khc nhau v pH, nhit , thi gian v hot ng c cht. Da trn kinh nghim cc kt
qu nghin cu trc y v cng da trn tnh cht v c th hot ng ca enzim ny
hot ng tt nht l pH trong khong t 6,0 8,0 do vy ti khoanh vng c
tm hot ng ca yu t pH v iu chnh cc yu t nhit , thi gian v t l c
cht.
Qua kt qu thu c cho thy: iu kin pH: 6,5, t0: 400C, Thi gian: 40', T l
c cht E: 0,5%: thu phn sao bin bng enzim Papain cho sn phm c thnh phn
giu axit amin nht c ti 18 axit amin. V iu ny cho thy vic dng enzim papain
thu phn sao bin iu kin trn cho hiu sut ti u. V ti la chn gii php
iu kin nh trn ng dng vo vic sn xut nguyn liu chnh cho vin tng lc
Bionamine v papain l mt enzim c hot tnh proteolytic rt mnh. Nc ta vng
nhit i, c ngun u di do. y l ngun cung cp papain ch yu. Papain d
tch chit, bn vng, gi c hot tnh trong thi gian di iu kin bo qun bnh
thng.
124

X.2.2.2. Kt qu nghin cu so snh v thnh phn axit amin c trong mu bt

nguyn liu sao bin - Sao bin gai lc Comb Sea Star, trc v sau khi thu
phn.
Cc kt qu nghin cu cho thy sn phm sau khi thu phn ca c 3 loi sao
bin v hm lng v s lng lng axit amin u tng ln r rt. iu ny cho thy
kh nng xc tc rt tt ca enzim papain trong qu trnh thu phn, cc protein sao bin
c ct mch thnh cc polypeptit, peptit v cc axit amin.
S xut hin ca 18 axit amin trong loi sao bin bin Gai lc - Comb Sea Star
chng t ngun b sung dinh dng t thc phm sao bin tht di do v cht lng,
hm lng axit amin cao nht l Prolin sau n Tirozin, 4-Hydroprolin,
Aspactic.v cui cng l Treonin.
X.2.2.3. Cc kt qu nghin cu v cc hoocmn steroit c trong mu bt nguyn
liu loi sao bin - Sao bin gai lc trc v sau khi thu phn
Bng X.2.2.3. So snh hm lng cc hot cht steroit c trong mu bt nguyn liu Sao
bin gai lc - Comb Sea Star trc v sau khi thy phn.
Testosteron
(mg/kg cvk)
Mu
Sau TP
th

Tn mu

Sao bin gai

lc

45,02
9,47

62,18
14,04

Cortison
(mg/kg cvk)
Mu
Sau TP
th

47,89
10,16

70,50
15,99

Progesteron
(mg/kg cvk)
Mu
Sau TP
th

47,29
10,42

62,12
13,92

Oestradiol
(mg/kg cvk)
Mu
Sau TP
th

87,93
17,56

97,5
17,37

* Gi tr thng k vi: n=30, n = 30, p 0,05

Cc kt qu phn tch nghin cu bng trn v hm lng hoocmn steroit c

trong mu bt nguyn liu loi sao bin gai lc sau khi thu phn cao hn trc khi
thu phn. iu ny cho chng ta thy cc hoocmn steroit cng u c mt lng d
tr trong v bc t bo. Trong mi t bo lun c mt cu trc v bc bn nhng bnh
thng khng o c. Trong qu trnh thu phn enzim papain c ph v c lp
v bc ca t bo lm bung cc hot cht dinh dng cng nh cc hot cht steroit d
tr bn trong. Do vy sau khi thu phn hm lng cc hot cht steroit c ci
thin tng ln. Hm lng testosteron, Cortison, progesteron v Oestradiol ca loi sao
bin gai lc - Comb Sea Star c tng ln cao. y cng l mt trong nhng ch tiu
ti a ra quyt nh chn ngun sao bin Gai lc - Comb Sea Star lm nguyn
liu chnh cho sn xut vin tng lc Bionamine.
X.2.2.4. Kt qu nghin cu v cc cht vi khong c trong mu bt nguyn liu
loi sao bin - Sao bin gai lc - Comb Sea Star trc v sau khi thu phn.
Bng X.2.2.4. So snh hm lng cc vi khong c trong mu bt nguyn liu loi sao
bin - Comb Sea Star, trc v sau khi thu phn bng papain.
Tn
mu

Sao bin
gai lc

Cu
mg/kg vck
Mu Sau TP
th

Fe
mg/kg vck
Mu Sau TP
th

Zn
mg/kg vck
Mu Sau TP
th

Mn
mg/kg vck
Mu Sau TP
th

Se
mg/kg vck
Mu Sau TP
th

41,18 42,00 217,29 228,93 157,39 162,60 36,27 40,10 67,87 86,80
2,44 2,73 19,95 21,48 30,81 34,08 5,29 11,75 14,56 15,84

* Gi tr thng k vi: n=30, p 0,05

Qua kt qu nghin cu bng trn cho thy hm lng cc nguyn t vi lng

cn thit cho c th c trong loi sao bin sau khi thu phn bng papain cng c ci
thin tng ln so vi trc. iu ny cho thy kh nng ca enzim papain khng ch c
tc dng phn hu cc protein thnh cc axit amin trong qu trnh thu phn m chng
125

cn c tc dng trong vic ph v lp cu trc v bc t bo c cha cc cht dinh

dng v hot cht d tr bn trong, y l lp cu trc bn nhng khng o c, ch
khi c tc dng ca enzim mnh nh papain th lp v t bo b v v cc cht dinh
dng v hot cht d tr bn trong n mi b bung ra. Do vy sau khi thu phn hm
lng d tr cc nguyn t vi lng hu c cn thit trong qu trnh trao i nng lng
ca t bo c trong cc mu sao bin c tng ln. Cc kt qu phn tch trn chng
minh cho thy trong loi sao bin - Comb Sea Star l mt ngun cung cp cc nguyn t
vi lng rt di do cho c th, y cng l mt ngun thc phm chc nng v dc
liu rt tt.
X.2.2.5. qui trnh cng ngh sn xut to bt nguyn liu
Qua cc kt qu nghin cu v mu bt nguyn liu ca 3 loi sao bin cho thy
sau khi thu phn hm lng cc axit amin, cc nguyn t vi lng v hm lng cc
hot cht steroit c ci thin v cao hn trc khi thu phn, c bit l cc mu trong
loi sao bin - Comb Sea Star c hm lng cc hot cht cao hn cc loi sao ct Spiny
v sao ct Gallopping. Do vy ti tin hnh chn loi sao bin Gai lc (Comb Sea
Star - Astropecten velilaris) l nguyn liu cho vic sn xut vin tng lc Bionamine
Thu phn ln 1 (iu kin to, pH, t
l E/c cht v thi gian thch hp)

- Chn lc v s ch nguyn liu

lc
Kim tra hot tnh v hm lng
cc hot cht sinh hc

Thu phn ln 2 (iu kin to, pH, t l

E/c cht v thi gian thch hp)

B sau thu
phn 1

lc

Dch chit ln 1

Dch chit ln 2
trn

Sy phun sng

Kim tra hot tnh v hm lng cc

hot cht sinh hc

Bt nguyn liu
Tinh ch

Hnh X.2.2.5. S qui trnh cng ngh sn xut to bt nguyn liu

X.2.3. KT QU NGHIN CU V C TNH CP TNH V BN TRNG
DIN CA BT bioNAMINE
c tnh ca bt Bionamine c nh gi theo ch tiu: LD50 (liu c gy cht
mt na) v nhim c bn trng din trn ng vt thc nghim. Cc nghin cu thc
nghim v c tnh cp v bn trng din c thc hin ti Trung tm nghin cu c
hc Hc Vin Qun Y. Kt qu nghin cu xc nh LD50 ca hot cht cha trong bt
Bionamine c trnh by trong bng X.2.3.a.

126

Bng X.2.3.a. Bng nghin cu t l sng cht ( xc nh liu c LD50) ca chut

nht trng ung bt Bionamine(n=12)
Mc liu
(mg/g)

Lng
ung
(ml)

S lng
chut
(con)

3,0

0,1

12

12

100

2
3
4
5

7,5
15,0
22,5
30,0

0,25
0,5
0,75*
1,0*

12
12
12
12

12
12
12
12

100
100
100
100

S
TT

Sng/nhm
S
T l
lng
(%)

Ghi ch

Tng vn ng trong khong thi

gian 5-10 pht.
Tng vn ng chung
Tng vn ng chung
Tng vn ng (mc va phi)
Tng vn ng (mc va phi)

* Nhng mc liu c ung lm 2 ln.

Bng X.2.3.b. Kt qu nghin cu c tnh bn trng din ca chut nht trng ung bt
Bionamine (n=12)
L

SL
(chut)

i chng
Xc nh c tnh

25
25

Thng 1
Sng
Cht

25
25

0
0

Thng 2
Sng
Cht

25
25

0
0

Thng 3
Sng
Cht

25
25

0
0

Cn c vo nhng kt qu thu c chng ti iu kin kt lun rng

bt Bionamine khng gy c trc ht l i vi chut v sau l ngi bi l: mc
liu 30 mg/g th trng chut, cc hot cht cha trong bt Bionamine khng gy c
bt k mt biu hin c no trn chut th nghim. Theo tnh ton, y l mc liu cao
hn 1250 ln liu c quy nh s dng trn ngi. Trn thc t kh ai c th tip nhn
c liu cao hn na bi nu theo cch ng gi nh hin nay (300 mg/vin) c
c liu 30 mg/g th trng th mt ngi nng trung bnh 50 kg phi ung s lng
thuc l 5000 vin trong mt ngy m (theo liu quy nh ch phi ung 4 vin/ngy).
Thc nghim bn trng din (kt qu trong bng X.2.3.b) cho thy bt
Bionamine khng gy cht ng vt th nghim trong khong thi gian s dng 03
thng lin tc mc liu 15.000 mg/kg th trng (gp 625 ln s dng cho ngi) khi
xm nhp vo c th qua ng tiu ho.
* Kt lun
1. Bt Bionamine liu 30000 mg/kg th trng khng gy c i vi chut th
nghim. Khng lm cht chut hoc gy xut hin bt k triu chng nhim c no.
2. Vic xc nh chnh xc LD50 ca bt Bionamine trn thc t khng cn thit v
chc chn s cao hn mc liu 30.000 mg/kg th trng nhiu ln, m thm ch ngay
mc liu ny th mt ngi c trng lng trung bnh 50kg cng phi tip nhn trong
vng 24 gi mt s lng 5000 vin Bionamine. iu ny kh c kh nng xy ra trong
thc t.
3. Bt Bionamine khng h gy nh hng bn trng din trn ng vt thc
nghim ngay c khi s dng liu cao, gp 625 ln so vi liu dng cho ngi, trong thi
gian di 3 thng.
X.2.4. KT QU NGHIN CU V QUI TRNH TO VIN TNG LC
BIONAMINE
X.2.4.1. Nghin cu to vin tng lc Bionamine
Da trn kinh nghim v cc bi thuc y hc c truyn, sn phm bt sao bin Gai
lc - Comb Sea Star sau khi thu phn l nguyn liu ca vin tng lc Bionamine c
phi ch vi cc ph gia, t dc theo cng thc v thnh phn nh sau:

127

- Thnh phn hn hp nguyn liu gm bt sao bin Gai lc - Comb Sea Star
thu phn, nhn sm, vitamin B1, B2 v t dc va , trn u. To m cm bng h
tinh bt sy kh. Trn tip vi lng va Talc magnesi stearat c cm.
- em cm ng nang vi khi lng nht nh t hm lng nguyn liu
theo tiu chun (Sai s cho php 64010% mg/vin) vo vin nang, ng 10 vin/v, 4 v
/1 hp.
X.2.4.2. S quy trnh to vin tng lc Bionamine
Lactose, tinh bt

Talc, Mg stearat

Hn hp nguyn liu

To bt kp, sy kh

Cm kh

ng gi

p v

ng nang

Hnh X.2.4.2.a. S qui trnh cng ngh sn xut vin tng lc Bionamine
Sn phm nguyn liu v vin tng lc Bionamine c sn xut trn dy
chuyn thit b hin i t tiu chun ti Cng ty Cng ngh Intecpharm v c
xy dng tiu chun c s v c B Y t cp Chng nhn tiu chun sn phm s
956/2008/YT-CNTC.

Hnh X.2.4.2.b. Chng nhn tiu chun sn phm Bionamine ca B Y t

128

Hnh X.2.4.2.c. Sn phm vin tng lc Bionamine

X.2.5. KT QU NGHIN CU thnh phn v tc dng ca VIN TNG
LC BIONAMINE trn ng vt
X.2.5.1. Kt qu nghin cu v thnh phn axit amin trong vin tng lc
Bionamine
Qua kt qu nghin cu cho thy hm lng cc axit amin c trong sn phm vin
tng lc Bionamine so vi cc sn phm khc trong nc nh Phunamine, Taxaton, ru
Hi m th vin tng lc Bionamine c gi tr cao hn v cng khng thua km g so vi
sn phm vin Triplex hay gi khang thai ca Trung Quc.
Chng ta u bit cc axit amin ng vai tr rt quan trng v thit yu trong
trong c th. Mi axit amin u c nhng ng gp tng chc nng cho c th nh:
A.Aspactic l axit amin c tc dng gip cho qu trnh gii c v tng cng sc bn
c th. A.glutamic b no, gip mau lnh vt thng, kch thch hng phn thn kinh.
Treonin kch thch h thng thn kinh thc vt gip cho ng ngon tng cng chc nng
gan. Xerin tng cng min dch, sc khng v tng sc bn. Glyxin tng c bp,
gip tuyn tin lit kho mnh. Histidin c tc dng b khp, tng to mu chng d
ng. Metionin, Valin gip cho n nh tm l. PhenylAlanin c tc dngngn nga
chng bo ph, gim trm cm, gim au nhc do bnh cm, kinh nguyt, gim vim
khp. Izolxin l axit amin thit yu n nh lng ng trong mu, tng cng h tr
pht trin c bp.
Vi thnh phn axit amin ca vin tng lc Bionamine so vi cc sn phm
tng t khc trong nc v nc ngoi khng km g v gi tr chc nng dinh dng
do vy sn phm rt c tin dng cho vic nng cao th lc ca vn ng vin l rt tt.
X.5.5.2. Nghin cu thnh phn cc cht vi khong trong vin tng lc Bionamine
V hm lng cc hot cht vi khong sau khi bo ch vin tng lc Bionamine
Qua kt qu nghin cu trong bng X.2.5.2. cho thy hm lng cc cht vi khong c
trong sn phm vin tng lc Bionamine so vi cc sn phm tng t trong nc cao
hn v so vi sn phm gi khang thai v vin Triplex ca Trung Quc th kt qu cng
khng khc nhau nhiu nh ca Bionamine, thm ch mt s thnh phn ca hm lng
nguyn t v st, km hay selen cn cao bng hoc hn cc sn phm ni v ngoi.

129

Bng X.2.5.2. Hm lng cc cht vi khong c trong vin Bionamine so vi cc sn

phm khc.
Hm lng vi
cht
Cu(mg/kg vck)
Fe (mg/kg vck)
Zn (mg/kg vck)
Mn(mg/kg vck)
Se(mg/kg vck)

Vin
Bionamine
4,20
228,93
16,26
40,10
86,80

Vin
Phunamine
3,22
231,08
7,27
32,90
-

Vin
Taxaton
(VN)
4,01
227,20
6,26
27,24
-

Gi Khang
thai (TQ)
6,22
240,02
11,28
37,91
-

Vin
Triplex ca
(TQ)
3,92
238,27
3,24
42,76
-

Vi thnh phn cc cht vi khong hu c c trong vin tng lc Bionamine rt

phong ph c bit l nguyn t st s gip cho qu trnh tng to hng cu, hemoglobin
trong c th rt tt. Hm lng ng, mangan c trong vin Bionamine gip cho s pht
trin tr tu, kch thch phn x thn kinh c tt hn, hm lng km c trong vin
Bionamine cao s gip cho qu trnh tng cng th lc, tng cng min dch gip c
th thc hin tt chc nng ni tit, tun hon mu, mau lnh vt thng, khng ch t
bo gy nhiu. c bit nguyn t km cn ng vai tr chc nng duy tr sinh l ho
trong c th, tng kh nng sinh dc cho nam gii, gim bt cc nguyn t vi lng t
do. kch thch cc hoocmn tuyn thng thn hot ng.
X.2.5.3. Nghin cu thnh phn cc hot cht hoocmn steroit trong vin tng lc
Bionamine
V hm lng cc hot cht steroit, sau khi bo ch vin tng lc Bionamine Qua
kt qu nghin cu trong bng 3.5.3 cho thy hm lng cc hot cht steroit c trong
sn phm vin tng lc Bionamine cng kh cao so vi cc sn phm khc, k c sn
phm ngoi, iu ny chng t sn phm trong nc nu c ng dng khoa hc cng
ngh th cht lng v gi tr s c nng ln r rt. Hm lng Testosteron (mg/kg
vck) c trong vin Bionamine l (69,02) cao hn vin Taxaton (68,00) v gi khang thai
ca Trung quc (67,35) nhng cao hn vin Triplex ca Trung quc (32,08).
Bng X.2.5.3. Hm lng cc hot cht steroit c trong vin Bionamine so vi cc sn
phm khc.
Hm lng steroit

Vin
Bionamine

- Testosteron(mg/kg vck)
- Progesteron(mg/kg vck)
- Cortisol (mg/kg vck)

69,02
79,73
56,02

Ru
Hi m
(VN)
11,02
90,28
8,22

Vin
Taxaton
(VN)
68,00
65,07
19,40

Gi
Khang
thai (TQ)
67,35
52,15
59,24

Vin
Triplex
ca (TQ)
32,08
38,72
55,12

Hm lng Progesteron(mg/kg vck) ca vin Bionamine l (79,73) cao hn nhiu

so vi cc sn phm trong nc v ca Trung Quc nh ru Hi m (9,28), vin
Taxaton (65,07), gi khang thai (52,15) v vin Triplex (38,72). Hm lng Cortisol
(mg/kg vck) ca vin Bionamine l (56,02) cao hn tt c cc sn phm trong nc nh
ca ru Hi m (8,22), vin Taxaton (19,40) v khng thua km g cc sn phm ca
Trung Quc nh gi Khang Thai (19,40), vin Triplex (55,12).
Vi thnh phn v hm lng cc hot cht hoocmn steroit c trong vin tng
lc Bionamine c tc dng ln cc chuyn ho trung gian, gluxit, lipit v protein. Cc
hoocmn steroit kch thch tng hp lipoprotein gip cho vic b p v cn bng
glucose v lipit trong mu. Hoocmn steroit vi nng cao c tc dng tng cng
min dch, chng vim nhim, chuyn ho mui ln tuyn nc bt v tuyn m hi

130

cng nh s vn chuyn mui v nc qua nim mc ng tiu ho, tham gia vo s duy
tr cn bng nc v in gii m bo s vn ng tiu hao nng lng c th.
X.2.5.4. Kt qu nghin cu v th nghim tc dng ca vin tng lc Bionamine
trn ng vt thc nghim.
a. Tc dng ln mt s ch tiu huyt hc ca chut cng trng ca vin tng lc
Bionamine.
Bng X.2.5.4..a Bng kt qu kim tra cc ch tiu huyt hc ca chut dng th nghim
ch phm.
L Th nghim dng Vin Bionamine
Tn
CT

WBC
(x103/l) S
lng tuyt i
bch cu oan
trung tnh

RBC (x106/l)
S lng hng
cu

Trc

Sau

Trc

Sau

Trc

2,30

2,00

3,33

4,04

SD
0,22
0,22
0,48
0,67
* Gi tr thng k vi: n = 10, p 0,05

HGB
(g/dL)
Huyt sc t

Sau

L i chng dng Vin Placebo

WBC (x103/l)
S lng tuyt
i bch cu
oan trung tnh

RBC (x106/l)
S lng hng
cu

HGB (g/dL)
Huyt sc t

Trc

Sau

Trc

Sau

Trc

Sau

12,76 14,02

2,00

2,10

3,34

3,39

12,78

12,67

0,55

0,18

0,16

0,43

0,46

0,73

0,68

0,84

Kt qu bng trn cho thy sau khi cho chut cng trng n b sung vin tng
lc Bionamine trong 20 ngy th lm thay i mt s ch tiu huyt hc quan trng
ca chut.
Qua kt qu th nghim trn cho thy vin tng lc Bionamine thc s c tc
dng kch thch s to mu, to hng cu v hemoglobin. S tng s lng hng cu c
ngha tng cng cung cp oxy cho t bo, iu rt cn thit cho qu trnh vn ng
v cc hot ng ca c bp cng nh vic tng cng th lc ca c th.
b. Tc dng tng cng th lc v sc bn ca ch phm Bionamine trn chut trong
qu trnh bi ln.
Bng X.2.5.4.b. Bng kt qu kim tra thi gian bi ca chut.
Chut TN
Chut ci
Chut c

L i chng
TrcTN
Sau (TN) 15ngy
3,36
3,41
3,25
3,35
3,31
3,38

X
* Gi tr thng k vi: n = 10, p 0,05

L (TN1) B sung Bionamine

TrcTN
Sau (TN) 15 ngy
3,26
4,07
3,28
4,02
3,27
4,05

Qua kt qu bng trn cho thy gia cc l i chng v th nghim c s khc

nhau r rt, thm ch s khc nhau cn th hin c kt qu trong cc l th nghim t
i chng.
- l i chng thi gian bi trung bnh ca chut gia trc th nghim l 3,31 v sau
t th nghim l 3,38 kt qu c s khc nhau khng nhiu lm.
- l th nghim (TN) chut th nghim c cho n b sung ch phm Bionamine v
thi gian bi trung bnh c ci thin tng c 0,38 trc th nghim l 3,27, sau th
nghim kt qu l 4,05 .
Cc kt qu thc nghim trn chng minh cho thy cc ch phm Bionamine thc
s c tc dng tng cng th lc i vi chut th nghim th hin thi gian bi ca
chut c ko di hn, sc chu ng ca chut c tt hn. iu ny l gii rng
trong cc ch phm cung cp b sung thm cc cht dinh dng cng nh cc hot
cht sinh hc, cc hoocmn steroit (testosteron, progesteron), cc cht vi khong nh
131

Fe, Zn, Cu..v cc axit amin cn thit trong qu trnh vn ng v tiu hao nng lng
ca chut.
T cc thc nghim chng minh c tc dng ca vin tng lc Bionamine i
vi sc bn v tng cng th lc trn chut, ti tin hnh trin khai thc nghim
trn vn ng vin mn in kinh, bng bn, mn c t v thu c cc kt qu
di y.
X.2.6. Kt qu nghin cu v th nghim vin tng lc Bionamine
trn vn ng vin
X.2.6.1. nh gi v tc dng sc bn trn vn ng vin
nh gi c sc bn ca vn ng vin in kinh ti tin hnh ly mu
mu v kim tra cc ch tiu sinh ho lin quan n sc bn ca th lc vn ng vin v
kt qu cho thy:
a. Tc dng hi phc v nng cao hm lng Testosteron trong mu vn ng vin
ca vin tng lc Bionamine.
Kt qu bng X.2.6.1.a cho thy hm lng testosteron ca VV nhm i
chng (khng dng vin Bionamine) sau 1 thng tp luyn v thi u: VV nam
gim 3,62% t 24,28 nmol/l xung 23,40nmol/l, cn VV n gim 11,93%, t 2,18
nmol/l xung cn 1,92 nmol/l.
Bng X.2.6.1.a. Hm lng Testossterone trong mu ca VV in kinh trc v sau 1
thng luyn tp.
Hm lng
Testossterone (nmol/l)
Nam VV

SD
N VV

SD

Nhm C dng placebo

Sau 30 ngy
Trc t tp
tp
24,28
23,40
3,71
3,84
2,18
1,92
0,49
0,26

Nhm TN dng Bionamine

Trc t
Sau 30 ngy
tp
21,92
24,95
1,92
1,83
1,46
2,04
0,44
0,39

* Gi tr thng k vi: n = 14, p 0,05

nhm th nghim dng vin tng lc Bionamine sau 1 thng tp luyn v thi
u hm lng testosteron nh sau: Vn ng vin nam trc khi dng v sau khi dng
tng 13,84%, t 21,92 nmol/l n 24,95 nmol/l. Nhm vn ng vin n sau 1 thng
dng vin tng lc Bionamine hm lng testosteron tng t 1,46 nmol/l ln
2,04nmol/l.
Cc kt qu trn cho thy rng vin tng lc Bionamine gp phn ci thin hm
lng testosterrone hao ht t nhin trong c th sau t tp luyn, gip cho vn ng
vin hi phc nhanh hn.
b. Tc dng b sung b p nng cao hm lng cortisol trong mu vn ng vin
tc ng tch cc vo qu trnh nng cao th lc v hi phc sc kho cho vn ng
vin ca vin tng lc Bionamine.
Cortisol (hoocmn thuyn thng thn) l mt tham s quan trng trong vic
nh gi lng vn ng ca c th. Khi c th vn ng nng s sinh ra s tiu hao
nng lng ln
Kt qu bng X.2.6.1.b. cho thy hm lng cortisol trong mu gia cc nhm
i chng v nhm nghin cu dng vin tng lc Bionamine c s khc nhau r rt.
- Vn ng vin n nhm i chng sau mt thng tp luyn khng dng b
sung vin Bionamine hm lng cortisol b hao ht t 542,25 (nmol/l) xung cn
132

530,25 (nmol/l). Trong khi nhm nghin cu c dng b sung vin tng lc
Bionamine sau 1 thng tp luyn hm lng cortisol trong mu khng nhng khng b
gim m cn c b p tng thm t 540,17 (nmol/l) ln 544,00 (nmol/l).
Bng X.2.6.1.b. Hm lng Cortisol trong mu ca VV in kinh trc v sau 1 thng
luyn tp
Hm lng Cortisol
(nmol/l)
Nam VV

SD
N VV

SD

Nhm C dng placebo

Trc t tp

Sau 30 ngy tp

545,93
57,12
542,25
78,05

536,55
52,69
530,25
72,15

Nhm TN dng Bionamine

Trc t
Sau 30 ngy
tp
541,55
584,42
51,05
64,07
540,17
544,00
75,46
96,53

* Gi tr thng k vi: n = 14, p 0,05

- Vn ng vin nam nhm i chng sau mt thng tp luyn khng dng b

sung vin Bionamine hm lng cortisol cung b gim t 545,93(nmol/l) xung cn
536,55(nmol/l). Trong khi nhm nghin cu c dng b sung vin tng lc
Bionamine sau 1 thng tp luyn hm lng cortisol trong mu tng thm t
541,55(nmol/l) ln 584,42(nmol/l). Cc s liu so snh trn u mc cho php p<0,05
do vy kt qu nghin cu l hon ton c gi tr.
T vn hm lng cortisol t nhin c trong thnh phn ca vin tng lc
Bionamine cao. Thng qua vic dng b sung vin tng lc Bionamine cho vn ng
vin in kinh nghin cu trn chng minh rng vin tng lc Bionamine b sung rt
tt v kp thi vo qu trnh hao ht cortisol gim st trong qu trnh vn ng. Khc
vi nhng hoocmn protein, cortisol cng nh testosteron l nhng hoocmn steroit
khng c d tr trong nhng nang bi tit v cng khng c hoocmn d tr trong
nang t bo. Cortisol cng nh testosteron c th khuch tn t do qua mng, t bo v
vo mu. trong tun hon mu, phn ln cortisol cng nh testosteron v cc
hoocmn steroit kt hp vi protein c hiu ca huyt tng, bng cch ny cortisol
cng nh cc hoocmn steroit khc tn ti t do trong tun hon lu hn so vi dng
steroit t do. Ch c cc hoocmn steroit t do l c tc dng hot ng sinh hc. Vic
b sung cortisol cng nh testosteron v cc hoocmn steroit bng vic dng vin tng
lc Bionamine c tc dng rt tt trong vic tng cng th lc v hi phc sc kho ca
vn ng vin.
c. Tc dng b tr chc nng gan trong c th, kch hot enzym Lactat
Dehydrogenaza (LDH) gip tng nhanh qu trnh hi phc ca vin tng lc
Bionamine.
Qua kt qu bng X.2.6.1.c cho ta thy vn ng vin cc nhm i chng sau
1 thng tp luyn khng dng vin tng lc Bionamine hot LDH trong huyt thanh
hu nh khng thay i hoc tng ln khng ng k t 0,61-4,52%. vn ng vin n
thay i trc tp luyn v sau tp luyn t 398,40 UI/l ln 416,40 UI/l (tng 4,52% ).
Cn vn ng vin nam thay i trc tp luyn v sau tp luyn t 390,60UI/l ln
393,00 UI/l (tng 0,61% ).
Trong khi nhm thc nghim c c dng b sung vin tng lc Bionamine
trong thi gian tp luyn, hot LDH nam VV tng t 428,47 UI/l n 478,27 UI/l
(11,62%), n VV tng 11,63% (t 381,13 UI/l n 425,47 UI/l), iu ny c th gii
thch l do vin Bionamine c thnh phn v hm lng km hu c cao. Trong nhng
cng trnh nghin cu mi gn y khng nh rng Zn km c tc dng kch hot
enzym LDH trong phn ng kh axit lactic to thnh axit piruvic, y l mt yu t rt
cn thit i vi cc vn ng vin v cc i tng lao ng c bp.
133

Bng X.2.6.1.c. Hm lng Lactat Dehydrogenaza (LDH) trong mu ca VV in kinh

trc v sau 1 thng tp luyn.
Hm lng LDH
trung bnh (UI/l)
Nam VV

SD
N VV

SD

Nhm C dng Placebo

Trc t
Sau 30 ngy
tp luyn
tp luyn
390,60
33,11
398,40
21,30

393,00
65,15
416,40
40,27

Nhm TN dng Bionamine

Trc t
Sau 30 ngy
tp luyn
tp luyn
428,47
478,27
36,93
381,13
43,32

53,52
425,47
31,97

* Cc gi tr thng k vi n=14 v p0,05

d. Tc dng trong qu trnh phn gii axit lactic trong c lm tng cng kh nng
hi phc ca c th vn ng vin ca vin tng lc Bionamine.
Bng X.2.6.1.d. Hm lng axit lactic trong mu ca VV in kinh trc v sau 1
thng tp luyn.
Hm lng axit
lactic trung bnh
(mg/dl)
Nam VV
X
SD
N VV
X
SD

Nhm C dng placebo

Sau 30 ngy
Trc t tp
tp
4,26
4,49
0,34
0,30
3,96
4,29
0,18
0,24

Nhm TN dng Bionamine

Sau 30 ngy
Trc t tp
tp
3,24
2,60
0,40
0,25
4,02
2,74
0,21
0,50

* Cc gi tr thng k vi n=14 v p0,05

Cc kt qu trong bng X.2.6.1.d cho thy cc vn ng vin nhm i chng

khng dng vin tng lc Bionamine, hm lng axit lactic nam VV sau t tp
luyn khng c g thay i m c v tng nh, tng t 4,26 mg/dl ln 4,49 mg/dl (5.4%),
cn n vn ng vin nhm i chng khng dng Bionamine hm lng axit lactic
trung bnh tng t 3,96 mg/dl ln 4,29 mg/dl (8,28%). iu ny cng rt c l v sau
mi t tp luyn vn ng vin thng mt mi, nguyn nhn gy mt mi chnh l
Axit lactic tn ng li nhiu trong c, nu c xc tc nhiu ca enzim LDH th axit
lactic se c phn hu v s lm gim mt mi, khng b co c.
Cc vn ng vin nhm th nghim c dng vin tng lc Bionamine, hm
lng axit lactic trung bnh nam VV sau t tp luyn c ci thin gim t 3,24
mg/dl xung 2,60 mg/dl (19,63%), n VV nhm th nghim gim 31,83% (t 4,02
mg/dl xung 2,74 mg/dl) cc kt qu ny cho thy vin Bionamine c tc dng kch hot
enzym LDH tng cng phn ng kh axit lactic to thnh axit piruvic, y l mt yu
t rt cn thit i vi cc vn ng vin v cc i tng lao ng c bp.
e. Tc dng hi phc v tng hng cu v ci thin cc ch tiu huyt hc ca vn
ng vin ca vin Bionamine.
Kt qu th nghim cho thy th tch trung bnh hng cu (MCV) ca cc VV
nhm th nghim sau khi dng vin tng lc Bionamine mc n nh tt: nam VV
t 88,05 f/l n 89,13 f/l v n VV t 88,68 f/l n 90,10 f/l.
S lng bch cu WBC ca VV nhm th nghim trc v sau t tp luyn
c dao ng nhng mc n nh tt: Nam VV t 6,57 (x103/l) n 6,29 (x103/l) v
n VV t 6,13 (x103/l) n 5,91 (x103/l).
Huyt sc t (HGB) cc VV th nghim u gi c mc n nh: nam
VV t 14,00 g/dl n 14,69 g/dl v n VV t 12,21 g/dl n 12,86 g/dl.
S lng hng cu nhm VV th nghim sau 1 thng tp luyn c dng vin
tng lc Bionamine cho thy s lng hng cu khng nhng khng b gim m cn
134

c ci thin tng ln, nam VV hng cu tng 4,98 (x106/l) ln 5,56 (x106/l),
n VV hng cu tng t 4,29 (x106l) ln 4,90 (x106/l).
Cc kt qu tng t nhn c trong thc nghim vi cc VV nhm i
chng khng dng vin tng lc Bionamine trong t tp luyn nh sau: Hng cu
nam VV c thay i khng nhng khng tng m cn gim nh t 4,82 (x106/l) n
4,62 (x106/l) v n VV tng t 4,39 (x106/l) n 4,26 (x106/l).
Huyt sc t (HBG) nam VV nhm i chng gim t 13,76 g/dL n 13,32
g/dL, n VV gim t 12,26 g/dL n 11,76 g/dL.
S lng bch cu WBC ca VV nhm i chng trc v sau t tp luyn c
dao ng theo nh hng ca s lng hng cu: Nam VV t 6,92x103/l n
7,02x103/l v n VV t 6,72x103/l n 6,83x103/l.
Th tch trung bnh hng cu (MCV) ca cc VV nhm VV i chng khng
dng vin tng lc Bionamine sau t tp luyn hu nh khng c s thay i: nam
VV t 87.64 f/l n 87.06f/l v n VV t 87.06 n 86.84 f/l.
f. Tc dng hiu qu ln cc test gng sc ti a ca VV ca vn ng vin in
kinh ca vin tng lc Bionamine.
Thc nghim c tin hnh kim tra nh gi trn cc vn ng vin in kinh
trc v sau 1 thng tp luyn c dng th nghim vin tng lc Bionamine. Kt qu thu
c trn thit b chn on chc nng Cosmed ca Italia cho 20 VV in kinh (10
nam v 10 n). Kt qu thu c cho thy nhng vn ng vin nam c dng vin vin
tng lc Bionamine trc v sau 1 thng luyn tp cc test kim tra u cho cc kt qu
r nt hn.
Bc cng sut VV thc hin c l c s chnh nh gi kh nng gng sc.
Test gng sc c thc hin trn xe p lc k theo bc cng sut tng dn. Nam gii
mi bc cng sut tng 50W, thi gian mi bc cng sut ko di 2 pht. Kt qu cho
thy cng sut vn ng thc t trc khi th nghim t c l 150W, t 95% so vi
d bo, sau vn ng thc t t 190W t 109% so vi d bo. Tng 14%. n mi
bc cng sut tng 30W, thi gian mi bc cng sut ko di 2 pht. Kt qu cho thy
cng sut vn ng thc t trc khi th nghim t c l 150W, t 86% so vi d
bo, sau vn ng thc t t 180W t 95% so vi d bo. Tng 9%.
200
Sau TN , 190
180
160
Trc TN, 150
140
120

Sau TN , 100

100
Trc TN, 91
80
60
Sau TN , 43.7
40
Trc TN, 33.3
20

Sau TN , 14.2
Trc TN, 13.5

0
Cng sut vn ng load (W)

Sc bn a kh VO2max/Kg (ml/ph/Kg)

Chc nng h hp VE(lt)

Chc nng tim mch VO2/HR (ml/m)

Hnh X.2.6.1.a. Biu so snh test gng sc vn ng vin nam i in kinh

H Ni thng qua cc ch s chc nng trc v sau
thi gian dng vin tng lc Bionamine.
Vn ng vin nam: Sc bn a kh gia trc v sau tng 10.4 (ml/ph/Kg),
chc nng h hp kh nng d bo 17%. Kh nng vn chuyn oxy ca h tim mch
c nh gi thng qua ch s oxy/mch (VO2/HR) l th tch oxy c tim bm vo
mu sau mi ln tim co bp. n v tnh l ml/mch p (ml/m), chc nng tim mch
135

gia trc v sau thc nghim tng 0.7(ml/m). vn ng vin n: Sc bn a kh gia

trc v sau tng 9,4(ml/ph/Kg), chc nng h hp tng gia trc v sau thc nghim
l 31lit kh nng d bo tng sau thc nghim l 60% tng 30%. Kh nng vn chuyn
oxy ca h tim mch c nh gi thng qua ch s oxy/mch (VO2/HR) l th tch
oxy c tim bm vo mu sau mi ln tim co bp. n v tnh l ml /mch p
(ml/m), tng gia trc v sau thc nghim l 3,8(ml/m). Cc ch s ny cho thy sau
thc nghim l rt tt.
180

Sau TN , 180

160
Trc TN, 150
140
120
100
Sau TN , 75

80
60

Trc TN, 44

Sau TN , 37.4

40

Trc TN, 28
Sau TN , 15
Trc TN, 11.2

20
0
Cng sut vn ng load (W)

Sc bn a kh VO2max/Kg
(ml/ph/Kg)

Chc nng h hp VE(lt)

Chc nng tim mch VO2/HR

(ml/m)

Hnh X.2.6.1.b. Biu so snh test gng sc vn ng vin n i in kinh H ni

thng qua cc ch s chc nng trc v sau thi gian dng
vin tng lc Bionamine.
Cc kt qu trn chng minh rng sau 1 thng dng vin tng lc Bionamine cc
ch s test th lc ca vn ng vin nam v n u c ci thin r rt.
g. Tc dng tng cng th lc v nng cao thnh tch ca vn ng vin in kinh
ca vin tng lc Bionamine.
Thc nghim c chng minh trn 48 VV in kinh c chia 2 nhm (C v
TN), mi nhm li c chia theo nhm nam v n. Nhm TN dng vin tng lc
Bionamine, nhm C dung vin Placebo. Thi gian theo di l 30 ngy thc nghim.
Bng X.2.6.1.e. Kt qu kim tra thnh tch ca cc VV trc v sau khi dng vin
tng lc Bionamine. (n=14, p0.05)
Ch tiu

Nhm VV
VV TN
(N): X
VV
C(N): X
VV
TN(Nam): X
VV C
(Nam): X

C ly 100 m (s)
C ly 200m (s)
C ly 400m (s)
Trc
Sau
SS
Trc
Sau
SS
Trc
Sau
SS
TN
TN 30
TN
TN 30
TN
TN 30
ngy
ngy
ngy
13,29 12,92
0,37 26,43 26,01 0,42 57,92 56,9
1,02
0,10 0,10 0,01 0,26 0,27 0,04 0,24 0,24 0,01
13,27 13,26
0,01 26,13 26,10 0,03 56,98 56,94
0,04
0,04 0,04 0,00 0,06 0,07 0,01 0,12 0,13 0,01
11,84 11,46
0,38 22,53 22,08 0,45 51,18 49,75
1,43
0,14 0,14 0,00 0,16 0,11 0,05 0,27 0,25 0,08
11,88 11,84
0,03 22,55 22,52 0,03 50,54 50,50
0,04
0,04 0,03 0,01 0,01 0,01 0,01 0,12 0,12 0,01

Kt qu trong bng X.2.6.1.e cho chng ta thy hu ht cc VV dng vin tng

lc Bionamine thnh tch u c ci thin r rt (C ly 100m: VV n tng trung
bnh 0"37, VV nam tng trung bnh 0"38. C ly 200m VV n tng trung bnh 0"42,
VV nam tng trung bnh 0"45. C ly 400m VV n tng trung bnh 102, VV nam
tng trung bnh 143. Trong khi VV cc nhm C hu nh khng c s thay i.

136

iu vin tng lc Bionamine thc s c tc dng tng cng b sung cc hot cht
kch thch tng cng th lc, nng cao thnh tch ca VV.
X.2.6.2. nh gi v tc dng tng cng thn kinh phn x tr nh trn vn ng
vin
Bng X.2.6.2.a. Kt qu kim tra phn x ca VV bng bn - i hc TDTT T Sn
Ch tiu
Nhm
VV
nam

VV
n

MaX
Min
X

SD
MaX
Min
X

SD

Phn x n
Ln 1
Ln 2
(ms)
(ms)
227,84
176,20
155,84
95,00
192,15
131,70
21,34
25,14
256,18
227,69
190,39
136,92
223,97
178,19
21,26
26,00

Ln 1
(ms)
352,97
245,51
306,38
29,25
421,00
190,39
268,05
59,25

Phn x phc
Ln 2
S li
(ms)
Ln 1
276,00
5,00
165,00
1,00
204,38
3,00
28,08
1,17
219,84
6,00
133,15
1,00
167,02
3,38
21,52
1,17

S li
Ln 2
3,00
0,00
0,81
0,81
3,00
0,00
0,88
0,78

Gi tr thng k sinh hc vi n=14, p0,05

* (ms): miligiy
* Ln 1: Kim tra trc t tp luyn.
* Ln 2: Kim tra sau 1 thng tp luyn

Kt qu kim tra phn x trong bng X.2.6.2.a cho thy test kim tra phn x
n cc VV nam trc v sau thi gian tp luyn 1 thng trung bnh t 192.15ms
xung 131.70ms gim 31%. Cn vn ng vin n gim trung bnh t 223.97ms
xung 178.19ms gim 20.44%.
Phn x phc ca vn ng vin nam sau 1 thng dng vin tng lc Bionamine
c ci thin trung bnh t 306.38ms xung 204.38ms gim 33.29%. Cn vn ng
vin n phn x phc t 268.05ms xung cn 167.02ms gim 37.69%.
S li mc phi trong phn x phc ca VV nam c ci thin trung bnh t 3
ln xung cn 0.81 ln. VV n gim trung bnh t 3.38 ln xung cn 0.88 ln.
Cc kt qu kim tra v phn x n, phn x phc v s ln mc li in t gia
trc v sau khi dng h tr vin tng lc Bionamine c s khc bit nh trn ta c th
nh gi c tc dng ca vin tng lc Bionamine gip cho cc vn ng vin tng
hiu sut vn ng, kch thch hot ng ca thn kinh trung ng x l tt phn x
trong cc bi tp.
X.2.7. tho lun
Mi qu trnh trao i cht u thng qua hot ng ca h thng enzym v chu
s phi hp iu ho ca h thn kinh, ni tit, v h iu tit t ng trong t bo
(iu khin cc phn ng ph hp vi yu cu ca c th). C th con ngi l c mt
h thng phc tp, c hng trm nghn cc phn ng ho hc khc nhau. H thng ny
hot ng rt chnh xc, ng b, nhanh nhy theo cc tc ng mi trng bn ngoi
v bn trong ca c th. cc phn ng trong c th hot ng c lp hay lin kt v to
thnh mt qu trnh. l nn tng ca s sng. Nhng tc ng lm bin i qu c
cht v thnh phn ni mi trong c th s c iu chnh bi tc ca cc phn ng
ho sinh trong mt gii hn nht nh. Trong iu kin thnh phn ni mi ca c th
khi qu c cht b ph hu v gim xung th mi s vn ng hay iu tit s t ng
iu tit cn bng. Vn ny c chng minh bng cc thc nghim trn vn ng
vin trong iu kin vn ng cng cao, khi thnh phn ni mi b ph hu v qu
137

c cht b gim xung sau 1 thng tp luyn v thi u. Hiu qu phc hi , thm ch
phc hi nhanh 1 s cc ch tiu v th lc cc vn ng vin di nh hng tc dng
ca vin tng lc Bionamine. iu ny th hin cc im di y:
- Cc hot cht sinh hc c trong thnh phn ca vin tng lc Bionamine
cung cp v b sung kp thi cho c th cc ngun hocmon Testosteron t nhin, m
khng phi l tng hp ho hc. Vai tr ca Testosteron gip cho c th tng thc
y sinh tng hp protein, tng tch lu glucogen, y mnh chuyn ho nit,
photpho..v..v..
- Vin tng lc Bionamine cung cp cho c th sn phm giu cht vi khong
dng lin kt hu c (Fe, Zn, Cu) nhm tng to mu, thc y sinh tng hp
hemoglobin, feritin, cc enzym h hp v trao i cht.
- Vin tng lc Bionamine cung cp cho c th VV cc hot cht c hot
tnh sinh hc c tc dng tng hot ho enzym LDH gii phng axit lactic v do c
th nng cao c ngng chu ng axit lactic, tng nhanh qu trnh hi phc ca c
th. Vin tng lc Bionamine c tc dng tng cng cng sut vn ng, chc nng
tim mch v h hp gip cho vn ng vin tng cng th lc tt hn.
X.3. Kt lun.
1. Cc kt qu nghin cu trn chng minh cho thy trong ngun nguyn liu 3 loi
sao bin - Comb Sea Star, Spiny, Gallopping rt giu cc hot cht axit amin, cc nguyn
t vi lng v hot cht hoocmnr steroit cn thit v c gi tr b dng cho sc kho
con ngi.
2. Vic ng dng cng ngh ho sinh enzim v s dng enzim papain thu phn
lm giu thm hm lng cc hot cht sinh hc c trong nguyn liu loi sao bin - Sao
bin gai lc (Comb Sea Star - Astropecten velilaris). lm cho hm lng cc hot
cht axit amin, cc nguyn t vi lng v hot cht hoocmn steroit l cao nht. iu
ny chng t cho thy sao bin Gai lc l ngun nguyn liu qu cho ngnh ch bin
thc phm chc nng v cng nghip dc.
3. Sn phm bt nguyn liu Bionamine qua thc nghim trn ng vt cho thy an
ton v c tc dng y l thnh phn rt tt ca vin tng lc Bionamine, l nhng
ngun thc phm chc nng v dc liu c gi tr. Th hin cc hm lng axit amin,
hoocmn steroit v cc nguyn t vi lng l cao so vi mt s loi sn phm tng t
khc trong nc v nc ngoi.
4. Vin tng lc Bionamine an ton v c tc dng thc s r nt thng qua vic
thc nghim trn chut.
5. Vin tng lc Bionamine thc s c tc dng i vi vn ng vin in kinh
trong vic hon thnh tt cc bi tp th lc, kch thch phn x thn kinh hot ng tt
hn, tng cng c cc t cht v sinh l, sinh ho cho vn ng vin v nng cao
c thnh tch tp luyn ca cc vn ng vin.
6. Vic ng dng vin tng lc Bionamine trong vic tng cng th lc v hi phc
nhanh sc kho cho vn ng vin nhm nng cao thnh tch trong tp luyn v thi u
l rt ph hp v c hiu qu mang tnh mi v khoa hc v cng ngh.
Vin tng lc Bionamine qua thi gian thc nghim cho thy s dng an ton v yn
tm trn vn ng vin, khng c phn ng ph xy ra. ngh B Vn ho - Th thao
v Du lch cn c trin khai dng rng ri cho vn ng vin ti cc gii thi u trong
nc v quc t nhm tng cng th lc v hi phc sc kho nng cao thnh tch ca
nn th thao nc nh.

138

Chng xi. Nghin cu bo ch v nh gi tc dng bi b

c th ca vin nang bo ng (Haliotis)
xI.1. m u
Phn ny gii thiu tm tt nghin cu pht trin sn phm t ngun dc liu
bin, tim nng ca Vit Nam trong lnh vc ny, c s v mc ch la chn nghin
cu sn xut sn phm thc phm chc nng HALIOTIS t bo ng.
XI.2. kt qu v tho lun
XI.2.1. Kt qu nghin cu qui trnh bo ch vin nang HALIOTIS
t Bo ng
Bo ng

X l sch tp cht

Sy kh (50-60o)

Nghin bt mn

Trn t dc

ng nang mm

Sy nang

ng l, p v, hon thin sn
phm
Kim nghim
thnh phm
Nghin cu

Hnh XI.2.1.a. S quy trnh sn xut vin nang mm bo ng

- Nghin cu la chn dc liu v xy dng qui trnh bo ch vin nang mm Bo
ng Haliotis thnh dng bn thnh phm v thnh phm: xy dng c qui trnh
bo ch sn phm bo ng di dng nang mm nh sau:

139

Cng thc cho 1 vin:

Bt kh bo ng: 150 mg
T dc v:
01 vin
Tin hnh sn xut th nghim c 67.000 vin nang trn dy chuyn sn xut
nang mm hin i lm sn phm mu v phc v cho cng tc p dng trn lm sng
v ng k lu hnh sn phm.
- Nghin cu xy dng tiu chun c s cho sn phm vin nang mm bo ng
Haliotis: tin hnh nghin cu xy dng tiu chun c s ph hp vi ch tiu do B
Yt qui nh cho sn phm.
+ Cn c quy ch ghi nhn hng ho lu thng trong nc v hng ho xut khu, nhp
khu ca th tng chnh ph
+ Cn c quy ch cng b tiu chun sn phm thc phm ban hnh km theo quyt
nh s 42/2005/Q-BYT ngy 08 Thng 12 nm 2005 ca B trng B Y t. Chng
ti tin hnh xy dng tiu chun c s p dng cho sn phm thc phm chc nng
vin nang mm bo ng Haliotis vi cc ch tiu c th c trnh by trong bo co
chnh.
Qui cch ng gi:
Khi lng tnh: +170 gam/l (1700mg/vin x 100 vin)
+ 17gam/v (1700mg/vin x 10 vin x10 v/hp)
Cng dng:
Bi b sc kho, tng cng sinh lc, gip tng th lc, sng mt v h tr iu
tr u chong, au nhc xng khp.
Liu dng: Dng 2 vin/ln x 2 3 ln /ngy
Hng dn bo qun, vn chuyn: ni kh ro, thong mt, trnh nh sng mt tri.
- Tin hnh cng b sn phm di dng thc phm chc nng vi B y t xin cp s
ng k lu hnh ton quc:
Sn phm c Cc An ton v sinh thc phm B Y t cp chng nhn tiu
chun sn phm cho vin nang mm bo ng s: 6714/2007/YT-CNTC. Ph hp vi cc
qui nh hin hnh v cht lng, v sinh an ton thc phm v c php lu hnh ton
quc ng nh tiu chun cng b.

Hnh XI.2.1.b. Sn phm vin nang mm bo ng

140

Hnh XI.2.1.b. Chng nhn tiu chun sn phm vin nang mm bo ng

XI.2.2. kt qu Nghin cu c tnh ca vin nang Haliotis
XI.2.2.1. Th c tnh cp :
Sau khi ung nc ct v mu th, chut tt c cc nhm u hot ng, n ung
bnh thng k c nhm ung 3 ln vi th tch ti a cho chut th nghim (0,5 ml/ 10g
chut/ln). Khng c chut no cht trong thi gian theo di.
XI.2.2.2. Th c tnh bn trng din
Tnh trng th: Trong thi gian th nghim, hu ht th c hai nhm hot ng bnh
thng, n ung tt, phn kh, lng mt; khng c hin tng rng lng hoc kh
cng.
Quan st i th cc t chc: Khi kt thc th nghim, tin hnh m th, quan st cc t
chc gan, thn, tim, phi. Khng nhn thy biu hin khc thng trn cc t chc ca
th nhm th so vi nhm chng.
Quan st m bnh hc trn tiu bn vi th: Phn tch m bnh hc c thc hin ti
Khoa Nghin cu thc nghim - Vin Y hc c truyn Qun i
Hnh thi vi th gan sau ung vin nang Haliotis
- L chng: cu trc ca gan bnh thng.
- L th nghim: cc b gan, t bo khng hoi t. Cc xoang mch khng gin,
khng xung huyt.
Hnh thi vi th thn:
- L chng: Cc cu thn khng xung huyt
- L th nghim: cu thn v ng thn bnh thng, khng thy thoi ho biu m ng
thn
141

XI.2.3. nh gi tc dng bi b c th ca vin nang bo ng

XI.2.3.1. Ni dung nghin cu
- Nghin cu tc dng bi b sc kho, tng cng sinh lc, gip tng th lc, sng mt
v h tr iu tr u chong, au nhc xng khp ca vin nang Haliotis trn lm sng.
- Theo di tc dng khng mong mun ca vin nang mm Haliotis.
XI.2.3.2. nh gi hiu qu iu tr chng h lao sau 4 tun
a. S ci thin cc triu chng lm sng trc v sau thc nghim
Sau 4 tun thc nghim ta thy cc triu chng lm sng nh mt mi gim t 30
cn 8 bnh nhn, au u gim t 21 cn 10 bnh nhn chng mt, hoa mt gim t 18
cn 9 bnh nhn, hay qun gim t 16 cn 7 bnh nhn, ng tm phin nhit gim t 9
cn 3 bnh nhn, au lng mi gi gim t 14 cn 4 bnh nhn.
b. S thay i im trung bnh Crocq - Bugard trc v sau thc nghim
S im trung bnh Crocp - Bugard trc thc nghim l 39,73, sau 4 tun thc
nghim l 26,21 (gim 34%); s im gim c ngha thng k vi p < 0,001.
c. S thay i im trung bnh Wechsler trc v sau thc nghim
S im trung bnh Wechsler trc thc nghim l 48,49, sau 4 tun thc nghim
l 63,16 (tng 30%); s im tng c ngha thng k vi p < 0,001.
d. S thay i cn nng trung bnh trc v sau thc nghim
Cn nng trung bnh trc thc nghim l 42,95 kg, sau 4 tun thc nghim l
43,35 kg, s thay i cn nng trung bnh c ngha thng k vi p < 0,05.
e. S thay i c lc trung bnh trc v sau thc nghim
C lc trung bnh tay phi trc thc nghim l 18,45 kg,sau4 tun iu l 19,75
kg (tng 7%); s tng c lc trung bnh c ngha thng k vi p < 0,05.
C lc trung bnh tay tri trc thc nghim l 17,14 kg, sau 4 tun thc nghim
l 18,35 kg(tng 7,5% ); s tng c lc trung bnh c ngha thng k vi p < 0,05.
f. S ci thin v n trc v sau thc nghim
Sau 4 tun thc nghim s bnh nhn n tt tng t 2 bnh nhn ln 18, c
ngha thng k vi p < 0,05, s bnh nhn n km gim t 25 bnh nhn xung cn 10,
c ngha thng k vi p < 0,05.
g. S ci thin v ng trc v sau thc nghim
Sau 4 tun thc nghim s bnh nhn ng tt tng t 2 bnh nhn ln 18, c
ngha thng k vi p < 0,05, s bnh nhn ng km gim t 21 bnh nhn xung cn 5,
c ngha thng k vi p < 0,05.
XI.2.3.3. Tc dng ca vin nang Haliotis trn chc nng gan, thn, nhm m mu
v huyt hc
a. nh gi tc dng ca vin nang Haliotis trn chc nng gan, thn, nhm m
mu
Kt qu thu c cho thy sau 4 tun thc nghim vin nang Haliotis c tc
dng lm lng TC, TG, LDL-C gim v lm tng lng HDL-C trong mu, c ngha
thng k vi p < 0,05.
Bng XI.2.3.a So snh phn tch tnh hnh chc nng gan trc v sau thc nghim
( x s)
Nhm
n=45
p

Trc T
Sau T

AST
42,464,97
31,242,23
>0,05

ALT
28,542,73
27,131,34
>0,05
142

GGT
39,764,67
34,681,65
>0,05

Bng XI.2.3.b. So snh phn tch tnh hnh chc nng thn trc v sau thc nghim
( x s)
n=45

Nhm
Trc T
Sau T

URE
5,091,07
5,160,68
>0,05

CREA
101,758,42
87,522,51
>0,05

Qua bng XI.2.3.a v XI.2.3.b, so snh gi tr trung bnh ca chc nng gan v thn
trc v sau thc nghim cho thy khng c thay i, khng c ngha thng k (p > 0,05).
b. S thay i cc ch s huyt hc trc v sau thc nghim
Bng 3.3.3.d. S thay i cc ch s huyt hc trc v sau thc nghim ( x s)
Thi gian
D0 ( x s)
D60 ( x s)
p
S lng hng cu (M/ul)
Hemoglobin (g/dl)
S lng bch cu (K/ul)
Protit mu ton phn (g/l)

4,11 0,12
12,34 0,32
7,40 0,45
69,271,51

4,62 0,05
13,130 0,28
8,05 0,37
71,84 1,36

< 0,001
< 0,05
> 0,05
< 0,05

Sau 4 tun thc nghim nhn thy s tng hng cu trong mu ngoi vi c
ngha thng k (p < 0,001), tng Hb trong mu ngoi vi c ngha thng k (p < 0,05),
tng protit mu c ngha thng k (p < 0,05). So snh gi tr trung bnh trc v sau
thc nghim thy s thay i bch cu khng c ngha thng k (p > 0,05).
XI.2.3.4. Theo di tc dng khng mong mun ca vin nang Haliotis trn lm
sng: Trong qu trnh thc nghim tt c cc bnh nhn u khng thy xut hin cc
triu chng bun nn, a chy, y bng, mn nga.
XI.2.3.5. Nhn xt v bn lun
Sau 4 tun thc nghim ta c th d nhn thy cc triu chng lm sng c ci
thin r rt: Mt mi gim t 30 cn 8 bnh nhn, au u gim t 21 cn 10 bnh nhn
chng mt, hoa mt gim t 18 cn 9 bnh nhn, hay qun gim t 16 cn 7 bnh nhn,
ng tm phin nhit gim t 9 cn 3 bnh nhn, au lng mi gi gim t 14 cn 4 bnh
nhn. Mc suy nhc, ri lon tr nh, n v ng c ci thin r rt. Th trng,
trng lc c u tng sau 4 tun thc nghim.
Khng c s bin i chc nng gan v thn. Ngoi ra qua qu trnh thc nghim
thy lng m trong mu gim, tng s lng hng cu, hemoglobin v protit mu ton
phn. Khng thy xut hin cc triu chng bun nn, a chy, y bng, mn nga.
XI.4. Kt lun
Vin nang Haliotis bo ch t Bo ng: c cng b lu hnh sn phm di
dng thc phm chc nng, c B Y t cp s ng k lu hnh ton Quc. Qua
nghin cu c tnh v nh gi tc dng trn lm sng cho thy:
1. V c tnh:
- Khng xc nh c liu gy cht 50% ng vt th nghim (LD50) v khng tm
c liu gy cht chut mc d cho ung vi mc liu cao.
- Cho th ung lin tc trong mt thng vi mc liu 2 vin/kg/ngy khng nhn thy
s khc nhau ng k v tnh trng th, cc ch s huyt hc v sinh ho, i th cc
tng ph sau th nghim gia nhm chng v nhm th; hnh nh tn thng m bnh
hc t chc gan v thn trn tiu bn vi th khng c s khc nhau nhiu gia nhm
chng v nhm th.
2. V tc dng bi b c th trn lm sng:
nh gi tc dng h tr iu tr bnh suy nhc c th ca vin nang mm bo
ng trn lm sng sau 4 tun thc nghim ta thy vin nang c tc dng: tng cng
sinh lc, gip tng th lc, sng mt v h tr iu tr u chong, au nhc xng khp.
143

Chng XII. Nghin cu cng ngh sn xut vin nang mm

hi sm Hasamin
XII.1. M u
Phn ny trnh by cc c s khoa hc tin hnh nghin cu sn xut sn phm
chc nng Hasamin t hi sm v c chia thnh cc mc nh sau:
XII.1.1. Hi sm trong kinh nghim dn gian
Mc ny gii thiu s b v hi sm v cng dng ca chng, mt s mn n c
tc dng bi b c th v bi thuc c s dng hi sm trong ng Y. V mt dc liu,
do rt giu dinh dng v tc dng khng km v nhn sm, nn ngi ta coi tht hi
sm l nhn sm ca bin. N cha tnh trng c th gy yu, thn kinh suy nhc,
vim ph qun, cc chng chy mu, ho, mn nht. Dng dng thng thng l nng
gin, tn thnh bt.
XII.1.2. Ngun nguyn liu hi sm
Phn ny gii thiu s b v ngun cung cp lng ln mu hi sm, do c th
nui trng vi nng sut cao nn ngun nguyn liu c m bo.
XII.1.3. Nghin cu thnh phn sinh ha mt s loi hi sm
Mc ny trnh by cc nghin cu xc nh thnh phn sinh ha trc y ca cc
loi hi sm: hi sm en: Holothuria vagabunda, hi sm trng: H. scabra, hi sm :
Holothuria sp., hi sm nu: Actynopyga echinites, hi sm xanh: Bohadschia
graeffei, hi sm vng, hi sm da trn: Bohadschia argus (Jaeger) v hi sm v:
Microthele nobilis trong khun kh ti cp nh nc KC09.15 (2003-2005).
Cc kt qu nghin cu cho thy, cc loi hi sm c nghin cu u cha hm
lng cao cc cht dinh dng nh protein, lipit, cc axit amin c bit l cc axit amin
khng thay th, cc axit bo, cc nguyn t vi lng v cc hoocmon. Qua cc kt qu
nghin cu v thnh phn v hm lng cc axt amin, cc nguyn t vi lng c th
gii thch ti sao con ngi li dng hi sm lm thc phm b dng c th v hi sm
cha tp hp cht c tc dng b mu, tng cng trao i cht b thn kinh, b m,
trng dng, tr ha c th. Ngoi ra, vi hm lng cao cc hoocmon c bit l
testosteron, gip ta c c s khoa hc hiu c tc dng ca hi sm i vi kh nng
sinh dc ca nam gii v tng cng sc khe cho ngi gi.
XII.1.4. Nghin cu thnh phn cc cht c hot tnh sinh hc
ca mt s loi hi sm
Mc ny trnh by cc nghin cu trc y v thnh phn hot cht ca mt s
loi hi sm nh: Holothuria vagabunda, H. martensii... Kt qu ng ch l chng
ti phn lp v xc nh c t c hai hoi hi sm ny 02 hp cht tritecpen
saponin dng khung holosthan l: holothurin A va holothurin B vi hm lng kh cao.
Chng l cc hp cht c cu trc ha hc rt c o v th hin nhiu hot tnh sinh
hc ng quan tm. Cc nghin cu nh gi hot tnh sinh hc cho thy: cc hp cht
ny u th hin hot tnh gy c t bo cao i vi nhiu dng t bo ung th ngi
nh KB (ung th biu m), FL (ung th mng t cung), Hep-G2 (ung th gan). Ngoi ra,
hp cht holothurin A cn th hin hot tnh khng MAO mnh (IC50= 1,5 g/ml).
XII.1.5. C s hnh thnh sn phm
T cc kinh nghim c truyn cng vi s minh chng bng cc nghin cu hin
i v thnh phn ha hc v hot tnh sinh hc cho thy, hi sm l loi ng vt c
gi tr dinh dng rt cao vi hm lng di do ca cc loi protein, lipit, cc axit bo,
cc axit amin khng thay th, cc loi vitamin, khong cht, cc nguyn t vi lng....
Mc d cc nguyn t vi lng tn ti trong c th vi khi lng v cng nh nhng n
144

ng vai tr quyt nh trong hu ht cc phn ng sinh ha v cc qu trnh trao i

cht trong c th. Vi hm lng cc nguyn t vi lng kh y v di do trong hi
sm lm cho n tr thnh mt loi thc phm c gi tr rt cao.
Hn th na hi sm cn cha nhiu loi hoocmon quan trng, c bit l
testosteron vi hm lng (20,143 ng/g) nhiu gp 3 ln trong yn so (8,45 ng/g), mt
hooc mn c bit n c vai tr quan trng trong vic pht trin cc du hiu sinh
dc ph, to ra dc tnh nam gii. Testosteron nh hng n qu trnh chuyn ha ca
nit v photpho trong c th, n c tc dng kch thch qu trnh sinh tng hp protein,
gi nit, lm gim bi tit ur, lm pht trin cc c.
Cc nghin cu v thnh phn ha hc cho thy hi sm cha rt nhiu cht
c cc hot tnh qu bu nh hot tnh gy c t bo, chng xy ha, khng MAO. c
bit l s c mt ca cc hp cht holothurin c hot tnh gy c t bo rt mnh i
vi nhiu dng t bo ung th ngi, ha hn cho vic pht trin cc dc phm c tc
dng phng v tr bnh ung th. Nh vy hi sm khng nhng c gi tr dinh dng rt
cao m cn c tc dng phng v cha bnh.
T cc nghin cu ca chng ti, cng vi vic ngun nguyn liu hi sm c th
c ch ng m bo bng cc phng php nui trng, t c s khoa hc vng
chc cho vic xy dng, sn xut v pht trin mt loi thc phm chc nng c gi tr
cao l Vin nang mm hi sm: HASAMIN. iu ny s gp phn nng cao tnh n
nh v gi tr dinh dng cng nh gi tr dc liu ca hi sm, gp phn s dng
ngun nguyn liu qu gi ny mt cch hp l v c hiu qu nht .
XII.2. kt qu v tho lun
XII.2.1. xy dng quy trnh cng ngh sn xut vin nang mm
hi sm Hasamin
Phn ny thuyt minh chi tit cc cng on trong quy trnh cng ngh sn xut
vin nang mm hi sm Hasamin.
- Tri qua nhiu cng on sn xut khc nhau, mi vin nang mm hi sm Hasamin
thnh phm c cha:
200 mg
- Bt hi sm
170 mg
- Du u nnh
120 mg
- Du c
190 mg
- Sp ong trng
90 mg
- Gelatin
30 mg
- Glycerin
10 mg
- Dung dch Sorbitol 70%
0,1 mg
- Nipagin
0,03 mg
- Nipasol
1 mg
- Etyl vanilin
10 mg
- Titan dioxit
1 mg
- Mu Brown R
- Ch nh
Tng cng sinh lc, duy tr trng thi hot ng cao, chng stress, chng lo ha,
tng cng min dch, tng sc khng ca c th, b sung axit amin, cc nguyn t vi
lng, tng tun hon mu, ci thin kh nng hp th xy, chng mi c tim, thc y
qu trnh chuyn ha v hp th, tng sinh tng hp protein, gim cholesterol mu.
- Cch dng v liu dng:
Ung mi ln 2 vin, ngy 2 ln.
- Bo qun, hn dng:
Bo qun ni kh ro, nhit 15-25oC,
- hn dng:
24 thng k t ngy sn xut
145

Quy trnh cng ngh sn xut vin nang mm hi sm Hasamin c tm tt trn

s XII.2.1 sau:
Du u nnh,
du c

Bt hi sm,
Gelatin, glycerin,
Sp ong trng dd Sorbitol, nc ct

Cn

Cn

Hn hp 1

Hn hp 2

Hn hp thuc

Cht mu,
titan dioxyt,
nc ct

Ethyl vanilin,
nc ct

Cht bo
qun, etanol
90%

Cn

Cn

Cn

Cn

Kim nghim
bn thnh
phm

Hn hp v nang

ng nang

Sy nang

Chn nang

p v

ng gi
Kim nghim
thnh phm
Thnh phm,
Nhp kho

S XII.2.1. Quy trnh cng ngh sn xut vin nang mm hi sm HASAMIN

146

XII.2.2. Nghin cu xc nh Thnh phn, th nghim c tnh cp,

c tnh bn trng din v tc dng trn ng vt ca sn
phm vin nang mm hi sm Hasamin
XII.2.2.1. Xc nh thnh phn vin nang mm HASAMIN
a. Xc nh thnh phn dinh dng, ch tiu ha l, vi sinh ca sn phm
Sau khi xc nh c cc iu kin cng ngh ca quy trnh sn xut vin
nang mm hi sm HASAMIN, vic sn xut c tin hnh. Sn phm to ra c
phn tch cc ch tiu cht lng v kt qu u nm trong gii hn cho php.
Sn phm c xy dng tiu chun c s v c B Y t cp chng nhn
tiu chun sn phm s 4978/2006/YT-CNTC vi tn gi : Thc phm chc nng Vin
nang mm hi sm HASAMIN.
- Trng thi: dng vin nang mm
- Mu sc: mu nu en
- Mi: mi thm, hi tanh c trng ca sn phm
- V: c v mn
- tan r: khng qu 30 pht
- ng u khi lng: 7,5% so vi khi lng trung bnh thuc trong nang
b. Xc nh thnh phn axit amin trong vin Hasamin
Qua kt qu nghin cu cho thy hm lng cc axit amin c trong sn phm vin
nang Hasamin c thnh phn axit amin tng i cao khng km g so vi cc sn phm
khc trong nc v ngoi nc nh ru Hi m, vin Taxaton, vin Triplex hay gi
khang thai ca Trung Quc. sn phm gi Khang thai, thnh phn c 18 axit amin v
hm lng cng tng i cao nh: Histidin: 9,88; Xistein: 9,54; Lxin: 8,21;
A.Aspactic: 7,22, Trong khi vin tng lc Hasamin thnh phn cng c ti 17 axit
amin v hm lng cng rt phong ph nh: A.Aspactic: 2,87; A.glutamic: 3,97;
Treonin: 2,96; Xerin: 3,09; Glyxin: 5,98; Tirozin: 3,12; Alanin: 3,26; Acginin: 6,24;
Histidin: 2,98; Valin: 3,10; Metionin: 3,12; PhenylAlanin: 4,06; Izolxin: 5,12; Lxin:
4,80; Lizin: 1,12; 4-Hydroprolin: 4,38; Prolin: 0,98.
Vi thnh phn axit amin ca vin tng lc HASAMIN cho thy gi tr chc
nng dinh dng khng km g so vi cc sn phm tng t trong nc v nc
ngoi, do vy sn phm Hasamin dng cho vic nng cao th lc ca vn ng vin l
rt tt.

Hnh XII.2.2.1.a. Sn phm nang mm hi Sm HASAMIN

147

Hnh XII.2.2.1.b. Chng nhn tiu chun sn phm vin nang mm

hi sm HASAMIN
c. Nghin cu thnh phn cc cht vi khong trong vin Hasamin
V hm lng cc hot cht vi khong c trong vin Hasamin qua kt qu nghin
cu cho thy: Hm lng ng (Cu) hu c c trong vin Hasamin l 3,73 mg/kg vck,
St (Fe) l 19,89 mg/kg vck, km (Zn) hu c l 8,20, hm lng km l 8,20 mg/kg vck
v Mangan (Mn) l 37,19mg/kg vck. Nhn chung hm lng cc cht vi khong c trong
vin tng lc Hasamin so vi cc sn phm tng t trong nc cng nh so vi sn
phm gi khang thai v vin Triplex ca Trung Quc th kt qu cng khng khc nhau
nhiu. Thm ch mt s thnh phn ca hm lng nguyn t km hay mangan (Mn)
cn cao bng hoc hn.
d. Nghin cu thnh phn cc hot cht hoocmn steroit trong vin Hasamin
Kt qu phn tch hm lng cc hot cht steroit c trong vin HASAMIN l
kh cao so vi cc sn phm khc, k c sn phm ngoi.
Hm lng Testosteron (mg/kg vck) c trong vin HASAMIN l (62,17), v
ru Hi m l (11,02), vin Taxaton (68,00) v gi khang thai ca Trung quc (67,35)
nhng cao hn vin Triplex ca Trung quc (32,08).
Hm lng Progesteron (mg/kg vck) ca vin HASAMIN l (65,24) cao hn
nhiu so vi cc sn phm trong nc v ca Trung Quc nh ru Hi m (9,28), vin
Taxaton (65,07), gi khang thai (52,15) v vin Triplex (38,72).
Hm lng Cortisol (mg/kg vck) ca vin HASAMIN l (17,53) cao hn tt c
cc sn phm trong nc nh ca ru Hi m (8,22), vin Taxaton (19,40) v khng
thua km g cc sn phm ca Trung Quc nh gi Khang Thai (59,24), vin Triplex
(55,12).

148

XII.2.2.2. Nghin cu c tnh cp v c tnh bn trng din ca vin nang mm

hi sm HASAMIN
c tnh cp v c tnh bn trng din c tin hnh nh gi ti Vin Y hc
c truyn Qun i v thu c cc kt qu sau:
a. c tnh cp:
Bng XII.2.2.2.a: B tr th nghim v kt qu th c tnh cp:
Nhm
1
2
3
4
5
6

Liu dng
(ml/ 10g chut)
0,2 ml x 1 ln
0,2 ml x 3 ln
0,3 ml x 3 ln
0,4 ml x 3 ln
0,5 ml x 3 ln
0,5 ml nc x 3 ln

Liu dng
(g / kg chut)
2,0 g
6,0 g
9,0 g
12,0 g
15,0 g
nc ct

S chut th
nghim
10
10
10
10
10
10

S chut cht
0
0
0
0
0
0

Nhn xt: Sau khi ung nc ct v mu th, chut tt c cc nhm u hot ng, n
ung bnh thng k c nhm ung 3 ln vi th tch ti a cho chut th nghim (0,5
ml/ 10g chut/ln). Khng c chut no cht trong thi gian theo di.
b. c tnh bn trng din
- Tnh trng th: Trong thi gian th nghim, hu ht th c hai nhm hot
ng bnh thng, n ung tt, phn kh, lng mt; khng c hin tng rng lng
hoc kh cng.
- Quan st i th cc t chc: Khi kt thc th nghim, tin hnh m th, quan
st cc t chc gan, thn, tim, phi. Khng nhn thy biu hin khc thng trn cc t
chc ca th nhm th so vi nhm chng.
- Quan st m bnh hc trn tiu bn vi th: Phn tch m bnh hc c thc
hin ti Khoa Nghin cu thc nghim - Vin Y hc c truyn Qun i.
Hnh thi vi th gan sau ung thuc:
+ L chng: cu trc ca gan bnh thng.
+ L th nghim: cc b gan, t bo khng hoi t. Cc xoang mch khng gin,
khng xung huyt.
Hnh thi vi th thn:
+ L chng: Cc cu thn khng xung huyt
+ L th nghim: cu thn v ng thn bnh thng, khng thy thoi ho biu m
ng thn
c. Kt lun
Mu vin nang HASAMIN, l SX: 110707 gi ti yu cu th c tnh cp v bn
trng din c cc kt qu nh sau:
- Th c tnh cp trn chut nht trng: cho ung hn dch mu th trong nc t
1-3 ln cch nhau 3 gi trong ngy, vi tng liu t 2,0 15,0g/ kg khng nhn thy
biu hin ng c no trn chut th nghim trong thi gian theo di (k c nhm chut
ung vi liu ti a 0,3g/ 10g chut/ ln). Khng xc nh c liu gy cht 50% ng
vt th nghim (LD50) v khng tm c liu gy cht chut mc d cho ung vi
mc liu cao.
- Th c tnh bn trng din: cho th ung lin tc trong mt thng vi mc liu 2
vin/kg/ ngy c cc kt qu nh nu trn. Cha nhn thy s khc nhau ng k v
tnh trng th, cc ch s huyt hc v sinh ho, i th cc tng ph sau th nghim gia
nhm chng v nhm th. Hnh nh tn thng m bnh hc t chc gan v thn trn
tiu bn vi th khng c s khc nhau nhiu gia nhm chng v nhm th.

149

XII.2.2.3. Th nghim tc dng ca vin Hasamin trn ng vt thc nghim

Kt qu cho thy: b sung vin Vin Hasamin c tc dng lm tng c ngha v
phm cht tinh dch th so vi b sung Placebo.
Trc khi b sung ch phm, lng tinh dch th khc nhau khng c ngha gia
cc l th nghim. Sau 10 ngy b sung, lng tinh dch ca l 1: 0,78ml cao hn c
ngha so vi lng tinh dch ca l 2: 0,56ml v trc khi b sung. Sau 20 ngy b sung
ch phm, lng tinh dch ca cc l cao hn khng c ngha so vi sau 10 ngy.
Lng tinh dch ca l1 cao hn c ngha ngha so vi l 2.
Trc khi b sung ch phm, hot lc tin thng ca tinh trng khc nhau khng
c ngha gia cc l th nghim. Sau 10 ngy b sung ch phm, c tc dng tt lm
tng hot lc tin thng ca tinh trng l 1. Sau 20 ngy, hot lc tin thng ca tinh
trng l1: 74,70% cao hn c ngha so vi l2: 62,73%. Hot lc tin thng ca tinh
trng l 1 sau khi b sung ch phm cao hn c ngha so vi trc khi b sung.
Nng tinh trng khc nhau khng c ngha trc khi b sung ch phm. B
sung ch phm c tc dng tng c ngha v nng tinh trng l 1 so vi l 2 v
trc khi b sung.
B sung ch phm c tc dng tng tng s tinh trng tin thng. Sau 20 ngy b
sung, tng s tinh trng c hot lc tin thng l 1:181,87 triu, cao hn c ngha so
vi l 2: 89,25 triu/ln.
T l k hnh ca tinh trng trc khi b sung ch phm khc nhau khng c
ngha. Sau 10 ngy b sung ch phm, t l k hnh ca tinh trng l 1: 21,74% thp
hn c ngha thng k so vi l 2: 25,00%.
T l sng ca tinh trng khc nhau khng c ngha trc khi b sung ch phm.
Ch phm c nh hng tt lm tng t l sng ca tinh trng. Sau 20 ngy b sung
Hasamin t l sng ca tinh trng l 1: 80,72% cao hn c ngha so vi l 2:
69,83%.
Ch phm c nh hng tng sc khng ca tinh trng l 1 so vi trc b sung,
nhng khng c ngha.
Ch phm duy tr c pH n nh.
XII.3. Kt lun.
# Sn phm vin Hasamin phn tch cho thy hm lng qua thc nghim trn ng
vt cho thy an ton v c tc dng rt tt. Vin Hasamin l nhng ngun thc phm
chc nng c gi tr dinh dng. Th hin cc hm lng axit amin, hoocmn
steroit v cc nguyn t vi lng l cao so vi mt s loi sn phm tng t khc
trong nc v nc ngoi.
# Cc th nghim v c tnh cp v c tnh bn trng din trn chut nht trng v
th cho thy: khng nhn thy biu hin ng c no trn chut th nghim trong
thi gian theo di, Khng xc nh c liu gy cht 50% ng vt th nghim
(LD50) v khng tm c liu gy cht chut mc d cho ung vi mc liu cao,
khng nhn thy s khc nhau ng k v tnh trng th, cc ch s huyt hc v
sinh ho, i th cc tng ph sau th nghim gia nhm chng v nhm th. Hnh
nh tn thng m bnh hc t chc gan v thn trn tiu bn vi th khng c s
khc nhau nhiu gia nhm chng v nhm th.
# Vin Hasamin c tc dng thc s r nt thng qua vic thc nghim tng cht
lng tinh dch trn th.

150

phn III. Kt lun v kin ngh chung

A. Kt lun
ti KC09.09/06-10 Nghin cu sng lc cc cht c hot tnh sinh hc theo
nh hng khng sinh, gy c t bo v chng xy ho t sinh vt bin nhm to cc sn
phm c gi tr dc dng l ti nghin cu lin ngnh, t vn nghin cu mt

cch c h thng ngun dc liu bin Vit Nam p dng m hnh nghin cu ho hc
theo nh hng hot tnh sinh hc (bioasay-guided chemical study). Nh chng ti
trnh by trn, y l m hnh nghin cu tin tin nht hin nay trn th gii, c
nhiu quc gia nh M, Nht Bn, Hn Quc, Php p dng hnh thnh nhng
chng trnh nghin cu ln qui m quc gia v xuyn quc gia. Kt qu l trong nhng
nm va qua, nhiu loi thuc c hiu cha tr cc bnh him ngho c ra i v
lu hnh trn th gii thng qua cc chng trnh ny. Ngay cc nc trong khu vc nh
Thi Lan, Malaysia, Philippines cng c nhng chng trnh sng lc cc cht c
hat tnh sinh hc t bin v t c nhiu kt qu ban u ng khch l. Hiu qu
ca nhng chng trnh sng lc ny ph thuc rt nhiu vo i ng cn b cc chuyn
ngnh lin quan, h thng trang thit b phn tch v c bit l h thng thit b th hot
tnh sinh hc hin i, c tin cy cao. Mc thnh cng cng ph thuc rt nhiu
vo kinh ph dnh cho nghin cu.
trin khai thc hin ti ny, Ban Ch nhim ti KC09.09/06-10 tp
trung c i ng cn b khoa hc ng o thuc cc lnh vc chuyn mn khc nhau
nh Ho, Sinh, Y, Dc, Cng ngh. Cc c quan tham gia ti u l nhng c quan
u ngnh ca Vit Nam cng tham gia gii quyt nhim v. Sau 3 nm trin khai thc
hin, vi n lc c gng ca tp th cn b thc hin ti, c s quan tm, ng vin
rt ln v kp thi ca Ban Ch nhim chng trnh KC09/06-10, ca B Khoa hc v
Cng ngh, Vin Khoa hc v Cng ngh Vit nam, ti hon thnh tt cc mc
tiu ra ban u v t c nhng kt qu tt. Cn c vo cc kt qu t c,
c th rt ra cc kt lun sau:
I. t chc 9 t kho st thu thp mu sinh vt bin trn cc vng bin bc, trung v
nam Vit Nam, thu thp v xc nh tn phn loi 101 mu sinh vt bin v to tiu bn
lu tr lu di tp trung vo cc nhm san h mm, da gai v hi min. xy dng
c c s d liu qun l cc loi sinh vt bin cha dc liu di ven b Vit Nam. C
s d liu cho php qun l, truy cp, b sung, tu chnh thun tin v n gin.
II. tin hnh to dch chit tng v cc dch chit phn on ca 101 mu thu c
theo mt qui trnh thng nht phc v cho sng lc hot tnh sinh hc v nghin cu ha
hc.
III. tin hnh sng lc hot tnh sinh hc ca tt c cc dch chit tng v cc dch
chit phn on ca 101 mu sinh vt bin theo cc nh hng:
a. Hot tnh khng vi sinh vt kim nh (Antimicrobial activity asay)
Kt qu thu c cho thy, s mu c hot hot tnh khng vi sinh vt kim nh
l 51 trong tng s 101 mu sng lc (chim t l 50,49 %). Trong , s mu c hot
tnh khng 1 vi sinh vt kim nh l 8 mu (chim 7,92 %), khng 2 vi sinh kim nh
l 10 mu, khng 3 vi sinh kim nh l 19 mu, khng 4 vi sinh kim nh l 10 mu,
151

khng 5 vi sinh kim nh l 2 mu, khng 6 vi sinh kim nh 1 mu v khng 7 vi sinh

kim nh l 1 mu. Cc mu c hot tnh c tin hnh chit phn b gia nc v
cc dung mi hu c c phn cc tng dn t hexan sang butanol. xc nh gi tr
c ch ti thiu (MIC) ca dch chit MeOH tng v cc phn on chit phn b.
b. Hot tnh gy c t bo trn 3 dng t bo ung th ngi (Cytotoxic activity assay)
Trong s 101 mu th nghim, c 43 mu c biu hin hot tnh gy c t bo
(chim 42,57 %), trong c 12 mu khng mt dng t bo (chim 11,88 %), 15 mu
khng 2 dng t bo (chim 14,85 %) v 16 mu khng c 3 dng t bo (chim 15,84
%). xc nh gi tr IC50 (nng c ch 50% s pht trin ca t bo) ca dch chit
MeOH tng v cc phn on chit phn b nh hng cho cc nghin cu tip theo.
c. Hot tnh chng oxy ho theo c ch bao vy gc t do DPPH (Antioxidant activity
assay)
Kt qu thu c cho thy, c 23 mu trong tng s 101 mu (chim t l 22,72
%) c biu hin dng tnh vi gi tr SC > 50%. Trong , c 18 mu c c kh nng
bao vy t 50-70% cc gc t do trong th nghim DPPH v 5 mu c kh nng bao gy
trn 70% cc gc t do. nh hng cho cc nghin cu tip theo, gi tr SC50 (gi tr
nng mu th c kh nng bao vy 50% cc gc t do) ca dch chit MeOH tng v
cc phn on chit phn b ca cc mu c biu hin hot tnh c xc nh.
IV. Trn c s cc kt qu nghin cu thu thp, gim nh loi, to dch chit v sng
lc hot tnh sinh hc, kt hp vi cc tiu ch la chn b sung nh: Mc kh khn
trong cng tc ly mu, tr lng thc t ca loi v kh nng c th thu thp mu, tra
cu cc nghin cu v hot tnh sinh hc v thnh phn ha hc cng b v loi hoc
chi ca i tng nghin cu..., la chn c 07 loi sinh vt bin thuc cc nhm
san h mm, da gai v hi min thu mu lng ln v nghin cu su v mt ho hc
l :
- Nhm san h mm :
1. T loi san h mm Sarcophyton mililatensis, chit xut, phn lp v xc nh cu
trc c 10 hp cht trong c 07 hp cht ditecpen dng cembranoit (05 cht
mi), 01 hp cht 9,11-secosterol mi (sarcomilasterol) v 02 hp cht sterol mang
nhiu nhm hydroxyl. tin hnh nh gi hot tnh gy c t bo, chng xy ha
v chng long xng ca cc hp cht phn lp c. Cc kt qu c ng ti
trn 02 bi bo chuyn ngnh quc gia v 02 bi bo chuyn ngnh quc t.
2. T loi san h mm Cladiella sp., 08 hp cht c phn lp v xc nh cu trc,
trong c: 02 hp cht 9,11-secosterol (01 cht mi l : cladiasterol), 01 hp cht
ceramit mi (cladiaceramide) v 01 hp cht cerebrosit mi (cladiacerebroside).
tin hnh nh gi hot tnh gy c t bo, chng xy ha v khng NF-B ca cc
hp cht phn lp c. ng ti kt qu trn 01 tp ch chuyn ngnh quc t.
- Nhm da gai:
3. T loi hi sm Holothuria scabra, tin hnh chit xut, phn lp v xc nh c
cu trc ca 09 hp cht, trong c 06 hp cht tritecpen saponin dng khung
holosthan (03 hp cht mi l: holothurin A3, holothurin A4 v holothrigenin B).
tin hnh nh gi hot tnh gy c t bo v khng sinh ca cc hp cht. Tt c cc
hp cht saponin u th hin hot tnh gy c t bo cao. Cc hp cht holothurin
A2 v A3 c nh gi hot tnh gy c t bo bng phng php quan st trc
tip hnh thi b mt t bo di knh hin vi sau 24, 48 v 72 gi. Cc kt qu
c ng trn 02 tp ch chuyn ngnh quc t v 01 hi ngh quc gia. Ngoi ra, tp
th tc gi cng ng k v c cp 01 bng c quyn sng ch s 6852 ti Cc
152

S hu Tr tu Vit Nam v phng php phn lp v hot tnh chng ung th ca hp

cht mi holothurin A3. Cc kt qu nghin cu c gi tham gia gii thng
khoa hc k thut thanh nin ln th XVIII - 2008 v ot gii khoa hc: s
1890/GTKHKTTN-2008.
4. T loi sao bin Archaster typicus, 07 hp cht c phn lp v xc nh cu trc,
trong c 01 hp cht cerebrosit mi (archastercerebroside) v 03 hp cht sterol
mang nhiu nhm hydroxy (highly oxygenated sterols). Hot tnh gy c t bo v
khng NF-B ca cc hp cht c nh gi. ng c 02 bi bo trn tp
ch quc gia, 01 bi trn Hi tho chuyn ngnh quc gia v 01 bi trn Hi tho
chuyn ngnh quc t.
- Nhm hi min:
5. T loi hi min Xestospongia testudinaria, 10 hp cht c phn lp v xc nh
cu trc, trong ng lu nht l c 02 hp cht axit bo khng no b brm ha c
cu trc mi l: axit 20-bromo-(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic
v 22-bromo-(17E,21E)-docosa-17,21-diene-9,11,19-triynoic. tin hnh nh gi
hot tnh khng vi sinh vt kim nh v gy c t bo ca cc hp cht. Hai hp
cht mi u khng mnh ba chng vi sinh vt kim nh l: trc khun m xanh
Pseudomonas aeruginosa, khng t cu vng Staphylococcus aureus v khng nm
si Fusarium oxysporum vi gi tr IC50 thp l 50 g/ml. Tp th tc gi ng
c 02 bi bo trn tp ch chuyn ngnh quc gia. Ngoi ra, tp th tc gi gi
bn tho v n ng k 01 bng c quyn sng ch (s n 1-2007-01616 SC ngy
8/8/2008) ti Cc S hu Tr tu Vit Nam v hp cht axit 20-bromo(11E,15E,19E)-eicosa-11,15,19-triene-7,9,17-triynoic v ang c tin hnh thm
nh ni dung khoa hc.
6. T loi hi min cnh xanh Gellius varius, phn lp v xc nh cu trc c 10
hp cht. Trong c 02 hp cht sulfuno glycolipit c cu trc c th vi s c mt
ca d t lu hunh trong phn t. tin hnh nh gi hot tnh gy c t bo v
khng vi sinh vt kim nh ca cc hp cht. Cc kt qu c ng ti trong 02
bi bo trn cc tp ch chuyn ngnh quc gia.
7. T loi hi min Ianthella sp., 05 hp cht steroit c cu trc vng propan mch
nhnh c phn lp v xc nh cu trc. Trong , 03 hp cht c cu trc mi
v c chng ti t tn l iansterol A, B, v C. Kt qu nghin cu hot tnh gy
c t bo cho thy, tt c cc hp cht phn lp c u th hin hot tnh cao
trn 03 dng t bo ung th ngi c th nghim l MCF-7 (v), SK-Hep-1 (gan)
v Hela (c t cung).
V. nghin cu thm d kh nng khng vim t rong, to bin Vit Nam v thu c
cc kt qu sau:
- thu thp v nh tn c 19 loi rong bin Vit Nam phn b cc tnh min
Bc, Trung v Nam ca Vit Nam.
- xy dng c quy trnh tch dch chit c hot tnh khng vim t cc loi rong
bin Vit Nam bng dung mi metanol; phng php sng lc nhanh cc loi rong to
bin trong khun kh ti c hot tnh khng vim bng dung mi metanol.
- Trn c s ca phng php ny, trong tng s 19 loi rong bin c sng lc th
Sargassum swartzii v Ulva reticulata c hot tnh khng vim cao v c tim nng
khai thc c pht hin.
- tin hnh th hot tnh khng vim v khng sinh ca cc dch chit thu c t
Sargassum swartzii v Ulva reticulata.
- xc nh c hm lng lipit tng s v thnh phn axit bo ca hai mu cao
rong Sargassum swartzii v Ulva reticulata
153

- phn lp v xc nh cu trc c hp cht 3,5-dimethyl-4-al-pyrol t cao

diclometan v hp cht 22-dehydroclerosterol t cao n-hexan ca rong rong
Sargassum swartzii.
VI. nghin cu thm d kh nng khng sinh t vi sinh vt bin v thu c cc kt
qu sau:
- phn lp v lm sch c 1052 chng vi sinh vt bin t 81 mu nc bin, trm
tch bin v ng thc vt bin Vit Nam. Trong , c 552 chng vi khun, 253
chng nm si, 157 chng x khun v 90 chng nm men.
- sng lc s b v th kh nng khng 5 chng VSV kim nh ca 1052 chng vi
sinh vt bin.
- th kh nng dit su hi, gy c t bo ca 142 chng vi sinh vt bin.
- xc nh kh nng sinh cc enzim ngoi bo amylaza, xenlulaza v proteaza ca 82
chng vi sinh vt bin
- xc nh hm lng axit bo khng bo ho mch di ca 6 chng nm men bin.
- tuyn chn c 11 chng vi sinh vt bin c hot tnh sinh hc cao v nh tn
khoa hc 08 chng.
- a ra qui trnh nui cy v tch chit cht khng khun t dch nui cy chng
nm si T10.6
VII. nghin cu hon thin quy trnh cng ngh v trin khai sn xut c 04 sn
phm thc phm chc nng l: thc phm chc nng CEFISH b sung omega-3 ci
thin chc nng chuyn ha lipit mu, huyt p bnh nhn tim mch, vin tng lc
BIONAMINE tng cng th lc cho vn ng vin t sao bin, vin nang mm
HALIOTIS t bo ng c tc dng bi b c th v thc phm chc nng vin nang
mm HASAMIN t hi sm.
VIII. nghin cu c tnh cp v c tnh bn trng din ca cc ch phm trn
ng vt thc nghim. Kt qu cho thy khng sn phm no th hin c c tnh cp v
c tnh bn trng din trn ng vt th nghim. tin hnh cc nghin cu dc l
cc sn phm thc phm chc nng v thu c cc kt qu tt.
IX. xy dng c tiu chun c s cht lng sn phm v c cp giy chng
nhn tiu chun sn phm ca B Y t, cho php lu hnh trn ton quc ca tt c bn
sn phm: sn phm CEFISH c giy chng nhn tiu chun sn phm s
6585/2007/YT/CNTC, sn phm BIONAMINE c giy chng nhn tiu chun sn
phm s 956/2008/YT-CNTC, sn phm HALIOTIS c cp giy chng nhn tiu
chun sn phm s 6714/2007/YT-CNTC v sn phm HASAMIN c cp giy chng
nhn tiu chun sn phm s 4978/2006/YT-CNTC.
X. sn xut lng ca tt c 04 sn phm phc v cho cc nghin cu v c
tnh v tc dng dc l theo h s ng k.
XI. Cc kt qu khc: ng k 02 c quyn sng ch, xut bn c 01 sch
chuyn kho, ng 17 bi bo trong c 11 bi trn cc tp ch chuyn ngnh quc gia
v 06 bi quc t (05 bi ng v 01 bi gi ng), ng 09 bo co ti cc hi
ngh trong c 04 hi ngh quc t v 05 hi ngh trong nc, o to 02 thc s bo
v thnh cng lun n v 02 tin s (01 tin s bo v thnh cng lun n v 01 ang
tin hnh).

154

b. kin ngh
Da trn cc kt qu thu c, tp th cn b thc hin ti xut mt s kin
ngh sau:
1. Cc nghin cu tip theo cn tip tc tp trung vo cc nhm i tng c tim nng
dc dng cao l san h mm, da gai v hi min vi tng hot tnh c th nh : Gy
c t bo, chng long xng, khng sinh
2. Cn m rng quy m sn xut ca cc sn phm phn phi trn c nc, phc v
cho cng tc chm sc sc khe v nng cao cht lng cuc sng ca ngi dn.
3. Tip tc trin khai nghin cu to sn phm t cc ngun nguyn liu di do v c
nhiu trin vng khc nhm to ra cc sn phm c gi tr tng t gp phn nng
cao gi tr ngun dc liu bin, ng thi c cc bin php bo tn v khai thc hp
l ngun ti nguyn v gi ny.
4. Tip tc pht trin nghin cu su hn na, tm kim cc thnh phn c hot tnh
chnh trong cc nguyn liu nghin cu, pht trin thnh cc ch phm thuc cha
bnh phc v cuc sng.
Li cm n
Tp th cn b thc hin ti xin by t lng bit n n
- Ban ch nhim cc chng trnh KH&CN trng im cp nh nc KC 09/0610: Khoa hc v Cng ngh Bin phc v pht trin bn vng kinh t x hi
- Vn phng cc Chng trnh, B Khoa hc v Cng ngh Vit Nam
- Vin Ha hc cc Hp cht Thin Nhin.
ng vin v gip chng ti trong qu trnh thc hin ti.
Nhn dp ny chng ti cng xin chn thnh cm n cc n v cng tham gia
tch cc vi Vin Ho hc cc hp cht thin nhin, cc nh khoa hc trong v ngoi
Vin cng cng tc ti c hon thnh ng theo k hoch.

155

cc cng trnh cng b trong khun kh ti

1. Danh mc cc sng ch
TT Tn tc gi
1 Chu Vn Minh, Phan
Vn Kim, Nguyn Hi
ng, Nguyn Vn
Thanh, L Mai Hng,
Cng Thung, Trn
Thu Hng, Nguyn
Xun Cng.
2 Chu
Vn
Minh,
Nguyn Xun Cng,
Phan Vn Kim, L Mai
Hng, Cng Thung

2. Danh mc cc sch
TT Tn tc gi
1 Chu Vn Minh, Phan
Vn Kim, Nguyn Hi
ng

Tn sng ch
Hp cht Holothurin A3 c hot tnh
chng ung th v phng php chit hp
cht ny t loi hi sm Holothuria
scabra, Bng c quyn sng ch s
6852, cp theo quyt nh s 2394/QSHTT, ngy 14 thng 02 nm 2008 ca
Cc S hu Tr Tu Vit Nam
Hp cht axit 20-bromo-(11E,15E,19E)eicosa-11,15,19-triene-7,9,17-triynoic v
phng php chit hp cht ny t loi hi
min Xestospongia testudinaria, s n 12007-01616 SC ngy 8/8/2007

Ghi ch
c
cp

chp
nhn
n

Tn sch, nh xut bn
Ghi ch
Cc hp cht c hot tnh sinh hc t sinh xut
vt bin. Nh xut bn Khoa hc K bn
thut, 2007, 241 trang.

3. Danh mc cc tp ch quc t
TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Nguyen Xuan Cuong, New Cembranoid Diterpenes from the
Tran Anh Tuan, Phan Vietnamese Soft Coral Sarcophyton
Van Kiem, Chau Van mililatensis
Stimulate
Osteoblastic
Minh, Eun Mi Choi, and Differentiation in MC3T3-E1 Cells.
Young Ho Kim.
Chemical & Pharmaceutical Bulletin,
2008, 56(7), 988-992.
2 Chau
Van
Minh, A new 9,11-secosterol from the
Nguyen Xuan Cuong, Vietnamese Sea soft coral Sarcophyton
Tran Anh Tuan, Eun Mi mililatensis, increases the function of
Choi, Young Ho Kim, osteoblastic MC3T3-E1 cells, Natural
and Phan Van Kiem.
Product Comunications (USA), 2007,
2(11), 1095-1100.
3 Nguyen Hai Dang, Two new triterpene glycosides from the
Nguyen Van Thanh, Vietnamese sea cucumber Holothuria
Phan Van Kiem Le Mai scabra, Archives of Pharmacal Research,
Huong, Chau Van Minh, 2007, 30(11), 1387-1391.
and Young Ho Kim.
4 Chau Van Minh, Phan New 9,11-secosterol glycoside and
Van Kiem, and Nguyen ceramide from a soft coral Cladiella sp.
Xuan Cuong.
living in Vietnamese sea, ASEAN Journal
on Sicience and Technology for
Development, 2007, 24(3), 275-283.
5 Nguyen Van Thanh, A new triterpene glycoside from the sea
Nguyen Hai Dang, Phan cucumber Holothuria scabra collected in
156

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ng

ng

ng

Van Kiem, Nguyen

Xuan Cuong, Hoang
Thanh Huong, and Chau
Van Minh.
Chau
Van
Minh,
Nguyen Huu Tung,
Nguyen Xuan Nhiem,
Phan Van Kiem, Tran
Thu Ha, Jae-Hee Hyun,
Hee-Kyoung
Kang,
Young Ho Kim

Vietnam. ASEAN Journal on Science and

Technology for Development, 2006,
23(4), 253-259.
C29-Sterols with a cyclopropane ring at C- gi
25 and 27 from the Vietnamese marine ng
sponge Ianthella sp.

4. Danh mc cc tp ch quc gia

TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Chau
Van
Minh, Highly oxygenated sterols from the
Nguyen Xuan Cuong, starfish Archaster typicus. Journal of
Nguyen Van Thanh, Chemistry (Vietnam), 2007, 45(3), 377Hoang Thanh Huong, 381.
and Phan Van Kiem.
cholesterol
and
2 Phan
Van
Kiem, Cholest-7-en-3-one,
Nguyen Van Thanh, batilol from the starfish Archaster typicus.
Nguyen Xuan Cuong, Journal of Chemistry (Vietnam), 2007,
and Chau Van Minh.
45(4), 513-517.
3 Nguyn Xun Cng, Cc hp cht glycolipit t loi hi min
Phan Vn Kim, Chu cnh xanh Gellius varius sinh sng ti
Vn Minh.
vng bin Vit Nam, Tp ch Khoa hc v
Cng ngh, 2007, 45 (2), 41-47.
4 Nguyen Xuan Cuong, Sarcophytol X, a new cembranoid
Tran Anh Tuan, Phan diterpene from the vietnamese sea soft
Van Kiem, and Chau coral Sarcophyton mililatensis, Tp ch
Van Minh.
khoa hc (i hc S phm H Ni II),
2007, 1, 55-60.
5 Chau
Van
Minh, Polyhydroxylated sterols from the
Nguyen Xuan Cuong, Vietnamese Sea soft coral Sarcophyton
Tran Anh Tuan, Eun Mi mililatensis, increases the function of
Choi, Young Ho Kim, osteoblastic MC3T3-E1 cells, Tp ch
and Phan Van Kiem.
khoa hc (i hc S phm H Ni II),
2007, 1, 108-116.
6 Nguyn Xun Cng, Nghin cu thnh phn ha hc loi hi
Trn Anh Tun, Nguyn min Xestospongia testudinaria thu thp
Thu Phng, V Vn ti Vit Nam. Tp ch Khoa hc v Cng
Thnh, Phan Vn Kim, ngh, 2007, 45(3), 43-50.
Chu Vn Minh.
7 Nguyen Xuan Cuong, Isoprenoids from the sponge Gellius
Nguyen Hai Dang, Do varius living in Vietnamese sea, Journal
Cong Thung, Tran Thu of Chemistry (Vietnam), 2007, 45(6), 776Huong, Nguyen Thi 780.
Kim Thuy, Phan Van
Kiem, Chau Van Minh.
157

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ng

ng

ng

ng

10

11

Chau
Van
Minh,
Nguyen Hai Dang,
Nguyen Xuan Cuong,
Phan Van Kiem, Hoang
Thanh Huong,
Nguyn Minh Thanh,
Luyn Quc Hi, Li
Kim Dung, Phan Vn
Kim, Chu Vn Minh,
ng Dim Hng
ng
Dim
Hng,
Hong Minh Hin,
Nguyn Minh Thanh,
Chu
Vn
Minh,
Nguyn Trng Thng
Chau Van Minh, Phan
Van Kiem, Tran Anh
Tuan, and Nguyen Xuan
Cuong.

Bioacive constituents from marine ng

organisms inhabiting in Vietnames sea.
Journal of Science and Technology
(Vietnam), 2007, 45(6-DB), 1-18.
Xc nh bn cht ha hc ca cht khng ng
vim c trong dch chit ton phn t
Sargassum swartzii ca Vit Nam, Tp
ch Cng ngh Sinh hc, 2008, S 6(4A),
713-720.
Nghin cu kh nng khng vim t rong ng
to bin Vit Nam, Tp ch Ha hc,
2008, Tp 46, s 53, trang 81-90.
Two new brominated fatty acids from the ng
sponge Xestospongia testudinaria living in
Vietnamese sea. Advances in Natural
Sciences, 2008, 9(2), 167-171

5. Danh mc cc hi ngh quc t

TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
1 Chau
Van
Minh, A new galactocereboside from the starfish
Nguyen Van Thanh, Archaster typicus, 12t Asian Chemical
Nguyen Xuan Cuong, Congress (12ACC), August 23-25, 2007,
Phan
Van
Kiem, Malaysia, p. 309.
Nunziatina De Tommasi
2 Chau
Van
Minh, Two new triterpene glycosides from the
Nguyen Van Thanh, Vietnamese sea cucumber Holothuria
Nguyen Hai Dang, Phan scabra, 12t Asian Chemical Congress
Van Kiem, Le Mai (12ACC), August 23-25, 2007, Malaysia,
p. 309.
Huong, Young Ho Kim
3 Phan Van Kiem, Chau Bioactive Constituents from Marine
Van Minh, Nguyen Hai organisms inhabiting in vietnamese sea,
prsentation
in
Intenational
Dang, Nguyen Xuan Oral
Cuong, Hoang thanh Conference From the new drug
Development to Human Well-being The
huong, Kim Young Ho
Pharmaceutical Society of Korea, April
30 - May3, 2008, Vol. 1A, p.15.
4 Nguyen Minh Thanh, The screening compound of antiHoang Minh Hien, inflamatory activities from Vietnamese
Hoang Sy Nam, Chau seaweeds. Vth Asian Pacific Phycological
Van Minh, Dang Diem Forum, Wellington, New Zealand,
November 10-14, 2008. p.88
Hong.

Ghi ch
ng

ng

ng

ng

6. Danh mc cc hi ngh quc gia

TT Tn tc gi
Tn bi bo, tp ch, nh xut bn
Ghi ch
1 Nguyen Van Thanh, Steroidal constituents from Holothuria ng
Chau Van Minh, scabra living in Vietnamese sea. Tuyn tp
158

Hoang Thanh Huong, cc bi bo khoa hc Hi ngh Khoa hc ln

Phan Van Kiem.
th 20-HBK H Ni, phn ban Cng ngh
Ha hu c, 2006, 200-204.
Chu
Vn
Minh, Mt galactocereboside mi t sao bin
Nguyn Vn Thanh, Archaster typicus. Hi ngh Ha hc hu c
Nguyn Xun Cng, Ton quc ln th 4, H Ni, thng 10 nm
Phan
Vn
Kim, 2007, trang 446-450.
Nunziatina
De
Tommasi, Lm Ngc
Thim
Hong S Nam, inh Bc u nghin cu sng lc cht c hot
Khnh Chi, Hong tnh khng vim t mt s loi rong bin
Minh Hin, Chu Vit Nam, Nhng Vn nghin cu c bn
Vn Minh, ng trong khoa hc s sng, 2007, 773-776.
Dim Hng
Tng Kim Thun, Nghin cu tuyn chn vi sinh vt bin c
ng Th Mai Anh, kh nng i khng VSV kim nh, gy
L Th Thanh Xun, cht cn trng v kh nng ng dn trong
Phm Cng Hot, nng nghip v y dc. Hi ngh Khoa hc Cc bin php phng tr su bnh khng
Chu Vn Minh
gy nhim mi sinh, 2007, 77-83.
Tng Kim Thun, L Nghin cu sinh tng hp v tch chit cht
Th Thanh Xun, khng khun t chng nm si Penicillium
ng Th Mai Anh, waksmani Zaleski T10.6, phn lp t trm
Phm Cng Hot, tch bin Bnh nh. Hi ngh Khoa hc ton
quc ln th IV - Ha sinh v sinh hc phn
Chu Vn Minh
t phc v nng, sinh, y hc v cng nghip
thc phm, 2008, 892-896.

7. Danh mc cc lun n
TT Tn hc vin
Tn lun n
1 Nguyn Th Kim Nghin cu thnh phn ha hc v hot tnh
Thy
gy c t bo ca loi hi min Gellius
varius. Lun vn thc s Ha hc, 2007.
2 Trn Anh Tun
Nghin cu thnh phn ha hc v hot tnh
sinh hc ca loi san h mm Sarcophyton
mililatensis, Lun vn thc s Ha hc,
2008.
3 Nguyn Vn Thanh
Nghin cu thnh phn ho hc v kho st
hot tnh gy c t bo in vitro ca loi hi
sm Holothuria scabra v sao bin
Archaster typicus sinh sng ti vng bin
vit nam, Lun n Tin s Ho hc, 2008
4 Trn Thu H
Nghin cu thnh phn ha hc v hot tnh
sinh hc mt s loi hi min sinh sng ti
vng bin Vit Nam, Lun n Tin s Ha
hc, 2008-2012.
Ngoi ra ti t chc 01 hi tho:
-Hi tho khoa hc ti KC09.09/06-10 thng 12 nm 2008 ti H Ni
159

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ng

Ghi ch
bo
v
bo
v

bo
v

ang
thc
hin