Nui cy sm ngc linh

thu nhn gisenosides

Thnh vin:
ng Xun Phong
Trn Vn Phong
Nguyn Hong n
Nguyn Duy i

 I. Tng quan v sm ngc linh

II. Thnh phn ha hc sm ngc linh
III. Quy trnh nui cy sm ngc linh
IV. Thu nhn ginsenosides

 I. Tng quan v sm ngc linh

1.Danh php
Tn khoa hc : Panax vietnamesis Ha et Grushy

Gii (regnum): Plantae

Ngnh (divisio): Magnoliophyta
Lp (class):Magnoliopsida
B (ordo): Apiales
H (familia): Araliaceae
Phn h (subfamilia): Aralioideae
Chi (genus): Panax
Phn chi (subgenus): Panax
Nhnh (section): Panax
Loi (species): P. vietnamensis

2. c im sm ngc linh
- Sm Ngc Linh mc ti vng ni Ngc Linh thuc hai tnh Kon
Tum v Qung Nam trn cao 1.500 - 2.200 m.

- Sm mc tp trung di chn ni Ngc Linh, mt ngn ni cao

2.578m vi lp t vng trn granit dy trn 50cm, c

mn cao, ti xp v rng nguyn sinh cn rng,nhit thch
hp vo ban ngy l 20C-25C, ban m l :15C -18C
y l mt loi cy thn tho sng lu nm, cao 40cm n
100cm
Sm Ngc Linh c dng thn kh sinh thng ng, mu lc
hoc hi tm, nh, c ng knh thn 4-8mm, thng tn
li hng nm tuy thnh thong cng tn ti mt vi thn trong
vi nm. Thn r c ng knh 1-2cm, mc b ngang nh c
hong tinh trn hoc di mt t 1-3cm, mang nhiu r
nhnh v c.

II. Thnh phn ha hc ca sm.

1. Thnh phn ha hc ca sm
- Sm Ngc Linh cha ch yu cc saponin
hay gi l ginsenosides (saponin c cu
trc Damaran(vi hai i din l nhm
protopanaxadiol v nhm protopanaxadiol)
v saponin c cu trc olean)
- Ngoi ra cn 14 axit bo,16 axit amin
(trong c 8 acid amin khng thay th),
18 nguyn t a lng v vi lng.
- Sm Vit Nam mi c hp cht saponin
dammaran kiu ocotillol vi Majonosid R2
chim hn 50% hm lng saponin

Cng thc ca Majonosid R2

2. Tc ng ca sm n sc khe con ngi.

- Tc dng b chung, tng lc v sinh thch
nghi (antistress), phc hi s suy gim
chc nng gip cho tnh trng ca c th
tr li bnh thng m khi nim ca y hc
c truyn gi l "hi dng"
- Tc dng chng lo ha, khng cc c t
gy hi t bo, gip ko di s sng ca t
bo, tng to cc t bo mi
- Tc dng kch thch iu ha c ch min
dch, tng sc khng ca c th, phng
v chng li mt s bnh ung th ( Rh2 v
Rg3 ).

III. Quy trnh nui cy sm ngc linh

Ct mu

Kh trng

Nui trong mi
trng MS

Callus
( M so u tin)

H thng nui cy sinh hc

(Bioreactor)

Nui trong Erlen 100ml

Thu hi sn phm

-Ta chn nhng r sm trng th nh

(khong trn 5 nm) ct r l m mu. Kh
trng mu vi ethanol 70% sau ngm
ton b mu trong dung dch sodium
hypochlorite khong 15 ph t, cui cng ra
di vi nc ct khong 5 ph t.
- cho mu vo trong a petri cha mi
trng MS c b sung thm 0.5mg/l 6benzyl-aminopurine (BAP), 3mg/l 1naphaleneacetic acid (NAA) v 30g ng
sucrose, nui trong iu kin nhit 25 0C
trong ti. c bit pH ca mi trng phi
iu chnh mc 5.8 trc khi em kh
trng, kh trng 1210C trong 15 pht.

- khi m so c hnh thnh. Ct mo so

chuyn sang mi trng SH. Kch thc m
so ct khong 1cm, cho vo erlen cha
khong 40ml mi trng SH. ng thi mi
trng SH b sung thm 0.5mg/l BAP,
3mg/l NAA, 30g/l ng sucrose. a erlen
vo nui trong ti nhit l 250C vi tc
lc l 60 vng/pht.
- pH ca mi trng ta c th iu chnh
5.0, 5.5, 6.5, 7.0, trc khi a mi trng
vo kh trng. Nng ng, nitrogen,
photphate iu chnh khong 1-9%, 127.61148 mg/l v 26.9-242.1mg/l.

M so hnh thnh:

- - Ngoi ra ta c th

nui m so trong
bioreacor. Ct m
so mt di khong
2mm ng knh
0.5cm cho vo
bioreactor cha
mi trng SH.Tin
hnh nui trong iu
kin nhit l
250C, m 7080%, cng chiu
sng l 45 mol.m-2.s1.vi chu k chiu
sng l 10h/ ngy.

 Thnh phn mi trng MS(1962)

Thnh phn khong a lng:

NH4NO31650mg./l
KNO31900mg/l
CaCl2.2H20440mg./l
MgSO4.7H2O..370mg./l
KH2SO4.170mg/l
Thnh phn khong vi lng:
MnSO4.H2O.23,3mg/l
ZnSO4.7H2O8,6mg/l
H3BO3.6,2mg/l
KI.0,83mg/l
Na2MoO4.2H2O.0,25mg/l
CuSO4.5H2O0,025mg/./
CoCl2.6H2O.0,025mg/l
Na2EDTA.37,30mg/l

FeSO4.7H2O27,8mg/l
Vitamin v amino acid
Thiamine HCL ..0,1mg/l
Acid nicotinic.0,5mg/l
Pyridoxine HCL.0,5mg/l
Glycine..2,00mg/l

Thnh phn mi trng SH

2-Mi trng SH(1972):
Thnh phn khong a lng:
KNO32500mg/l
CaCl2.2H20200mg./l
MgSO4.7H2O..400mg./l
NH4H2PO4.300mg/l
Thnh phn khong vi lng:
MnSO4.H2O.10mg/l
ZnSO4.7H2O1,00mg/l
H3BO3.5,00mg/l
KI.1,00mg/l
Na2MoO4.2H2O.0,1mg/l
CuSO4.5H2O0,20mg/./
CoCl2.6H2O.0,1mg/l
Na2EDTA.20,00mg/l
FeSO4.7H2O15,00mg/l

Vitamin v amino acid:

Thiamine HCL ..5,00mg/l
Acid nicotinic.0,5mg/l
Pyridoxine HCL.0,5mg/l

Cc yu t nh hng n hm lng
saponin
a. pH

b. Nng ng

c. Nng nitrogen

d. Nng photphate

IV. Thu nhn ginsenosides

2kg mu
dung dch methanol 80%
lm bay hi
huyn ph bng nc
cho ete vo(ethyl ete)
lm bay hi lp
nc
chuyn vo ct sc k
Cc ging:
1.Nc
2.30% KOH (7g)
3.50% KOH (13g)
4.80% KOH(174g)
5.KOH + aceton

-Qu trnh tch chit KOH 80% th thu

c hm lng cao nht saponin.
+ 20(s) protopanaxdiol
+ 20(s) protopanaxtriol
+ ocotillol