coeus.vn a14.PHẨM-MÀU

CNG HA X HI CH NGHA VIT NAM
QCVN 4-10 : 2010/BYT
QUY CHUN K THUT QUC GIA

V PH GIA THC PHM - PHM MU
National technical regulation
on Food Additives - Colours
H NI - 2010
Li ni u
QCVN 4-10 :2010/BYT do Ban son tho quy chun k
thut quc gia v Ph gia thc phm v cht h tr ch
bin bin son, Cc An ton v sinh thc phm trnh duyt
v c ban hnh theo Thng t s 27/2010/TT-BYT
ngy 20 thng 5 nm 2010 ca B trng B Y t.
QUY CHUN K THUT QUC GIA

V PH GIA THC PHM - PHM MU
National technical regulation on Food Additives - Colours
I. QUY NH CHUNG
1. Phm vi iu chnh
Quy chun k thut quc gia (sau y gi tt l Quy chun) ny quy nh cc yu cu
k thut v qun l v cht lng, v sinh an ton i vi cc phm mu c s dng vi
mc ch lm ph gia thc phm.
2. i tng p dng
Quy chun ny p dng i vi:
2.1. T chc, c nhn nhp khu, xut khu, sn xut, bun bn v s dng cc
phm mu lm ph gia thc phm (sau y gi tt l t chc, c nhn).
2.2. C quan qun l nh nc c lin quan.
3. Gii thch t ng v ch vit tt:
3.1. Phm mu: l ph gia thc phm c s dng vi mc ch tng cng
hoc khi phc mu cho sn phm thc phm.
3.2. JECFA monograph 1 - Vol. 4 (JECFA monographs 1 - Combined
compendium of food addiditive specifications; Joint FAO/WHO expert committee on
food additives; Volume 4 - Analytical methods, test procedures and laboratory
solutions used by and referenced in the food additive specifications; FAO, 2006): Cc
yu cu k thut i vi ph gia thc phm, Tp 4 Cc phng php phn tch, quy
trnh th nghim, dung dch th nghim c s dng (hoc tham chiu) trong yu
cu k thut i vi ph gia thc phm; JECFA bin son; FAO ban hnh nm 2006.
3.3. M s C.A.S (Chemical Abstracts Service): M s ng k ha cht ca
Hip hi Ha cht Hoa K.
3.4. TS (test solution): Dung dch thuc th.
3.5. ADI (Acceptable daily intake): Lng n vo hng ngy chp nhn c.
3.6. INS (International numbering system): H thng m s quc t v ph gia
thc phm.
II. YU CU K THUT V PHNG PHP TH V LY MU
1. Yu cu k thut v phng php th i vi phm mu c quy nh ti
cc ph lc ban hnh km theo Quy chun ny nh sau :
1.1.
Ph lc 1:
Quy nh k thut v phng php th i vi

curcumin
1.2.
Ph lc 2:
1.3.
Ph lc 3:

riboflavin
Quy nh k thut v phng php th i vi tatrazin
1.4.
Ph lc 4:
Quy nh k thut v phng php th i vi quinolin
1.5.
Ph lc 5:
1.6.
Ph lc 6:
Quy nh k thut v phng php th i vi sunset

yellow FCF
Quy nh k thut v phng php th i vi carmin
1.7.
Ph lc 7:
1.8.
Ph lc 8:
1.9.
Ph lc 9:
1.10.
Ph lc 10:
1.11.
Ph lc 11:
1.12.
Ph lc 12:
1.13.
Ph lc 13:
1.14.
Ph lc 14:
1.15.
Ph lc 15:
1.16.
Ph lc 16:
1.17.
Ph lc 17:
1.18.
Ph lc 18:
1.19.
Ph lc 19:
1.20.
Ph lc 20:
1.21.
Ph lc 21:
1.22.
Ph lc 22:
1.23.
Ph lc 23:
1.24.
Ph lc 24:
1.25.
Ph lc 25:
1.26.
Ph lc 26:

carmoisin
amaranth
ponceau 4R
erythrosin
Quy nh k thut v phng php th i vi red 2G
Quy nh k thut v phng php th i vi allura
red AC
Quy nh k thut v phng php th i vi indigotin
Quy nh k thut v phng php th i vi brilliant
blue FCF
clorophyl
clorophyl phc ng
mui Natri, Kali ca Clorophyl phc ng
Quy nh k thut v phng php th i vi green S
caramen
Quy nh k thut v phng php th i vi brilliant
black PN
Quy nh k thut v phng php th i vi brown
HT
Quy nh k thut v phng php th i vi caroten tng hp
Quy nh k thut v phng php th i vi caroten
thc vt
Quy nh k thut v phng php th i vi cao
annatto
Quy nh k thut v phng php th i vi -Apocarotenal
Quy nh k thut v phng php th i vi este
methyl (hoc Ethyl) ca acid -Apo-8'-Carotenoic
1.27.
Ph lc 27:
1.28.
Ph lc 28:
1.29.
Ph lc 29:
1.30.
Ph lc 30:
Quy nh k thut v phng php th i vi cao v

nho
Quy nh k thut v phng php th i vi dioxyd
titan
Quy nh k thut v phng php th i vi oxyd st
Quy nh k thut v phng php th i vi fast
green FCF
2. Cc yu cu k thut quy nh trong Quy chun ny c th theo JECFA

monograph 1 - Vol. 4, ngoi tr mt s php th ring c m t trong cc ph lc.
Cc phng php th c hng dn trong Quy chun ny khng bt buc phi
p dng, c th s dng cc phng php th khc tng ng.
3. Ly mu theo hng dn ti Thng t 16/2009/TT-BKHCN ngy 02 thng 6
nm 2009 ca B Khoa hc v Cng ngh v hng dn kim tra nh nc v cht
lng hng ha lu thng trn th trng v cc quy nh khc ca php lut c lin
quan.
III. YU CU QUN L
1. Cng b hp quy
1.1. Cc phm mu phi c cng b ph hp vi cc quy nh ti Quy
chun ny.
1.2. Phng thc, trnh t, th tc cng b hp quy c thc hin theo Quy
nh v chng nhn hp chun, chng nhn hp quy v cng b hp chun, cng
b hp quy ban hnh km theo Quyt nh s 24/2007/Q-BKHCN ngy 28 thng 9
nm 2007 ca B trng B Khoa hc v Cng ngh v cc quy nh ca php lut.
2. Kim tra i vi phm mu
Vic kim tra cht lng, v sinh an ton i vi cc phm mu phi thc hin
theo cc quy nh ca php lut.
IV. TRCH NHIM CA T CHC, C NHN
1. T chc, c nhn phi cng b hp quy ph hp vi cc quy nh k
thut ti Quy chun ny, ng k bn cng b hp quy ti Cc An ton v sinh thc
phm v bo m cht lng, v sinh an ton theo ng ni dung cng b.
2. T chc, c nhn ch c nhp khu, xut khu, sn xut, bun bn v s dng
cc phm mu sau khi hon tt ng k bn cng b hp quy v bo m cht lng, v
sinh an ton, ghi nhn ph hp vi cc quy nh ca php lut.
V. T CHC THC HIN

1. Giao Cc An ton v sinh thc phm ch tr, phi hp vi cc c quan chc
nng c lin quan hng dn trin khai v t chc vic thc hin Quy chun ny.
2. Cn c vo yu cu qun l, Cc An ton v sinh thc phm c trch nhim
kin ngh B Y t sa i, b sung Quy chun ny.
3. Trng hp hng dn ca quc t v phng php th v cc quy nh
ca php lut vin dn trong Quy chun ny c sa i, b sung hoc thay th th
p dng theo vn bn mi.
Ph lc 1
YU CU K THUT V PHNG PHP TH I VI CURCUMIN
1. Tn khc, ch s
Vng ngh, Kurkum

INS: 100i
ADI = 0 3 mg/kg th trng.
2. nh ngha
Curcumin c sn xut bng cch chit c ngh - thn

r ca cy Curcuma longa L. (Curcuma domestica
Valeton) bng dung mi. thu c bt curcumin c
hm lng cao, dch chit c tinh ch bng phng
php kt tinh. Sn phm thu c cha cc curcumin,
thnh phn mu chnh l 1,7-bis-(4-hydroxy-3-methoxyphenyl)-hepta-1,6-dien-3,5-dion (tn khc: Curcumin,
Diferuloylmethan, CI Natural Yellow 3, CI (1975) 75300)
v dn cht desmethoxy- v bis-desmethoxy- ca n
theo t l khng c nh. Trong ch phm c th cha
lng nh du v nha t nguyn liu. Ch cc dung
mi sau y c th c s dng trong qu trnh chit
v tinh ch curcumin: Aceton, methanol, ethanol, isopropanol, hexan v ethyl acetat. Carbon dioxyd siu ti
hn cng c th c s dng trong qu trnh chit.
Tn ha hc
Cc thnh phn mu chnh:
M s C.A.S.
Cng thc ha hc
I.
1,7-Bis-(4-hydroxy-3-methoxyphenyl)-hepta-1,6dien-3,5-dion
II.
1-(4-Hydroxyphenyl)-7-(4-hydroxy-3methoxyphenyl)-hepta-1,6-dien-3,5-dion
III.
1,7-Bis-(4-hydroxyphenyl)-hepta-1,6-dien-3,5-dion
I.
458-37-7
II.
33171-16-3
III.
33171-05-0
I.
C21H20O6
II.
C20H18O5
III.
C19H16O4
I.
R1= R2=OCH3
II.
R1= OCH3; R2=H
III.
R1= R2=H
Cng thc cu to
Khi lng phn t
I.
368,39
II.
338,39
III.
308,39
3. Cm quan
Bt tinh th mu vng cam.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc v diethyl ether; tan trong ethanol

v acid acetic bng.
Cc phn ng mu
Dung dch mu th trong ethanol c mu vng v cho

hunh quang xanh nht. Thm dung dch mu th ny
vo acid sulfuric c s c mu thm.
Sc k bn mng
t yu cu (m t trong phn Phng php th).
5.2. tinh khit

Tn d dung mi
Aceton:
Khng c qu 30,0 mg/kg
Hexan:
Methanol:
Ethanol:
Isopropanol:
Ethyl acetat:
Ch
Khng c qu 2,0 mg/kg
5.3. Hm lng
Khng nh hn 90,0% tng cc thnh phn mu.
6. Phng php th
6.1. nh tnh
Cc phn ng mu
X l dung dch mu th trong nc hoc trong ethanol

long vi acid hydrocloric n khi mu cam bt u xut
hin. Chia dung dch sau khi x l thnh 2 phn, thm
bt (hoc tinh th) acid boric vo 1 phn. Mu hng
s xut hin nhanh sau khi thm, mu s d nhn bit
hn khi so vi phn khng thm acid boric. Phn ng c
th tin hnh bng cch nhng mt mu giy lc vo
dch mu th tan trong ethanol, lm kh ti 100 o, sau
tm t mu giy lc bng dung dch acid boric long
thm vi git acid hydrocloric. kh, s xut hin mu
hng.
Sc k bn mng
Chm 5 L dch th (0,01 g mu th trong 1 ml ethanol

95%) trn 1 bn mng sc k (cellulose vi tinh th; 0,1
mm). Khai trin sc k trong bnh cha hn hp dung
mi pha ng 3-methyl-1-butanol/ethanol/nc/dung
dch amoniac (4:4:2:1), dung mi khai trin trn bn

mng 10-15 cm t vch xut pht. Kim tra, quan st
di nh sng thng v n UV:
Di nh sng thng v UV c 2 hoc 3 vt mu vng
vi Rf trong khong 0,2 - 0,4.
Di n UV c cc vt vi Rf khong 0,6 v 0,8
Cc vt u c hunh quang mu vng di n UV.
6.2 tinh khit
Ch
Xc nh bng k thut hp th nguyn t thch hp cho

hm lng quy nh. La chn c mu th v phng
php chun b mu da trn nguyn tc ca phng
php m t trong JECFA monograph 1-Vol. 4 phn cc
phng php phn tch cng c.
6.3. nh lng
Cn 0,08 g mu th (chnh xc n mg), chuyn vo
bnh nh mc 200 ml v ho tan bng lc vi ethanol,
nh mc n vch bng ethanol. Dng pipet ht 1 ml
dung dch v cho vo bnh nh mc 100 ml, nh mc
n vch bng ethanol.
Xc nh hp th quang ti 425 nm, cuvet o dy 1
cm. Tnh t l tng hm lng cht mu trong mu th
theo cng thc:
Trong :
A = hp th ca dung dch mu th
W = Khi lng mu th
1607 = hp th ring ca dung dch curcumin chun
trong ethanol ti 425 nm.
Vic xc nh hp th quang phi tin hnh ngay khi
pha xong dung dch, bi v mu ca dung dch c th
nht dn theo thi gian.
Ph lc 2
YU CU K THUT V PHNG PHP TH I VI RIBOFLAVIN
1. Tn khc, ch s
Vitamin B2, Lactoflavin

INS: 101i
ADI = 0 - 0,5 mg/kg th trng.
2. nh ngha
Tn ha hc
Riboflavin; 3,10-dihydro-7,8-dimethyl-10-[(2S,3S,4R)2,3,4,5-tetrahydroxypentyl]benzo-[g]pteridin-2,4-dion; 7,8dimethyl-10-(1'-Dribityl) isoalloxazin.
M s C.A.S.
83-88-5
Cng thc ha hc
C17H20N4O6
Cng thc cu to
CH2OH
HO C
HO C
HO C
CH2
H3C
H3C
O
NH
Khi lng phn t
376,37
3. Cm quan
Bt tinh th mu vng n vng cam, c mi nh.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Rt kh tan trong nc, thc t khng tan trong cn, ether,

aceton v cloroform; rt d tan trong dung dch kim long.
Quang ph
S dng dung dch mu th trong nc, chun b trong

phn nh lng, xc nh hp th quang (A) ti cc
bc sng 267 nm, 375 nm, 444 nm.
T s A375/A267 nm trong khong 0,31 v 0,33.
Gc quay cc ring
[ ]20,D: trong khong -115o v -140o.
Phn ng mu
5.2. tinh khit

Gim khi lng khi lm
kh
Khng c qu 1,5%.
Tro sulfat
Khng c qu 0,1%.
Cc cht mu ph
Cc amin thm bc nht
Khng c qu 100,0 mg/kg tnh theo anilin.
Ch
5.3. Hm lng
C17H20N4O6
6. Phng php th
Khng c qu 2,0 mg/kg.

Khng c thp hn 98,0%.
6.1 nh tnh
Gc quay cc ring
Phn ng mu
Sy kh mu ti 100 oC trong 4 g. Cn 50 mg v ha tan

trong dung dch natri hydroxyd 0,05 N khng c carbonat v
pha long n 10 ml cng bng dung dch ny. o
quay quang ca dung dch trong 30 pht k t khi pha dung
dch.
Ha tan khong 1 mg mu vo 100 ml nc. Dung dch c
mu vng xanh nht khi cho nh sng truyn qua v c
hunh quang xanh vng m vi nh sng phn x. Mu
s b mt khi thm acid v c hoc kim v c.
6.2. tinh khit

Gim khi lng khi lm
kh
Sy ti 105 oC, trong 4 gi
Tro sulfat
- Th theo JECFA monograph 1 - vol.4

- Mu th: cn 2 g
Cc cht mu ph
Chun b dung dch chun khng c lumiflavin cho th

nghim bng cch pha long 3 ml dung dch kali dicromat
0,1 N vi nc n 1000 ml. Cho mt t cloroform qua
ct nhm oxyd loi ht ethanol. Ly 10 ml cloroform ny,
thm 35 mg mu, lc trong 5 pht v lc. Mt mu ca
dch lc khng c m hn mt mu ca 10 ml chun
khi quan st trong dng c cha cng loi
Ch
Th theo hng dn ti JECFA monograph 1 - vol.4. S

dng k thut hp th nguyn t thch hp vi hm lng
ch nh xc nh. La chn c mu th v phng
php chun b mu da trn nguyn tc ca phng php
m t trong Ph lc 1: Cc phng php th chung phn
cc phng php phn tch cng c
6.3. nh lng
Tin hnh nh lng trong mi trng t nh sng. Phn
tn 65,0 mg mu th trong 5 ml nc trong bnh nh mc
500 ml mu nu, cn m bo rng ton b mu th b
tm t, ha tan trong 5 ml dung dch natri hydroxyd 2 N.
Ngay khi mu tan hon ton, thm 100 ml nc v 2,5 ml
acid acetic bng v pha long bng nc n 500 ml.
Ly 20,0 ml dung dch ny, thm 3,5 ml dung dch natri
acetat 1,4% (kl/tt) v pha long vi nc n 200 ml.
o hp th quang (A) ti cc i hp th 444 nm.
% Riboflavin = [(A5000) / (328W)] 1,367
trong
A = hp th ca dung dch mu th ti 444 nm
W = Khi lng mu th, tnh bng g.
Ph lc 3
YU CU K THUT V PHNG PHP TH I VI TARTRAZIN
1. Tn khc, ch s
CI Food Yellow 4, FD&C Yellow No. 5; S CI (1975): 19140

INS: 102
2. nh ngha
Ch yu gm Trinatri 5-hydroxy-1-(4-sulfonphenyl)-4-(4sulfonphenylazo)-H-pyrazol-3-carboxylat cng cc cht

mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh
phn khng mu chnh.
C th chuyn thnh cht mu nhm (aluminium lake)
tng ng, trong trng hp ny p dng quy nh chung
i vi cc loi cht mu nhm.
Tn ha hc
Trinatri 5-hydroxy-1-(4-sulfonphenyl)-4-(4-sulfonphenylazo)H-pyrazol-3-carboxylat
M s C.A.S.
1934-21-0
Cng thc ha hc
C16H9N4Na3O9S2
Cng thc cu to
NaOOC
N
N
NaO3S
N N
SO3Na
OH
Khi lng phn t
534,37
3. Cm quan
Dng bt hoc ht mu cam nht.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, t tan trong ethanol.
nh tnh cc cht mu
Phi c phn ng c trng ca cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC
Khng c qu 15,0% cng vi clorid v sulfat tnh theo

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 1,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% (tng cc acid Tetrahydroxysuccinic;

acid 4-Hydrazinbenzensulfonic;
acid 4-Aminobenzensulfonic; acid 5-Oxo-1-(4-sulfophenyl)2-pyrazolin-3-carboxylic; acid 4,4'-Diazoaminodi (acid
benzensulfonic).
Cc amin thm bc nht Khng c qu 0,01% tnh theo anilin.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
Khng c thp hn 85,0% tng cc cht mu.
6. Phng php th
6.1. tinh khit
Ch
- Th theo hng dn ti JECFA monograph 1-Vol. 4.

- S dng k thut hp th nguyn t thch hp vi hm
lng ch nh xc nh. La chn c mu th v
phng php chun b mu da trn nguyn tc ca
phng php m t trong JECFA monograph 1-Vol. 4, Cc
phng php th chung phn cc phng php phn tch
cng c.
Cc cht mu ph
- Th theo hng dn ti JECFA monograph 1-Vol. 4

- S dng iu kin sau: Dung mi khai trin l dung mi
s 4; chiu cao ca tuyn dung mi ~ 12 cm.
Cc cht hu c ngoi
cht mu

- S dng k thut HPLC vi cc iu kin nh sau: HPLC
ra gii gradient 2 n 100% vi tc tng c nh
2%/pht.
6.2. nh lng
Tin hnh theo hng dn trong chuyn lun xc nh tng
hm lng bng chun vi Titan (III) clorid (trong Ph
lc 1: Cc phng php th chung) nh sau:
Cn 0,6 - 0,7 g mu th;
m 15 g natri hydro tartrat;
Mi ml TiCl3 0,1 N tng ng vi 13,56 mg cc cht
mu (D).
Ph lc 4
YU CU K THUT V PHNG PHP TH I VI QUINOLIN
1. Tn khc, ch s
CI food yellow 13, CI (1975) No. 47005,

INS: 104
2. nh ngha
Ch phm thu c bng cch sulfonat ho 2-(2-quinolyl)1,3-indandion hoc hn hp cha khong hai phn ba 2-(2quinolyl)-1,3-indandion v mt phn ba 2-[2-(6-methylquinolyl)]1,3-indandion; Ch phm cha hn hp mui natri
ca cc disulfonat (ch yu), cc monosulfonat and cc
trisulfonat ca cc hp cht nu trn v cc cht mu ph
cng vi natri clorid v/hoc natri sulfat l cc thnh phn
chnh khng to mu.
Quinolin c th chuyn thnh dng mu nhm tng ng,
trong trng hp ny p dng quy chun chung cho dng
mu nhm ca cc phm mu.
Tn ha hc
Dinatri 2-(1,3-dioxo-2-indanyl)-6,8-quinolin sulfat; dinatri 2(2-quinolyl)-indan-1,3-dion disulfonat (thnh phn chnh)
M s C.A.S.
8004-72-0 (acid disulfonic khng methyl ha)
Cng thc ha hc
C18H9NNa2O8S2 (thnh phn mu chnh)
Cng thc cu to
Thnh phn mu chnh

R1
O
N
R2
O
mui 6 : R1 = SO3Na, R2 = H
mui 8: R1, R2 = SO3H
Khi lng phn t
Thnh phn mu chnh: 477,38
3. Cm quan
Bt hoc ht nh mu vng.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, t tan trong ethanol
nh tnh cc cht mu
Phi c phn ng c trng ca cc cht mu.
5.2. tinh khit

Gim khi lng khi sy Khng c qu 30,0% cng vi clorid v sulfat tnh theo
mui natri.
ti 135o
Cht khng tan trong
nc
Khng c qu 0,2%.
Ch
Km
Cc cht mu ph
Tng hm lng 2 cht 2-(2-quinolyl)-1,3-indandion v 2[2-(6-methylquinolyl)]-1,3-indandion khng c qu 4,0

mg/kg
Cc hp cht hu c
ngoi cc thnh phn
mu
Tng hm lng cc cht 2-methylquinolin, acid 2methylquinolinsulfonic, acid phtalic, 2,6-dimethylquinolin,

acid 2,6-dimethylquinolin sulfonic khng c qu 0,5%
Cc amin thn bc nht Khng c qu 0,01% tnh theo anilin.

khng sulfonat ha
Cht c th chit bng
ether
Khng c qu 0,2%.
5.3. Hm lng
Khng nh hn 70% tng cc thnh phn mu. i vi ch

phm quinolin vng sn xut t 2-(2-quinolyl)-1,3-indandion
s c cc thnh phn sau:
- Khng nh hn 80,0% dinatri 2-(2-quinolyl)-indan- 1,3diondisulfonat;
- Khng c qu 15,0% natri 2-(2-quinolyl)-indan-1,3dionmonosulfonat;
- Khng c qu 7,0% trinatri 2-(2-quinolyl)-indan-1,3diontrisulfonat;
6. Phng php th
6.1. tinh khit
Ch
Th theo hng dn ti JECFA monograph 1-Vol. 4. S

m t trong JECFA monograph 1-Vol. 4, Cc phng php
th chung phn cc phng php phn tch cng c.
Cc cht mu ph
Th gii hn ca 2-(2-quinolyl)-1,3-indandion v 2-[2-(6methylquinolyl)]-1,3-indandion.

S dng thit b v ether c cht lng theo m t trong
chuyn lun xc nh cht c th chit bng ether v tin
hnh chit nh sau: Ra dch chit 2 ln mi ln vi 25 ml
nc ct. Cho ether bay hi dch chit cn khong 5 ml,
sau cho vo t sy ti nhit 105o ui ht ether.
Ho tan cn cn li trong cloroform v pha long n chnh

xc 10 ml. Xc dnh hp th quang ca dung dch ny ti
bc sng hp th cc i (khong 420 nm) s dng
cloroform lm dung dch i chng. Gi tr hp th
quang = 0,27 tng ng vi gii hn ca 2-(2-quinolyl)-1,3indandion l 4 mg/kg. 2-[2-(6-methylquinolyl)]-1,3-indandion
c nh gi nh 2-(2-quinolyl)-1,3-indandion.
Cc hp cht hu c
ngoi cc thnh phn
mu
Cc hp cht hu c ngoi cc thnh phn mu trong

quinolin ch c trong ch phm sn xut t 2-(2-quinolyl)1,3-indandion.
Xc nh bng phng php sc k lng, vi cc iu kin
nh sau:
Thit b: Sc k lng phn gii cao c b iu khin gradien
nng pha ng.
Detector: Detector UV cho HPLC c th o hp th
quang ti 254 nm.
Ct: Nucleosil, 250X4 mm, 7 m.
H dung mi:
A: m acetat pH 4,6:nc (1:10). m acetat bao gm
dung dch NaOH 1M : acid acetic 1 M : nc (5:10:35).
B: (A):methanol (20:80).
Nng mu 1%(m/v) trong dung mi A.
iu kin gradient nng pha ng:
Pht
%A
%B
100
15
65
35
20
50
50
25
100
36
100
42
100
Tc dng: 1 ml/pht
6.2. nh lng
Tin hnh theo hng dn trong chuyn lun Xc nh tng
hm lng bng phng php quang ph (JECFA
monograph 1 - Vol. 4).
Dung mi: m photphat pH 7
Pha long dung dch A: t 10 ml pha n 250 ml
hp th ring (a): 86,35

Bc sng hp th cc i: 415 nm
Ring i vi ch phm quinolin sn xut t 2-(2-quinolyl)1,3-indandion xc nh t l % ca di-, mono-, trisulfonat:
S dng iu kin HPLC m t trong chuyn lun xc nh
cc hp cht hu c ngoi thnh phn mu, vi nng
dung dch mu th trong dung mi HPLC A l 0,05% thay
cho 1%. Biu th kt qu theo t l % tng cc thnh phn
mu c trong ch phm.
Ph lc 5
YU CU K THUT V PHNG PHP TH I VI SUNSET YELLOW FCF
1. Tn khc, ch s
CI Food Yellow 3, FD&C Yellow No. 6; S CI (1975): 15985

INS: 110
2. nh ngha
Ch yu gm dinatri 6-hydroxy-5-(4-sulfonphenylazo)-2naphthalen-6-sulfonat cng cc cht mu ph cng vi

NaCl v (hoc) Na2SO4 l cc thnh phn khng mu
chnh.
Tn ha hc
dinatri 6-hydroxy-5-(4-sulfonphenylazo)-2-naphthalen-6sulfonat.
M s C.A.S.
2783-94-0
Cng thc ha hc
C16H10N2Na2O7S2
Cng thc cu to
HO
NaO3S
N N
SO3Na
Khi lng phn t
452,38
3. Cm quan
Dng bt hoc ht mu cam.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
Phi c phn ng c trng ca cc cht mu.
5.2. tinh khit

Gim khi lng khi lm
kh ti 135oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 5,0%.
Trong khng c qu 2,0% cc cht mu khc ngoi

trinatri 2-hydroxy-1-(4-sulfonphenylazo)naphthalen-3,6disulfonat.
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% tng cc acid 4-amino-1benzensulfonic;

Acid 3-hydroxy-2,7-naphthalendisulfonic;
Acid 6-hydroxy-2-naphthalensulfonic;
Acid 7-hydroxy-1,3-naphthalendisulfonic;
Acid 4,4'-diazoaminodibenzensulfonic;
Acid 6,6'-dioxy-2-naphthalensulfonic.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Ch

phng php m t trong JECFA monograph 1-Vol. 4 phn
cc phng php phn tch cng c.
Cc cht mu ph

Cc cht hu c ngoi
cht mu

4%/pht, tip theo ra gii ti 100%.
6.2. nh lng
hm lng bng chun vi Titan (III) clorid (trong
JECFA monograph 1-Vol. 4) nh sau:
Cn 0,5 - 0,6 g mu th;
m 10 g natri citrat;
mu (D).
QUY
Ph lc 6
YU CU K THUT V PHNG PHP TH I VI CARMIN
1. Tn khc, ch s
Cochineal carmin, CI Natural Red 4, CI (1975) No.

75470,
INS 120
ADI cho c nhm carmin l 0 - 5 mg/kg th trng tnh
theo amoni carminat, cc mui Ca, K, Na quy ra tng
ng.
2. nh ngha
Ch phm thu c bng cch chit t phm yn chi

cha xc kh ca rp son ci (Dactylopius coccus
Costa) vi nc; Cht mu chnh l mt phc chelat
nhm ngm nc ca acid carminic, trong phc ny t l
phn t gia nhm v acid carminic l 1/2.
Trong ch phm thng mi, cht mu chnh kt hp
vi cation amoni; calci; kali hoc natri, dng t do hay
kt hp, cc cation ny lun d. Trong ch phm c th
cha protein t xc cn trng nguyn liu, c th cha
mui carminat v lng d t cation nhm.
Tn ha hc
Phc chelat nhm ngm nc ca acid carminic (7-betaD-glucopyranosyl-3,5,6,8-tetrahydroxy-1-methyl-9,10dioxo-anthracen-2-carboxylic acid)
M s C.A.S.
1390-65-4 (carmin)
1260-17-9 (acid carminic)
Cng thc ha hc
Acid carminic C22H20O13
Cng thc cu to
Acid carminic
CH2OH
O H
H
OH
OH
H
OH
CH3
COOH
OH
HO
OH
OH
Phc nhm ca acid carminic
Khi lng phn t
Acid carminic: 492,39
3. Cm quan
Dng bt hoc mnh vn mu ti m
4. Chc nng
Phm mu
5. Yu cu k thut
5.1. nh tnh
tan
tan ca ch phm carmin ph thuc vo bn cht

ca cation c mt trong ch phm. Ch phm cha ch
yu cation amoni (mui amoni carminat) tan tt trong
nc ti pH 3,0 v pH 8,5. Ch phm cha ch yu
cation calci (mui calci carminat) t tan trong nc ti pH
3,0 nhng tan tt ti pH 8,5.
Phn ng mu
t yu cu (m t trong phn Phng php th)
5.2. tinh khit

Gim khi lng khi lm Khng c qu 20,0%.
kh
Tro ton phn
Khng c qu 12,0%.
Protein
Khng c qu 25,0%.
Cht khng tan trong Khng c qu 1,0%.

dung
dch
amoniac
long
Ch
Yu cu v vi sinh vt
Salmonella: m tnh.
5.3.
Hm
C22H20O13
lng Khng thp hn 50,0% C22H20O13 tnh theo ch phm

lm kh.
6. Phng php th
6.1 nh tnh
Phn ng mu
- Kim ha nh dung dch mu th bng cch thm 1

git dung dch natri hydroxyd (hoc kali hydroxyd) 10%.
Dung dch chuyn sang mu tm.
- Thm mt t tinh th natri dithionit (Na2S2O4) vo cc
dung dch mu th trung tnh, kim ha, acid ha.
Cc dung dch ny khng b mt mu.
- Lm kh lng nh mu th trn a s. ngui
hon ton, nh 1-2 git thuc th acid sulfuric vo cn
cn li trn a. Mu ca cn khng thay i.
- Acid ha dung dch mu th trong nc bng dung
dch thuc th acid hydrocloric, th tch thuc th bng
1/3 th tch dch th, cho thm ru amylic, lc u.
Tch ly lp ru amylic v ra 2-4 ln vi ng th tch
nc ct loi han ton acid hydrocloric. Pha long
phn ru amylic vi ether du ho (40-60 0) ng th
tch hoc th tch ether du ho gp i, lc vi ln vi
mt t nc ct loi ht mu. Thm, nh tng git
dung dch urani acetat 5%, lc u sau mi ln thm.
Dung dch chuyn sang mu xanh lc ngc c trng.
6.2 tinh khit

Gim khi lng khi lm - Th theo hng dn ti JECFA monograph 1-Vol. 4
- sy ti 135 oC trong 3 gi.
kh
Tro ton phn

- chuyn lun xc nh tro, cn 1 g mu.
Protein

- chuyn lun xc nh Nit phi amoni X 6,25
Cht khng tan trong Cn 0,25 g (chnh xc n mg) mu th c lm

dung
dch
amoniac kh, hon tan trong 2,5 ml dung dch amoniac long (160
ml dung dch thuc th amoniac c, thm nc ct n
long
500 ml) v pha long vi nc ct n 100 ml, thu c
dung dch trong. Lc dung dch ny qua phu thu tinh
xp. Ra phu vi dung dch amoniac 0,1% v lm kh
n khi lng khng i ti 105o.
Ch
Xc nh bng k thut hp th nguyn t thch hp cho

php chun b mu da trn nguyn tc ca phng
php m t trong JECFA monograph 1-Vol. 4 phn cc
phng php phn tch cng c.
6.3. nh lng
Cn 100 mg (chnh xc n mg) mu th, ho tan vo
30 ml dung dch acid hydrocloric 2N si, lm ngui.
Chuyn ton b dung dch sang bnh nh mc 1000 ml,
pha long v nh mc ti vch bng nc ct, lc u.
Xc nh hp th quang ca dung dch ti bc sng
hp th cc i khong 494 nm, s dng mu trng l
nc ct, cuvet o c dy 1 cm. Tnh hm lng %
acid carminic trong mu th theo cng thc:
(100 A 100) / (1,39 W)
trong :
A = hp th quang ca dung dch mu th;
W = Khi lng mu th,tnh bng mg;
1,39 = hp th quang ca dung dch acid carminic
nng 100 mg/1000 ml;
Nu hp th quang o c khng nm trong khong
gi tr 0,65 - 0,75, chun b mu th khc v hiu chnh
khi lng mu th thch hp.
Ph lc 7
YU CU K THUT V PHNG PHP TH I VI CARMOISIN
1. Tn khc, ch s
CI Food Red 3, Azorubine; CI (1975): 14720

INS: 122
ADI = 0 - 4 mg/kg th trng.
2. nh ngha
Ch yu gm dinatri 4-hydroxy-3-(4-sulfon-1-naphthylazo)1-naphthalensulfonat cng cc cht mu ph cng vi

chnh.
Tn ha hc
dinatri 4-hydroxy-3-(4-sulfon-1-naphthylazo)-1naphthalensulfonat.
M s C.A.S.
3567-69-9
Cng thc ha hc
C20H12N2Na2O7S2
Cng thc cu to
HO
NaSO3
N N
SO 3Na
Khi lng phn t
502,44
3. Cm quan
Dng bt hoc ht mu .
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
Phi c phn ng c trng ca cht mu.
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 1,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% tng cc acid 4-Amino-1naphthalensulfonic v acid 4-hydroxy-1-naphthalensulfonic.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1 tinh khit
Ch

Cc cht mu ph

Cc cht hu c ngoi
cht mu

ra gii gradient t 2% n 100% vi tc tng c nh
2%/pht.
6.3. nh lng
Cn 0,5 - 0,6 g mu th;
m 15 g natri hydrotartrat;
mu (D).
Ph lc 8
YU CU K THUT V PHNG PHP TH I VI AMARANTH
1. Tn khc, ch s
CI Food Red 9, Naphtol Rot S.; CI (1975): 16185

INS: 123
ADI = 0 - 0,5 mg/kg th trng
2. nh ngha
Ch yu gm trinatri 3-hydroxy-4-(4-sulfon-1-naphthylazo)2,7-naphthalendisulfonat v cc cht mu ph cng vi

chnh.
Tn ha hc
trinatri 3-hydroxy-4-(4-sulfon-1-naphthylazo)-2,7naphthalendisulfonat
M s C.A.S.
915-67-3
Cng thc ha hc
C20H11N2Na3O10S3
Cng thc cu to
HO
NaO3S
SO 3Na
N N
SO 3Na
Khi lng phn t
604,48
3. Cm quan
Dng bt hoc ht mu nu hoc nu sm.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 3,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% tng cc acid 4-amino-1naphthalensulfonic;

acid 3-hydroxy-2,7-naphthalendisulfonic; acid 6-hydroxy-2naphthalensulfonic; acid 7-hydroxy-1,3naphthalendisulfonic v acid 7-hydroxy-1,3,6- naphthalentrisulfonic.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1 tinh khit
Ch

Cc cht mu ph

s 3; chiu cao ca tuyn dung mi ~ 17 cm, sau 1 gi khai
trin.
Cc cht hu c ngoi
cht mu

4%/pht.
6.2. nh lng
Cn 0,7 - 0,8 g mu th;
mu (D).
Ph lc 9
YU CU K THUT V PHNG PHP TH I VI PONCEAU 4R
1. Tn khc, ch s
CI Food Red 7, Cochineal Red A ; New Cochineal;

CI (1975): 16255
INS: 124
2. nh ngha
Ch
yu
gm
trinatri
d-2-hydroxy-1-(4-sulfon-1naphthylazo)-6,8-naphthalendisulfonat cng cc cht mu
ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn
khng mu chnh.
Tn ha hc
Trinatri-2-hydroxy-1-(4-sulfon-1-naphthylazo)-6,8naphthalendisulfonat
M s C.A.S.
2611-82-7
Cng thc ha hc
C20H11N2Na3O10S3.1,5H2O
Cng thc cu to
HO
NaO3S
N N
NaO3S
SO3Na
Khi lng phn t
631,51
3. Cm quan
Dng bt hoc ht mu .
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 1,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% tng cc acid 4-amino-1naphthalensulfonic; acid 7-hydroxy-1,3naphthalendisulfonic; acid 3-hydroxy-2,7naphthalensulfonic; acid 6-hydroxy-2-naphthalensulfonic v
acid 7-hydroxy-1,3,6-naphthalentrisulfonic.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Ch

Cc cht mu ph

s 3; chiu cao ca tuyn dung mi ~ 17 cm, sau 1 gi khai
trin.
Cc cht hu c ngoi
cht mu

2%/pht.
6.2. nh lng
Cn 0,7 - 0,8 g mu th;
mu (D).
Ph lc 10
YU CU K THUT V PHNG PHP TH I VI ERYTHROSIN
1. Tn khc, ch s
CI Food Red 14, FD&C Red No. 3; CI (1975): 45430

INS 127
ADI = 0 - 0,1 mg/kg th trng
2. nh ngha
Ch yu l mui dinatri 9-(o-carboxyphenyl)-6-hydroxy2,4,5,7-tetraiodo-3-isoxanthon monohydrat cng cc cht

phn khng mu chnh.
Tn ha hc
Mui dinatri 9-(o-carboxyphenyl)-6-hydroxy-2,4,5,7tetraiodo-3-isoxanthon monohydrat
M s C.A.S.
16423-68-0
Cng thc ha hc
C20H6I4Na2O5.H2O
Cng thc cu to
I
O
I
O
2 Na
I
I
COO
Khi lng phn t
897,88
3. Cm quan
Dng bt hoc ht mu .
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc v trong ethanol
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Iod v c
Khng c qu 0,1% tnh theo natri iodid.
Cht khng tan trong

nc
Khng c qu 0,2%.
Km
Ch
Cc cht mu ph
Khng c qu 4,0% (khng bao gm Fluorescein).
Fluorescein
Cc cht hu c ngoi
cht mu
Khng c qu 0,2% Tri iodoresorcinol

Khng c qu 0,2% acid 2-(2,4-dihydroxy-3,5-diiodobenzoyl) benzoic
Cc cht c th chit
bng ether
Khng c qu 0,2% i vi dung dch mu th pH < 7.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Iod v c
Cn 1,0 g mu th vo mt cc c m 100 ml. Thm 75 ml

nc ct, dng my khuy t khuy cho tan hon ton.
Cho in cc c trng cho iod v mt in cc so snh
vo dung dch v t my o th thang o thch hp. c
th ca h (mV).
Dng buret thm dung dch bc nitrat 0,001 M, u tin mi
ln thm 0,5 ml, khi gn n im tng ng (nhn bit
bng s thay i in th sau mi ln thm) gim cn 0,1
ml/ln, n nh v c in th ca h sau mi ln thm
(mV). Tin hnh chun n khi thm dung dch bc
nitrat 0,001 M m khng lm thay i in th dung dch
nhiu. V ng biu th s ph thuc gia in th dung
dch (mV) vo th tch dung dch bc nitrat 0,001 M thm
vo. im tng ng c xc nh l im m th tch
dung dch bc nitrat 0,001 M thm vo tng ng vi
dc ca ng th ln nht.
Hm lng natri iodid trong mu c tnh = chun x
0,015%.
Ch

Cc cht mu ph

s 5; chiu cao ca tuyn dung mi ~ 17 cm. Ch : Khng
bn mng sc k trc tip ra ngoi snh sng mt tri
Cc cht hu c ngoi
cht mu

- S dng k thut sc k ct vi cc iu kin hp th nh
sau:
Tri-iodoresorcinol: 0,079 mgL-1cm-1 ti bc sng 233 nm,

trong mi trng acid.
Acid 2-(2,4-dihydroxy-3,5-di-iodobenzoyl) benzoic: 0,047
mgL-1cm-1 ti bc sng 348 nm, trong mi trng kim.
Fluorescein
Nguyn tc:
Fluorescein c tch khi mu th bng sc k lp mng
(TLC) v so snh vi sc k ca Fluorescein chun ti
nng tng ng vi gii hn quy nh.
Dung mi:
Methanol + nc + dung dch amoniac (khi lng ring ~
0,890) (500 ml + 400 ml + 100 ml).
Chun b mu:
Cn 1,0 g mu th, ha tan trong khong 50 ml dung mi
v pha long n 100 ml.
Chun:
Cn chnh xc lng Fluorescein ( tinh ch bng cch
kt tinh li trong ethanol) tng ng vi 1 g x hm lng
cc cht mu trong mu (xc nh trong phn nh lng).
Ha tan trong nc (hoc trong nc c 10 ml dung dch
amoniac c khi lng ring ~ 0,890 nu s dng acid
khng c Fluorescein), pha long v nh mc n 100
ml. Tin hnh pha long theo cc bc lin tip nh sau:
Ht 1ml pha long n 100 ml bng nc.
Ht 1ml pha long n 100 ml bng nc.
Ht 20 ml pha long n 100 ml bng dung mi.
Dung mi sc k:
n-Butanol + nc + dung dch amoniac (khi lng ring ~
0,890) + ethanol (100 ml + 44 ml + 1 ml + 22,5 ml).
Tin hnh:
Ln lt chm cnh nhau 25 L dung dch th v dung
dch chun trn cng 1 bn mng cellulose. Khai trin trong
h dung mi sc k trong 16 gi. kh bn mng. Soi
di n UV v so snh hunh quang ca vt th vi vt
chun. Mt hunh quang ca vt th khng c qu
vt chun.
Ch : khng bn mng ra nh sng mt tri trc tip.
6.2. nh lng
Cn 1 g (chnh xc n mg) mu th, ha tan trong 250 ml
nc, chuyn vo mt cc 500 ml sch, thm 8,0 ml acid
nitric 1,5 N, khuy u. Lc qua phu lc thy tinh xp (
xp 3, ng knh 5 cm), cha sn a thy tinh v cn
b. Ra cn bng acid nitric 0,5% n khi dch ra khng b
c khi th vi dung dch bc nitrat (TS), sau vi 30 ml
nc ct. Sy ti nhit 135 5 oC n khi lng khng
i. Cn thn ph cn bng a thy tinh, ngui trong
bnh ht m v cn.
Hm lng tng
cc cht mu =
Khi lng cn x 107,4

Khi lng mu th
Xc nh Cht khng tan trong acid hydrocloric trong mu

nhm erythrosin
Ha cht, thuc th:
Acid hydrocloric c
Acid hydrocloric 0,5% (tt/tt)
Dung dch amoniac long (pha long 10 ml dung dch
amoniac c khi lng ring =0,890 bng nc n 100
ml).
Tin hnh:
Cn khong 5 g (chnh xc n mg) mu th vo cc 500
ml. Thm 250 ml nc v 60 ml acid hydrocloric c. un
si ha tan nhm trong khi erythrosin chuyn v
dng acid t do, khng tan trong acid. Lc qua phu lc
thy tinh xp s 4 cn b. Ra cn vi mt lng nh
acid hydrocloric 0,5% nng, sau vi nc ct nng. Loi
b dch ra acid, ly phu thy tinh xp ra v ra phu
bng dung dch amoniac long n khi dch ra khng c
mu. Sy ti 135 oC n khi lng khng i. Cn phu
v tnh khi lng cn cn li v tnh hm lng % so vi
khi lng mu ly.
Ph lc 11
YU CU K THUT V PHNG PHP TH I VI RED 2G
1. Tn khc, ch s
CI Food Red 10, Azogeranine; CI (1975): 18050

INS: 128
2. nh ngha
Ch yu gm dinatri 8-acetamido-1-hydroxy-2-phenylazo3,6-naphthalendisulfonat cng cc cht mu ph cng vi

chnh.
Tn ha hc
Dinatri 8-acetamido-1-hydroxy-2-phenylazo-3,6naphthalendisulfonat
M s C.A.S.
3734-67-6
Cng thc ha hc
C18H13N3Na2O8S2
Cng thc cu to
OH
NHCOCH3
N
NaO3S
Khi lng phn t
509,43
3. Cm quan
Dng bt hoc ht mu .
4. Chc nng
Phm mu.
SO3Na
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 2,0%.
Cc cht hu c ngoi
Khng c qu 0,3% tng cc acid 5-acetamido-4hydroxy-2,7-naphthalen-2,7-disulfonic v acid 5-Amino-4-
cht mu
hydroxy-2,7-naphthalen-2,7-disulfonic.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Ch

Cc cht mu ph

Cc cht hu c ngoi
cht mu

2%/pht.
6.2. nh lng
Cn 0,6 - 0,7 g mu th;
mu (D).
Ph lc 12
YU CU K THUT V PHNG PHP TH I VI ALLURA RED AC
1. Tn khc, ch s
CI Food Red 17, FD&C Red No. 40; CI (1975): 16035

INS: 129
2. nh ngha
Ch yu gm dinatri 6-hydroxy-5-(2-methoxy-5-methyl-4sulfon-phenylazo)-2-naphthalen-sulfonat cng cc cht

phn khng mu chnh.
Tn ha hc
Dinatri 6-hydroxy-5-(2-methoxy-5-methyl-4-sulfonphenylazo)-2-naphthalen-sulfonat
M s C.A.S.
25956-17-6
Cng thc ha hc
C18H14N2Na2O8S2
Cng thc cu to
OCH3
NaO3S
HO
N
H3C
SO3Na
Khi lng phn t
496,43
3. Cm quan
Dng bt hoc ht mu sm.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 3,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,3% Natri 6-hydroxy-2-naphtalen

sulfonat;
Khng c qu 0,2% acid 4-amino-5-methoxy-2-methylbenzen-sulfonic;
Khng c qu 1,0% dinatri 6,6'-oxybis(2-naphthalensulfonat);

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1 tinh khit
Ch

Cc cht mu ph

Cc cht hu c ngoi
cht mu

- S dng k thut sc k lng di cc iu kin nh sau:
HPLC ra gii gradient t 0% n 18% vi tc tng c
nh 1%/pht; 18 n 62% vi tc tng c nh 7%/pht,
sau ra gii ti 100%, tc dng 0,6 ml/pht.
6.2. nh lng
Cn 0,5 - 0,6 g mu th;
mu (D).
Ph lc 13
YU CU K THUT V PHNG PHP TH I VI INDIGOTIN
1. Tn khc, ch s
Indigocarmin; CI Food Blue 1, FD&C Blue No. 2; CI (1975):

73015
INS: 132
2. nh ngha
Ch yu gm dinatri 3,3' -dioxo-[delta2,2'-biindolin]-5,5'disulfonat v dinatri 3,3'-dioxo-[delta2,2'-biindolin]-5,7'disulfonat cng cc cht mu ph cng vi NaCl v (hoc)
Na2SO4 l cc thnh phn khng mu chnh.
Tn ha hc
Dinatri 3,3' -dioxo-[delta2,2'-biindolin]-5,5'-disulfonat
M s C.A.S.
860-22-0 (ng phn 5,5')
Cng thc ha hc
C16H8N2Na2O8S2
Cng thc cu to
O
NaO3S
N
H
H
N
SO3Na
O
Khi lng phn t
466,36
3. Cm quan
Dng bt hoc ht mu xanh lam.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 1,0% (khng bao gm dinatri 3,3'-dioxo[delta2,2'-biindolin]-5,7'-disulfonat).
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% tng cc acid isatin-5-sulfonic; acid

5-sulfoanthranilic v acid anthranilic

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng

Khng c qu 18% dinatri 3,3'-dioxo-[delta2,2'-biindolin]5,7'-disulfonat.
6. Phng php th
6.1. tinh khit
Ch

Cc cht mu ph

s 3; Chiu cao tuyn dung mi ~ 17 cm.
Ch 1: ng phn 5,7' c tch ra l mt vng mu
xanh ln ngay trc vt chnh mu xanh. Khng tnh vng
ny vo khi xc nh cc cht mu ph.
Ch 2: 15 ml dung dch natri hydro carbonat s dng
trong quy trnh chung c thay th bng dung dch acid
hydrocloric 0,05 N trnh s phn hy xy ra khi indigo
sulfon ha trong mi trng kim.
Cc cht hu c ngoi
cht mu

- S dng k thut sc k lng vi cc iu kin nh sau:
HPLC ra gii gradient t 2 n 100%, sau tip tc ra
gii ti 100%.
Cc cht c th chit
bng ether

- Cn 2 g mu th thay cho hng dn cn 5 g mu trong
phn cc phng php chung.
6.2. nh lng
Cn 1,0 - 1,1 g mu th;
mu (D).
Xc nh ng phn bng sc k giy:
Tin hnh theo cc iu kin hng dn cho phn xc nh

hm lng cc cht mu ph. Ct vt sc k tng ng
ca ng phn trn sc k , theo hng dn ct vt mu
ph. Chit vo dung mi v o hp th quang ti Smax ca
n. Tin hnh o mu trng song song ti cng iu kin.
S dng 0,1 ml dung dch mu th lm chun, chm ln
bn giy hnh ch nht 18 cm x 0,7 cm.
Hm lng (%) ng phn trong mu c tnh nh sau
% = [A / As] x 20% x [D / 100]
Trong :
A l hp th ( hiu chnh vi mu trng) ca ng
phn.
As l hp th ( hiu chnh vi mu trng) ca chun.
D l hm lng tng cht mu trong mu.
Xc nh ng phn bng HPLC:
ng phn 5,7' c tch bng HPLC vi iu kin tch
cc cht mu ph nu trn v nh lng.
Ph lc 14
YU CU K THUT V PHNG PHP TH I VI BRILLIANT BLUE FCF
1. Tn khc, ch s
CI Food Blue 2, FD&C Blue No. 1; CI (1975): 42900

INS: 133
2. nh ngha
Ch yu gm dinatri 3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3sulfonbenzyl)-amino] phenyl] (2-sulfonphenyl)methylen]2,5-cyclohexadien-1-yliden] amonimethyl] benzensulfonat

v cc ng phn ca n cng cc cht mu ph cng vi
chnh.
Tn ha hc
Dinatri 3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3-sulfonbenzyl)amino] phenyl] (2-sulfonphenyl)methylen]-2,5cyclohexadien-1-yliden] amonimethyl] benzensulfonat.

Dinatri I-[4-(N-ethyl-3-sulfonatobenzylamino)phenyl]-I- [4(N-ethyl-3-sulfonatobenzyliminio)cyclohexa-2,5-dienyliden]toluen-2-sulfonat (Tn ha hc khc).
M s C.A.S.
3844-45-9
Cng thc ha hc
C37H34N2Na2O9S3
Cng thc cu to
CH2CH3
N CH2
SO 3
CH2
CH3
SO3Na
Khi lng phn t
792,86
3. Cm quan
Dng bt hoc ht mu xanh lam.
4. Chc nng
Phm mu.
SO 3Na
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Crom
Cc cht mu ph
Khng c qu 6,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 1,5% tng cc acid 2-; 3- v 4formylbenzensulfonic.

Khng c qu 0,3% acid 3-[[N-ethyl-N-(4-sulfophenyl)
amino] methyl] benzensulfonic.
Khng c qu 5,0%.
Leuco base

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Ch

Cc cht mu ph

s 4; Khai trin dung mi trong 20 gi.
Cc cht hu c ngoi
cht mu

- S dng k thut sc k ct vi cc iu kin hp th:
acid 3-formylbenzensulfonic: 0,0495 mgL-1cm-1 ti bc
sng 246 nm, trong HCl long.
acid
3-[[N-ethyl-N-(4-sulfophenyl)
amino]
methyl]
-1
-1
benzensulfonic: 0,078 mgL cm ti bc sng 277 nm,
trong dung dch amoniac long.
Leuco base
Th theo hng dn ti JECFA monograph 1-Vol. 4 - Cn

120 5 mg mu th v tin hnh theo hng dn trong
chuyn lun xc nh Leuco base trong cht mu nhm
Triarylmethan sulfon ha; hp th ring (a) = 0,164
mgL-1cm-1 ti bc sng ~ 630 nm; t s = 0,9706.
6.2. nh lng
Cn 1,8 - 1,9 g mu th;
mu (D).
Ph lc 15
YU CU K THUT V PHNG PHP TH I VI CHLOROPHYLL
1. Tn khc, ch s
Magnesium chlorophyll, Magnesium phaeophytin;

Clorophyllin; C.I. Natural Green 3; C.I. (1975): 75810;
INS 140
ADI "khng gii hn" .
2. nh ngha
Thu c bng cch chit c, c linh lng, tm ma v cc

nguyn liu thc vt khc vi dung mi; Sn phm sau khi
loi dung mi dng kt hp vi magnesi, c th loi
hon ton hoc 1 phn t clorophyll thu c
phaeophytin tng ng; Thnh phn mu chnh l cc
phaeophytin v magnesi clorophyll. Sn phm chit sau khi
loi dung mi cha cc cht mu khc nh carotenoid, sp
v cht bo c ngun gc t nguyn liu. Ch c s dng
nhng dung mi sau chit: aceton, dicloromethan,
methanol, ethanol, propan-2-ol v hexan.
Tn ha hc
Cc cht mu chnh l:
Phytyl (132R,17S,18S)-3-(8-ethyl-132-methoxycarbonyl2,7,12,18-tetramethyl-13'-oxo-3-vinyl-131-132-17,18tetrahydrocyclopenta [at]-prophylrin-17-yl) propionat
(Pheophytin a), hoc phc magnesi (Clorophyll a).
Phytyl (132R,17S,18S)-3-(8-ethyl-7-formyl-132methoxycarbonyl-2,12,18-trimethyl-13'-oxo-3-vinyl-131-13217,18-tetrahydro-cyclopenta [at]-prophylrin-17-yl) propionat,
(Pheophytin b), hoc phc magnesi (Clorophyll b).
M s C.A.S.
Phaeophytin a, phc magnesi: 479-61-8

Phaeophytin b, phc magnesi: 519-62-0
Phc magnesi phaeophytin a (Clorophyll a): C55H72MgN4O5
Phaeophytin a: C55H74N4O5
Phc magnesi phaeophytin b (Clorophyll a): C55H70MgN4O6
Phaeophytin b: C55H72N4O6
Cng thc ha hc
Cng thc cu to
Trong :
X = CH3 i vi cc hp cht "a"
X = CHO i vi cc hp cht "b".
Mt Mg s chuyn dng t clorophyll thnh phaeophytin
Khi lng phn t
Phc magnesi phaeophytin a (Clorophyll a): 893,51

Phaeophytin a: 871,22
Phc magnesi phaeophytin (Clorophyll a): 907,49
Phaeophytin : 885,20
3. Cm quan
Cht rn dng sp c mu t xanh oliu n xanh lc thm

ty thuc vo lng magnesi kt hp.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Sc k lp mng
Khng tan trong nc; tan trong cn, diethyl ether;

cloroalkan, hydrocarbon v du khng bay hi.
t yu cu (m t trong phn phng php th)
5.2. tinh khit

Dung mi tn d
Arsen
Aceton, methanol, ethanol, propan-2-ol, hexan: Khng c

qu 50,0 mg/kg, n cht hoc kt hp.
Dicloromethan: Khng c qu 10 mg/kg
(Th theo hng dn ti JECFA monograph 1-Vol. 4 Phng php II).
Ch

(Th theo hng dn ti JECFA monograph 1-Vol. 4. S
dng k thut hp th nguyn t thch hp xc nh. La
chn c mu th v phng php chun b mu da trn
nguyn tc ca phng php m t trong JECFA monograph
1-Vol. 4 phn cc phng php phn tch cng c).
5.3. Hm lng
Tng hm lng cc phaeophytin v cc phc magnesi ca

n khng thp hn 10,0%.
6. Phng php th
6.1. nh tnh
Sc k lp mng
Chm dung dch mu th 1/20 trong cloroform ln bn mng

Silica 60C thnh 1 vch di 2 cm. Sau khi bn mng kh,
khai trin bn mng trong h dung mi gm 50% hexan, 45%
cloroform v 5% ethanol (thng thng cloroform vi tinh
khit thuc th cha sn 2% ethanol lm n nh.
T l 5% ethanol l khng tnh n lng ethanol c sn

ny) n khi tuyn dung mi t 15 cm cch im xut pht.
dung mi bay hi, sau quan st cc vt ring bit v
xc nh vt tng ng vi cu t quan tm thng qua Rf v
mu. Gi tr Rf gn ng v mu ca cc vt nh sau:
phaeophytin a: 0,77; mu xm/nu
phaeophytin b: 0,75; mu vng/nu
Clorophyll a: 0,50; mu xanh lam/xanh lc
Clorophyll b: 0,63; mu vng/xanh lc
Ngoi ra cn quan st thy cc vt ca -caroten ti Rf 0,81
v xanthophyll ti Rf 0,47 v 0,23.
6.2. tinh khit
Dung mi tn d
Th theo hng dn ti JECFA monograph 1-Vol. 4.

Xc nh bnh Sc k kh hoc bng phng php ct cun
(chuyn lun Xc nh dung mi tn d) hoc phn tch
khng gian hi (chuyn lun Th gii hn dung mi tn d).
Arsen

- Phng php II.
Ch

- S dng k thut hp th nguyn t thch hp xc nh.
La chn c mu th v phng php chun b mu da
trn nguyn tc ca phng php m t trong JECFA
monograph 1-Vol. 4 phn cc phng php phn tch cng
c).
6.1. nh lng
Cn khong 100 mg (chnh xc n mg) mu th, ha tan

trong diethyl ether n 100 ml. Ly 2 ml dung dch ny,
pha long vi diethyl ether va 25 ml. hp th quang
ca mu ti nng ny khi o ti bc sng 660,4 nm
khng c vt qu 0,7 so vi thang o hp th.
o hp th quang (A) ca dung dch ny trong cuvet 1 cm
so vi mu trng l diethyl ether ti bc sng 660,4 nm;
642,0 nm; 667,2 nm; 654,4 nm. (cc bc sng ny ln lt
l cc i hp th trong diethyl ether ca clorphyll a;
clorophyll b; phaeophtin a v ng phaeophytin b, tng
ng). Ngoi ra o thm ti bc sng 649,8 nm v 628,2 nm.
Phn dung dch pha long cn li, thm vo 1 tinh th
acid oxalic, ha tan v lc u. o li hp th quang ca
dung dch ti cc bc sng cho. " A" l s chnh lch
hp th quang ca dung dch o ti cng bc sng trc
v sau khi thm acid oxalic.
Tnh nng tng cht ( mol/L) theo cc phng trnh sau:
Clorophyll a = 17,7 A (660,4 nm) + 7,15 A (642,0 nm)
Clorophyll b = 19,4 A (642,0 nm) - 2,92 A (660,4 nm)
Phaeophytin a = -4,89 A (649,8 nm) + 0,0549 A (628,2 nm)
+18,7 A (667,2 nm) + 0,0575 A (654,4 nm) - clorophyll a
Phaeophytin b = -71,0 A (649,8 nm) + 2,51 A (628,2 nm) 13,5 A (667,2 nm) + 84,3 A (654,4 nm) - clorophyll b
Chuyn t mol/L sang hm lng % theo phng trnh sau:
% clorophyll a =a
% clorophyll b =a
% phaeophytin a =a
% phaeophytin b =a
mol 0,8935 12,5 100

Khi lng mu th (mg)
mol 0,9075 12,5 100
Khi lng mu th (mg)
mol 0,8712 12,5 100
Khi lng mu th (mg)
mol 0,8852 12,5 100
Khi lng mu th (mg)
Ph lc 16
YU CU K THUT V PHNG PHP TH I VI
CHLOROPHYLL PHC NG
1. Tn khc, ch s
Copper chlorophyllin, copper phaeophytin

C.I. Natural Green 3; C.I. (1975) No. 75810;
INS: 141i
2. nh ngha
Thu c khi thm mui ng hu c vo sn phm thu

c khi chit c, c linh lng, tm ma v cc nguyn liu
thc vt khc vi dung mi; Sn phm sau khi loi dung
mi cha cc cht mu khc nh carotenoid cng nh sp
v cht bo. Cht mu chnh l ng phaeophytin. Ch c
s dng nhng dung mi sau chit: aceton,
dicloromethan, methanol, ethanol, propan-2-ol v hexan.
Tn ha hc
Cht mu chnh dng acid ca n l

[Phytyl (132R, 17S, 18S)-3-(8-ethyl-132-methoxycarbonyl2,7,12,18-tetramethyl-13-oxo-3-vinyl-131, 132,17,18tetrahydroxypenta[at]-prophyrin-17-yl) propionat] ng II
(ng phaeophytin a)
[Phytyl (132R, 17S, 18S)-3-(8-ethyl-7-formyl-132methoxycarbonyl-2,12,18-trimethyl-13-oxo-3-vinyl-131,
132,17,18-tetrahydroxypenta[at]-prophyrin-17-yl) propionat]
ng II (ng phaeophytin b).
M s C.A.S
65963-40-8
Cng thc ha hc
ng phaeophytin a (dng acid): C55H72CuN4O5

ng phaeophytin b (dng acid): C55H70CuN4O6
Cng thc cu to
Vi:
X = CH3 i vi hp cht "a"
X = CHO i vi hp cht "b"
Khi lng phn t
ng phaeophytin a: 932,75
ng phaeophytin b: 943,73
3. Cm quan
Cht rn dng sp c mu t xanh lc n xanh lc thm ty

thuc vo ngun nguyn liu.
4. Chc nng
Phm mu.
5. Tnh cht
5.1. nh tnh
tan
Quang ph
Sc k lp mng
Khng tan trong nc; tan trong cn, diethyl ether;

cloroalkan, hydrocarbon v du khng bay hi.
A (1%, 1 cm) ca mu th trong cloroform, o ti 405 nm
khng c thp hn 54.
t yu cu (m t trong phn phng php th).
5.2. tinh khit

Dung mi tn d
ng t do c th ion
ha

qu 50 mg/kg, n cht hoc hp cht.
Dicloromethan: Khng c qu 10 mg/kg.
Khng c qu 200 mg/kg.
ng tng s
Khng c qu 8% tnh theo tng s ng phaeophytin.
Arsen
Khng c qu 3 mg/kg.
Ch
Khng c qu 5 mg/kg.
5.3. Hm lng
Khng thp hn 10% tng s ng phaeaphytin.
6. Phng php th
6.1. nh tnh
Sc k lp mng
Chm dung dch mu th 1/20 trong cloroform ln bn mng

Silica 60C thnh 1 vch di 2 cm. Sau khi bn mng kh,
khai trin bn mng trong h dung mi gm 50% hexan, 45%
cloroform v 5% ethanol (thng thng cloroform vi tinh
khit thuc th cha sn 2% ethanol lm n nh. 5%
ethanol l cha tnh n lng ethanol c sn ny) n khi
tuyn dung mi t 15 cm cch im xut pht. dung mi
bay hi, sau quan st cc vt ring bit v xc nh vt
tng ng vi cu t quan tm thng qua Rf v mu. Gi tr
Rf gn ng v mu ca cc vt nh sau:
ng phaeophytin a: 0,5; mu xanh lc
ng phaeophytin b: 0,73; mu vng/xanh lc
Ngoi ra cn quan st thy cc vt ca -caroten ti Rf 0,81
v xanthophyll ti Rf 0,47 v 0,23.
6.2. tinh khit

Dung mi tn d

- Xc nh bnh Sc k kh hoc bng phng php ct cun
ng t do c th ion
ha
Cn khong 1 g (chnh xc n mg) mu th v ha tan

trong 20 ml du lc, un nng nh cho tan hon ton. Thm
chnh xc 200 ml nc, khuy bng my khuy t, v iu
chnh ti pH 3,0 bng cch thm cn thn acid hydrocloric
0,5 N (trnh thm qu nhanh). yn hn hp trong 10 pht.
Nu cn, iu chnh li ti pH 3,0 bng cch thm cn thn
acid hydrocloric 0,5 N. Chuyn sang phu tch v yn 20
pht. Lc lp nc qua giy lc Whatman No. 50, b 10 ml
dch lc u. Dng dung dch ny phn tch ng bng
quang ph hp th nguyn t - JECFA monograph 1-Vol. 4.
ng tng s
Cn khong 0,1 g (chnh xc n mg) mu th, cho vo a

silica, nung nhit khng qu 500 C, ti khi loi i ton
b than; lm m tro bng mt hoc hai git acid sulfuric c
v tro ha li. Ha tan tro bng cch un si 3 ln, mi ln
vi 5 ml acid hydrocloric 10% (kl/kl), lc dch acid thu c
sau mi ln thm acid qua cng mt phu giy lc nh vo
bnh nh mc 100 ml. ngui, v thm nc tinh khit ti
vch. Dng dung dch ny phn tch ng bng quang
ph hp th nguyn t - xem JECFA monograph 1-Vol. 4.
Arsen
- Th theo hng dn ti JECFA monograph 1 - Vol.4

- Phng php II.
Ch

- S dng k thut hp th nguyn t thch hp xc nh.
La chn c mu th v phng php chun b mu da
trn nguyn tc ca phng php m t trong JECFA
monograph 1-Vol. 4 phn cc phng php phn tch cng
c.
6.3. nh lng
Cn khong 100 mg (chnh xc n mg) mu th, ha tan

trong diethyl ether n 100 ml. Ly 2 ml dung dch ny,
pha long vi diethyl ether va 25 ml. hp th quang
ca mu ti nng ny khi o ti bc sng 660,4 nm
khng c vt qu 0,7 so vi thang o hp th.
o hp th quang (A) ca dung dch ny trong cuvet 1 cm
so vi mu trng l diethyl ether ti bc sng 667,2 nm;
654,4 nm; 649,8 nm v 628,2 nm. (hai bc sng sau l cc
i hp th trong diethyl ether ca ng phaeophtin a v
ng phaeophytin b, tng ng).
Tnh nng tng cht ( mol/L) theo cc phng trnh sau:

ng phaeophytin a = 45,6 A (649,8 nm) - 2,75 A (628,2 nm)
+ 3,10 A (667,2 nm) - 35,4 A (654,4 nm)
ng phaeophytin b = -8,46 A (649,8 nm) + 20,7 A (628,2
nm) - 1,69 A (667,2 nm) + 5,13 A (654,4 nm)
Chuyn t mol/L sang hm lng % theo phng trnh sau:
mol 0,9327 12,5 100
% ng phaeophytin a =
Khi lng mu th (mg)
% ng phaeophytin b =
mol 0,9467 12,5 100

Khi lng mu th (mg)
Ph lc 17
YU CU K THUT V PHNG PHP TH I VI MUI NATRI, KALI CA
CLOROPHYL PHC NG
1. Tn khc, ch s
2. nh ngha
Natri ng clorophyllin; kali ng clorophyllin

Chlorophyllins, C.I. (1975) No. 75810;
INS 141ii
ADI = 0 -15 mg/kg th trng.
Mui kim ca ng Clorophyllin thu c khi thm ng
vo sn phm thu c khi x phng ha dch chit dung
mi t c, c linh lng, tm ma v cc nguyn liu thc vt
khc; qu trnh x phng ha c th loi cc nhm ester
methyl v cyclophytol v c th tch i mt phn vng
pentenyl; sau khi thm ng vo clorophyllin tinh ch,
nhm acid b trung tnh ha to thnh dng mui ca kali
v/hoc natri; sn phm thng mi c th dng dung
dch nc hoc bt kh. Ch c s dng nhng dung mi
sau chit: aceton, dicloromethan, methanol, ethanol,
propan-2-ol v hexan.
Tn ha hc
Cht mu chnh dng acid ca n l

3-(10-Carboxylato-4-ethyl-1,3,5,8-tetramethyl-9-oxo-2vinylphorbin-7-yl) propionat, phc ng (ng clorophyllin a)
3-(10-carboxylato-4-ethyl-3-formyl-1,5,8-trimethyl-9-oxo-2vinylphorbin-7-yl) propionat, phc ng (ng clorophyllin b)
Ty thuc vo mc thy phn vng cyclopentenyl c th
tch i vi sn phm thu c vi chc carboxyl th ba.
Cng thc ha hc
ng clorophyllin a (dng acid): C34H32CuN4O5

ng clorophyllin b (dng acid): C34H30CuN4O6
Cng thc cu to
Vi:
X = CH3 i vi hp cht "a"
X = CHO i vi hp cht "b"
M = Kali v/hoc natri
Khi lng phn t
ng clorophyllin a: 640,20
ng clorophyllin b: 654,18
Mi cht c th tng ti 18 Dalton nu vng cyclopentenyl
tch i.
3. Cm quan
Dung dch mu l cy sm hoc bt xanh l cy sm ti

xanh lam/en.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc; rt kh tan trong cn thp , keton v

diethyl ether; khng tan trong cloroalkan, hydrocarbon v
mt s du khng bay hi.
Quang ph
ng
Natri
Phi c phn ng c trng ca natri.
Kali
Phi c phn ng c trng ca kali.
5.2. tinh khit

Phm mu kim
Dung mi tn d

qu 50,0 mg/kg, n cht hoc kt hp.
Dicloromethan: Khng c qu 10,0 mg/kg
ng t do c th ion
ha
ng tng s
Khng c qu 8,0% tnh theo ng phaeophytin tng s.
Arsen
Ch
5.3. Hm lng
Khng thp hn 95,0% ng Clorophyllin tng s tnh theo

ch phm sy kh ti 100 C trong 1 gi.
6. Phng php th
6.1 nh tnh
Quang ph
A (1%, 1 cm) ca mu th ( sy kh 100 C trong 1 gi),

o ti 405 nm trong dung dch m phosphat pH 7,5 khng
c nh hn 540.
ng
Natri
Kali
Ha tan tro sulfat ca mu th (Cn 1 g mu, Phng php

I) trong 10 ml acid hydrocloric long (TS) bng cch lm
nng trong b cch thy. Lc nu dung dch c, v pha
long vi nc ti 10 ml. S dng dung dch ny l dung
dch th cho cc php th sau:
Cho dung dch amoniac (TS) vo 5 ml dung dch th kim
ha. Dung dch phi xut hin mu xanh lam.
Cho 0,5 ml dung dch natri diethyldithiocarbamat 1/1000 vo
5 ml dung dch th. Dung dch phi xut hin ta nu.
Th theo hng dn ti JECFA monograph 1-Vol. 4 - Chun
b dung dch c m t trong chuyn lun nh tnh ng.
Th theo hng dn ti JECFA monograph 1-Vol. 4 - Chun
b dung dch c m t trong chuyn lun nh tnh ng.
6.2 tinh khit

Phm mu kim
Ly 5 ml dung dch mu th 0,5% trong nc vo ng

nghim, thm 1 ml acid hydrocloric 1 N v 5 ml diethyl ether.
Lc k v phn lp. Mu ca lp ether khng c m
hn mu xanh l cy nht.
Dung mi tn d
Xc nh bnh Sc k kh hoc bng phng php ct cun

(Th theo hng dn ti JECFA monograph 1-Vol. 4)
ng t do c th ion
ha
Cn khong 1 g (chnh xc n mg) mu th v ha tan

trong 20 ml du lc, un nng nh. Thm chnh xc 200 ml
nc, khuy bng my khuy t, v iu chnh ti pH 3,0
bng cch thm cn thn acid hydrocloric 0,5 N (trnh thm
qu nhanh). yn hn hp trong 10 pht. Nu cn, iu
chnh li ti pH 3,0 bng cch thm cn thn acid hydrocloric
0,5 N. Chuyn sang phu tch v yn 20 pht. Lc lp
nc qua giy lc Whatman No. 50, b 10 ml dch lc u.
Dng dung dch ny phn tch ng bng quang ph hp
th nguyn t (xem JECFA monograph 1-Vol. 4).
ng tng s
Cn khong 0,1 g (chnh xc n mg) mu th, cho vo a

silica, nung nhit khng qu 500 C, ti khi loi i ton
b than; lm m tro bng mt hoc hai git acid sulfuric c
v tro ha li. Ha tan tro bng cch un si 3 ln, mi ln
vi 5 ml acid hydrocloric 10% (kl/kl), lc dch acid thu c
sau mi ln thm acid qua cng mt phu giy lc nh vo
bnh nh mc 100 ml. ngui, v thm nc tinh khit ti
vch. Dng dung dch ny phn tch ng bng quang
ph hp th nguyn t (xem JECFA monograph 1-Vol. 4).
Arsen
Th theo hng dn ti JECFA monograph 1-Vol. 4Phng php II
Ch

dng k thut hp th nguyn t thch hp xc nh. La
nguyn tc ca phng php m t trong JECFA

monograph 1-Vol. 4 phn cc phng php phn tch
cng c.
6.3. nh lng
Cn khong 1 g (chnh xc n mg) mu th, sy kh
trc 100 C trong 1 gi, sau ha tan trong 20 ml dung
dch m phosphat (pH 7,5) v pha long ti 1000 ml bng
nc ct. Ly 10 ml dung dch ny v pha long ti 100 ml
bng dung dch m phosphat (pH 7,5). o mt quang
ca dung dch ny (0,001% kl/tt) bng my quang ph thch
hp, s dng cuvet o 1 cm v rng khe 0,10 mm 403406 nm, ghi cc i trong khong .
Hm lng (%) natri ng clorophyllin c tnh theo cng
thc theo cng thc sau:
Mt quang 104
-------------------------------------565 khi lng mu th (g)
Quy c: natri ng clorophyllin tinh khit 100% c hp
th ring l 565.
Ph lc 18
YU CU K THUT V PHNG PHP TH I VI GREEN S
1. Tn khc, ch s
CI food green 4, food green S; CI (1975): 44090

INS: 142
ADI khng xc nh.
2. nh ngha
Ch yu gm natri N-[4-][[4-(dimethylamino) phenyl] (2hydroxy-3,6 disulfo-1-naphtalenyl) methylen]-2,5cyclohexadien-1-ylidenen-N-methylmetanamini v cc cht

phn khng mu chnh.
Tn ha hc
Natri N-[4-[[4-(dimethylamino) phenyl](2-hydroxy-3,6

disulfo-1- naphtalenyl) methylen]-2,5-cyclohexadien-1yliden]-N methylmetanamini;
hoc Natri 5-[4-dimethylamino-l-(4dimethyliminiocyclohexa-2,5-dienyliden) benzyl]-6-hydroxy7-sulfonat-naphtalen-2-sulfonat.
M s C.A.S.
860-22-0
Cng thc ha hc
C27H25N2NaO7S2
Cng thc cu to
N(CH3)2
-
O 3S
OH
C
NaO 3S
N(CH3)2
Khi lng phn t
576,63
3. Cm quan
Dng bt hoc ht mu xanh lc m.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, kh tan trong ethanol.
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC
Khng c qu 20% cng vi clorid v sulfat tnh theo

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Khng c qu 2 mg/kg.
Crom
Khng c qu 50 mg/kg.
Cc cht mu ph
Khng c qu 1%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,1% ru 4, 4-bis (dimethylamino)

benzhydric.
Khng c qu 0,1% 4, 4-bis (dimethylamino)
benzophenon,
Khng c qu 0,2% acid 3-Hydroxynaphtalen-2,7disulfonic.

khng sulfon ho
Leuco base
Khng c qu 5,0%.
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
Khng c thp hn 80% tng cc cht mu.
6. Phng php th
6.1. tinh khit
Ch

Cc cht mu ph

- S dng iu kin nh sau: Dung mi khai trin l dung
mi s 2, chiu cao t vch xut pht ti tuyn dung mi
khong 17 cm.
Cc cht hu c ngoi
cht mu

- S dng k thut sc k lng vi iu kin nh sau:
HPLC ra gii gradient 2% n 100% vi tc tng c
nh 2%/pht.
Leuco base

- Cn chnh xc 110 5 mg mu v tin hnh theo hng
dn trong chuyn lun th Leuco base trong cht mu
Triarylmethan sulfon ha. hp th ring (a)=0,1725
mgL-1cm-1 ti bc sng khong 634 nm. T s = 0,9600.
6.2. nh lng
Cn 1,4 - 1,5 g mu th;
mu (D).
Ph lc 19
YU CU K THUT V PHNG PHP TH I VI CARAMEN
1. Tn khc, ch s
2. nh ngha
3. Cm quan
4. Chc nng
Cht mu caramel c chia thnh bn loi. Tn khc v

ch s ca mi loi nh sau:
Loi I: Caramel thng, caramel caustic; INS No. 150a
Loi III: Caramel amoni; INS No. 150c
Loi IV: Caramel amoni sulfit, INS No. 150d
ADI i vi nhm I Khng xc nh;
ADI i vi nhm III = 0-160 mg/kg th trng
ADI i vi loi IV 0-200 mg/kg th trng (0-150 mg/kg th
trng i vi dng rn)
L cc hn hp phc tp, trong mt s dng t hp
keo, sn xut bng cch un nng ring carbohydrat hoc
cng vi s c mt ca acid, kim hoc mui loi thc
phm; ty theo ha cht dng khi sn xut phn loi nh
sau:
Loi I: sn xut bng cch un nng ring carbohydrat
hoc cng vi s c mt ca acid, kim; khng dng hp
cht amoni hoc sulfit.
Loi III: sn xut bng cch un nng ring carbohydrat
hoc cng vi s c mt ca acid, kim; cng vi hp cht
amoni; khng dng hp cht sulfit.
Loi IV: sn xut bng cch un nng ring carbohydrat
hoc cng vi s c mt ca acid, kim; cng vi c hp
cht sulfit v hp cht amoni.
Trong tt c cc trng hp, nguyn liu carbohydrat c
sn dng thng phm l cc cht lm ngt dinh dng
loi thc phm c cha glucose, fructose v/hoc polyme
ca chng. Cc acid v kim l acid sulfuric hoc citric thc
phm v natri, kali hoc calci hydroxyd hoc hn hp ca
chng loi thc phm.
Hp cht amoni c dng l mt hoc bt k mt cht no
sau y: amoni hydroxyd, amoni carbonat v amoni hydro
carbonat, amoni phosphat, amoni sulfat, amoni sulfit v
amoni hydrosulfit.
Dng hp cht sulfit l mt hoc bt k mt cht no sau
y: acid sulfur, kali, natri v amoni sulfit v amoni hydro
sulfit.
C th dng tc nhn chng ni bt loi thc phm tr
gip qu trnh sn xut.
Cht dng rn hoc lng, mu nu sm ti en c mi
ng chy.
Phm mu
5. Yu cu k thut
5.1. nh tnh
tan
C th trn ln vi nc
nh tnh tp mu
Phi c phn ng c trng ca tp mu.
Phn loi
Loi I: Khng c qu 50 % cht mu l loi lin kt bi

celulose DEAE v khng qu 50% cht mu l loi lin kt
bi celulose phosphoryl.
Loi III: Khng c qu 50% cht mu l loi lin kt bi
celulose DEAE v hn 50% cht mu l loi lin kt bi
celulose phosphoryl.
Loi IV: Hn 50% cht mu l loi lin kt bi celulose
DEAE v t s hp th khng qu 50.
Xem m t trong phn PHNG PHP TH.
Ch : Gii hn arsen v ch p dng cho tt c cc nhm
caramel v c tnh da trn ch phm nguyn dng:
Cc gii hn v khong khc p dng i vi mi loi ring
theo ch dn, nu khng c ch dn ring, c tnh da
trn ch phm rn.
Loi I: 62,0 77,0%
Loi III: 53,0 83,0%
Loi IV: 40,0 75,0%
Loi I: 0,01 - 0,12
Loi III: 0,08 - 0,36
Loi IV: 0,10 - 0,60
Loi I: Khng c qu 0,1%
Loi III: 1,3 - 6,8%
Loi IV: 0,5 - 7,5%
Loi I: Khng c qu 0,3%
Loi III: Khng c qu 0,3%
Loi IV: 1,4B- 10,0%
Loi I: Loi III: Loi IV: Khng c qu 0,5%
Loi I: Loi III: Khng c qu 0,4%
Loi IV: Khng c qu 2,8%
Loi I: Loi III: Khng c qu 300,0 mg/kg & khng c qu
200,0 mg/kg tnh theo cht mu tng ng
Loi IV: Khng c qu 1000,0 mg/kg & khng c qu
250,0 mg/kg tnh theo cht mu tng ng
5.2. tinh khit
Hm lng cht rn
Cng mu
Nit ton phn
Lu hunh ton phn
Lu hunh dioxyd
Nit amoniac
4-Methylimidazol (MEI)
2-Acetyl-4-tetrahydroxy- Loi I: butylimidazol (THI)

Loi III: Khng c qu 40,0 mg/kg & khng c qu
25,0 mg/kg tnh theo cht mu tng ng.
Loi IV: Arsen
Khng c qu 1,0 mg/kg
Ch
Khng c qu 2,0 mg/kg
6. Phng php th
6.1. nh tnh
Phn loi/ Cht mu lin Trong ch tiu ny cht mu lin kt bi celulose DEAE
kt bi celulose DEAE
c nh ngha l phn trm hp th gim i ca dung
dch cht mu caramel 560 nm sau khi x l vi celulose
DEAE.
Thuc th ring :
Celulose DEAE (diethylaminoethyl) c dung lng 0,7
mE/g, nh Cellex D ca Bio-Rad hoc Celulose DEAE c
dung lng cao hn hoc thp hn tng ng vi lng
cht cao hn hoc thp hn.
Tin hnh:
Pha dung dch cht mu caramel c hp th xp x 0,5
ti 560 nm bng cch cho mt lng cht mu caramel
thch hp vo bnh nh mc 100 ml, cng vi acid
hydrocloric 0,025 N. Pha long ti vch bng acid
hydrocloric 0,025 N v ly tm hoc lc, nu dung dch vn
c. Ly 20 ml dung dch cht mu caramel, thm 200 mg
celulose DEAE, trn k trong t pht, ly tm hoc lc, v thu
ly lp dung dch trong pha trn. Xc nh hp th ca
dung dch cht mu caramel v lp dung dch pha trn
trong cng o 1 cm 560 nm, bng my quang ph thch
hp, ngay trc chun ha vi mu trng l acid
hydrocloric 0,025 N. Tnh phn trm cht mu lin kt bi
celulose DEAE theo cng thc:
(A 1 A 2 )
10 0
A1
Trong :
A1 l hp th ca dung dch cht mu caramel 560 nm
A2 l hp th ca lp dung dch pha trn sau khi x l
vi Celulose DEAE 560 nm.
Phn loi/ Cht mu lin Trong ch tiu ny cht mu lin kt bi celulose
kt bi celulose
phosphoryl c nh ngha l phn trm hp th gim
phosphoryl
i ca dung dch cht mu caramel 560 nm sau khi x l
vi celulose phosphoryl.
Thuc th ring:
Celulose phosphoryl dung lng 0,85 mE/g, nh Cellex P
Bio-Rad hoc Celulose phosphoryl dung lng cao hn
hoc thp hn tng ng vi lng cht cao hn hoc
thp hn.
Tin hnh:
Cho khong 200-300 mg cht mu caramel vo bnh nh
mc 100 ml, pha long ti vch bng acid hydrocloric
0,025 N v ly tm hoc lc, nu dung dch vn c. Ly 40
ml dung dch cht mu caramel, thm 2,0 g celulose
phosphoryl, lc k trong t pht. Ly tm hoc lc, v thu ly
lp dung dch trong pha trn. Xc nh hp th ca
dung dch cht mu caramel v lp dung dch pha trn
trong cng o 1 cm 560 nm, bng my quang ph thch
hp, ngay trc chun ha vi mu trng l acid
hydrocloric 0,025 N. Tnh phn trm cht mu lin kt bi
celulose phosphoryl theo cng thc:
(A 1 A 2 )
10 0
A1
Trong :
A1 l hp th ca dung dch cht mu caramel 560 nm
A2 l hp th ca lp dung dch pha trn sau khi x l
vi Celulose phosphoryl 560 nm.
Phn loi/ H s hp Trong ch tiu ny t s hp th c nh ngha l
th
hp th ca cht mu caramel 280 nm chia cho hp
th ca cht mu caramel 560 nm.
Tin hnh:
Cho khong 100 mg cht mu caramel vo bnh nh mc
100 ml cng vi nc, pha long ti vch bng nc, lc
u v ly tm, nu dung dch vn c. Ly 5,0 ml dung dch
trong ny cho vo bnh nh mc 100 ml, pha long ti
vch bng nc, v lc u. Xc nh hp th ca dung
dch 0,1% trong cng o 1 cm 560 nm v ca dung dch
pha long 1:20 280 nm, bng my quang ph thch hp,
ngay trc chun ha vi mu trng l nc. (My
quang ph thch hp l my c b n sc ha cho
rng di 2 nm hoc nh hn v t l nh sng lc l 0,5%
hoc nh hn.) Tnh t s hp th ca cht mu
caramel bng cch chia gi tr hp th ca cht mu
caramel 280 nm nhn 20 (h s pha long) cho gi tr
hp th 560 nm.
6.2. tinh khit
Hm lng cht rn
Hm lng cht rn ca cht mu Caramel c xc nh

bng cch lm kh mu th vi cht mang gm c ct
thch anh tinh khit qua c ry s 40 nhng khng qua
c c ry s 60 c chun b bng cch ra vi
acid hydrocloric, ra ht acid, sy v nung. Trn 30,0 g ct
chun b c cn chnh xc vi 1,5-2,0 g cht mu
caramel c cn chnh xc v lm kh ti khi lng
khng i 60oC di p sut gim 50 mmHg (6,7 kPa).
Ghi khi lng cui cng ca caramel v ct. Tnh % cht
rn theo cng thc sau:
% cht rn =
(m 3
m2 )
10 0
m1
Trong :
m3 = khi lng cui cng ca caramel thm ct (g)
m2 = khi lng ct (g)
m1 = khi lng caramel thm vo ban u (g)
Tnh ton trn c s cht rn
Hm lng nit ton phn, lu hunh ton phn,
amoni, lu hunh dioxyd, 4-MEI v THI c biu th
cht rn. Xc nh nng (Ci) ca mi tp cht
nguyn trng; nng (CS) theo cht rn c tnh
cng thc:
Cs = (Ci
Cng mu
nit
theo
theo
theo
100) / % cht rn
Trong ch tiu ny cng mu c nh ngha l

hp th ca dung dch 0,1% (kl/tt) ca cht mu caramel
rn trong nc 610 nm trong cng o 1 cm.
Tin hnh:
Cho khong 100 mg cht mu caramel vo bnh nh mc
100 ml cng nc, pha long ti vch bng nc, lc u
v ly tm, nu dung dch vn c. Xc nh hp th
(A610) ca dung dch trong ny trong cng o 1 cm 610
nm bng my quang ph thch hp, ngay trc
chun ha vi mu trng l nc. Tnh cng mu ca
cht mu caramel theo cng thc sau:
Cng mu = (A610
100) / % cht rn
Xc nh % cht rn nh m t phn Hm lng cht

rn.
Tnh ton trn c s cht mu tng ng: Khi c thm
cc gii hn i vi 4-MEI v THI c biu th theo cht
mu tng ng th cc nng trc ht c tnh trn
c s cht rn nh ch dn trong phn "Tnh trn c s vo
cht rn", v sau biu th theo cht mu tng ng
theo cng thc:
CS 0,1
Mu tng ng =
cng mu
Trong :
CS = nng trn cht rn.
Nh vy hm lng c biu th bng mt sn phm c
cng mu l 0,1 n v hp th.
Nit ton phn
Lu hunh ton phn
Lu hunh dioxyd
Th theo hng dn ti JECFA monograph 1-Vol. 4- Xc

nh theo ch dn trong phn Xc nh Nit (Phng php
Kjeldahl) s dng Phng php II
Ly mt chn ln nht c th cho va l nung in, cho
vo 1-3 g MgO hoc mt lng tng ng
Mg(NO3)26H2O (6,4 19,2 g), 1 g ng saccarose
lm thnh bt, v 50 ml HNO3. Thm 5-10 g cht mu
caramel. Cho cng mt lng thuc th vo mt chn khc
lm mu trng. C trn hi nc si ti khi thnh khi
nho. t chn vo l nung ngui (25 C) v un nng t
t ti khi ton b khi NO2 bay ht. ngui, ha tan v
trung tnh ha bng HCl (1+2,5), thm d 5 ml. Lc, un
nng ti si, v thm 5 ml dung dch BaCl22H2O 10% tng
git mt. C ti 100 ml, yn qua m, lc, ra, nung, v
cn BaSO4. Hiu chnh kt qu i vi BaSO4 thu c
trong mu trng v ghi dng mg S/100 g. Cc dng c
thng mi dng phn tch lu hunh ton phn nh
thit b phn tch theo qui trnh t chy Leco/chun
cng c th s dng v c khuyn dng vi lng mu
th khong 200 mg.
Thit b
S dng thit b Monier-Williams ci tin (c 5GA
Scientific, Inc., Bloomfield, N.J., M) xc nh acid
sulfur, hoc dng c thit k nh trnh by hnh. B
dng c gm mt bnh ct y trn ba c 1000 ml c ng
ni thy tinh vut thon chun 24/40. Gn km mt sinh hn
Allihn 30 cm hi lu mt c pha ngoi ca bnh, v u
kia ca sinh hn ni vi mt ng Tygon hoc silicon inch
(c un si trc vi dung dch acid hydrocloric 1/20 v
ra bng nc) ti b ng hp th (c ni hnh cu 35/20
hoc tng ng). Ni c gia ca bnh vi bnh gn
hnh tr 125 ml, v gn mt on ng ti ng ch U ngn
cm qua mt np cao su trn c ca bnh gn. Gn mt
ng vo bng thy tinh, cong di, ti gn y bnh, vo c
ngoi khc ca bnh, v ni ng vo mt bnh ra-kh 250
ml mt on ng. Bnh ra kh li ni bng mt ng ti chai
(bom kh) nit.
Nghin 4,5 g pyrogalol (acid pyrogalic) vi 5 ml nc trong

mt ci nh, v cho khi nho sang bnh ra kh. Tip tc
nghin phn cn li, v chuyn ton b sang bnh vi 2 ln
5 ml nc. Cho kh nit i t chai kh ti bnh ui ht
khng kh, v sau thm vo bnh qua phu cung di

dung dch c lm ngui ca 65 g kali hydroxyd trong
khong 85 ml nc. Lp u bnh vo, v sc bong bng
nit qua ui ht khng kh khi khng gian pha
trn. Kp cht ng c hai v tr ca chai, v ni n ti ng
u vo ca bnh ct. Bnh ra kh phi c chun b nh
m t, vi dung dch pyrogalol mi pha trong ngy lm.
Thm ln lt vo mi ng hp th hnh ch U nh sau:
hai a thy tinh 8 mm chiu di khong 25 mm, 10 ml ht
thy tinh 3 mm u ra, 10,0 ml dung dch hydrogen
peroxyd 3%, v 1 git methyl (TS).
Lp tt c cc phn ca thit b, v kim tra xem c kn
khng bng cch thi nh vo trong ng gn vi c bnh
gn. Khi thi, ng kha bnh gn. yn vi pht; nu
mc cht lng trong ng ch U vn ngang bng, ng tt
c cc khp ni v th li. Nu h thng kht th tin hnh
theo ch dn di y.
Tin hnh:
Phn tn khong 25 g mu th, cn chnh xc, trong 300
ml nc va un si ngui, v chuyn khi nho sang
bnh qua mt phu l rng dng nc. Pha long ti
khong 400 ml bng nc, v kha bnh gn li. Thm 90
ml acid hydrocloric 4 N vo bnh gn, v cho acid vo bnh
bng cch thi nh vo ng c bnh gn. Kha bnh gn.
M kp ng c hai v tr ca bnh ra kh, v bt u thi
kh nit vi tc u n bt kh. un nng bnh ct bng
bp o un hi lu trong khong 20 pht. Khi t c
s hi lu n nh, cp in un tip hi lu trong 1,75 gi.
Tt nc trong sinh hn, v tip tc un ti khi khp ni
pha u vo ca ng ch U th nht xut hin hi ngng
ng v hi m. B bnh gn v tt bp.
Khi khp ni u trn ca sinh hn ngui, tho khp ni v
trng vo ng ch U th hai, li ng ni vi khp ra ca
ng ch U th nht nhng ngt khi u vo ca ng ch
U th hai. Quay ng ni ti khi u h ca n gn chm
u vo ca ng ch U th nht. Thm 1 git methyl
vo ng ch U th nht, v chun bng natri hydroxyd
0,1 N ti khi c mu vng sng, lc nh. Sau khi chun
ng ch U th nht, tho ng ni, lp n vi ng ch U th
hai u ra, v chun tng t. Ghi tng lng dung
dch hai ln chun l Vt (ml).
Tin hnh lm mt mu trng, th tch dung dch natri
hydroxyd 0,1 N l Vo. Tnh phn trm lu hunh dioxyd
trong mu th theo cng thc:
SO 2 %
(Vt
Vo ) 0,0032
100
m
Trong
m l khi lng ca mu th (g)
Nit amoniac
Cho 25 ml acid sulfuric 0,1 N vo bnh hng 500 ml, v lp

vi thit b ct gm ng ni bu Kjeldahl v mt sinh hn
c u ra ngp pha di b mt dung dch trong bnh
hng. Cho khong 2 g cht mu caramel, cn chnh xc,
vo trong bnh Kjeldahl c di 800 ml, v thm vo bnh 2
g magnesi oxyd (khng c carbonat), 200 ml nc, v vi
vin bt. Lc bnh trn u hn hp trong bnh, v
lp nhanh bnh vi thit b ct. un nng bnh ti si, thu
ly khong 100 ml dch ct vo bnh hng. Ra u ra ca
ng sinh hn vi vi ml nc, cho dch ra vo bnh hng,
sau thm 4 hoc 5 git ch th methyl (500 mg
methyl trong 100 ml alcol), v chun bng natri hydroxyd
0,1 N, ghi th tch l Vt (ml). Tin hnh lm mt mu trng,
th tch dung dch natri hydroxyd 0,1 N trung tnh l Vo.
Tnh phn trm nit amoniac trong mu th theo cng
thc:
Nit amoniac
(Vt
Vo ) 0,0014
100
m
Trong
m l khi lng ca mu th (g)
4-Methylimidazol
Ghi ch: Cn thng tin v mt phng php tt hn.

Dng cc nguyn vt liu v thuc th sau (thuc th phi
l loi ACS hoc tng ng).
Nguyn vt liu:
Bng thy tinh loi Pyrex, ct sc k 22 300 mm v kho
PTFE (nh Kimax 17800); cc polypropylen 150 ml (nh
Nalge 1201); bnh y trn 250 ml (nh Pyrex 4320); phu
rt bt 75 mm; tha gt (spatula) 5 cm; my ct quay chn
khng, bp a kn, ni cch thy, pipet Pasteur dng mt
ln; bnh nh mc 5 ml.
Thuc th:
Aceton; Celit 545; methylen clorid; natri hydroxyd; v
tetrahydrofuran.
Tin hnh:
Sau khi trn k mu th cht mu caramel bng cch lc
hoc khuy, cn 10,00 g hn hp vo cc polypropylen 150
ml. Polypropylen c coi l tt hn thy tinh v b mt k
nc d chuyn ton lng mu th. Thm vo 5,0 g
dung dch NaOH 3,0 N v lc k m bo pH ca mu
th ln hn 12. Thm 20 g Celit 545 vo cc trn, v trn
u ti khi hn hp gn kh li. Qu trnh ny khong 2 ti
3 pht. Vi mu th c hm lng nc qu cao, hn hp
cht mu caramel-Celit 545 c th qu t. Trong trng
hp ny, trn 5,00 g hn hp cht mu caramel vi 2,5 g
NaOH 3,0 N v 15 g Celit 545 v tin hnh phn tch.
Mt nt bng thy tinh Pyrex c t pha di ct sc
k 22
300 mm vi kha PTFE. Hn hp cht mu
caramel-Celit 545 c cho vo ct qua phu 75 mm. Hn
hp trong ct c nhi bng cch g nh ct theo
phng thng ng khong 10 cm trn mt b mt c lt.
Khi nhi tt, hn hp cht mu caramel-Celit 545 phi
chim xp x 250 mm pha di ct. Cn thn khi tin hnh
trnh nhi lng qu hoc cht qu. Ct nhi lng qu
methylen clorid s ra gii qu nhanh v khng chit hon
ton. Ct nhi qu cht dn ti dung mi ra gii kh tip
cn mt s vng cht nhi. iu ny cng dn n vic
chit khng hon ton.
M kha, dng cc rt methylen clorid vo ct. Khi dung
mi ti nt bng thy tinh, ng kho v dung mi
tng tc vi hn hp trong ct 5 pht. Sau m kha v
ct c ra gii tip vi methylen clorid ti khi thu
c 200 ml vo bnh y trn 250 ml. Ly 1,00 ml dung
dch chun ni 2 MEI (50,0 mg 2 MEI/50,0 ml methylen
clorid) thm vo dch ra gii thu c. 2 MEI c
tch tt khi 4 MEI trong iu kin sc k kh lng dng
v khng thy c trong cht mu caramel.
Sau , loi dung mi khi dch ra gii bng ct quay p
sut gim 45-50 kPa v bnh cu y trn c duy tr
35 oC trong ni cch thy. Cn ca dch chit c chuyn
hon ton sang bnh nh mc 5 ml bng pipet Pasteur
dng mt ln, bng cch ra bnh cu nhiu ln mi ln vi
mt lng nh (khong 0,75 ml) tetrahydrofuran hoc
aceton. C hai dung mi c dng vi kt qu nh nhau.
Sau khi trn k hn hp bng cch lc ngc bnh nhiu
ln, c dch chit dng cho phn tch GLC. Dch chit
sau khi chun b phi phn tch cng sm cng tt, bi
mu ch n nh trong vng 1 ngy sau khi chun b.
Phn tch GLC c tin hnh bng thit b sc k kh vi
detector ngn la hydro. Ct thy tinh 1 m 6 mm ng
knh ngoi ( 4 mm ng knh trong), thm 7,5% Carbowax
20M + 2% KOH trn 90/100 mesh Anakrom ABS. Cc
thng s GLC nh sau: kh mang nit 50 ml/pht; hydro 50
ml/pht; oxy 80 ml/pht; b phn tim mu 200 oC; ct
ng nhit 180 oC; detector 250 oC; th tch mu 5 L. Tt
c qu trnh nh lng tin hnh dng k thut chun ni.
2-Acetyl-4-tetrahydroxy- Ghi ch: Cn thng tin v mt phng php tt hn.
butylimidazol (THI)
THI
c
chuyn
thnh
dn
xut
2,4dinitrophenylhydrazon (THI-DNPH) ca n. Dn cht ny
c tch khi thuc th d v tp carbonyl bng HPLC
dng ct RP-8, xc nh bng o hp th bc sng
385 nm.
Tin hnh:
Cn chnh xc cht mu caramel (200-250 mg), sau ha
tan trong nc (3 ml). Chuyn ton lng dung dch sang
phn pha trn ca ct t hp. Bt u ra gii bng nc,
v ton b lng nc qua cc ct khong 100 ml.
Sau tho ct pha trn. Ra gii ct pha di bng HCl
0,5 N. B 10,0 ml dch ra gii ban u, sau thu ly 35

ml.
C dung dch ny ti kh 40 oC v 15 torr. Ha tan cn
c st trong methanol khng c carbonyl (250 L) v
thuc th 2,4-dinitrophenylhydrazin (250 L). Chuyn hn
hp phn ng sang l c np l tm m v bo qun 5
gi nhit phng. Tim 5 L (cng c th t 1 ti 25
L) vo ct HPLC LiChrosorb RP-8 (10 m). Pha ng gm
MeOH/ H3PO4 0,1 M 50/50 (tt/tt). iu chnh thnh phn
pha ng nu c tnh ca ct thay i, tu thuc nh sn
xut. (Rt nn s dng cht nhi LiChrosorb RP-8, 10 m,
ct 250 ( 4 mm "Vertex" ca nh sn xut Knauer, Bad
Homburg, F.R.G.). Vi tc pha ng 2 ml/pht v kch
c ct 250 ( 4,6 mm, THI-DNPH ra gii sau khong
6,30,1 pht. Pht hin 385 nm v o chiu cao ca pic.
Hm lng c tnh da vo ng chun ca THIDNPH trong methanol.
Nguyn vt liu:
Thuc th - 2,4,-Dinitrophenylhydrazin hydroclorid: 2,4dinitrophenylhydrazin thng mi (5 g) cho vo acid
hydrocloric c (10 ml) trong bnh nn 100 ml, v lc nh
ti khi base t do (mu ) chuyn thnh hydroclorid (mu
vng). Thm vo ethanol (100 ml), lc v un nng trn
cch thy si ti khi tt c cht rn ha tan. Sau khi kt tinh
nhit phng, hydroclorid c lc, ra bng ether,
sy nhit phng v bo qun trong bnh ht m. Trong
khi bo qun, hydroclorid dn chuyn thnh base t do.
Sau , c th loi bng cch ra vi dimethoxyethan.
Thuc th c chun b bng cch trn 0,5 g 2,4dinitrophenylhydrazin hydroclorid trong 15 ml methanol 5%
trong dimethoxyethan trong 30 pht. Bo qun trong t lnh
v kim tra nh k bng HPLC.
- Nha trao i cation (mnh): Dowex 50 AG x 8, H+, 100200 mesh.
- Nha trao i cation (yu): Amberlite CG AG 50 I, H+,
(100-200 mesh). ( lng hai hoc ba ln trc khi s
dng).
- Methanol, khng c carbonyl t do: Methanol c chun
b theo Y. Peleg v C.H. Mannheim, J. Agr. Fd. Chem, 18
(1970) 176, bng cch x l vi thuc th Girard P.
- Dimethoxyethan: Nu khng tinh khit, dimethoxyethan
c tinh ch bng cch ct t 2,4-dinitrophenylhydrazin
vi s c mt ca acid v ct li t natri hydroxyd. Ngay
trc khi s dng cho qua ct nhm trung tnh loi
peroxyd.
Thit b:
Ct t hp: tng t nh m t trong J. Agr. Fd. Chem.
22 (1974) 110. Ct pha trn (150 ( 12,5 mm, chiu cao
nhi ti a 9 cm, hoc 200 ( 10 mm, chiu cao nhi ti a
14 cm, vi mao qun u ra c 1 mm ng knh trong)
nhi cht trao i cation acid yu, chiu cao cht nhi ln
lt khong 50-60, hoc 80-90 mm. Ct pha di (chiu
di tng 175 mm, ng knh trong 10 mm, vi u ra mao
qun v kho Teflon) nhi vi cht trao i cation acid
mnh chiu cao cht nhi 60 mm. Bnh ng dung mi,
dng phu nh git (100 ml) kha Teflon. Tt c cc phn
c ni bng khp ni thy tinh mi chun (14,5 mm).
HPLC: vi ct qui nh trn v detector UV c kh nng
o 385 nm.
Chun ha: Ha tan THI-DNPH trong methanol tuyt i
khng c carbonyl (khong 100 mg/L; nng ca THI:
47,58 ng/L). Dung dch ny pha long 10 ln vi methanol
(4,7 ng THI/L). Dung dch chun THI-DNPH n nh t nht
20 tun nu bo qun trong t lnh.
Arsen
Ch

- Phng php II.
- S dng k thut hp th nguyn t thch hp cho hm
lng quy nh xc nh. La chn c mu th v
Ph lc 20
YU CU K THUT V PHNG PHP TH I VI BRILLIANT BLACK PN
1. Tn khc, ch s
CI food Black 1, Black PN; CI (1975): 28440

INS: 151
2. nh ngha
Ch yu gm tetranatri 4-acetamido-5-hydroxy-6-[7-sulfon4-(4-sulfon-phenylazo)-1-naphthylazo]-1,7-naphthalendisulfonat cng cc cht mu ph cng vi NaCl v (hoc)

Na2SO4 l cc thnh phn khng mu chnh.
Tn ha hc
tetranatri 4-acetamido-5-hydroxy-6-[7-sulfon-4-(4-sulfonphenylazo)-1-naphthylazo]-1,7-naphthalen-disulfonat.
M s C.A.S.
2519-30-4
Cng thc ha hc
C28H17N5Na4O14S4
Cng thc cu to
NHCOCH3
HO
NaO3S
N
SO3Na
NaO3S
SO 3Na
Khi lng phn t
867,69
3. Cm quan
Dng bt hoc tinh th mu en
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 4,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,8% tng cc acid 4-Acetamido-5hydroxy-1,7-naphthalendisulfonic; acid 4-Amino-5-hydroxy1,7-naphthalendisulfonic; acid 8-Amino-2naphthalensulfonic; acid Sulfanilic v acid 4,4'-Diazoaminodi(benzensulfuric)

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Ch

Cc cht mu ph

- S dng cc iu kin sau:
Dung mi khai trin: Sc k (i): s 1
Sc k (ii): s 4
Chiu cao tuyn dung mi: Sc k (i): 17 cm
Sc k (ii): 17 cm.)
Cc cht hu c ngoi
cht mu

- S dng k thut sc k lng, vi cc iu kin nh sau:
HPLC ra gii gradient t 2 n 100%, tc tng c nh
2%/pht (tuyn tnh).
6.2. nh lng
Cn 0,6 - 0,7 g mu th;
mu (D).
Ph lc 21
YU CU K THUT V PHNG PHP TH I VI BROWN HT
1. Tn khc, ch s
CI food Brown 3, Choccolat brown HT; CI (1975): 20285

INS: 155
2. nh ngha
Ch yu gm dinatri 4,4'- (2,4-dihydroxy-5-hydroxymethyl1,3-phenylen-bisazo) di-1-naphthalen-sulfonat cng cc

cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh
phn khng mu chnh.
Tn ha hc
Dinatri 4,4'- (2,4-dihydroxy-5-hydroxymethyl-1,3-phenylenbisazo) di-1-naphthalen-sulfonat.
M s C.A.S.
4553-89-3
Cng thc ha hc
C27H18N4Na2O9S2
Cng thc cu to
NaO 3S
HO
N N
HO
CH2OH
SO 3Na
Khi lng phn t
652,57
3. Cm quan
Dng bt hoc tinh th mu nu.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, khng tan trong ethanol.
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135 oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%.
Ch
Cc cht mu ph
Khng c qu 10,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,7% acid 4-Aminonaphthalen-1-sulfonic.

khng sulfon ho
Cc cht c th chit
bng ether
Khng c qu 0,2%.
5.3. Hm lng
Khng c thp hn 70,0% tng cc cht mu
6. Phng php th
6.1 tinh khit
Ch

Cc cht mu ph

- S dng cc iu kin sau:
Pha chun theo cch sau: Pha long 1,0 ml dung dch ch
phm 1% vi nc n 100 ml, lc u. Ly 0,10 ml
dung dch ny vo ng nghim, thm 5,0 ml dung dch
nc:aceton (1:1 - tt/tt) v sau thm 14,9 ml dung dch
natri hydrocarbonat 0,05 N, lc u. Xc nh hp th
ca chun - As.
Dung mi khai trin: S 6.
Thi gian khai trin: ~ 14 gi.
Cc cht hu c ngoi
cht mu

- S dng k thut sc k lng, vi cc iu kin nh sau:
HPLC ra gii gradient t 2 n 100%, tc tng c nh
2%/pht.
6.2. nh lng
hm lng bng quang ph (trong JECFA monograph 1Vol. 4) vi cc iu kin nh sau:
m: m phosphat pH 7;
Pha long dung dch A: 10 ml pha long n 250 ml.
hp th ring (a): 40,3
Bc sng cc t hp th: ~ 460 nm.
Ph lc 22
YU CU K THUT V PHNG PHP TH I VI
-CAROTEN TNG HP
1. Tn khc, ch s
CI food Orange 5; CI (1975): 40800

INS: 160ai
2. nh ngha
Ch phm ch yu gm tt c cc ng phn trans (Z) ca

-caroten vi lng nh cc carotenoid khc; Cc dng
pha long v n nh c chun b t -caroten p ng
quy chun ny gm dung dch hoc huyn ph ca caroten trong du, m thc phm, nh tng v bt c th
phn tn trong nc; Cc ch phm ny c th c t s
gia cc ng phn cis/trans khc nhau; Phng php
phn tch i vi cht mu gc khng thch hp nh
lng hoc xc nh tinh khit ca cc ch phm n
nh (cc phng php ny c th c cc nh sn xut
cung cp).
Tn ha hc
-caroten; , -caroten
M s C.A.S.
7235-40-7
Cng thc ha hc
C40H56
Cng thc cu to
H3C
CH3
CH3
CH3
H3C
CH3
CH3
CH3
H3C
CH3
Khi lng phn t
536,88
3. Cm quan
Tinh th hoc bt tinh th mu n nu, nhy cm

vi oxy v nh sng do vy phi bo qun trong bao b
trnh nh sng v mi trng kh tr.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc, thc t khng tan trong ethanol, t

tan trong du thc vt, tan trong cloroform.
Quang ph
Xc nh hp th quang (A) ca dung dch mu th C

(xem phn nh lng) ti cc bc sng 455 nm, 483 nm
Xc nh hp th quang (A) ca dung dch mu th C ti
bc sng 455 nm v ca dung dch mu th B ti bc
sng 340 nm
T s A455/A340 khng c thp hn 15.
Carotenoid
Phi c phn ng c trng ca carotenoid.
Phn ng Carr-Price
Dung dch mu th trong cloroform chuyn thnh mu xanh

lam khi thm d dung dch thuc th Carr-Price (TT).
5.2. tinh khit

Tro sulfat
Khng c qu 0,1%.
Cc cht mu ph
Cc carotenoid khc ngoi

3,0% tng cc cht mu.
Ch
-caroten: khng c qu
5.3. Hm lng -caroten Khng c thp hn 96% tng cht mu.

6. Phng php th
6.1. nh tnh
Carotenoid
Mu ca dung dch mu th trong aceton s mt sau khi

thm dung dch natri nitrit 5% v dung dch acid sulfuric 1
N, lc u.
6.2. tinh khit

Tro sulfat

- Phng php I, cn 2 g mu th
Cc cht mu ph
Cc carotenoid khc ngoi -caroten

Cn 80 mg mu, ha tan trong 100 ml cloroform. Chm 400
L dung dch ny ln bn mng (silicagel 0,25 mm), chm
cch y bn mng 2 cm. Ngay sau khi chm tin hnh
khai trin sc k trong bung bo ha h dung mi gm
95 phn dicloromethan v 5 phn diethyl ether, trnh nh
sng, n khi chiu cao tuyn dung mi cch vt chm
khong 15 cm. Ly bn mng ra, kh dung mi ti nhit
phng, nh du vt mu chnh v cc vt mu ph
(cc carotenoid khc).
Co phn silicagel hp ph cht mu chnh, cho vo mt
ng ly tm 100 ml c nt thy tinh, thm 40,0 ml cloroform
(dung dch 1).
Co phn silicagel hp ph cc carotenoid khc, cho vo
mt ng ly tm 50 ml c nt thy tinh, thm 20,0 ml
cloroform (dung dch 2).
Lc c hai ng trong 10 pht v ly tm trong 5 pht. Ly
10,0 ml dung dch 1 pha long vi cloroform n 50,0 ml
(dung dch 3). S dng thit b o quang thch hp, xc
nh hp th quang ca dung dch 2 v dung dch 3 ti
bc sng 464 nm, s dng cloroform lm mu trng v o
trong cuvet 1 cm.
Tnh kt qu:
Tnh hm lng (%) tng cc carotenoid khc ngoi
caroten theo cng thc sau:
Ch
Trong :
A2 = hp th quang ca dung dch 2
6.2. nh lng
Tin hnh theo hng dn trong chuyn lun nh lng
tng cht mu bng phng php quang ph trong JECFA
monograph 1-Vol. 4, vi cc iu kin nh sau:
W = 0,08 g
V1 = V2 = V3 = 100 ml
v1 = v2 = 5 ml
A1%1cm = 2500
max
~ 456 nm.
Ph lc 23
YU CU K THUT V PHNG PHP TH
I VI CAROTEN THC VT
1. Tn khc, ch s
-Caroten t nhin; caroten t nhin; hn hp cc caroten;

CI food Orange 5; S CI (1975): 75130, S CI (1975):
40800 (i vi -caroten).
INS 160aii
ADI chp nhn c c ngha l mc s dng khng
vt qu mc thng thng c trong thc vt.
2. nh ngha
Cc caroten (thc vt) thu c bng cch chit c rt

(Daucus carota), du c (Elaeis guinensis), khoai lang
(Ipomoea batatas) v cc thc vt n c khc vi dung
mi hu c, sau tinh ch. Cht mu chnh l - v caroten, trong ch yu l -caroten. Ngoi ra cn c
lng nh - v -caroten cng cc thnh phn mu khc.
Bn cnh cc thnh phn mu, ch phm cn c th cha
du, m, sp t nhin c ngun gc t nguyn liu. Ch
nhng dung mi sau dy c s dng chit caroten:
aceton, methanol, ethanol, propan-2-ol, hexan, carbon
dioxyd v du thc vt.
Ch phm thng mi ch yu tn ti di dng dung dch
hoc huyn ph trong du thc vt thc phm. Dng ny
d s dng v gip ci thin tnh bn ca caroten d b oxy
ha.
Carotenoid
M s C.A.S.
7235-40-7
Cng thc ha hc
C40H56 (- caroten)
Cng thc cu to
H3C
CH3
CH3
CH3
H3C
CH3
CH3
CH3
H3C
CH3
All-trans- -caroten
Khi lng phn t
536,88 ( -caroten)
3. Cm quan
Dng rn hoc dng lng mu nu n nu hoc cam

n cam sm.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc.
Quang ph
Dung dch mu th trong n-hexan (1/200.000) c cc i

hp th ti vng 440 - 457 nm v 470 - 486 nm.
Phn ng mu
5.2. tinh khit

Dung mi tn d
Ch
5.3. Hm lng
Khng c qu 50,0 mg/kg n cht hoc tng ca

aceton, hexan, methanol, ethanol, propan-2-ol.
Hm lng caroten (tnh theo -caroten) khng c thp
hn hm lng c cng b.
6. Phng php th
6.1 nh tnh
Phn ng mu
Chm ln tm giy lc dung dch mu th trong toluen (~

400 g -caroten/ml), phun hoc nh dung dch antimon
triclorid 20% trong toluen ln vt mu trn giy lc, sau 2 3 pht mu ca vt chuyn thnh xanh lam.
6.2 tinh khit

Ch

m t trong JECFA monograph 1-Vol. 4 phn cc phng
php phn tch cng c.
6.3. nh lng
W = Lng mu th o hp th quang (g)
V1 = V2 = V3 = 100 ml
v1 = v2 = 5 ml
A1%1cm = 2500
max = 440 - 457 nm.
Ph lc 24
YU CU K THUT V PHNG PHP TH I VI CAO ANNATTO
1. Tn khc, ch s
Annatto F; Orlean; Terre orellana; R. Orange; S CI (1975):

75120 (Natural Orange 4).
INS 160b
ADI = 0 - 12 mg/kg th trng i vi Bixin;
ADI = 0 - 0,6 mg/kg th trng i vi Norbixin v cc mui
ca n
Mu Bixin v Norbixin c ch t cao Annatto (bixin c
chit bng dung mi, norbixin c chit bng dung mi;
bixin x l vi nc; norbixin x l vi kim, kt ta acid;
v norbixin x l vi kim, khng kt ta acid) u p dng
ADI i vi Bixin v Norbixin.
2. nh ngha
Norbixin x l vi kim, kt ta acid c sn xut bng

cch loi b v ca ht cy Annatto (Bixa orellana L.) bng
dung dch kim. Bixin b thy phn thnh norbixin trong
dung dch kim nng, sau acid ha kt ta norbixin.
Lc ta, lm kh v nghin thu c dng bt, ht.
Norbixin x l kim cha nhiu thnh phn mu, cc thnh
phn mu chnh (chim t l ln) l cis-norbixin, ngoi ra
cn c lng nh trans-norbixin v cc sn phm phn hy
nhit ca norbixin l sn phm ph ca qu trnh ch bin.
Ch phm thng mi dng cho cng nghip thc phm
thng c bo ch sn vi cc cht mang thch hp vi
cht lng dng cho thc phm.
Tn ha hc
cis-Norbixin: acid 6,6'-Diapo-,-carotendioic

Mui dikali cis-Norbixin: Dikali 6,6'-diapo-,-carotendioat
Mui dinatri cis-Norbixin: Dinatri 6,6'-diapo-,carotendioat
cis-Norbixin: 542-40-5
Mui dikali cis-Norbixin: 33261-80-2
Mui dinatri cis-Norbixin: 33261-81-3
C24H28O4; C24H26K2O4; C24H26Na2O4
M s C.A.S.
Cng thc ha hc
Cng thc cu to
Khi lng phn t
380,5 (acid); 456,7 (mui dikali); 424,5 (mui dinatri)
3. Cm quan
Bt mu nu sm n tm.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong dung dch kim, t tan trong ethanol.
Hp th UV-VIS
Dung dch mu th 0,5% trong dung dch kali hydroxyd c

cc i hp th ti khong 453 v 482 nm.
Sc k lp mng
5.2. tinh khit

Arsen
Ch
Thy ngn
5.3. Hm lng
Tng cc cht mu khng thp hn 35,0% (tnh theo

norbixin)
6. Phng php th
6.1 nh tnh
Sc k lp mng
Hot ha bn mng (VD: LK6D SILICA GEL 60 A (chiu

dy lp pha tnh: 250 m, c bn mng: 5 x 20 cm)) trong 1
gi ti 110o. Pha dung dch mu th 5% trong ethanol
(95%) v chm 10 L ln bn mng. kh v khai trin
sc k trong h dung mi gm n-butanol/ methyl ethyl
keton/dung dch amoniac 10% (3/2/2 - tt/tt/tt) n khi chiu
cao ca tuyn dung mi so vi vt chm khong 10 cm.
Ly bn mng ra v kh. Vt bixin v norbixin c mu
vng vi gi tr Rf tng ng khong 0,50 n 0,45. Phun
dung dch natri nitrit 5%, tip theo phun acid sulfuric 0,5 M
vt s mt mu ngay.
6.2 tinh khit

Arsen

- S dng k thut ICP-AES / AAS-hydrid. Hoc xc nh
theo chuyn lun th gii hn Arsen - phng php II. La
nguyn tc ca phng php m t trong JECFA
monograph 1-Vol. 4.
Ch

- S dng k thut AAS ICP-AES thch hp vi mc hm
lng nh xc nh. La chn c mu th v
phng php m t trong JECFA monograph 1-Vol. 4.
Thy ngn

- S dng k thut AAS ha hi lnh. La chn c mu
th ph hp vi mc hm lng quy nh.
6.3. nh lng
monograph 1-Vol. 4 - quy trnh 1, vi cc iu kin nh
sau:
Dung mi: dung dch KOH 0,5%
o hp th quang ti max ~ 482 nm.
hp th ring A1%1cm = 2870.
Ph lc 25
YU CU K THUT V PHNG PHP TH I VI -apo-8-CAROTENAL
1. Tn khc, ch s
CI food Orange 6; S CI (1975): 40820

INS 160e
2. nh ngha
L ch phm ch yu gm ng phn all trans (Z) ca apo-8-carotenal vi lng nh cc carotenoid khc; Cc

dng pha long v n nh c chun b t -apo-8carotenal p dng quy chun ny gm dung dch hoc
huyn ph ca -apo-8-carotenal trong du, m thc
phm, nh tng v bt phn tn trong nc; Cc ch
phm ny c th c t s gia cc ng phn cis/trans
khc nhau; Phng php phn tch i vi cht mu gc
khng thch hp nh lng hoc xc nh tinh khit
ca cc dng n nh ch phm (cc phng php thch
hp s c cc nh sn xut cung cp).
Tn ha hc
-apo-8-Carotenal, 8-apo- -carotenal
M s C.A.S.
1107-26-2
Cng thc ha hc
C30H40O
Cng thc cu to
H3C
CH3
CH3
CH3
CH3
CHO
CH3
CH3
Khi lng phn t
416,65
3. Cm quan
Tinh th hoc bt tinh th mu tm m c nh kim, nhy

cm vi oxy v nh sng do vy phi bo qun trong bao
b trnh nh sng v trong mi trng kh tr.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc, kh tan trong ethanol, t tan trong

du thc vt, tan trong cloroform.
Quang ph
Xc nh hp th quang (A) ca dung dch mu th

(xem phn nh lng) ti cc bc sng 461 nm v 488
nm
Carotenoid
Phi c phn ng nh tnh ca carotenoid.
Phn ng Carr-Price

5.2. tinh khit

Tro sulfat
Khng c qu 0,1%.
Cc cht mu ph
Khng c qu 3,0% tng cc cht mu.

Xem hng dn trong phn phng php th.
Ch
5.3. Hm lng
6. Phng php th
6.1. nh tnh
Carotenoid

thm dung dch natri nitrit 5% v dung dch acid sulfuric 1
N, lc u.
6.2. tinh khit

Tro sulfat
- Th theo hng dn ti JECFA monograph 1 - Vol.4

- Phng php I, cn 2 g mu th
Cc cht mu ph
Cn 80 mg mu, ha tan trong 100 ml cloroform. Chm 400

L dung dch ny ln bn mng cch y 2 cm (silicagel
0,25 mm), bn mng c x l trc nh sau: nhng
bn mng vo b cha dung dch KOH 3% trong methanol
sao cho thm t u, kh ngoi khng kh trong 5 pht
v hot ha bng cch sy trong t sy ti 110 oC trong 1
gi, ngui v gi trong bnh ht m cha CaCl2.
Ngay sau khi chm tin hnh khai trin sc k trong bung
bo ha h dung mi gm n-hexan/cloroform/ethyl
acetat (70/20/10), trnh nh sng, n khi chiu cao tuyn
dung mi cch vt chm khong 10 cm. Ly bn mng ra,
kh dung mi ti nhit phng, nh du vt mu
chnh v cc vt mu ph (cc carotenoid khc).
(dung dch 1).
(dung dch 3). S dng thit b o quang ph thch hp, xc
trong cuvet 1 cm.
Tnh kt qu:
Tnh hm lng (%) tng cc carotenoid khc ngoi -apo8carotenal theo cng thc sau:
Ch
Trong :
6.3. nh lng
Cn khon 80 mg (chnh xc n mg) mu th. Tin hnh
theo hng dn trong chuyn lun nh lng tng cht
mu bng phng php quang ph trong JECFA
hp th ring (a) = 2640
max
~ 461 nm.
Ph lc 26
YU CU K THUT V PHNG PHP TH I VI ESTER METHYL
(HOC ETHYL) CA ACID -APO-8-CAROTENOIC
1. Tn khc, ch s
CI food Orange 7; S CI (1975): 40825

INS 160f
2. nh ngha
L ch phm ch yu gm ng phn all trans (Z) ca Ethyl

ester ca acid -apo-8-carotenoic vi lng nh cc
carotenoid khc; Cc dng pha long v n nh c
chun b t Ethyl ester ca acid -apo-8-carotenoic p
ng quy chun ny gm dung dch hoc huyn ph ca
Ethyl ester ca acid -apo-8-carotenoic trong du, m thc
phm, nh tng v bt c th phn tn trong nc; Cc
ch phm ny c th c t s gia cc ng phn cis/trans
khc nhau; Phng php phn tch i vi cht mu gc
khng thch hp nh lng hoc xc nh tinh khit
ca cc dng n nh ca ch phm (cc phng php ny
s c cc nh sn xut cung cp).
Tn ha hc
Ethyl ester ca acid -apo-8-carotenoic; Ethyl 8-apo- caroten-8-oat
M s C.A.S.
1109-11-1
Cng thc ha hc
C32H44O2
Cng thc cu to
H3C
CH3
CH3
CH3
O
OC2H5
CH3
CH3
CH3
Khi lng phn t
460,70
3. Cm quan
Tinh th hoc bt tinh th mu n tm, nhy cm

vi oxy v nh sng do vy phi bo qun trong bao b
trnh nh sng v mi trng kh tr.
4. Chc nng
Phm mu.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc, rt kh tan trong ethanol, kh tan

trong du thc vt, tan trong cloroform.
Quang ph
Xc nh hp th quang (A) ca dung dch mu th

(xem phn nh lng) ti cc bc sng 449 nm, 475 nm
Carotenoid
Phi c phn ng c trng ca carotenoid.
Phn ng Carr-Price

5.2. tinh khit

Tro sulfat
Khng c qu 0,1%.
Cc cht mu ph
Khng c qu 3,0% tng cc cht mu.
Ch
5.3. Hm lng
6. Phng php th
6.1. nh tnh
Carotenoid

thm lin tip dung dch natri nitrit 5% v dung dch acid
sulfuric 1 N, lc u.
6.2. tinh khit

Tro sulfat
Th theo hng dn ti JECFA monograph 1-Vol. 4 Phng php I, cn 2 g mu th
Cc cht mu ph
Cc carotenoid khc ngoi Ethyl ester ca acid -apo-8carotenoic: Cn 80 mg mu, ha tan trong 100 ml
cloroform. Chm 400 L dung dch ny ln bn mng cch
y 2 cm (silicagel 0,25 mm), bn mng c s l
trc nh sau: nhng bn mng vo b cha dung dch
KOH 3% trong methanol sao cho thm t u, kh
ngoi khng kh trong 5 pht v hot ha bng cch sy
trong t sy ti 110 oC trong 1 gi, ngui v gi trong
bnh ht m cha CaCl2.
Ngay sau khi chm tin hnh khai trin sc k trong bung
bo ha h dung mi gm n-hexan/cloroform/ethyl
acetat (70/20/10), trnh nh sng, n khi chiu cao tuyn
dung mi cch vt chm khong 10 cm. Ly bn mng ra,
kh dung mi ti nhit phng, nh du vt mu
chnh v cc vt mu ph (cc carotenoid khc).
(dung dch 1).
(dung dch 3). S dng thit b o quang ph thch hp, xc
trong cuvet 1 cm.
Tnh kt qu:
Tnh hm lng (%) tng cc carotenoid khc ngoi Ethyl
ester ca acid -apo-8-carotenoic theo cng thc sau:
Ch
Trong :
6.3. nh lng
Cn khon 80 mg (chnh xc n mg) mu th. Tin hnh
theo hng dn trong chuyn lun nh lng tng cht
mu bng phng php quang ph trong JECFA
hp th ring (a) = 2550
max
~ 449 nm.
Ph lc 27
YU CU K THUT V PHNG PHP TH I VI CAO V NHO
1. Tn khc, ch s
Enociania, Eno, Grape skin extract.

INS 163 (ii)
ADI = 0-2,5 mg /kg th trng.
2. nh ngha
Ch phm thu c t dch chit nc ca v hoc b nho

sau khi p ht nc qu; cha cc thnh phn thng
thng ca nc p qu nho nh: anthocyanin, acid
tartric, tanin, cc ng, khong cht,, nhng t l thnh
phn khc so vi trong nc p qu. Trong qu trnh chit,
lu hunh dioxyd c cho vo v hu ht lng ng
chit c b ln men bin thnh ru (alcol); Dch chit
c c c bng cch lm bay hi trong chn khng, trn
thc t trong qu trnh ny ton b lng alcol b loi i;
mt lng nh lu hunh dioxyd c th cn d li.
Tn ha hc
Cc cht mu chnh l anthocyanin, glucosid ca cc

anthocyanidin (cc mui 2-phenylbenzopyrylium) nh
peonidin, malvidin, delphinidin v petunidin.
Cng thc ha hc
Peonidin : C16H13O6 X
Malvidin : C17H15O7 X
Delphinidin : C15H11O7 X
Petunidin : C16H13O7 X
X : Gc acid
Cng thc phn t
R
OH
+
HO
R'
OH
OH
Peonidin : R = OCH3 ; R= H
Malvidin : R, R= OCH3
Delphinidin : R, R= OH
Petunidin : R = OCH3 ; R= OH
X : Gc acid
3. Cm quan
Cht lng, ming, bt hoc bt nho mu ta, c mi c

trng nh.
4. Chc nng
Phm mu
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc
Quang ph
Dch chit pH 3 cho cc i hp th ti ~ 525 nm
Phn ng mu
5.2. tinh khit

Lu hunh dioxyd
Khng c qu 0,005% vi 1 ch s mu.
Cc cht mu kim
Cc cht mu acid khc t yu cu (m t trong phn Phng php th)

Ch
Khng c qu 2,0 mg/kg
5.3. Hm lng
Cng mu khng c di mc cng b.
6. Phng php th
6.1 nh tnh
Phn ng mu
Thm 0,1 g mu vo 50 ml nc v lc k. Lc nu cn,

dung dch c mu n ta v chuyn sang mu da
tri hoc mu xanh l sm khi thm dung dch natri
hydroxyd (TT).
6.2. tinh khit

Lu hunh dioxyd
Ct 1 g mu vi 100 ml nc v 25 ml dung dch acid

phosphoric (2 /7) trong mt bnh ct vi ng Wagner (Hnh
1). Cho 25 ml dung dch ch acetat (1/50) pha trc vo
mt bnh hp th. Nhng phn thp nht ca ng ngng
ngp vo dung dch ch acetat trong bnh hp th. Ct n
khi cht lng trong bnh hp th c khong 100 ml v
ra phn ui ng ct bng mt lng nh nc. Thm 5
ml dung dch acid hydrocloric v 1 ml dung dch h tinh bt
(TT) vo dch ct ri chun bng dung dch iod 0,01 N.
Mi ml dung dch iod 0,01 N tng ng vi 0,3203 mg SO2
Hnh 1 : ng Wagner
Cc cht mu kim
Cho 1 g mu vo 100 ml dung dch natri hydroxyd (1/100)

v lc u. Ly 30 ml dung dch ny v chit vi 15 ml
ether. Lc dch chit ether ny hai ln mi ln vi 5 ml dung
dch acid acetic long (TT). Dch chit acid acetic khng
c c mu.
Cc cht mu acid khc Thm 1 ml dung dch amoniac (TT) v 10 ml nc vo 1 g

mu v theo ch dn trong phn sc k, chm 0,002 ml
dung dch ln bn sc k v lm kh. Dng dung mi khai
trin l hn hp pyridin v amoniac (TT) (2:1, theo th
tch), dung mi khai trin c 15 cm t vt chm.
Sau khi lm kh bn mng di nh sng thng, khng
c xut hin vt tuyn dung mi. Nu c bt c vt
no th vt phi mt mu ngay khi phun bn mng vi
dung dch thic (II) clorid (SnCl2) trong acid hydrocloric
(2/5).
Ch
Th theo hng dn ti JECFA monograph 1-Vol. 4). S

dng k thut hp th nguyn t thch hp cho hm lng
quy nh xc nh. La chn c mu th v phng
m t trong JECFA monograph 1-Vol. 4: cc phgn php
th chung phn cc phng php phn tch dng c.
6.3. nh lng
Trong iu kin khng c phng php nh lng, c th
dng phng php o cng mu nh sau:
Chun b khong 200 ml dung dch m pH 3 acid citricNa2HPO4 : trn 159 th tch dung dch acid citric 2,1% v
41 th tch dung dch Na2HPO4 0,16% ri chnh pH v 3
bng dung dch acid citric hoc dung dch Na2HPO4.
Cn chnh xc mt lng thch hp mu th sao cho mt
quang o c trong khong t 0,2 n 0,7 v thm
dung dch m pH 3 acid citric-dinatri hydrophosphat n
va 100 ml. o hp th A ca dung dch trong mt
cuvet o 1 cm, bc sng cc i hp th 525 nm, dng
dung dch m pH 3 acid citric-dinatri hydrophosphat lm
mu trng.
Ch s mu =
A 10
m
m = Khi lng mu th (g)
Ph lc 28
YU CU K THUT V PHNG PHP TH I VI TITAN DIOXYD
1. Tn khc, ch s
Titania, CI pigment white 6, CI (1975) No. 77891

Titanium dioxide
INS 171
ADI "khng gii hn".
2. nh ngha
Titan dioxyd c sn xut bng x l sulfat hoc x l

clorid. Cc iu kin x l xc nh dng ca sn phm
cui cng (cu trc tinh th anatase hoc rutil).
Trong quy trnh sulfat, acid sulfuric c dng phn hy
ilmenit (FeTiO3) hoc ilmenit v x titan. Sau mt lot cc
bc tinh ch, titan dioxyd phn lp ln cui c ra
bng nc, nung, v nghin mn.
Trong quy trnh clorid, kh clo phn ng vi khong cha
titan di iu kin kh to thnh titan tetra clorid khan,
cht ny c tinh ch v chuyn thnh titan dioxyd bng
oxy ha c nhit trc tip hoc bng phn ng vi lung
hi nc trong pha hi. C th cho acid hydrocloric c
phn ng vi khong cha titan to thnh dung dch titan
tetraclorid, sau tinh ch v chuyn thnh titan dioxyd
bng thy phn. Titan dioxyd c lc, ra v nung.
Ch phm Titan dioxyd thng mi c th c bao mt
lng nh nhm oxyd v/hoc silic dioxyd nng cao
c tnh k thut ca sn phm.
M s C.A.S.
13463-67-7
Cng thc ha hc
TiO2
Khi lng phn t
79,88
3. Cm quan
Bt trng ti hi ng
4. Chc nng
Phm mu
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc, acid hydrocloric, acid sulfuric long

v cc dung mi hu c. Ha tan chm trong acid
hydrofluoric v acid sulfuric c nng.
Phn ng mu
5.2. tinh khit

Mt khi lng khi sy
Khng c qu 0,5%. Sy 105 C trong 3 gi.
Mt khi lng khi nung
Khng c qu 1,0% so vi ch phm kh.

Nung 800 C
Nhm oxyd v/ hoc
Khng c qu 2,0%, mt cht hoc c hai cht
silic dioxyd
Cc cht tan trong acid
Khng c qu 0,5%; Khng c qu 1,5% i vi ch

phm cha nhm oxyd hoc silic dioxyd.
Cc cht tan trong nc
Khng c qu 0,5%.
Cc tp cht tan trong

acid hydrocloric 0,5 N
Stibi
Khng c qu 2,0 mg/kg
Arsen
Khng c qu 1,0 mg/kg
Cadmi
Khng c qu 1,0 mg/kg
Ch
Thy ngn
Khng c qu 1,0 mg/kg
5.3. nh lng
Khng c nh hn 99,0 % tnh theo ch phm kh

(khng tnh nhm oxyd v silic dioxyd)
6. Phng php th
6.1 nh tnh
Phn ng mu
Thm 5 ml acid sulfuric vo 0,5 g mu th, un nng nh

ti khi xut hin khi ca acid sulfuric, ri ngui. Pha
long cn thn ti khong 100 ml vi nc v lc. Thm
vi git hydrogen peroxyd vo 5 ml dch lc trong; xut
hin ngay mu cam.
6.2. tinh khit

Cc cht tan trong acid
To hn dch 5 g mu th trong 100 ml acid hydrocloric 0,5

N v un trn ni cch thy si 30 pht, thnh thong
khuy. Lc qua phu lc s vi giy lc si thy tinh. Ra
ba ln, mi ln bng 10 ml acid hydrocloric 0,5 N, gp dch
lc v dch ra, c cn ri nung nng ti ti khi lng
khng i.
Tp cht tan trong nc
Tin hnh theo ch dn phn cc cht tan trong acid (bn

trn), dng nc thay cho acid hydrocloric 0,5 N.
Cc tp cht tan trong

acid hydrocloric 0,5 N
Antimon, arsen, cadmi v
ch
Th theo hng dn ti JECFA monograph 1-Vol. 4: cc

phng php th chung.
Chuyn 10,0 g mu th vo cc 250 ml, thm 50 ml acid
hydrocloric 0,5 N, y bng mt knh ng h v un ti
si trn bp in phng (bp in kn). un si nh trong
15 pht, chuyn dch treo vo chai ly tm 100 ti 150 ml, ly

tm 10 ti 15 pht, hoc ti khi cc cht khng tan lng
xung. Gn dch chit lp trn qua giy lc Whatman No.
4 hoc tng ng, thu dch lc vo bnh nh mc 100
ml, c gng gi li trong l c phn khng tan cng
nhiu cng tt. Thm 10 ml nc nng vo cc ban u,
ra mt knh ng h bng nc, v cho vo chai ly tm.
Dng a thy tinh khuy to hn dch ri ly tm. Gn
qua cng t giy lc trn, thu dch ra vo bnh nh mc
cha dch chit ban u. Lm lp li qu trnh ra hai ln
na. Cui cng ra giy lc bng 10 ti 15 ml nc nng.
ngui hn hp trong bnh ti nhit phng, pha long
ti vch bng nc, lc u.
Xc nh antimon, cadimi v ch, dng k thut AAS/ICPAES ph hp vi mc qui nh. Xc nh arsen dng k
thut ICP-AES/AAS hydrid ha. C th thay th bng cch
xc nh arsen dng Phng php II ca Php th gii
hn Arsen, ly 3 g mu th thay cho 1 g. La chn c mu
th c th da trn nguyn tc ca phng php m t
trong JECFA monograph 1-Vol. 4: cc phgn php th
chung.
Thy ngn
Nhm oxyd
Xc nh bng k thut quang ph hp th nguyn t ha

hi lnh. Chn la c mu thch hp vi nng quy nh.
th theo hng dn ti JECFA monograph 1-Vol. 4
Thuc th v dung dch mu th
Km sulfat 0,01 N
Ha tan 2,9 g km sulfat (ZnSO47H2O) trong nc va
1000 ml. Chun ha dung dch theo cch sau: Cn chnh
xc 500 mg si nhm c tinh khit cao (99,9%), ha tan
trong 20 ml acid hydrocloric c, un nng nh ti khi tan
ht, sau chuyn sang bnh nh mc 1000 ml, thm
nc ti vch, lc u. Chuyn 10 ml dung dch ny sang
bnh nn 500 ml cha 90 ml nc v 3 ml acid hydrocloric
c, thm 1 git da cam methyl (TT) v 25 ml dinatri
ethylen diamin tetraacetat (EDTA) 0,02 M. Thm tng git
dung dch amoniac 1/5 ti khi mu va chuyn hon ton
t mu thnh mu vng cam. Sau , thm:
(a): 10 ml dung dch m amoni acetat (77 g amoni
acetat thm 10 ml acid acetic bng, thm nc ti va
1000 ml) v
(b): 10 ml dung dch diamoni hydrogen phosphat (150
g diamoni hydrogen phosphat trong 700 ml nc, iu
chnh ti pH 5,5 bng dung dch acid hydrocloric ,
thm nc ti va 1000 ml).
un si dung dch ny trong 5 pht, lm ngui nhanh ti
nhit phng di vi nc, thm 3 git da cam xylenol
(TT), lc u. Dng dung dch km sulfat lm dung dch
chun, chun ti im kt thc khi dung dch bt u
chuyn sang mu nu vng hoc mu hng bn trong 5-10
giy. (Ch : Phi chun nhanh khi gn im kt thc
bng cch thm nhanh tng lng 0,2 ml dung dch chun
ti khi bt u c s chuyn mu; mc d mu s b

nht trong 5-10 giy, l im kt thc thc. Khng quan
st c ln chuyn mu u tin s dn ti php chun
khng chnh xc. im kt thc nht mu khng xy ra
im kt thc th hai.)
Thm 2 g natri fluorid, un si hn hp trong 2-5 pht, lm
ngui di vi nc. Dng dung dch km sulfat chun
dung dch ny ti im kt thc khi dung dch bt u
chuyn sang mu nu vng hoc mu hng khng bn
nh m t trn.
Tnh chun T ca dung dch km sulfat theo cng thc
sau:
T = 18,896 m / V
Trong :
T l khi lng (mg) ca Al2O3 tc dng va vi 1 ml
dung dch km sulfat
m l khi lng (g) ca si nhm
V l s ml ca dung dch km sulfat dng ht im chun
th hai
18,896 = (R 1000 mg/g 10 ml/2)/1000 ml v
R l t s trng lng theo cng thc ca nhm oxyd v
ca nhm nguyn t
Dung dch mu th A
Cn chnh xc 1 g mu th v cho vo bnh nn thy tinh
giu silica 250 ml. Thm 10 g natri bisulfat (NaHSO4H2O).
(Ch : Khng c dng natri bisulfat nhiu hn qui nh,
nu d mui s nh hng ti php chun bng EDTA
phn sau trong quy trnh ny). Bt u un nng bnh
nhit thp trn bp in phng, sau tng nhit t
t ti khi nng ht c.
(Thn trng: thc hin qui trnh ny ni thng gi tt).
Khi hn hp ngng n lp bp, v xut hin khi nh ca
SO3, un nng trn ngn la ln bng u t Meeker,
nghing bnh sao cho hn hp mu th v natri bisulfat c
c li mt bn ca y ca bnh. Lc xoy lin tc ti
khi khi cht chy trong ra (tr silic dioxyd) nhng ch
khng un lu qu trnh kt ta titan dioxyd. Lm ngui,
thm 25 ml dung dch acid sulfuric 1:2, v un nng ti khi
khi nho ha tan v thu c dung dch trong sut. Lm
ngui, v pha long ti 120 ml bng nc. Cho que khuy
t vo bnh.
Dung dch mu th B
Pha 200 ml dung dch natri hydroxyd nng xp x 6,25
M. Thm 65 ml dung dch ny vo Dung dch mu th A,
va thm va khuy bng my khuy t; cho 135 ml dung
dch kim cn li vo mt bnh nh mc 500 ml.
Va khuy lin tc, va thm t t hn hp mu th vo
dung dch kim trong bnh nh mc 500 ml; thm nc ti
vch, lc u. (Ch : Nu dng th nghim ti thi im
qu 2 gi, phi hn hp trong bnh nh mc vo trong
chai polyethylen). yn cho phn ln ta lng xung
(hoc ly tm 5 pht), sau lc lp dch pha trn qua giy
Silic dioxyd
lc rt mn. Dn nhn dch lc l Dung dch mu th B.

Dung dch mu th C
Chuyn 100 ml dung dch mu th B vo bnh nn 500 ml,
thm 1 git da cam methyl (TT), acid ha bng dung dch
acid hydrocloric , sau thm d khong 3 ml. Thm 25
ml dung dch dinatri EDTA 0,02 M, v lc u. [Ch : Nu
bit hm lng xp x ca Al2O3, tnh th tch ti u ca
dung dch EDTA s thm theo cng thc:
(4 % Al2O3) + 5].
Thm tng git dung dch amoniac 1/5 ti khi mu bt u
chuyn hon ton t thnh mu vng cam. Sau
thm 10 ml mi dung dch 1 v dung dch 2 (xem trn) v
un si trong 5 pht. Lm ngui nhanh ti nhit phng
di vi nc, thm 3 git da cam xylenol (TT), lc u.
Nu dung dch c mu tm , mu nu vng, hoc mu
hng, iu chnh pH ti 5,35,7 bng cch thm acid
acetic. Ti pH ny, mu hng cho bit cha thm dung
dch EDTA, trong trng hp ny, b dung dch ny i v
lp li cch tin hnh ny vi 100 ml Dung dch mu th B
khc, dng 50 ml thay cho 25 ml dung dch dinatri EDTA
0,02 M.
Tin hnh
Dng dung dch chun km sulfat chun , chun
Dung dch mu th C ti im kt thc th nht khi bt
u c mu vng nu hoc hng bn trong 5-10 giy.
(Quan trng: Xem ch phn Km sulfat 0,01 N). Ln
chun u ny cn n hn 8 ml dung dch chun ,
nhng php chun s chnh xc hn nu lng dung
dch chun dng l 10-15 ml.
Thm 2 g natri fluorid vo bnh chun , un si hn hp
trong 2-5 pht, v lm ngui di vi nc. Chun
dung dch ny bng dung dch km sulfat chun ha ti
khi c mu nu vng hoc hng khng bn nh m t
trn.
Tnh ton:
Tnh phn trm nhm oxyd (Al2O3) trong mu th theo
cng thc sau:
% Al2O3 = 100 (0,005VT)/m
Trong :
V l s ml dung dch km sulfat 0,01 N dng ht trong ln
chun th hai,
T l chun ca dung dch km sulfat,
m l khi lng (g) ca mu th, v
0,005 = 500 ml / (1000 mg/g 100 ml).
Cn chnh xc 1 g mu th v cho vo bnh nn thy tinh
giu silica 250 ml. Thm 10 g natri bisulfat (NaHSO4H2O).
un nng bng u t Meeker, va un va lc xoy
bnh ti khi hn hp phn hy v nng chy hon ton, v
tr nn trong sut, tr phn silic dioxyd, sau lm ngui.
(Thn trng: Khng c un qu nng hn hp trong
bnh ngay t u, v un cn thn trong sut qu trnh
chy khi bn ra ngoi).
Thm 25 ml dung dch acid sulfuric vo hn hp nng

chy ngui v un nng cn thn v t t ti khi hn
hp ny ha tan. ngui v thm cn thn 150 ml nc
bng cch rt tng lng nh vo theo thnh bnh, thnh
thong khuy trnh qu nng v b bn ra ngoi.
ngui hn hp trong bnh, lc qua giy lc khng tro mn,
dng phu lc 60o t chy. Ra ly ht ton b silic dioxyd
ra khi bnh, ln trn giy lc bng dung dch acid sulfuric
1/10. Cho giy lc v cn trn giy lc vo chn nung
platin, lm kh trong t sy 120 C, v lm t nng
chn nung c y kn mt phn bng ngn la Bunsen.
giy lc khng b chy thnh ngn la, trc tin t
nng t pha np, ri sau mi nung chn t pha di.
Khi giy lc tn ht, chuyn chn nung sang l nung v
nung 1000 C trong 30 pht. ngui trong bnh ht m,
v cn. Thm 2 git dung dch acid sulfuric v 5 ml dung
dch acid hydrofluoric c (t trng tng i khong 1,15),
ri c cn thn ti kh, u tin un nng nh trn bp
in phng ( loi HF), sau t bng ngn la Bunsen
( loi H2SO4). Lm cn thn trnh bn ra ngoi, c
bit sau khi loi HF. Nung 1000 C trong 10 pht,
ngui trong bnh ht m, v cn. chnh lch gia hai
ln cn l lng SiO2 c trong mu th.
6.3. nh lng
Cn chnh xc khong 150 mg mu th trc sy
kh 105 C trong 3 gi, cho vo bnh nn 500 ml. Thm
5 ml nc v lc ti khi thu c hn dch ging sa ng
nht. Thm 30 ml dung dch acid sulfuric v 12 g amoni
sulfat, lc u. Lc u un nh, sau un mnh ti khi
thu c dung dch trong sut. ngui, sau pha long
cn thn vi 120 ml nc v 40 ml dung dch acid
hydrocloric, khuy. Thm 3 g nhm kim loi, y ngay bnh
bng nt cao su c gn ng thy tinh hnh ch U, u kia
ca ng ch U ngp trong dung dch natri bicarbonat bo
ha ng trong l rng ming 500 ml, v to kh hydro.
yn trong vi pht sau khi nhm kim loi ha tan hon ton
to thnh dung dch trong sut mu tm . Lm ngui
xung di 50 C di vi nc, tho nt cao su cng ng
ch U. Thm 3 ml dung dch kali thiocyanat bo ha lm
ch th, v chun ngay bng dung dch st (III) amoni
sulfat 0,2 N ti khi thu c dung dch c mu nu nht
bn trong 30 giy. Tin hnh lm mt mu trng v hiu
chnh nu cn thit.
1 ml dung dch st (III) amoni sulfat 0,2 N tng ng vi
7,990 mg TiO2.
Ph lc 29
YU CU K THUT V PHNG PHP TH I VI ST OXYD
1. Tn khc, ch s
Iron oxides;
St oxyd vng: CI Pigment yellow 42 and 43; CI(1975) No.
77492; INS 172(iii)
St oxyd : CI Pigment red 101 and 102; CI (1975)
No.77491; INS 172(ii)
St oxyd en: CI Pigment black 11; CI (1975) No.77499;
INS 172(i)
ADI= 0 - 0,5 mg/kg th trng.
2. nh ngha
Ch phm c sn xut t st (II) sulfat bng qu trnh

ngm, nung nng, loi nc, phn hu, ra, lc, sy,
nghin.
Ch phm cha cc thnh phn st oxyd khan hoc dng
hydrat, c cc mu t vng, n nu v en. Cht lng
st oxyd dng trong thc phm phn bit vi st oxyd k
thut qua hm lng cc kim loi khc trong ch phm do
vic la chn v kim sot ngun nguyn liu st v cng
on tinh ch ho hc trong sut qu trnh sn xut.
Tn ha hc
St oxyd vng: St oxyd dng hydrat, St (III) oxyd dng

hydrat
St oxyd : St sesquioxyd, st (III) oxyd khan
St oxyd en: hp cht st (II) st (III) oxyd, st (II, III) oxyd
M s C.A.S.
St oxyd vng: 51274-00-1

St oxyd : 1309-37-1
St oxyd en: 1317-61-9
Cng thc ha hc
St oxyd vng: FeO(OH) . xH2O

St oxyd : Fe2O3
St oxyd en: FeO . Fe2O3
Khi lng phn t
88,85 FeO(OH)
159,70 Fe2O3
231,55 FeO . Fe2O3
3. Cm quan
Bt mu en, nu, hoc vng
4. Chc nng
Phm mu
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc v cc dung mi hu c; tan trong

cc acid v c m c.
Hp cht ho tan trong

nc
Khng c qu 1,0%.
5.2. tinh khit

Gim khi lng khi
sy kh
St oxyd : khng c qu 1,0% (sy nhit 105oC

trong 4h).
Arsen
Cadmi
Ch
Thu ngn
Khng c qu 1,0 mg/kg
5.3. Hm lng
Hm lng st khng c thp hn 60,0%.
6. Phng php th
6.1. tinh khit
Arsen, cadmi v ch
Cn 5 g mu th v chuyn vo mt cc thu tinh. Cho

thm 50 ml dung dch acid sulfuric c v lm nng trn
mt a nng cho n khi ho tan hon ton. Pha long
bng nc ct n vch 100 ml trong bnh nh mc. Xc
nh cadmi v ch bng k thut hp th nguyn t thch
hp cho hm lng quy nh v arsen da trn Phng
php II ca JECFA monograph 1-Vol. 4.
Thu ngn

- Xc nh bng k thut hp th nguyn t thch hp cho
m t trong chuyn lun cc phng php phn tch cng
c.
6.2. nh lng
Cn 0,2 g mu th, chnh xc n mg, cho thm 10 ml dung
dch acid hydrocloric 5N v cn thn un n si trong mt
bnh nn 200 ml cho ti khi mu th tan hon ton.
ngui, cho thm 6 n 7 git dung dch hydrogen peroxyd
30% v tip tc cn thn un n si cho n khi ton b
hydrogen peroxyd d b phn hu (khong 2 n 3 pht).
ngui, cho thm 30 ml nc ct v khong 2 g kali iodid
v yn trong 5 pht. Cho thm 30 ml nc ct v chun
vi dung dch natri thiosulfat 0,1N, thm ch th l dung
dch h tinh bt (TT) n im tng ng th kt thc
chun . Mi ml dung dch natri thiosulfat 0,1N tng
ng vi 5,585 mg Fe (III).
Ph lc 30
YU CU K THUT V PHNG PHP TH I VI FAST GREEN FCF
1. Tn khc, ch s
CI food green 3,CI (1975): 42053

INSL: 143
2. nh ngha
Ch yu gm dinatri 3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3sulfonbenzyl)amino]phenyl](4-hydroxy-2sulfonphenyl)methylen]-2,5-cyclohexadien-1yliden]amoniomethyl]benzensulfon v cc ng phn ca

n cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4
l cc thnh phn khng mu chnh.
Tn ha hc
Dinatri 3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3-sulfonbenzyl)
amino] phenyl](4-hydroxy-2-sulfonphenyl)methylen]-2,5cyclohexadien-1-yliden] amoniomethyl]-benzensulfon;
Mui dinatri ni ca N-ethyl-N-[4[[4-ethyl[(3sulfophenyl)methyl]amino]phenyl](4-hydroxy-2-sulfophenyl)
methylen]-2,5-cyclohexadien-1-yliden]-3-sulfobenzenmethanamini hydroxyd.
M s C.A.S.
2353-43-9
Cng thc ha hc
C37H34N2Na2O10S3
Cng thc cu to
CH2CH3
N CH2
HO
SO3Na
H3CH2C
CH2
SO3
Khi lng phn t
808,86
3. Cm quan
Dng bt hoc tinh th mu n tm nu.
4. Chc nng
Phm mu.
SO3Na
5. Yu cu k thut
5.1. nh tnh
tan
nh tnh cc cht mu
5.2. tinh khit

Gim khi lng khi lm
kh ti 135oC

mui natri.
Cht khng tan trong

nc
Khng c qu 0,2%
Ch
Crom
Cc cht mu ph
Khng c qu 6,0%.
Cc cht hu c ngoi
cht mu
Khng c qu 0,5% tng cc acid 2-; 3-; v 4Formylbenzensulfonic v mui natri ca cc acid ny.
Khng c qu 0,3% tng cc acid 3- v 4-[N-Ethyl-N-(4sulfophenyl)amino]methylbenzensulfonic v mui dinatri
ca cc acid ny.
Khng c qu 0,5% tng ca acid 2-Formyl-5hydroxybenzensulfonic v mui natri ca n.

khng sulfon ho
Leuco base
Khng c qu 5,0%.
Cc cht c th chit
bng ether
Khng c qu 0,4%.
5.3. Hm lng
6. Phng php th
6.1. tinh khit
Ch

m t trong JECFA monograph 1-Vol. 4 phn cc phng
php phn tch cng c.
Cc cht mu ph
Pha dung dch mu th 1%. Dng syring chm dung dch

mu th ln bn mng silicagel (20 x 20 cm) sao cho lng
chm l 1 mg mu th. bn mng kh trong ch ti
khong 20 pht sau cho bn mng vo bnh chy sc k
bo ha h dung mi khai trin sc k: acetonitril/alcol
isoamylic/methyl ethyl keton/nc/dung dch amoniac vi t
l theo th tch l 50/50/15/10/5. khai trin n khi tuyn
dung mi gn vi nh ca bn mng. Ly bn mng ra v
kh. Cht mu ph xut hin ti cc v tr sau: trn cng

l 2 vt ca cht mu ph sulfon ha thp, tip theo l vt
chnh ng mu ca xanh lc bn FCF gn y bn mng.
Co phn silicagel pha trn vt chnh ng mu. Chit
mu ra khi silicagel bng ethanol, lc qua phu lc thy
tinh xp v o quang ph. hp th ring tiu chun ca
cc cht mu ph l 0,126 mgL-1cm-1.
Ch : Trong qu trnh tnh cc cht mu ph, tha nhn
rng hp th ring ti cc cc i hp th ca chng
tng t nh hp th ring ti cc i hp th ca cc
cht gc. Dung dch chun phi c o trong thi gian 1
gi k t lc pha. Dch chit cht mu ph (ra khi silicagel)
cn c o quang ph cng nhanh cng tt.
Cc cht hu c ngoi
cht mu

- S dng k thut sc k ct, vi cc iu kin hp th (v
d) nh sau:
Acid 3-formylbenzensulfonic: 0,495 mg L-1 cm-1 ti 246 nm
trong HCl long
Acid 3-[(ethyl)(4-sulfophenyl)amino] methyl benzensulfonic:
0,078 mg L-1 cm-1 ti 277 nm trong dung dch amoniac
long
Acid 2-formyl-5-hydroxybenzensulfonic: 0,080 mg L-1 cm-1
ti 335 trong dung dch amoniac long.
Leuco base

- Cn chnh xc 110 5 mg mu v tin hnh theo hng
dn trong chuyn lun th Leuco base trong cht mu
Triarylmethansulfon ha.
hp th ring (a)=0,6 mgL-1cm-1 ti bc sng khong
625 nm. T s = 0,971.
6.2. nh lng

Cn 1,9 - 2,0 g mu th;
mu (D).

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CNG HA X HI CH NGHA VIT NAM

QCVN 4-10 : 2010/BYT

QUY CHUN K THUT QUC GIA

QUY CHUN K THUT QUC GIA

Quy nh k thut v phng php th i vi

Quy nh k thut v phng php th i vi

Quy nh k thut v phng php th i vi quinolin

Quy nh k thut v phng php th i vi sunset

Quy nh k thut v phng php th i vi

Quy nh k thut v phng php th i vi cao v

2. Cc yu cu k thut quy nh trong Quy chun ny c th theo JECFA

V. T CHC THC HIN

Vng ngh, Kurkum

Curcumin c sn xut bng cch chit c ngh - thn

Cc thnh phn mu chnh:

R1= OCH3; R2=H

Khi lng phn t

Bt tinh th mu vng cam.

Khng tan trong nc v diethyl ether; tan trong ethanol

Dung dch mu th trong ethanol c mu vng v cho

t yu cu (m t trong phn Phng php th).

5.2. tinh khit

Khng c qu 30,0 mg/kg

Khng c qu 25,0 mg/kg

Khng c qu 50,0 mg/kg

Khng c qu 2,0 mg/kg

Khng nh hn 90,0% tng cc thnh phn mu.

X l dung dch mu th trong nc hoc trong ethanol

Chm 5 L dch th (0,01 g mu th trong 1 ml ethanol

dch amoniac (4:4:2:1), dung mi khai trin trn bn

Xc nh bng k thut hp th nguyn t thch hp cho

Vitamin B2, Lactoflavin

Riboflavin; 3,10-dihydro-7,8-dimethyl-10-[(2S,3S,4R)2,3,4,5-tetrahydroxypentyl]benzo-[g]pteridin-2,4-dion; 7,8dimethyl-10-(1'-Dribityl) isoalloxazin.

Khi lng phn t

Bt tinh th mu vng n vng cam, c mi nh.

Rt kh tan trong nc, thc t khng tan trong cn, ether,

S dng dung dch mu th trong nc, chun b trong

[ ]20,D: trong khong -115o v -140o.

t yu cu (m t trong phn Phng php th).

5.2. tinh khit

t yu cu (m t trong phn Phng php th).

Cc amin thm bc nht

Khng c qu 100,0 mg/kg tnh theo anilin.

Khng c qu 2,0 mg/kg.

Sy kh mu ti 100 oC trong 4 g. Cn 50 mg v ha tan

6.2. tinh khit

Sy ti 105 oC, trong 4 gi

- Th theo JECFA monograph 1 - vol.4

Chun b dung dch chun khng c lumiflavin cho th

Th theo hng dn ti JECFA monograph 1 - vol.4. S

CI Food Yellow 4, FD&C Yellow No. 5; S CI (1975): 19140

Ch yu gm Trinatri 5-hydroxy-1-(4-sulfonphenyl)-4-(4sulfonphenylazo)-H-pyrazol-3-carboxylat cng cc cht

Khi lng phn t

Dng bt hoc ht mu cam nht.

Tan trong nc, t tan trong ethanol.

Phi c phn ng c trng ca cc cht mu

5.2. tinh khit

Khng c qu 15,0% cng vi clorid v sulfat tnh theo

Cht khng tan trong

Khng c qu 2,0 mg/kg.

Khng c qu 0,5% (tng cc acid Tetrahydroxysuccinic;

Cc amin thm bc nht Khng c qu 0,01% tnh theo anilin.

Khng c thp hn 85,0% tng cc cht mu.