Coeus.vn a4.CHẤT LÀM DÀY

CNG HO X HI CH NGHA VIT NAM
QCVN 4 - 21: 2011/BYT
QUY CHUN K THUT QUC GIA

V PH GIA THC PHM - CHT LM DY
National technical regulation on Food Additive Thickeners
H NI - 2011
QCVN 4 - 21: 2011/BYT
Li ni u
QCVN 4-21:2011/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 01/2011/TT-BYT ngy 13 thng 01
nm 2011 ca B trng B Y t.
QCVN 4 - 21: 2011/BYT
QUY CHUN K THUT QUC GIA

V PH GIA THC PHM - CHT LM DY
National technical regulation on Food Additive Thickeners
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 cht lm dy 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
cht lm dy 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. Cht lm dy: l ph gia thc phm c s dng lm tng nht
ca thc phm.
3.2. JECFA monograph 1 - Vol. 4 (JECFA monographs 1 - Combined
compendium of food additive 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.
QCVN 4 - 21: 2011/BYT

II. YU CU K THUT, PHNG PHP TH V LY MU
1. Yu cu k thut v phng php th i vi cc cht lm dy c quy
nh ti cc ph lc ban hnh km theo Quy chun ny nh sau:
1.1.
Ph lc 1:
Yu cu k thut v phng php th i vi acid

alginic
1.2.
Ph lc 2:
Yu cu k thut v phng php th i vi kali

alginat
1.3.
Ph lc 3 :
Yu cu k thut v phng php th i vi amoni

alginat
1.4.
Ph lc 4:
Yu cu k thut v phng php th i vi calci

alginat
1.5.
Ph lc 5:
Yu cu k thut v phng php th i vi propylen

glycol alginat
1.6.
Ph lc 6:
Yu cu k thut v phng php th i vi agar
1.7.
Ph lc 7:
Yu cu k thut v phng php th i vi

carrageenan v mui Na, K, NH4 ca n
1.8.
Ph lc 8:
Yu cu k thut v phng php th i vi gm u

Carob
1.9.
Ph lc 9:
Yu cu k thut v phng php th i vi gm

guar
1.10.
Ph lc 10:

tragacanth
1.11.
Ph lc 11:

arabic
1.12.
Ph lc 12:

xanthan
1.13.
Ph lc 13:

karaya
1.14.
Ph lc 14:

tara
1.15.
Ph lc 15:

gellan
1.16.
Ph lc 16:
Yu cu k thut v phng php th i vi pectin
1.17.
Ph lc 17:
Yu cu k thut v phng php th i vi methyl

celulose
1.18.
Ph lc 18:
Yu cu k thut v phng php th i vi methyl

ethyl celulose
1.19.
Ph lc 19:
Yu cu k thut v phng php th i vi natri

carboxymethyl celulose
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1.20.
Ph lc 20:
Yu cu k thut v phng php th i vi gelatin

thc phm
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 c gi tr 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 cht lm dy 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 cht lm dy
Vic kim tra cht lng, v sinh an ton i vi cc cht lm dy 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 cht lm dy 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.
QCVN 4 - 21: 2011/BYT

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.
QCVN 4 - 21: 2011/BYT

Ph lc 1
YU CU K THUT V PHNG PHP TH
I VI ACID ALGINIC
1. Tn khc, ch s
Alginic acid
INS 400
ADI "Khng gii hn"
2. nh ngha
Acid alginic t nhin thu c bng cch thy phn keo

polysaccarid t cc loi rong bin nu (Phaeophyceae). N
l mt polymer mch thng gm ch yu cc monomer ca
acid -1,4-D-mannuroric v acid a-1,4-L-glucuronic. Cc
monomer ny thng c sp xp thnh cc khi
homopolymer cch nhau bi cc vng 2 monomer acid tun
t xen k nhau.
Ch s C.A.S.
9005-32-7
Cng thc ha hc
(C6H8O6)n
Cng thc cu to
Cng thc cu to do Phillips, Wedlock and Williams cng

b trong Gms and Stabilizers for the Food Industry 5
(1990), nh xut bn Oxford University Press.
Khi lng phn t
S lng v th t ca cc phn mannuronat v gluconat c

th thay i trong alginat t nhin. Cng thc trn cha th
hin s kt hp vi nc.
n v cu trc : 176,13 (l thuyt); 200 (trung bnh thc t).
i phn t: 10.000-600.000 (trung bnh in hnh)
3. Cm quan
Dng ht, bt hoc si mnh mu trng n vng nu.
4. Chc nng
Cht n nh, cht lm dy, cht to gel, cht nh ha
5. Yu cu k thut
5.1. nh tnh
pH
Huyn ph mu th 0,3/10 c pH trong khong 2,0 - 3,5.
To kt ta vi amoni
sulfat
Phi c phn ng to kt ta vi amoni sulfat c trng.
Alginat
Phi c phn ng c trng ca alginat.
5.2. tinh khit

7
QCVN 4 - 21: 2011/BYT

Gim khi lng khi sy
kh
Khng c qu 15%
(Sy ti 105o trong 4 gi).
Tro sulfat
Khng c qu 8% tnh theo ch phm lm kh.
Cht khng tan trong

natri hydroxyd
Arsen
Khng c qu 3,0 mg/kg.
Ch
5.3 Cc yu cu v vi
sinh vt
Tng s vi sinh vt hiu
kh
Khng c qu 5000 khun lc/g. Ban u chun b m

pha long 10-1 bng cch thm 50g mu vo 450ml nc
pha long m phosphat Butterfield v ng ha bng my
nghin cao tc.
Tng s nm men-mc
Khng c qu 500 khun lc/g
Coliforms
m tnh
Salmonella
m tnh
5.4. Hm lng
(C6H8O6)n
Ch phm khan cha khng thp hn 20,0% v khng c

qu 23,0% CO2 tng ng vi khng thp hn 91,0% v
khng c qu 104,5% acid alginic (C6H8O6)n.
6. Phng php th
6.1. nh tnh
To kt ta vi amoni
sulfat
Thm dung dch amoni sulfat bo ha vo dung dch mu

th 0,5% trong dung dch natri hydroxyd (TS), vi th tch
bng 1/2 th tch dch th. Khng c c kt ta. Php th
ny phn bit acid alginic vi thch, natri carboxymethyl
cellulose, carragenan, gelatin, gm carob bean, methyl
celllulose, pectin de-ester ha, tinh bt.
Alginat
Ha tan 0,1 g mu th vi 0,15 ml dung dch natri hydroxyd

0,1 N. Thm 1 ml dung dch st (III) sulfat (TS). Trong 5 pht,
mu anh o s xut hin v m dn thnh mu tm
sm.
6.2. tinh khit

Cht khng tan trong
natri hydroxyd
Cn khong 1 g (chnh xc n mg) mu th, ha tan trong

100 ml dung dch natri hydroxyd (TS), ly tm, gn ly phn
khng tan. Ra cn 5 ln bng nc, ly tm hoc gn b
nc ra. Dng nc chuyn phn khng tan vo phu lc
thy tinh cn b, sy ti 105o trong 1 gi, ngui v cn.
Tnh hm lng % theo khi lng ch phm c lm
kh.
8
QCVN 4 - 21: 2011/BYT

Arsen
Th theo JECFA monograph 1 - Vol.4. (phng php II)
Ch
- Th theo JECFA monograph 1 - Vol.4.

- Xc nh bng k thut quang ph 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
Tin hnh theo hng dn trong chuyn lun xc nh
carbon dioxyd bng phng php decarboxyl ha ti JECFA
monograph 1 - Vol.4.
Mi ml natri hydroxyd 0,25 N s dng tng ng vi 5,5
mg carbon dioxyd hoc 25 mg acid alginic (tng ng
khi lng 200).
QCVN 4 - 21: 2011/BYT

Ph lc 2
I VI KALI ALGINAT
1. Tn khc, ch s
INS 402
ADI "Khng gii hn"
2. nh ngha
Mui Kali ca acid alginic
Ch s C.A.S.
9005-36-1
Cng thc ha hc
(C6H7KO6)n
Cng thc cu to


hin s kt hp vi nc.
Khi lng phn t

i phn t: 10000-600000 (trung bnh in hnh)
3. Cm quan
4. Chc nng
5. Yu cu k thut
5.1. nh tnh
tan
Tan chm trong nc, to ra dung dch nht. Khng tan

trong ethanol v ether.
To kt ta vi calci
clorid
Phi c phn ng to kt ta vi calci clorid c trng.
To kt ta vi amoni
sulfat
Phi c phn ng to kt ta vi amoni sulfat c trung.
Alginat
Kali
Phi c phn ng c trng ca kali.
5.2. tinh khit

Gim khi lng khi sy
Khng c qu 15% (Sy ti 105o trong 4 gi).

10
QCVN 4 - 21: 2011/BYT

kh
Cht khng tan trong
nc
Arsen
Ch
Khng c qu 5,0 mg/kg
5.3. Cc yu cu v vi
sinh vt
kh

pha long m phosphat Butterfield v ng nht ha bng
my nghin cao tc.
Tng s nm men-mc
Coliforms
m tnh
Salmonella
m tnh
5.4. Hm lng
(C6H7KO6)n
Ch phm khan cha khng thp hn 16,5% v khng c

qu 19,5% CO2 tng ng vi khng thp hn 89,2% v
khng c qu 105,5% kali alginat (C6H7KO6)n.
6. Phng php th
6.1. nh tnh
To kt ta vi calci
clorid
Thm dung dch calci clorid 2,5% vo dch th l dung dch

mu th 0,5% trong dung dch natri hydroxyd (TS), vi th
tch bng 1/5 th tch dch th. Xut hin ta khi ln dng
keo. Php th ny phn bit calci alginat vi gm arabic,
natri carboxymethyl cellulose, carragenan, gelatin, gm
ghatti, gm karaya, gm carob bean, methyl celllulo, gm
tragacanth.
To kt ta vi amoni
sulfat
Thm dung dch amoni sulfat bo ha vo dch th l dung

dch mu th 0,5% trong dung dch natri hydroxyd (TS), vi
th tch bng 1/2 th tch dch th. Khng c xut hin
ta. Php th ny phn bit calci alginat vi thch, natri
carboxymethyl cellulose, carragenan, pectin de-este ha,
gelatin, gm carob bean, methyl celllulo v tinh bt.
Alginat
Th theo hng dn ti JECFA monograph 1 - Vol.4.
Kali
6.2. tinh khit

Cht khng tan trong
nc
Cn 2 g (chnh xc n 0,1 mg) mu, phn tn vo 800 ml

nc trong bnh 2000 ml. Trung ha ti pH 7 bng dung dch
natri hydroxyd (TS) v thm d 3 ml. Thm 40 ml dung dch
hydrogen peroxyd 30% (kl/kl), y bnh v un trong khong
11
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Arsen
Ch
1 gi, khuy thng xuyn. Lc nng qua mt chn lc

Gooch c mng lc bng x thy tinh cn b (2,4 cm, No
934 AH, Reeve Angel & Co, Clifton, N.Y., USA hoc tng
ng). Nu qu trnh lc b chm do dung dch mu c
nht cao, un si n khi nht gim n mc c th lc
c. Ra chn lc bng nc nng, sy chn lc v phn
khng tan ti 105o trong 1 gi, ngui v cn. Tnh hm
lng % ca phn khng tan sy kh.
Th theo hng dn ti JECFA monograph 1 - Vol.4
(phng php II).
6.3. Cc yu cu v vi
sinh vt
6.4. nh lng
mg carbon dioxyd hoc 29,75 mg kali alginat (tng ng
khi lng 238).
12
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Ph lc 3
I VI AMONI ALGINAT
1. Tn khc, ch s
INS 403
ADI "Khng gii hn"
2. nh ngha
Mui Amoni ca acid alginic
Ch s C.A.S.
9005-34-9
Cng thc ha hc
(C6H11NO6)n
Cng thc cu to


hin s kt hp vi nc.
Khi lng phn t

3. Cm quan
4. Chc nng
5. Yu cu k thut
5.1. nh tnh
tan
Tan chm trong nc, to ra dung dch nht. Khng tan

trong ethanol v ether.
To kt ta vi calci
clorid
To kt ta vi amoni
sulfat
Phi c phn ng to kt qu vi amoni sulfat c trng.
Alginat
Amoni
Phi c phn ng c trng ca amoni.
5.2. tinh khit
13
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Gim khi lng khi sy
kh
Cht khng tan trong

nc
Tro sulfat
Khng c qu 7,0% tnh theo ch phm lm kh.
Ch
5.3 Cc yu cu v vi
sinh vt
kh

my nghin cao tc.
Tng s nm men-mc
Coliforms
m tnh
Salmonella
m tnh
5.4. Hm lng
(C6H11NO6)n
Ch phm lm kh cha khng thp hn 18% v khng

c qu 21% CO2 tng ng vi khng thp hn 88,7%
v khng c qu 103,6% amoni alginat (C6H11NO6)n.
6. Phng php th
6.1. nh tnh
To kt ta vi calci
clorid

tragacanth.
To kt ta vi amoni
sulfat

Alginat

0,1 N, lc k mu th tan cng nhiu cng tt. Thm 1 ml
dung dch st(III) sulfat (TS). Trong 5 pht, mu hng anh
o s xut hin v m dn thnh mu tm sm.
Amoni
6.2. tinh khit
14
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Cht khng tan trong
nc

nc trong bnh dung tch 2000 ml. Trung ha ti pH 7 bng
dung dch natri hydroxyd (TS) v thm d 3 ml. Thm 40 ml
dung dch hydrogen peroxyd 30% (kl/kl), y bnh v un
trong khong 1 gi, khuy thng xuyn. Lc nng qua mt
chn lc Gooch c mng lc bng x thy tinh cn b (2,4
cm, No 934 AH, Reeve Angel & Co, Clifton, N.Y., USA hoc
tng ng). Nu qu trnh lc b chm do dung dch mu
c nht cao, un si n khi nht gim n mc c
th lc c. Ra chn lc bng nc nng, sy chn lc
v phn khng tan ti 105o trong 1 gi, ngui v cn.
Tnh hm lng % ca phn cn khng tan sy kh.
Ch

6.3. nh lng
mg carbon dioxyd.
15
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Ph lc 4
I VI CALCI ALGINAT
1. Tn khc, ch s
INS 404
ADI "Khng gii hn"
2. nh ngha
Mui calci ca acid alginic
Ch s C.A.S.
9005-35-0
Cng thc ha hc
(C6H7Ca1/2O6)n
Cng thc cu to
Cng thc cu to do Phillips, Wedlock and Williams cng b

trong Gms and Stabilizers for the Food Industry 5 (1990), nh
xut bn Oxford University Press.

hin s kt hp vi nc.
Khi lng phn t

i phn t: 10.000-600.000 (trung bnh in hnh)
3. Cm quan
4. Chc nng
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc v ether. t tan trong ethanol, tan

chm trong dung dch natri polyphosphat, natri carbonat v
cc cht kt hp vi ion calci
To kt ta vi calci
clorid
To kt ta vi amoni
sulfat
Phi c phn ng to kt ta vi amoni sulfat c trng.
Alginat
Calci
Phi c phn ng c trng ca calci.
5.2. tinh khit

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Gim khi lng khi
sy kh
Arsen

(th theo hng dn ti JECFA monograph 1 - Vol.4 phng php II).
Ch
5.3. Cc yu cu v vi
sinh vt
Tng s vi sinh vt
hiu kh

my nghin cao tc.
Tng s nm men-mc Khng c qu 500 khun lc/g

Coliforms
m tnh
Salmonella
m tnh
5.4. Hm lng
(C6H7Ca1/2O6)n
Khng thp hn 18,0% v khng c qu 21,0% CO2

tng ng vi khng thp hn 89,6% v khng c qu
104,5% calci alginat (C6H7Ca1/2O6)n tnh theo ch phm
khan.
6. Phng php th
6.1. nh tnh
To kt ta vi calci
clorid

tragacanth.
To kt ta vi amoni
sulfat

Alginat

0,1 N, lc k mu th tan cng nhiu cng tt. Thm 1 ml
dung dch st(III) sulfat (TS). Trong 5 pht, mu hng anh
o s xut hin v m dn thnh mu tm sm.
Calci
6.2. tinh khit
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Arsen

(phng php II).
Ch

6.3. nh lng
mg carbon dioxyd hoc 27,38 mg calci alginat (tng ng
khi lng 219).
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Ph lc 5
I VI PROPYLEN GLYCOL ALGINAT
1. Tn khc, ch s
Ester ca acid alginic vi 1,2-propan-diol; Hydroxylpropyl

alginat; Propan-1,2-diol alginat
INS 405
ADI "khng gii hn"
2. nh ngha
L ester ca acid alginic, trong mt s nhm carboxyl b

ester ha vi propylen glycol, mt s khc trung ha vi kim
cn li l t do.
Ch s C.A.S.
9005-37-2
Cng thc ha hc
(C9H14O7)n ( ester ha)
Khi lng phn t
n v cu trc : 234,21 (l thuyt).

3. Cm quan
4. Chc nng
Cht n nh, cht lm dy, cht nh ha.
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, to ra dung dch nht, keo. Tan trong hn

hp dung mi ethanol/nc n 60% ty thuc mc ester
ha.
To kt ta vi acid
sulfuric
Phi c phn ng to kt ta vi acid sulfuric c trng.
To kt ta vi ch
acetat
Phi c phn ng to kt ta vi ch acetat c trng.
5.2. tinh khit

Gim khi lng khi sy
kh
Khng c qu 20%
(Th theo hng dn trong JECFA monograph 1 - Vol.4 Sy ti 105o trong 4 gi).
Cht khng tan trong

nc
Tng propylen glycol
Khng thp hn 15% v khng c qu 45%
Propylen glycol t do
Khng c qu 15%
Arsen
Ch
5.3. Cc yu cu v vi
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QCVN 4 - 21: 2011/BYT

sinh vt
kh

my nghin cao tc.
Tng s nm men-mc
Coliforms
m tnh
Salmonella
m tnh
5.3. Hm lng CO2
Ch phm khan cha khng thp hn 16% v khng c

qu 20% CO2
6. Phng php th
6.1. nh tnh
To kt ta vi acid
sulfuric
Ly 10 ml dung dch mu th 1%, thm 1 ml dung dch natri

hydroxyd (TS). un nng trong b cch thy nc si trong 5
pht, lm ngui v thm 1 ml dung dch acid sulfuric long
(TS). Xut hin kt ta dng keo.
To kt ta vi ch
acetat
Ly 5 ml dung dch mu th 1%, thm 1 ml dung dch ch

acetat (TS). Xut hin kt ta dng keo.
6.2. tinh khit

Gim khi lng khi sy
kh
- Th theo hng dn trong JECFA monograph 1 - Vol.4

- Sy ti 105oC trong 4 gi.
Cht khng tan trong

nc

nc trong bnh 2000 ml. Trung ha ti pH 7 bng dung dch
natri hydroxyd (TS) v thm d 3 ml. Thm 40 ml dung dch
hydrogen peroxyd 30% (kl/kl), y bnh v un trong khong
1 gi, khuy thng xuyn. Lc nng qua mt chn lc
Gooch c mng lc bng x thy tinh cn b (2,4 cm, No
934 AH, Reeve Angel & Co, Clifton, N.Y., USA hoc tng
ng). Nu qu trnh lc b chm do dung dch mu c
nht cao, un si n khi nht gim n mc c th lc
c. Ra chn lc bng nc nng, sy chn lc v phn
khng tan ti 105o trong 1 gi, ngui v cn. Tnh hm
lng % ca phn khng tan sy kh.
Dch th:
Cn 1 g (chnh xc n 0,1 mg) mu th c lm kh,
ha tan trong 100 ml nc ct trong cc 400 ml. Sau khi
tan hon ton thm 50 ml dung dch natri hydroxyd 0,1 N v
khuy u trong 30 pht, cui cng trung ha bng acid
hydrocloric 0,1 N v ta gm bng 25 ml dung dch natri clorid
5%. Lc dung dch, s dng giy lc nhanh, thu dch lc vo
bnh nh mc 250 ml, ra ta vi ln bng nc ct, gp
dch ra vo dch lc v pha long n cch bng nc ct.
Tng propylen glycol
20
QCVN 4 - 21: 2011/BYT

Acid periodic 0,029 M:
Cho 5,500 g acid periodic v 200 ml nc ct vo bnh nh
mc 1000 ml. Pha long n vch bng acid acetic bng.
Tin hnh th:
Dng pipt ht 25 ml dch th v 25 ml dung dch Acid
periodic 0,029 M vo 1 bnh nn, lc u v yn trong 30
pht. Cui cng thm ~ 2 g kali iodid v chun vi dung
dch natri thiosulfat 0,1 N s dng ch th l dung dch h tinh
bt 1%. Tin hnh lm 1 mu trng song song, s dng 50 ml
nc ct thay cho dch th. Tnh hm lng tng propylen
glycol theo cng thc sau:
% propylen glycol =
Propylen glycol t do
Arsen
Ch
3,8 X (A - B)
W
Trong :
A = th tch natri thiosulfat 0,1 N chun mu trng (ml)
B = th tch natri thiosulfat 0,1 N chun mu th (ml)
W = Khi lng mu th (g)
Xc nh hm lng % ca propylen glycol t do trong mu
bng cch chit (un hi lu trong 2 gi) 2 g mu th vi 80
ml propan-2-ol. ngui v nhit phng, sau xc nh
lng propylen glycol t do bng cch chun vi Acid
periodic 0,029 M nh hng dn xc nh tng propylen
glycol.
(phng php II).
6.3. nh lng
Tin hnh theo hng dn trong chuyn lun xc nh carbon
dioxyd bng phng php decarboxyl ha ti JECFA
Mi ml natri hydroxyd 0,25N s dng tng ng vi 5,5mg
carbon dioxyd.
21
QCVN 4 - 21: 2011/BYT

Ph lc 6
YU CU K THUT V PHNG PHP TH I VI AGAR
1. Tn khc, ch s
Agar; Agar-Agar; gelose; Japan Agar; Bengal; Ceylon;

Chinese or Japanese isinglass; Layor Carang
INS 406
ADI khng gii hn
2. nh ngha
Thch l hp cht keo kh a nc chit xut t loi to

bin thuc lp Rhodophyceae. N l mt polysaccharid gm
cc tiu phn D- v L-galactose. C khong 10 tiu phn Dgalactopyranose c mt nhm ester sulfat. Cation calci,
magnesi, kali, natri cng c th lin kt vi polysaccharid
ny.
Ch s C.A.S.
9002-18-0
3. Cm quan
Khng mi hoc c mi c trng nh. Thch th khng tn

thng l hn hp nhiu dng mng mng, dnh; hoc bt,
ht dng vy, ming. C th c mu cam vng nht hoc
vng xm n vng nht hoc khng mu. Dai khi ht m v
rn khi kh. Bt thch c mu trng, trng vng, vng nht.
4. Chc nng
Cht lm dy, cht n nh, cht nh ha.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc lnh, tan trong nc nng.
To gel vi nc
Phi c phn ng to gel vi nc c trng.
To kt ta vi dung
dch amoni sulfat
Phi c phn ng to kt ta vi dung dch amoni sulfat c

trng.
To kt ta vi dung
dch ch acetat
Phi c phn ng to kt ta vi dung dch ch acetat c

trng.
Kim tra hin vi
t mt vi mnh thch cha nghin hoc mt t bt thch

trn lam knh, thm vi git nc hoc dung dch cloral
hydrat (TS). Soi di knh hin vi, thch trong nc c dng
ht xen ln dng si., thch trong dung dch cloral hydrat
(TS) trong hn thch trong nc.
5.2. tinh khit

Hp th nc
t yu cu (m t trong phn Phng php th).
Gim khi lng khi sy

kh
Khng c qu 22%.
Tro ton phn
Khng c qu 6,5% tnh theo ch phm lm kh.
Tro khng tan trong acid Khng c qu 0,5% tnh theo ch phm lm kh.
22
QCVN 4 - 21: 2011/BYT

Cc tp cht ngoi lai
khng tan trong nc
Khng c qu 1,0%.
Tinh bt v dextrin
Khng pht hin c.
Gelatin v cc protein
khc
Khng pht hin c.
Arsen
Ch
5.3. Cc yu cu v vi
sinh vt
kh

my nghin cao tc.
Tng s nm men-mc
Coliforms
m tnh
Salmonella
m tnh
5.4. Hm lng
Ngng nng gel khng c qu 0,25%.
6. Phng php th
6.1. nh tnh
To gel vi nc
Pha dung dch mu th 1% trong nc si, cho vo bnh,

t bnh trong b cch thy 30o trong 15 pht. To thnh gel
rn v bn. t bnh trong b cch thy 70o trong 1 gi. Gel
khng b chy. Khi gia nhit n > 95o, gel b chy to thnh
dung dch trong.
To kt ta vi dung
dch amoni sulfat
Dung dch mu th 0,5% (m khong 40o) to kt ta khi

thm dung dch amoni sulfat (m khong 40o) vi th tch
bng 1/2 th tch dung dch mu th. Php th ny phn bit
thch vi cc alginat, gm arabic, gm ghatti, gm karaya,
pectin v tragacanth.
To kt ta vi dung
dch ch acetat
Dung dch mu th 0,5% (m) to kt ta khi thm dung dch

ch acetat (m) vi th tch bng 1/5 th tch dung dch mu
th. Php th ny phn bit thch vi methyl cellulose.
6.2. tinh khit

Hp th nc
Cho 5 g mu th vo trong mt ng ong 100 ml, cho nc

n vch, khuy u, yn ti 25o trong 24 gi. Rt dung
dch qua bng thy tinh thm nc, thu nc chy ra vo
mt ng ong khc. Lng nc chy ra thu c khng
c qu 75 ml.
23
QCVN 4 - 21: 2011/BYT

Gim khi lng khi lm - Th theo hng dn ti JECFA monograph 1 - Vol.4.
- Sau khi sy ti 105o n khi s chnh lch khi lng gia
kh
2 ln cn khng qu 1 mg (thi gian sy khong 5 gi);
thch th cha nghin cn c ct nh thnh tng ming 2
-5 mm2 trc khi sy).
Cc tp cht ngoi lai
khng tan trong nc
un si, hi lu 5 g mu th vi 500 ml nc v 12 ml acid

sulfuric. ngui v lc qua mt phu thy tinh xp, l mn,
cn b. Trng bnh v lc bng 50 ml nc, sy ti 105o
n khi lng khng i v cn. Tnh hm lng %.
Tinh bt v dextrin
Thm vo dung dch mu th 0,5% (m khong 40o) 2 git

dung dch iod (TS). Khi va nh thuc th vo dung dch th
c mu tm nhng sau khi khuy trn u dung dch c
mu nu vng khng phi mu xanh hoc mu .
Gelatin v cc protein
khc
Thm vo dung dch mu th 0,5% (m khong 40o) dung

dch acid picric (TS) (m khong 40o). Sau 10 pht dung dch
khng c c.
Arsen

(phng php II).
Ch

6.3. nh lng
Ngng nng gel:
Pha mt lot dung dch mu th cha hm lng mu th
rn 0,15%; 0,20%; 0,25%..., cho vo cc ng nghim kch
thc di 150 mm, ng knh trong 16 mm. Nt ng
nghim v lm mt trong 1 gi ti 20 - 25o. ct gel t cc
ng nghim ln trn 1 b mt phng. Nng thp nht chu
c trng lc trong 5 - 30 giy m khng b gy v l
ngng nng gel ca mu th.
24
QCVN 4 - 21: 2011/BYT

Ph lc 7
YU CU K THUT V PHNG PHP TH I VI CARRAGEENAN
1. Tn khc, ch s
Thch trng t rong bin Ailen ( t loi Chondrus spp.);

Eucheuman (t loi Eucheuma spp. ); Iridophycan (t loi
Iridaea spp.); Hypnean (t loi Hypnea spp.); Furcellaran
hoc agar an Mch (t loi Furcellaria fastigiata);
INS 407
2. nh ngha
Sn phm c tnh keo a nc thu c t mt s loi

thuc lp to bin Rhodophyceae.
Carrageenan c ly ch yu t cc h v loi ca lp to
bin Rhodophyceae gm:
Furcellariacaea nh Furcellaria
Gigartinaceae nh Chondrus, Gigartina, Iridaea
Hypnaeceae nh Hypnea
Phyllophoraceae
nh
Phyllophora,
Gynmogongrus,
Ahnfeltia
Solieriaceae nh Eucheuma, Anatheca, Meristotheca.
Carrageenan l cht keo a nc gm cc thnh phn
chnh l cc ester ca ng galactose v 3,6anhydrogalactose polysaccharide vi (NH4)2SO4, CaSO4,
MgSO4, K2SO4 v Na2SO4. Cc ng hexoza ny c lin
kt -1,3 v -1,4 trong copolymer. T l tng i gia cc
cation tn ti trong carrageenan c th thay i trong qu
trnh ch bin khi mt cation chim phn chnh.
Cc polysaccarid ph bin trong carrageenan l kappa-,
iota- v lambda-carrageenan. Kappa-carrageenan thng
dng polymer ca D-galactose-4-sulfat v 3,6- anhydro-Dgalactose; iota-carrageenan cng tng t nh vy, ngoi
tr 3,6-anhydrogalactose c sulfat ho v tr cacbon s
2. Gia kappa-carrageenan v iota-carrageen c mt hp
phn trung gian khc nhau v mc sulfat ho v tr
cacbon s 2. i vi lambda-carrageenan, cc n v ng
phn ch yu l D-galactoza-2-sulfat (lin kt 1,3) v Dgalactoza-2,6-disulfat (lin kt 1,4).
Carrageenan thu c t qu trnh chit to bin bng
nc hoc dung dch kim long. Thu hi carrageenan
trong dch chit bng phng php kt ta cn hoc
phng php kt ta vi dung dch kali clorid v lm lnh
ng. Ch s dng methanol, ethanol v isopropanol thu
hi v tinh ch. Ngoi ra, ch phm thng mi c th c
th cha ng ( chun ho), mui ( lm dy v keo
ho).
M s C.A.S.
9000-07-1
3. Cm quan
Dng bt th ti mn, mu hi vng hoc mu nu vng ti

trng, hu nh khng mi
25
QCVN 4 - 21: 2011/BYT

4. Chc nng
Cht lm dy, cht to gel, cht n nh, cht nh ho
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong ethanol ; tan trong nc nhit khong

800C, to dung dch nht trong hoc hi trng sa chy d
dng ; tan trong nc d dng hn nu trc c lm
m bng cn, glycerol hoc dung dch bo ho ca glucose
hoc sucrose trong nc.
Sulfat
Phi c phn ng c trng ca sulfat.
Galactose v
anhydrogalactose
Phi c phn ng
anhydrogalactose.
trng
ca
galactose
Keo a nc v cht Phi c phn ng c trng ca keo a nc v cht ng

ng trng hp in trng hp in hnh.
hnh
Hp th tia hng ngoi
Phi c phn ng hp th tia hng ngoi c trng.
5.2. tinh khit

0
Gim khi lng khi sy Khng c qu 12% (nhit sy 105 C n trng lng
khng i).
kh
pH
8 11 (th vn 1%)
nht
Khng c nh hn 5 cP 750C (dung dch 1,5%)
Sulfat
Khng c nh hn 15,0% v khng c qu 40,0%

(SO42-) tnh theo trng lng kh.
Hm lng tro ton

phn
Khng c nh hn 15,0% v khng c qu 40,0%

theo trng lng kh
Tro khng tan trong

acid
Khng c qu 1,0%
Cht khng tan trong

acid
Khng c qu 2,0%
S dng 2 g thu c t phn (a) ca Cch tin hnh xc
nh sulfat
D lng dung mi
Khng c qu 0,1% ethanol, isopropanol, hoc methanol,

dng mi cht hay cng hp cc cht.
Xc nh theo m t trong phn Phng php th - nh
lng
26
QCVN 4 - 21: 2011/BYT

Arsen
Khng c qu 3,0 mg/kg (Xc nh bng k thut quang

ph hp th nguyn t H+ i vi 3 g mu).
Ch
Cadmi
Thu ngn
5.3. Cc yu cu v vi
sinh vt
kh
Khng c qu 5.000 cfu/g
Salmonella spp.
m tnh
E.coli
m tnh/1 g
6. Phng php th
6.1. nh tnh
Sulfat
Ho tan 100 mg mu trong 20 ml nc (gia nhit nu cn),

thm 3 ml dung dch BaCl2 TS v 5 ml dung dch acid HCl
long; lc nu c kt ta. un si dung dch hoc phn dch
c lc ra trong 5 pht. Xut hin kt ta tinh th trng.
Galactose v
anhydrogalactose
Tin hnh theo ch dn di phn nh tnh cc thnh phn

gm (Quyn 4) s dng cc cht chun sau y lm i
chiu: galactose, rhamnose, acid galacturonic, 3,6
anhydrogalactose, mannose, xylose v arabinose.
Galactose v 3,6 anhydrogalactose cn phi hin din.
Keo a nc v cht Cho 4 g mu vo 200 ml nc v gia nhit hn hp trong

0
ng trng hp in chu nc 80 C, khuy vi tc nht nh cho n khi
ho tan. B sung lng nc mt i do qu trnh bay hi v
hnh
lm ngui dung dch n nhit phng. Dung dch s tr
nn nht v to gel. Thm 200 mg KCl vo 50 ml dung dch
hoc gel, sau li gia nhit, khuy u v lm ngui.
Gel gin, d v ch ra carrageenan chim u th dng
kappa ; v gel mm do, n hi ch ra carrageenan chim
u th dng iota. Nu dung dch khng to gel,
carrageenan chim u th dng lambda.
27
QCVN 4 - 21: 2011/BYT

Hp th tia hng ngoi
Ghi ph hp th tia hng ngoi trn cc phn gel v phn

khng gel ca mu theo cch sau:
Ho u 2 g mu trong 200 ml dung dch KCl 2,5%, khuy
trong 1 gi. qua m, khuy li trong 1 gi v chuyn
vo mt ng li tm (nu khng chuyn c v h phn tn
qu nht, pha long vi 200 ml dung dch KCl). Li tm
khong 1.000 xg trong 15 pht .
B phn trong trn b mt, ho tan phn cn thu c trong
200 ml dung dch KCl 2,5%, li tm li. Lm ng t phn ni
trn b mt bng cch thm gp 2 ln th tch ethanol 85%
hoc isopropanol (Lu : gi li phn lng xung s dng
theo cch lm sau). Thu hi phn ng t v ra bng 250
ml cn. p dch lng ca phn ng t ra ngoi v sy kh
600C trong 2 gi. Sn phm thu c l phn khng gel
(lambda-carrageenan).
Ho u phn lng xung ( c gi li trn) trong 250
ml nc lnh, gia nhit 900C trong 10 pht, lm ngui ti
600C. Lm ng t hn hp v sau thu hi, ra v sy
phn ng t theo cch lm trn. Sn phm thu c l
phn gel (kappa v iota-carrageenan).
Chun b mt dung dch nc 0,2% cho mi phn, soi phim
c dy 0,5 mm (khi sy) trn b mt khng dnh nh
Teflon v thu c ph hp th tia hng ngoi ca mi phim
(Ngoi ra, ph hp th c th thu c nh k thut soi
phim trn tm knh ph kali bromid nu vic thc hin trnh
c m).
Carrageenan c ph hp th rng v mnh, c trng ca
tt c polysaccharid trong vng t 1.000 n 1.100 cm-1.
Hp th ti a l 1.065 v 1.020 cm-1 i vi dng gel v
dng khng gel. Cc di hp th c trng khc v cng
hp th tng ng vi hp th ti 1.050 cm-1 nh sau:
S
sng
(cm-1)
Phn t
hp th tng ng vi
1.050 cm-1
Kappa
Iota
Lam
bda
1.2201.260
Ester sulfat
0,2-1,2
1,2-1,6
1,42,0
928933
3,6-anhydro
galactose
0,2-0,6
0,2-0,4
0-0,2
840850
Galactose-4sulfat
0,1-0,5
0,2-0,4
825830
Galactose-2sulfat
0,20,4
810820
Galactose-6sulfat
0,10,3
28
QCVN 4 - 21: 2011/BYT

800805
3,6-anhydro
galactose-2sulfat
0-0,2
0,2-0,4
29
QCVN 4 - 21: 2011/BYT

6.2. tinh khit
Sulfat
Nguyn tc: Cc nhm sulfat thu phn c kt ta di

dng BaSO4
Cch tin hnh:
(a) Ho tan chnh xc 8 g mu sn phm thng mi trong
400 ml hn hp isopropanol/nc 60% (theo khi lng)
nhit phng. Khuy nh nhng trong 4 gi. Lc bng
giy lc khng tro. Loi b phn dch lc c. Ra phn
cn li trn giy lc 2 ln bng 10 ml hn hp
isopropanol/nc 60%. Sy n trng lng khng i
1050C.
Phn (b) dng khong 1 g cht kh. Phn cn li c gi
li xc nh hm lng tro ton phn v hm lng cht
khng tan trong dung dch acid.
(b) Cn chnh xc 1 g mu (W 1) thu c phn (a) cho
vo bnh y trn c di dung tch 100 ml. Thm 50 ml dung
dch acid HCl. Ni bnh trn vi mt sinh hn c t nht 5
bu ngng v un si vi sinh hn ngc trong 1 gi. Thm
25 ml dung dch hydrogen peroxyd v tip tc un si vi
sinh hn ngc trong khong 5 gi hoc cho n khi dung
dch trong sut hon ton. Chuyn dung dch vo cc thu
tinh dung tch 600 ml, un si v thm 10 ml dung dch
BaCl2 theo tng git. Gia nhit hn hp phn ng trong 2
gi bng cch thy si. Lc hn hp qua giy lc khng tro.
Ra bng nc ct un si cho n khi dch lc khng cn
clorid. Sy giy lc v cc cht trong t sy. t 8000C
trong chn s hoc chn silica cho n khi tro chuyn thnh
mu trng. ngui trong bnh ht m.
Cn chn c tro. Tnh % sulfat t khi lng gam (W 2) ca
tro (BaSO4) s dng cng thc: (W 2/W 1) x 100 x 0,4116
Hm lng tro ton

phn
Cho chnh xc 2 g mu kh (W 1) thu c t phn (a) trong

cch tin hnh xc nh sulfat trn vo chn silica hoc
ni platin. Gia nhit mu bng n hng ngoi thch hp,
tng cng dn dn cho n khi mu hon ton b than
ho; tip tc gia nhit thm khong 30 pht. Chuyn chn
trn c cha mu than ho vo l kn v nung 5500C
trong 1 gi. Lm ngui trong bnh ht m v cn. Lm li
vic nung trong l kn cho n khi t c khi lng
khng i (W 2). Nu tro khng cn carbon khng thu c
sau ln nung th nht, lm m phn c than ho bng
dung dch NH4NO3 v sy bng n hng ngoi. Lm li
bc nung.
Tnh % tro ton phn trong mu: (W 2/W 1) x 100
Gi li tro cho nh lng tro khng tan trong acid.
30
QCVN 4 - 21: 2011/BYT

nht
Cho 7,5 g mu v 450 ml nc kh ion vo cc

Berzelius dung tch 600 ml, khuy t 10 20 pht. Thm
vo lng nc cn thit t khi lng cui cng 500
g, khuy v gia nhit trong cch thy cho n khi nhit
t 800C (khong 20 30 pht). Cho nc thm vo b
vo lng nc mt i do bc hi, lm ngui ti 76 770C,
gia nhit trong cch thy nhit khng i 750C. Gia
nhit trc phao v b phn bo v ca nht k Brookfield
LVF hoc LVT khong 750C trong nc. Sy phao v b
phn bo v v lp chng vo nht k c trang b trc
quay s 1 (ng knh 19 mm, chiu di khong 65 mm) c
kh nng quay 30 vng/pht. iu chnh cao ca phao
trong dung dch mu, bt u quay nht k 30 vng/pht v
sau su vng quay ca nht k, ly nht k c mc 0
100.
Nu nht rt thp, chnh xc tng ln c th thu c
bng cch s dng UL Brookfield (cc thp) b tip hp
hoc tng ng. (Lu : mu ca mt vi dng
carrageenan c th qu nht c khi s dng trc quay
s 1). Nhng mu nh vy r rng l vt qua cc c
im k thut, nhng nu c nht mong mun v cc l
do khc, s dng trc quay s 2 v c mc 0 - 100 hoc
mc 0 - 500).
Ghi li cc kt qu (n v cP) thu c bng cch c trn
nht k theo h s c a ra bi nh sn xut
Brookfield.
31
QCVN 4 - 21: 2011/BYT

D lng dung mi
Dung dch alcol chun:

Cho 500 mg mi loi methanol, ethanol v isopropanol c
cht lng dng cho sc k vo bnh nh mc dung tch 50
ml, pha long bng nc cho th tch 50 ml, lc u.
Dng pipet ly 10 ml ca dung dch trn vo bnh nh mc
100 ml, pha long bng nc cho th tch 100 ml, lc
u.
Dung dch TBA chun:
Cho 500 mg tertiary-butyl alcol c cht lng dng cho sc
k vo bnh nh mc dung tch 50 ml, pha long bng nc
cho th tch 50 ml, lc u. Dng pipet ly 10 ml ca
dung dch trn vo bnh nh mc 100 ml, pha long bng
nc cho th tch 100 ml, lc u.
Hn hp dung dch chun:
Dng pipet ly 4 ml mi loi dung dch alcol chun v dung
dch TBA chun cho vo bnh Erlenmeyer dung tch 125 ml,
pha long n th tch khong 100 ml bng nc v lc
u. Dung dch ny cha khong 40 g mi alcol/ml.
Chun b dung dch mu th:
Ho tan 1 ml nh tng chng bt thch hp nh DowCorning G-10 hoc tng ng trong 200 ml nc ng
trong bnh chng ct y trn 24/40 dung tch 1.000 ml.
Thm vo 5 g mu v lc bng my lc trong 1 gi. Ni bnh
chng ct vi ct phn on v chng ct khong 100 ml,
iu chnh nhit bt khng c vo trong ct. Thm 4,0
ml dung dch chun TBA vo phn chng ct c c
dung dch mu th.
Cch tin hnh:
Bm 5 l hn hp dung dch chun vo sc k kh thch hp
c trang b detector ion ho ngn la v ct thp khng r
1,8-m x 3,2-mm c nhi bng Porapak QS 80/100-mesh
hoc tng ng. Cht mang l kh heli vi tc dng
80 ml/pht. Nhit bung bm mu 2000C, nhit ct
1650C v nhit detector 2000C. Thi gian lu ca
isopropanol khong 2 pht v thi gian lu ca tertiary-butyl
alcol khong 3 pht.
Tnh din tch pic ca methanol, ethanol, isopropanol v
TBA. Tnh h s p ng ca mi loi fi theo cng thc
Ai/ATBA, trong Ai l din tch pic ca mi alcol (I =
methanol, ethanol hoc isopropanol).
Tng t, bm 5 l dung dch mu th v tnh cc din tch
pic, ghi din tch ca mi pic alcol l Ai v din tch pic ca
tertiary-butyl alcol l ATBA.
Tnh hm lng tng alcol (mg/kg) c trong mu theo cng
thc:
Ai 4000 / fi ATBA W
Trong : W l khi lng mu ly (g)
32
QCVN 4 - 21: 2011/BYT

Ch
Nguyn tc:
Mu c tro ho, lm m bng cc acid nitric, acid
perchloric v c phn tch bng cch o ph hp th
nguyn t ngn la (Quyn 4).
Thit b:
My o ph hp th nguyn t
Ho cht:
- Acid nitric, m c, tinh khit
- Acid perchloric, m c, tinh khit
- Acid hydrochloric, m c, tinh khit
- Dung dch chun ch ( cng b)
Dung dch:
- Dung dch gc (1mg/ml): Pha mt th tch thch hp ca
dung dch chun ch cng b vi nc ct v kh ion
(D/D nc) c 1 lt dung dch.
- Dung dch trung gian: (a) 100 g/ml: Dng pipet ly 10 ml
dung dch gc cho vo bnh nh mc dung tch 100 ml v
pha n th tch 100 ml bng D/D nc. (b) 10 g/ml. Dng
pipet ly 10 ml ca dung dch 100 g/ml cho vo bnh nh
mc dung tch 100 ml v pha n th tch 100 ml bng D/D
nc.
- Dung dch chun: Ly 4 bnh nh mc dung tch 100 ml v
chuyn vo chng ln lt 1, 5, 10, 20 ml dung dch ch
trung gian (b). Pha n th tch 100 ml bng D/D nc
to cc dung dch c nng l 0,1; 0,5; 1 v 2 g Pb/ml.
Chun b mu: (Ch : cch lm ny s dng cc acid oxy
ho m c v dn n bay hi ca cc kh c. Phi thc
hin trong t ht).
Cn chnh xc 7,5 g mu th dng bt kh i din cho vo
bnh Erlenmeyer dung tch 250 ml. Lm mu trng v thc
hin cc bc tng t nh vi mu th. Lm m mu th
bng khong 10 ml D/D nc v thm 25 ml dung dch acid
nitric. Gia nhit nh nhng trn tm gia nhit (100 1500C)
cho n khi hu ht khi en thot ra (khong 1 gi); Thnh
thong lc bnh. Lm ngui v thm 5 ml dung dch acid
perchloric; giai on ny cc cu t tr nn d nhn thy.
Gia nhit nh nhng (bng tm gia nhit, 100 1500C)
c cho n khi dung dch chuyn sang mu hi vng hoc
khng mu (khong 1 gi). gia qu trnh gia nhit nu
dung dch sm mu, thm t t my ln 2 3 ml dung dch
acid nitric nu cn thit cho n khi t c mu mong
mun; khng c dung dch cn. Lm ngui phn
gia nhit v ra thnh bnh bng khong 5 ml D/D nc v
lc xoy. B sung 2 ml dung dch acid hydrochloric. Gia
nhit li cho n khi tt c khi mu nu c thot ra v
dung dch c mu trng ti vng nht. Khng c dung
dch cn. Lm ngui dung dch v ra thnh bnh bng
khong 10 ml D/D nc. Chuyn dung dch hi nht vo
bnh nh mc dung tch 50 ml v pha n th tch 50 ml
33
QCVN 4 - 21: 2011/BYT

bng D/D nc. Lc bng giy lc 2 lp (Whatman s 5
hoc loi tng ng).
nh lng:
Cho vo my o ph hp th nguyn t trc thit
lp cc iu kin ti u bc sng 283,3 nm s dng
khng kh/acetylen oxy ho ngn la. o hp th ca
mu th, mu trng v dung dch chun. V ng chun
trn th hp th vi nng ch (g Pb/ml) ca mu
trng v dung dch chun. nh lng nng ch trong
dung dch mu th t ng chun.
Nng ch trong mu th (mg Pb/kg):
[Pb] = F x A/B
Trong : A l nng ch trong dung dch mu (g/ml)
B l khi lng mu th (g)
F l h s pha long (50 ml)
34
QCVN 4 - 21: 2011/BYT

Cadmi
Thc hin nh hng dn trn i vi nh lng ch, s

dng bc sng phn tch 228,8 nm. Dung dch trung gian
v dung dch mu c chun b t dung dch chun cadmi
cng b:
- Dung dch trung gian: (a) 100 g/ml: Dng pipet ly 10 ml
dung dch gc (1 mg/ml) cho vo bnh nh mc dung tch
100 ml v pha n th tch 100 ml bng D/D nc. (b) 10
g/ml: Dng pipet ly 10 ml ca dung dch (a) cho vo bnh
nh mc dung tch 100 ml v pha n th tch 100 ml bng
D/D nc. (c) 1 g/ml: Dng pipet ly 1 ml dung dch (a) cho
vo bnh nh mc 100 ml, pha n th tch 100 ml bng
D/D nc.
chuyn vo chng ln lt 0,5; 2,5; 5, 10, 20 ml dung dch
trung gian (c). Pha n th tch 50 ml bng D/D nc to
cc dung dch c nng l 0,01; 0,05; 0,1; 0,2 v 0,4 g
Cd/ml.
Nng cadmi trong mu th (mg Cd/kg):
[Cd] = F x A/B
Trong : A l nng cadmi trong dung dch mu (g/ml)
Thu ngn
Nguyn tc:
Mu c tro ho, lm m bng cc acid nitric, acid
perchloric v c phn tch bng cch o ph hp th
nguyn t H+ (Quyn 4)
Thit b:
My o ph hp th nguyn t c trang b vi thit b to
hi hydrogen. Khng th thiu ng phn ng hoc cun dy
v bm pittng vi cc knh ng kp: mt knh dnh cho
dung dch mu v mt knh dnh cho 2 ng dung dch ho
cht. Kim sot dng chy c xc nh bi c ng v u
ni ng. Tc dng c o ng ra ca thit b to
kh hydrogen.
35
QCVN 4 - 21: 2011/BYT

Ho cht:
- Acid nitric, m c, tinh khit
- Acid perchloric, m c, tinh khit
- Acid hydrochloric, m c, tinh khit
- Natri borohydrid, > 98%
- Natri hydroxyd, tinh khit
- Dung dch chun thu ngn ( cng b)
Dung dch:
- Hn hp dung dch acid nitric v acid perchloric (1 : 1):
Trn theo t l th tch nh nhau ca hai acid.
- Dung dch acid HCl 5M: Pha 417 ml dung dch acid HCl
m c n th tch 1 lt bng nc kh ion.
- Dung dch natri borohydrid 0,4% (chun b ngay trc khi
dng): Trc tin, ho tan 2,5 g NaOH trong nc kh
ion. Sau , thm v ho tan 2,0 g natri borohydrid. Pha ti
th tch 500 ml.
- Dung dch gc (1 mg/ml): Pha mt th tch thch hp dung
dch chun thu ngn cng b bng nc ct v kh
ion (D/D nc) ti th tch 1 lt.
- Dung dch trung gian: (a) 10.000 g/l: Dng pipet ly 1 ml
dung dch gc cho vo bnh nh mc dung tch 100 ml v
pha n th tch 100 ml bng D/D nc. (b) 100 g/l: Dng
pipet ly 1 ml dung dch 10.000 g/l cho vo bnh nh mc
dung tch 100 ml v pha n th tch 100 ml bng D/D
nc.
chuyn vo chng ln lt 1; 5, 10, 15 v 20 ml dung dch
trung gian (b). Mi bnh b sung 10 ml hn hp dung dch
acid nitric v acid perchloric (1:1). Pha n th tch 100 ml
bng D/D nc to cc dung dch c nng l 1; 5; 10;
15 v 20 g Hg/l.
Chun b mu: (Ch : cch lm ny s dng cc acid oxy
ho m c v dn n bay hi ca cc kh c. Phi thc
hin trong t ht).
Cn chnh xc 5 g mu th dng bt kh i din cho vo
bnh Erlenmeyer dung tch 250 ml. Lm mu trng v thc
hin cc bc tng t nh vi mu th. Lm m mu th
bng 5 ml D/D nc v thm 10 ml hn hp dung dch acid
nitric v acid perchloric (1:1). Gia nhit nh nhng trn tm
gia nhit (100 1500C) cho n khi hu ht khi en thot
ra v dung dch chuyn sang mu vng nht hoc khng
mu; Thnh thong lc bnh. Khng c dung dch cn.
Lm ngui v ra thnh bnh bng lng nh D/D nc.
(Mt vi cu t c th nhn thy c). y bnh nh
nhng. dung dch nht khng ng k qua m. Chuyn
dung dch vo bnh nh mc dung tch 50 ml v pha n
th tch 50 ml bng D/D nc. Lc vo bnh Erlenmeyer
bng giy lc 2 lp (Whatman s 5 hoc loi tng ng).
36
QCVN 4 - 21: 2011/BYT

Nhng bnh vo chu siu m v bt ch rung siu m
trong 10 pht hoc cho n khi bt khng c to thnh
trn b mt dung dch.
nh lng: Hiu chnh (s dng nc) my bm pittng
a ra tc dng ca dung dch mu 8 ml/pht v tc
dng kt hp ca hai dung dch (Natri borohydrid v acid
hydrochloric 5M) 2 ml/pht. (Tc dng kt hp c thc
hin vi mt thit lp bm duy nht).
t mu vo my quang ph trc thit lp cc iu
kin ti u bc sng n i vi thu ngn 253,7 nm.
Chuyn lng thch hp ca hai dung dch ho cht vo cc
ng ong c chia vch nh mc ring bit. t ng ht
ring bit t bm pittng vo mi dung dch ho cht v
bnh cha mu. Bt u dng ca kh mang argon (p sut
u ra ca thng cha: 3,2 0,2 kg/cm2) thng qua thit b
to hi hydrogen ca my quang ph. Khi ng bm
bt u dng ca ba dung dch trong ng gp ca thit b
to hydrogen. Ti ng gp cc dung dch c trn u vi
nhau v chy trong ng xon rut g to ra thu ngn
nguyn t v thu ngn nguyn t c mang n bung
hp th ca my quang ph hp th. o hp th cho
mu. Lp li i vi dung dch mu trng v mi dung dch
chun.
V ng chun trn th hp th vi nng thu
ngn (g Hg/l) ca mu trng v dung dch chun. nh
lng nng thu ngn trong dung dch mu th t
ng chun.
Nng thu ngn trong mu th (mg Hg/kg):
[Hg] = F x A/1000B
Trong : A l nng thu ngn trong dung dch mu (g/l)
5.3. Cc yu cu v vi
sinh vt
Thm 50 g mu vo 450 ml dung dch pha long phosphat

m ca Butterfield v ng ho hn hp trong mt my
trn tc cao.
37
QCVN 4 - 21: 2011/BYT
38
QCVN 4 - 21: 2011/BYT

Ph lc 8
YU CU K THUT V PHNG PHP TH I VI GM U CAROB
1. Tn khc, ch s
Locust bean gm, carob gm

INS 410
2. nh ngha
Xut x l ni nh c nghin ca cc ht t Ceratonia

siliqua (L) Taub. (Fam. leguminosae) ch yu bao gm khi
lng phn t cao (khong 50.000 3.000.000) cc
polysaccharid gm c cc galactomannan. Cc ht c tch
v ngoi bng cch x l nhn ht vi acid sulfuric long
nhit cao hoc rang nhn ht, tip theo l xay v sng ht
c c nhng ni nh. Gm c lm sch bng cch
ra vi ethanol hoc isopropanol hoc ho trong nc si,
sau lc, lm bc hi dung mi v sy kh.
M s C.A.S.
9000-40-2
3. Cm quan
Dng bt mu trng n mu trng hi vng, gn nh khng

c mi
4. Chc nng
Cht lm dy, cht n nh, cht nh ho
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong ethanol
To gel
Phi c phn ng to gel c trng.
nht
Phi c php th c trng ca nht.
Thnh phn gm
Phi c phn ng c trng ca thnh phn gm.
Kim tra hin vi
Phn tn mu gm trong dung dch nc cha 0,5% iod v

1% kali iodid tri ln tiu bn knh v soi bng knh hin vi.
Gm u carob cha cc t bo dng ng cng di, tch bit
hoc hi an xen nhau. Cc thnh phn ca gm u carob
c mu nu c hnh thnh t thng xuyn hn gm
Guar. (Gm Guar cho thy cc nhm t bo dng hnh trn
ti dng qu l. Cc thnh phn ca gm Guar c mu vng
ti mu nu).
5.2. tinh khit

Gim khi lng khi
sy kh
Khng c qu 14,0% (nhit sy 1050C trong 5 gi).
Tro tng s
Khng c qu 1,2% (nhit sy 8000C trong 3 4 gi).
Cht khng tan trong

acid
Khng c qu 4,0% (Th vi chnh xc 1,5 g mu th)
Protein
Khng c qu 7,0%.
39
QCVN 4 - 21: 2011/BYT

Tinh bt
Khng pht hin c.
Ethanol v
isopropanol
Khng c qa 1%, dng n cht hoc hp cht

(m t phn Phng php th)
Ch
5.3 Cc yu cu v vi
sinh vt
Tng s vi sinh vt
Khng c qu 5.000 CFU/g
E.coli
m tnh i vi mu th
Salmonella
m tnh i vi mu th
Nm men v nm
mc
Khng c qu 500 CFU/g
6. Phng php th
6.1. nh tnh
To gel
Thm lng nh dung dch natri borat TS vo dung dch mu

c phn tn bng nc; gel c hnh thnh.
nht
Cho 2 g mu vo trong cc dung tch 400 ml v lm m hon

ton bng 4 ml isopropanol. B sung 200 ml nc v khuy
mnh, tip tc khuy cho n khi gm c phn tn u
hon ton. Mt dung dch hi nht mu trng sa c hnh
thnh. Chuyn 100 ml dung dch ny vo mt cc khc dung
tch 400 ml. Gia nhit hn hp trong cch thy si trong 10
pht v lm ngui ti nhit phng. C s tng nht
ng k (y l s khc bit gia gm u carob vi gm
Guar).
Thnh phn gm
Tin hnh nh hng dn phn nh tnh thnh gm (Vol.

4) s dng 100 mg mu thay th cho 200 mg v 1 10 l
hydrolysat thay th cho 1 5 l. S dng galactose v
mannose nh l cht chun i chng. Cc thnh phn ny
lun c mt.
6.2. tinh khit

Protein
Tin hnh nh hng dn phn nh lng nitrogen

(Phng php Kjeldahl). % protein trong mu bng %
nitrogen nh lng c nhn vi 6,25.
Tinh bt
Thm mt vi git iod TS vo dung dch mu 1/10. Khng

c xut hin mu xanh.
40
QCVN 4 - 21: 2011/BYT

Ethanol v isopropanol
Nguyn tc:
Cc alcol c chuyn thnh cc ester nitrit tng ng v
c xc nh bng sc k kh khng gian hi.
Chun b mu:
Ho tan 100 mg mu trong 10 ml nc s dng NaCl nh l
cht lm phn tn nu cn.
Dung dch ni chun:
Chun b dung dch nc cha 50 mg/l n propanol
Chun b dung dch nc cha 50 mg/l mi loi ethanol v
isopropanol
Cch tin hnh:
Cn 200 mg ure cho vo mt l sm mu dung tch 25 ml
(Reacti-flasks, Pierce, Rockford, IL, USA, hoc tng
ng). Lm trong bng nitrogen trong 5 pht v sau thm
1 ml dung dch acid oxalic bo ho, ng bng nt cao su v
lc xoy. Thm 1 ml dung dch mu, 1 ml dung dch ni chun
v ng thi bt u bm gi (T = 0). Lc xoy l v y nt
li bng nt xoy c m cao su silicon. Lc xoy cho n khi
T = 30 giy. Ti thi im T = 45 giy, bm qua m cao su
silicon 0,5 ml dung dch natri nitrit (250 g/l). Lc mnh cho n
khi T = 70 giy v T = 150 giy ht qua m cao su silicon 1
ml khng gian hi s dng syringe kho p (Precision
Sampling Corp., Baton Rouge, Louisiana, USA, hoc tng
ng).
Sc k kh:
a kim syringe vo cng bm; y mu sau m syringe
v bm mu.
S dng cc iu kin sau:
- Ct: thu tinh (ng knh trong 4 mm, chiu di 90 cm)
- Cht nhi: 15 cm u c nhi bng chrompack (hoc
tng ng) v phn cn li c nhi bng Porapak R
120 -150 mesh (hoc tng ng)
- Kh mang: nitrogen (tc dng: 80 ml/pht)
- Detector: ion ho ngn la
- Nhit : cng bm 2500C, ct 1500C ng nhit
Tnh kt qu:
nh lng ethanol v isopropanol c mt trong mu bng
cch so snh cc din tch pic vi cc pic thu c nh sc
k kh khng gian hi c hnh thnh do thay th 1 ml dung
dch chun alcol cho 1 ml dung dch mu trong Cch tin
hnh nu trn.
41
QCVN 4 - 21: 2011/BYT

Ch

6.3. Ch tiu vi sinh vt

Tng s vi sinh vt
Dng k thut v trng, pha 1 g mu trong 99 ml dung dch

m phosphat v dng Stomacher, my lc hoc my
khuy ho tan hon ton. Gii hn thi gian ho tan
khong 10 pht v sau dng pipet ly 1 ml dung dch vo
cc a petri ring bit, cp i v nh du thch hp.
vo mi a petri khong 12 15 ml agar PCA trc
c lm ngui n 44 460C. Trn u bng cch quay
o lt di chuyn trc, sau cc a, cho agar ng
c. Lt ngc cc a petri v trong 48 2 gi 35 10C.
Sau khi , m cc khun lc pht trin nhn thy c trn
mi a v ghi li s khun lc. Ly trung bnh ca hai a
v nhn vi h s pha long mu 100. Trng hp khng
c khun lc c nhn thy, kt qu c th hin l nh
hn 100 CFU/g.
E.coli
S dng cellulase phn gii mu gm trc khi phn

tch l cn thit trnh to gel ca gm trong sut qu
trnh thm gm vo canh thang tng sinh. Chun b dung
dch cellulase 1% (1 g cellulase trong 99 ml nc) v tit
trng bng cch lc qua mng 0,45 m (Dung dch cellulase
c th bo qun 2 50C trong 2 tun). Cho vo ng tit
trng c cha 9 ml canh trng c lauryl sulfat tryptose
(LST), thm vo 0,1 ml dung dch cellulase 1% tit trng.
B sung 1 g mu gm vo ng v lc mnh bng my lc
Vortex phn tn mu. ng trong 24 48 gi 35 1
oC. Sau 24 gi, lc nh ng v kim tra s to kh, tc l
s si bt. Li thm 24 gi nu quan st khng thy kh
thot ra. Kim tra kh ln 2. Thc hin kim tra xc nhn kt
qu c cho l dng tnh nh sau :
Lc nh ng LST c thot kh v ly chuyn mt que cy
ca th vn vo mt ng cha 10 ml canh trng EC v
mt l ln men (Durham). ng EC trong 24 48 gi
45,5 2 oC. Sau 24 gi kim tra s to kh; nu m tnh,
kim tra li sau 48 gi. Dng que cy ly mt vt ca th
vn t ng c sinh kh cho ln trn agar L-EMB. Quan trng
l mt phn ca mt thch xut hin nhng khun lc tch
bit. trong 18 24 gi 350C. Kim tra cc a i vi
cc khun lc in hnh ca E.coli tc l hng tm mu
thm c hoc khng c nh kim loi. Chn hai khun lc
c cho l dng tnh v chuyn chng vo thch
nghing PCA kim tra ho sinh v hnh thi hc. thch
nghing PCA trong 18 24 gi 35 1 oC, sau nhum
mu Gram trn chng nui cy. Nu mi trng Gram m
42
QCVN 4 - 21: 2011/BYT

(dng hnh que ngn) thc hin mt trong hai s th ho
sinh sau y:
S 1.A. Th hot tnh ho sinh IMViC:
a. Tnh sinh indol: Cy vo mt ng canh trng trypton v
trong 24 gi 350C. Th idol bng cch thm 0,2 -0,3 ml
ha cht Kovacs. Mu th dng tnh nu xut hin mu
khc bit lp ch lng pha trn.
b. Hp cht phn ng Voges-Proskauer: Cy vo mt ng
canh trng MR-VP v trong 48 gi 350C. Chuyn 1 ml
vo 1 ng c 13x100 mm. Thm 0,6 ml dung dch alphanaphthol v 0,2 ml dung dch KOH 40%, lc u. Thm vi
tinh th creatin. Lc v yn 2 gi. Mu th dng tnh
nu mu hng eosin lan trong mi trng.
c. Hp cht phn ng methyl: ng MR-VP t php th
Voges-Proskauer thm 48 gi 350C. Thm 5 git dung
dch methyl vo mi ng. Mu th dng tnh nu c
mu khc bit. Mu th m tnh nu c mu vng xut
hin.
d. Kh nng s dng citrat: Cy nh nhng vo mt ng
canh trng Koser citrat; trnh lm c. 350C trong 96
gi. Phn ng dng tnh khi c khc bit tng ln.
S 1.B. S dng chng cy mc trn thch nghing
PCA, cy chuyn vo 1 ng canh trng LST c cha l
Durham v 350C trong 48 gi kim tra s phn lp
c kh nng to acid v kh t qu trnh ln men lactose.
Ni suy: Cc chng cy (a) sinh kh l kt qu ca qu trnh
cy canh trng LST v tip sau trong 24 28 gi
350C, (b) xut hin nh l Gram m dng que khng sinh
bo t v (c) a cc dng IMViC (typ sinh hc 1) hoc +- (typ sinh hc 2) c xem l E.coli.
S 2. Phn tn bt k khun lc ang pht trin vo mt
th tch nh nc mui 0,85%. Khng nh nh tnh s
pht trin ca vi khun bi cc xt nghim ha hc c
thc hin thun tin bng cch s dng API 20E hoc di
Micro ID hoc cc h thng tng ng. Sau khi hon
thnh cc th nghim, xc nh nh tnh cc sinh vt t
Sch ch dn nh tnh ca h thng s dng v ghi li kt
qu cui cng.
43
QCVN 4 - 21: 2011/BYT

Salmonella
S dng cellulase phn r mu gm trc khi phn tch

l cn thit trnh to gel ca gm trong sut qu trnh
thm gm vo canh thang tng sinh. Chun b dung dch
cellulase 1% (1 g cellulase trong 99 ml nc) v tit trng
bng cch lc qua mng 0,45 m (Dung dch cellulase c
th bo qun 2 50C trong 2 tun). B sung 25 g mu
gm vo cc v trng dung tch 250 ml hoc dng c cha
khc thch hp. Cho vo 1 lp np xoy ming rng v
trng hoc 1 dng c chia khc thch hp 225 ml canh
trng lactose tit trng v 2,25 ml dung dch cellulase 1%
tit trng. Trong khi khuy canh trng cellulase/lactose
bng my khuy t cho nhanh 25 g mu qua phu thu tinh
v trng vo canh trng cellulase/lactose. y np dng
c cha chc chn v yn 60 pht nhit phng. Ni
lng np y v dng c cha 35 1 oC trong 24 2
gi. Sit cht np y v lc nh hn hp mu ; chuyn
1 ml hn hp vo 10 ml canh trng selenit cystin (SC) v 1
ml khc ca hn hp cho vo 10 ml canh trng
tetrathionat (TT). trong 24 2 gi 35oC. Lc (bng my
Vortex nu s dng ng) v ria 1 vng que cy 3 mm canh
trng TT trn agar bismuth sulfit (BS), agar xylose
lysin desoxycholat (XLD) v agar Hektoen enteric (HE).
(Chun b cc a BS mt ngy trc khi ria cy v bo
qun trong bng ti nhit phng). Lp li i vi canh
trng SC vi 1 vng 3 mm ca que cy. trong 24 2 gi
35oC. Tip tc nh c ch ra Quyn 4, "Kim tra cc
a v s hin din ca khun lc".
44
QCVN 4 - 21: 2011/BYT

Nm men v nm mc
Cn 25 g mu v thm vo 2.475 ml nc pepton 0,1%

tit trng (c chun b bng cch cho 1 g pepton vo 1 lt
nc ct, khuy u cho pepton tan hon ton v hp tit
trng 1210C trong 15 pht) trong khi khuy mnh bng
my khuy t. Khuy cho n khi mu c tan hon ton.
T l pha long 1 : 1.000. Dng pipet tit trng ly 0,1 ml
cho vo 10 a CCPDA-D c c lm trc mi
tm 0,1 ml (xem di y). Dn u ln b mt ca cc a,
s dng que thu tinh v trng. a thng ng trong 5
ngy 250C, yn, khng dao ng.
Sau khi , m khun lc pht trin trn mi a s dng
thit b m khun lc v ghi tng s khun lc c trn 10
a. Tch nm men ra khi nm mc da vo hnh thi ca
chng v m ring. Ly tng s khun lc c trong c 10
a nhn vi 100 c c CFU/g mu. Nu khng c
khun lc no trn a th biu din kt qu nh hn 100
CFU/g.
Mi trng CCPDA-D: Trc tin chun b dung dch gc
dichloran 2% (2,6-dichloran-4-nitroanilin) trong ethanol 95%.
Sau , thm mt lng dung dch gc dichloran vo
PDA t c nng 2,5 mg/l. Thm chloramphenicol
cho n nng 50 mg/l v hp tit trng 1210C trong 15
pht. Lm ngui mi trng n khong 500C trc khi rt
a, thm chlortetracyclin c lc v tit trng c
nng 50 mg/l.
45
QCVN 4 - 21: 2011/BYT

Ph lc 9
YU CU K THUT V PHNG PHP TH I VI GM GUAR
1. Tn khc, ch s
Gm cyamopsis, guar flour

INS 412
2. nh ngha
Xut x l ni nh c nghin ca cc ht t cy
Cyamopsis tetragonolobus (L) Taub. (Fam. Leguminosae)
ch yu bao gm polysaccharid c trng lng phn t
cao (khong 50.000 8.000.000) gm c cc
galactomannan; t l mannose: galactose l 2 : 1. Gm
c lm sch bng cch ra vi ethanol hoc
isopropanol hoc ho trong nc si, sau lc, c v
sy kh.
M s C.A.S.
9000-30-0
3. Cm quan
Dng bt ri mu trng n mu trng vng, gn nh

khng c mi
4. Chc nng
Cht lm dy, cht n nh, cht nh ho
5. Yu cu k thut
5.1. nh tnh
To gel
Thm lng nh dung dch natri borat TS vo dung dch

mu c lm phn tn bng nc; gel c hnh thnh.
nht
Phi c phn ng c trng ca nht.
Thnh phn gm
Kim tra hin vi
Nghin mu vi dung dch nc cha 0,5% iod v 1% kali

iodid, pht ln tiu bn knh v soi bng knh hin vi.
Gm guar cha cc t bo dng ht hnh trn hoc hnh tri
l, t thnh m, bn trong cha cc cht c mu vng n
nu. (Gm u carob gm c cc nhm t bo dng ng
cng di, tch bit hoc c khong trng nh. Cc thnh
phn bn trong ca gm u carob c mu nu c hnh
thnh t hn so vi trong gm Guar).
5.2. tinh khit

Gim khi lng khi
sy kh
Khng c qu 15,0% (nhit sy 1050C trong 5 gi).
Borat
Khng pht hin c.
Tro ton phn
Khng c qu 1,5%
Cht khng tan trong

acid
Khng c qu 7,0%
Protein
Khng c qu 10,0%.
46
QCVN 4 - 21: 2011/BYT

Khng c qa 1%, dng n cht hoc hp cht

(m t trong phn Phng php th)
Ch
5.3 Cc yu cu v vi
sinh vt
Tng s vi sinh vt
E. coli
m tnh i vi mu th
Salmonella
m tnh i vi mu th
Nm men v nm mc
Khng c qu 500 CFU/g
6. Phng php th
6.1. nh tnh
nht
Cho 2 g mu vo trong cc dung tch 400 ml v lm m

hon ton bng 4 ml isopropanol. B sung 200 ml nc v
khuy mnh, tip tc khuy cho n khi gm c phn tn
u hon ton. Mt dung dch hi nht mu trng sa c
hnh thnh. Chuyn 100 ml dung dch ny vo mt cc khc
dung tch 400 ml. Gia nhit hn hp trong cch thy si
trong 10 pht v lm ngui ti nhit phng. Khng c s
tng nht ng k (y l s khc bit gia gm guar
vi gm u carob ).
Thnh phn gm
Tin hnh nh hng dn phn nh tnh thnh phn

gm (FNP 5) s dng 100 mg mu thay th cho 200 mg v
1 10 l cht thy phn thay th cho 1 5 l. S dng
galactose v mannose nh l cht chun i chng. Cc
thnh phn ny cn phi c mt.
6.2. tinh khit

Borat
Ho tan 1 g mu trong 100 ml nc. Dung dch phi gi

c dng lng v khng to gel khi yn. Trn vo 10
ml dung dch acid HCl long v cho mt git hn hp va
trn ln trn giy ngh. Khng c to thnh nu m
dn trong khi sy v thay i thnh mu en lc khi lm m
bng dung dch ammoniac TS.
Protein
Tin hnh nh hng dn phn nh lng nitrogen

(Phng php Kjeldahl; FNP5). % protein trong mu bng
% nitrogen trong mu nhn vi 6,25.
47
QCVN 4 - 21: 2011/BYT

Nguyn tc:
c xc nh bng sc k kh (Xem quyn 4).
Chun b mu:
Ho tan 100 mg mu trong 10 ml nc s dng NaCl nh l
cht lm phn tn nu cn.
Dung dch ni chun:
Chun b dung dch nc cha 50 mg/l n - propanol
isopropanol
Cch tin hnh:
Cn 200 mg ure cho vo mt l sm mu dung tch 25 ml
Reacti-flasks, Pierce, Rockford, IL, USA, hoc tng
ng). Lm trong bng nitrogen trong 5 pht v sau
thm 1 ml dung dch acid oxalic bo ho, ng bng nt cao
su v lc xoy. Thm 1 ml dung dch mu, 1 ml dung dch
ni chun v ng thi bt u bm gi (T = 0). Lc xoy l
v y nt li bng nt xoy c m cao su silicon. Lc xoy
cho n khi T = 30 giy. Ti thi im T = 45 giy, bm qua
m cao su silicon 0,5 ml dung dch natri nitrit (250 g/l). Lc
mnh cho n khi T = 70 giy v T = 150 giy ht qua
m cao su silicon 1 ml khng gian hi s dng syringe
kho p (Precision Sampling Corp., Baton Rouge,
Louisiana, USA, hoc tng ng).
Sc k kh:
a kim syringe vo cng bm; y mu sau m
syringe v bm mu.
Tnh kt qu:
cch so snh cc din tch pic vi cc pic thu c nh sc
k kh khng gian hi c hnh thnh do thay th 1 ml
dung dch chun alcol cho 1 ml dung dch mu trong cch
tin hnh nu trn.
48
QCVN 4 - 21: 2011/BYT

Ch

5.3. Cc yu cu v vi
sinh vt
Tng s vi sinh vt

m phosphat v dng Stomacher, my lc hoc my khuy
ho tan hon ton. Gii hn thi gian ho tan khong 10
pht v sau dng pipet ly 1 ml dung dch vo cc a
petri ring bit, cp i v nh du thch hp. vo mi
a petri 12 15 ml agar trc c lm ngui n 44
460C. Trn u bng cch quay ln lt v di chuyn v
pha trc, pha sau cc a, cho agar ng c. o
ngc cc a v trong 48 2 gi 35 1 oC.
Sau khi m cc khun lc pht trin nhn thy c trn
hn 100 CFU/g.
E. coli
S dng cellulase phn gii mu gm trc khi phn tch

thm gm vo canh trng tng sinh. Chun b dung dch
th bo qun 2 50C trong 2 tun) cho vo ng tit trng
c cha 9 ml canh trng c lauryl sulfat tryptose (LST) v
trng, thm vo 0,1 ml dung dch cellulase 1% tit trng. B
sung 1 g mu gm vo ng v lc mnh bng my lc
Vortex phn tn u mu. ng trong 24 48 gi 35
1 oC. Sau 24 gi, lc nh ng v kim tra s to kh tc l
s si bt. Li thm 24 gi nu quan st khng thy kh
thot ra. Kim tra kh ln 2. Thc hin kim tra xc nhn kt
qu c cho l dng tnh (khi c kh c sinh ra) nh
sau:
Lc nh ng LST c bt kh v chuyn ly mt que cy ca
th vn vo mt ng cha 10 ml canh trng EC v mt l
ln men (Durham). ng EC trong 24 48 gi 45,5 2
o
C. Sau 24 gi kim tra s to kh; nu m tnh, kim tra li
sau 48 gi. Dng que cy ly mt vt ca th vn t ng c
sinh kh ria trn agar L-EMB. Quan trng l mt phn ca
mt thch xut hin nhng khun lc tch bit. trong 18
24 gi 350C. Kim tra cc a i vi cc khun lc in
hnh ca E.coli tc l hng tm mu sm c hoc khng
c nh kim loi. Chn hai khun lc c cho l dng tnh
v chuyn chng vo thch nghing PCA kim tra ho
sinh v hnh thi hc. thch nghing PCA trong 18 24
49
QCVN 4 - 21: 2011/BYT

o
gi 35 1 C, sau nhum mu Gram chng cy.

Nu chng cy Gram m (dng hnh que ngn) thc hin
mt trong hai s th ho sinh sau y:
S 1.1. Th hot tnh ho sinh IMViC:
a. To indol: Cy vo mt ng canh trng trypton v
trong 24 gi 350C. Th idol bng cch thm 0,2 -0,3 ml
ha cht Kovacs. Mu th dng tnh nu xut hin mu
khc bit lp cht lng pha trn.
b. Hp cht phn ng Voges-Proskauer: Cy vo mt ng
canh trng MR-VP v trong 48 gi 350C. Chuyn 1 ml
ti ng 13x100 mm. Thm 0,6 ml dung dch alpha-naphthol
v 0,2 ml dung dch KOH 40%, lc u. Thm vi tinh th
creatin. Lc v yn 2 gi. Mu th dng tnh nu mu
hng eosin lan trong mi trng.
c. Hp cht phn ng methyl: ng MR-VP t php th
Voges-Proskauer thm 48 gi 350C. Thm 5 git dung
dch methyl vo mi ng. Mu th dng tnh nu c
mu khc bit. Mu th m tnh nu c mu vng xut
hin.
d. Kh nng s dng citrat: Cy nh nhng mt ng canh
trng Koser citrat; trnh lm c. 350C trong 96 gi.
Phn ng dng tnh khi c khc bit tng ln.
Phng n 1.2. S dng chng cy ln thch nghing PCA,
cy li ng canh trng LST c cha l Durham v
350C trong 48 gi kim tra chng phn lp c kh nng
to acid v kh t qu trnh ln men lactose.
Ni suy: Cc chng cy (a) sinh kh l kt qu ca qu trnh
cy canh trng LST v sau trong 24 28 gi 350C, (b)
xut hin nh l Gram m dng que khng sinh bo t v
(c) cho cc dng IMViC (typ sinh hc 1) hoc -+- (typ sinh
hc 2) c xem l E.coli.
S 2. Phn tn bt k khun lc ang pht trin vo mt
th tch nh nc mui 0,85%. Khng nh nh tnh cc vi
khun mc bi cc xt nghim ha hc c thc hin
thun tin bng cch s dng API 20E hoc di Micro ID
hoc cc h thng tng ng. Sau khi hon thnh cc
th nghim, xc nh nh tnh cc sinh vt t Sch ch dn
nh tnh ca h thng s dng v ghi li kt qu cui cng.
Salmonella
S dng cellulase phn r mu gm trc khi phn tch

thm gm vo canh thang tng sinh. Chun b dung dch
th bo qun 2 50C trong 2 tun). B sung 25 g mu
gm vo cc v trng dung tch 250 ml hoc dng c cha
khc thch hp. Cho 225 ml canh trng lactose tit trng
v 2,25 ml dung dch cellulase 1% tit trng vo 1 l np
xoy, ming rng dung tch 500 ml v trng. Trong khi
khuy mnh canh trng cellulase/lactose bng my khuy
50
QCVN 4 - 21: 2011/BYT

t, cho nhanh 25 g mu qua phu thu tinh v trng vo
canh trng cellulase/lactose ny. y cht np l cha
chc chn v yn 60 pht nhit phng. Ni lng
np y v l cha 35 1 oC trong 24 2 gi. Sit cht
np y v lc nh hn hp mu ; chuyn 1 ml hn hp
vo 10 ml canh trng selenit cystin (SC) v 1 ml khc ca
hn hp cho vo 10 ml canh trng tetrathionat (TT).
trong 24 2 gi 35oC. Lc (bng my Vortex, nu s dng
ng) v ria cy 1 vng que cy (3 mm) canh trng TT
ln thch agar bismuth sulfit (BS), agar xylose lysin
desoxycholat (XLD) v agar Hektoen enteric (HE). (Chun b
cc a BS mt ngy trc khi ria cy v bo qun trong
bng ti nhit phng). Lp li i vi canh trng SC
bng cch ria mt vng cy 3 mm. trong 24 2 gi
35oC. Tip tc nh c ch dn trang 221 226 ca
Sch hng dn k thut, FAO Food and Nutrition Paper
s 5 ti bn ln 2, Rome 1991, "Kim tra cc a v s hin
din cc khun lc".
Nm men v nm mc
Cn 25 g mu v thm vo 2.475 ml nc pepton 0,1% tit

trng (c chun b bng cch cho 1 g pepton vo 1 lt
nc ct, khuy u cho pepton tan hon ton v hp tit
trng 1210C trong 15 pht) trong khi khuy mnh bng
my khuy t. Khuy cho n khi mu c tan hon ton.
T l pha long l 1 : 1.00. Dng pipet tit trng ly 0,1
ml cho vo 10 a CCPDA-D c c lm trc mi
a 0,1 ml (xem di y). Dn u ln b mt ca cc a,
s dng que thu tinh v trng. a thng ng trong 5
ngy 250C, yn.
Sau khi , m khun lc pht trin trn mi a s dng
thit b m khun lc v ghi tng s khun lc c trn 10
a. Tch nm men ra khi nm mc da vo hnh thi ca
chng v m chng ring. Ly tng s khun lc c trong
c 10 a nhn vi 100 c s CFU/g mu. Nu khng c
khun lc no trn cc a th biu din kt qu nh hn
100 CFU/g.
Mi trng CCPDA-D: Trc tin chun b dung dch gc
dichloran 2% (2,6-dichloran-4-nitroanilin) trong ethanol 95%.
Sau , thm mt lng dung dch gc dichloran va
vo PDA t c nng 2,5 mg/l. Thm
chloramphenicol cho n nng 50 mg/l v hp tit trng
1210C trong 15 pht. Lm ngui mi trng n khong
500C trc khi rt a, thm chlortetracyclin c lc v
tit trng c nng 50 mg/l.
51
QCVN 4 - 21: 2011/BYT

Ph lc 10
YU CU K THUT V PHNG PHP TH I VI GM TRAGACANTH
1. Tn khc, ch s
2. nh ngha
M s C.A.S.
INS 413
Thu c t vic sy kh dch tit ra ca thn v cnh cy
Astragalus gummifer Labillardiere v cc loi thuc Chu
khc ca Astragalus (Fam. Leguminosae); ch yu bao gm
cc polysaccharid c khi lng phn t cao (galactoarban
v polysaccharid c tnh acid), Gm Tragacanth thu phn
to ra acid galacturonic, galactose, arabinose, xylose v
fucose; mt lng nh rhamnose v glucose (c chuyn
ho t cc vt tinh bt hoc cellulose).
9000-65-1
3. Cm quan
Gm khng nghin dng mnh dt, l, phng hoc cong

hoc dng mnh xon c dy 0,5 2,5 mm v di ti 3
cm; mu trng ti vng nht, nhng mt s mnh c th c
mu nht; cc mnh cng, d gy nh; khng mi. Gm
dng bt mu trng cho n vng nht hoc mu nu hi
hng (mu nu vng nht).
Cc ch phm thng mi c th cha cc vt l nh mnh
v cy th phi loi b trc khi s dng cho thc phm.
Cc mu khng nghin phi c to thnh bt v cho qua
ry s 45 (335 M) v trn u trc khi tin hnh bt k mt
php th no sau y.
4. Chc nng
Cht nh ho, cht n nh, cht lm dy
5. Yu cu k thut
5.1. nh tnh
tan
Kim tra hin vi
To kt ta
Thnh phn gm
1 g mu trong 50 ml nc trng n to thnh dch c

nhy, trn, mu trng c; khng tan trong ethanol v
khng trng n trong ethanol 60% (w/v).
Xc nh bng knh hin vi huyn ph ca mu pha trong
nc. Nhiu mnh gc cnh vi cc dng l mng khng
u hoc trn, cc ht tinh bt ng knh ti 15 m v cc
mng t bo phn tng chuyn thnh mu tm nhn thy
c bng cch thm dung dch km clorid c cha iod.
Cc mu to ra phn ng kt ta vi dung dch ng (II)
acetat bo ho trong nc.
nh tnh arabinose, xylose, fucose, galactose v acid
galacturonic bng cch sau : Tin hnh nh hng dn
phn nh tnh thnh phn gm s dng cc cht chun
i chng : arabinose, mannose, galactose, xylose, fucose,
acid galacturonic v acid glucuronic. Arabinose, xylose,
fucose, galactose v acid galacturonic cn phi c trong
mu v mannose v acid glucuronic cn phi khng hin
din.
5.2. tinh khit

52
QCVN 4 - 21: 2011/BYT

Gim khi lng khi
sy kh
Khng c qu 16% (nhit sy 1050C trong 5 gi).
Tro sulfat
Khng c qu 4%
Tro khng tan trong

acid
Khng c qu 0,5%.
Cht khng tan trong

acid
Khng c qu 2%.
Gm acacia v cc
gm tan khc
t yu cu (m t trong phn Phng php th)
Agar
Dextrin
Gm karaya
Ch
5.3. Cc yu cu v vi
sinh vt
Salmonella spp.
m tnh trong 1 g mu th
E. coli
6. Phng php th
tinh khit
Tro khng tan trong
acid
un si tro thu c theo hng dn Tro sulfat trn vi

25 ml dung dch acid HCL 3M trong 5 pht, thu ly phn
cht khng tan ln trn mt chn lc cn b hoc giy lc
khng tro bit trng lng, ra bng nc nng, t v
cn. Tnh phn trm tro khng tan trong acid theo khi
lng mu.
Cht khng tan trong

acid
Cho 2 g gm tragacanth v 95 ml methanol vo 1 bnh y

trn dung tch 250 ml. Lm m bt bng cch lc xoy v
thm 80 ml dung dch acid HCl. Thm vi ht thu tinh
chng bt c ng knh khong 4 mm v un hi lu trong
cch thy trong 3 gi. Loi cc ht thu tinh chng bt ra v
lc ht chn khng trong khi nng qua phu lc thy tinh
xp bit trng lng. Ra bnh bng mt lng nh
nc v cho phn nc ra qua phu lc. Ra phn
cn trn phu lc bng 40 ml methanol v sy kh 1100C
ti trng lng khng i. ngui trong bnh ht m v
cn. Tnh phn trm cht khng tan trong acid theo khi
lng mu.
Gm acacia v cc
gm tan khc
Cho 10 ml dung dch Pb(C2H3O2)2 vo 20 ml dung dch mu

0,25% (khi lng/th tch) trong nc mi un si
53
QCVN 4 - 21: 2011/BYT

ngui. Kt ta thnh cm c to thnh. Lc v thm 10
ml dung dch Pb(C2H3O2)2. 2Pb(OH)2 vo dch lc. Dung
dch c th b vn c nh m khng to thnh kt ta.
Agar
Cho 0,5 ml dung dch acid HCl vo 4 ml dung dch mu

0,5% (khi lng/th tch) trong nc v gia nhit trong
cch thy si trong 30 pht. Thm vi git dung dch BaCl2
3,65% (khi lng/th tch). Khng c kt ta c hnh
thnh.
Dextrin
To mu trong dung dch nc glycerol v kim tra di

knh hin vi. B sung 1% dung dch iod, khng pht hin cc
ht mu vng nu hoc ta.
Gm karaya
(a) un si 1 g mu vi 20 ml nc cho n khi dch nhy

c hnh thnh. Thm 5 ml dung dch acid HCl v li un
si trong 5 pht. Khng c mu hng hoc mu bn l
r.
(b) Lc 0,2 g mu vi 10 ml ethanol 60% trong 1 ng ong
y np dung tch 10 ml c vch chia 0,1 ml. Bt k gel
no c hnh thnh cng khng c vt qu 1,5 ml.
(c) Lc 1 g mu vi 99 ml nc. Chun dch nhy c
hnh thnh bng dung dch NaOH 0,01M, s dng dung dch
methyl lm cht ch th. Khng qu 5,0 ml dung dch
NaOH 0,01M c dng lm thay i mu ca dung
dch.
Ch

54
QCVN 4 - 21: 2011/BYT

Ph lc 11
YU CU K THUT V PHNG PHP TH I VI GM ARABIC
1. Tn khc, ch s
2. nh ngha
M s C.A.S.
Gm arabic (Acacia senegal), gm arabic (Acacia seyal),

Acacia gum, arabic gum,
INS 414
Thu c t vic sy kh dch tit ra ca thn v cnh cy
Acacia senegal (L) Willdenow hoc Acacia seyal (fam.
Leguminosae).
Gm arabic ch yu bao gm cc polysaccharid c trng
lng phn t cao v cc mui Ca, Mg, K ca chng, khi
thu phn to to ra arabinose, galactose, rhamnose v
acid glucuronic. Cc ch phm thng mi c th gm c
cc cht l nh ct v cc mnh v cy, cc cht ny phi
c loi b trc khi cho vo thc phm.
9000-01-5
3. Cm quan
Gm arabic (Acacia senegal) l cht rn c mu trng

nht n nu da cam, khi b gy to cc khe gy trong.
Dng tt nht l dng ht hnh cu c cc kch c khc
nhau vi kt cu b mt sn si. Khi nghin, cc mnh c
mu trng nht hn v xut hin mu trng trong.
Gm arabic (Acacia seyal) gin hn gm arabic (Acacia
senegal) c cc ht dng hnh cu.
Gm arabic c sn trn th trng dng vy mu trng
n trng ng, dng ht, dng bt, dng ng hoc dng
nguyn liu c sy phun.
Dung dch nc 1 g trong 2 ml chy d dng v c tnh
acid khi th bng qu.
4. Chc nng
Cht nh ho, cht n nh, cht lm dy
5. Yu cu k thut
5.1. nh tnh
tan
Thnh phn gm
Quay quang hc
1 g ho tan trong 2 ml nc; khng tan trong ethanol

Tin hnh nh hng dn phn nh tnh thnh phn
gm (FNP 5) s dng cc cht chun i chng :
arabinose, galactose, mannose, rhamnose, acid
galacturonic, acid glucuronic v xylose. Phi c arabinose,
galactose, rhamnose v acid glucuronic; khng c cc vt
ph tng ng vi mannose, xylose v acid galacturonic.
Gm t A.senegal : dung dch tan trong nc quay tri
Gm t A.seyal : dung dch tan trong nc quay phi
Th mt dung dch mu, chun b bng cch cho 10 g
mu (ch phm kh) vo 100 ml nc (nu cn thit,
trc lc qua giy lc s 42 hoc phu lc c kch
thc l 0,8 m), s dng ng c chiu di 200 mm.
5.2. tinh khit
55
QCVN 4 - 21: 2011/BYT

Gim khi lng khi sy
kh
Khng c qu 15% (nhit sy 1050C trong 5 gi) i

vi dng ht v khng c qu 10% (nhit sy 1050C
trong 4 gi) i vi dng ch phm sy phun.
Cc mu cha nghin cn phi nghin thnh bt v qua
ry s 40, trn u trc khi cn.
Tro ton phn
Khng vt qu 4,0%
Tro khng tan trong acid
Khng c qu 0,5%
Cht khng tan trong

acid
Khng c qu 1,0%
Tinh bt hoc dextrin
Gm c tanin
Ch
5.3. Cc yu cu v vi
sinh vt
Salmonella spp.
m tnh i vi mu th
E. coli
6. Phng php th
Tinh bt hoc dextrin
un si dung dch mu 2%, lm ngui v thm vi git iod

TS. Khng c mu nh xanh hoc nh c hnh
thnh.
Gm c tanin
Thm 0,1 ml dung dch st (III) clorid TS vo 10 ml dung

dch mu 2%. Khng nhum mu en hoc to kt ta
en.
Ch

- Xc nh bng k thut quang ph 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.
56
QCVN 4 - 21: 2011/BYT

Ph lc 12
YU CU K THUT V PHNG PHP TH I VI GM XANTHAN
1. Tn khc, ch s
INS 415
2. nh ngha
Gm xanthan l polysaccharid c khi lng phn t cao

c sn xut bi qu trnh ln men cacbonhydrat vi
chng vi khun thun khit Xanthomonas campestris, lm
sch bng cch thu hi vi ethanol hoc isopropanol, sy
kh v nghin; c cha D glucose v D mannose l
cc n v hexose chim u th, cng vi acid D
glucuronic v acid pyruvic, v c x l nh mui Na, K
hoc Ca; cc dung dch ca gm xanthan trung tnh.
M s C.A.S.
11138-66-2
3. Cm quan
Dng bt mu kem.
4. Chc nng
Cht lm dy, cht n nh, cht nh ho, cht to bt
5. Yu cu k thut
5.1. nh tnh
tan
To gel
Tan trong nc; khng tan trong ethanol

5.2. tinh khit

0
Gim khi lng khi sy Khng c qu 15% (nhit sy 105 C trong 2,5 gi).
kh
Tro ton phn
Khng c qu 16% sau khi sy
Acid pyruvic
Khng c nh hn 1,5%
Xem m t trong phn Phng php th - nh tnh
Nitrogen
Khng c qu 1,5%
Tin hnh theo phng php Kjeldahl
Khng c qu 500 mg/kg, dng n cht hoc hp

cht
Xc nh theo m t phn Phng php th - nh tnh
Ch
Khng c qu 2 mg/kg.
Xc nh bng cch s dng k thut quang ph hp th
nguyn t ph hp vi tng mc . La chn lng mu
v phng php x l mu da vo cc nguyn tc ca
phng php c m t trong Quyn 4, Cc phng
php cng c
5.3. Cc yu cu v vi
sinh vt
Tng s vi sinh vt

57
QCVN 4 - 21: 2011/BYT

E.coli
m tnh i vi mu th
Salmonella
m tnh i vi mu th
Nm men v nm mc
Khng c qu 500 CFU/g

Xem m t di phn Phng php th - nh tnh
5.4. Hm lng
Hm lng tnh theo ch phm kh, khng c nh hn

4,2% v khng c qu 5,4% CO2, tng ng vi 91,0%
117,0% gm xanthan.
6. Phng php th
6.1. nh tnh
To gel
Cho 300 ml nc trc c un ti 800C vo cc

dung tch 400 ml v khuy nhanh bng my khuy, ti
thi im tc khuy t cc i cho hn hp kh gm
1,5 g mu v 1,5 g gm u carob. Khuy cho n khi hn
hp chuyn thnh dng dung dch v sau tip tc khuy
trong 30 pht na. Khng c nhit nc gim
xung di 600C trong qu trnh khuy. Ngng khuy v
hn hp ngui ti nhit phng trong t nht 2 gi.
Mt khi gel dai chc c hnh thnh sau khi nhit
gim xung di 400C, tuy nhin khng c gel nh vy
hnh thnh trong dung dch i chng 1% mu c chun
b theo cch tng t nhng khng c gm u carob.
6.2. tinh khit
58
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Acid pyruvic
Chun b mu :
Cn 600 mg mu chnh xc n 0,1 mg v ho tan vi
lng nc n 100 ml. Chuyn 10 ml dung dch vo 1
bnh thu tinh c nt mi dung tch 50 ml. Dng pipet ly 20
ml dung dch HCl N cho vo bnh, cn bnh v gia nhit
un si vi sinh hn ngc trong 3 gi, ch ngn tn
tht do bay hi. Lm ngui ti nhit phng v thm
nc cho b lng b tn tht trong qu trnh un. Dng
pipet ly 1 ml dung dch 2,4-dinitrophenylhydrazin 0,5%
trong acid HCl 2N cho vo phu tch dung tch 30 ml, sau
thm 2 ml dung dch mu, khuy u v nhit
phng trong 5 pht. Chit hn hp bng 5 ml ethyl acetat
v loi b lp nc. Chit hydrazon t ethyl acetat bng
ba phn 5 ml dung dch Na2CO3 TS, ly phn chit c
cho vo bnh nh mc dung tch 50 ml. Pha ti th tch 50
ml bng dung dch Na2CO3 TS v lc u.
Chun b chun:
Cn 45 mg acid pyruvic chnh xc n 0,1 mg v cho vo
bnh nh mc dung tch 500 ml. Pha ti th tch 500 ml
bng nc v lc u. Chuyn 10 ml dung dch ny vo
bnh thu tinh c nt mi dung tch 50 ml v tip tc theo
m t trong phn Chun b mu , bt u vi Dng
pipet ly 20 ml dung dch HCl N cho vo bnh .
Cch tin hnh :
o hp th ca mi dung dch vi my quang ph trong
cc cng o 1 cm bc sng c hp th cc i 375 nm,
s dng Na2CO3 TS cho mu trng. hp th ca
Dung dch mu bng hoc ln hp th ca Dung
dch chun .
59
QCVN 4 - 21: 2011/BYT

Nguyn tc:
c xc nh bng sc k kh khng gian hi (Xem quyn
4).
Chun b mu:
Ho tan 100 mg mu trong 10 ml nc s dng NaCl nh
l cht lm phn tn nu cn.
Dung dch ni chun:
Chun b dung dch nc cha 50 mg/l n - propanol
isopropanol
Cch tin hnh:
Cn 200 mg ure cho vo l sm mu dung tch 25 ml
Reacti-flasks, Pierce, Rockford, IL, USA, hoc tng
ng). Lm trong bng nitrogen trong 5 pht v sau
thm 1 ml dung dch acid oxalic bo ho, ng bng nt
cao su v lc xoy. Thm 1 ml dung dch mu, 1 ml dung
dch ni chun v ng thi bt u bm gi (T = 0). Lc
xoy l v y nt li bng nt xoy c m cao su silicon.
Lc xoy cho n khi T = 30 giy. Ti thi im T = 45
giy, bm qua m cao su silicon 0,5 ml dung dch natri
nitrit pha trong nc (250 g/l). Lc mnh cho n khi T =
70 giy v T = 150 giy ht qua m cao su silicon 1 ml
t khng gian hi s dng syringe kho p (Precision
Sampling Corp., Baton Rouge, Louisiana, USA, hoc
tng ng).
Sc k kh:
Lun kim syringe vo cng bm; y mu, sau m
syringe v bm mu.
Tnh kt qu:
cch so snh cc din tch pic vi cc pic tng ng thu
c t sc khng gian hi c hnh thnh do thay
th 1 ml dung dch chun alcol cho 1 ml dung dch mu
trong cch tin hnh nu trn.
60
QCVN 4 - 21: 2011/BYT

Ch

- Xc nh bng k thut quang ph 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. Cc yu cu v vi sinh
vt
Tng s vi sinh vt

m phosphat v dng Stomacher, my lc hoc my
khuy ho tan hon ton. Gii hn thi gian ho tan
khong 10 pht v sau dng pipet ly 1 ml dung dch
trn cho vo cc a petri ring bit, cp i v nh du
thch hp. vo mi a petri 12 15 ml thch PCA
trc c lm ngui n 44 460C. Trn u bng
cch quay o chiu di chuyn v pha trc, pha sau cc
a, cho agar ng li. Lt ngc cc a v trong 48
2 gi 35 1 oC.
hn 100 CFU/g.
E.coli
S dng k thut v trng, ho 1 g mu trong 99 ml canh

trng lactose, s dng hoc Stomacher, my lc hoc
my khuy cho ti khi mu tan hon ton. Gii hn thi
gian ho tan khong 15 pht v sau y nh dng c
cha v canh trng trong 18 24 gi 35 1 oC. Dng
1 pipet tit trng ly 1 ml canh trng cho vo ng
cha 10 ml canh trng GN. trong 18 24 gi v sau
ria cy cc ng canh trng GN cho thy sinh trng (+)
tnh hoc sinh hi ln cc a cp i Levine EMB. cc
a trong 24 2 gi 35 1 oC v sau kim tra cc
khun lc in hnh ca E.coli tc l biu th mu tm
m vo tm en v c nh kim loi xanh i lc ri rc
trn mt thch. Ghi li bt k khun lc E. coli in hnh
c cho l dng tnh, loi khc l m tnh. Ria cy bt
k khun lc kh nghi c phn tch r ln a PCA v
trong 18 - 24 gi 35 1 oC. Nhum mu Gram trn bt k
khun lc pht trin trn mi trng cy khng nh l
Gram m. Nu vy, phn tn bt k khun lc pht trin
vo lng nh nc mui 0,85% v tin hnh cc php
th ha hc nhn dng chng vi khun. iu ny c
lm thun li nht bng cch s dng di API 20E hoc
Micro ID hoc cc h thng tng ng.
Sau khi hon tt cc php th, nhn dng vi sinh vt theo
Hng dn nh danh ca h thng c s dng v ghi
li kt qu cui cng.
Cc mi trng nui cy
61
QCVN 4 - 21: 2011/BYT

Canh trng GN (Canh trng Gram m)
Pepton 20 g
Dextrose 1,0 g
Mannitol 2,0 g
Natri citrat 5,0 g
Natri deoxycholat 0,5 g
Dikali phosphat (hai baz) 4,0 g
Monokali phosphat (mt baz) 1,5 g
Natri chlorid 5,0 g
Pha thnh 1 l bng nc ct hoc nc kh ion, pH = 7,0
0,2 250C
Salmonella
S dng k thut v trng, phn tn 5 g mu trong 200 ml

canh trng lactose tit trng, s dng Stomacher, my
lc hoc my khuy lm ha tan ti a trn khong 15
pht. y lng dng c cha v 35 1 oC trong 24 2
gi.
Sit cht np y v lc nh hn hp mu ; chuyn 1
ml hn hp vo 10 ml canh trng selenit cystin (SC) v 1
ml khc ca hn hp cho vo 10 ml canh trng
tetrathionat (TT). trong 24 2 gi 35oC. Lc (bng my
Vortex nu s dng ng) v dng vng que cy 3 mm canh
trng TT ria ln agar bismuth sulfit (BS), agar xylose
lysin desoxycholat (XLD) v agar Hektoen enteric (HE).
(Chun b cc a BS mt ngy trc khi ria cy v bo
qun trong bng ti nhit phng cho n khi cy). Lp
li i vi canh trng TT vi 1 vng que cy. trong 24
2 gi 35oC. Tip tc nh c ch dn trang 221
226 ca Sch hng dn k thut, FAO Food and Nutrition
Paper s 5 ti bn ln 2, Rome 1991, "Kim tra cc a v
s hin din cc khun lc".
Nm men v nm mc

dung dch m phosphat v dng Stomacher, my lc hoc
my khuy ha tan hon ton. Gii hn thi gian ho
tan khong 10 pht v sau dng pipet ly 1 ml dung
dch cho vo cc a petri tch bit, cp i, nh du
tng ng. ln dch mu trong mi a petri 15 20 ml
Potato dextrose agar (hoc acid ho hoc cha cht
khng sinh) trc duy tr 44 460C. Trn u
bng cch quay o chiu v di chuyn trc sau cc a,
cho agar ng li. Lt ngc cc a v trong 5 ngy
20 - 25oC.
Sau khi , m cc khun lc mc nhn thy c trn mi
a, s dng thit b m khun lc v ghi tng s khun
lc. Tch ring nm men vi nm mc da vo hnh thi
v m chng ring. Ly trung bnh s khun lc c trn
hai a thch v nhn vi h s pha long 100. Nu khng
c khun lc no nhn thy trn a th biu din kt qu
nh hn 100 CFU/g.
62
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6.4. nh lng
Tin hnh theo hng dn trong phn th i vi Xc nh
CO2 bng cch kh carboxyl (Vol. 4), cn chnh xc 1,2 g
mu.
63
QCVN 4 - 21: 2011/BYT

Ph lc 13
YU CU K THUT V PHNG PHP TH I VI GM KARAYA
1. Tn khc, ch s
Gm Karaya, Sterculia, gm Sterculia, Kadaya, Katilo, Kullo,

Kuterra
INS 416
ADI "Khng gii hn"
2. nh ngha
Gm Karaya l nha kh thu c t thn v cnh ca cy

Sterculia urens Roxburgh v cc chi Sterculia khc (H
Sterculiaceae) hoc t Cocholospermum gossipium A.P. De
Candolle hoc cc chi khc ca loi Cocholospermum (H
Bixaceae).
Ch phm cha ch yu l cc acetyl polysaccarid cao phn
t, khi thy phn to ra acid galacturonic, galactose,
rhamnose cng vi lng nh acid glucuronic.
Ch s C.A.S
9000-36-6
3. M t
Ch phm th c dng git l vi cc kch thc khc nhau

v cc mnh v khng ng u c v ngoi dng bn tinh
th c trng; mu vng nht hoc nu hng, trong m;
cng v rp.
Dng bt gm mu xm nht n nh hng, c mi c
trng ca acid acetic. Ch phm thng mi c th c tp
cht (v cy) cn loi b trc khi s dng trong thc phm.
Ch phm th cn c nghin mn v qua ry chun ISO
c 45 (355m), trn u trc khi tin hnh kim nghim.
4. Chc nng
Cht lm dy, cht n nh, cht nh ha.
5. Tnh cht
5.1. nh tnh
tan
Ngm trng n 2g mu trong 50ml nc to ra gel dng

ht nhy, mu hi c, qunh; c tnh acid khi th bng qu;
khng tan trong ethanol.
Trng n trong dung

dch ethanol
Trng n trong ethanol 60%, tnh cht ny gip phn bit

gm Karaya vi cc loi gm khc.
To kt ta
Phi c phn ng to kt ta c trng.
Phn ng mu
Phi c phn ng mu c trng.
Cc thnh phn ca
gm
Tin hnh theo hng dn trong chuyn lun nh tnh cc

thnh phn ca gm ti JECFA monograph 1 - Vol.4, s
dng arabinose, galactose, mannose, rhamnose, xylose,
acid glucuronic v acid galacturonic lm chun i chiu.
Phi pht hin rhamnose, galactose, acid glucuronic v acid
galacturonic.
Khng c c mannose, arabinose, xylose.
64
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5.2. tinh khit
o
Gim khi lng khi lm Khng c qu 20,0% (sy ti 105 C trong 5 gi).
kh
Tro ton phn
Khng c qu 8,0%
Tro khng tan trong acid Khng c qu 1,0%.

Cht khng tan trong
acid
Khng c qu 3,0%.
Acid bay hi
Khng c thp hn 10,0% tnh theo acid acetic.
Tinh bt
Khng pht hin c.
Ch
5.3. Cc yu cu v vi
sinh vt
Salmonella spp.
m tnh trong 1g mu
E.coli
Khng c c trong 1g mu
6. Phng php th
6.1. nh tnh
To kt ta
Lc 1g mu th vi 80ml nc trong 24 gi. un si 4ml

dch nhy thu c vi 0,5ml acid hydrocloric c, thm 1ml
dung dch natri hydroxyd 5M, lc. Ly dch lc, thm 3ml
dung dch kali ng tartrat, un nng. Phi xut hin kt ta
.
Phn ng mu
un 1g mu th vi 20ml nc n khi to nhy, thm 5ml

acid hydrocloric, tip tc un hn hp trong 5 pht, phi xut
hin mu hoc hng bn.
Lm m 0,5g mu th vi 2ml natri hydroxyd 5M, phi xut
hin mu nu.
6.2. tinh khit

chn
Tro khng tan trong acid Cn 3g (chnh xc n 0,1 mg) mu th trong mt
nung cn b. Nung ti nhit thp (khong 550o), khng
c nung qu la, n khi tro ha hon ton, ngui
trong bnh ht m v cn. Nu tro cha sch carbon, tm
t tro, lc ly phn khng tan trn giy lc khng tro, cho
c giy lc v cn vo chn nung, nung n khi thu c tro
trng hoc gn trng. Gp tro vi phn dch lc v cho bay
hi n kh ri nung n thm. Nu tro vn khng
sch carbon, ngui chn nung, thm 15ml ethanol, dng
a thy tinh ph v khi tro, t ht ethanol v li nung n
thm, sau ngui. un si tro thu c vi 25ml
acid hydrocloric long (TS) trong 5 pht. Lc ly phn khng
tan trn phu Gooch cn b hoc giy lc khng tro, ra
65
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cn bng nc nng, nung, ngui v cn. Tnh hm
lng % tro khng tan trong acid theo khi lng mu
cn.
Cht khng tan trong
acid
Cn 5g mu (chnh xc n 0,1mg) cho vo bnh nn hoc

cc thy tinh 250ml cha sn 100ml acid hydrocloric 5%
(kl/tt). y cc bng mt knh ng h hoc gn bnh nn
vi sinh hn hi lu. un nh cho n khi gm tan hon
ton (khong 3 gi). Lc dung dch qua 1phu lc thy tinh
xp c l 10-20 m. Ra cn vi ln bng nc nng n khi
dch ra khng cn acid (th bng giy pH). Sy kh phu
n khi lng khng i 105o, ngui v nhit
phng trong bnh ht m v cn. Tnh hm lng %.
Acid bay hi
Cn 1g mu cho vo bnh thy tinh 700ml c di, thm

100ml nc v 5ml acid orthophosphoric, yn trong vi
gi hoc n khi gm trng n hon ton. un hi lu nh
trong 2 gi; Ct li cun hi nc ly khong 800ml dch
ct, phn acid cn li khong 20ml, chun dch ct thu
c bng natri hydroxyd 0,1M dng ch th l dung dch
phenolphtalein (TS). Tin hnh lm song song mt mu
trng khng c gm. S chnh lch gia 2 ln chun
biu th lng kim cn thit trung ha acid bay hi.
Mi ml natri hydroxyd 0,1M tng ng vi 0,006 g acid
bay hi tnh theo acid acetic.
Tinh bt
Thm vo dung dnh mu th 1/10, vi git dung dch iod

(TS). Dung dch khng c c mu xanh xut hin.
Ch

66
QCVN 4 - 21: 2011/BYT

Ph lc 14
YU CU K THUT V PHNG PHP TH I VI GM TARA
1. Tn khc, ch s
Peruvian carob
INS 417
ADI "Khng gii hn"
2. nh ngha
Thu c bng cch nghin mn phn ni nh ca ht cy

Caesalpinia spinosa (H Leguminosae); gm ch yu l cc
polysaccarid phn t lng ln cu to ch yu t
galactomannans. Thnh phn chnh gm chui mch thng
cc tiu phn (1,4)-beta-D-mannopyranose gn vi cc tiu
phn alpha-D-galacto-pyranose theo cc lin kt 1-6; T s
mannose/galactose trong gm tara l 3:1. (Trong gm u
carob t s ny l 4:1 v trong gm guar l 2:1). Ch phm
thng mi cn c phn loi thm qua nht v gim
khi lng khi lm kh.
3. M t
Bt trng hoc gn nh trng, gn nh khng mi.
4. Chc nng
Cht lm dy, cht n nh.
5. Tnh cht
5.1. nh tnh
tan
Tan trong nc, khng tan trong ethanol.
To gel
nht
Phi c phn ng c trng ca nht.
Cc thnh phn ca gm
Kim tra cu trc hin vi
Phi c phn ng c trng khi kim tra cu trc hin vi.
5.2. tinh khit

Gim khi lng khi lm
kh
Khng c qu 15%.
Tro ton phn
Khng c qu 1,5%.
Tro khng tan trong acid
Khng c qu 2,0%.
Protein
Khng c qu 3,5%.
Tinh bt
Khng pht hin c.
Ch
6. Phng php th
6.1. nh tnh
To gel
Thm lng nh natri borat vo dung dch mu th trong

nc, to thnh gel.
67
QCVN 4 - 21: 2011/BYT

nht
Cn 2 g mu th vo cc 400 ml, tm t bng khong 4 ml

isopropanol. Thm 200 ml nc, va thm va khuy mnh
n khi gm phn tn hon ton to thnh dung dch c
nht trung bnh, c nh sa ( nht ca dung dch ny
km dung dch gm guar nhng nht hn dung dch gm
u carob khi chun b cng trong iu kin nh trn).
Chuyn 100 ml dung dch ny sang cc 400 ml khc, un
nng hn hp trong b cch thy nc si trong 10 pht v
lm mt n nhit phng. nht ca dung dch tng
mt cch r rt.
Cc thnh phn ca gm
Th theo hng dn ti JECFA monograph 1 - Vol.4 - Tin

hnh theo hng dn trong chuyn lun nh tnh xc thnh
phn ca gm, s dng galactose v mannose lm chun.
Phi pht hin thy galactose v mannose.
Kim tra vi phu
Cho mt t bt mu vo trong dung dch cha 0,5% iod v

1% kali iodid trn mt lam knh, tin hnh soi di knh hin
vi. Gm tara cha cc nhm t bo hnh tri l hoc trn c
mu vng n nu. (cc t bo gm guar c hnh dng
tng t nhng kch thc ln hn. Trong khi t bo ca
gm u carob hnh ng di, tch ri nhau hoc gia chng
c khe nh rt d phn bit vi gm tara.
6.2. tinh khit

Protein
- Th theo hng dn ti JECFA monograph 1 - Vol.4 - Tin

hnh theo hng dn trong chuyn lun Xc nh nitrogen
(phng php Kjeldahl).
- Hm lng % ca nitrogen xc nh c nhn vi 5,7
thu c hm lng % protein trong mu th.
Tinh bt
Thm vo dung dch mu th (1/10) vi git dung dch iod

(TS) . Dung dch khng c xut hin mu xanh lam.
Ch

68
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69
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Ph lc 15
YU CU K THUT V PHNG PHP TH I VI GM GELLAN
1. Tn khc, ch s
INS 418
2. nh ngha
Gm gellan l gm polysaccharid c khi lng phn t cao

c sn xut bi qu trnh ln men chng vi khun thun
khit Pseudomonas elodea trong mi trng carbohydrat,
lm sch bng cch thu hi isopropyl alcol, sy kh v
nghin; Polysaccharid c khi lng phn t cao ch yu
gm mt tetrasaccharid lp i lp li ca mt n v
rhamnose, mt acid glucuronic, v hai n v glucose, v
c thay th bng cc nhm acyl (Glyceryl v acetyl) nh
cc ester lin kt O-glycosidic. Acid glucuronic c trung
ho thnh mui hn hp ca K, Na, Ca v Mg. Gm gellan
thng cha mt lng nh nitrogen c trong cc hp cht
thu c t qu trnh ln men.
M s C.A.S.
71010-52-1
Khi lng phn t
Khong 500.000
3. Cm quan
Dng bt mu trng nht
4. Chc nng
Cht lm dy, cht to gel, cht n nh
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, to thnh dung dch nht; khng tan trong
ethanol
To gel vi ion calci
Phi c phn ng to gel vi ion calci c trng.
To gel vi ion natri
Phi c phn ng to gel vi ion natri c trng.
5.2. tinh khit

Gim khi lng khi sy
kh
Khng c qu 15,0% (nhit sy 1050C trong 2,5 gi).
Nitrogen
Khng c qu 3,0%
Isopropyl alcol
Khng c qu 750,0 mg/kg (m t phn Phng php

th)
Ch
5.3. Cc yu cu v vi
sinh vt
Tng s vi sinh vt
Khng c qu 10.000 khun lc/g
E. coli
m tnh i vi mu th
Salmonella
m tnh i vi mu th
70
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Nm men v nm mc
5.4. Hm lng
Khng c nh hn 3,3% v khng c qu 6,8% CO2

tnh theo ch phm kh.
6. Phng php th
6.1. nh tnh
To gel vi ion calci
Cho 1,0 g mu vo 99 ml nc v khuy trong 2 gi, s

dng my khuy c cnh khuy kiu chn vt. Dng pipet ly
mt lng nh dung dch ny cho vo dung dch CaCl2
10%. Gel dng si dai c hnh thnh ngay lp tc.
To gel vi ion natri
Cho 1,0 g mu vo 99 ml nc v khuy trong 2 gi, s

dng my khuy c cnh khuy kiu chn vt. Thm 0,5 g
NaCl, gia nhit n 800C ng thi khuy, v gi 800C
trong 1 pht. dung dch ngui n nhit phng. Gel
cng chc c hnh thnh.
6.2. tinh khit
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QCVN 4 - 21: 2011/BYT

Isopropyl alcol
Dung dch chun isopropyl alcol (IPA):

Cho 500,0 mg isopropyl alcol t tiu chun dng cho sc
k vo bnh nh mc dung tch 50 ml, pha long ti th tch
50 ml bng nc v lc u. Dng pipet ly 10 ml dung dch
ny cho vo bnh nh mc dung tch 100 ml, pha long n
th tch 100 ml bng nc v lc u.
Dung dch chun tert - butyl alcol (TBA):
Cho 500,0 mg tert - butyl alcol t tiu chun dng cho sc
k vo bnh nh mc dung tch 50 ml, pha long ti th tch
50 ml bng nc v lc u. Dng pipet ly 10 ml dung dch
ny cho vo bnh nh mc dung tch 100 ml, pha long n
th tch 100 ml bng nc v lc u.
Dung dch hn hp chun:
Dng pipet ly 4 ml mi dung dch chun IPA v TBA cho
vo bnh Erlenmeyer dung tch 125 ml, pha long ti 100 ml
bng nc v lc u. Dung dch ny gm khong 40 g/ml
mi loi isopropyl alcol v tert butyl alcol.
Chun b mu:
Cho 1 ml nh tng chng to bt thch hp nh DowCorning G-10 hoc loi tng ng vo 200 ml nc
c ng trong bnh chng ct y trn dung tch 1.000
ml. Thm vo 5 g mu v lc bnh trong 1 gi trn my lc
c xoy. Ni bnh vi ct phn on v ct khong 100 ml;
iu chnh nhit bt khng tro vo ct. Thm 4,0 ml
dung dch chun TBA vo dch ct c c Dung dch
mu th.
Cch tin hnh:
Bm 5 l dung dch hn hp chun vo thit b sc k kh
c trang b detector ion ho ngn la v ct c lm
bng thp khng r, kch thc 1,8 m x 2,3 mm, c nhi
Porapak QS 80/100 mesh hoc loi tng ng. Kh
mang l He vi tc dng 80 ml/pht. Nhit bung
bm mu 2000C, nhit ct 1650C v nhit detector
2000C. Thi gian lu ca isopropyl alcol khong 2 pht, v
ca tert butyl alcol khong 3 pht.
Xc nh din tch cc pic ca IPA v TBA v tnh h s p
ng f = AIPA/ATBA, trong AIPA l din tch pic ca isopropyl
alcol v ATBA l din tch pic ca tert butyl alcol.
Tng t, bm 5 l dung dch mu th v xc nh cc din
tch pic, ghi li din tch pic ca isopropyl alcol l aIPA v
din tch pic ca tert butyl alcol l aTBA.
Tnh kt qu: hm lng isopropyl alcol (mg/kg) trong mu
c tnh theo cng thc sau:
(aIPA x 4.000)/(f x aTBA x W)
Trong W l khi lng mu phn tch (g)
72
QCVN 4 - 21: 2011/BYT

Ch

6.3. Cc yu cu v vi sinh
vt
Tng s vi sinh vt

m phosphat v dng Stomacher, my lc hoc my khuy
ho tan hon ton. Gii hn thi gian ho tan khong 10
pht v sau dng pipet ly 1 ml dung dch vo cc a
petri ring bit, cp i v nh du thch hp. ln dch
mu trong mi a petri 12 15 ml agar trc c lm
ngui n 44 460C. Trn u bng cch quay o chiu di
chuyn v pha trc, pha sau cc a, cho agar ng
li. Lt ngc cc a v trong 48 2 gi 35 1 oC.
hn 100 CFU/g.
73
QCVN 4 - 21: 2011/BYT

E.coli
S dng k thut v trng, ho 1 g mu trong 99 ml canh

trng lactose, s dng hoc Stomacher, my lc hoc
my khuy cho ti khi mu tan hon ton. Gii hn thi
gian ho tan khong 15 pht v sau y nh dng c
cha v canh trng trong 18 24 gi 35 1 oC. Dng
1 pipet tit trng ly 1 ml canh trng cho vo ng
cha 10 ml canh trng GN. trong 18 24 gi v sau
ria cy cc ng canh trng GN cho thy sinh trng (+)
tnh hoc sinh hi ln cc a cp i Levine EMB. cc
a trong 24 2 gi 35 1 oC v sau kim tra cc khun
lc in hnh ca E.coli tc l biu th mu tm m vo
tm en v c nh kim loi xanh i lc ri rc trn mt
thch. Ghi li bt k khun lc E. coli in hnh c cho l
dng tnh, loi khc l m tnh. Ria cy bt k khun lc
kh nghi c phn tch r ln a PCA v trong 18 - 24
gi 35 1 oC. Nhum mu Gram trn bt k khun lc
pht trin trn mi trng cy khng nh l Gram m.
Nu vy, phn tn bt k khun lc pht trin vo lng nh
nc mui 0,85% v tin hnh cc php th ha hc
nhn dng chng vi khun. iu ny c lm thun li
nht bng cch s dng di API 20E hoc Micro ID hoc
cc h thng tng ng.
Sau khi hon tt cc php th, nhn dng vi sinh vt theo
Hng dn nh danh ca h thng c s dng v ghi li
kt qu cui cng.
Cc mi trng
Canh trng GN (Canh trng Gram m)
Pepton 20 g
Dextrose 1,0 g
Mannitol 2,0 g
Natri citrat 5,0 g
Natri deoxycholat 0,5 g
Dikali phosphat 4,0 g
Monokali phosphat 1,5 g
Natri chlorid 5,0 g
Pha thnh 1 l bng nc kh ion, pH = 7,0 0,2 250C
Salmonella
S dng k thut thanh trng, phn tn 5 g mu trong 200

ml canh trng lactose tit trng s dng Stomacher, my
lc hoc my khuy ho tan ti a trn khong 15 pht.
y lng dng c cha v 35 1 oC trong 24 2 gi.
Tip tc nh bng phng php trang 221 ca FNP
5/Rev.2 (1991). nh tnh c th tin hnh thun tin hn khi
s dng cc h thng API hoc Micro ID hoc h thng
tng ng.
Nm men v nm mc

dung dch m phosphat v dng Stomacher, my lc hoc
my khuy cho n khi tan hon ton. Gii hn thi gian
74
QCVN 4 - 21: 2011/BYT

ho tan khong 10 pht v sau dng pipet ly 1 ml dung
dch vo cc a petri tch bit, cp i, nh du tng
ng. ln dch mu trong mi a petri 15 20 ml Potato
dextrose agar (hoc acid ho hoc cha cht khng
sinh) trc khng ch n 44 460C. Trn u bng
cch quay o chiu di chuyn v pha trc, pha sau cc
a, cho agar ng c. Lt ngc cc a v trong 5
ngy 20 250C.
Sau khi , m khun lc pht trin nhn thy trn mi a
s dng thit b m khun lc v ghi tng s khun lc.
Tch nm men ra khi nm mc da vo hnh thi v m
chng ring. Ly trung bnh s khun lc c trn cc a
canh trng nhn vi h s pha long 100. Nu khng c
khun lc no nhn thy trn a th biu din kt qu l nh
hn 100 CFU/g.
6.4. nh lng
Tin hnh theo hng dn trong phn th i vi Xc nh
CO2 bng cch kh carboxyl trong Cc phng php
chung, (Vol. 4), s dng chnh xc 1,2 g mu.
75
QCVN 4 - 21: 2011/BYT

Ph lc 16
YU CU K THUT V PHNG PHP TH I VI PECTIN
1. Tn khc, ch s
2. nh ngha
INS 440
Pectin bao gm ch yu l cc ester ca acid
polygalacturonic c methyl ho mt phn v cc mui
natri, kali, calci, amoni ca chng; Pectin thu nhn c t
qu trnh chit cc loi nguyn liu thc vt n c vi
nc, thng l to hay qu c mi (citrus); ch c methanol,
ethanol v isopropanol c s dng lm cht kim ta,
trong mt s trng hp mt phn ca cc ester methyl c
th chuyn thnh cc dng amid bc 1 khi x l bng
amoniac trong mi trng kim. SO2 c th thm vo lm
cht bo qun.
Sn phm pectin thng mi thng c pha long vi
ng nhm mc ch chun ho. Ngoi cc loi ng,
pectin c th c trn vi cc loi mui dng cho thc phm
khi cn nhm kim sot pH v to ra cc c tnh mong
mun. Thng tin thng mi ca sn phm c ch r gi tr
pH, mc to gel, nht, mc ester ho v cc c
trng k thut.
M s C.A.S.
9000-69-5
3. Cm quan
Dng bt mu trng, hi vng, hi xm hoc hi nu
4. Chc nng
Cht to gel, cht lm dy, cht n nh, cht nh ho
5. Yu cu k thut
5.1. nh tnh
tan
Tan trong nc, khng tan trong methanol, ethanol v

isopropanol
Pectin
Phi c phn ng c trng ca pectin.
Nhm amid
Ch p dng i vi cc pectin c amid ho (m t

phn Phng php th)
5.2. tinh khit

Gim khi lng khi
sy kh
Khng c qu 12,0 % (sy 1050C trong 2 gi)
SO2
Khng c qu 50 mg/kg.
D lng dung mi
Khng vt qu 1% methanol, ethanol v 2-propanol dng

n cht hoc hp cht
Tro khng tan trong

acid
Khng vt qu 1,0%
Tng s cht khng

tan
Khng qu 3,0%.
76
QCVN 4 - 21: 2011/BYT

Nitrogen
Khng vt qu 2,5% sau khi ra bng acid v ethanol
Acid galacturonic
Khng c nh hn 65,0% tnh theo ch phm kh v

khng c tro.
Mc amin ho
Khng vt qu 25,0% tng s cc nhm carboxyl ca

pectin.
Ch
6. Phng php th
6.1. nh tnh
Pectin
Lm m 0,05 g mu bng 2 - propanol. B sung 50 ml nc

v khuy bng thit b khuy t. iu chnh pH n 12 bng
dung dch NaOH 0,5 mol/l v yn dung dch trong thi gian
15 pht. Lm gim pH xung 7 bng acid HCl 0,5 mol/l. B
sung nc ct cho n th tch 100 ml. Cho cc mu vo cc
cuvet thch anh 1cm nh sau:
Dung
Dung
Dung dch m
pH = 7,0 *)
Khng
c
enzym
Khng c mu
th
Mu th
dch
mu
th
Nc
dch
enzym
**)
0,5 ml
1,0 ml
1,0 ml
0,5 ml
1,5ml
0,5 ml
0,5 ml
1,0 ml
0,5ml
0,5 ml
*)
Ho tan 6,055 g tris (hydroxymethyl) aminomethane (v d:

TRIZMA Base, Sigma) v 0,147 g CaCl2.2H2O trong 1 lt
nc. iu chnh pH n 7,0 bng acid HCl 1 mol/l.
**)
Pha long enzym tinh khit pectate lyase theo t l 1 : 100
bng dung dch m c pH = 7.
Lc u cc dung dch v o hp th bc sng 235 nm
ti thi im 0 v 10 pht.
Kt qu:
A0 = hp th 0 pht = Mu th (Khng c enzym + Khng
c mu th)
A10 = hp th 10 pht = Mu th (Khng c enzym + Khng
c mu th)
Lng sn phm khng bo ho sinh ra tng ng vi thay
i hp th (A10 - A0). Gi tr ny phi ln hn 0,023. iu
ny phn bit pectin so vi cc loi gm khc, th hin s
khng thay i v hp th.
77
QCVN 4 - 21: 2011/BYT

Nhm amid
Cho 2 ml acid HCl m c v 50 ml ethanol 60% vo 0,5 g

mu, khuy u trong 20 pht. Sau chuyn vo ng lc
thu tinh xp ra 6 ln mi ln 10 ml hn hp HCl-ethanol
60%. Ho tan trong 100 ml nc ct; cho thm mt vi git
NaOH 0,1 mol/l thu c dung dch. Chuyn 4 ml ca
dung dch ny vo ng nghim (c ng knh trong 15,5 mm
v chiu di 146 mm). Thm 1 ml dung dch NaOH 5 mol/l v
lc u. Hn hp s to thnh dng gel. Lm y ng thu
tinh nh (c ng knh trong 7,8 mm v chiu di 79 mm)
bng 2,5 ml acid boric TS v vo bn trong ng nghim.
Bao kn bng parafilm v qua m nhit 30 0C. Trong
trng hp c mt ca cc nhm amid cht ch th chuyn t
mu sang mu xanh l cy v gii phng ra kh
amoniac.
6.2. tinh khit

SO2
Trn 100 g mu vo 500 ml methanol trong bnh tam gic y

trn dung tch 1.000 ml c ng ht kh v ni c bnh vi sinh
hn ngc. Chun b mt khp ni thu tinh kt ni t sinh
hn ti bnh thu hoc ng ch U cha 10 ml dung dch
hydrogen peroxyd 3% c trung ho nh c dung dch
methyl TS. Ni ng ht kh vi ngun carbon dioxyd hoc
nitrogen khng c oxygen, duy tr dng kh nhm to ra bt
kh u u. Ngay sau b dng c ui ht khng kh,
30 ml dung dch acid HCl (10 ml dung dch acid HCl m c
pha trong 20 ml nc) vo sinh hn ngc, v ngay lp tc
ni bnh thu hoc ng ch U. Gia nhit t t cho n khi
methanol bt u chy ngc v un methanol chy
ngc t t trong 2 gi. Tho b dng c ra, cho dung dch
methyl vo v chun hydrogen peroxyd bng dung dch
NaOH 0,01 mol/l. Mi ml dung dch NaOH 0,01 mol/l tng
ng vi 0,32 mg SO2.
78
QCVN 4 - 21: 2011/BYT

D lng dung mi
p dng Phng php I trong Quyn 4, Phng php

chung, cc thnh phn hu c
Dung dch gc chun: Cn 5 g mi loi methanol, ethanol v
2 propanol cho vo 500 ml nc trong bnh nh mc dung
tch 1.000 ml. nh mc n th tch 1.000 ml bng nc.
Dung dch ni chun: Cn chnh xc 5 g 2 butanol (Wchun)
cho vo 500 ml nc trong bnh nh mc dung tch 1.000
ml v nh mc ti 1.000 ml bng nc.
Dung dch trng: Khng nh lng mu trng
Mu: Bo qun mu ni kh ro v mt. Trn u mu
trc khi phn tch.
Cn chnh xc 1 g mu (Wmu) vo cc dung tch 100 ml v
trn vi 5 g sucrose. Cho vo bnh Erlenmeyer c thanh
khuy t, thm vo 95 ml nc v 1 ml dung dch ni
chun. Trong khi khuy nhanh, thm t t hn hp pectinsucrose. y np bnh v khuy trong 2 gi. Pectin phi
c ho tan hon ton. Cn chnh xc 1 g dung dch ny
(Mmu) vo l phn tch bng GC khng gian hi.
Dung dch hiu chnh: Dng pipet ly 2 ml dung dch gc
chun v 2 ml dung dch ni chun cho vo bnh nh mc
dung tch 200 ml v nh mc ti 200 ml bng nc. Cn
chnh xc 1 g dung dch ny (Mchun) cho vo l v c
dng cho phn tch bng GC khng gian hi.
Cch tin hnh:
Tip tc phn tch nh m t Quyn 4 D lng dung
mi, s dng cc iu kin a ra tr nhit gia nhit
mu nn l 700C, v nhit syringe nn l 800C.
Tnh kt qu:
Nng (%) ca mi d lng dung mi:
Rmu x W chun x Mchun
% dung mi = --------------------------------------------x 100
Rchun x Wmu x Mmu x 1.000
Trong :
Rmu l din tch pic tng i ca mu
Rchun l din tch pic tng i ca chun
Wmu l khi lng mu (g)
Wchun l khi lng dung mi dng cho dung dch gc
chun
Mmu l khi lng dung dch mu dng cho phn tch bng
GC
Mchun l khi lng dung dch hiu chnh dng cho phn
tch bng GC.
79
QCVN 4 - 21: 2011/BYT

Tng s cht khng tan
Sy giy lc si thu tinh 70 mm (GF/B (Whatman code 1821

070) trong t sy c qut 1050C trong khong 1 gi.
Chuyn giy lc vo bnh ht m c cha silicagel v
ngui. Cn giy lc (M1). Cn khong 1 g (S) mu cho vo
cc dung tch 250 ml. Thm 5 ml 2 propanol phn tn
mu. Trong khi khuy t, thm 100 ml dung dch NaOH 0,03
mol/l c cha 0,1% (theo khi lng) ethylen diamin tetraacetic acid (mui natri), c lc qua giy lc GF/B.
Khuy khong 30 pht nhit phng, sau gia nhit
n si (khng phi gia nhit nu c qu nhiu bt).
Lc chn khng dung dch nng qua giy lc si thu tinh v
d kit lc chn khng c 3 phu Hartley (70 cm), c tm chu
nhit. Ra cc 5 ln v lc nc ra bng 100 ml nc m
(khong 500C) c lc qua giy lc GF/B.
Sy giy lc cng vi cn 1050C trong 1 gi. Chuyn vo
bnh ht m cha silicagel v ngui. Cn giy (M2). Tnh
% tng s cht khng tan:
Tng s cht khng tan (%) = [(M2 M1)/S] x 100
80
QCVN 4 - 21: 2011/BYT

Acid galacturonic v
mc amid ho
Cn 5 g mu chnh xc n 0,1 mg v chuyn vo cc thch

hp. Khuy trong 10 pht vi hn hp 5 ml dung dch acid
HCl TS v 100 ml ethanol 60%. Chuyn vo mt ng lc
thu tinh xp (dung tch t 30 - 60 ml) v ra bng 6 ln 15
ml hn hp HCl v ethanol 60%, sau bng ethanol 60%
cho n khi dch lc khng cn clorid. Cui cng ra bng
20 ml ethanol, sy trong 2,5 gi trong t sy 1050C, lm
ngui v cn. Chuyn chnh xc 1/10 tng khi lng tnh
ca mu sy kh (tng ng vi 0,5 g mu gc cha
c ra) vo bnh nn dung tch 250 ml v lm m mu
bng 2 ml ethanol TS. Thm 100 ml nc ct un si
ngui, ng nt li v thnh thong lc cho n khi dung
dch hon chnh c hnh thnh. Thm 5 git
phenolphthalein TS, chun bng dung dch NaOH 0,1
mol/l v ghi li kt qu nh chun ban u (V1).
Thm chnh xc 20 ml dung dch NaOH 0,5 mol/l, y nt
li, lc mnh v yn trong 15 pht. Thm chnh xc 20 ml
dung dch acid HCl 0,5 mol/l v lc cho n khi mu hng
bin mt. Chun bng dung dch NaOH 0,1 mol/l n
mu hng nht vn tn ti sau khi lc mnh; ghi li gi tr
ny nh l chun x phng ho (V2). Chuyn ton lng
cc thnh phn trong bnh nn vo bnh chng ct dung tch
500 ml c by Kjeldahl v sinh hn lm mt bng nc, ng
dn a xung pha di b mt ca 150 ml hn hp ca
nc khng c CO2 v 20,0 ml dung dch acid HCl 0,1 mol/l
trong bnh thu nhn. Thm 20 ml dung dch NaOH 10% vo
bnh chng ct, gn kn cc ch kt ni v sau bt u
gia nhit cn thn trnh bt tro. Tip tc gia nhit n khi
ly c 80 120 ml dch ct. Thm mt vi git dung dch
methyl TS vo bnh thu nhn v chun acid d bng
dung dch NaOH 0,1 mol/l, ghi li th tch chun S
(ml). Tin hnh nh lng mu trng trn 20,0 ml dung dch
acid HCl 0,1 mol/l v ghi li th tch s dng B (ml).
chun amid = B - S
Cho chnh xc 1/10 tng khi lng tnh ca mu kh
(tng ng vi 0,5 g mu gc cha c ra) v lm m
bng 2 ml ethanol trong cc dung tch 50 ml. Ho tan pectin
trong 25 ml dung dch NaOH 0,125 mol/l. Khuy dung dch
trong 1 gi nhit phng. Chuyn ton lng dung dch
pectin c x phng ho vo bnh nh mc dung tch
50 ml v pha long n th tch 50 ml bng nc ct.
Chuyn 20 ml dung dch pectin pha long vo thit b c
chng ct v thm vo 20 ml dung dch Clark c cha 100 g
magnesi sulfat heptahydrat, 0,8 ml dung dch acid H2SO4
m c v nc ct n tng th tch 180 ml. Thit b
chng ct gm c thit b bc hi nc kt ni vi bnh y
trn c ni vi sinh hn. C hai thit b bc hi nc v
bnh y trn u c trang b o nhit.
81
QCVN 4 - 21: 2011/BYT

Chng ct bng cch gia nhit bnh y trn cha mu. Ly
15 ml dch ct tch ra u tin cho vo ng ong c vch nh
mc. Sau cung cp hi v tip tc chng ct cho n khi
ly c 150 ml dch ct cho vo cc dung tch 200 ml. Thm
ton lng 15 ml dch ct u tin v chun bng dung
dch NaOH 0,05 mol/l ti pH = 8,5 v ghi li th tch chun
A (ml).
Thc hin nh lng mu trng trn 20 ml nc ct. Ghi li
th tch dng A0 (ml). chun ester acetat l (A A0).
Tnh mc amid ho (bng % tng cc nhm carboxyl) theo
cng thc:
Tnh hm lng acid galacturonic (mg) theo cng thc:

S mg acid galacturonic thu c theo cch ny chnh l hm
lng trong 1/10 trng lng mu kh c ra. Tnh %
acid galacturonic theo m v khng c tro, nhn s mg thu
c vi 1.000/x, trong x l khi lng mu kh c
ra (mg).
Ch 1: Nu pectin c bit l loi khng c amid ho,
ch cn xc nh V1 v V2, cn B S c coi l bng 0.
Ch 2: i vi pectin t qu to hoc qu c mi (citrus) th
hiu s A A0 khng c ngha trong tnh ton hm lng
acid galacturonic v mc amid ho.
Ch 3: Nu mun, tnh ester ho (bng % tng cc nhm
carboxyl) theo cng thc:
Ch 4: Nu mun, tnh ester acetat (bng % tng cc

nhm carboxylic t acid galaturonic) theo cng thc:
Ch

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.
82
QCVN 4 - 21: 2011/BYT

Ph lc 17
YU CU K THUT V PHNG PHP TH I VI METHYL CELLULOSE
1. Tn khc, ch s
Cellulose methyl ether;

INS 461
ADI khng gii hn
2. nh ngha
Ether methyl ca cellulose, c sn xut t bt g hoc

bng bng cch x l vi kim, v methyl ha cellulose kim
bng methyl clorid. Ch phm thng mi c th c phn
loi qua nht.
Tn ha hc
Methyl ether ca cellulose; Cellulose methyl ether
M s C.A.S.
9004-67-5
Cng thc ha hc
[C6H7O2(OH)x(OCH3)y]n
trong
x = 1,00 n 1,55
y = 2,00 n 1,45
x+y = 3,00
(y = thay th)
Cng thc cu to
trong R = H hoc CH3

Khi lng phn t
n v cu trc khng th: 162,14

n v cu trc c tng thay th l 1,45: 182
n v cu trc c tng thay th l 2,00: 190
i phn t: t khong 20000 (n khong 100) n khong
380000 (n c ga tr khong 2000)
3. Cm quan
Dng ht mn, si nh, hoc bt, mu trng hoc trng nht,

khng c mi, ht m.
4. Chc nng
Cht nh ha, cht n nh, cht lm dy.
5. Yu cu k thut
5.1. nh tnh
tan
Trng n trong nc, to dung dch t trong n trng

c, nht, keo; khng tan trong ethanol, ether v cloroform;
tan trong acid acetic bng.
To bt
Phi c phn ng to bt c trng.

83
QCVN 4 - 21: 2011/BYT

To kt ta
5.2. tinh khit

Gim khi lng khi lm Khng c qu 10,0% (sy ti 105 C, trong 3 gi).
kh
pH
5,0 - 8,0 (dung dch 1/100).
Tro sulfat
Khng c qu 1,5 %. (Th theo hng dn ti JECFA

monograph 1 - Vol.4 - Phng php I, cn 1 g mu).
Ch
5.3. Hm lng
Khng c thp hn 25% v khng c qu 33% nhm

methoxyl
6. Phng php th
6.1. nh tnh
To bt
Lc mnh dung dch mu th 0,1%. Cn xut hin mt lp

bt. (Php th ny cho php phn bit natri carboxylmethyl
cellulose vi cc ether cellulose khc.
To kt ta
Thm 5 ml dung dch ng sulfat hoc nhm sulfat 5% vo 5

ml dung dch mu th 0,5%. Trong dung dch khng c
xut hin kt ta. Php th ny cho php phn bit natri
carboxylmethyl cellulose vi cc ether cellulose khc.
6.2. tinh khit

Ch

6.3. nh lng
Xc nh hm lng nhm methoxyl bng phng php Xc
nh nhm Ethoxyl v Methoxyl (xem JECFA monograph 1 Vol.4 )
84
QCVN 4 - 21: 2011/BYT

Ph lc 18
I VI METHYL ETHYL CELLULOSE
1. Tn khc, ch s
MEC;
INS 465
ADI khng gii hn
2. nh ngha
Hn hp ether ca cellulose, c sn xut t cellulose

bng cch x l vi kim, dimethyl sulfat v ethyl clorid; c
hai nhm methyl v ethyl c gn vi cc tiu phn
anhydroglucose bng lin kt ether. Sn phm thng mi
c th c phn loi qua nht.
Tn ha hc
Ethyl methyl ete ca cellulose.
M s C.A.S.
9004-69-7
Cng thc ha hc
[C6H7O2(OH)x(OCH3)y(OC2H5)z]n
trong
z = 0,57 0,8
y = 0,2 0,4
x = 3 (y +z)
(y + z = thay th)
Cng thc cu to
Trong R = H hoc CH2 hoc C2H5

Khi lng phn t
n v cu trc khng th: 162,14

n v cu trc c tng thay th 0,77: 181
n v cu trc c tng thay th 1,2: 190
i phn t: 30000 n 40000 (n ~ 200)
3. Cm quan
Dng bt mn hoc hnh que, mu hi vng, khng mi v d

ht m.
4. Chc nng
Cht nh ha, cht n nh, cht lm dy, cht to bt.
5. Yu cu k thut
5.1. nh tnh
tan
Trng n trong nc, to dung dch t trong n trng c,

nht, keo; khng tan trong ethanol.
To bt
85
QCVN 4 - 21: 2011/BYT

To kt ta
Mc thay th
Xc nh mc thay th bng sc k kh.
5.2. tinh khit

Gim khi lng khi lm
kh
Tro sulfat
Khng c qu 15% i vi dng que, khng c qu

10% i vi dng bt. (sy n khi lng khng i).
Ch
5.3. Hm lng
Khng c thp hn 3,5% v khng c qu 6,5% nhm

methoxyl (-OCH3), khng c thp hn 14,5% v khng
c qu 19,0% nhm ethoxyl (-OCH2CH3), khng thp hn
13,2% v khng c qu 19,6% tng cc nhm alkoxyl, tnh
theo methoxyl (theo lng kh).
Khng c qu 0,6 %.
6. Phng php th
6.1. nh tnh
To bt
Lc mnh dung dch mu th 0,1%. Cn xut hin mt lp

bt. (Php th ny cho php phn bit natri carboxylmethyl
cellulose vi cc ether cellulose khc, cc alginat v cc gm
t nhin.
To kt ta
Thm 5 ml dung dch ng sulfat hoc nhm sulfat 5 % vo 5

ml dung dch mu th 0,5 %. Trong dung dch khng c
xut hin kt ta. (Php th ny cho php phn bit cc ether
cellulose vi natri carboxylmethyl cellulose, gelatin, gm carob
bean v gm tragacanth).
6.2. tinh khit

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

- Cn 1 g mu.
Ch

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
Xc nh nhm ethoxyl (c th xc nh ring bit ethoxyl v

methoxyl bng sc k kh (Cobler, Samsel and Beaver,
Talanta, 9, 473, 1962)).
Thit b
Thit b dng xc nh nhm hydroxypropoxy c m t
ra trong hnh v di y. Bnh ct hai c (D), c cnh c
ni vi ct Vigreaux c bc bng giy nhm (E) v c gia
ni vi vi mt ng sc kh chy t c ti y bnh dn hi
86
QCVN 4 - 21: 2011/BYT

nc v nitrogen. B phn to hi nc (B), c ni vi ng
sc kh qua ng C. Sinh hn (F), c ni vi ct Vigreaux.
Bnh ct v b phn to hi nc c t trong b du (A),
c gn thit b iu nhit duy tr nhit 155 C v tc
gia nhit mong mun. Sn phm ct c thu vo bnh 150 ml
(G), hoc mt dng c cha ph hp khc.
Tin hnh
Chuyn khong 100 mg (cn chnh xc n 0,1 mg) mu th
c sy kh 105 C trong 2 gi, vo bnh ct. Thm vo
10 ml dung dch crom trioxyd (60 g trong 140 ml nc). t
b phn to hi nc v bnh un vo b du (ti nhit
phng) ngang vch trn cng ca dung dch crom trioxyd. Bt
u lm lnh nc qua ng sinh hn v sc kh nitrogen qua
bnh un vi tc 1 bt/giy. Nng nhit b du t nhit
phng ln 155 C trong thi gian khng t hn 30 pht, v
duy tr nhit ny n khi xc nh xong. Ct ly khong 50
ml dch ct. Tho sinh hn ra khi ct Vigreaux, ra bng
nc, thu dch ra vo bnh ng dch ct. Chun dch ra
v dch ct bng NaOH 0,02 N n pH =7,0, s dng my o
pH vi thang o m rng,(Ghi ch: C th s dng dung dch
phenolphtalein (TS) l ch th cho php chun , trong trng
hp ny s dng ng thi i vi tt c mu chun v mu
trng).
Ghi li th tch dung dch NaOH dng chun , Va.
Thm 500 mg natri bicarbonat v 10 ml dung dch acid sulfuric
long (TS). Sau khi ht kh carbon dioxyd bay ra, thm mt
gam kali iodid. ng np bnh, lc u ri yn trong bng
ti trong 5 pht. Chun lng iod to thnh bng dung dich
natri thiosulfat 0,02 N ti khi mt mu vng, xc nh im kt
thc chun bng cch thm vi git dung dch h tinh bt
(TS). Ghi li th tch natri thiosulfat s dng, Ya.
Tin hnh lm vi mu trng (ha cht), trong ch s dng
dung dch crom trioxyd trong qui trnh trn. T s gia th tch
natri hydroxyd (Vb) v th tch natri thiosulfat (Yb) dng
chun c gi l t s acid/ oxi ha K=Vb/Yb, cho
crom trioxyd trong qa trnh ct phi l hng s i vi tt c
cc th nghim.
Tin hnh lm vi mu trng (chun), trong thay mu th
bng 100 mg methyl cellulose (khng cha tp cht). T s
87
QCVN 4 - 21: 2011/BYT

gia th tch natri hydroxyd (Vm) v th tch natri thiosulfat
(Ym).
Tnh hm lng ethoxyl (mg) ca mu th, theo cng thc
sau:
trong
N1 = Nng chnh xc ca dung dch natri hydroxyd 0,02N
N2 = Nng chnh xc ca dung dch natri thiosulfat 0,02N
k = VbN1/YbN2
Ghi li phn trm ethoxyl, B%.
Xc nh hm lng methoxyl
Xc nh hm lng methoxyl cng ethoxyl (hm lng tng
alkoxyl) nh hng dn trong phn Xc nh nhm Ethoxyl v
Methoxyl. Sau tnh hm lng methoxyl nh sau:
trong
A = hm lng tng alkoxyl , biu din bng % ethoxyl.
B = hm lng % ethoxyl, c xc nh trn.
Xc nh hm lng tng alkoxyl (theo methoxyl)
Mi ml dung dch natri thiosulfat 0,1N tiu tn trong qu trnh
xc nh hm lng tng alkoxyl tng ng vi 0,517 mg
alkoxyl, tnh theo methoxyl.
88
QCVN 4 - 21: 2011/BYT

Ph lc 19
I VI NATRI CARBOXYMETHYL CELLULOSE
1. Tn khc, ch s
Sodium cellulose glucolate, NaCMC, CMC, gm cellulose,

natri CMC,
INS 466
ADI khng gii hn
2. nh ngha
c sn xut t cellulose bng cch x l vi kim v

acid monoclo acetic hoc mui natri ca n. Sn phm
thng mi c th c phn loi qua nht.
Tn ha hc
Mui natri ca carboxymethyl ether ca cellulose.
M s C.A.S.
9004-32-4
Cng thc ha hc
[C6H7O2(OH)x(OCH2COONa)y]n
trong
n l polyme ha
x = 1,50
2,80
y = 0,2 1,50
x + y = 3,0 (y l thay th)
Cng thc cu to
Trong R = H hoc CH2COONa

Khi lng phn t
Mt n v cu trc c thay th 0,2: 178,14

Mt n v cu trc c thay th 1,5: 282,18
i phn t: ln hn 17000 (n c gi tr khong 100)
3. Cm quan
Dng ht, bt mn hoc hnh que nh, c mu trng hoc

hi vng. D ht m, gn nh khng c mi.
4. Chc nng
Cht lm dy, cht n nh, tc nhn to huyn ph.
5. Yu cu k thut
5.1. nh tnh
tan
To dung dch keo nht trong nc, khng tan trong

ethanol.
89
QCVN 4 - 21: 2011/BYT

To bt
To kt ta
Phn ng mu
Phi c phn ng mu c trng.
5.2. tinh khit

Gim khi lng khi lm Khng c qu 12,0 % sau khi sy (sy ti 105 C n khi
lng khng i)
kh
pH
6,0
8,5 (dung dch 1 trong 100).
Natri
Khng c qu 12,4 % tnh theo ch phm lm kh.
Natri clorid
Khng c qu 0,5 % tnh theo ch phm lm kh (m

t trong phn Phng php th).
Glycolat t do
Khng c qu 0,4 % tnh theo natri glycolat tnh theo ch

phm lm kh (m t trong phn Phng php th).
thay th
Khng c nh hn 0,2 v khng c qu 1,5 (m t

trong phn Phng php th).
Ch
5.3. Hm lng
Khng thp hn 99,5% natri carboxymethyl cellulose, tnh

theo ch phm c lm kh.
6. Phng php th
6.1. nh tnh
To bt
Lc mnh dung dch mu 0,1%. Trong dung dch khng

c xut hin lp bt. Php th ny cho php phn bit
natri carboxylmethyl cellulose Vi cc ether cellulose khc,
cc alginat v cc gm t nhin.
To kt ta
Thm 5 ml dung dch ng sulfat hoc nhm sulfat 5% vo

5 ml dung dch mu th 0,5% Trong dung dch khng c
xut hin kt ta. (Php th ny cho php phn bit natri
carboxylmethyl cellulose vi cc ether cellulose khc,
gelatin, gm carob bean v tragacanth).
Phn ng mu
Va thm 0,5 g natri carboxylmethyl cellulose dng bt vo

50 ml nc va khuy u to s phn tn ng nht.
Tip tc khuy cho n khi to thnh dung dch trong. Ly 1
ml dung dch ny vo mt ng nghim nh, pha long bng
nc vi th tch tng ng, thm 5 git 1-naphtol (TS).
Nghing ng nghim v cn thn thm vo thnh ng 2 ml
acid sulfuric sao cho to thnh mt lp mng nm di. Mu
tm phi xut hin mt phn cch gia hai lp.
6.2. tinh khit

Natri clorid
Nung nng nh trn bp 5g mu (cn chnh xc n 0,1 mg)

trong chn platin hoc chn s, u tin trn ngn la nh
mu khng b nung, khi than ha hon ton tip tc nung
90
QCVN 4 - 21: 2011/BYT

mu trong l in 15 pht nhit 500 C. Lm ngui,
nghin nh tro thu c v chit vi ln bng nc m. Lc
dch chit vo bnh nh mc 500 ml, acid ha bng acid
nitric v nh mc ti vch. Xc nh hm lng NaCl trong
100 ml dch chit ny bng phng php Volhard, s dng
dung dch bc nitrat 0,02 N v amoni nitrat 0,02N.
Mi ml dung dch bc nitrat 0,02 N tng ng vi 1,169
mg NaCl. Tnh hm lng natri clorid theo cng thc sau:
% natri glucolat = (a x 0,001169 x 5) / b
trong a = Th tch (ml) dung dch bc nitrat 0,02 N
dng chun .
b = Khi lng mu th sau khi tr i hm lng nc (g).
91
QCVN 4 - 21: 2011/BYT

Glycolat t do
Cn 0,5g mu (chnh xc n 0,1 mg) ri chuyn vo mt

cc vi c m dung tch 100 ml. Cn trng thm t mu
bng 5 ml acid acetic bng, sau bng 5 ml nc ri
khuy bng a thy tinh n tan hon ton (thng mt
khong 15 pht). Va khuy va thm t t 50 ml aceton,
sau thm khong 1g natri sulfat. Tip tc khuy thm vi
pht m bo ton b carboxylmethy cellulose kt ta
ht. Lc qua giy lc mm, c thm t trc bng
mt lng nh aceton. Thu dch lc vo bnh nh mc 100
ml. Dng 30 ml acetone thc y qu trnh lc v ra
kt ta. nh mc ti vch bng aceton v lc u.
Chun b mu trng gm 5 ml nc, 5 ml acid acetic bng
v aceton trong bnh nh mc 100 ml khc. Dng pipet ly 2
ml mu th v 2 ml mu trng vo 2 bnh nh mc 25 ml.
Lm bay hi aceton bng cch un cch thy bnh nh
mc, khng ng nt trong 20 pht. Lm ngui n nhit
phng v thm vo 5 ml dung dich naphthalendiol (TS),
lc k, sau thm tip vo 15 ml Dung dch naphthalendiol
(TS) v lc u. y ming bnh bng mt mnh giy nhm
v un cch thy trong 20 pht. Lm ngui n nhit
phng v nh mc n vch bng dung dch naphthalendiol
(TS).
o hp th quang ca dung dch mu so vi mu trng
ti bc sng 540 nm s dng cuvet 1cm. c s mg acid
glycolic tng ng t ng chun nh sau:
Thm 0; 1; 2; 3; v 4 ml dung dch acid glycolic chun (1
mg/ml, c pha nh sau: cn chnh xc 0,100g acid
glycolic ( c lm kh trong bnh ht m chn khng t
nht 16h), sau ha tan vo 100 ml nc, khng gi dung
dch qu 30 ngy) vo 5 bnh nh mc 100 ml. Thm nc
vo mi bnh n th tch 5 ml, sau thm 5 ml acid acetic
bng v nh mc n vch bng aceton v lc u. Dng
pipet ly 2 ml ca mi dung dch (c cha ln lt 0; 1; 2; 3
v 4 mg acid glycolic trong 100 ml) vo 5 bnh nh mc 25
ml v tin hnh qui trnh nh miu t trong phn chun b
dung dch th.
Dng ng chun bng cch v th biu th s ph
thuc gia lng acid glycolic (mg) trong 100 ml dung dch
ban u vi hp th quang. Tnh hm lng natri
glycolat (glycolat t do) theo cng thc:
% natri glycolat = (a x 0,129) / b
trong
a = Lng (mg) glycolic acid tnh t ng chun
b = Khi lng mu th sau khi tr i hm lng nc (g).
thay th
Chun b mu
Cn 5g mu (chnh xc n 0,1 mg) v chuyn vo bnh nn
500 ml. Thm 350 ml methanol hoc ethanol (80% theo th
tch). Lc huyn ph trong 30 pht. Gn qua phu lc Thy
tinh cn b, ht chn khng nh. Cui qu trnh lc gn
92
QCVN 4 - 21: 2011/BYT

trnh khng khng kh b ht qua phu. Lp li qu trnh
trn vi dung mi chit cho n khi th ion clorid bng dung
dch bc nitrat (TS) cho kt qu m tnh. Thng cn x l
khong 3 ln. Chuyn dung dch natri carboxylmethyl
cellulose vo chn. Loi b dung mi chit cn bm trn
cn bng aceton . Sy kh chn trong l nhit 110 C
n khi lng khng i. Cn ln u sau 2 gi. Mi ln
cn phi lm ngui chn trong bnh ht m v trong khi cn
cn lu rng natri carboxylmethyl cellulose ht m nh.
Tin hnh
Cn 2g mu kh (chnh xc 0,1 mg), thu c t qui trnh
chit cn trn vo chn s cn b. u tin nung nng
nh trn bp t chy mu t t, sau tng nhit
nung nng mnh trong 10 pht. Lm ngui, sau thm t
tro cn li bng 3-5 ml acid sulfuric c. Cn thn nung cho
ti khi ht khi bay ra. Sau khi lm ngui, thm khong 1 g
amoni carbonat, dn u bt trong ton b th tch ca chn.
Nung nng li trn bp, u tin nh nhng cho n khi ht
khi, sau nung n nng thm trong 10 pht. Lp li
qui trnh x l vi axid sulfuric v amoni carbonat nu tro
natri sulfat vn cn cha carbon. Lm ngui chn trong bnh
ht m v cn. Nu khng thm amoni carbonat v nung
trn bp, c th nung chn trong l trong 1 gi nhit
khong 600oC.
Tnh hm lng natri ca mu chit vi cn theo cng thc
sau:
% natri = (a X 32,28) / b
trong :
a = khi lng ca cn natri sulfat
b = khi lng cn kh thu c sau giai on chit bng
cn
Tnh thay th theo cng thc sau:
Mc thay th = (162 X % natri) / [2300-(80 X % natri)]
Ch

6.3. nh lng
Tnh hm lng % natri carboxymethyl cellulose trong mu

bng cch ly 100% tr i tng % ca natri clorid v natri
glucolat (glucolat t do), c xc nh theo hng dn
trn.
Hm lng (%) = 100 - (% NaCl + % Na glycolat)
93
QCVN 4 - 21: 2011/BYT

Ph lc 20
I VI GELATIN THC PHM
1. Tn khc, ch s
Gelatin edible
ADI khng gii hn
2. nh ngha
Gelatin thc phm c sn xut bng cch bn thy phn

collagen trong da, gn, dy chng, xng ca ng vt.
Ch phm thng mi c th c phn loi theo cc tiu
ch nh: bn ca gel, gii hn st, calci, lactose v cc
ha cht khc; cc gii hn i vi vi sinh vt gy bnh bao
gm Salmonella, Staphylococcus aureus, Clostridium spp.
v bo t nm mc. cc yu cu v gii hn vi sinh vt di
y l tm thi.
Ch s C.A.S.
9000-70-8
3. Cm quan
Dng phin, vy, mnh hoc bt mn, bt th, mu vng nht

hoc h phch, mu ty theo kch thc tiu phn, c mi
nh ging mi nc tht; bn trong khng kh kh nhng
trong khi ht m hoc trong dung dch b vi sinh vt lm
hng.
4. Chc nng
Cht to gel, cht n nh, cht nh ha, cht km hm cho

qu trnh kt tinh.
5. Yu cu k thut
5.1. nh tnh
tan
Khng tan trong nc lnh, nhng trng n v mm ra

ng thi ht nc vi khi lng gp 5-10 ln khi ngm
trong nc; tan trong nc nng, khi ngui to thch ng;
tan trong acid acetic; khng tan trong ethanol, cloroform v
ether.
To kt tu
To c
Phi c phn ng to c c trng.
Gii phng amoniac
Khi un nng mu th vi soda vi, kh amoniac s c

gii phng.
5.2. tinh khit

o
Gim khi lng khi lm Khng c qu 18% (sy ti 100 - 105 trong 6 gi).
kh
Cc hp cht khng tan

trong nc v c mi
Dung dch mu th 1/40 nng khng c c mi kh chu.

Khi quan st lp dy 2 cm, hu nh trong, ch c php
trng c nh sa rt nh.
Lu hunh dioxyd
Tro
Khng c qu 2,0%.
94
QCVN 4 - 21: 2011/BYT

Arsen
Ch
Cadimi
Thy ngn
5.3. Cc yu cu v vi
sinh vt
kh
< 104 /g
< 10/g
Enterobacteriaceae
hoc vi khun nhm coliaerogens
Streptococci nhm
Lancefield D
< 102 / g
6. Phng php th
6.1. nh tnh
To kt tu
Pha dung dch mu th 1/100, thm hn hp gm dung dch

trinitrophenol (TS) hoc dung dch kali dichromat 1/15 trn
vi acid hydrocloric long (t l trn 5/1 - tt/tt), trong dung
dch xut hin kt ta vng.
Pha dung dch mu th 1/100, thm dung dch thy ngn (II)
nitrat, trong dung dch xut hin kt ta trng, khi un nng
kt tu ny s chuyn thnh mu gch.
To c
Pha dung dch mu th 1/5000, thm dung dch acid tannic

(TS), dung dch s tr nn c.
6.2. tinh khit

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

- Phng php II.
Ch

Cadimi

- Phng php II.
Thy ngn

- Phng php II.
95

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CNG HO X HI CH NGHA VIT NAM

QCVN 4 - 21: 2011/BYT

QUY CHUN K THUT QUC GIA

QCVN 4 - 21: 2011/BYT

QCVN 4 - 21: 2011/BYT

QUY CHUN K THUT QUC GIA

QCVN 4 - 21: 2011/BYT

Yu cu k thut v phng php th i vi acid

Yu cu k thut v phng php th i vi kali

Yu cu k thut v phng php th i vi amoni

Yu cu k thut v phng php th i vi calci

Yu cu k thut v phng php th i vi propylen

Yu cu k thut v phng php th i vi agar

Yu cu k thut v phng php th i vi

Yu cu k thut v phng php th i vi gm u

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi gm

Yu cu k thut v phng php th i vi pectin

Yu cu k thut v phng php th i vi methyl

Yu cu k thut v phng php th i vi methyl

Yu cu k thut v phng php th i vi natri

QCVN 4 - 21: 2011/BYT

Yu cu k thut v phng php th i vi gelatin

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

3. Ly mu theo hng dn ti Thng t 16/2009/TT-BKHCN ngy 02 thng 6

QCVN 4 - 21: 2011/BYT

QCVN 4 - 21: 2011/BYT

Acid alginic t nhin thu c bng cch thy phn keo

Cng thc cu to do Phillips, Wedlock and Williams cng

Khi lng phn t

S lng v th t ca cc phn mannuronat v gluconat c

Dng ht, bt hoc si mnh mu trng n vng nu.

Cht n nh, cht lm dy, cht to gel, cht nh ha

Huyn ph mu th 0,3/10 c pH trong khong 2,0 - 3,5.

Phi c phn ng to kt ta vi amoni sulfat c trng.

Phi c phn ng c trng ca alginat.

5.2. tinh khit

QCVN 4 - 21: 2011/BYT

Khng c qu 8% tnh theo ch phm lm kh.

Cht khng tan trong

Khng c qu 2% tnh theo ch phm lm kh.

Khng c qu 3,0 mg/kg.

Khng c qu 5,0 mg/kg.

Khng c qu 5000 khun lc/g. Ban u chun b m

Khng c qu 500 khun lc/g

Ch phm khan cha khng thp hn 20,0% v khng c

Thm dung dch amoni sulfat bo ha vo dung dch mu

Ha tan 0,1 g mu th vi 0,15 ml dung dch natri hydroxyd

6.2. tinh khit

Cn khong 1 g (chnh xc n mg) mu th, ha tan trong

QCVN 4 - 21: 2011/BYT

Th theo JECFA monograph 1 - Vol.4. (phng php II)

- Th theo JECFA monograph 1 - Vol.4.

QCVN 4 - 21: 2011/BYT

Mui Kali ca acid alginic

Cng thc cu to do Phillips, Wedlock and Williams cng

S lng v th t ca cc phn mannuronat v gluconat c

n v cu trc : 214,22 (l thuyt); 238 (trung bnh thc t).

Dng ht, bt hoc si mnh mu trng n vng nu.