2

CNG BO/S 539 + 540/Ngy 27-10-2011

PHN VN BN QUY PHM PHP LUT

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 (Ban hnh km theo Thng t s 01/2011/TT-BYT ngy 13 thng 01 nm 2011 ban hnh cc Quy chun k thut quc gia i vi ph gia thc phm)

(Tip theo Cng bo s 537 + 538) Ph lc 12 YU CU K THUT V PHNG PHP TH I VI GM XANTHAN 1. Tn khc, ch s 2. nh ngha INS 415

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 ha, cht to bt 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc; khng tan trong ethanol To gel Phi c phn ng to gel c trng. 5.2. tinh khit Gim khi lng Khng c qu 15% (nhit sy 1050C trong 2,5 gi). khi sy kh

CNG BO/S 539 + 540/Ngy 27-10-2011

Tro ton phn Acid pyruvic Nitrogen Ethanol v isopropanol Ch

Khng c qu 16% sau khi sy Khng c nh hn 1,5% Xem m t trong phn Phng php th - nh tnh 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 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 E.coli Salmonella Nm men v nm mc

Khng c qu 5.000 CFU/g m tnh i vi mu th m tnh i vi mu th 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

6.2. tinh khit Acid pyruvic

Ethanol v isopropanol

Chun b mu: Cn 600 mg mu chnh xc n 0,1 mg v ha tan vi lng nc n 100 ml. Chuyn 10 ml dung dch vo 1 bnh thy 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 thy 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". Nguyn tc: Cc alcol c chuyn thnh cc ester nitrit tng ng v c xc nh bng sc k kh khng gian hi (Xem quyn 4). Chun b mu: Ha 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 Dung dch alcol chun: Chun b dung dch nc cha 50 mg/l mi loi ethanol v isopropanol

CNG BO/S 539 + 540/Ngy 27-10-2011

Ch

Cch tin hnh: Cn 200 mg ure cho vo l sm mu dung tch 25 ml Reactiflasks, Pierce, Rockford, IL, USA, hoc tng ng). Lm trong bng nitrogen trong 5 pht v sau thm 1 ml dung dch acid oxalic bo ha 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 kha 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. S dng cc iu kin sau: - Ct: thy 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 ha 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 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. - 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. Cc yu cu v 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

CNG BO/S 539 + 540/Ngy 27-10-2011

E.coli

ha tan hon ton. Gii hn thi gian ha 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. 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. S dng k thut v trng, ha 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 ha 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 Canh trng GN (Canh trng Gram m) Pepton 20 g Dextrose 1,0 g Mannitol 2,0 g

CNG BO/S 539 + 540/Ngy 27-10-2011

Salmonella

Nm men v nm mc

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 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". S dng k thut v trng, phn tn 1 g mu trong 99 ml dung dch m phosphat v dng Stomacher, my lc hoc my khuy ha tan hon ton. Gii hn thi gian ha 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 ha 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. 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.

6.4. nh lng

CNG BO/S 539 + 540/Ngy 27-10-2011

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" 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. 9000-36-6 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. Cht lm dy, cht n nh, cht nh ha.

2. nh ngha

Ch s C.A.S 3. M t

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 Trng n trong ethanol 60%, tnh cht ny gip phn bit gm Karaya vi cc loi gm khc. dung dch ethanol 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 Tin hnh theo hng dn trong chuyn lun nh tnh cc thnh phn ca gm ti JECFA monograph 1 - Vol.4, s dng ca gm

4. Chc nng 5. Tnh cht 5.1. nh tnh tan

CNG BO/S 539 + 540/Ngy 27-10-2011

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. 5.2. tinh khit Gim khi lng Khng c qu 20,0% (sy ti 105oC trong 5 gi). khi lm kh Tro ton phn Khng c qu 8,0% Tro khng tan Khng c qu 1,0%. trong acid Cht khng tan Khng c qu 3,0%. trong acid Acid bay hi Khng c thp hn 10,0% tnh theo acid acetic. Tinh bt Khng pht hin c. Ch Khng c qu 2,0 mg/kg. 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 Tro khng tan Cn 3g (chnh xc n 0,1 mg) mu th trong mt chn nung trong acid 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

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CNG BO/S 539 + 540/Ngy 27-10-2011

Cht khng tan trong acid

Acid bay hi

Tinh bt Ch

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 cn bng nc nng, nung, ngui v cn. Tnh hm lng % tro khng tan trong acid theo khi lng mu cn. 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 10-20m. 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 %. 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. Thm vo dung dnh mu th 1/10, vi git dung dch iod (TS). Dung dch khng c c mu xanh xut hin. - 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

11

Ph lc 14 YU CU K THUT V PHNG PHP TH I VI GM TARA 1. Tn khc, ch s 2. nh ngha Peruvian carob INS 417 ADI "Khng gii hn" 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. Bt trng hoc gn nh trng, gn nh khng mi. Cht lm dy, cht n nh.

3. M t 4. Chc nng 5. Tnh cht 5.1. nh tnh tan To gel nht Cc thnh phn ca gm Kim tra cu trc hin vi 5.2. tinh khit Gim khi lng khi lm kh Tro ton phn Tro khng tan trong acid Protein Tinh bt Ch

Tan trong nc, khng tan trong ethanol. Phi c phn ng to gel c trng. Phi c phn ng c trng ca nht. Phi c phn ng c trng ca thnh phn gm. Phi c phn ng c trng khi kim tra cu trc hin vi.

Khng c qu 15%. Khng c qu 1,5%. Khng c qu 2,0%. Khng c qu 3,5%. Khng pht hin c. Khng c qu 2,0 mg/kg.

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CNG BO/S 539 + 540/Ngy 27-10-2011

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

CNG BO/S 539 + 540/Ngy 27-10-2011

13

Ph lc 15 YU CU K THUT V PHNG PHP TH I VI GM GELLAN 1. Tn khc, ch s 2. nh ngha INS 418 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 ha 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 Phi c phn ng to gel vi ion calci c trng. calci To gel vi ion natri Phi c phn ng to gel vi ion natri c trng. 5.2. tinh khit Gim khi lng Khng c qu 15,0% (nhit sy 1050C trong 2,5 gi). khi sy kh Nitrogen Khng c qu 3,0% Isopropyl alcol Khng c qu 750,0 mg/kg (m t phn Phng php th) Ch Khng c qu 2,0 mg/kg. 5.3. Cc yu cu v vi sinh vt Tng s vi sinh vt Khng c qu 10.000 khun lc/g

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CNG BO/S 539 + 540/Ngy 27-10-2011

E. coli Salmonella Nm men v nm mc 5.4. Hm lng

m tnh i vi mu th m tnh i vi mu th Khng c qu 400 khun lc/g 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 Cho 1,0 g mu vo 99 ml nc v khuy trong 2 gi, s calci 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 Cho 1,0 g mu vo 99 ml nc v khuy trong 2 gi, s natri 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 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 Dow-Corning 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

CNG BO/S 539 + 540/Ngy 27-10-2011

15

Ch

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 ha 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) - 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. Cc yu cu v 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 ha tan hon ton. Gii hn thi gian ha 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.

16

CNG BO/S 539 + 540/Ngy 27-10-2011

E.coli

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. S dng k thut v trng, ha 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 ha 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

CNG BO/S 539 + 540/Ngy 27-10-2011

17

Salmonella

Nm men v nm mc

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 ha 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. S dng k thut v trng, phn tn 1 g mu trong 99 ml dung dch m phosphat v dng Stomacher, my lc hoc my khuy cho n khi tan hon ton. Gii hn thi gian ha 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 ha 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. 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.

6.4. nh lng

18

CNG BO/S 539 + 540/Ngy 27-10-2011

Ph lc 16 YU CU K THUT V PHNG PHP TH I VI PECTIN 1. Tn khc, ch s INS 440 2. nh ngha Pectin bao gm ch yu l cc ester ca acid polygalacturonic c methyl ha 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 ha. 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 ha 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 ha 5. Yu cu k thut 5.1. nh tnh tan Pectin Nhm amid 5.2. tinh khit Gim khi lng khi sy kh SO2 D lng dung mi Tro khng tan trong acid Tan trong nc, khng tan trong methanol, ethanol v isopropanol Phi c phn ng c trng ca pectin. Ch p dng i vi cc pectin c amid ha (m t phn Phng php th) Khng c qu 12,0 % (sy 1050C trong 2 gi) Khng c qu 50 mg/kg. Khng vt qu 1% methanol, ethanol v 2-propanol dng n cht hoc hp cht Khng vt qu 1,0%

CNG BO/S 539 + 540/Ngy 27-10-2011

19

Tng s cht khng Khng qu 3,0%. tan 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 ha Khng vt qu 25,0% tng s cc nhm carboxyl ca pectin. Ch Khng c qu 5,0 mg/kg. 6. Phng php th 6.1. nh tnh Lm m 0,05 g mu bng 2 - propanol. B sung 50 ml nc Pectin 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 dch m Dung dch pH = 7,0 *) mu th Nc Dung dch enzym **)

Khng c 0,5 ml 1,0 ml 1,0 ml enzym Khng c 0,5 ml 0,5 ml 1,5ml mu th Mu th 0,5 ml 1,0 ml 0,5ml 0,5 ml *) Ha 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 ha 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.

20

CNG BO/S 539 + 540/Ngy 27-10-2011

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 thy tinh xp ra 6 ln mi ln 10 ml hn hp HCl-ethanol 60%. ha 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 thy 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 300C. 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. 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 thy tinh kt ni t sinh hn ti bnh thu hoc ng ch U cha 10 ml dung dch hydrogen peroxyd 3% c trung ha 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. 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.

6.2. tinh khit SO2

D lng dung mi

CNG BO/S 539 + 540/Ngy 27-10-2011

21

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 ha 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 Wchun 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. Tng s cht khng Sy giy lc si thy tinh 70 mm (GF/B (Whatman code tan 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 tetra-acetic 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).

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CNG BO/S 539 + 540/Ngy 27-10-2011

Acid galacturonic v mc amid ha

Lc chn khng dung dch nng qua giy lc si thy 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 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 thy 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 ha (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

CNG BO/S 539 + 540/Ngy 27-10-2011

23

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. ha 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 ha 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. 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 ha (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 ha, 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 ha.

24

CNG BO/S 539 + 540/Ngy 27-10-2011

Ch 3: Nu mun, tnh ester ha (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

- 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

25

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 gi tr khong 2000) Dng ht mn, si nh, hoc bt, mu trng hoc trng nht, khng c mi, ht m. Cht nh ha, cht n nh, cht lm dy.

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh

26

CNG BO/S 539 + 540/Ngy 27-10-2011

tan

To bt To kt ta 5.2. tinh khit Gim khi lng khi lm kh pH Tro sulfat

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. Phi c phn ng to bt c trng. Phi c phn ng to kt ta c trng. Khng c qu 10,0% (sy ti 105C, trong 3 gi).

5,0 - 8,0 (dung dch 1/100). Khng c qu 1,5 %. (Th theo hng dn ti JECFA monograph 1 - Vol.4 - Phng php I, cn 1 g mu). Ch Khng c qu 2,0 mg/kg. 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. 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. - 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. Xc nh hm lng nhm methoxyl bng phng php Xc nh nhm Ethoxyl v Methoxyl (xem JECFA monograph 1 - Vol.4 )

To kt ta

6.2. tinh khit Ch

6.3. nh lng

CNG BO/S 539 + 540/Ngy 27-10-2011

27

Ph lc 18 YU CU K THUT V PHNG PHP TH 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

28

CNG BO/S 539 + 540/Ngy 27-10-2011

tan To bt To kt ta Mc thay th 5.2. tinh khit Gim khi lng khi lm kh Tro sulfat Ch 5.3. Hm lng

Trng n trong nc, to dung dch t trong n trng c, nht, keo; khng tan trong ethanol. Phi c phn ng to bt c trng. Phi c phn ng to kt ta c trng. Xc nh mc thay th bng sc k kh.

Khng c qu 15% i vi dng que, khng c qu 10% i vi dng bt (sy n khi lng khng i). Khng c qu 0,6 %. Khng c qu 2,0 mg/kg. 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). 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 - Th theo hng dn ti JECFA monograph 1 - Vol.4 Tro sulfat Phng php I. - Cn 1 g mu. 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 Xc nh nhm ethoxyl (c th xc nh ring bit ethoxyl v methoxyl bng sc k kh (Cobler, Samsel and Beaver,

CNG BO/S 539 + 540/Ngy 27-10-2011

29

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 mt ng sc kh chy t c ti y bnh dn hi 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 155C 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 105C 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 155C 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,

30

CNG BO/S 539 + 540/Ngy 27-10-2011

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 dch 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 quy 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 qu 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 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

31

Ph lc 19 YU CU K THUT V PHNG PHP TH 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

32

CNG BO/S 539 + 540/Ngy 27-10-2011

tan To bt To kt ta Phn ng mu 5.2. tinh khit Gim khi lng khi lm kh pH Natri Natri clorid Glycolat t do thay th Ch 5.3. Hm lng

To dung dch keo nht trong nc, khng tan trong ethanol. Phi c phn ng to bt c trng. Phi c phn ng to kt ta c trng. Phi c phn ng mu c trng. Khng c qu 12,0 % sau khi sy (sy ti 105C n khi lng khng i) 6,0 8,5 (dung dch 1 trong 100). Khng c qu 12,4 % tnh theo ch phm lm kh. Khng c qu 0,5 % tnh theo ch phm lm kh (m t trong phn Phng php th). Khng c qu 0,4 % tnh theo natri glycolat tnh theo ch phm lm kh (m t trong phn Phng php th). Khng c nh hn 0,2 v khng c qu 1,5 (m t trong phn Phng php th). Khng c qu 2,0 mg/kg. 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

33

6.2. tinh khit Natri clorid

Glycolat t do

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 mu trong l in 15 pht nhit 500C. 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). 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:

34

CNG BO/S 539 + 540/Ngy 27-10-2011

thay th

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 quy 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). 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 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 110C 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 quy 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

CNG BO/S 539 + 540/Ngy 27-10-2011

35

Ch

6.3. nh lng

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 quy 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)] - 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. 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).

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CNG BO/S 539 + 540/Ngy 27-10-2011

Ph lc 20 YU CU K THUT V PHNG PHP TH 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 ta Phi c phn ng to kt ta c trng. To c Phi c phn ng to c c trng. Gii phng Khi un nng mu th vi soda vi, kh amoniac s c amoniac gii phng. 5.2. tinh khit Gim khi lng Khng c qu 18% (sy ti 100 - 105o trong 6 gi). khi lm kh Cc hp cht khng Dung dch mu th 1/40 nng khng c c mi kh chu. tan trong nc v Khi quan st lp dy 2 cm, hu nh trong, ch c php trng c nh sa rt nh. c mi Lu hunh dioxyd Khng c qu 40,0 mg/kg.

CNG BO/S 539 + 540/Ngy 27-10-2011

37

Tro Khng c qu 2,0%. Arsen Khng c qu 1,0 mg/kg. Ch Khng c qu 1,5 mg/kg. Cadimi Khng c qu 0,5 mg/kg. Thy ngn Khng c qu 0,15 mg/kg. 5.3. Cc yu cu v vi sinh vt Tng s vi sinh vt < 104 /g hiu kh Enterobacteriaceae < 10/g hoc vi khun nhm coli-aerogens Streptococci nhm < 102 / g Lancefield D 6. Phng php th 6.1. nh tnh To kt ta 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 ta 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 - 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. Cadimi - Th theo hng dn ti JECFA monograph 1 - Vol.4. - Phng php II. Thy ngn - Th theo hng dn ti JECFA monograph 1 - Vol.4. - Phng php II.

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CNG BO/S 539 + 540/Ngy 27-10-2011

QCVN 4 - 22: 2011/BYT

QUY CHUN K THUT QUC GIA V PH GIA THC PHM - CHT NH HA National technical regulation on Food Additive - Emulsifier

Li ni u QCVN 4-22: 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

39

QUY CHUN K THUT QUC GIA V PH GIA THC PHM - CHT NH HA National technical regulation on Food Additive - Emulsifier 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 nh ha 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 nh ha 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 nh ha: l ph gia thc phm c b sung vo thc phm nhm mc ch to ra hoc duy tr dng nh tng ng nht ca hai hay nhiu thnh phn 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. MTDI (Maximum tolerable daily intake): Lng n vo hng ngy ti a chu ng c. 3.7. INS (International numbering system): H thng m s quc t v ph gia thc phm. II. YU CU K THUT, PHNG PHP TH V LY MU 1. Yu cu k thut v phng php th i vi cc cht nh ha c quy nh ti cc ph lc ban hnh km theo Quy chun ny nh sau:

40

CNG BO/S 539 + 540/Ngy 27-10-2011

1.1. 1.2. 1.3. 1.4. 1.5. 1.6. 1.7. 1.8. 1.9.

Ph lc 1: Ph lc 2: Ph lc 3: Ph lc 4: Ph lc 5: Ph lc 6: Ph lc 7: Ph lc 8: Ph lc 9:

1.10. Ph lc 10: 1.11. Ph lc 11: 1.12. Ph lc 12: 1.13. Ph lc 13: 1.14. Ph lc 14: 1.15. Ph lc 15: 1.16. Ph lc 16: 1.17. Ph lc 17: 1.18. Ph lc 18: 1.19. Ph lc 19:

Yu cu k thut v phng php th i vi monov di-glycerid ca acid bo Yu cu k thut v phng php th i vi ester ca glycerol vi acid lactic v acid bo Yu cu k thut v phng php th i vi ester ca glycerol vi acid citric v acid bo Yu cu k thut v phng php th i vi ester ca glycerol vi acid diacetyl tartric v acid bo Yu cu k thut v phng php th i vi sucroglycerid Yu cu k thut v phng php th i vi ester ca polyglycerol vi cc acid bo Yu cu k thut v phng php th i vi stearyl citrat Yu cu k thut v phng php th i vi trikali ortho-phosphat Yu cu k thut v phng php th i vi cc mui amoni ca acid phosphatidic Yu cu k thut v phng php th i vi sucrose acetat isobutyrat Yu cu k thut v phng php th i vi ester glycerol ca nha cy Yu cu k thut v phng php th i vi dinatri diphosphat Yu cu k thut v phng php th i vi calci polyphosphat Yu cu k thut v phng php th i vi cc mui ca acid myristic, palmitic v stearic (Ca, Na, K, NH4) Yu cu k thut v phng php th i vi ester ca sucrose vi cc acid bo Yu cu k thut v phng php th i vi dioctyl natri sulphosucinat Yu cu k thut v phng php th i vi stearyl tartrat Yu cu k thut v phng php th i vi sorbitan monostearat Yu cu k thut v phng php th i vi sorbitan tristearat

CNG BO/S 539 + 540/Ngy 27-10-2011

41

1.20. Ph lc 20: 1.21. Ph lc 21: 1.22. Ph lc 22:

Yu cu k thut v phng php th i vi sorbitan monolaurat Yu cu k thut v phng php th i vi sorbitan monooleat Yu cu k thut v phng php th i vi sorbitan monopalmitat

2. Cc yu cu k thut quy nh trong Quy chun ny c th theo JECFA monograph 1 - Vol. 4, ngoi tr mt s php th ring c m t trong cc ph lc. Cc phng php th c hng dn trong Quy chun ny khng bt buc phi p dng, c th s dng cc phng php th khc tng ng. 3. Ly mu theo hng dn ti Thng t s 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 nh ha 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 nh ha Vic kim tra cht lng, v sinh an ton i vi cc cht nh ha 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 nh ha sau khi hon tt ng k bn cng b hp quy v bo m cht lng, v sinh an ton, ghi nhn ph hp vi cc quy nh ca php lut. V. T CHC THC HIN 1. Giao Cc An ton v sinh thc phm ch tr, phi hp vi cc c quan chc nng c lin quan hng dn trin khai v t chc vic thc hin Quy chun ny. 2. Cn c vo yu cu qun l, Cc An ton v sinh thc phm c trch nhim kin ngh B Y t sa i, b sung Quy chun ny. 3. Trng hp hng dn ca quc t v phng php th v cc quy nh ca php lut vin dn trong Quy chun ny c sa i, b sung hoc thay th th p dng theo vn bn mi.

42

CNG BO/S 539 + 540/Ngy 27-10-2011

Ph lc 1 YU CU K THUT V PHNG PHP TH I VI MONO V DIGLYCERID CA CC ACID BO 1. Tn khc, ch s Glyceryl monostearate, glyceryl monopalmitate, glyceryl monooleate; monostearin, monopalmitin, monoolein; GMS (i vi glyceryl monostearate); INS 471 Hn hp cc ester mono- v diglyceryl mch di vi cc acid bo bo ha v khng bo ha c trong cht bo thc phm; c cha hm lng alpha-monoglycerid khng thp hn 30% v cng c th c cha cc monoglycerid ng phn; cng nh di- v triglycerid, glycerol t do, cc acid bo t do, sn phm x phng ha v nc; thng c sn xut bng cch thy phn tch glyceryl ca cc cht bo v du thc phm, nhng cng c th c sn xut bng cch este ha cc acid bo vi glycerol c hoc khng qua chng ct phn t.

2. nh ngha

Cng thc cu to

Trong -OCR l ca acid bo Khi lng phn t Glyceryl monostearat: 358,6 Glyceryl distearat: 625,0 3. Cm quan Cht bo rn dng sp c mu trng hoc kem hoc dng lng snh 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan Khng tan trong nc, tan trong ethanol, cloroform v benzen. Hp th hng Phi c ph hng ngoi c trng ca ester mt phn ca ngoi acid bo vi polyol. Acid bo Phi c phn ng c trng ca acid bo. Glycerol Phi c phn ng c trng ca glycerol. 5.2. tinh khit Nc Khng c qu 2,0 % (phng php Karl Fischer)

CNG BO/S 539 + 540/Ngy 27-10-2011

43

Ch s acid Khng c qu 6. Glycerol t do Khng c qu 7% X phng Khng c qu 6%, tnh theo natri oleat. Ch Khng c qu 2,0 mg/kg. 6. Phng php th tinh khit X phng Thm 10 g mu th vo hn hp gm 60 ml aceton v 0,15 ml dung dch xanh bromophenol (0,5%) c trung ha trc bng acid hydrocloric 0,1N hoc natri hydroxyd 0,1N. Lm m t t trn cch thy cho n khi dung dch tan hon ton, chun bng acid hydrocloric 0,1N cho n khi mu xanh bin mt. yn dung dch trong 20 pht, lm m cho n khi cn c ha tan v mu xanh ti xut hin, tip tc chun n mt mu xanh. Mi ml acid hydrocloric 0,1N tng ng vi 0,0304 g C18H33O2Na. 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.

44

CNG BO/S 539 + 540/Ngy 27-10-2011

Ph lc 2 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI ACID LACTIC V ACID BO 1. Tn khc, ch s 2. nh ngha Lactic acid esters of mono- and diglycerides; lactoglycerides INS 472b L hn hp cc ester ca glycerol vi acid lactic v acid bo trong cht bo thc phm. Sn phm thng mi c th ch r hm lng monglycerid, acid lactic, ch s acid, ch s x phng ha, hm lng acid bo t do, im ng c ca cc acid bo t do, ch s iod, hm lng glycerol t do v hm lng nc.

Cng thc cu to

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Khng tan trong nc lnh nhng c th phn tn trong nc nng. Acid bo Phi c phn ng c trng ca acid bo. Acid lactic Phi c phn ng c trng ca acid lactic. Glycerol Phi c phn ng c trng ca glycerol. 5.2. tinh khit Cc acid Ch c acid bo v acid lactic. Ch Khng c qu 2,0 mg/kg. 6. Phng php th tinh khit 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.

Trong R1, R2 v R3 l ca acid bo, acid lactic hoc hydrogen (theo thnh phn) Cht rn dng sp Cht nh ha

CNG BO/S 539 + 540/Ngy 27-10-2011

45

Ph lc 3 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI ACID CITRIC V ACID BO 1. Tn khc, ch s Citric acid esters of mono- and di-glycerides, citroglycerides, CITREM; INS 472c Thu c nh qu trnh ester ha glycerol vi acid citric v acid bo thc phm hoc nh phn ng ca hn hp mono v diglycerid ca acid bo vi acid citric; ch phm bao gm cc ester hn hp ca acid citric v cc acid bo vi glycerol; c th cha lng nh cc acid bo t do, glycerol t do, acid citric t do v mono - v diglycerid; c th c trung ha hon ton hoc mt phn bng NaOH hoc KOH (nh c cng b trn nhn mc).

2. nh ngha

Cng thc cu to

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Acid bo Acid citric Glycerol 5.2. tinh khit Tro sulfat

Trong : t nht mt trong R1, R2 hoc R3 l ca acid citric, mt l ca acid bo v R cn li c th l ca acid citric, acid bo hoc hydrogen. Dng du ti sp, mu trng n trng ng Cht n nh, cht nh ha, cht chng oxy ha

Khng tan trong nc lnh, c th phn tn trong nc nng; tan trong du v m; khng tan trong ethanol lnh Phi c phn ng c trng ca acid bo . Phi c phn ng c trng ca acid citric. Phi c phn ng c trng ca glycerol. Sn phm khng c trung ha: Khng c qu 0,5% Sn phm c trung ha hon ton hoc mt phn: Khng c qu 10% (S dng 2 g mu th, phng php I) Khng c qu 4% 8 - 33% (m t trong phn Phng php th)

Glycerol t do Glycerol tng s

46

CNG BO/S 539 + 540/Ngy 27-10-2011

Acid citric tng s Acid bo tng s Ch 6. Phng php th tinh khit Glycerol tng s

13 - 50% (m t trong phn Phng php th) 37 - 81% (m t trong phn Phng php th) Khng c qu 2,0 mg/kg.

Cn chnh xc 2 g mu v cho vo bnh x phng ha, thm vo 50 ml dung dch KOH 0,5 M trong ethanol, v un si vi sinh hn ngc trong 30 pht. Thm chnh xc 99 ml (t buret) chloroform v 25 ml acid acetic bng vo bnh nh mc dung tch 1 lt. Chuyn ton lng dch trong bnh x phng ha vo bnh nh mc dung tch 1 lt, cho 3 ln 25 ml nc. Thm vo 500 ml nc v lc mnh trong 1 pht. Pha n th tch 1 lt bng nc, lc u v yn cho tch thnh cc lp. Dng pipet ly 50 ml dung dch acid acetic periodic TS vo trong cc cc dung tch 400 ml. Chun b 2 mu trng bng cch thm 50 ml nc vo mi cc. Dng pipet ly 50 ml dung dch mu trong nc cho vo mt trong cc cc cha 50 ml acid acetic periodic TS, lc nh cho u, y bng tm knh nhn qua c v yn trong 30 pht. Nhng khng c qu 1,5 gi. Thm 20 ml dung dch KI 15%, lc nh cho u v yn t nht 1 pht nhng khng c qu 5 pht. Khng c di nh sng trc tip hoc nh sng trng. B sung 100 ml nc v chun bng dung dch natri thiosulfat 0,1 N. S dng dng my khuy lm u dung dch. Tip tc chun cho n khi mt mu iod nu trong dung dch nc. Thm 2 ml tinh bt TS v tip tc chun cho n khi mt mu iod trong lp chloroform c tch ra trong qu trnh chun v mt mu xanh ca tinh bt iodo trong dung dch nc. Kt qu: Glycerol tng s (%): Trong : B: Lng dung dch natri thiosulfat dng chun mu trng cha 50 ml nc S: Lng dung dch natri thiosulfat dng chun mu th N: Nng dung dch natri thiosulfat 0,1 N

CNG BO/S 539 + 540/Ngy 27-10-2011

47

W: Khi lng mu th c ly bng pipet em chun v c tnh theo cng thc: Trong : a: Khi lng mu (g) em x phng ha Acid citric tng s Nguyn tc: Mu c x phng ha bng dung dch KOH trong alcol v cc acid bo c tch ra bng trch ly. Acid citric c chuyn thnh cc dn sut trimethylsilyl (TMS) v c phn tch bng sc k kh lng. X phng ha: Cn chnh xc 1 g mu cho vo bnh y trn, thm 25 ml dung dch KOH 0,5M trong ethanol v un si vi sinh hn ngc trong 30 pht. Acid ha hn hp bng HCl v cho bc hi bng my c quay hoc bng phng php thch hp khc. Trch ly: Chuyn ton lng mu trong bnh vo mt bnh gn, s dng khng qu 50 ml nc v trch ly bng 3 ln 50 ml heptan, loi b phn dch chit heptan tch ra. Chuyn lp dung dch nc vo bnh nh mc dung tch 100 ml, trung ha pha ti th tch 100 ml bng nc v lc u. Dn xut ha: Dng pipet ly 1 ml dung dch pha c trn, 1 ml dung dch acid tartric (1 mg/ml trong nc) cho vo bnh y trn c th y np dung tch 10 ml v c n kh. Thm 1 ml pyridin, 0,2 ml dung dch trimethyl-chlorosilan (TMCS), 0,4 ml hexamethyl-disilazan (HMDS), 0,1 ml n-methyl-n-trimethylsilyl-trifluoroacetamid (MSTFA). y np cht v lc cn thn c dung dch tan hon ton. Gia nhit bnh trong t sy 600C trong 1 gi. Sc k kh: Bt k sc k kh thch hp c th c dng vi detector ion ha ngn la v ct (thy tinh, chiu di 1,8 m v ng knh trong 2 mm), c nhi bng DC-200 10% trn chromosorb Q (80/100 mesh). Cc iu kin cho sc k kh: nhit l ct 1650C, nhit bung tim 2400C, nhit detector 2400C; tc dng kh mang nitrogen 24 ml/pht.

48

CNG BO/S 539 + 540/Ngy 27-10-2011

Cch tin hnh: Bm 5 l mu cc dn xut TMS. Thi gian lu i vi acid tartric khong 12 pht v thi gian lu tng i ca acid citric/acid tartric khong 2,3. Lp li cch tin hnh c m t trn trong phn Dn xut ha v Sc k kh s dng 1 ml dung dch i chng acid citric (3 mg/ml trong nc) thay th 1 ml dung dch mu th. Tnh kt qu: o din tch mi pic bng phng php thch hp. Acid citric tng (%) = Trong : ACS: Din tch pic ca acid citric (dung dch mu) ATS: Din tch pic ca acid tartric (dung dch mu) ATR: Din tch pic ca acid tartric (dung dch i chng) ACR: Din tch pic ca acid citric (dung dch i chng) WCR: Khi lng (g) ca acid citric trong 1 ml dung dch i chng W: Khi lng (g) ca mu ester glycerol vi acid citric v acid bo. Cn chnh xc 5 g mu cho vo bnh y trn dung tch 250 ml, thm vo 50 ml dung dch KOH 1 N trong ethanol, v un si vi ng sinh hn ngc trn ni cch thy trong 1 gi. Chuyn ton lng dung dch trong bnh x phng ha vo phu chit dung tch 1.000 ml, cho 3 ln 25 ml nc v thm 5 git da cam methyl TS. Thm cn thn dung dch HCl m c cho n khi mu ca dung dch chuyn sang mu r rt v lc u tch cc acid bo. Trch ly cc acid bo tch c bng ba ln 100 ml dung mi diethyl ether. Ha chung cc phn tch c v ra mi ln bng 50 ml dung dch NaCl 10% cho n khi dung dch NaCl c ra tr nn trung tnh. Lm kh dung dch ether bng Na2SO4 khan. Sau c bc hi trn cch thy ether, thm 10 pht na trn cch thy v cn phn cn.

Acid bo tng s

CNG BO/S 539 + 540/Ngy 27-10-2011

49

Ph lc 4 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI ACID DIACETYL TARTRIC V ACID BO 1. Tn khc, ch s Diacetyltartric acid esters of mono- and diglycerides, DATEM; INS 472e Tartric, acetic and fatty acid esters of glycerol, mixed; Mixed acetic and tartric acid esters of mono and diglycerides of fatty acids; INS 472f Sn phm gm hn hp cc ester ca glycerol vi acid mono, diacetyltartric v cc acid bo thc phm. N c tng hp bng phn ng gia anhydrid diacetyltartric v mono, diglycerid ca acid bo c mt acid acetic, hoc bng cch tng tc gia anhydrid acetic v mono, diglycerid ca acid bo c mt acid tartric. Do trao i nhm acyl trong ni phn t hoc gia cc phn t, c hai phng php sn xut u to ra cc thnh phn chnh nh nhau, vic to ra cc thnh phn chnh ph thuc vo t l tng i ca cc nguyn liu c bn, vo nhit v thi gian phn ng. Sn phm ny c th cha mt lng nh glycerol t do, cc acid bo t do, acid tartric v acid acetic t do. Sn phm thng mi c th c quy nh cc ch tiu k thut r hn nh ch s acid, hm lng acid tartric tng s, hm lng acid acetic t do, ch s x phng ha, ch s iod, hm lng acid bo t do, im ng c ca acid bo t do. Cc thnh phn chnh l:

2. nh ngha

Cng thc cu to

Trong : 1) Mt hoc hai nhm R l ca acid bo 2) Nhm R khc l ca hoc - acid diacetylat tartric - acid monoacetylat tartric - acid tartric - acid acetic - hydrogen 3. Cm quan T dng lng, bt nho n dng rn ging sp (vy hoc bt) 4. Chc nng Cht nh ha 5. Yu cu k thut

50

CNG BO/S 539 + 540/Ngy 27-10-2011

5.1. nh tnh tan

1, 2- diol Acid bo Acid acetic Acid tartric Glycerol 5.2. tinh khit Acid Khng pht hin c cc loi acid khc ngoi acid acetic, acid tartric v acid bo. Tro sulfat Khng c qu 0,5% c xc nh 800 25C Acid bo t do Khng c qu 3% tnh theo acid oleic Acid acetic tng Khng c nh hn 8% v khng c qu 32% sau khi thy s phn (m t trong phn Phng php th) Acid tartric tng Khng c nh hn 10% v khng c qu 40% sau khi x s phng ha (m t trong phn Phng php th) Glycerol tng s Khng c nh hn 11% v khng c qu 28% sau khi x phng ha (m t trong phn Phng php th) Glycerol t do Khng c qu 2,0% Ch Khng c qu 2,0 mg/kg. 6. Phng php th 6.1. nh tnh 1, 2- diol Cho vi git ch acetat TS vo dung dch c 500 mg mu trong 10 ml methanol. Cht kt ta vn, mu trng khng tan c hnh thnh. 6.2. tinh khit Acid acetic tng Dng c: s Lp rp b dng c chng ct Hortvet - Sellier ci tin nh hnh v sau, s dng ng Sellier c kch thc bn trong ln (khong 38-x 203 mm) v by chng ct dung tch ln. Cch tin hnh: Cn chnh xc 4 g mu v cho vo ng bn trong ca b dng c v lp ng vo bnh ngoi c cha khong 300 ml nc nng va un si. Thm 10 ml acid perchloric 4 N [35 ml (60 g) acid perchloric 70% trong 100 ml nc] vo mu th, v ni ng bn trong vi sinh hn lm mt bng nc qua by chng ct. Chng ct bng cch gia nhit bnh pha ngoi sao cho thu c 100 ml dung dch ct trong khong 20 - 25 pht. Tp hp cc phn dch ct c 100 ml v thm phenolphthalein TS vo mi phn, chun bng dung dch NaOH 0,5 N. Tip tc chng ct cho n khi phn 100 ml dung dch ct c cn

Phn tn trong nc nng v nc lnh, tan trong methanol v ethanol. Phi c phn ng c trng ca 1,2-diol. Phi c phn ng c trng ca acid bo Phi c phn ng c trng ca acid acetic Phi c phn ng c trng ca acid tartric Phi c phn ng c trng ca glycerol

CNG BO/S 539 + 540/Ngy 27-10-2011

51

Acid tartric tng s

khng qu 0,5 ml dung dch NaOH 0,5N trung ha. (Ch : khng c chng ct n cn kh). Tnh khi lng (mg) acid bay hi trong mu theo cng thc V x e, trong V l tng th tch (ml) ca dung dch NaOH 0,5N dng chun v e l h s ng lng 30,03. ng chun: Chuyn 100 mg acid tartric tinh khit c cn chnh xc vo bnh nh mc dung tch 100 ml, ha tan acid tartric trong 90 ml nc v thm nc n th tch 100 ml, lc u. Ly cc phn 3, 4, 5 v 6 ml cho vo cc cuvet ring bit ph hp (kch thc 19 x 150 mm), thm lng nc thch hp c dung dch th tch 10 ml. Cho vo mi cuvet 4 ml dung dch natri metavanadat 5% va mi pha, 1 ml acid acetic. (Ch : s dng cc dung dch ny trong vng 10 pht sau khi mu tng ln). Chun b mu trng ging nh cch trn, s dng 10 ml nc thay th dung dch acid tartric. t thit b v tr bng 0 i vi mu trng v sau o hp th ca 4 dung dch acid tartric ti bc sng 520 nm vi my quang ph thch hp hoc my so mu quang in c trang b knh lc 520 nm. T s liu thu c, xy dng ng chun trn h trc to vung gc vi trc tung biu din hp th, trc honh biu din lng acid tartric tng ng (mg). Chun b mu th: Cn chnh xc 4 g mu cho vo bnh Erlenmeyer dung tch 250 ml, thm vo 80 ml dung dch KOH 0,5N v 0,5 ml phenolphthalein TS. Ni bnh vi mt sinh hn khng kh c chiu di t nht 65 cm, gia nhit hn hp trn tm nhit trong 2,5 gi. Thm vo hn hp nng khong 10% acid phosphoric cho n khi hn hp c phn ng acid r vi giy ch th Congo. Ni li bnh vi sinh hn khng kh v gia nhit cho n khi acid bo c ha lng v trong. Lm ngui v sau chuyn hn hp vo mt bnh gn dung tch 250 ml vi s h tr ca cc phn nh nc v chloroform. Chit cc acid bo c gii phng ra bng 3 ln 25 ml nc lin tip v cho phn nc ra vo bnh gn c cha lp nc. Chuyn ton b hn hp trong bnh gn u vo cc dung tch 250 ml, gia nhit trn ni cch thy loi b cc vt ca chloroform, lc qua giy lc mn c ra acid vo bnh nh mc dung tch 500 ml, v cui cng pha ti th tch 500 ml bng nc (Dung dch I). Dng pipet ly 25 ml dung dch ny cho vo bnh nh mc dung tch 100 ml v pha ti th tch 100 ml bng nc (Dung dch II). Gi phn cn li ca Dung dch I xc nh glycerol tng s. Cch tin hnh: Cho 10 ml dung dch II c chun b trong phn Chun b

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CNG BO/S 539 + 540/Ngy 27-10-2011

Glycerol tng s

mu th vo cuvet 19 x 150 mm v tip tc lm theo hng dn trong phn ng chun, bt u vi cho vo mi cuvet 4 ml dung dch metavanadat 5%. T ng chun xc nh khi lng (mg) acid tartric trong pha long cui cng, sau nhn vi 20 v chia kt qu cho khi lng mu gc thu c % acid tartric. Chuyn 5 ml dung dch I chun b trong phn xc nh Acid tartric tng s vo trong bnh Erlenmeyer c np thy tinh dung tch 250 ml hoc bnh iod. Thm vo bnh 15 ml acid acetic bng v 25 ml dung dch acid periodic, c chun b bng cch ha tan 2,7 g acid periodic (H5IO6) trong 50 ml nc, thm 950 ml acid acetic bng v lc mnh; bo qun dung dch ny trnh nh sng. Lc u hn hp trong 1 hoc 2 pht, yn trong 15 pht, thm 15 ml dung dch KI 15% v 15 ml nc, lc xoy, yn trong 1 pht v sau chun iod c gii phng ra bng dung dch natri thiosulfat 0,1N, dng tinh bt TS l cht ch th. Thc hin chun mu trng s dng nc thay th mu th. Th tch thc l s ml dung dch natri thiosulfat 0,1N dng chun glycerol v acid tartric trong mu th c 5 ml Dung dch I. T % xc nh c trong Phng php th Acid tartric tnh th tch dung dch natri thiosulfat 0,1N dng chun acid tartric. Chnh lch gia th tch thc v th tch tnh ton c dng i vi acid tartric l s ml ca dung dch natri thiosulfat 0,1N tiu th bi glycerol trong mu. Mt ml dung dch natri thiosulfat 0,1N tng ng vi 2,303 mg glycerol v 7,505 mg acid tartric.

Dng c chng ct Hortvet-Sellier ci tin 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.

CNG BO/S 539 + 540/Ngy 27-10-2011

53

Ph lc 5 YU CU K THUT V PHNG PHP TH I VI SUCROGLYCERID 1. Tn khc, ch s 2. nh ngha INS 474

Sucroglycerid thu c nh phn ng gia sucrose vi cht bo hoc du n c hoc khng c mt ca dung mi. Sn phm gm c hn hp mono- v di-ester ca sucrose v acid bo kt hp vi mono-, di- v triglycerid t cht bo hoc du. Ch cc dung mi sau y c th c s dng trong sn xut: dimethyl formamid, cyclohexan, isobutanol, isopropanol v ethyl acetat. 3. Cm quan Khng mi, dng mm, dng c, dng bt c mu trng n trng nht, hoc dng gel cng. 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan Khng tan trong nc lnh, tan trong ethanol. Acid bo Phi c phn ng c trng ca acid bo. ng Phi c phn ng c trng ca ng. 5.2. tinh khit Tro sulfat Khng c qu 2%. Th 2 g mu (Phng php I) Ch s acid Khng c qu 6. Sucrose t do Khng c qu 5% (m t trong phn Phng php th) DimethylKhng c qu 1 mg/kg (m t trong phn Phng php th) formamid Cyclohexan v Khng c qu 10 mg/kg, dng n cht hoc hp cht (m isobutanol t trong phn Phng php th) Ethyl acetat v Khng c qu 350 mg/kg, dng n cht hoc hp cht isopropanol Ch 5.3. Hm lng Khng c qu 2,0 mg/kg. Khng c nh hn 40% v khng c ln hn 60% ester sucrose

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CNG BO/S 539 + 540/Ngy 27-10-2011

6. Phng php th 6.1. nh tnh Acid bo Cho 1 ml ethanol vo 0,1 g mu, lm m v ha tan, thm 5 ml dung dch H2SO4 long TS, gia nhit trong cch thy trong 30 pht v lm ngui. Cht c mu trng vng hoc du c hnh thnh, v c ha tan khi thm 3 ml diethyl ether. ng Cho vo 2 ml lp dung dch nc tch ra t cht c hoc du trong php th acid bo, 1 ml anthron TS vo trong ng th, cho chy cn thn vo thnh ng th; b mt ranh gii gia hai lp chuyn thnh mu xanh da tri hoc xanh l cy.

6.2. tinh khit Sucrose t do Xc nh bng sc k kh lng (xem Quyn 4) Ha cht, thuc th: - Dung dch ni chun: Dung dch cholesterol trong chloroform 5 mg/ml hoc tetracosan trong chloroform 10 mg/ml - Pyridin (lm kh c qua ry phn t) - N, O- Bis-(trimethylsilyl)-acetamid (BSA) - Trimethylchlorosilan (TMCS) Cch tin hnh: Cn chnh xc 20 - 50 mg mu cho vo bnh silyl ha, thm 1 ml dung dch ni chun, 1 ml pyridin, 0,5 ml N, O-bis (trimethylsilyl) acetamid (BSA) v 0,5 ml trimethylchlorosilan (TMCS). y np bnh v gia nhit 700C trong 30 pht. Bm 1l vo thit b sc k kh lng. Cc iu kin sc k: Ct: - Chiu di 0,3 m - ng knh trong 4 mm - Cht liu: thy tinh - c nhi bng dexil Kh mang: nitrogen Tc dng: 40 ml/pht Detector: FID Chng trnh nhit : gi 1600C trong 1 pht sau tng n 160 - 3750C vi tc 150C/pht. o cc din tch pic ca sucrose v ni chun. H s p ng (RF) c tnh t qu trnh chy sc k kh lng vi cc dung dch chun sucrose cha ni chun.

CNG BO/S 539 + 540/Ngy 27-10-2011

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Kt qu: mg ni chun x din tch pic sucrose RF = ----------------------------------------------------Din tch pic ni chun x mg sucrose v mg ni chun x din tch pic sucrose x 100 % sucrose t do = -----------------------------------------------------RF x din tch ni chun x mg mu Xc nh bng sc k kh lng Ha cht, thuc th: - Dimethyl formamid - Dimethylamin hydrochlorid - Methanol - Ethanol - HCl - NaOH Dung dch chun: Chun b dung dch gc dimethylaminhydroclorid trong ethanol 4,47 mg/ml (tng ng vi 4,0 mg/ml dimethyl formamid) v chun b mt lot cc dung dch chun tng ng vi 4; 2 v 1 g/ml dimethylformamid bng cch pha long dung dch gc vi dung dch NaOH 0,1% trong ethanol. Chun b mu: B dng c thy phn c trnh by trong phn Ph lc. Cn chnh xc 40 g mu cho vo bnh y trn dung tch 1.000 ml. Thm 500 ml dung dch NaOH 5% trong methanol, v gn bnh vo b dng c thy phn. t bnh ct Erlenmeyer cha 10 ml dung dch HCl 1% trong methanol vo b dng c thy phn. Gia nhit bnh y trn v chng ct vi sinh hn ngc c nc lm mt trong 1 gi, sau ct ly 50 ml dch ct c th dng li. C dch ct c cho n kh trong cch thy si. Ha tan phn cn bng mt lng nh ethanol, thm 2,5 ml dung dch NaOH 5% trong ethanol v pha long n th tch 25 ml bng ethanol chun b mt dung dch mu. Cch tin hnh: Bm 2 l dung dch mu vo thit b sc k kh lng vi cc iu kin di y.

Dimethylformamid

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ng chun: Chun b ng chun bng cch bm 2 l cc dung dch chun vo thit b sc k kh. Cc iu kin sc k: Ct: - Di: 2m - ng knh trong: 2 mm - Cht liu: thy tinh - Ct nhi: 10% amin 220 v KOH 10% trn nn chromosorb W yu 80/100 c ra acid yu - Luyn ct: Gia nhit ti 1300C qua m vi tc dng nitrogen 5 ml/pht Kh mang: nitrogen Tc dng: 17 ml/pht Detector: FID Nhit : - Bung bm mu: 198 50C - Ct: 600C. Tnh kt qu: Nng dimethylformamid (CDFA) tnh theo cng thc sau:

Cyclohexan v isobutanol

Trong : C: Nng dimethylformamid pht hin c (g/ml) W: Khi lng mu (g) Xc nh bng sc k kh lng (xem Quyn 4) s dng cc iu kin sau: Ha cht, thuc th: - Dimethylformamid (tinh khit dng cho GLC) - Cyclohexan (dng cho quang ph UV) - Isobutanol (dng cho phn tch) Dung dch chun: Chun b dung dch gc cyclohexan v isobutanol trong dimethylformamid bng cch dng pipet ly 130 l cyclohexan v 125 l isobutanol cho vo dimethylformamid v lm y n th tch 10 ml.

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Isopropanol v ethyl acetat

T dung dch gc trn pha mt lot cc dung dch chun pha long c 5, 10 v 20 mg/kg cyclohexan v isobutanol. V ng p ng bng cch bm 5 l cc dung dch chun pha long vo thit b sc k kh vi cc iu kin sau. Chun b mu th: Cn 5 g mu chnh xc n 10 mg cho vo bnh c np y thy tinh, thm vo 5 g dimethylformamid v lm m ha tan. Lm ngui v bm 5 l vo thit b sc k kh vi cc iu kin sau. Ct: - Di: 3m - ng knh trong: 4,5 mm - Cht liu: thp khng r - Cht nhi: 20% carbowax 20 M trn Chromosorb G 60/80 Kh mang: heli (p sut 1,6 bar) Detector: ion ha ngn la Nhit : - Bung bm mu: 1300C - Ct: 1300C - Detector: 2000C Xc nh nng cyclohexan v isobutanol trong dung dch mu (50%) bng cch so snh vi cc dung dch chun v nhn nng vi 2 chuyn cc kt qu cho ph hp vi sucroglycerid gc. Xc nh bng sc k kh (xem Quyn 4) c bung ly mu khng gian hi s dng cc iu kin sau: Ha cht, thuc th: - Isopropanol - Ethyl acetat Dung dch chun: Ly 1 g mi loi isopropanol v ethyl acetat vo mt bnh nh mc v thm nc cho n khi t th tch 100 ml, v pha long dung dch ny c cc dung dch c nng 0,02 0,4 g/100 ml. Nu cn, chun b cc dung dch chun cha ti 7 g/100 ml isopropanol v ethyl acetat. Cch tin hnh: Cn 1 g chnh xc n 0,1 g bt mu cho vo l mu. Thm 5 l nc vo l mu v np kn nhanh bng septum. l mu vo

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thit b sc k kh t trc chng trnh v bt u phn tch theo cc iu kin cp di y. ng chun: Ly 1 g bt ester ca sucrose vi cc acid bo, khng cha dung mi hoc cc dung mi tn d bit, cho vo l mu, thm 5 l dung dch chun v y kn bng septum. l mu vo thit b sc k kh t trc chng trnh v bt u phn tch theo cc iu kin cp di y v v c ng chun ca mi dung mi. Ct: - Di: 30 m - ng knh trong: 0,53 mm - Cht liu: mao qun silica - Fim mng: 100% methyl polysiloxan - Luyn ct: Gia nhit ti 600C trong 2 - 3 gi vi tc kh nitrogen khong 10 ml/pht. Kh mang: nitrogen Tc dng: 5 ml/pht Detector: ion ha ngn la Nhit : - Bung bm mu: 1100C - Ct: 400C. - Detector: 1100C Bung khng gian hi: - Lng mu: 1 g 0,1 g + 5 l - Nhit gia nhit mu: 800C - Thi gian gia nhit mu: 40 pht - Nhit syringe: 850C - Th tch mu dng hi: 0,4 ml Kt qu: c tnh theo cng thc: Trong : Ci: Nng dung mi i (mg/kg); Ai: Din tch pic dung mi i (v.giy.);

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Cfi: H s chuyn i dung mi i ( dc ca ng chun) (g/v.giy). 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. 6.2. nh lng Xc nh bng sc k lng cao p (xem Quyn 4) s dng cc iu kin sau: Chun b mu: Cn chnh xc 250 mg mu cho vo bnh nh mc 50 ml. Pha ti th tch 50 ml bng tetrahydrofuran v lc u. Lc qua mng lc 0,5 m. Cch tin hnh: Bm 100 l mu vo thit b sc k lng cao p n nh trc. Cc iu kin sc k: Ct: Styren-divinylbenzen copolymer i vi sc k thm thu qua gel (TSK-GEL G2000 (Supelco) hoc tng ng) Pha ng: tetrahydrofuran dng cho HPLC ui kh Tc dng: 0,7 ml/pht Detector: RI (ch s khc x) Nhit : - Ct: 380C. - Detector: 380C Ghi sc k trong khong 90 pht. Tnh % ester sucrose trong mu: % ester sucrose = 100 A/T Trong : A: Tng cc din tch pic i vi 3 thnh phn chnh, mono-, div tri-ester, ra gii ln lt khong 65, 68 v 73 pht. T: Tng din tch tt c cc pic ra gii trong khong 90 pht Ph lc:

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B dng c thy phn: a: Sinh hn ngc b: Sinh hn c: Bnh y trn d: Chu nc e: Bnh Erlenmeyer

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Ph lc 6 YU CU K THUT V PHNG PHP TH I VI ESTER CA POLYGLYCEROL VI CC ACID BO 1. Tn khc, ch s 2. nh ngha Polyglycerol fatty acid esters, glyceryl fatty acid esters; INS 475 L cc ester mt phn hn hp c to thnh bng phn ng glycerol polymer ha vi m v du n, hoc vi acid bo; c th cha mt lng nh mono-, di- v triglyceride, glycerol t do v polyglycerol, acid bo t do v mui natri ca cc acid bo; polymer ha c th thay i v c ch ra bng mt con s (th d l tri-, 3), l s trung bnh nhm (gc) glycerol tnh cho 1 phn t polyglycerol. Mt polyglycerol nht nh bao gm mt s phn b cc mnh phn t c trng cho polymer ha danh nh ca n. Bng cch thay i t l cng nh bn cht cc cht bo hoc acid bo phn ng vi polyglycerol, ngi ta thu c mt nhm ln v a dng cc sn phm ester ny; Cc sn phm ester thng mi cn c c trng vi 1 s ch tiu khc nh ch s x phng ha, im ng c ca acid bo t do, ch s iod, ch s hydroxyl v hm lng tro.

Cng thc cu to Trong gi tr trung bnh ca n khong l 3 v mi R1, R2, R3 c th l ca acid bo hoc hydro. 3. Cm quan Dng lng nh du ti rt nht, c mu vng sng n h phch; cht do hoc nha c mu vng nht n mu nu; cht rn hoc cht sp c mu vng nht n mu nu. 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan T rt a nc ti rt a du, nhng l mt loi cht c xu hng phn tn trong nc v ha tan trong dung mi hu c v cc loi du. Acid bo Phi c phn ng c trng ca acid bo. Glycerol v Phi c phn ng c trng ca glycerol v polyglycerol. polyglycerol 5.2. tinh khit Acid Khng pht hin c acid no ngoi acid bo.

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Phn polyglycerol gm c khng nh hn 70% di-, tri-, tetraglycerol v phi c khng qu 10% polyglycerol bng hoc cao hn heptaglycerol. Ch Khng c qu 2,0 mg/kg. 6. Phng php th 6.1. nh tnh Glycerol v Chm 5 - 20 l lp dung dch nc thu c trong php th acid polyglycerol bo Php th nh tnh cc nhm chc dc theo cc chm i chng glycerol trn giy Whatman s 3 v trin khai sc k i xung trong 36 gi vi h dung mi isopropanol: nc (90: 10 ). Vt glycerol di chuyn c 40 cm v tip sau l cc polyglycerol c pht hin ln lt pha di sau glycerol khi giy c phun hoc permanganat trong aceton hoc bc nitrat trong amoniac. 6.2. tinh khit 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.

Polyglycerol

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Ph lc 7 YU CU K THUT V PHNG PHP TH I VI STEARYL CITRAT 1. Tn khc, ch s 2. nh ngha INS 484 ADI = 0 - 50 mg/kg th trng, c thit lp ti k hp ln th 17 ca JECFA (1973). c tng hp bng cch ester ha acid citric vi cn stearylic thng mi, c th cha n-octadecanol v n 50% n-hexadecanol v t c cc yu cu di y. Ch phm thng mi c th c phn loi theo ch s x phng ha; hm lng v thnh phn ca cn stearylic; ch s iod; ch s acid v hm lng acid citric. Thnh phn gn ng

Cng thc cu to

3. M t 4. Chc nng 5. Tnh cht 5.1. nh tnh tan Cn stearylic Citrat 5.2. tinh khit Cc acid khc v ru khc Cht khng tan trong cloroform Ch 6. Phng php th 6.1. nh tnh Cn stearylic

Trong R1, R2 v R3 ln lt c th l C18H37 (stearyl), C16H33 (palmityl) hoc H Hp cht nhn, mu kem. Cht to phc kim loi, cht nh ha.

Khng tan trong nc v trong ethanol lnh, tan trong ethanol nng. Phi c phn ng c trng ca cn stearylic. Phi c phn ng c trng ca citrat. Khng c c cc acid khc ngoi acid citric v cc ru khc ngoi cc ru c mt trong ch phm cn stearylic thng mi. Khng c qu 0,5% (m t trong phn Phng php th). Khng c qu 2,0 mg/kg.

Thy phn 2 g mu th bng cch un hi lu trong 1 gi vi 50 ml dung dch natri hydroxyd (TS) thu c dung dch nc.

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Citrat

Lm mt v chit vi ether du ha. Ly lp ether du ha v cho bay hi trn a s hoc thy tinh. Phn khng bay hi phi c khong nng chy t 43o n 58o. Ly 5 ml dung dch nc thu c trong php th cn stearylic, thm 1 ml dung dch calci clorid (TS) v 3 git dung dch xanh da tri bromothymol (TS), acid ha nh bng dung dch acid hydrocloric long (TS). Thm dung dch natri hydroxyd (TS) n khi dung dch chuyn thnh mu xanh da tri sng, sau un si dung dch trong 3 pht, lc nh trong khi un. Trong dung dch xut hin kt ta trng, kt ta ny khng tan trong dung dch natri hydroxyd (TS) nhng tan trong dung dch acid acetic (TS). Ly 10 ml dung dch nc thu c trong php th cn stearylic, thm 1 ml dung dch thy ngn (II) sulfat (TS). un si hn hp v thm vi git dung dch kali permanganat (TS). Trong dung dch xut hin kt ta trng l mui thy ngn ca acid aceton dicarboxylic.

6.2. tinh khit Cht khng tan Ha tan khong 50g (cn chnh xc n mg) mu trong 400 ml cloroform. Lc dung dch qua phu lc thy tinh xp s 3 trong cloroform cn b (chnh xc n mg). Gi phu lc cn m, ra phn khng tan bng cloroform, sau sy kh ti 100o. 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.

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Ph lc 8 YU CU K THUT V PHNG PHP TH I VI TRIKALI ortho-PHOSPHAT 1. Tn khc, ch s Tripotassium phosphate; Tribasic potassium phosphate; Potassium phosphate. INS 340iii MTDI=70mg/kg th trng tnh theo Phospho t cc ngun thc phm Trikali phosphat; Trikali orthophosphat; Trikali monophosphat 7778-53-2 Dng khan: K3PO4 Dng hydrat: K3PO4.xH2O 212,27 (dng khan) Dng tinh th hoc ht khng mu hoc mu trng, khng mi; dng hydrat gm monohydrat v trihydrat m, cht nh ha, cht to phc kim loi

2. nh ngha Tn ha hc Ch s C.A.S. Cng thc ha hc Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan pH Kali Phosphat ortho-phosphat 5.2. tinh khit Gim khi lng khi nung

D tan trong nc, khng tan trong ethanol. 11,5 - 12,5 (dung dch 1/100). Phi c phn ng c trng ca kali. Phi c phn ng c trng ca phosphat. Phi c phn ng c trng ca ortho-phosphat. Dng khan: Khng c qu 3% (120oC trong 2 gi, sau 8000C trong 30 pht) Dng hydrat: Khng c qu 23% (120oC trong 2 gi, sau 800oC trong 30 pht) Khng c qu 0,2%. Khng c qu 10,0 mg/kg. Khng c qu 3,0 mg/kg (th theo hng dn ti JECFA monograph 1 - Vol.4 - phng php II).

Cc cht khng tan trong nc Fluorid Arsen

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Ch Khng c qu 4,0 mg/kg. 5.3. Hm lng Khng c thp hn 97,0% sau khi nung K3PO4 6. Phng php th 6.1. nh tnh Kali Thm 1 th tch dung dch natri hydrogen tartrat bo ha v 1 th tch dung dch ethanol vo dung dch mu th (1/100) v lc. Trong dung dch phi xut hin kt ta tinh th trng. Phosphat Thm 1ml dung dch acid nitric v 5ml dung dch nhm molybdat (TS) vo 5ml dung dch mu th (1/100) v un nng. Trong dung dch phi xut hin kt qu vng nht. ortho-phosphat Ha tan 0,1g mu th trong 10ml nc ct, acid ha nh dung dch bng acid acetic long (TS) v thm 1ml bc nitrat (TS). Trong dung dch phi xut hin kt ta mu vng 6.2. tinh khit Fluorid Cn 5 g mu th cho vo mt bnh cu ct 250 ml, thm 25 ml nc, 50 ml acid percloric, 5 git dung dch bc nitrat (1 trong 2) v vi vin bi thy tinh. Ni bnh cu vi 1 sinh hn c gn nhit k v mt ng mao qun sc kh, u nhit k v ng mao qun phi ngp trong dung dch cha trong bnh. Gn mt phu nh git cha nc hoc b sinh hi nc vo ng mao qun. t bnh trn tm aming c l sao cho 1/3 y bnh tip xc vi ngn la. Tin hnh ct n khi nhit trong bnh t 135o. Thm nc t phu vo bnh hoc cho hi nc qua bnh duy tr nhit trong bnh lun t 135o-140o. Tip tc ct n khi thu c 225-240 ml dch ct sau pha long dch ct thu c vi nc n 250 ml, lc u. Ly 50 ml dung dch ny cho vo mt ng Nessler 100 ml. Trong mt ng Nessler tng t khc ly 50 ml lm mu chng. Thm vo mi ng 0,1 ml dung dch natri alizarinsulfonat (1 trong 1000) lc v 1 ml dung dch hydroxylamin (1 trong 4000) mi pha, lc u. Thm tng git dung dch natri hydroxyd 0,05 N vo ng cha dch ct, va thm va khuy, n khi mu ng cha dch ct ging vi mu ng chng l mu hng nht. Sau thm vo mi ng ng chnh xc 1 ml dung dch acid hydrocloric 0,1 N v lc u. S dng buret chia vch 0,05 ml thm t t va dung dch thori nitrat (1 trong 4000) vo ng cha dch ct sau khi lc u mu ca dung dch chuyn v mu hng nht. Ghi th tch dung dch thori nitrat (1 trong 4000) s dng, thm chnh xc lng dung dch thori nitrat (1 trong 4000) nh vy vo ng

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Ch

6.3. nh lng

chng, lc u. Dng buret thm dung dch natri fluorid (TS) (10 g F / ml) vo ng chng cho mu hai ng ging nhau sau khi pha long v cng th tch. Lc u, yn cho bt kh thot ht trc khi so mu. Kim tra im tng ng bng cch thm 1-2 git dung dch natri fluorid (TS) vo ng chng. S thay i mu r rt s xy ra. Ghi li th tch dung dch natri fluorid (TS) s dng. Th tch dung dch natri fluorid (TS) s dng khng c qu 1 ml. - 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. Cn mu th (chnh xc n mg), khi lng tng ng vi 8g K3PO4 khan, ha tan vo trong 40 ml nc ct trong cc 400ml. Thm 100ml dung dch acid hydrocloric 1N. Sc khng kh kh carbon dioxyd vo dung dch, ch sao cho bt kh nh, thi gian sc 30 pht ui ht carbon dioxyd, y nh cc trnh mt dung dch trong khi sc. Ra np v thnh cc bng nc ct v cho in cc ca pH k thch hp vo dung dch. Chun dung dch bng dung dch natri hydroxyd 1N n im un ti pH~4, sau tnh th tch (A) dung dch acid hydrocloric 1N s dng. Bo v dung dch trnh hp th carbon dioxyd t khng kh v tip tc chun vi dung dch natri hydroxyd 1N n im un ti pH~8,8. Tnh th tch (B) dung dch natri hydroxyd 1N tiu tn trong chun ny. Nu (A) 2(B), mi ml natri hydroxyd 1N trong th tch (B) tng ng vi 212,3mg K3PO4. Nu (A) < 2(B), mi ml natri hydroxyd 1N trong hiu th tch (A)-(B) tng ng vi 212,3mg K3PO4.

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Ph lc 9 YU CU K THUT V PHNG PHP TH I VI CC MUI AMONI CA ACID PHOSPHATIDIC 1. Tn khc, ch s Ammonium phosphatides, Emulsifier YN, Mixed ammonium salts of phosphorylated glycerides; INS 442 ADI = 0 -30 mg/kg th trng Ch phm ny ch yu gm hn hp cc hp cht amoni ca cc acid phosphatidic bt ngun t cht bo n c (thng l du ht ci hydrogen ha mt phn). L mt mono- hoc diglycerid c th c gn phosphor. Tuy nhin, hai ester phosphor c th lin kt vi nhau dng phosphatidyl phosphatid. Ch phm c sn xut bng cch phn hy cht bo bng glyceryl, sau phosphoryl ha bng phosphor pentoxyd, v trung tnh ha bng amoniac. L mt hng thng mi c th c quy nh thm nhng chi tit c th nh hm lng nc, tp cht khng tan trong hexan, cht v c khng tan trong hexan, gi tr pH v hm lng triglycerid.

2. nh ngha

Cng thc cu to (thnh phn gn ng) Trong R c th l mt gc mono- hoc di-glycerid 3. Cm quan Cht na rn trn nhn. 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan Khng tan trong nc, tan mt phn trong ethanol v aceton; tan trong cht bo. Phosphat Phi c phn ng c trng ca phosphat. Acid bo Phi c phn ng c trng ca acid bo. Glyceryl Phi c phn ng c trng ca glyceryl. 5.2. tinh khit Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng Hm lng phosphor khng c nh hn 3,0% v khng ln hn 3,4% theo khi lng; hm lng nitrogen amoni khng c nh hn 1,2% v khng ln hn 1,5%.

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6. Phng php th 6.1. nh tnh Phosphat Nung 1 g mu th vi 2 g natri carbonat khan. ngui v ha tan cn trong 5 ml nc v 5 ml acid nitric. Thm 5 ml amoni molybdat (TS) v un ti si. Xut hin ta mu vng. Acid bo un hi lu 1 g ch phm th trong 1 gi vi 25 ml kali hydroxyd 0,5 N trong ethanol. Amoniac bay ra t pha u ca ng sinh hn hi lu, nhn ra bng mi ca n v phn ng trn giy quy m. Lm lnh cn ti 0oC, ta x phng kali to thnh. 6.2. tinh khit 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 Phosphor Phng php quang ph hp th Thuc th v dung dch lm vic Acid sulfuric: t trng tng i 1,84 Acid nitric: t trng tng i 1,42 Acid perchloric: 60%, t trng tng i 1,54 Dung dch vanadat-molybdat: ha tan ring trong nc 20 g amoni molybdat v 1 g amoni vanadat. Trn hai dung dch, thm 140 ml acid nitric c v pha long ti 1000 ml bng nc. Lc u. Dung dch phosphat chun: Dung dch gc: ha tan 3,8346 g kali dihydro phosphat, lm kh 110C, vo trong nc v pha long ti 1000 ml; 1 ml dung dch ny = 2,0 mg P2O5. Dung dch chun lm vic phosphat: Chuyn 50,0 ml dung dch phosphat chun (1ml = 2,0 mg P2O5) vo bnh nh mc 500 ml v pha long ti vch bng nc v lc u. 1 ml dung dch chun lm vic phosphat = 0,2 mg P2O5. Tin hnh Cn chnh xc 1,5 ti 1,6 g mu th vo mt bt thy tinh nh v cho vo trong bnh Kjeldahl 300-ml cha 5 ml acid sulfuric v 10 ml acid nitric. un nng bnh, u tin un nh v lc lin tc, sau un mnh hn trn ngn la trn. Thm nhiu ln ti cc thi im khc nhau mi ln mt lng acid nitric, lm ngui

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bnh trc khi thm, v tip tc un ti khi thu c dch phn hy trong v c mu vng kim. ngui, thm 5 ml acid percloric 60% v tip tc oxy ha ti khi to thnh khi acid trng trong bnh. Li lm ngui v thm 5 ml nc v tip tc un ti khi ht khi trng bay ln. ngui, pha long cn thn bng nc, li ngui v chuyn ton lng sang bnh nh mc 500-ml. Pha long ti vch bng nc v lc u (Dung dch th). Lm mt mu trng phn hy ng theo cch nh trn nhng khng cho mu th v s dng cng th tch acid nh dng v c ha t mu th (Dung dch trng phn hy). Cho vo cc bnh nh mc 100-ml ring, thm t buret: (a) 25,0 ml dung dch chun lm vic phosphat (= 5,0 mg P2O5), (b) 30,0 ml dung dch chun lm vic phosphat (= 6,0 mg P2O5), (c) 25 ml dung dch th cha t 5 n 6 mg P2O5 Cho vo mi bnh cha phosphor chun, (a) v (b), mt lng dung dch trng phn hy c cng th tch nh dung dch cho vo (c), b i vi vt phosphor c th c do thuc th phn hy acid m n c th c trong dung dch th. Cho vo mi bnh 25 ml thuc th vanadat-molybdat, lc u, pha long bng nc ti gn 100 ml, lc u, iu chnh nhit ca dung dch ti 20C, pha long ti vch bng nc v lc u. Sau 10 pht, o hp th ca c dung dch 6 mg P2O5 v dung dch th, mu trng l dung dch cha 5 mg cht chun. Dng cc o 1 cm v o bc sng 420 nm, hoc knh lc Ilford 604 nu dng my so mu quang in. Tnh ton
A t 0,873 % phosphor = 5 + A m 6mg

Trong : At = hiu hp th gia chun 5 mg v dung dch th A6 mg = hiu hp th gia chun 6 mg v 5 mg m = khi lng mu th (g) Xc nh Thit b ct ko hi nc nitrogen amoni Thit b gm mt bnh 2-L y nt cao su, lp xuyn qua nt c mt ng thy tinh khong 7,5 cm, c lp sao cho u di ca ng gn st y bnh, v mt on ng hnh ch L ngn hn lp

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sao cho ng cha ngang khong 6 mm bn di b mt ca nt, hot ng nh ng ra ca lung hi nc. Bnh c thm nc ct acid ha nh bng acid sulfuric long (TS) ti khong 2/3 th tch bnh v vi mnh thy tinh xp trnh tro khi hn hp trong bnh si mnh. Bnh ny dng lm ngun cung cp hi nc. C th lp mt vi vo bnh nu mun d dng b sung nc vo bnh gia cc ln xc nh. ng ra ca hi c ni qua mt by ngng ti u vo ca u ct hi, ni vo mt bnh cu y trn c ngn 1-L c bnh c B34. u ct phi sao cho ng u vo ca hi gn chm y bnh 1-L v u ra phi ni vi hai by trnh bn, mt gn nh ca bnh 1-L v ci kia gn nh ca mt sinh hn thng ng, ni c B19, ni vi u ct. Sinh hn thng phi lp vi ng u ra ko di, chm ti y ca mt bnh nn 500-ml. Thuc th Dung dch acid boric 2% (kl/tt) trong nc Dung dch natri hydroxyd 40% (kl/tt) trong nc Acid hydrocloric 0,02 N Hn hp ch th: Trn 5,0 ml dung dch xanh lc bromocresol 0,1% (kl/tt) trong alcol v 2,0 ml dung dch methyl 0,1% (kl/tt) trong alcol v pha long ti 30 ml bng alcol 95%. Dch silicon 200/50 MS Tin hnh Lp v cho hi nc i qua thit b. Cn chnh xc khong 0,2 g mu th mui amoni ca acid phosphatidic trung tnh vo trong mt l thy tinh nh (ng knh khong 18 mm, su khong 12 mm). Chuyn l v mu th cn vo bnh ct v thm khong 250 ml nc ct. Ni u ct v by chng bn vi bnh ct v sinh hn ng, lp sinh hn sao cho u ra ngp di b mt ca 10 ml acid boric 2% v 1 ml hn hp ch th ng trong bnh nn 500-ml. Thm vo bnh ct qua phu gn bng mt ng cao su ngn vo ng dn hi nc vo, 75 ml natri hydroxyd 40%, v ra vo trong bng nc ct. B phu v ni ng vo ca hi vi ngun cung cp hi. (C th cho natri hydroxyd vo bnh qua phu c kha, lp vi bnh ct nu mun v ra pha trong bng nc ct. Nu lm nh vy phi duy tr mt lp cht lng trn phu trong sut qu trnh thm v ct). Ct ko mnh bng hi nc hn hp trong bnh ct v thu ly 200 ml dch ct vo trong acid boric. Trong qu trnh ct khuy nh bnh ct nu cn, trnh

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mu th b lng ng xung quanh b mt pha trn ca bnh. Khi thu lng dch ct theo yu cu, h thp bnh hng, ngng cung cp hi nc v ra pha trong ca sinh hn v bn ngoi u pha di vi mt lng nh nc ct, gp dch ra vo bnh hng. Chun hn hp trong bnh hng bng acid hydrocloric 0,02 N. Lm t nht mt mu trng tin hnh theo cng cch nhng khng c mu th. Trong qu trnh ct c th c bt xut hin ca hn hp trong bnh ct. Nu c, thm 2 git dch silicon vo bnh ct ti thi im cho mu th; v mt lng tng t c cho vo mu trng. Tnh ton Mi ml HCl 0,02 N = 0,2802 mg nitrogen. % Nitrogen =

Vt V o 28,02 mt

Trong Vt = th tch dung dch HCl 0,02 N dng chun mu th (ml) Vo = th tch dung dch HCl 0,02 N dng chun mu trng (ml) mt = khi lng mu th (mg).

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Ph lc 10 YU CU K THUT V PHNG PHP TH I VI SUCROSE ACETAT ISOBUTYRAT 1. Tn khc, ch s 2. nh ngha SAIB INS 444 Hn hp c to thnh bng cch este ha sucrose thc phm vi anhydrid acetic v anhydrid isobutyric v chng ct. Hn hp bao gm tt c cc este vi t l phn t ca acetat v isobutyrat l khong 2:6 Sucrose diacetat hexaisobutyrat 137204-24-1; 27216-37-1; 126-13-6 C40H62O19 i vi sucrose diacetat hexaisobutyrat

Tn ha hc Ch s C.A.S Cng thc ha hc Cng thc cu to

Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Ch s khc x Khi lng ring Hp th hng ngoi

Trong R1 = -CH2OCOCH3 R2 = -CH2OCOCH(CH3)2, hoc -OCOCH(CH3)2 832 - 856 C40H62O19 = 846,9 Dng lng mu vng nht, khng cn v c mi nh Cht nh ha, cht iu chnh m c, cht to c i vi ung khng cn

Khng tan trong nc, tan trong hu ht dung mi hu c. n (40, D): 1,4492 - 1,4504 d (25, 25): 1,141 - 1,151 Ph hng ngoi ca mu th phn tn trong kali bromid tng ng vi ph hng ngoi chun trong Ph lc.

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5.2. tinh khit Ch s acid Khng c qu 0,2 (s dng 50 g mu th). Ch s x Gia 524 v 540 (s dng 1 g mu th) phng ha Triacetin Ch 5.3. Hm lng C40H62O19 Khng c qu 0,1%. Khng c qu 2 mg/kg. Khng c nh hn 98,8% v khng c qu 101,9%.

6. Phng php th 6.1. tinh khit Triacetin S dng phng php sc k kh nh sau: Dng c, thit b: My sc k kh c trang b detector ion ha ngn la Ct: thp khng r, di 1,5 m, ng knh trong l 3,2 mm. Chun b mu: Pha long mu th bng cch cho thm mt th tch carbon disulfid tng ng. iu kin: Pha tnh: SE-30, 3% Pha rn: Chromosorb AW-DMCS, 80-100 mesh Kh mang: Heli Tc kh: 20 ml/min Nhit - Ct: Chng trnh nhit : ngay sau khi bm mu, nhit ct tng t 1000C ln n 3000C, tc tng 100C/pht. - Nhit bung bm: 3000C Th tch bm mu: 1 l - 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.

Ch

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6.2. nh lng S dng ch s x phng ha, tnh % C40H62O19 theo cng thc:

Ph hng ngoi

Trong SV = ch s x phng ha Sucrose acetat isobutyrat

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Ph lc 11 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI NHA CY 1. Tn khc, ch s 2. nh ngha Ester gum; INS 445 Ester phc hp ca triglycerol v diglycerol vi cc acid t nha g thng thu c nh qu trnh trch ly gc cy thng lu nm bng dung mi sau c tinh ch trong h dung mi lnglng. Loi tr khi cc yu cu k thut ny l cc dn xut ca gum nha, dch tit ra t cy thng sng, nha du thng cao v sn phm ph ca qu trnh sn xut bt giy. Sn phm cui cng gm c 90% acid nha v 10% cht trung tnh (cht khng acid). Phn acid nha l mt phc hp ca cc ng phn acid monocarboxylic diterpenoid c cng thc thc nghim in hnh l C20H30O2, trong thnh phn chnh l acid abietic. Glycerol ester ca nha g thng c tinh ch bng phng php bc hi nc hoc chng ct ngc dng. 8050-30-4 Cht rn mu vng n mu h phch nht Thnh phn base trong ko cao su, cht nh ha, cht n nh, cht n, cht iu chnh hng liu trong ung

M s C.A.S. 3. Cm quan 4. Chc nng

5. Yu cu k thut 5.1. nh tnh tan Khng tan trong nc, tan trong aceton Hp th hng Ph hng ngoi ca lp mng mu c t trn tm kali bromid ngoi tng ng vi ph hng ngoi in hnh di y Alcol nha v Phi c phn ng c trng ca alcol nha v glycerol nha. glycerol nha 5.2. tinh khit Khng c mt t yu cu (m t trong phn phng php th). nha du cao (Lu hunh) T trng d (20, 25): Khng c nh hn 0,935 khi xc nh trong dung dch 50% trong d-limonen (97%, im si 175,5 - 176,00C, d (20, 4): 0,84) im nng 82 - 900C (m t trong phn Phng php th) chy Ch s acid 3-9

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Ch s hydroxyl 15 - 45 (m t trong phn Phng php th ) Ch Khng c qu 2,0 mg/kg. 6. Phng php th 6.1. nh tnh Alcol nha v Cc nhm ester phc tp trong ester ca glycerol nha g thng glycerol nha c kh bng phn ng vi mt hydrid kim loi (natri bis (2-methoxy-ethoxy) nhm dihydrid) trong dung dch toluen to thnh mt hn hp ca alcol nha v glycerol nha. Cht kh d sau c thy phn bng dung dch acid to thnh hai pha. Sc k kh ca pha toluen to ra mt sc k ca alcol nha cu thnh ester nha g thng v c th phn bit vi cc ester gum v ester nha du cao. Nha g c th phn bit c vi nha du cao da vo t l ca abietyl v dehydroabietyl alcol: trong nha g abietyl chim u th, trong khi nha du cao, dehydroabietyl chim u th. Nha g c th phn bit c vi gum rosin da vo t l ca isopimaryl v palustryl alcol: trong nha g isopimaryl chim u th, trong khi gum rosin, palustryl chim u th. Sc k ca pha trung tnh trong nc trn ct khc kim tra s xut hin ca glycerol. Thit b, dng c: - Sc k kh c trang b detector ion ha ngn la - Ct sc k I: Column I: DB-1 methyl silicon (ngoi quan v lin kt cho), ct mao qun di 15 m, ng knh trong 0,53 mm, dy 1,5 m, di nhit 60 - 300/3200C (V d: J & W Scientific Inc,. Cat. No. 125-1012). C ng dn bm hi nhanh, trc tip. - Ct sc k II:: DB-Wax polyethyleneglycol (ngoi quan v lin kt cho), ct mao qun di 15 m, ng knh trong 0,53 mm, dy 1,0 m, di nhit 20 - 2300C (V d: J & W Scientific Inc,. Cat. No. 125-7012). - B ghi: 0 - 1V - Syringe: 1 l - Pipet: 3,0 ml; 5,0 ml - Bnh Erlenmeyer: 25 ml - L: 17 ml - Cn phn tch: cn chnh xc n 0,001 g - ng ly tm c vch nh mc: 15 ml - My ly tm: c th t tc 3.200 vng/pht - Thanh khuy: c bao bng Teflon, kch thc 1 inch

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- My khuy t Ha cht, thuc th: - Toluen tinh khit - Natri Vitride TM [natri bis (2-methoxyethoxy) nhm dihydrid, thng xp x 70% trong toluen (khong 3,5 mol/l)] (Tp on ha cht Fluka ., Hauppage, NY, M). Ly 10 ml cho vo bnh nh mc dung tch 100 ml. Pha n th tch 100 ml bng toluen v lc u. - Dung dch thy phn: Thm t t 50 ml dung dch H2SO4 m c, tinh khit vo 200 ml nc ct ng thi khuy trong chu nc . Lm ngui ti nhit phng. - Dung dch phenolphthalein: 1% trong ethanol. - Dung dch NaOH: ha tan 16 g NaOH trong 70 - 80 ml nc ct v lm ngui ti nhit phng. Pha ti 100 ml bng nc ct v lc u. trong bnh polyethylen. - 1, 4- Butandiol: 99% - Glycerol: 99% - Dung dch ni chun: Cn 0,1 g 1, 4- butandiol cho vo bnh nh mc dung tch 100 ml. Pha ti th tch 100 ml bng nc ct v lc u. - Dung dch glycerol: Cn 0,1 g 1, 4- butandiol v 0,1 g glycerol cho vo bnh nh mc dung tch 100 ml. Pha ti th tch 100 ml bng nc ct v lc u. iu kin chy sc k kh: I (alcol nha) Nhit : - Ct: 1900C, ng nhit - u vo: 2500C - Detector: 2500C Tc dng: - Kh mang (He): 30 ml/pht ti 63 psi - H2: 30 ml/pht - Khng kh: 240 ml/pht II (glycerol) Nhit : - Ct: t theo chng trnh, 120 - 2000C vi tc 60C/pht - u vo: 2500C - Detector: 2500C Tc dng:

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- Kh mang (He): 30 ml/pht ti 63 psi - H2: 30 ml/pht - Khng kh: 240 ml/pht Cch tin hnh I (i vi nha): Cn 250 - 300 mg mu cho vo bnh Erlenmeyer dung tch 25 ml c thanh khuy t. Dng pipet ly 5,0 ml toluen cho vo bnh v khuy t cho n khi mu c ha tan. Ly 5,0 ml Natri VitrideTM cho vo bnh, y np bnh v khuy trong 30 pht. M np ra ng thi khuy, dng pipet ly 3,0 ml dung dch thy phn cho vo bnh. Tip tc khuy trong 3 pht. Chuyn hn hp trong bnh vo ng li tm, y np v lc u. Cho thot kh v ly tm vi tc 2.800 - 3.200 vng/pht trong 5 pht. Bm 0,5 l lp trn vo thit b sc k kh vi cc iu kin k thut t v ghi sc k . So snh vi cc sc k c biu th pha di xc nhn th t thi gian lu ca cc alcol nha. Cch tin hnh II (i vi glycerol) Dng pipet hoc syringe hypodermic, loi b lp toluen v mt phn dung dch nc v li khong 2 ml dung dch nc trong ng ly tm. Thm 1 git dung dch phenolphthalein v trung ha bng dung dch NaOH. Cc mui nhm s kt ta. Dng pipet ly 5 ml dung dch ni chun cho vo ng, pha n 15 ml bng nc ct, y np, lc v ly tm vi tc 2.800 - 3.200 vng/pht trong 5 pht. Bm 1 l dch trong trn b mt vo thit b sc k kh vi cc iu kin k thut t v ghi sc k . Bm 1 l dung dch glycerol v ghi sc k . o cc thi gian lu ca bt k cc pic so vi 1,4- butandiol. So snh cc thi gian lu vi thi gian lu ca glycerol. 6.2. tinh khit Khng c Khi cc hp cht hu c c cha lu hunh c gia nhit vi s nha du cao c mt ca natri format, lu hunh chuyn ha thnh hydrogen (Lu hunh) sulfid c pht hin bng cch s dng giy ch acetat. Php th dng tnh cho thy vic s dng nha du cao thay cho nha g. Thit b, dng c: - ng th: loi tiu chun, kch thc 10 x 70 mm, lm bng thy tinh chu nhit. - n Bunsen: Thng dng c nh dng ngn la nh Ha cht, thuc th: - Dung dch natri format: ha tan 20 g natri format tinh khit (NaOOCH) trong 100 ml nc ct. - Giy th ch acetat: dng thng mi c sn ca hu ht cc nh cung cp ha cht.

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Cch tin hnh: Cn 40 - 50 mg mu cho vo ng th v thm 1 -2 git dung dch natri format 20% (khi lng/th tch). di giy th ch acetat trn ming ng th. Gia nhit ng th bng ngn la cho n khi khi c hnh thnh v tip xc vi giy th. Tip tc gia nhit trong 2 - 5 pht. Khng c c m en ca ch sulfid hnh thnh, nu c m en th chng t s c mt ca cc hp cht cha lu hunh. (Gii hn pht hin: 50 mg/kg lu hunh). im nng chy im nng chy c nh ngha l nhit ti mt a mu c gi bn trong mt chic nhn nm ngang gim xung mt khong 25,4 mm di trng lng ca mt cu thp khi mu c gia nhit vi tc quy nh trong chu cch thy hoc cch glycerol. Thit b: Thit b c minh ha trong Hnh 1 v Hnh 2 gm c cc b phn c m t trong cc phn di y. Chic nhn: c lm bng ng thau ph hp vi kch thc trnh by trong Hnh 1a. Nu cn, vng c th c gn bng cch hn hoc cch khc thun tin vi mt dy ng thau loi 13 B&S (ng knh 0,06 - 0,08 inch hoc 1,52 - 2,03 mm) c trnh by Hnh 2a. Qu cu: thng dng qu cu c lm bng thp c ng knh 3/8 inch (9,53 mm), khi lng 3,45 - 3,55 g. Hng dn chnh tm qu cu: Hng dn chnh tm qu cu c lm ng thau v c hnh dng, kch thc chung c minh ha trong Hnh 1c, c th c s dng nu mun. Dng c cha: S dng bnh thy tinh chu nhit nh cc Griffin dng ln dung tch 800 ml, ng knh khng c nh hn 3,34 inch (8,5 cm) v chiu su tnh t y n phn loe ra khng c nh hn 5 inch (12,7 cm). Dng c gi chic nhn v nhit k: Bt k dng c hu hiu no ph hp vi cc yu cu sau dng gi vng v nhit k u c th c s dng: (1) Vng nhn c th c gi v tr nm ngang; (2) khi s dng thit b m t trong Hnh 1d, y ca vng trn nm trn v cch tm nm ngang pha di 1 inch (25,4 mm), b mt y ca tm nm ngang cch t nht 0,5 inch (13 mm) v khng c qu 0,75 inch (18 mm) so vi y ca dng c cha bn trn, chiu su

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ca dch lng trong dng c cha khng c nh hn 4,0 inch (10,2 cm); (3) khi s dng thit b a ra Hnh 1e, y ca vng trn cch 1 inch (25,4 mm) so vi y ca dng c cha pha trn, vi im di cng ca thanh ngh t trn y ca dng c cha v chiu su ca dch lng trong dng c cha khng c nh hn 4,0 inch (10,2 cm) nh m t Hnh 1 a, b v c; (4) trong c hai dng c, nhit k c treo ln sao cho khong cch t y ca bu nhit k n y ca chic nhn khong 0,5 inch (13 mm) nhng khng c chm vo nhn. Nhit k thy ngn: Ph thuc vo im nng chy ca mu, s dng hoc nhit k ASTM 15C (-2 n 800C) i vi loi c im nng chy thp hoc nhit k ASTM 16C (30 - 2000C) i vi loi c im nng chy cao. My khuy: S dng my khuy thch hp vi tc 500 - 700 vng/pht. m bo phn tn nhit u trong mi trng nng, hng ca trc quay nn y dch lng ln pha trn. (Xem Hnh 2d ). Chun b mu: Ly mu i din gm cc mnh va mi b v c b mt cha b oxy ha. No lp trn b mt mu thu cc mnh v ngay trc khi s dng, trnh bao gm cc bt mn hoc bi. Lng mu ly thng t nht gp i lng cn cho s chic nhn yu cu nhng khng c nh hn 40 g. Ngay sau lm nng chy mu trong dng c cha sch, s dng t sy, tm gia nhit hoc cch ct hoc cch du ngn qu nhit cc b. Trnh hnh thnh bt kh trong mu nng chy, khng c gia nhit trn nhit cn thit mu d dng m khng bao gm bt kh. Thi gian tnh t lc bt u gia nhit cho n lc mu khng qu 15 pht. Ngay trc khi y cc chic nhn, gia nhit mu trc n nhit ti mu c rt. Trong khi rt, cc vng trn c gi trn tm ng thau. Rt mu vo cc vng trn tha cho ngui. Lm ngui t nht 30 pht v sau ct b phn mu tha bng dao h nng hoc dao bay. S dng dng c cha sch v mt mu mi nu php th c lp li. Cch tin hnh: Mu c im nng chy trn 800C: Rt glycerol vo bnh thy tinh ti su khng c nh hn 4 inch (10,2 cm) v khng c qu 4,25 inch (10,8 cm). Nhit ban u ca chu l 320C. i vi nha thng (gm c nha g), glycerol cn c

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Ch s hydroxyl

lm ngui n nhit khng c nh hn 450C di im nng chy d on trc nhng khng c thp hn 350C. V tr trc ca trc khuy gn thnh sau ca dng c cha, vi cc li dao khng chm vo thnh v phn di ca cnh dao cch nh ca vng 0,75 in (18 mm). Tr khi c hng dn chnh tm cu c s dng, lm lm nh trung tm ca mu bng cch nhn qu cu hoc mt que trn c h nng dng cho mu cng, vo mu ti thi im ny. Treo vng trn c cha cc mu trong chu glycerol b mt di ca vng trn y cch b mt ca tm ngang thp hn l 1,0 inch (25,4 mm) (xem Hnh 1d), t nht 0,5 inch (13 mm) v khng qu 0,75 inch (18 mm) trn so vi y ca bnh thy tinh, hoc 1,0 inch (25,4 mm) trn so vi y ca dng c cha (xem Hnh 2e). t qu cu trong glycerol nhng khng c t ln trn mu th. Treo nhit k ASTM 16C trong glycerol sao cho y bu ca nhit k cch y chic nhn khong 0,5 inch (13 mm) nhng khng c chm vo chic nhn. Duy tr nhit ban u ca glycerol trong 15 pht v sau dng kp thch hp t qu cu vo trung tm b mt trn ca mu trong chic nhn. Bt u khuy v tip tc khuy vi tc 500 - 700 vng/pht cho n khi hon tt vic xc nh. Gia nhit theo tc tng nhit ca glycerol 50C/pht, trnh nh hng ca cn bng cch s dng cc tm chn nu cn thit. [Ch : Tc tng nhit s khng i v s khng c tnh theo gi tr trung bnh trong ton b qu trnh th. Loi b tt c cc php th trong tc tng nhit vt qu 0.5C cho bt k qu trnh no sau 3 pht u]. Ghi li im nng chy, chnh l nhit trn nhit k ti mu chm vo tm nm ngang pha di (xem Hnh 1d) hoc y ca dng c cha (xem Hnh 2e). Khng hiu chnh thn nhit k. Mu c im nng chy di 800C: thc hin theo cch tin hnh trn, ngoi tr s dng nhit k ASTM 15C v s dng nc va un si c lm mt ti 50C to mi trng nhit. i vi nha thng (gm c nha g), s dng nc c lm ngui n nhit khng c nh hn 450C di im nng chy d on trc nhng khng c thp hn 50C. Ch s hydroxyl c nh ngha l s mg KOH dng trung Ha carbamat sulfonyl c kh nng kt hp vi cc nhm hydroxyl trong 1 g mu. Cc nhm hydroxyl trong hp cht hu c phn ng nhanh vi p - toluensulfonyl isocyanat d (p - TSI) trong dung mi tr to thnh carbamat sulfonyl. Sau phn ng

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ca thuc th d vi nc to thnh p - toluensulfonamid, carbamat sulfonyl c tnh acid c chun bng trc quan hoc thit b chun bng dung dch KOH trong methanol, pht trin im tng ng bng o th hoc bng ch th mu. Cc hiu chnh c thc hin i vi mu trng v s c mt ca bt k cc hp cht c tnh acid. Thit b, dng c: - Thit b chun t ng: s dng thang o pH 0 - 14, t tc 4 (khong 2 ml/pht) v mt buret 20 ml. - in cc thy tinh - in cc calomel - Bnh Erlenmeyer: dung tch 250 ml c khp ni thy tinh hnh trn - Sinh hn Allihn: di 300 mm c khp ni thy tinh hnh trn khp vi bnh Erlenmeyer. Ha cht, thuc th: - Acid benzoic, t tiu chun cht chun gc - Methanol, tinh khit, dng khan - Toluen, tinh khit, sy kh qua ry phn t 3 - KOH 0,1 N pha trong methanol: ha tan 7 g KOH trong 1 lt methanol. Chun ha bng cch cn chnh xc n 0,0001 g, khong 0,12 g acid benzoic vo cc dung tch 150 ml. Cho thanh khuy t v 80 ml hn hp toluen/methanol theo t l 1/1. Khuy u dung dch trong khi chun vi dung dch KOH trong methanol. Dung dch KOH cn phi cho xung di b mt ca dung dch, cch xa in cc. Tnh ton nng dung dich KOH trong methanol da theo phng trnh tnh ton c biu th di y. - Tetrahydrofuran (THF): sy qua m v cho qua ry phn t 3 - p-Toluenesulfonyl isocyanat (p-TSI) - Dung dch p-Toluenesulfonyl isocyanat: khong 0,22 M hoc 4,5 mEq/20 ml dung dch. Dng pipet ly 15 ml p-Toluenesulfonyl isocyanat (p-TSI) cho vo 500 ml tetrahydrofuran (THF) kh v lc u. Ch : V isocyanat c bn cht rt c nn khi chun b v s dng dung dch isocyanat phi c tin hnh trong t ht. Cn phi eo gng tay khi cm vo isocyanat v dung dch ca isocyanat trnh tip xc vi da.

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- Dung dch ch th bromocresol ta: cho 0,1 g bromocresol ta vo 18,5 ml dung dch NaOH 0,1 N v pha ti th tch 250 ml bng nc ct. Cch tin hnh: Cn 1 - 1,5 g mu chnh xc n 0,0001 g cho vo bnh Erlenmeyer kh dung tch 250 ml c ni vi khp ni thy tinh hnh trn. Dng pipet ly 10 ml dung dch THF kh cho vo bnh v ha tan mu. Dng pipet ly 20 ml dung dch p - TSI cho vo bnh v lc u. Cho vi ht bt, ni vi sinh hn v bnh trn tm gia nhit. Gia nhit n khi si vi sinh hn ngc trong 10 pht. Trong qu trnh chng ct b sung thm nc thch hp qua sinh hn: Mu trng Mu th Chun vi ch th mu 2 2 Chun bng o th 1 1 Ly bnh v sinh hn ra khi tm gia nhit v ngui ti nhit phng. Ra sinh hn bng 5 ml THF. Tho sinh hn ra khi bnh v chuyn ton b phn dch trong bnh vo cc dung tch 150 ml vi s tr gip ca 50 ml THF. Cho thanh khuy t v nhng cc in cc vo hoc thm 20 git dung dch ch th. Va khuy va chun bng thit b chun o th hoc chun vi ch th mu trc quan vi dung dch KOH trong methanol. Dung dch KOH cn phi cho xung di b mt ca dung dch cch xa in cc. Xc nh th tch dung dch KOH dng chun mu th v chun mu trng c thc hin qua tt c cc bc tin hnh trn. Tnh kt qu: Nng ca dung dch KOH trong methanol (N):

Trong : W: Khi lng acid benzoic

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122,1: ng lng ca acid benzoic KOH: S ml dung dch KOH trong methanol dng chun Ch s hydroxyl trong mu:

Asen

Ch

Trong : A: S ml dung dch KOH dng chun mu th B: S ml dung dch KOH dng chun mu trng N: Nng dung dch KOH trong alcol 56,1: S mg KOH/mEq AV: Ch s acid Chun b dung dch mu nh sau: Cn chnh xc 1 g mu v cho vo bnh Kjeldahl, t u mt ng ca bnh trong qu bu bc hi c gn vi my ht nc, thm 5 ml dung dch H2SO4 v 4 ml dung dch hydrogen peroxyd 30% v ph mu trn ngn la nh. Tip tc thm 2 ml peroxyd trong 2 ln, phn ng lng cn gia hai ln thm cho n khi tt c cht hu c b ph hu, khi ca acid H2SO4 bc ra mnh v dung dch tr ln khng mu. Duy tr cc iu kin xy ha ti tt c cc thi im trong qu trnh ph mu bng cch b sung peroxyd mi khi hn hp chuyn mu nu hoc ti mu. (Lng peroxyd cn ph hon ton mu s thay i nhng trong mt s trng hp mc ti a c th l 200 ml, iu ny ph thuc vo trng thi t nhin ca mu). Lm ngui, ch thm 10 ml nc, li cho bc hi n khi to thnh khi mnh v lm ngui. Chuyn dung dch ti bnh to arsin, ra bnh Kjeldahl v qu bu bng nc, thm nc ra vo bnh v pha ti 35 ml bng nc. Dung dch ny phi p ng c cc yu cu ca Gii hn th arsenic. Cn chnh xc 5 g mu v cho vo a s hoc bt s, un trn tm gia nhit cho n khi than ha hon ton, sau gia nhit trong l nung 4800C 8 gi hoc qua m, ngui. Thm cn thn 5 ml acid nitric, lm bay hi n kh trn tm gia nhit, sau nung trong l nung 4800C trong 15 pht, v lm ngui. Trch ly tro trong 10 ml nc trong 2 ln, lc mi phn dch chit vo mt bnh gn. Lc qua bt k cht khng tan trn dng c lc bng 6 ml dung dch amoni citrat TS, 2 ml hydroxylamin hydrochlorid TS v 5 ml nc, thm nc ra c lc vo bnh gn. Tip tc theo hng dn trong phn Cch tin hnh trong th Gii hn, bt u vi Thm 2 git phenol TS .... vo bnh gn, s dng 10 g ion ch (Pb) trong mu i chng.

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Ph hng ngoi

Glycerol ca nha g thng

Sc k kh ca cc alcol nha thng

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87

Thit b, dng c - im nng chy

Hnh 1

Hnh 2

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Ph lc 12 YU CU K THUT V PHNG PHP TH I VI DINATRI DIPHOSPHAT 1. Tn khc, ch s Disodium pyrophosphate; Disodium dihydrogen diphosphate; acid sodium pyrophosphate. INS 450i MTDI=70mg/kg th trng tnh cho Phospho t cc ngun thc phm Disodium dihydrogen diphosphate; Disodium dihydrogen pyrophosphate 7758-16-9 Dng khan: Na2H2P2O7

2. nh ngha Tn ha hc

Ch s C.A.S. Cng thc ha hc Khi lng phn 221,94 t 3. Cm quan Dng tinh th hoc ht mu trng. 4. Chc nng Cht nh ha, m, cht to phc kim loi, cht to xp 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc. pH 3,7-5,0 (dung dch 1/100). Natri Phi c phn ng c trng ca natri. Phosphat Phi c phn ng c trng ca phosphat. 5.2. tinh khit Gim khi lng Khng c qu 0,5% (105oC trong 4 gi) khi sy kh Cc cht khng Khng c qu 1%. tan trong nc Fluorid Khng c qu 10,0 mg/kg Arsen Khng c qu 3,0 mg/kg Ch Khng c qu 4,0 mg/kg 5.3. Hm lng Khng thp hn 95,0%. 6. Phng php th 6.1. tinh khit

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Cc cht khng tan trong nc 6.2. nh lng

Ha tan 10 g mu th trong 100 ml nc nng. Lc qua mt chn lc cn b. Ra cn khng tan trn chn lc bng nc nng. Sy kh ti (105oC trong 2 gi). ngui v cn. Cn 400mg (chnh xc n mg) mu th c sy kh trc ti (105oC trong 4 gi), ha tan trong 100 ml nc trong cc 400 ml. Chnh pH dung dch v 3,8 bng dung dch acid hydrocloric long (TS) hoc dung dch natri hydroxyd (TS), dng pH k xc nh pH dung dch. Sau thm 50 ml dung dch km sulfat (125 g ZnSO4.7H2O ha tan trong nc, pha long n 1000ml, lc v chnh pH v 3,8), yn trong khong 2 pht. Chun acid c gii phng bng dung dch natri hydroxyd 0,1 N n khi pH dung dch li tr v 3,8. Sau mi ln thm natri hydroxyd gn n im tng ng, cn mt thi gian nu c kt ta km hydroxyd th kt ta c th tan ht. Mi ml natri hydroxyd 0,1 N tng ng vi 11,10mg Na2H2P2O7.

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Ph lc 13 YU CU K THUT V PHNG PHP TH I VI CALCI POLYPHOSPHAT 1. Tn khc, ch s 2. nh ngha 3. Cm quan 4. Chc nng INS 452iv MTDI=70mg/kg th trng tnh cho phospho t cc ngun thc phm Hn hp d th ca cc mui calci ca acid polyphosphoric c cng thc chung l Hn+2PnOn+1. Tinh th hoc bt khng mi, khng mu. Cht nh ha, Cht gi m, cht to phc kim loi, cht to kt cu.

5. Yu cu k thut 5.1. nh tnh tan Thng khng tan hon ton trong nc, tan trong mi trng acid. Phosphat Phi c phn ng c trng ca phosphat. Calci Phi c phn ng c trng ca calci (Dch th chun b nh trong phn th phosphat) 5.2. tinh khit Gim khi lng Khng c qu 2%. Sy ti 105o trong 4 gi sau nung ti 550o trong 30 pht. khi nung Phosphat vng Khng c qu 8% tnh theo hm lng P2O5. Fluorid Khng c qu 10,0 mg/kg. Arsen Khng c qu 3,0 mg/kg. Ch Khng c qu 4,0 mg/kg. 5.3. Hm lng Khng c thp hn 50,0% v khng c qu 71,0% P2O5 P2O5 tnh theo ch phm sau khi nung. 6. Phng php th 6.1. nh tnh Phosphat Cn 0,5g mu, trn vi 10ml acid nitric v 50ml nc. un si trong 30 pht v lm mt. Dung dch ny c s dng lm dch th. 6.2. tinh khit Arsen - Th theo hng dn ti JECFA monograph 1 - Vol.4 - phng php II,.

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Ch

6.3. nh lng

- Ha tan 1 g mu th trong 15 ml dung dch acid hydrocloric long (TS), thm 20 ml nc, dung dch ny c s dng lm dch th). - 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. Cn khong 300 mg mu th, chnh xc n mg, ha tan trong 15 ml acid nitric v 30 ml nc ct. un si trong khong 30 pht, pha long vi nc n 100 ml. un nng n 60oC, thm lng d dung dch amoni molybdat (TS) v un 50oC trong 30 pht. Lc v ra ta bng acid nitric long (dung dch 1/36), tip theo bng dung dch kali nitrat (1/100) n khi dch ra khng cn acid vi qu tm. Ha tan ta trong 50 ml dung dch natri hydroxyd 1N, thm dung dch phenolphtalein (TS) v chun lng natri hydroxyd d bng acid sulfuric 1N. Mi ml dung dch natri hydroxyd 1 N tng ng vi 3,086mg P2O5.

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Ph lc 14 YU CU K THUT V PHNG PHP TH I VI CC MUI CA CC ACID BO Salts of fatty acids INS 470 ADI khng gii hn 2. nh ngha Cc sn phm ny cha mui calci, kali v natri ca cc acid myristic, oleic, palmitic, stearic hoc hn hp cc acid ny thu c t m v du n. Sn phm thng mi cn c phn loi da trn: - Ch s x phng ha; - im ng c ca cc acid bo thu c t mu th; - Ch s iod; - Cn cn li sau khi nung bao gm c hm lng cc cation; - m. 3. Cm quan Tinh th rn hoc na rn, bng, mu trng hoc vng nht, hoc bt mu trng hoc trng vng. 4. Chc nng Cht chng ng vn, Cht nh ha. 5. Yu cu k thut 5.1. nh tnh tan Cc mui kali v natri tan trong nc v ethanol, cc mui calci khng tan trong nc, ethanol v ether. Cation Phi c phn ng c trng ca cation. Thnh phn Phi c phn ng c trng ca thnh phn acid bo acid bo 5.2. tinh khit Cc acid bo t Khng c qu 3% do Cc cht khng Khng c qu 2 %. x phng ha Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng Khng thp hn 95 % tng cc mui acid bo, tnh theo khi lng kh. 6. Phng php th 6.1. nh tnh 1. Tn khc, ch s

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Cation

Thnh phn acid bo

un nng 1 g mu th vi hn hp gm 25 ml nc v 5 ml acid hydrocloric. Cc acid bo to thnh ni trn b mt thnh mt lp rn hoc du, tan trong hexan. Sau khi lm ngui, gn ly lp nc v cho bay hi n kh. ha tan cht rn cn li trong nc v th cc cation thch hp. S dng phng php nh lng nh tnh ring cc acid bo c trong thnh phn ca mu. Cc acid bo chim lng ln nht phi ph hp vi cc acid c a ra trn nhn sn phm.

6.2. tinh khit Cc acid bo t Xc nh cc acid bo t do theo hng dn trong phn Phng do php xc nh Acid bo t do. Tnh hm lng acid bo t do, s dng h s tng ng (e) tng ng vi 1/10 khi lng phn t ca mui. Cc cht khng Cc cht khng x phng ha l ton b lng sn phm c mt trong cc cht bo sau khi c x phng ha vi cc hydroxyd x phng ha kim loi kim v chit bng mt dung mi nht nh, vn khng bay hi trong cc iu kin xc nh ca php th. Cc cht ny bao gm cc lipid c ngun gc t nhin nh cc sterol, cc ru no mch di hn, cc phm mu v cc hydrocarbon cng nh bt k mt tp cht khng bay hi no 1030C c mt trong sn phm. Cn khong 5 g mu th (chnh xc n 0,01g) c trn u vo bnh cu y trn 250 ml. Thm 50 ml dung dch kali hydroxyd ~ 0,5 N v thm mt t bt. Gn bnh vo sinh hn hi lu, un si nh trong 1 gi. Ngng un. Thm 100 ml nc ct qua u sinh hn v lc. Sau khi lm mt, chuyn dung dch vo phu tch. Trng ra bnh v bt vi ln bng diethyl ether (dng khong 100 ml) v dch ra vo phu tch. ng nt v lc mnh trong 1 pht, u n x p sut bng cch lt ngc phu tch v m kha vi. yn phu tch cho n khi 2 pha hon ton tch lp. Sau rt ly cng trit cng tt dung dch x phng vo phu tch th 2. Chit dung dch x phng trong ethanol thm hai ln na, mi ln vi 100 ml diethyl ether theo cch trn. Gp 3 dch chit ether vo 1 phu tch c sn 40 ml nc. Nh nhng quay trn phu tch cha dch chit v 40 ml nc. Nu lc qu mnh trong giai on ny, nh tng c th hnh thnh. hn hp tch lp hon ton v rt b lp nc di. Ra lp ether hai ln na vi 40 ml nc, lc u v loi b lp

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nc di sau khi hn hp phn lp. Vi mi dung dch ra rt ra n khi ch cn khong 2 ml, sau quay trn phu tch quanh trc, i vi pht, rt b phn tch ra, ng kha vi khi ether bt u chy qua l ca kha vi. Ra dung dch ether nhiu ln bng 40 ml dung dch kali hydroxyd, 40 ml nc, ra thm mt ln bng 40 ml dung dch kali hydroxyd, sau ra t nht 2 ln na bng 40 ml nc. Lin tc ra bng nc cho n khi dung dch ht mu hng khi th bng ch th l dung dch phenophthalein. Chuyn ton lng tng t mt dung dch ether qua cung ca phu tch vo bnh cu kh v cn b khi lng chnh xc n 0,0001 g. Lm bay hi dung mi bng cch ct trong b cch thy. Thm 5 ml aceton v loi b hon ton dung mi bay hi bng cch thi khng kh nh, nghing bnh cu khi quay trong b cch thy. Sy kh phn cn cn li nhit 1032C trong 15 pht, t bnh cu theo phng gn nh nm ngang. Lm ngui trong bnh ht m v cn (chnh xc n 0,0001 g). Lp li qu trnh sy trong mi 15 pht lin tip cho n khi chnh lch gia 2 ln cn lin tip nh hn 0,0015g. Ch : Nu khng thu c khi lng khng i sau 3 ln sy kh, cc cht khng x phng ha c th b nhim bn. Sau khi cn, ha tan phn cn trong 4 ml diethyl ether, thm 20 ml ethanol c trung ha (dung dch c mu hng khi thm ch th l dung dch phenolphthalein(TS). Chun bng dung dch kali hydroxyd trong ethanol 0,1 N chun ha (pha dung dch kali hydroxyd trong ethanol c nng gn ng 1 N bng cch ha tan 60 g kali hydroxyd trong 50 ml nc v pha long thnh 1 L bng ethanol; pha long dung dch ny 1:10 bng ethanol) n khi dung dch c mu pht hng. Hiu chun khi lng cn cn li bng cch tr i hm lng acid t do. Tnh hm lng cht khng x phng ha, theo % (kl/kl) bng cng thc sau:

trong : m = khi lng (g), ca phn mu th, m1 = khi lng (g), ca cn cn li. V = th tch dung dch kali hydroxyd chun ha nng dng chun .

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T = Nng chnh xc ca dung dch kali hydroxyd dng chun . 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. 6.2. nh lng Nguyn tc: X phng ha cc mui v ester ha cc acid bo bng methanol, c mt bo trifluorid, methanol kim. Tin hnh sc k lng kh cc ester methyl ca acid bo. Phn A - Chun b cc ester methyl ca acid bo Thit b - Bnh y trn c nhm 50 v 100 ml. - Sinh hn hi lu, chiu di hiu dng 20 to 30 cm, c khp ni nhm tng thch vi bnh cu. - Cc phu tch 250 ml. - ng ng vo dn kh nitrogen. - ng nghim c nt thy tinh nhm. - Pipet chia , c th tch t nht l 10 ml, c gn qu bp cao su, hoc pipet t ng. Thuc th - Heptan, dng cho sc k (Ch 2 v 4) - Du nh c ct li (nhit si t 40-60), ch s brom < 1, khng c cn, hoc hexan (Ch 2) - Natri sulfat khan - Dung dch Natri hydroxyd trong methanol ~ 0.5 N: ha tan 2g natri hydroxyd trong 100 ml methanol cha khng qu 0.5% (kl/kl) nc. Nu gi dung dch tng i lu c th to thnh mt lng nh kt ta mu trng ca natri. iu ny khng nh hng n kt qu iu ch methyl ester. - Dung dch bo trifluorid trong methanol, 12 n 25% (kl/kl) . Ch phm thng mi sn c l dung dch 14 v 50% (ch 2) Cnh bo: Bo trifluorid c. V vy th nghim vin khng nn pha dung dch bo trifluorid trong methanol t methanol v boron trifluorid. (ch 3).

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- Dung dch bo ha natri clorid trong nc. - Methyl , dung dch 1 g/l trong ethanol 60% (tt/tt) - Nitrogen, c hm lng oxy < 5 mg/kg Tin hnh Do bo trifluorid c, cc th nghim sau tt nht nn tin hnh trong t ht. Cc dng c thy tinh phi c ra bng nc ngay sau khi s dng. Sy kh mu nhit 105C n khi lng khng i, c 2 gi th kim tra khi lng 1 ln. Cn khong 350mg (chnh xc n mg) mu sy kh. C th ly lng mu ln hoc nh hn 350 mg, tuy nhin, th tch ca bnh cu v lng thuc th s dng phi ph hp vi s liu trong bng sau: Khi lng mu (mg) 100-250 250-500 500-700 750-1000 Th tch bnh (ml) 50 50 100 100 NaOH 0.5 N (ml) 4 6 8 10 Dung dch BF3 trong methanol (ml) 5 7 9 12

Cho lng acid bo nh vo bnh cu ph hp. Thm vi lng ph hp dung dch bo trifluorid trong methanol. un si trong vng 2 pht. Thm 2 n 5 ml heptan (ch 4) (lng chnh xc khng nh hng n phn ng) qua u trn ca sinh hn vo hn hp ang si v tip tc un si trong 1 pht. Ngt ngun nhit, ly sinh hn ra. Thm lng nh dung dch natri clorid bo ha v lc nh bnh bng cch quay trn bnh vi ln. Thm dung dch natri clorid bo ha vo bnh cu sao cho mc cht lng ngang c bnh. yn cho hn hp tch lp v chuyn khong 1 ml lp trn (lp heptan) vo ng nghim c nhm v thm vo mt lng nh natri sulfat khan loi ht nc. Nu lng mu ly l 350 mg, dung dch ny cha khong 7-17% methyl ester v c th bm trc tip vo ct sc k kh-lng. Nu khng th pha long dung dch bng heptan t c nng methyl ester l 5-10% (ch 5). thu hi ton b lng ester kh, chuyn dung dch mui v lp heptan vo phu tch. Tch ring cc lp. Chit phn dung dch

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mui 2 ln bng 50 ml du nh. Gp dch chit du nh thu c vo phn dung dch heptan, ra bng 20 ml nc n khi ht acid (th bng ch th methyl). Lm kh bng natri sulfat khan, lc v cho bay hi dung mi trong b cch thy v thi kh nitrogen (ch 6 v 7). Vi cc mu c khi lng nh hn 500 mg phi gim th tch dung mi v nc s dng theo t l tng ng. Bn cnh phng php trn cn c mt s phng php khc khng s dng n bo trifluorid. Nhn chung c th thay cc thuc th methyl ha, dung dch natri hydroxyd 0,5N trong methanol v dung dch bo trifluorid 12 - 25 %, trong methanol bng cc cht sau: - Kali hydroxyd 1 N trong methanol, (phn ng vi cht bo khi c mt lng d methanol c hm lng nc nh); - Dung dch natri methylat (iu ch bng cch ha tan 1 g natri kim loi trong 100 ml methanol c hm lng nc nh). Ch 1. Nu cc cht khng x phng ha gy cn tr, pha long dung dch x phng ha vi nc v loi b cc cht khng x phng ha bng cch chit vi diethyl ether hoc hexan. Acid ha dung dch nc x phng v tch ly cc acid bo. iu ch cc methyl ester t cc acid bo theo hng dn. 2. Khi o sc k kh lng ca cc methyl ester, mt s thuc th, c bit l bo trifluorid trong methanol c th to nn cc peak ngu nhin trn sc (trong vng cc ester c mch C20-C22 nu dng bo trifluorid trong methanol). Do vy cn phi kim tra li mi t thuc th mi bng cch iu ch cc methyl ester ca acid oleic tinh khit, v o sc k. Nu xut hin peak ngu nhin, phi b khng dng thuc th. Cc thuc th khc nhau khng c php cho peak gy nhiu n cc peak ca cc methyl ester ca cc acid bo trong qu trnh chy sc k. Dung dch bo trifluorid trong methanol phi c bo qun trong t lnh. 3. Nu bt buc phi iu ch dung dch bo trifluorid t bo trifluorid kh, nn dng phng php sau: Cn khi lng ca bnh cu 2 L cha 1 L methanol. Lm lnh trong b . Vn gi nguyn bnh trong b , sc kh BF3 t chai kh (cylinder) qua ng dn thy tinh vo dung dch methanol cho n khi hp th c 125 g BF3. Tin hnh th nghim trong t ht. Phi cho BF3 chy qua ng thy tinh trc khi t ng vo v rt ng ra khi methanol trnh trng hp cht lng b ht vo h thng van

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ca chai kh. Khng c cho kh chy nhanh qu c hi kh trng ra khi bnh. Thuc th pha theo cch ny bn trong 2 nm. 4. Nu cc acid bo cha 20 hoc nhiu hn 20 nguyn t carbon khng c mt, nn thay heptan bng hexan (hn hp ca cc ng phn C7 tinh khit c th c kim tra bng sc k kh lng) 5. Nu khng c lng mu nh yu cu, c th ly 10 mg, thm ch t hn, min l lng thuc th v kch c ca cc dng c thy tinh c gim theo t l tng ng. 6. Cc dung dch methyl ester nn c phn tch cng sm cng tt. Nu cn, bo qun dung dch heptan cha cc methyl ester trong mi trng kh tr trong t lnh. Nu cn phi bo qun trong thi gian di, nn bo v cc methyl ester khi s oxy ha bng cch thm vo dung dch cc cht chng oxy ha nng khng nh hng n kt qu phn tch, v d nh 0,05 g/L EHT (2,6-di-tertbutyl-4-methyl phenol). Trong trng hp cn thit, c th bo qun cc methyl ester kh sch dung mi trong kh tr trong t lnh trong 24 gi, hoc c th bo qun lu hn nu gi ng lnh su trong ng kn trong chn khng. 7. Mt phn cc methyl ester d bay hi nht c th b mt nu thi gian bay hi dung mi ko di, hoc nu dng kh nitrogen qu mnh, o ph hng ngoi, phi loi b dung mi cng trit cng tt. o sc k kh lng, loi b hon ton dung mi. Phn B - Sc k kh lng ca cc ester methyl acid bo Thit b Thit b thng thng dng cho sc k kh lng, dng ct nhi v detector ion ha ngn la (ch 1). Bt k mt thit b no cho hiu qu v phn gii cao vi cht bo cn xc nh u ph hp. Sc k kh lng H thng bm mu: H thng bm mu phi c khng gian cht nh nht c th. Nu c, phi nng c nhit cao hn nhit ct t 20 n 50C. L: L phi cung cp cho ct nhit t nht l 220o v phi duy tr c nhit cn thit trong khong 1C. Nu dng chng trnh t nhit , nn dng thit b c ct i.

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Ct nhi: - Ct: Ct phi c lm t lai vt liu tr vi cht cn phn tch: thy tinh hoc nu khng th dng thp khng r (ch 2). Chiu di: 1 n 3 m, nn dng ct tng i ngn nu c cc acid mch di (C20+). xc nh cc acid bo C4 v C6, nn dng ct 2 m; ng knh trong t 2 n 4 mm. Nhi ct - Cht mang: Diatomid ra acid v c silan ha, hoc cc cht mang tr ph hp khc c khong bin thin ng knh hp (25m) trong khong ng knh c ht t 125-200 m, c ht trung bnh phi tng quan vi ng knh trong v chiu di ct. - Pha tnh: Dng polyester ca mt cht lng phn cc (v d diethylen glycol polysuccinat, butandiol polysuccinat, ethylen glycol polyadipat ...) hoc bt k cht lng no khc (v d cyanosilicon ...) tha mn cc iu kin di y. Pha tnh phi chim 5 n 20 % khi lng nhi. C th dng mt pha tnh khng phn cc, ty theo tng php tch c th. - Luyn ct mi ch to: Tch ct ra khi detector. Nu c th, t t nung l n 185C v dn dng kh tr qua ct vi tc 20 - 60 ml/pht trong t nht l 16 gi ti nhit ny, v thm 2 gi na ti 195C. Detector: Cc vn hnh di y tng ng vi detector ion ha ngn la (ch 1) Syring: Syring, c dung tch ti a l 10 l, chia n 0.1 l. My ghi: Nu s dng ng ghi tnh thnh phn hn hp phn tch, phi dng mt my ghi in t c chnh xc cao. My ghi phi tng thch vi thit b o. Cc c trng ca my ghi phi nh sau: - Tc p ng di 1,5 giy, tt nht l di 1 giy (tc p ng l thi gian cn bt ghi chy t 0 n 90 % theo 100 % tn hiu) - rng ca giy: ti thiu l 25 cm - Tc giy: 25-100 cm/gi. B tch phn Integrator hoc my tnh (ty chn) C th tnh mt cch nhanh v chnh xc nu dng b tch phn in t hoc my tnh. Trong trng hp ny phi to c tn hiu tuyn tnh vi nhy ph hp, v phn hiu chnh cho s lch ng nn phi tha ng.

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Ha cht - Kh mang: Kh tr (nitrogen, heli, argon ...) kh, v cha t hn 10 mg/kg oxy. - Kh ph tr: Hydrogen (tinh khit ti thiu 99.9%) khng c cc tp cht hu c, khng kh hoc oxygen. - Cc cht chun so snh: Hn hp ca cc methyl ester, hoc cc methyl ester ca mt cht du bit trc thnh phn, tt nht l tng t vi cht bo cn phn tch. Tin hnh iu kin th Xc nh cc iu kin vn hnh ti u: Cc gi tr cho trong bng 1 v 2 cho di y v nguyn tc c th cho cc kt qu mong mun: Bng 1 ng knh trong ca ct Kh mang cung cp 2 mm 15-25 ml/pht 3 mm 20-40 ml/pht 4 mm 40-60 ml/pht Bng 2 Nng ca pha tnh Nhit 5% 175o 10% 180o 15% 180o 20% 185o Nu c, injector phi nhit khong 200oC v detector phi nhit bng hoc cao hn nhit ct. V nguyn tc dng kh hydrogen cung cp cho detector ion ha ngn la phi c tc bng mt na tc kh mang, v tc kh oxygen phi bng khong 5 n 10 ln tc kh hydrogen. Xc nh hiu sut v phn gii (ty chn) Tin hnh phn tch mu chun methyl stearat. La chn c mu, nhit ct tch v tc kh mang sao cho peak ca methyl stearat xut hin 15 pht sau peak dung mi v cao khong 3/4 tn hiu ton thang. - Phn tch: Dng mu kim tra l 0,1 n 2 l dung dch heptan ca methyl esters,

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pha theo hng dn trong phn A. Trong trng hp khng c cc ester trong dung dch, chun b dung dch c nng khong 10% trong heptan v bm 0,1 n 1 l dung dch ny. Cc iu kin tin hnh c cho di y. Tuy nhin, c th lm vic vi nhit ct thp hn nu xc nh cc acid c mch carbon di C12 hoc nhit cao hn nu xc nh cc acid bo trn C20. i khi c th s dng chng trnh t nhit cho c 2 trng hp trn. Nu mu c cha cc methyl ester ca cc acid bo di C12, phi bm mu nhit 100C (hoc nhit t 50 60C nu c acid butyric) v ngay lp tc tng nhit vi tc tng nhit t 4 - 8 C/pht cho n nhit ti u. Trong mt s trng hp c th gp c 2 quy trnh li: sau khi tng nhit, tip tc ra gii ti nhit khng i cho n khi tt c cc hp phn c ra gii ra. Nu thit b khng vn hnh vi chng trnh nhit , lm vic ti 2 nhit c nh trong khong t 100 n 195C. Biu th kt qu Phn tch nh tnh Phn tch hn hp cht chun so snh c thnh phn bit trong cc iu kin s dng o mu th. o khong cch lu (hoc thi gian lu) cho cc ester bo tng ng. Dng giy bn logarit, v th biu din s ph thuc ca logarit khong cch lu (hoc thi gian lu) vo s nguyn t carbon ca cc acid; trong iu kin ng nhit th ca cc ester mch thng c cng khng bo Ha (no) phi l cc ng thng. gn nh song song vi nhau. Nhn dng cc peak ca mu th t cc th ny, nu cn th dng php ni suy. Phi trnh cc iu kin o lm xut hin cc peak b che ph (masked peaks), c ngha l phn gii khng ln tch c 2 hp phn. Phn tch nh lng Xc nh thnh phn Tr cc trng hp c bit, s dng phng php chun ha din tch, c ngha l gi thit rng ton b cc hp phn u c biu din trn sc , sao cho tng din tch cc peak biu din 100 % cc hp phn (tng ra gii). Bng mt qui trnh chun ha ph hp (dng hn hp chun so snh hoc dng cht chun ni), xc nh tng khi lng ca cc acid bo trong mu kh. Tnh hm lng mui ca cc acid

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bo vi cc cation c trng trong mu. Tng hm lng Mui Acid Bo khng c ln hn 95% khi lng mu kh. Ngoi ra, nu c cc ch tiu cho hm lng ca mi acid bo ghi trn nhn sn phm, mu phi tha mn c cc ch tiu ny. Ch 1. C th s dng my sc k kh - lng dng thit b o dn nhit ca kh (catharometer) (lm vic trn nguyn tc o s thay i dn nhit). Cc iu kin o khi c thay i nh sau: Ct - Chiu di: 2 n 4 m - ng knh trong: 4 mm - Cht mang: ng knh ht trong khong 160 n 200 m - Pha tnh: 15 n 25% Kh mang: heli, nu khng th dng hydrogen, c hm lng oxygen cng nh cng tt. Khng dng kh ph tr - Tc kh: thng trong khong t 60 n 80 ml/pht Nhit Injector: cao hn t 40o n 60oC so vi nhit ct Ct: 180o to 200oC Phn tch nh lng: phi hiu chnh theo cc h s thu c khi phn tch hn hp so snh ca cc ester c thnh phn bit trong cng iu kin o vi mu th. 2. Nu trong mu c cc thnh phn cha nhiu hn 3 ni i, chng c th b phn hy trn ct thp khng r.

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Ph lc 15 YU CU K THUT V PHNG PHP TH I VI ESTER CA SUCROSE VI CC ACID BO Sucrose fatty acid esters; INS 473 2. nh ngha Mono, di, tri ester ca sucrose vi cc acid bo thc phm, c iu ch t sucrose v methyl, ethyl ester ca cc acid bo thc phm bng cch ester ha vi s c mt ca cht xc tc, hoc tch t sucroglycerid. Ch cc dung mi sau c th c s dng sn xut: dimethylformamid, dimethyl sulfoxid, ethyl acetat, isopropanol, propylen glycol, isobutanol v methyl ethyl keton. 3. Cm quan Dng gel cng, dng cht c mm hoc dng bt c mu trng n trng hi xm 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan t tan trong nc, tan trong ethanol. Acid bo Phi c phn ng c trng ca acid bo. ng Phi c phn ng c trng ca ng. 5.2. tinh khit Tro sulfat Khng c qu 2% Th 1 g mu (Phng php I) Ch s acid Khng c qu 6 Sucrose t do Khng c qu 5% (m t trong phn Phng php th) Dimethylformamid Dimethyl sulfoxid Ethyl acetat, isopropanol v propylen glycol Isobutanol Methanol Methyl ethyl keton Khng c qu 1,0 mg/kg (m t trong phn Phng php th) Khng c qu 2,0 mg/kg (m t trong phn Phng php th) Khng c qu 350,0 mg/kg, dng n cht hoc hp cht (m t trong phn Phng php th) Khng c qu 10,0 mg/kg (m t trong phn Phng php th) Khng c qu 10,0 mg/kg (m t trong phn Phng php th) Khng c qu 10,0 mg/kg (m t trong phn Phng php th 1. Tn khc, ch s

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Ch Khng c qu 2,0 mg/kg. 5.3. Hm Khng c nh hn 80,0% ester sucrose lng 6. Phng php th 6.1. nh tnh Acid bo Cho 1 ml ethanol vo 0,1 g mu, lm m v ha tan, thm 5 ml dung dch H2SO4 long TS, gia nhit trong cch thy trong 30 pht v lm ngui. Cht c hoc du mu trng vng c hnh thnh, khng c mi ca acid isobutyric v c ha tan khi thm 3 ml diethyl ether. S dng lp dung dch nc tch khi t diethyl ether cho Php th ng. ng Cho vo 2 ml lp dung dch nc tch khi diethyl ether trong php th acid bo, 1 ml anthron TS vo trong ng th, cho cn thn theo thnh ng nghim; b mt ranh gii gia hai lp chuyn thnh mu xanh da tri hoc xanh l cy. 6.2. tinh khit Sucrose t do Xc nh bng sc k kh lng (Quyn 4) Dung dch chun: Chun b dung dch gc cha 5 mg/ml sucrose trong N, N- dimethylformamid. Chun b mt lot cc dung dch chun cha 0,5; 1,25 v 2,5 mg/ml sucrose bng cch pha long dung dch gc vi pyridin. Dung dch ni chun: Cn chnh xc 0,25 g octacosan cho vo bnh nh mc dung tch 50 ml, thm vo 25 ml tetrahydrofuran ha tan octacosan, v nh mc n th tch 50 ml bng pyridin. Cc iu kin sc k: Ct: 2% Dexsil 300GC trn Uniport HP 80/100 mesh (t phn cc, 2,1 m x 3.2 mm ng knh trong) hoc tng ng Kh mang: nitrogen Detector: FID Nhit : - Bung bm mu: 2800C - Ct: gi 1600C trong 1 pht sau tng t 160 n 3000C vi tc 150C/pht, gi trong 60 pht 3000C. - Detector: 3200C Thi gian lu ln lt ca sucrose v octacosan o c vi cc iu kin trn xp x 8,2 v 9,8 pht.

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Cch tin hnh: Cn chnh xc 20 - 50 mg mu cho vo ng ly tm, thm 1 ml dung dch ni chun, 1 ml pyridin, 0,4 ml N, O-bis (trimethylsilyl) acetamid (BSA) v 0,2 ml trimethylchlorosilan (TMCS). Sau khi y np ng, lc v yn trong 5 pht nhit phng. Bm 1l vo thit b sc k kh lng. ng chun: Chun b dung dch chun silylat theo cch tin hnh nh nhau s dng 1 ml mi loi dung dch chun thay th mu v pyridin. V ng chun trn th lng sucrose (mg) trong 1 ml dung dch chun (biu th trn trc X) so vi t l din tch pic ca sucrose/ni chun (biu th trn trc Y). o din tch pic i vi sucrose v ni chun. Tnh t l din tch cc pic ca chng, v tnh c hm lng sucrose trong mu da vo ng chun. Lng sucrose pht hin c (mg) % sucrose = ----------------------------------------------------- x 100 Khi lng mu (mg) Dimethylformamid Xc nh bng sc k kh lng Dung dch chun: Chun b dung dch gc cha 1 mg/ml dimethylformamid trong tetrahydrofuran. Chun b mt lot cc dung dch chun cha 0,05; 0,1 v 0,2 g/ml dimethylformamid bng cch pha long dung dch gc vi tetrahydrofuran. Cc iu kin sc k: Ct: polyethylen glycol (di 30 m x 0,32 mm ng knh trong vi dy lp phim mng 0,5 m) Kh mang: heli p sut: 150 kPa (p sut khng i) Detector: detector nitrogen phosphor (NPD) [(tn khc: Detector nhit ion ngn la (FTD)] Nhit : - Bung bm mu: 1800C - Ct: gi 400C trong 2 pht sau tng t 40 n 1600C vi tc 200C/pht, gi trong 2 pht 1600C. - Detector: 3250C Phng php tim mu: Tim khng chia dng 1,0 l mu bng bm t ng, sau bt u lm sch sau 1 pht.

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Thi gian lu ca dimethylformamid o c vi cc iu kin trn xp x 6,4 pht. Cch tin hnh: Cn chnh xc 2 g mu cho vo bnh nh mc dung tch 20 ml, thm vo 10 ml tetrahydrofuran ha tan mu, thm tetrahydrofuran vo bnh n th tch 20 ml v lc u dung dch. Bm 1,0 l dung dch mu vo thit b sc k. ng chun: Chun b hng ngy bng cch bm 1,0 l mi dung dch chun vo thit b sc k. Tnh nng dimethylformamid (CDFA) theo cng thc sau: Trong : C: Nng dimethylformamid pht hin c (g/ml) W: Khi lng mu (g) Lu : Detector nitrogen phosphor khng nhy vi cc thnh phn khng cha N hoc P. V th, ct mao qun c th tc vi cc hp cht bay hi thp, mc d ng nn ca sc k n nh. Do , ct phi c luyn thng xuyn. Luyn qua m (dng kh mang theo hng ngc 1800C) c thc hin sau khong mi t 15 mu. Xc nh bng sc k kh lng Dung dch chun: Chun b dung dch gc 0,25 mg/ml dimethyl sulfoxid trong tetrahydrofuran. Chun b mt lot cc dung dch chun cha 0,5; 1 v 5 g/ml dimethyl sulfoxid bng cch pha long dung dch gc vi tetrahydrofuran. Cc iu kin sc k: Ct: 10% PEG 20M v 3% KOH trn Gas Chrom Z (chiu di 2 m x 3 mm ng knh trong). Tng nhit l ti 1800C vi tc 100C/pht v n nh 24 - 48 gi vi iu kin tc dng kh nitrogen 30 - 40 ml/pht luyn ct. Kh mang: nitrogen Tc dng: 50 ml/pht Detector: detector trc quang ngn la (s dng b lc lu hunh 394 nm)

Dimethyl sulfoxid

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Nhit : - Bung bm mu: 2100C - Ct: 1600C. Thi gian lu ca dimethyl sulfoxid o c vi cc iu kin trn xp x 3,4 pht. Cch tin hnh: Cn chnh xc 5 g mu cho vo bnh nh mc dung tch 25 ml, thm vo 10 ml tetrahydrofuran ha tan mu, thm tetrahydrofuran vo bnh n th tch 20 ml v lc u dung dch. Bm 3 l dung dch mu vo thit b sc k. ng chun: Chun b hng ngy bng cch bm 3 l mi dung dch chun vo thit b sc k. Tnh nng dimethyl sulfoxid (CDMSO) theo cng thc sau: Trong : C: Nng dimethyl sulfoxid pht hin c (g/ml) W: Khi lng mu (g) Propylen glycol Xc nh bng sc k kh lng Dung dch ni chun: Chun b dung dch 500 g/ml ca ethylen glycol trong pyridin Dung dch chun: Chun b dung dch 50 g/ml ca propylen glycol trong pyridin Cc iu kin sc k: Ct: Polydimethylsiloxan (chiu di 30 m x 0,32 mm ng knh trong vi dy lp phim mng 0,25 m) Kh mang: heli Tc dng: 1,5 ml/pht (dng khng i) Detector: FID Nhit : - Bung bm mu: 2300C - Ct: gi 600C trong 5 pht sau tng t 60 n 2500C vi tc 200C/pht, gi trong 5 pht 2500C. - Detector: 2500C Thi gian lu ln lt ca dn xut ethylen glycol v dn xut propylen glycol xp x 7,7 pht v 7,9 pht.

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Cch tin hnh: Cn chnh xc 1 g mu cho vo bnh nh mc dung tch 10 ml, thm vo 100 l dung dch ni chun. ha tan v nh mc n th tch 10 ml bng pyridin. Ly 0,5 ml dung dch mu cho vo ng li tm, v thm 0,25 ml 1,1,1,3,3,3 - hexamethyldisilazan (HMDS) v 0,1 ml trimethylchlorosilan (TMCS). Sau khi y cht np ng, lc mnh, yn trong 30 pht nhit phng, sau ly tm. Bm 1 l dung dch ni trn mt ng sau ly tm vo thit b sc k. ng chun: Tin hnh sc k nh trn, trong dng 0,05; 0,2; 0,5 v 1ml dung dch chun thay th cho dung dch mu th. Tnh nng propylen glycol (CPG) theo cng thc sau: Trong : C: Nng propylen glycol pht hin c (g/ml) W: Khi lng mu (g) Lu : Nht thit phi lm sch cng bm mu v luyn ct 3000C sau mi t 20 mu phn tch, bi ct b nhim bn. Methanol, isopropanol, isobutanol, ethyl acetat v methyl ethyl keton Xc nh bng sc k kh c bung ly mu khng gian hi. Dung dch chun: Chun b dung dch chun A cha 4.000 mg/l mi loi methanol, isopropanol, isobutanol, ethyl acetat v methyl ethyl keton bng cch cn chnh xc 0,2 g mi dung mi v cho vo bnh nh mc dung tch 50 ml cha khong 20 ml nc, sau thm nc vo n th tch 50 ml. Chun b cc dung dch cha 2.000 mg/l (dung dch chun B) v 1.000 mg/l (dung dch chun C) bng cch pha long dung dch chun A. Cch tin hnh: Cn chnh xc 4 g mu cho vo 4 l (mi l 1 g mu). Cho 5 l nc vo l th nht, cho ln lt vo l th hai, th ba, th bn dung dch chun A, B v C; y nhanh septum. (Nng ca mi dung mi sau khi thm 5 l dung dch chun A, B, C vo 1 g mu tng ng ln lt bng 20, 10 v 5 mg/kg mu). l mu trong bung ly mu khng gian hi v phn tch vi cc iu kin sau:

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Ct: 100% Polydimethylsiloxan (chiu di 30 m x 0,53 mm ng knh trong vi dy mng phim mng 1,5 m), v d: DB - 1 c sn xut bi Cng ty TNHH J & W) Kh mang: nitrogen Tc dng: 3,5 ml/pht Detector: FID Nhit : - Bung bm mu: 1100C - Ct: 400C. - Detector: 1100C Bung ly mu khng gian hi: - Nhit cch nhit mu: 800C - Thi k cch nhit mu: 40 pht - Nhit syringe: 850C - Bm mu dng kh: 1,0 ml Kt qu: th biu din mi tng quan gia lng thm vo vi din tch pic i vi mi dung mi s dng cc kt qu phn tch. Mi tng quan phi tuyn tnh (R2 > 0,99). Ngoi suy v xc nh x phn b chn, wi, v tnh nng dung mi Ci (mg/kg) trong mu: Trong : wi: Phn b chn ca ng thng tng quan s dng phng php thm chun (g) W: Khi lng mu (g) 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 Xc nh bng HPLC s dng cc iu kin sau: Cch tin hnh: Cn chnh xc 250 mg mu v cho vo bnh nh mc dung tch 50 ml. Pha ti th tch 50 ml bng tetrahydrofuran v lc u. Lc qua mng lc 0,5 m. Bm 100 l mu vo thit b sc k n nh trc.

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Cc iu kin sc k: Ct: Styren-divinylbenzen copolymer i vi sc k thm thu qua gel (TSK-GEL G2000 (Tosoh) hoc tng ng) Pha ng: tetrahydrofuran dng cho HPLC bi kh Tc dng: 0,7 ml/pht Detector: RI Nhit : - Ct: 380C. - Detector: 380C Ghi sc k trong khong 90 pht. Tnh % ester sucrose trong mu: % ester sucrose = 100 A/T Trong : A: Tng cc din tch pic i vi 3 thnh phn chnh, mono-, di- v tri-ester, ra gii ln lt khong 65, 68 v 73 pht. T: Tng din tch tt c cc pic ra gii trong khong 90 pht

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Ph lc 16 YU CU K THUT V PHNG PHP TH I VI DIOTYL NATRI SULFOSUCINAT 1. Tn khc, ch s 2. nh ngha Tn ha hc DSS, natri docusat, INS 480 ADI = 0 - 0,1 mg/kg th trng

Natri1,4-bis-(2-ethylhexyl)-sulfosuccinat; Mui natri ca acid ester sulfo-1,4-bis-(2-ethylhexyl) butanedioic; dioctyl natri sulfosuccinat S C.A.S. 577-11-7 Cng thc ha hc C20H37NaO7S Cng thc cu to

Khi lng phn t 3. Cm quan

444,56

Cht rn dng do, ging sp, mu trng c mi c trng ging octanol, nhng khng c mi ca cc dung mi khc. 4. Chc nng Cht nh ha, cht lm m 5. Yu cu k thut 5.1. nh tnh tan t tan trong nc, d tan trong ethanol v glycerol Ph hng ngoi Ph hng ngoi ca mu phn tn trong kali bromid tng ng vi ph ca mu chun i chiu (Chun i chiu USP dioctyl sulfosuccinate c cng ty United States Pharmacopoeia, 12601 Twinbrook Parkway, Rockville, Md 20852, USA) pH 5.8 - 6.9 (dung dch 1/100) 5.2. tinh khit Gim khi lng Khng c qu 2 % (105C trong 2 gi) khi lm kh trong ca Ha tan 25 g mu th vo 94 ml ethanol. Dung dch khng c vn c sau 24 gi. dung dch

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Trong khong t 15,5 % n 16,2 %. (th theo hng dn ti JECFA monograph 1 - Vol.4; Phng php I, cn 1 g mu) Bis-(2-ethylhexyl)- Khng c qu 0,4 %. maleat Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng Khng thp hn 98,5 % tnh theo khi lng kh. 6. Phng php th 6.1. tinh khit Bis-(2Thuc th ethylhexyl)- Cht in gii h tr: ha tan 21,2 g lithi perchlorat (LiClO4) maleat khan vo 175 ml nc trong cc 250 ml. iu chnh pH ca dung dch ny n 3,0 bng cch thm tng git acid acetic bng (khong 1 n 2 git l ), dng mt my o pH ph hp. Chuyn ton lng dung dch vo bnh nh mc 200 ml, pha long n vch bng nc, v trn u. - Dung dch chun: Chuyn 100 - 110 mg (cn chnh xc) bis(2-ethyl-hexyl)-maleat, (bis-(2-ethylhexyl)-maleat vi tinh khit ph hp c tn khc l OT-35 ca Cng ty American Cyanamid, Pine Chemicals Department, Pearl River, New York 10965, USA), vo bnh 100 ml. Ghi li khi lng chnh xc, chnh xc n 0,1 mg, Wa. Pha long n vch bng nc, trn u. - Dung dch mu th gc: Chuyn 12,5 g mu th, c cn chnh xc, vo cc 150 ml. Ghi li khi lng chnh xc, Ws, chnh xc n 10 mg. Thm 80-90 ml isopropanol, khuy bng a thy tinh n khi mu tan hon ton. Chuyn ton lng dung dch ny vo bnh nh mc 250 ml bng isopropanol. Pha long n vch bng isopropanol, trn u. - Dung dch th A: Dng pipet ly 50.0 ml "Dung dch mu th gc" v 20,0 ml "Dung dch in gii h tr " vo bnh nh mc 100 ml. Pha long vi isopropanol n khong di vch nh mc 15 mm, y nt, lc u v trong 2 pht. Pha long tip bng isopropanol n vch, y nt, lc u. Phi thu c dung dch tuyt i trong. - Dung dch th B: Dng pipet ly 50,0 ml "Dung dch mu th gc", 10,0 ml "Dung dch chun", v 20,0 ml Dung dch in gii h tr" vo bnh nh mc 100 ml, v tip tc qui trnh nh miu t trong phn Dung dch th A - Mu trng: Dng pipet ly 20,0 ml "Dung dch in gii h tr" vo bnh nh mc 100-ml, pha long n vch bng isopropanol, trn u.

Tro sulfat

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Tin hnh Ra bnh cc ph H-cell vi ln bng tng lng nh Dung dch th A, sau y mt na bnh bng dung dch ny, t mt mu giy thm ln ming bnh v cho dng kh nitrogen chy t t qua dung dch trong 15 pht (Ch : Trc ht phi bo ha nitrogen bng cch cho chy qua bnh sc ph hp c cha isopropanol). Sau 15 pht, hng dng nitrogen ln b mt dung dch v ly giy ra khi bnh. Vi mt cc ph k ph hp c hiu chun trc (nh cc ph k Metrohm Polacord E-261, hoc tng ng t mt th phn cc -1,3V. iu chnh nhy dng ti thang thp nht (nhy nht) m ti cc dao dng dng vn nm trong thang. Ghi li cc ph ti nhy ny, qut th trong khong t 0,9 n -1,5 V, dng in cc so snh l in cc calomel bo ha . Ghi li dao ng trung bnh A ti th -1,3 V, v dao ng trung bnh B, ti th -1,0 V, theo mm. (Ch : nu s dng cc ph k khng t ng, ghi li cc dao ng dng khuch tn trung bnh ca dung dch tng ng ti -1,3 V v -1,0V). Lp li ton b qu trnh vi dung dch th B, ghi li gi tr dao ng dng khuch tn trung bnh, D, ti -1,3 V v E, ti - 1,0 V. Lm tng t vi mu trng, ghi li gi tr dao ng dng khuch tn trung bnh, G, ti -1,3V v H, ti -1 V. Tnh ton: Trc ht tnh cc ga tr sau (theo mA): Trong C l cng dng khuch tn ca Dung dch th A v S1 l nhy dng khi o "Dung dch th A" Trong F l cng khuch tn ca Dung dch th B v S2 l nhy dng khi o "Dung dch th B" Trong I l cng dng khuch tn ca Mu trng v S3 l nhy dng khi o "Mu trng" Trong J l dng khuch tn gy ra do thm maleat t Dung dch chun vo Dung dch th B.

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Trong K dng khuch tn gy ra do maleat c trong Dung dch th A. Cui cng, tnh phn trm ca bis-(2-ethylhexyl)-maleat c trong mu ban u theo cng thc sau:

Ch

6.2. nh lng

- 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. Cc dung dch - Dung dch mu: Chuyn khong 3,8 g mu th, trc c sy kh 105C trong 2 gi v c cn chnh xc, vo bnh nh mc 500 ml, ha tan trong cloroform. Pha long n vch bng dung mi trn, trn u. - Dung dch tetra-n-butylamoni iodid: Chuyn 1,250 g tetra-nbutylamoni iodid vo bnh nh mc 500 ml, pha long n vch bng nc, trn u. - Dung dch mui: ha tan 100 g natri sulfat khan v 10 g natri carbonat vo mt lng nc va c c th tch cui cng l 1000 ml. Tin hnh Dng pipet ly 10,0 ml "Dung dch mu" vo bnh nh mc 250 ml, thm vo 40 ml cloroform, 50 ml "Dung dch mui", 10 git dunh dch xanh bromophenol (TS). Chun bng "Dung dch tetra-n-butylamoni iodid" n khi bt u xut hin mu xanh trong lp cloroform sau khi lc mnh dung dch. Tnh hm lng (%) C20H37NaO7S theo cng thc:

trong V = th tch (ml) ca dung dch tetra-n-butylamoni iodid dng chun . 444,6 = khi lng phn t ca dioctyl natri sulfosuccinat W = khi lng (g) ca mu th 369,4 = khi lng phn t ca tetra-n-butylamoni iodid.

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Ph lc 17 YU CU K THUT V PHNG PHP TH I VI STEARYL TARTRAT Stearyl palmityl tartrat; INS 483 2. nh ngha Sn phm ca qu trnh ester ha acid tartric vi cn stearylic thng mi, v c bn gm cn stearylic v palmitylic; gm thnh phn chnh l diester, vi lng nh monoester v cc nguyn liu ban u khng thay i. Tn ha hc Cc thnh phn chnh l distearyl tartrat, dipalmityl tartrat v stearylpalmityl tartrat. Cng thc Distearyl tartrat: C40H78O6 ha hc Dipalmityl tartrat: C36H70O6 Stearylpalmityl tartrat: C38H74O6 Cng thc cu to 1. Tn khc, ch s

Trong R l (CH2)17CH3 hoc (CH2)15CH3 Khi lng Distearyl tartrat: 655,06 phn t Dipalmityl tartrat: 598,95 Stearylpalmityl tartrat: 627 3. Cm quan Cht nhn c mu kem 4. Chc nng Cht nh ha, cht x l bt 5. Yu cu k thut 5.1. nh tnh tan Khng tan trong nc, khng tan trong ethanol lnh, tan trong ethanol nng. Nhit 67 - 770C nng chy Ch s hydroxyl 200 - 220 Tartrat Phi c phn ng c trng ca tartrat. 5.2. tinh khit Tro sulfat Khng c qu 0,5% (Th trn 2 g mu, phng php I ) Acid tartric Khng c nh hn 18% v khng c qu 35% (m t trong tng s phn Phng php th)

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Cht khng Khng c nh hn 77% v khng c qu 83% (m t trong x phng ha phn Phng php th) Ch s acid Khng c qu 6 (m t trong phn Phng php th) Ch Khng c qu 2,0 mg/kg. 5.3. Hm Khng c nh hn 90% hm lng ester tng s tng ng vi lng ch s ester trong khong 163 - 180. 6. Phng php th 6.1. tinh khit Acid tartric ng chun: tng s Cn 100 mg acid tartric tinh khit cho vo bnh nh mc dung tch 100 ml, ha tan trong khong 90 ml nc, thm nc vo cho n khi t th tch 100 ml v lc u. Chuyn cc phn tch bit 3, 4, 5, 6 ml vo trong cc cuvet ph hp 19 x 150 mm, thm lng nc va n 10 ml. Thm vo 4 ml dung dch natri metavanadat 5% va chun b v 1 ml acid acetic vo mi cuvet. (Ch : s dng cc dung dch ny trong 10 pht sau khi xut hin mu). Chun b mu trng theo cch tng t, s dng 10 ml nc thay th dung dch acid tartric. t trn thit b hp th bng 0 i vi mu trng, sau o hp th ca 4 dung dch acid tartric bc sng 520 nm vi my quang ph thch hp hoc my so mu quang in c trang b knh lc 520 nm. T cc kt qu thu c, v ng chun trn h to vung gc vi trc tung biu din hp th v trc honh biu din hm lng acid tartric (mg). Chun b mu th: Cn chnh xc 4 g mu cho vo bnh Erlenmeyer dung tch 250 ml v thm vo 80 ml dung dch KOH 0,5N v 0,5 ml phenolphthalein TS. Ni bnh vi mt sinh hn khng kh c chiu di t nht 65 cm v gia nhit hn hp trn tm bn nng trong 2,5 gi. Thm vo hn hp nng acid phosphoric 10% cho n khi c phn ng acid r rt pht hin c bng giy th congo. Li kt ni sinh hn kh v gia nhit cho n khi acid bo ha lng v trong. Lm ngui v chuyn hn hp vo mt phu chit dung tch 250 ml bng cc phn nh nc v chloroform. Chit tch cc acid bo c gii phng ra bng 3 ln lin tip 25 ml nc v cho phn nc ra vo phu chit cha lp nc. Chuyn cc thnh phn c trong phu chit th nht vo cc 250 ml, gia nhit trn chu hi nc loi cc vt chloroform, lc qua giy lc mn vo bnh nh mc dung tch 500 ml v cui cng pha ti th tch 500 ml bng nc (Dung dch I). Dng pipet ly 25 ml dung dch ny cho

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vo bnh nh mc dung tch 100 ml v pha ti th tch 100 ml bng nc (Dung dch II). Cch tin hnh: Chuyn 10 ml dung dch II c chun b trong phn Chun b mu th vo cuvet 19 x 150. Thm vo 4 ml dung dch natri metavanadat 5% va chun b v 1 ml acid acetic. (Ch : s dng cc dung dch ny trong 10 pht sau khi xut hin mu). Chun b mu trng theo cch tng t, s dng 10 ml nc thay th dung dch acid tartric. t trn thit b o hp th bng 0 i vi mu trng, sau o hp th ca 4 dung dch acid tartric bc sng 520 nm vi my quang ph thch hp hoc my so mu quang in c trang b knh lc 520 nm. T ng chun xc nh hm lng acid tartric (mg) trong dung dch pha cui cng, nhn hm lng acid tartric xc nh c vi 20 v chia cho trng lng mu ban u thu c % acid tartric. Cht khng Cch tin hnh ny xc nh nhng cht ha tan trong cht bo m x phng ha khng c x phng ha bi kim nhng c ha tan trong dung mi cht bo thng thng. Cch tin hnh: Cn chnh xc 5 g mu cho vo bnh dung tch 250 ml, thm dung dch c 2 g KOH trong 40 ml alcol v un si nh c sinh hn ngc trong 1 gi. Chuyn ton b hn hp cha trong bnh vo ng ong c np y thy tinh (chiu cao khong 30 cm, ng knh 3,5 cm v c vch nh mc 40, 80 v 130 ml). Ra bnh bng alcol ph hp n th tch 40 ml trong ng ong, thm nc m, sau bng nc lnh cho n khi tng th tch t 80 ml. Cui cng ra bnh bng vi ml ether du ha, cho phn dung mi ra vo ng ong, lm mt hn hp trong ng ong n nhit phng v thm 50 ml ether du ha. y np, lc ng ong mnh t nht trong 1 pht, hai lp tch ra trong. Ht lp bn trn hon ton nu c th m khng ht bt k mt cht dch no ca lp di, ly phn ether cho vo trong mt phu tch dung tch 500 ml. Lp li qu trnh trch ly v ht dch t nht 6 ln bng cc phn 50 ml ether du ha, lc mnh mi ln. Ra hn hp phn trch ly bng cc phn 25 ml alcol 10% v lc mnh cho n khi nc ra c trung Ha s dng cht ch th phenolphthalein, v loi b phn nc ra. Chuyn phn ether tch c vo cc bit khi lng v ra phu tch bng 10 ml ether, sau cho nc ra vo cc.

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C ether trong cch thy n kh v lm kh phn cn n khi lng khng i, tt nht 75 - 800C p sut chn khng khng qu 200 mm Hg, hoc 1000C p sut thng trong 30 pht. Lm mt trong bnh ht m v cn thu c khi lng cha hiu chnh ca cht khng x phng. Xc nh hm lng acid bo trong cn bng cch sau: ha tan cn trong 50 ml alcol m (cha phenolphthalein TS v trc c trung ha bng NaOH ti mu hng nht), chun bng dung dch NaOH 0,02N cho n khi xut hin mu hng nht. Mi ml dung dch NaOH 0,02N tng ng vi 5,659 mg acid bo, tnh theo acid oleic. Hiu s khi lng cha hiu chnh ca cht khng x phng v khi lng acid bo tnh c t php chun trn l khi lng cht khng x phng ha trong mu th. Ch s acid Cn chnh xc 1 g mu v ha tan trong 20 ml ethanol 95% nng, trc c trung ha c s dng 0,5 ml cht ch th phenolphthalein TS. Lm ngui dung dch sau trung ha bng cch chun vi dung dch KOH 0,01N trong ethanol. 0,561 x th tch dung dch KOH dng chun (ml) Ch s acid = --------------------------------------------------------------Khi lng mu (g) Gi li dung dch c trung ha cho phn nh lng. 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. Thm chnh xc 50 ml dung dch KOH 0,1N trong ethanol vo dung dch c trung ha trn t phn xc nh ch s acid v un si u trong 2 pht. Lm ngui dung dch v chun d KOH bng B ml dung dch HCl 0,1N. Thc hin xc nh vi mu trng ( bng A ml dung dch HCl 0,1N). 5,611 x (A - B) Ch s ester = ----------------------------------------Khi lng mu (g)

6.2. nh lng

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119

Ph lc 18 YU CU K THUT V PHNG PHP TH I VI SORBITAL MONOSTEARAT 1. Tn khc, ch s 2. nh ngha Ch s C.A.S Cng thc cu to INS 491 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid stearic thc phm. 1338-41-6 Gm acid palmitic este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Sorbitol 1,4-Sorbitan Isosorbid 3. Cm quan Ht hay mnh c mu kem nht n vng nu hoc dng sp rn, c mi nh c trng. 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan Ti nhit cao hn im chy, tan trong toluen, dioxan, carbon tetraclorid, ether, methanol, ethanolv anilin; khng tan trong ether du ha v aceton, khng tan trong nc lnh nhng c th phn tn trong nc m.Ti nhit trn 50oC tan dng sng m trong du khong v ethyl acetat. Khong nhit 50-52oC. ng c Hp th hng Ph hng ngoi ca mu th c trng cho este mt phn ca acid bo vi polyol ngoi 5.2. tinh khit Nc Khng c qu 1,5% (phng php Karl Fischer). Ch s acid Khng c thp hn 5 v khng c qu 10. Ch s x Khng c thp hn 147 v khng c qu 157. phng ha

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Ch s Khng c thp hn 235 v khng c qu 260. Hydroxyl Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng X phng ha 100g mu phi thu c khong 31,5g polyol v 73g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid. 6. Phng php th 6.1. tinh khit 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. 6.2. nh Tin hnh theo hng dn trong chuyn lun xc nh hm lng sorbitan ester (JECFA monograph 1 - Vol.4 ). lng

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Ph lc 19 YU CU K THUT V PHNG PHP TH I VI SORBITAN TRISTEARAT INS 492 ADI=0-25mg/kg th trng 2. nh ngha Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid stearic thc phm. Ch s C.A.S 26658-19-5 Cng thc cu Gm acid stearic este ha vi polyol dn xut t sorbitol bao gm to cc loi sau: 1. Tn khc, ch s

Sorbitol 1,4-Sorbitan Isosorbid 3. Cm quan Ht hay mnh c mu kem nht n vng nu hoc dng sp rn 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan Kh tan trong toluen, ether, carbon tetraclorid, ethyl acetat; c th phn tn trong ether du ha, du khong, du thc vt, aceton v dioxan; khng tan trong nc, methanol v ethanol (th theo hng dn ti JECFA monograph 1 - Vol.4 ). Khong nhit 47-50oC. ng c 5.2. tinh khit Nc Khng c qu 1,5% (phng php Karl Fischer). Tro sulfat Khng c qu 0,5%. Ch s acid Khng c qu 15. Ch s x Khng c thp hn 176 v khng c qu 188. phng ha Ch s Khng c thp hn 66 v khng c qu 80. Hydroxyl

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Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng X phng ha 100g mu phi thu c khng thp hn 14g v khng c qu 21g polyol v khng thp hn 85g v khng c qu 92g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid. 6. Phng php th 6.1. tinh khit 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. 6.2. nh Cn 25g mu th, chnh xc n mg, chuyn vo bnh cu y lng trn 500 ml, thm 250 ml cn v 7,5g kali hydroxyd v trn u. Lp bnh cu vi sinh hn thch hp, un hi lu hn hp trong t 1-2 gi sau chuyn hn hp sang cc 800 ml, trng sch bnh bng 100 ml nc ct v gp dch ra vo cc. Cho bay hi ht cn trn b cch thy, thnh thong thm nc thay th lng cn bay hi. Dng qu trnh ui cn khi khng cn mi cn bay ra. Hiu chnh th tch cui cng cho 250 ml bng nc nng. Trung ha dung dch x phng vi acid sulfuric long (1/2), thm 10% d v un nng, khuy u cho n khi acid bo tch lp. Chuyn lp acid bo sang phu chit 500 ml, ra 3 hoc 4 ln, mi ln vi 20 ml nc nng loi ht cc polyol, gp dch ra vi pha nc cha polyol thu c t qu trnh x phng ha. Chit pha nc ny 3 ln 20 ml ether du ho/ln, ly lp ether du ha gp vi phn acid bo, cho bay hi ti kh trong mt a xc nh b, ngui v cn. Trung ha dung dch cha polyol bng dung dch kali hydroxyd 1/10 ti pH = 7, s dng pH k thch hp. Cho bay hi dung dch ny ti gn cn, tch phn polyol khi cc mui bng cch chit vi ln vi cn nng. Cho bay hi dch chit cn trn b cch thy ti kh trong mt a xc nh b, ngui v cn. Trnh kh qu trong qu trnh un nng. Tin hnh nh lng mt mu khc (25g) theo hng dn chuyn lun xc nh hm lng ester sorbitan xc nh % ester sorbitan.

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Ph lc 20 YU CU K THUT V PHNG PHP TH I VI SORBITAN MONOLAURAT 1. Tn khc, ch s 2. nh ngha Ch s C.A.S Cng thc cu to INS 493 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid lauric thc phm. 1338-39-2 Gm acid lauric este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Sorbitol 1,4-Sorbitan Isosorbid 3. Cm quan Dng lng nht snh c mu h phch, dng ht hay mnh c mu kem nht n vng nu hoc dng sp rn, c mi nh c trng. 4. Chc nng Cht nh ha, cht n nh 5. Yu cu k thut 5.1. nh tnh tan C th phn tn c trong nc nng v nc lnh. 5.2. tinh khit Nc Khng c qu 2,0% (phng php Karl Fischer). Tro sulfat Khng c qu 0,5%. Ch s acid Khng c qu 7. Ch s x phng Khng c thp hn 155 v khng c qu 170. ha Ch s Hydroxyl Khng c thp hn 330 v khng c qu 358. Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng X phng ha 100g mu phi thu c khng thp hn 36g v khng c qu 49g polyol, khng thp hn 56g v khng c qu 68g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid.

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6. Phng php th 6.1. tinh khit 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. 6.2. nh lng Cn 25g mu th, chnh xc n mg, chuyn vo bnh cu y trn 500 ml, thm 250 ml cn v 7,5g kali hydroxyd v trn u. Lp bnh cu vi sinh hn thch hp, un hi lu hn hp trong t 1-2 gi sau chuyn hn hp sang cc 800 ml, trng sch bnh bng 100 ml nc ct v gp dch ra vo cc. Cho bay hi ht cn trn b cch thy, thnh thong thm nc thay th lng cn bay hi. Dng qu trnh ui cn khi khng cn mi cn bay ra. Hiu chnh th tch cui cng cho 250 ml bng nc nng. Trung ha dung dch x phng vi acid sulfuric long (1/2), thm 10% d v un nng, khuy u cho n khi acid bo tch lp. Chuyn lp acid bo sang phu chit 500 ml, ra 3 hoc 4 ln, mi ln vi 20 ml nc nng loi ht cc polyol, gp dch ra vi pha nc cha polyol thu c t qu trnh x phng ha. Chit pha nc ny 3 ln 20 ml ether du ha/ln, ly lp ether du ha gp vi phn acid bo, cho bay hi ti kh trong mt a xc nh b, ngui v cn. Trung ha dung dch cha polyol bng dung dch kali hydroxyd 1/10 ti pH = 7, s dng pH k thch hp. Cho bay hi dung dch ny ti gn cn, tch phn polyol khi cc mui bng cch chit vi ln vi cn nng. Cho bay hi dch chit cn trn b cch thy ti kh trong mt a xc nh b, ngui v cn. Trnh kh qu trong qu trnh un nng.

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Ph lc 21 YU CU K THUT V PHNG PHP TH I VI SORBITAN MONOOLEAT 1. Tn khc, ch s 2. nh ngha INS 494 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid oleic thc phm (R). Thnh phn ln nht l 1,4-Sorbitan monooleat, thnh phn t hn l isosorbid monooleat, sorbitan dioleat v sorbitan trioleat. 1338-43-8 Gm acid oleic este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Ch s C.A.S Cng thc cu to

Sorbitol 1,4-Sorbitan Isosorbid 3. Cm quan Dng st snh mu h phch, dng ht hay mnh c mu kem nht n vng nu hoc dng sp rn, c mi nh. 4. Chc nng Cht nh ha, cht n nh 5. Yu cu k thut 5.1. nh tnh tan Ti nhit cao hn im chy, tan trong ethanol, ether, ethylacetat, anilin, toluen, dioxan, ether du ha, carbon tetraclorid; khng tan trong nc lnh nhng c th phn tn trong nc m. Ch s iod Phn acid oleic thu c bng cch x phng ha sorbitan monooleat trong qu trnh nh lng c ch s iod t 80 n 100. 5.2. tinh khit Nc Khng c qu 2% (phng php Karl Fischer). Tro sulfat Khng c qu 0,5%. Ch s acid Khng c qu 8. Ch s x Khng c thp hn 145 v khng c qu 160. phng ha

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Ch s Khng c thp hn 193 v khng c qu 210. Hydroxyl Ch Khng c qu 2,0 mg/kg 5.3. Hm lng X phng ha 100g mu phi thu c khng thp hn 28g v khng c qu 32g polyol, khng thp hn 73g v khng c qu 77g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid. 6. Phng php th 6.1. tinh khit 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. 6.2. nh lng Cn 25g mu th, chnh xc n mg, chuyn vo bnh cu y trn 500 ml, thm 250 ml cn v 7,5g kali hydroxyd v trn u. Lp bnh cu vi sinh hn thch hp, un hi lu hn hp trong t 1-2 gi sau chuyn hn hp sang cc 800 ml, trng sch bnh bng 100 ml nc ct v gp dch ra vo cc. Cho bay hi ht cn trn b cch thy, thnh thong thm nc thay th lng cn bay hi. Dng qu trnh ui cn khi khng cn mi cn bay ra. Hiu chnh th tch cui cng cho 250 ml bng nc nng. Trung ha dung dch x phng vi acid sulfuric long (1/2), thm 10% d v un nng, khuy u cho n khi acid bo tch lp. Chuyn lp acid bo sang phu chit 500 ml, ra 3 hoc 4 ln, mi ln vi 20 ml nc nng loi ht cc polyol, gp dch ra vi pha nc cha polyol thu c t qu trnh x phng ha. Chit pha nc ny 3 ln 20 ml ether du ho/ln, ly lp ether du ho gp vi phn acid bo, cho bay hi ti kh trong mt a xc nh b, ngui v cn. Trung ha dung dch cha polyol bng dung dch kali hydroxyd 1/10 ti pH = 7, s dng pH k thch hp. Cho bay hi dung dch ny ti gn cn, tch phn polyol khi cc mui bng cch chit vi ln vi cn nng. Cho bay hi dch chit cn trn b cch thy ti kh trong mt a xc nh b, ngui v cn. Trnh kh qu trong qu trnh un nng. Tin hnh nh lng mt mu khc (25g) theo hng dn chuyn lun xc nh hm lng sorbitan ester xc nh % sorbitan ester.

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Ph lc 22 YU CU K THUT V PHNG PHP TH I VI SORBITAN MONOPALMITAT 1. Tn khc, INS 495 ch s 2. nh ngha Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid palmitic thc phm Ch s C.A.S 26266-57-9 Cng thc cu Gm acid palmitic ester ha vi polyol dn xut t sorbitol bao to gm cc loi sau:

Sorbitol

1,4-Sorbitan

Isosorbid

3. Cm quan Dng sp rn c mu kem nht n vng nu, c mi nh. 4. Chc nng Cht nh ha 5. Yu cu k thut 5.1. nh tnh tan Ti nhit cao hn im chy, tan trong methanol, ether, ethylacetat, anilin, toluen, dioxan, ether du ha, carbon tetraclorid; khng tan trong nc lnh nhng c th phn tn trong nc m. ng c 45 - 470 Hp th hng Phi c ph hng ngoi c trng ca ester ca acid bo vi ngoi polyol 5.2. tinh khit Nc Khng c qu 1,5% (phng php Karl Fischer). Ch s acid Khng c thp hn 4 v khng c qu 7,5. Khng c thp hn 140 v khng c qu 150. Ch s x phng ha Ch s Khng c thp hn 270 v khng c qu 305. Hydroxyl

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Ch 5.3. Hm lng

Khng c qu 2,0 mg/kg

X phng ha 100g mu phi thu c khong 37 g polyol v 65 g acid bo. Hm lng polyol phi vo khong 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid. 6. Phng php th 6.1. tinh khit 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. 6.2. nh Tin hnh theo hng dn trong chuyn lun xc nh hm lng sorbitan ester (JECFA monograph 1 - Vol.4). lng

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QCVN 4 - 23: 2011/BYT

QUY CHUN K THUT QUC GIA V PH GIA THC PHM - CHT TO BT National technical regulation on Food Additive - Foaming agents

Li ni u QCVN 4-23: 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.

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QUY CHUN K THUT QUC GIA V PH GIA THC PHM - CHT TO BT National technical regulation on Food Additive - Foaming agent 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 to bt 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 to bt 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 to bt: l ph gia thc phm c cho vo thc phm nhm to ra hoc duy tr s phn tn ng nht ca pha kh trong thc phm dng lng hoc dng rn. 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. II. YU CU K THUT, PHNG PHP TH V LY MU 1. Yu cu k thut v phng php th i vi cht chit xut t Quillaia s dng lm cht to bt c quy nh ti ph lc ban hnh km theo Quy chun ny. 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 ph lc.

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Cc phng php th c hng dn trong Quy chun ny khng bt buc phi p dng, c th s dng cc phng php th khc tng ng. 3. Ly mu theo hng dn ti Thng t s 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. Cht to bt 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 to bt Vic kim tra cht lng, v sinh an ton i vi cht to bt 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 cht to bt sau khi hon tt ng k bn cng b hp quy v bo m cht lng, v sinh an ton, ghi nhn ph hp vi cc quy nh ca php lut. V. T CHC THC HIN 1. Giao Cc An ton v sinh thc phm ch tr, phi hp vi cc c quan chc nng c lin quan hng dn trin khai v t chc vic thc hin Quy chun ny. 2. Cn c vo yu cu qun l, Cc An ton v sinh thc phm c trch nhim kin ngh B Y t sa i, b sung Quy chun ny. 3. Trng hp hng dn ca quc t v phng php th v cc quy nh ca php lut vin dn trong Quy chun ny c sa i, b sung hoc thay th th p dng theo vn bn mi.

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Ph lc YU CU K THUT V PHNG PHP TH I VI CHT CHIT XUT T QUILLAIA (DNG 1) Quillaja extract, Soapbark extract, Quillay bark extract, Bois de Panama, Panama bark extract, Quillai extract. ADI: 0 - 1mg/kg th trng (tnh cho c dng 1 v 2) INS 999i 2. nh ngha Cht chit xut t quillaia (dng 1) thu c t qu trnh chit vi nc v hoc g ca thn v cnh Quillaja saponaria Molina (h Rosaceae). Ch phm cha saponin triterpenoid trong ch yu l glycosid ca acid quillaic. Polyphenol v tanin l nhng thnh phn chnh, ngoi ra cn c mt vi loi ng v calci oxalat. Ch phm chit xut t quillaia (dng 1) thng mi dng lng hoc bt sy phun c cht mang nh lactose, maltitol hoc maltodextrin. Sn phm dng lng thng c bo qun bng natri benzoat hoc ethanol. Tn ha hc Saponin triterpenoid (quillaia saponin), glycosid ca acid quillaic M s C.A.S. 68990-67-0 Cc saponin dng monomer c khi lng phn t khong 1.800 Khi lng phn t 2.300, ph hp vi mt triterpen gm 8 - 10 monosaccharid. 3. Cm quan Dng lng mu nu hoc dng bt mu nu sng c nh hng 4. Chc nng Cht nh ha, cht to bt 5. Yu cu k thut 5.1. nh tnh tan Tan tt trong nc; khng tan trong ethanol, aceton, methanol v butanol. To bt Phi c phn ng to bt c trng. Sc k Thi gian lu ca pic chnh ca mu tng ng vi pic saponin chnh (QS-18) ca chun (m t trong phn Phng php th) Mu sc v Phi c mu sc v c c trng. c 5.2. tinh khit Nc Dng bt: Khng c qu 6% (theo phng php Karl Fischer) Gim khi lng Dng lng: 50 - 80% (sy 2 g mu nhit 1050C trong 5 gi) khi sy kh pH 3,7 - 5,5 (i vi nng dung dch 4%) 1. Tn khc, ch s

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Khng c qu 14% theo ch phm kh (dng 1 g mu i vi dng bt; i vi mu dng lng s dng phn cn li sau khi sy kh) Tanin Khng c qu 8% theo ch phm kh (m t trong phn Phng php th) Ch Khng c qu 2,0 mg/kg. 5.3. Hm lng Hm lng saponin: khng c thp hn 20% v khng c qu 26% tnh theo ch phm kh. 6. Phng php th 6.1. nh tnh To bt Ha tan 0,5 g mu dng bt trong 9,5 g nc hoc 1 ml mu dng lng trong 9 ml nc. Cho 1 ml hn hp v 350 ml nc vo trong ng ong loi 1.000 ml. Bt kn ng ong v lc mnh 30 ln, yn. Ghi li mc bt (ml) sau 30 pht. Bnh thng mc bt t khong 150 ml bt. Mu sc v i vi dng bt: ha tan 0,5 g mu trong 9,5 g nc. Dung dch khng c. hp th ca dung dch so vi nc bc sng c 520 nm phi nh hn 1,2. 6.2. tinh khit Cn 3,0 g mu dng bt hoc lng mu dng lng tng ng Tanin tnh theo lng ch phm kh sau khi sy. Ha tan mu trong 250 ml nc. iu chnh pH = 3,5 bng acid acetic. Sy 25 ml dung dch thu c nhit 1050C trong thi gian 5 gi v xc nh khi lng cht kh theo n v g (Wi). Trn 50 ml dung dch trn vi 360 mg polyvinyl polypyrrolidon, sau khuy trong 30 pht nhit phng; ly tm vi tc 8.000 vng/pht. Ly phn dung dch trong pha trn, sy nhit 1050C, thi gian 5 gi. Cn phn cht kh thu c (Wf, n v tnh g). Hm lng tanin (%) trong mu c tnh nh sau: % Tanin (theo ch phm kh) = 100 x (Wi - Wf/2)/Wi 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 Nguyn tc: Saponin QS-7,QS-17, QS-18 v QS-21 c tch bng HPLC pha o, kt qu nh lng c xem l tng hm lng saponin c trong cht chit xut t Quillaia (dng 1)

Tro

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Chun b mu: i vi mu dng bt: Cn 0,5 g mu v ha tan trong 9,5 g nc. Lc qua b lc c 0,2 m. Dch chit nc (khong 550 mg cht kh/ml): Cn 1 g mu v pha vi 9 g nc. Lc qua b lc c 0,2 m. Trong c hai trng hp trn, th tch mu khong 10 ml. Chun b mu chun: Cn 1,5 g saponin tinh sch (ca cc hng SuperSap, Natural Response, Chil; Quil-A, Superfos, an Mch hoc tng ng, bit hm lng saponin), ha tan trong 100 ml nc. Lc qua b lc c 0,2 m. iu kin HPLC: - Ct: Vydac 214TP54 (di 4,6 x 250 mm, l 5 m) hoc loi tng ng - Nhit ct: nhit phng - Bm: t ch gradient - Dung mi A: 0,15% TFA (acid trifloroacetic) trong nc dng cho HPLC - Dung mi B: 0,15% TFA (acid trifloroacetic) trong acetonitril dng cho HPLC - Gradient: Thi gian (pht) % Dung mi A % Dung mi B 0 70 30 40 55 45 45 70 30 - Tc dng: 1 ml/pht - Bc sng pht hin: 220 nm - Th tch bm mu: 20 l Tnh kt qu: Nng saponin (mg/ml) trong dung dch c chun b nh trn l: Cmu = (Amu/Achun)Cchun Trong : - Cmu: Nng saponin (mg/ml) trong mu th - Cchun: Nng saponin chun (mg/ml) c bm vo (v d: Cchun = 13,5 mg/ml nu hm lng saponin ca 1,5 g mu chun l 90%) - Amu v Achun: Tng din tch cc pic tng ng vi 4 loi saponin chnh (QS-7,QS-17, QS-18, QS-21) c trong mu cn

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phn tch v mu chun. (Tanin v polyphenol c tch gii trc saponin. Cc pic ca saponin xut hin sau pic chnh ca polyphenol - xem hnh ph lc). % Saponin c trong mu th c tnh nh sau: % Saponin = 100 x Cmu/(0,1 Wmu) Trong : - Wmu: Khi lng mu (mg) c ly chun b mu v 0,1 l nghch o ca th tch mu (10 ml). Sc k ca cht chun (15 mg cht kh/ml tng ng vi 13,5 mg saponin/ml)

Sc k ca cht chit xut t Quillaia (dng 1) (55 mg cht kh/ml)

- Ht -