TCVN TIEU CHUAN QUOC GIA TCVN 8138 : 2009 ISO 5553 : 1980 Xuat ban lan 1 THIT VA SAN PHAM THIT - PHAT HIEN POLYPHOSPHAT Meat and meat products ~ Detection of polyphosphates HA NOI 2009ICVN 8138 : 2009 Lei néi dau TCVN 8138 : 2009 hon toan twong duong voi ISO 5553 : 1980; TCVN 8138 : 2009 do Ban kf thuat titu chudn quéc gia TCVN/TC/FS- Thit va sn phdm thit bien soan, Téng cyc Tiéu chudn Do ludng Chat lugng 48 nghi, Bd Khoa hoc va Céng nghé céng bé.TIEU CHUAN QUOC GIA TCVN 8138 : 2009 Thit va san pham thit - Phat hién polyphosphat Meat and meat products — Detection of polyphosphates 1 Pham vi ép dung Tiéu chudn nay qui dinh phyong phap phat hién phosphat d4m dac trong thit va san pham thit bang sdc. ky lop mong. 2. Linh vye 4p dung Vi cdc polyphosphat dugc thuy phan ty tir bang enzym co mat trong thit hoc san phdm thit va trong qua trinh xi ly nhigt thit hodc san phdm thit, nén tiéu chudn nay chi 4p dung dé phat hién cac polyphosphat bé.sung van con cé mat trong mau tai thoi diém kiém tra. 3. Tailigu vign din Cac tai ligu vién dan sau rat can thiét cho viéc 4p dung tiu chudn nay. B6i véi cac tai ligu vién dan ghi nam céng bé thi 4p dyng phién ban dugc néu. Bi voi cac tai liéu vign d&n khong ghi nam céng bé thi 4p dung phién ban méi nhét, bao gdm cd cac siva di, bd sung (néu cb) 180 3100, Thit va sdn phdm thit - Lay mau ”. 4 Nguyén tac Chiét thit hode san phdm thit bang axit tricloaxetic. Lam trong dich chiét thu dug nay bang hén hop. etanol/dietyl ete. Tach cdc phosphat bing sic ky lép méng va phat hién polyphosphat bang cach phun thude thir 8 hién mau, 180 3100 68 durge thay th& bang hai tiéu chun 1SO 3100-1 : 1991 (68 dude bien soan thanh TCVN 4833-1 » 2002 ) va 180 3100-2 : 1991 (€a duoc bién soan thanh TCVN 4833-2 * 2002) Hin nay, ISO 3100-1 - 1891 68 bj hdy va éuge thay tné bang ISO 17604 : 2003 (éurgc bién soan thénh TCVN 7925: 2008 Vi sink véttrong thre pdm va thi an chin nubs - Phizong php ty mu than tit toi dé phan tich w sinh va)5 Thuéc thir TAt c& cae thude thir dugc st dung phai [a loai phan tich. Nu@e dugc sir dung phai la nude cat hoae it nat la nude c6 46 tinh khiét trong durong. CANH BAO - Tuan thi tét ca cae cha ¥ vé an toan thich hyp khi tién hanh theo cdc quy trinh quy dinh trong tiéu chuan nay. 5.1 Axittricloaxetic 5.2 Ete diety! 5.3. Etanol, 95 % (thé tich) 8.4 Bot xenluloza, ding cho sac ky lop méng. 5.8 Tinh bot co thé hoa tan 5.6 Hén hyp déi chieng Hoa tan trong 100 mi nue: — 200 mg natri dihydro phosphat ngam mét phan tir nude (NaH;PO,H;0), — 300 mg tetranatri diphosphat ngam mudi phan tl nuéc (NaxP;07.10H,0); — 200 mg pentanatri triphosphat (NasP3O.0) va — 200 mg natri hexametaphosphat (NaPOs), [x>10] Hén hop déi ching khi durge bao quan & 4 °C co thé bén durgc it nhdt 4 tuan. 5.7 Dung méitrién khai Trén 140 ml isopropyl alcohol, 40 ml dung dich axittricloaxetic 135 gi, 0.6 ml amoni hydroxit a9 = 0,80 g/ml dung dich khoang 25 % (khéi lueng). Git? dung méi trong chai kin khi. §.8 Thudéc thir dang phun song! Tron du cdc thé tich bang nhau cla dung dich amoni molybdat ngam bén phan tu nude [(NH.)¢M0;0,4.4H,0)] 75 gil va axit nitric d&m G3¢ (om = 1,40 giml) va hod tan 10 g axit tartaric trong 100 mi hén hop nay. ‘Chudn bj thude thr trong ngay sir dung.5.9 Thudc thir dang phun svong II Hoa tan 0,5 g axit 1-amino-2naphtol-Asulphonic trong 195 mi hdn hep cia nari disulphit (natri metabisulphit; NazSz0s) 150 gf va 5 mi dung dich natri sulphit (Na,SO3) 200 g/l. Hoa tan 40 g natri axétat ngam ba phan ti: nude (NaOOCCH;.3H,0) trong hén hop nay. Bao quan thuéc thir nay trong chai mau nau day kin 8 trong td lanh. Logi bé dung dich sau 1 tuan. 6 Thiét bj, dung cy Sir dung cac thiét bi, dung cy cla phéng tht nghiém théng thudng, trir khi co qui dinh khdc va cy thé nhy sau: 6.1. T4m thuy tinh, 48 khir chat béo, 10 cm x 20 cm. 6.2 Dyng cy dan mau, ding 48 chudn bj cdc Iép c6 bé day 0,25 mm. Néu khéng sn cé dyng cy nay thi co thé ding cac tém lop méng ban san, c6 46 day Iép 0,25 mm voi didu kién a tinh bdt dugc sur dung lam cht két dinh. Cac tém chia thach cao (canxi sulfat) la khéng phu hgp. 6.3. May tron phéng th nghigm. 6.4 Tu hut dm. 6.5 May xay thjt bang co, c& phong thir nghiém, 6 gan tam duc I, dudng kinh 5 khong qua 4 mm. 6.6 Gidy Ic g4p nép, dong kinh 15 cm. 6.7 Micro-pipet, 1 :1, hoc micro-xyranh cé vit dinh lvgng va dau thuy tinh uén cong. 6.8 Bé thuy tinh dugc lot gidy, kich thud thich hgp, cd np day kin khi, dung cho sac ky lop méng, 6.9 May sdy téc, co kha nng cung cAp Iudng khéng khi & nhiét 49 phdng hode dong khi 4m. 6.10 Binh phun. 6.11 Ti sdy, cO kha nang kiém soat durge 6 60°C. 7 Lay mau 7.4 Léy it mhdt 200 g mAu phong thir nghiém. Xem ISO 3100. 7.2. Chuan bj mau thir trong ngay nhan mau phéng thir nghiém,8 Cach tién hanh 8.1 Chudn bj cac tam lop mang Hoa tan 0,39 tinh bét (5.5) trong 90 mi nuéc séi. 8 ngusi, thém khong 15 g bét xenluloza (5.4) va dng hod trong may trn phéng thir nghiém (6.3) trong 1 min Dan hén hep nay lén t4m thuy tinh (6.1) bang dyng oy dan (6.2), diéu chinh dé thu duge lp day 0,25 mm SAy khd cdc tém Ip méng dugc 62 yén trong 60 min bang khdng Khi & nhiét d6 phdng va cudi cung bang khéng khi néng trong 10 min & 100 °C Bao quan céc tam nay trong tu hut Am (6.4). Cach khac, c6 thé si dung cdc tm lp méng s&n c6 (6.2). 8.2. Chuan bj mau thir ‘Bong hoa mau bang cach xay it nhAt hai lin bang may xay (6.5) va trén déu. Git dy mu trong chai kin khi, day nap chai va bao quan trong td anh, néu cn. Phan tich m&u cang sém cang t6t, chi trong vong §h. 8.3 Chuan bj huyét thanh 8.3.1 Lam vot 50 g mau thi? (8.2) bang 15 mi nude & 40 °C dén 60 °C trong céc co mé, ding dao tron ho&c dua khudy dau det 68 rn cho dén khi thu duc khéi lung déng nhat, trong khdng qua § min. 8.3.2 Thém 10g axit tricloaxetic (5.1) va tron kg lai. 8.3.3 Cho ngay vao td Janh trong 1 h va gan qua gidy loc gp nép (6.6) 44 thu ldy huyét thanh 44 tach. 8.3.4 Néu dich Ioc bi dyc, thi lac mot lan voi mét thé tich tuong ty clla ete dietyl (5.2). Ding mét pipet 1nhé leai bd Iep ete va thm mét thé tich tuong tyr eva etanol (5.3) vao pha léng. Lac 1 min, Bé yén hén hop trong vai phut va loc qua gidy loc gap nép (6.6). 8.4 Tach bang sic ky 8.4.1 Rét dung mdi tridn khai (5.7) véo trong bé trién khai (6.8) voi d¢ sau tir S mm aén 10 mm va day nap. Dé yén trong it nhat 30 min & nhiét 46 mdi trong, tranh anh nang mat trai va gid lua. 8.4.2 Chdm 3 ul huyét thanh, hoge 6 jl néu thye hién quy trinh lam trong 8.3.4, vao lop xenluloza (8 1) trén dudng ké bat chi cach day khoang 2 cm, Gif céc éiém nhd bang cach cham mét ian 1 ydDung may sdy t6c (6.9) & ché 66 dm dé sdy khd. CHU THIGH. Phai trénh khi ndng vi nguy co am thu phn phosphat 8.4.3 Tuong ty, chim Sul hén hop déi ching (6.6) Ién tm lop méng, cdch diém mau tw 1 cm dén 1.5 cm, nhung cé cling khong cach chinh xéc tinh tir Gay. 8.4.4 Md nap bé va cdn than dat ngay tdm xenluloza vao bé. Day ngay nap. Trién khai tm lop mong @ nhigt 49 méi trvéng, trdnh anh ndng mat trai va gid Ia. 8.4.5 Tiép tuc trién khai cho é&n khi mat dung mdi dang Ién khong 10 cm ké tiy €ueng but chi. Lay tdm lép méng ra khéi bé va s4y 10 min trong tu sdy (6.11) & 60 °C, hoe 30 min & nhiét 46 méi trong, hoac trong ludng khi lanh. 8.5 Phathign phosphat 8.5.1. Dat tam lop méng theo phuong thang ding dui ti hut, phun nhe va dau Ién tam lop mong thudc th dang phun surong | (5.8). Néu cé polyphosphat, thi xudt hién ngay cae chém vang 8.5.2 Dung may say toc (6.9) & ché d6 dm 8 sdy khé tam lop mong, Sau do s4y khé trong tu sdy it hat 1h & 100 °C dén khi loai bé han axit nitric. Lay tm lop méng ra khdi WW say va kiém tra dé chdc chan khdng cén mui hang cla axit nitric. 8.6.3 Bé tm lop méng ngugi dén nhigt 4 phong va dat dudi td hut. Phun nhe va déu len tm lop méng thue thir dang phun song Il (5.9) Néu ¢6 polyphosphat, thi xuat hién ngay cae chém mau xanh nuéc bién. Tuy ntitn, evdng dd mau aia cdc chdm xanh dupe 190 1a boi CHU THICH khong nhét tniét phat phun bang thube thi thude thir nay 68 gidp cho vige phat hién durge a8 dang, 9 Dién gia So sénh khodng c&ch di chuyén cua cdc 6iém phosphat tir m&u so véi cdc diém phosphat khac cla hon hop déi cheng. Luén Iuén cd mat mot diém ch4m ortophosphat, néu mau ¢b chia cdc phosphat dam dic, thi sé théy duge diém diphosphat va/ hoa cac diém phosphat triing hyp cao hon. Gia tri Ry cla cdc phosphat trong hdn hop déi ching 1a: ‘orthophosphat tU 0,80 dén 0,90TCVN 8138 : 2009 diphosphat (pyrophosphat) tu 0,50 dén 0,60 triphosphat 170,25 dn 0,35 hexametapolyphosphat (mudi Graham's) 0,0 Nhin chung, cac gia tri Rr cUa cdc polyphosphat trong chat chiét thit va sén phdm thit hoi thdp hon. CHU THICH Vide higu chink chénh Iéch ede gid tr Rr cia phasphat trong chat chiét mu va trong hn hop Abi ching C5 thé thu duge bing céch cho cht chiét mu thit tuoi lan cing mét thm lop méng, Vi thit toi chi chiza céc monophosphst, nén hin tram higu chinh o6 thé thu durge bing céch so sénh Khoding di chuyén ca didm chun ny v6i diém twong ing ota hén hop obi chong, 10 Bao céo thir nghigm Bao céo thir nghigm cn chi rS phurong phap ¢& sir dung va két qué thu durge. Bao cao thir nghiém ciing cin dé cap dén moi chi tiét thao tac khéng dugc quy dinh trong tiéu chudn nay, hoe dec xem [a ;, cling vei mgi tinh hudng bat thuéng cé thé anh hudng dén két qua. tuy Bao cao thir nghiém cing phai bao gdm moi chi tit cAn thiét 48 nhan biét day du vé mau. 10