I IHC HCQUC QUCGIA GIAH HNI NI BèA TRNG TRNGI IHC HCKHOA KHOAHC HCT TNHIấN NHIấN - V AnhTin Tun Nguyn Lung NH GI C CAFISH SODIUM S DNG KTNH THUT BENZOATE, KIM TRA PROPYL GALLATE, TARTRAZINE, AMARANTH, S HI NHP CA GEN IL6 PHN LP T NGI MONOSODIUM GLUTAMATE V FORMALDEHYDE TRONG BO GC PHễI GC CHUYN GEN TRấN Mễ T HèNH PHT TRIN PHễI NGA VN LUN VN THC S KHOA HC LUN VN THC S KHOA HC H Ni 2015 H Ni 2013 I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - V Anh Tun NH GI C TNH CA SODIUM BENZOATE, PROPYL GALLATE, TARTRAZINE, AMARANTH, MONOSODIUM GLUTAMATE V FORMALDEHYDE TRấN Mễ HèNH PHT TRIN PHễI C NGA VN Chuyờn ngnh: Sinh hc Thc nghim Mó s: 60.42.0114 LUN VN THC S KHOA HC NGI HNG DN KHOA HC: PGS TS Nguyn Lai Thnh TS Hong Th M Hnh H Ni 2015 LI CM N u tiờn, tụi xin gi li cm n chõn thnh nht n PGS TS Nguyn Lai Thnh, ngi thy ó thu nhn, hng dn, truyn t cho tụi nhng kin thc v kinh nghim sut thi gian hc nghiờn cu ti phũng thớ nghim Thy ó luụn theo sỏt, ch bo cho tụi nhng gúp ý quý bỏu tụi cú th hon thnh tt cụng vic Tụi cng rt bit n TS Hong Th M Hnh ó ch bo, hng dn tụi kin thc, k nng mi tụi hon thin c lun tt nghip ny Tụi xin gi li cm n n NCS inh Duy Thnh, ngi ó ch dy, hng dn cho tụi t nhng ngy u tụi vo lm vic ti phũng, truyn t cho tụi nhng kinh nghim cụng vic cng nh cuc sng Tụi xin by t lũng bit n sõu sc n cỏc thy cụ cụng tỏc ti b mụn Sinh hc T bo cng nh cỏc thy cụ khoa Sinh hc ó truyn t cho tụi nhng kin thc c s, lm nn tng cho vic thc hin nghiờn cu Tụi xin cm n tt c cỏc anh ch, cỏc bn v cỏc em sinh viờn ang hc v cụng tỏc ti phũng Cụng ngh T bo ng vt - Trung tõm Nghiờn cu Khoa hc S sng, c bit l hc viờn cao hc Lu Hn Ly ó luụn bờn cnh giỳp , h tr tụi thi gian thc hin nghiờn cu Tụi xin c chõn thnh cm n gia ỡnh, bn bố v ngi thõn, ó luụn bờn, ng viờn cho tụi cú thờm ngh lc vt qua c khú khn sut thi gian qua H Ni, thỏng 12 nm 2015 Hc viờn V Anh Tun MC LC DANH MC HèNH DANH MC BNG BNG Kí HIU V CH VIT TT M U CHNG TNG QUAN 1.1 THC TRNG S DNG HểA CHT 1.2 PH GIA THC PHM 1.2.1 S lc v ph gia thc phm 1.2.2 Cỏc loi ph gia thc phm c s dng nghiờn cu 1.2.2.1 Cht bo qun - Sodium benzoate 1.2.2.2 Cht chng ụ xy húa - Propyl gallate 1.2.2.3 Cht to mu vng - Tartrazine 1.2.2.4 Cht to mu - Amaranth 1.2.2.5 Cht iu v - Monosodium glutamate 1.2.2.6 Cht bo qun ó b cm s dng - Formaldehyde 10 1.3 CC Mễ HèNH NH GI C TNH HểA CHT 10 1.3.1 S dng ng vt thớ nghim ỏnh giỏ c tớnh húa cht 11 1.3.2 Mụ hỡnh thay th ng vt thớ nghim ỏnh giỏ c tớnh húa cht 12 1.3.2.1 Mụ hỡnh ỏnh giỏ c tớnh s dng t bo nuụi cy in vitro 12 1.3.2.2 Mụ hỡnh phụi cỏ nga ỏnh giỏ c tớnh 12 CHNG PHNG PHP NGHIấN CU .18 2.1 I TNG NGHIấN CU 18 2.2 DNG C, HểA CHT, THIT B 18 2.2.1 Dng c, thit b 18 2.2.2 Húa cht 19 2.3 QUY TRèNH TH NGHIM 21 2.3.1 Quy trỡnh nuụi cỏ b m v thu phụi 21 2.3.2 Phi nhim vi húa cht 21 2.3.3 ỏnh giỏ s nh hng ca cỏc cht ph gia ti s phỏt trin phụi da trờn hỡnh thỏi v sc sng 23 2.3.4 ỏnh giỏ s nh hng ca cỏc cht n nhp tim phụi/u th cỏ nga 25 2.3.5 CHNG Phõn tớch thng kờ 25 KT QU V THO LUN .27 3.1 S BIN I HèNH THI V SC SNG CA PHễI C NGA VN KHI PHI NHIM VI CC CHT PH GIA 29 3.1.1 Hỡnh thỏi phụi cỏ nga i chng 29 3.1.2 Hỡnh thỏi phụi phi nhim vi nhúm cht to mu 30 3.1.2.1 Phi nhim vi cht to mu vng Tartrazine (E102) 30 3.1.2.2 Phi nhim vi cht to mu Amaranth (E123) .34 3.1.3 Hỡnh thỏi phụi phi nhim vi nhúm cht bo qun 36 3.1.3.1 Phi nhim vi sodium benzoate (E211) .36 3.1.3.2 Phi nhim vi propyl gallate (E310) 39 3.1.4 Nhúm cht iu v - Monosodium glutamate (E621) 41 3.1.5 Cht bo qun ó b cm s dng Formaldehyde (E240) 43 3.1.6 S nh hng ca húa cht n t l phụi n 47 3.2 S NH HNG CA CC HểA CHT TH NGHIM N NHP TIM PHễI C NGA VN 49 3.3 C TNH CA CC CHT PH GIA TRONG NGHIấN CU 51 KT LUN V KIN NGH 57 TI LIU THAM KHO 59 DANH MC HèNH Hỡnh 1.1 S tng ng v gen hot ng gia ngi, chut, g v cỏ nga 13 Hỡnh 1.2: So sỏnh kt qu t th nghim c tớnh trờn cỏc loi cỏ v th nghim trờn phụi cỏ nga .15 Hỡnh 1.3 Cỏc lnh vc nghiờn cu s dng phụi cỏ nga v s lng nghiờn cu c cụng b t 1992 n 2015 .17 Hỡnh 2.1 Hỡnh thỏi cỏ nga trng thnh 18 Hỡnh 2.2 Phõn b nng trờn a 24 ging 22 Hỡnh 2.3 Hỡnh thỏi phụi cỏ nga mt s giai on theo Kimmel 24 Hỡnh 3.1 Hỡnh thỏi phụi v u th cỏ nga i chng .29 Hỡnh 3.2 Phụi cỏ nga phi nhim vi Tartrazine nng g/l 30 Hỡnh 3.3 T l phụi cũn sng v t l phụi d dng phi nhim vi Tartrazine thi im 24, 48 v 72 gi sau th tinh 31 Hỡnh 3.4 Phụi cỏ nga phi nhim vi Tartrazine 32 Hỡnh 3.5 T l phụi cũn sng v t l phụi d dng phi nhim vi Tartrazine thi im 96 gi sau th tinh 34 Hỡnh 3.6 T l phụi cũn sng v t l phụi d dng phi nhim vi Amaranth 35 Hỡnh 3.7 Phụi cỏ nga sau 96 gi phi nhim vi Amaranth 36 Hỡnh 3.8 Phụi cỏ nga phi nhim vi Sodium benzoate 37 Hỡnh 3.9 T l phụi sng v t l phụi d dng phi nhim vi Sodium benzoate 38 Hỡnh 3.10 T l phụi sng v t l phụi d dng phi nhim vi Propyl gallate 39 Hỡnh 3.11 Phụi cỏ nga phi nhim vi Propyl gallate 40 Hỡnh 3.12 Phụi cỏ nga phi nhim vi Monosodium glutamate 41 Hỡnh 3.13 T l phụi cũn sng v t l phụi d dng phi nhim vi Monosodium glutamate 42 Hỡnh 3.14 Phụi cỏ nga phi nhim vi Formaldehyde 44 Hỡnh 3.15 T l phụi sng v t l phụi d dng phi nhim vi Formaldehyde 45 Hỡnh 3.16 T l cỏc loi d dng quan sỏt c 96 gi sau th tinh .46 Hỡnh 3.17 T l phụi n thi im 72 gi sau th tinh 48 Hỡnh 3.18 Nhp tim/phỳt ca u th cỏ nga phi nhim vi cỏc ph gia 50 Hỡnh 3.19 Ch s LC50 thu c cỏc thi im ca cỏc cht .53 Hỡnh 3.20 Cỏc ch s c hc ca cỏc cht thi im 96 gi sau th tinh 54 Hỡnh 3.21 Ch s c hc TI ca cỏc cht thi im 96 gi sau th tinh .55 DANH MC BNG Bng 1.1 Phõn loi ph gia thc phm Bng 2.1 Dng c, thit b c s dng lun 18 Bng 2.2 Cỏc loi húa cht c th nghim 20 Bng 2.3 Mt s tiờu ỏnh giỏ s phỏt trin phụi cỏ nga 23 Bng 3.1 Cỏc di nng thớ nghim ca cỏc cht 28 Bng 3.2 Liu lng an ton s dng hng ngy chp nhn c .56 BNG Kí HIU V CH VIT TT T vit tt Vit y ADI Acceptable Daily Intake - Lng n vo hng ngy chp nhn EC50 c Median effective concentration - Nng gõy nh hng 50% cỏ LC50 th thớ nghim Median lethal concentration - Nng gõy cht 50% cỏ th thớ LOEC nghim Lowest observed effect concentration - Nng thp nht quan sỏt NOEC thy nh hng ỏng k so vi i 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