Nghiên cứu phân tích một số amin thơm trong các sản phẩm da bằng phương pháp LC MS MS

I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - T TH NGC NH NGHIấN CU PHN TCH MT S AMIN THM TRONG CC SN PHM DA BNG PHNG PHP LC/MS/MS LUN VN THC S KHOA HC H Ni Nm 2016 I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - T TH NGC NH NGHIấN CU PHN TCH MT S AMIN THM TRONG CC SN PHM DA BNG PHNG PHP LC/MS/MS CHUYấN NGNH: HểA PHN TCH M S: 60440118 LUN VN THC S KHOA HC Ngi hng dn khoa hc: PGS.TS NGUYN VN RI H Ni Nm 2016 LI CM N Vi lũng bit n sõu sc, tụi xin chõn thnh cm n PGS.TS Nguyn Vn Ri ó tn tỡnh hng dn, giỳp v to iu kin cho tụi hon thnh Lun ny Tụi xin gi li cm n chõn thnh ti cỏc anh ch ng nghip cụng tỏc ti Trung tõm Phõn tớch v Cụng ngh Mụi trng Vin Nghiờn cu Da giy ó to iu kin tt nht cho tụi c hc v nghiờn cu Tụi cng xin gi li cm n ti cỏc thy cụ ging dy ti Khoa Húa hc Trng i hc Khoa hc T nhiờn i hc Quc gia H Ni cựng th lp cao hc K24 ó giỳp ng viờn tụi sut quỏ trỡnh hc v nghiờn cu ti trng Cui cựng tụi xin gi li cm n n gia ỡnh, bn bố nhng ngi ó luụn ng viờn, khuyn khớch v c v tụi sut quỏ trỡnh hc v nghiờn cu H Ni, thỏng 12 nm 2016 Hc viờn T Th Ngc nh MC LC M U CHNG 1: TNG QUAN 1.1 Tng quan v thuc nhum azo 1.1.1 Gii thiu v thuc nhum azo 1.1.2 S chuyn hoỏ thuc nhum azo thnh cỏc amin thm 1.1.3 nh hng ca cỏc amin thm c gii phúng t thuc nhum azo n sc kho ngi 1.2 Tng quan hin trng s dng thuc nhum azo ngnh thuc da 1.2.1 Hin trng s dng thuc nhum azo trờn th gii 1.2.2 Hin trng s dng thuc nhum azo ti Vit Nam 1.3 Tng quan phng phỏp phõn tớch cỏc amin thm gii phúng t thuc nhum azo 11 1.3.1 Phng phỏp sc ký lng ph 11 1.3.2 Phng phỏp phõn tớch cỏc amin thm trờn th gii v Vit Nam 13 CHNG 2: THC NGHIM 24 2.1 Húa cht Thit b 24 2.1.1 Hoỏ cht chun 24 2.1.2 Dung mụi v hoỏ cht ph tr 24 2.1.3 Thit b phõn tớch 24 2.1.4 Thit b ph tr 25 2.2 Phng phỏp nghiờn cu 26 2.2.1 Phng phỏp k tha 26 2.2.2 Phng phỏp thc nghim 26 2.2.2.1 Kho sỏt iu kin ti u cho thit b sc ký lng ph 26 2.2.2.2 Xõy dng quy trỡnh x lý mu 27 2.2.2.3 Xỏc nhn giỏ tr s dng ca phng phỏp phõn tớch 28 2.2.2.4 So sỏnh phng phỏp phõn tớch mi xõy dng vi phng phỏp tiờu chun 32 CHNG 3: KT QU V THO LUN 34 3.1 Kho sỏt iu kin ti u cho thit b sc ký lng ph 34 3.1.1 Kho sỏt quỏ trỡnh tỏch sc ký 34 3.1.2 Kho sỏt cỏc thụng s ti u cho u dũ ph 38 3.1.3 Kho sỏt thi gian lu ca cỏc cht amin thm 42 3.2 Kho sỏt cỏc iu kin ti u cho x lý mu 44 3.2.1 Cỏc phng phỏp x lý mu da phõn tớch cỏc amin thm 44 3.2.2 nh hng ca pH n thu hi ca phng phỏp chit lng lng 45 3.2.3 nh hng ca dung mụi chit 47 3.2.4 Kho sỏt nh hng ca s ln chit 47 3.2.6 Tng hp quy trỡnh x lý mu phõn tớch cỏc amin thm mu da 48 3.3 So sỏnh phng phỏp phõn tớch mi xõy dng vi phng phỏp tiờu chun (TCVN 7536: 2005 ) 54 3.4 ng dng phng phỏp phõn tớch nghiờn cu vo phõn tớch mt s sn phm da ang lu hnh trờn th trng Vit Nam 55 KT LUN 59 TI LIU THAM KHO 60 PH LC 63 PH LC 1: KHO ST IU KIN TI U XC NH CC AMIN THM TRấN DETECTOR MS/MS 64 PH LC 2: NG CHUN XC NH CC AMIN THM BNG H THNG HPLC-MS/MS 83 PH LC 3: QUY TRèNH PHN TCH 18 AMIN THM BNG H THNG HPLC MS/MS 102 DANH MC HèNH V Hỡnh 1: Chuyn húa amin thm t thuc nhum azo .5 Hỡnh 2: thu hi ca cỏc phng phỏp xỏc nh amin thm t mu da tht 19 Hỡnh 1: H thng sc ký lng hiu nng cao kt hp u dũ ph HPLC MS/MS 25 Hỡnh 2: Cỏc thụng s cn ti u húa cho u dũ ph phõn tớch cỏc amin thm 27 Hỡnh 3: Khong tuyn tớnh ca phng phỏp phõn tớch .32 Hỡnh 1: Phõn tỏch sc ký cỏc cht amin thm trờn ct Hypersil Gold C18 34 Hỡnh 2: Phõn tỏch sc ký cỏc cht amin thm trờn ct Hypersil Gold aQ 35 Hỡnh 3: Phõn tỏch sc ký cỏc cht amin thm trờn ct Hypersil Gold PFP 35 Hỡnh 4: Phõn tỏch cỏc cht amin thm trờn ct C18 vi pha ng A-H2O (0,1% FA) v B-MeOH .36 Hỡnh 5: Phõn tỏch cỏc cht amin thm trờn ct C18 vi pha ng AH2O(CH3COONH4 25mM) v B-MeOH 37 Hỡnh 6: Phõn tỏch cỏc cht amin thm trờn ct C18 vi pha ng A-H2O v BMeOH 37 Hỡnh 7: Ti u húa cỏc thụng s cho ngun ion húa ESI .39 Hỡnh 8: Ph tiờu biu ca NTA quột ch Fullscan .40 Hỡnh 9: Ph tiờu biu ca NTA chy ch Fullscan product 40 Hỡnh 10: Ti u húa nng lng bn phỏ ca NTA 41 Hỡnh 11: nh hng ca pH n thu hi (pH=8,93) .46 Hỡnh 12: nh hng ca pH n thu hi (pH=4,13) .46 Hỡnh 13: So sỏnh thu hi s dng cỏc dung mụi chit khỏc .47 Hỡnh 14: nh hng ca s ln chit n thu hi 48 Hỡnh 16: Quy trỡnh x lý mu da phõn tớch cỏc amin thm trờn HPLC-MS/MS 49 Hỡnh 15: : Quy trỡnh x lý mu da phõn tớch cỏc amin thm trờn HPLC-MS/MS 49 Hỡnh 17: chn lc ca phng phỏp phõn tớch i vi cht TDA 50 Hỡnh 18: thu hi ca phng phỏp chit cỏc amin thm 51 Hỡnh 19: ng chun ca ASD khong hm lng 1,0 10 mg/kg 53 DANH MC BNG Bng 1: Danh sỏch cỏc amin thm thuc nhum b EU cm s dng [22] Bng 2: Phõn tớch ỏnh giỏ phn ng ca amin thm 13 Bng 3: thu hi trung bỡnh ca cỏc amin thm quỏ trỡnh kh (n=3) .15 Bng 4: Cỏc phng phỏp phõn tớch xỏc nh thuc nhum azo b cm sn phm tiờu dựng 16 Bng 5: Thụng s ti u cho quỏ trỡnh phõn tớch MS/MS 19 Bng 6: ng chun v gii hn phỏt hin ca cỏc amin (trung bỡnh lp li) 20 Bng 7: thu hi ca hn hp chun amin thm si khụng nhum (trung bỡnh lp li) .21 Bng 1: Thụng s thit lp cho HPLC/DAD 34 Bng 2: Cỏc thụng s thit lp cho ngun ion húa ESI .39 Bng 3: Tng hp mnh ph ca cỏc hp cht amin v nng lng bn phỏ ti u 42 Bng 4: Tng hp thi gian lu ca 21 amin thm 43 Bng 5: Hng s pKa v logKow ca cỏc amin thm 45 Bng 6: lp li ca phng phỏp phõn tớch amin thm da thuc (n=6) 51 Bng 7: ỏnh giỏ gii hn phỏt hin ca phng phỏp phõn tớch da thuc 52 Bng 8: Kt qu tớnh toỏn chm v ỳng theo phng phỏp tiờu chun 54 Bng 9: Kt qu tớnh toỏn chm v ỳng theo phng phỏp nghiờn cu 54 Bng 10: Kt qu phõn tớch thuc nhum azo mu da thuc trờn th trng 56 DANH SCH CC T VIT TT T vit tt TT 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 4-Aminobiphenyl O-Anisidine Atomic Absorption Spectrophotometric Association of Official Agricultural Chemists Benzidine Bureau Veritas Quality International 4-Chloroaniline 4-Chloro-o-toluidine Comitộ Europộen de Normalisation Consumer Product Safety Improvement Act 4-Cresidine Deutsches Institut fỹr Normung Diode Array Detector 4,4-Diaminodiphenylmethane 3,3'-Dimethoxybenzidine 3,3-Dimetylbenzidine 3,3-Dimethyl-4,4 diaminodiphenylmethane 3,3'-Diclobenzidine 2,4-Diaminotoluen Electron Spray Ionization European Economic Community European Commission Gas Chromatography High Performance Liquid Chromatography High Performance Capillary Electrophoresis Inductively Coupled Plasma Mass Spectrometry International Standard Organization International Union of Pure and Applied Chemistry Limit of Detector Limit of Quantitation Mass Spectrography Method Detector Limit 4,4'-Metylen-bis-(2-cloaniline) 2-Naphthylamine 35 Organisation for Economic Co-operation and Development Tờn vit tt ABP ASD AAS AOAC BZD BVQI CAN COT CEN CPSIA CSD DIN DAD DDM DTB DBD DPM DBZ DAT ESI EEC EC GC HPLC HPCE ICP-MS ISO IUPAC LOD LOQ MS MDL MTB NTA OECD TT 36 37 38 39 40 41 42 43 44 45 46 T vit tt 4,4'-Oxydianiline Recovery Standard Deviation Registration Evaluation Authorization Restriction Shoe and Allied Trades Research Association Tiờu chun Vit Nam Thin Layer Chromatography 4,4'-Thiodianiline o-Toluidine 2,4,5-Trimethylaniline Ultravioletvisible spectroscop Vn phũng Cụng nhn Cht lng Tờn vit tt ODA RSD REACH SATRA TCVN TCL TDA TLD TMA UV-Vis BoA Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 4,4_Thiodianiline Y = -3.97799e+006+2.25479e+006*X R^2 = 0.9975 W: 1/X 20000000 18000000 16000000 14000000 Area TDA 12000000 10000000 8000000 6000000 4000000 2000000 0 E:\tai lieu\ \Mix20amin_1mg_kg_2ul mg/kg 10 6/17/2014 5:26:46 PM RT: 15.00 - 25.00 SM: 15G Relative Abundance mg/kg NL: 1.37E4 TIC F: + c ESI SRM ms2 217.000 [123.500-124.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 22.36 AA: 393163 100 80 60 40 RT: 24.15 AA: 557 20 15 16 17 18 19 20 21 22 23 24 25 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul 6/17/2014 6:27:51 PM RT: 15.00 - 25.00 SM: 15G RT: 22.39 AA: 1803171 100 NL: 6.28E4 TIC F: + c ESI SRM ms2 217.000 [123.500-124.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 11 15 16 17 18 19 20 21 22 23 24 25 Time (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 15.00 - 25.00 SM: 15G NL: 2.29E5 RT: 22.37 AA: 6749593 100 TIC F: + c ESI SRM ms2 217.000 [123.500-124.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 23.81 RT: 24.31 AA: 1259 AA: 2177 10 15 16 17 18 19 20 21 22 23 24 25 Time (min) E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul 6/17/2014 8:29:58 PM RT: 15.00 - 25.00 SM: 15G RT: 22.39 AA: 14446009 100 NL: 4.85E5 TIC F: + c ESI SRM ms2 217.000 [123.500-124.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 24.14 AA: 1054 15 16 17 18 19 20 21 22 23 24 25 Time (min) E:\tai lieu\ \Mix20amin_10mg_kg_2ul 6/17/2014 9:31:03 PM RT: 15.00 - 25.00 SM: 15G RT: 22.40 AA: 18464426 100 NL: 6.35E5 TIC F: + c ESI SRM ms2 217.000 [123.500-124.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 16.82 AA: 757 15 16 17 RT: 23.88 RT: 24.97 AA: 643 AA: 4034 RT: 20.43 AA: 1802 18 19 20 21 22 23 24 25 Time (min) 94 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 2-Naphthyamin Y = -202944+1.15958e+006*X R^2 = 0.9940 W: 1/X 12000000 11000000 10000000 9000000 NTA 8000000 Area 7000000 6000000 5000000 4000000 3000000 2000000 1000000 0 E:\tai lieu\ \Mix20amin_1mg_kg_2ul mg/kg 10 6/17/2014 5:26:46 PM RT: 15.00 - 25.00 SM: 15G 100 Relative Abundance mg/kg NL: 3.46E4 TIC F: + c ESI SRM ms2 144.100 [126.500-127.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 22.17 AA: 1084562 80 60 40 RT: 24.07 AA: 2584 20 15 16 17 18 19 20 21 22 23 24 25 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul RT : 15.00 - 25.00 6/17/2014 6:27:51 PM SM: 15G RT : 22.20 AA: 2273343 100 NL: 7.01E4 T IC F: + c ESI SRM ms2 144.100 [126.500-127.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 12 70 60 50 40 30 20 RT : 23.75 AA: 2597 10 15 16 17 18 19 20 21 22 23 RT : 24.84 AA: 8675 24 25 T ime (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 15.00 - 25.00 SM: 15G RT: 22.18 AA: 6775916 100 NL: 2.02E5 TIC F: + c ESI SRM ms2 144.100 [126.500-127.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 24.86 AA: 22434 15 16 17 18 19 20 21 22 23 24 25 Time (min) 6/17/2014 8:29:58 PM E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul RT: 15.00 - 25.00 SM: 15G RT: 22.20 AA: 10203976 100 NL: 3.07E5 TIC F: + c ESI SRM ms2 144.100 [126.500-127.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 23.71 AA: 5419 10 15 16 17 18 E:\tai lieu\ \Mix20amin_10mg_kg_2ul 19 20 21 22 23 24 25 (min) 6/17/2014 Time 9:31:03 PM RT: 15.00 - 25.00 SM: 15G RT: 22.21 AA: 11729068 100 NL: 3.56E5 TIC F: + c ESI SRM ms2 144.100 [126.500-127.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 15 16 17 18 19 20 21 22 23 24 25 Time (min) 95 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 4-clo-2-toluidine Y = 9699.54+126367*X R^2 = 0.9977 W: 1/X 1200000 1000000 COT Area 800000 600000 400000 200000 0 mg/kg E:\tai lieu\ \Mix20amin_1mg_kg_2ul 10 6/17/2014 5:26:46 PM RT: 20.00 - 30.00 SM: 15G Relative Abundance mg/kg NL: 3.63E3 TIC F: + c ESI SRM ms2 142.100 [106.500-107.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 23.57 AA: 136058 100 80 60 40 RT: 25.16 AA: 2620 RT: 21.98 AA: 1472 20 RT: 27.02 AA: 2905 RT: 29.47 AA: 1391 20 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul 6/17/2014 6:27:51 PM RT: 20.00 - 30.00 SM: 15G RT: 23.56 AA: 322670 100 NL: 9.46E3 TIC F: + c ESI SRM ms2 142.100 [106.500-107.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 13 10 RT: 27.68 AA: 4861 RT: 20.14 AA: 1629 20 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 20.00 - 30.00 SM: 15G RT: 23.58 AA: 823132 100 NL: 2.15E4 TIC F: + c ESI SRM ms2 142.100 [106.500-107.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 22.10 AA: 1147 20 21 22 23 24 25 26 27 28 29 30 Time (min) 6/17/2014 8:29:58 PM E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul RT: 20.00 - 30.00 SM: 15G RT: 23.60 AA: 1199215 100 NL: 3.20E4 TIC F: + c ESI SRM ms2 142.100 [106.500-107.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 25.35 AA: 6019 10 RT: 26.82 AA: 2598 20 21 22 23 24 25 26 27 RT: 28.46 AA: 2560 28 RT: 29.97 AA: 5524 29 30 Time (min) E:\tai lieu\ \Mix20amin_10mg_kg_2ul 6/17/2014 9:31:03 PM RT: 20.00 - 30.00 SM: 15G RT: 23.61 AA: 1225320 100 NL: 3.27E4 TIC F: + c ESI SRM ms2 142.100 [106.500-107.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 25.40 AA: 5159 10 RT: 26.76 AA: 2200 20 21 22 23 24 25 26 27 28 29 30 Time (min) 96 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 2,4,5-trimetylaniline Y = 25560.8+361502*X R^2 = 0.9962 W: 1/X 4000000 3500000 3000000 TMA Area 2500000 2000000 1500000 1000000 500000 0 mg/kg E:\tai lieu\ \Mix20amin_1mg_kg_2ul 10 6/17/2014 5:26:46 PM RT: 20.00 - 30.00 SM: 15G Relative Abundance mg/kg NL: 1.30E4 TIC F: + c ESI SRM ms2 136.200 [120.500-121.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 25.82 AA: 396011 100 80 60 40 RT: 21.32 AA: 10448 20 RT: 27.30 AA: 14768 RT: 24.62 AA: 8744 RT: 23.50 AA: 2073 RT: 28.77 AA: 25485 20 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul 6/17/2014 6:27:51 PM RT: 20.00 - 30.00 SM: 15G NL: 2.71E4 RT: 25.81 AA: 893016 100 TIC F: + c ESI SRM ms2 136.200 [120.500-121.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 14 10 RT: 20.22 AA: 7415 RT: 23.52 AA: 24479 RT: 22.43 AA: 16741 RT: 28.42 RT: 28.88 RT: 29.74 AA: 4226 AA: 7294 AA: 15867 RT: 24.76 AA: 8526 20 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 20.00 - 30.00 SM: 15G NL: 5.70E4 RT: 25.83 AA: 1946367 100 TIC F: + c ESI SRM ms2 136.200 [120.500-121.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 21.13 AA: 9015 10 RT: 22.53 AA: 10083 RT: 28.86 RT: 29.32 AA: 9623 AA: 21176 RT: 23.66 AA: 13641 20 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul 6/17/2014 8:29:58 PM RT: 20.00 - 30.00 SM: 15G RT: 25.86 AA: 3042386 100 NL: 8.59E4 TIC F: + c ESI SRM ms2 136.200 [120.500-121.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 20.65 RT: 21.39 AA: 6321 AA: 12560 10 RT: 27.80 AA: 13186 RT: 22.90 AA: 9821 RT: 29.47 AA: 30615 20 21 22 23 24 E:\tai lieu\ \Mix20amin_10mg_kg_2ul 25 26 27 28 29 30 (min) 6/17/2014Time 9:31:03 PM RT: 20.00 - 30.00 SM: 15G RT: 25.83 AA: 3400802 100 NL: 9.94E4 TIC F: + c ESI SRM ms2 136.200 [120.500-121.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 20.94 AA: 10842 10 RT: 27.69 AA: 13960 RT: 23.58 AA: 26929 RT: 22.22 AA: 6595 RT: 29.24 AA: 9003 20 21 22 23 24 25 26 27 28 29 30 Time (min) 97 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 3,3-dimetyl-4,4-diaminodiphenylmetal Y = -2.09639e+006+8.57414e+006*X R^2 = 0.9939 W: 1/X 90000000 80000000 70000000 60000000 50000000 Area DPM 40000000 30000000 20000000 10000000 0 mg/kg E:\tai lieu\ \Mix20amin_1mg_kg_2ul 10 6/17/2014 5:26:46 PM RT: 20.00 - 30.00 SM: 15G Relative Abundance mg/kg NL: 2.44E5 TIC F: + c ESI SRM ms2 227.100 [119.500-120.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 27.92 AA: 7071547 100 80 60 40 RT: 20.36 AA: 1939 20 20 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul 6/17/2014 6:27:51 PM RT: 20.00 - 30.00 SM: 15G RT: 27.95 AA: 16314338 100 NL: 5.56E5 TIC F: + c ESI SRM ms2 227.100 [119.500-120.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 15 2,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 20.26 AA: 9915 20 RT: 26.28 AA: 4729 21 22 23 24 25 26 27 28 29 30 Time (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 20.00 - 30.00 SM: 15G RT: 27.93 AA: 47291546 100 NL: 1.56E6 TIC F: + c ESI SRM ms2 227.100 [119.500-120.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 20.36 AA: 10293 20 RT: 25.75 AA: 2730 21 22 23 24 25 26 27 28 29 30 Time (min) PM 6/17/2014 8:29:58 E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul RT: 20.00 - 30.00 SM: 15G RT: 27.99 AA: 71892791 100 NL: 2.36E6 TIC F: + c ESI SRM ms2 227.100 [119.500-120.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 20.22 AA: 13985 20 RT: 22.24 AA: 5992 21 22 23 E:\tai lieu\ \Mix20amin_10mg_kg_2ul 24 25 26 27 28 29 30 Time (min) 6/17/2014 9:31:03 PM RT: 20.00 - 30.00 SM: 15G RT: 27.96 AA: 80789589 100 NL: 2.69E6 TIC F: + c ESI SRM ms2 227.100 [119.500-120.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 21.33 AA: 2721 20 21 RT: 22.22 AA: 3302 22 RT: 23.97 AA: 3739 23 24 RT: 26.72 AA: 2085 25 26 27 28 29 30 Time (min) 98 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 4-aminobiphenyl Y = -108836+3.54265e+006*X R^2 = 0.9933 W: 1/X 35000000 30000000 25000000 Area ABP 20000000 15000000 10000000 5000000 0 mg/kg E:\tai lieu\ \Mix20amin_1mg_kg_2ul 10 6/17/2014 5:26:46 PM RT: 25.00 - 35.00 SM: 15G Relative Abundance mg/kg NL: 1.16E5 TIC F: + c ESI SRM ms2 170.000 [151.500-152.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 29.78 AA: 3639462 100 80 60 40 20 RT: 25.25 AA: 6682 25 RT: 27.18 AA: 12555 26 27 RT: 32.42 AA: 5013 28 29 30 31 32 RT: 34.05 AA: 7802 33 34 35 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul RT : 25.00 - 35.00 6/17/2014 6:27:51 PM SM: 15G RT : 29.78 AA: 7321541 100 NL: 2.31E5 T IC F: + c ESI SRM ms2 170.000 [151.500-152.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 16 10 RT : 26.87 RT : 27.41 AA: 3950 AA: 3821 25 26 27 RT : 31.18 AA: 2510 RT : 28.34 AA: 5170 28 29 30 31 RT : 32.92 RT : 33.31 AA: 4298 AA: 2025 32 33 RT : 34.24 AA: 5329 34 35 T ime (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 25.00 - 35.00 SM: 15G RT: 29.79 AA: 20189699 100 NL: 6.36E5 TIC F: + c ESI SRM ms2 170.000 [151.500-152.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 26.65 RT: 27.31 AA: 5277 AA: 13079 25 26 27 RT: 32.66 RT: 33.13 AA: 6511 AA: 6153 28 29 30 31 32 33 RT: 34.14 AA: 11455 34 35 Time (min) E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul 6/17/2014 8:29:58 PM RT: 25.00 - 35.00 SM: 15G NL: 9.66E5 RT: 29.82 AA: 31090301 100 TIC F: + c ESI SRM ms2 170.000 [151.500-152.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 26.32 AA: 2407 25 RT: 31.84 RT: 32.38 RT: 33.16 AA: 3583 AA: 3878 AA: 4604 RT: 27.18 AA: 4285 26 27 28 29 30 31 32 33 34 35 Time (min) E:\tai lieu\ \Mix20amin_10mg_kg_2ul 6/17/2014 9:31:03 PM RT: 25.00 - 35.00 SM: 15G RT: 29.79 AA: 33887383 100 NL: 1.06E6 TIC F: + c ESI SRM ms2 170.000 [151.500-152.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 34.48 AA: 12769 25 26 27 28 29 30 31 32 33 34 35 Time (min) 99 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 3,3-diclobenzidine Y = -252827+283925*X R^2 = 0.9909 W: 1/X 2500000 2000000 Area DBZ 1500000 1000000 500000 0 E:\tai lieu\ \Mix20amin_1mg_kg_2ul mg/kg 10 6/17/2014 5:26:46 PM RT: 25.00 - 35.00 SM: 15G Relative Abundance mg/kg NL: 3.98E3 TIC F: + c ESI SRM ms2 253.000 [216.500-217.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 32.34 AA: 117894 100 80 60 40 RT: 26.88 AA: 3958 20 RT: 34.44 AA: 31438 RT: 30.64 AA: 7812 RT: 29.12 AA: 8047 25 26 27 28 29 30 31 32 33 34 35 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul 6/17/2014 6:27:51 PM RT: 25.00 - 35.00 SM: 15G RT: 32.34 AA: 324583 100 NL: 9.96E3 TIC F: + c ESI SRM ms2 253.000 [216.500-217.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 17 70 60 50 40 30 RT: 31.37 AA: 1264 20 RT: 26.48 RT: 27.18 AA: 16468 AA: 1216 RT: 25.24 AA: 10356 10 RT: 28.69 AA: 6090 RT: 29.70 AA: 8654 RT: 34.48 AA: 3147 RT: 30.87 AA: 6426 25 26 27 28 29 30 31 32 33 34 35 Time (min) E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 25.00 - 35.00 SM: 15G RT: 32.36 AA: 1119098 100 NL: 3.49E4 TIC F: + c ESI SRM ms2 253.000 [216.500-217.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 RT: 25.64 AA: 8178 10 RT: 27.23 RT: 27.86 AA: 4952 AA: 5271 RT: 34.37 AA: 2530 RT: 31.00 AA: 8565 RT: 28.94 RT: 29.52 AA: 2783 AA: 6341 25 26 27 28 29 30 31 32 33 34 35 Time (min) E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul RT : 25.00 - 35.00 6/17/2014 8:29:58 PM SM: 15G RT : 32.39 AA: 2141282 100 NL: 6.55E4 T IC F: + c ESI SRM ms2 253.000 [216.500-217.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT : 25.28 AA: 14762 RT : 27.03 RT : 27.45 AA: 2248 AA: 4254 25 26 27 RT : 29.40 RT : 29.86 AA: 1656 AA: 3245 28 29 30 RT : 34.17 AA: 5001 RT : 31.26 AA: 2304 31 32 33 34 35 T ime (min) E:\tai lieu\ \Mix20amin_10mg_kg_2ul 6/17/2014 9:31:03 PM RT: 25.00 - 35.00 SM: 15G RT: 32.35 AA: 3017015 100 NL: 9.37E4 TIC F: + c ESI SRM ms2 253.000 [216.500-217.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 25.17 AA: 11563 25 RT: 29.60 AA: 4617 RT: 28.32 AA: 7882 26 27 28 29 RT: 34.02 RT: 34.91 AA: 5981 AA: 5176 RT: 30.41 AA: 3964 30 31 32 33 34 35 Time (min) 100 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn STT Amin thm Khoa Húa hc ng chun v khong tuyn tớnh 4,4-metylen-bis-(2-cloaniline) Y = 262165+1.1597e+006*X R^2 = 0.9947 W: 1/X 12000000 10000000 MTB Area 8000000 6000000 4000000 2000000 0 mg/kg E:\tai lieu\ \Mix20amin_1mg_kg_2ul 10 6/17/2014 5:26:46 PM RT: 30.00 - 40.00 SM: 15G Relative Abundance mg/kg NL: 4.55E4 TIC F: + c ESI SRM ms2 267.000 [230.500-231.500] M S ICIS M ix20amin_1mg_kg_2ul RT: 34.36 AA: 1407693 100 80 60 40 RT: 36.46 RT: 37.19 AA: 4192 AA: 2002 RT: 30.48 AA: 4791 20 30 31 32 33 34 35 36 37 38 39 40 Time (min) E:\tai lieu\ \Mix20amin_2,5mg_kg_2ul RT : 30.00 - 40.00 6/17/2014 6:27:51 PM SM: 15G RT : 34.40 AA: 2951984 100 NL: 9.65E4 T IC F: + c ESI SRM ms2 267.000 [230.500-231.500] MS ICIS Mix20amin_2,5mg_k g_2ul 90 2,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT : 35.84 AA: 2279 RT : 32.19 AA: 2189 30 31 32 33 34 35 RT : 39.61 AA: 4012 36 37 38 39 40 T ime (min) 18 E:\tai lieu\ \Mix20amin_5mg_kg_2ul 6/17/2014 7:28:53 PM RT: 30.00 - 40.00 SM: 15G NL: 2.32E5 RT: 34.37 AA: 7282854 100 TIC F: + c ESI SRM ms2 267.000 [230.500-231.500] MS ICIS Mix20amin_5mg_kg _2ul 90 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 33.09 AA: 6346 30 31 32 33 RT: 36.66 RT: 37.44 AA: 1821 AA: 2019 34 35 36 37 38 39 40 Time (min) E:\tai lieu\ \Mix20amin_7,5mg_kg_2ul 6/17/2014 8:29:58 PM RT: 30.00 - 40.00 SM: 15G RT: 34.41 AA: 9321921 100 NL: 3.00E5 TIC F: + c ESI SRM ms2 267.000 [230.500-231.500] MS ICIS Mix20amin_7,5mg_k g_2ul 90 7,5 mg/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 35.81 AA: 2870 RT: 30.68 AA: 2846 30 31 32 33 34 35 36 RT: 38.64 AA: 8119 37 38 39 40 Time (min) E:\tai lieu\ \Mix20amin_10mg_kg_2ul 6/17/2014 9:31:03 PM RT: 30.00 - 40.00 SM: 15G NL: 3.56E5 RT: 34.41 AA: 11051805 100 TIC F: + c ESI SRM ms2 267.000 [230.500-231.500] MS ICIS Mix20amin_10mg_k g_2ul 90 10 g/kg Relative Abundance 80 70 60 50 40 30 20 10 RT: 32.51 AA: 4778 30 31 32 RT: 39.46 AA: 6137 33 34 35 36 37 38 39 40 Time (min) 101 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn Khoa Húa hc PH LC 3: QUY TRèNH PHN TCH 18 AMIN THM BNG H THNG HPLC MS/MS 102 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn Khoa Húa hc PHM VI NG DNG Hng dn ny quy nh phng phỏp xỏc nh cỏc amin thm ó bit cú th gii phúng t thuc nhum azo mu da Gii hn phỏt hin ca phng phỏp l 0,5 mg/Kg mu da NGUYấN TC PHNG PHP Sau loi du, m v cỏc cht chng thm bng n-hexan, thuc nhum azo mu da c chit bng dung dch m citrat (pH=6,0) 70oC bỡnh kớn Cỏc cht amin thm (Bng 1) c gii phúng t thuc nhum azo bng phn ng kh vi natri ithionit Cỏc cht amin thm sau ú c tỏch dung dch chit bng phng phỏp chit lng lng s dng methyl-t-butyl ether (MTBE) Tin hnh phõn tớch xỏc nh cỏc amin bng thit b sc ký lng u dũ ph kộp HPLC-MS/MS Bng Danh sỏch cỏc cht chun amin thm STT 10 11 12 13 14 15 16 17 18 Tờn amin 4-Aminobiphenyl Benzidine 4-Chloro-o-toluidine 2-Naphthylamine 4-Chloroaniline 2,4-Diaminoanisol 4,4-Diaminodiphenylmetan 3,3-Dichlorobenzidine 3,3-Dimetoxybenzidine 3,3-Dimethylbezidine 3,3-Dimethyl-4,4-diaminodiphenylmetan 4-Cresidine 4,4-Methyl-bis-(2-chloroaniline) 4,4-Oxydianiline 4,4-Thiodianiline 2-Toluidine 2,4-Diamonitoluene 2,4,5-Trimethyl anilin S CAS 92-67-1 92-87-5 95-69-2 91-59-8 106-47-8 615-05-4 101-77-9 91-94-1 119-90-4 119-93-7 838-88-0 120-71-8 101-14-4 101-80-4 139-65-1 95-53-4 95-80-7 137-17-7 THIT B V HO CHT 3.1 Thit b - Sc ký lng hiu nng cao: HPLC/DAD Dionex Ultimate 3000 ghộp ni u dũ ph kộp Thermo TSQ Quantum Access Max - Thit b lc cú n nh nhit: GynomaxTM 929 Amerex Instrument, Inc 103 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn Khoa Húa hc - B siờu õm cú iu chnh nhit: Elmasonic S300H - Ct chit lng lng: Chromabond XTR (70 mL, 14,5 g Kieselgur) - H thi khớ nit: tinh khit 99,99% - Mng lc mu: mng nylon 0,45àm - Micropipet 20 200 àL v 100 1000 àL - Pipet 1mL, 2mL, 5mL, 10mL, 25mL - Bỡnh nh mc vi cỏc th tớch khỏc - Bỡnh phn ng bng thy tinh chu nhit, kớn khớ 3.2 Hoỏ cht - Dung dch chun 18 amin thm (Bng 1) - Metanol (MeOH) HPLC grade - n-hexan HPLC grade - Acetonitrile (ACN) HPLC grade - Methyl-t-butyl ether (MTBE) Analytical grade - Amoni ihirophotphat (NaH2PO4) ( tinh khit >99,0%) - Natri hirophotphat (Na2HPO4) ( tinh khit >99,0%) - Natri hiroxit (NaOH) ( tinh khit >96%) - Axit citric ( tinh khit >99,5%) - Axit formic (FA) ( tinh khit 98%) - Dung dch natri ithionit 200 mg/mL, chun b hng ngy (hoỏ cht k thut-Labosi) - Dung dch m citrat (pH=6): 12,526 g axit citric v 6,32 g natri hiroxit, nh mc n 1000 mL bng nc ct - Chlorobenzene ( tinh khit >99,0%) - Nc ct deion LY MU V CHUN B MU Nu cú th ly mu th phự hp vi TCVN 7117 v nghin da theo TCVN 7126: 2002 Nu khụng ly mu theo TCVN 7117 (vớ d nh da ly t sn phm ó hon thin nh giy ng, qun ỏo) thỡ chi tit ca quỏ trỡnh ly mu phi c ghi bỏo cỏo th nghim i vi quỏ trỡnh phõn tớch, mt mu i din gm 1,0 g ( 0,10 g) da ó nghin (kớch thc mm 0,5 mm) c cõn chớnh xỏc cho vo bỡnh phn ng CCH TIN HNH 6.1 X lý mu Bc 1: Loi cht bộo 104 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn Khoa Húa hc Cõn 1,0 g mu da (chớnh xỏc n 0,10 g) ó nghin vo bỡnh kớn 50 ml, thờm 20 ml n-hexan v siờu õm 40 oC 20 phỳt Lp n-hexan sau ú c gn mu da Mu da c x lý nh trờn vi hai ln 20 ml n-hexan Sau gn b dung mụi nhexan, mu da c loi b hon ton n-hexan bng cỏch bay hi t nhiờn qua ờm bỡnh m Bc 2: Tỏch, kh Cho 17 mL dung dch m citrat ó c un núng trc n 70oC vo mu th Bỡnh phn ng c y kớn v lc b iu nhit 25 phỳt 70oC Sau ú thờm ml natri ithionit 200 mg/mL v gi bỡnh phn ng 20 phỳt 70oC kh thuc nhum azo sinh cỏc amin thm Sau quỏ trỡnh kh, mu c lm ngui n nhit phũng b nc ngui Bc 3: Chit lng - lng Dung dch mu sau tỏch kh c gn vo ng ly tõm 50mL v cho thờm 1mL NaOH 1M v g NaCl dung dch cú mụi trng kim Cho thờm 10 ml MTBE v lc mnh hai pha trn ln vo Sau ú, mu c ly tõm 10 phỳt 4500 vũng/phỳt phõn lp Hỳt lp MTBE trờn vo ng nh mc thu tinh Cụ cn dung mụi MTBE n hon ton bng dũng khớ nit Thờm vo 500 àL methanol v 500 àL H2O hũa tan cn Lc dung dch mu cui cựng qua mng lc 0,45 àm v bm àL vo h thng HPLC-MS/MS xỏc nh hm lng cỏc amin thm 6.2 Kim tra h thng phõn tớch kim tra thu hi ca quỏ trỡnh phõn tớch, cho 1,0 ml dung dch chun nng 30 mg/L vo bỡnh phn ng cú cha 17 ml dung dch m citrate (ó un núng n 700C) v 1,0 g ( 0,10 g) mu da trng Tin hnh xỏc nh hm lng cỏc amin thm nh quy trỡnh trờn (5.1) T l thu hi t c phi ớt nht l 20% i vi 2,4Diaminoanisol, 50% i vi 2-Toluidine v 2,4-Diaminotoluene, cỏc amin cũn li t l thu hi t ớt nht 70% 6.3 Hiu chun Nng cỏc cht amin mu th nghim c xỏc nh bng phng phỏp ng chun (xem 6.4) 6.4 Xõy dng ng chun ng chun trờn nn mu da trng c dng khong nng 1,0 10,0 mg/Kg chy trờn HPLC-MS/MS 105 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn Khoa Húa hc Cõn 05 mu da trng vi lng 1,0 g ( 0,10 g) vo 05 bỡnh tam giỏc 50 mL Tin hnh loi m theo bc (5.1) Thờm 17 mL m citrate ó c un núng n 70 0C vo bỡnh thu tinh Thờm vo dung dch chun hn hp cỏc amin thm (xem 3.2) vi lng nh Bng (phõn tớch trờn HPLC-DAD) v Bng (phõn tớch trờn HPLCMS/MS) ng chun c xõy dng da trờn tng quan gia din tớch pic v hm lng ca cỏc amin thm Bng Xõy dng ng chun trờn mu da trng bng HPLC-MS/MS im chun Dung dch chun hn hp 50 mg/L 50 mg/L 50 mg/L 50 mg/L 50 mg/L Lng thờm chun 200 àL 150 àL 100 àL 50 àL 25 àL Lng cõn mu da 1,0 g 1,0 g 1,0 g 1,0 g 1,0 g Nng amin thm mu da blank 10 mg/Kg 30 mg/Kg mg/Kg 2,5 mg/Kg mg/Kg 6.5 Phõn tớch nh tớnh bng sc ký lng hai ln ph HPLC/MS-MS Dung dch mu cui cựng c pha loóng vi t l 1:10 bng metanol v bm 5àL vo h thng HPLC-MS/MS xỏc nh cỏc amin thm cú mt mu Cỏc thụng s chy thit b HPLC-MS/MS c thit lp nh Bng 5: Bng 5: Thụng s thit lp cho HPLC-MS/MS Dung mụi gii 1: Dung mụi gii 2: Ct sc ký: Nhit lũ ct : Tc dũng: Gradient: MeOH H2 O Hypersil Gold C18 150mm x 2,1mm, 3àm 40 oC 0,25 ml/phỳt Bt u 10% dung dch gii 1, tng n 80% dung dch gii 45 phỳt Th tớch bm mu: àl Ngun ion hoỏ: Spray voltage = 4500V Sheath gas = 45 psi Thit lp thụng s cho detector Auxiliary gas = psi hai ln ph MS/MS: Capillary temp = 2700C Vaporized temp = 2500C Skimmer offset = -12 V Tube lens offset = 90 V Ion m Ion Nng TT Amin thm (m/z) (m/z) lng (V) 2,4-Diaminoanisol 173,3 141 10 106 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn 10 11 12 13 14 15 16 17 18 2,4-Diaminotoluene o-Toluidine Benzidin 4,4'-Oxydianiline 4-Chloroaniline 4-Cresidine 4.4-Diaminodiphenylmetan 3,3'-Dimethoxybenzidine 3,3-Dimetylbenzidine 4,4'-Thiodianiline 2-Naphtyamine 4-Chloro-2-toluidine 2,4,5-Trimetylaniline 3,3-Dimetyl-4,4'-diaminodiphenylmetan 4-Aminobiphenyl 3,3'-Dichlorobenzidine 4,4'-methylene-bis-(2-chloroaniline) 123,1 108,1 185,4 201,1 128,1 138,1 199,1 245,0 213,1 217 144,1 142,1 136,2 227,1 170,0 253,0 267,0 Khoa Húa hc 108 91 168 108 93 123 106 230 196 124 127 107 121 120 152 217 231 18 17 17 19 17 14 23 18 18 19 22 16 14 26 29 19 20 Tớnh toỏn kt qu Hm lng ca cỏc amin thm mu da c xỏc nh bng phng phỏp ng chun Nng ca cỏc amin thm cú mu da c xỏc nh theo cụng thc sau: Trong ú: Cmu : Hm lng amin mu da (mg/Kg) Co : Hm lng amin thm o c trờn mu (mg/Kg) Cblank : Hm lng amin thm o c trờn mu blank (mg/Kg) N : H s pha loóng (nu cú) 7.1 m bo kt qu th nghim H s hi qui tuyn tớnh xõy dng ng hiu chun R2 0,99 Tt c cỏc ln o u phi kim tra bng mu trng 7.2 Bỏo cỏo kt qu th nghim Bỏo cỏo kt qu cn cú cỏc thụng tin sau: a) Vin dn tiờu chun TCVN 7536:2005/ISO 17234:2003 b) Loi, ngun gc v mụ t v mu phõn tớch hoc mt phn ca mu 107 Lun Thc s T Th Ngc nh Trng i hc Khoa hc T nhiờn c) Khoa Húa hc Bt k sai lch no so vi quy trỡnh phõn tớch, cỏc bc tin hnh c bit thờm vo d) Kt qu phõn tớch tớnh theo mg/Kg i vi tng cht amin thm c lit kờ riờng bit v bỏo cỏo da vo cỏc giỏ tr ngng sau: Trong trng hp giỏ tr ca cỏc amin < 30 mg/Kg: Theo phõn tớch ó tin hnh, thuc nhum azo gii phúng cỏc amin thm ó lit kờ khụng phỏt hin c Trong trng hp giỏ tr ca mt amin > 30 mg/Kg: kt qu phõn tớch cho thy mu da a xem xột ó c sn xut hoc x lý cú s dng thuc nhum azo cú gii phúng mt hoc nhiu loi amin ó lit kờ Trong trng hp mc ca 4-aminodiphenyl v/hoc 2-napthylamine > 30 mg/Kg: s dng phng phỏp ny ó phỏt hin 4-aminoipheny v/hoc 2-napthyamin Theo nhng hiu bit hin khụng th khng nh chc chn rng ó s dng thuc nhum azo cú gii phúng cỏc amin ó lit kờ khụng cú thờm thụng tin 108 Lun Thc s T Th Ngc nh ... TỰ NHIÊN - TẠ THỊ NGỌC ÁNH NGHIÊN CỨU PHÂN TÍCH MỘT SỐ AMIN THƠM TRONG CÁC SẢN PHẨM DA BẰNG PHƢƠNG PHÁP LC/MS/MS CHUYÊN NGÀNH: HÓA PHÂN TÍCH MÃ SỐ: 60440118 LUẬN VĂN THẠC SĨ KHOA HỌC... mẫu phân tích amin thơm mẫu da 48 3.3 So sánh phƣơng pháp phân tích xây dựng với phƣơng pháp tiêu chuẩn (TCVN 7536: 2005 ) 54 3.4 Ứng dụng phƣơng pháp phân tích nghiên cứu vào phân. .. 21 amin thơm 43 Bảng 5: Hằng số pKa logKow amin thơm 45 Bảng 6: Độ lặp lại phƣơng pháp phân tích amin thơm da thuộc (n=6) 51 Bảng 7: Đánh giá giới hạn phát phƣơng pháp phân tích da