I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - - TRN NG TUN NGHIấN CU XC NH OXALAT TRONG MU THC PHM BNG PHNG PHP IN DI MAO QUN S DNG DETECTOR O DN KHễNG TIP XC LUN VN THC S KHOA HC H Ni I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - - TRN NG TUN NGHIấN CU XC NH OXALAT TRONG MU THC PHM BNG PHNG PHP IN DI MAO QUN S DNG DETECTOR O DN KHễNG TIP XC Chuyờn ngnh: Húa phõn tớch Mó s: 60440118 LUN VN THC S KHOA HC NGI HNG DN KHOA HC: TS NGUYN TH NH HNG H Ni LI CM N Sau mt thi gian nghiờn cu v hc tụi ó hon thnh lun cao hc ca mỡnh vi ti: Nghiờn cu xỏc nh oxalate mu thc phm bng phng phỏp in di mao qun s dng detector o dn khụng tip xỳc di s hng dn ca TS Nguyn Th nh Hng v cỏc thy cụ, anh ch, cỏc bn b mụn Húa phõn tớch Vi lũng kớnh trng v bit n sõu sc tụi xin chõn thnh cm n TS Nguyn Th nh Hng ngi ó giao ti v tn tỡnh ch dn tụi quỏ trỡnh hon thnh lun Tụi xin chõn thnh cm n Cụng ty 3Sanalysis v i tỏc ti Khoa Húa hc, Trng i hc Basel, Thy s ó thit k lp t v h tr cỏc trang thit b cng nh t k thut quỏ trỡnh thc hin nghiờn cu ny Tụi xin chõn thnh cm n cỏc Thy cụ B mụn Húa Phõn tớch núi riờng v khoa Húa hc núi chung ó dy d, ch bo v ng viờn tụi thi gian hc ti trng i hc Khoa hc T nhiờn H Ni Tụi xin cm n sinh viờn Nguyn Th Minh Hũa, lp k57T Ti nng Húa, sinh viờn Nguyn Th Liờn, lp K56A Húa hc ó h tr tụi quỏ trỡnh nghiờn cu Cui cựng, tụi xin gi li cm n gia ỡnh, cỏc bn hc viờn v sinh viờn b mụn Húa phõn tớch ó giỳp tụi thi gian hc v nghiờn cu ny Hc viờn Trn ng Tun MC LC M U CHNG 1: TNG QUAN in di, tc in di v thi gian in di .14 1.6.1.Mao qun 15 1.6.2.Dung dch m pH v pha ng phng phỏp in di mao qun .17 1.6.3.Ngun in th cao .17 1.6.4.K thut bm mu phng phỏp in di mao qun 18 1.7.Phng phỏp in di mao qun s dng detector o dn khụng tip xỳc (CE - C4D) 19 CHNG 2: THC NGHIM 21 CHNG 3: KT QU V THO LUN 27 chm ca thit b 40 3.3.5.Phõn tớch i chng phng phỏp CE-C4D vi phng phỏp HPLC 50 DANH MC TI LIU THAM KHO 52 Ting vit 52 1.Nguyn Th nh Hng (2010), Nghiờn cu xỏc nh cỏc dng asen vụ c nc ngm bng phng phỏp in di mao qun s dng detector dn khụng tip xỳc, Lun ỏn Tin s Húa hc, Trng i hc Khoa hc T nhiờn - HQGHN 52 2.o M Thanh (2012), Ph gia thc phm v sc khe ngi tiờu dựng, Trung tõm Y t D phũng TP.HCM 52 3.T Th Tho (2010), Bi ging chuyờn thng kờ húa phõn tớch, H Quc gia H Ni 52 4.Nguyn Duy Thnh(2004), Cỏc cht ph gia s dng thc phm, bi ging cho hc viờn cao hc chuyờn ngnh cụng nghip thc phm, H Bỏch khoa H Ni 52 5.Vin kim nghim ATVSTP Quc Gia (2010), Thm nh phng phỏp phõn tớch húa hc v vi sinh vt, Nh xut bn khoa hc k thut H Ni .52 6.Ali A Ensafi, S Abbasi, B Rezaei (2001) Kinetic spectrophotometric method for the determination of oxalic acid by its catalytic effect on the oxidation of safranine by dichromate, Spectrochimica Acta Part A, (57), 1833 1838 52 7.B.G Wolthers, W Koolstra, M Hayer, H Elzinga (1987), Development of a reference method for determining urinary oxalate by means of isotope dilutionmass spectrometry (ID-MS) and its usefulness in testing existing assays for urinary oxalate, Clinica Chimica Acta, 170:227-235 .52 8.Corinne Rivasseau , Anne-Marie Boisson, Gaă elle Mong elard, Georgy Couram, Olivier Bastien, Richard Bligny (2006) Rapid analysis of organic acid in plant extracts by capillary electrophoresis with indirect UV detection Directed metabolic analyses during metal stress, Journal of Chromatography A, (112), 283290 .52 9.Curhan (1999), GC epidemiologische Hinweise fỹr die Rolle des Oxalat- Nierensteine spontan, J Endourol 13 (9): 629-31 .53 10.Hoppe B, Kemper MJ, Hvizd MG, Sailer DE, Langman CB (1998), Simultaneous determination of oxalate, citrate and sulfate in children's plasma with ion chromatography, Kidney Int, 53(5):1348-52 53 11.Mariỏn Masỏr et al (2003), Determination of oxalate in beer by zone electrophoresis on a chip with conductivity detection, Journal of seperation science, 26, 647-652 53 12.Michael Trevaskis, V Craige Trenerry (1996) An investigation into the determination of oxalic acid in vegetables by capillary electrophoresis, Food Chemisfry, (57), 323-330 53 13.Muhammad Iqbal Bhanger, M Hassan Khaskhali, F D Khand (1996), Simultaneous Determination of Oxalic acid and Citric acid in Urine by High-erformance Liquid Chromatogrphy, Journal of chromatography.B, Biomedical applications, 675(1), 147-51 53 14.M Hassan Khaskhali, M Iqbal Bhanger, F.D Khand (1996) Simultaneous determination of oxalic and citric acid in urine by high-performance liquid chromatography, Journal of Chromatography B, (675), 147-151 53 15.Nguyen Thi Anh Huong et al (2014), Simple semi-automated portable capillary electrophoresis instrument with contactless conductivity detection for the determination of agonists in pharmaceutical and pig-feed samples, Journal of Chromatography A, Vol 1360, pp 305-311 53 16.Nguyen Thi Anh Huong, Pavel Kubỏn, Viet Hung Pham, Peter C Hauser (2007), Study of the determination of inorganic arsenic species by CE with capacitively coupled contactless conductivity detection, Electrophoresis 2007, (28), 35003506 53 17.Petr Kubỏet al (2014), Separation of oxalate, formate and glycolate in human body fluid samples by capillary electrophoresis with contactless conductometric detection, Journal of Chromatography A, 1325, 24124 .53 18.Ph Puig, M Arellano, J Andrianary, F Dedieu, F Couderc (1997), Method development and validation for the simultaneous determination of organic and inorganic acid by capillary zone electrophoresis, Journal of Chromatography A, 765, 321328 53 19.Ruben Rellan-Alvarez (2011), Development of a new high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry method for the determination of low molecular mass organic acid in plant tissue extracts, J.Agric Food Chem, 59, 6864-6870 53 20.United States Department of Agriculture (1984), Informationsdienst Human Nutrition, Landwirtschaftliches Handbuch Nr 8-11 54 21.Verúnica Galli, Coral Barbas (2004), Capillary electrophoresis for the analysis of short-chain organic acid in coffee, Journal of Chromatography A, 1032: 299304 .54 22.Xiumei Geng , Sufang Zhang , Qian Wang , Zongbao (Kent) Zhao (2008), Determination of organic acid in the presence of inorganic anions by ion chromatography with suppresse d conductivity detection, Journal of Chromatography A, 1192 : 187190 .54 23.Fengwu Wu, Zhike He, Qingyao Luo, Yun'e Zeng (1999), HPLC determination of oxalic acid using tris(1,10-phenanthroline) ruthenium (II) chemiluminescence application to the analysis of spinach, Food Chemistry, (65),543-546 54 25 J G March, B M Simonet, F Grỏses, J A Muủoz, M Valiente (2003), Determination of trace amounts of oxalate in renal calculi and related samples by gas chromatography-mass spectrometry, Journal of Chromatography A,57(11), 811-817 54 DANH MC CC BNG M U CHNG 1: TNG QUAN Bng 1.1 Thụng tin chung v hp cht oxalic, tartaric v citric in di, tc in di v thi gian in di .14 1.6.1.Mao qun 15 1.6.2.Dung dch m pH v pha ng phng phỏp in di mao qun .17 Bng 1.2 Cỏc cht thng dựng lm pha ng CE v giỏ tr pK ca chỳng 17 1.6.3.Ngun in th cao .17 1.6.4.K thut bm mu phng phỏp in di mao qun 18 1.7.Phng phỏp in di mao qun s dng detector o dn khụng tip xỳc (CE - C4D) 19 CHNG 2: THC NGHIM 21 CHNG 3: KT QU V THO LUN 27 Bng 3.1 Kt qu kho sỏt nh hng ca pH n din tớch pic (Spic) v thi gian di chuyn (tdc) ca oxalate, tartrate v citrate 30 Bng 3.2 Kt qu kho sỏt nh hng ca th in di n din tớch pic (Spic) v thi gian di chuyn (tdc) ca oxalate, tartrate v citrate 34 Bng 3.3 S ph thuc din tớch pic vo nng oxalate, tartrate, citrate 36 Bng 3.4 Phng trỡnh ng chun ca oxalate, tartrate v citrate 37 Bng 3.5 Kt qu so sỏnh gia giỏ tr a vi giỏ tr ca phng trỡnh ng chun oxalate, tartrate, citrate 38 Bng 3.6 Gii hn phỏt hin oxalate, tartrate v citrate bng phng phỏp in di mao qun CEC4D 39 Bng 3.7 Giỏ tr khong tuyn tớnh v LOD, LOQ ca oxalate, tartrate, citrate .40 chm ca thit b 40 Bng 3.8 Kt qu xỏc nh lp li ca phng phỏp CE-C4D nh lng oxalate 40 Bng 3.9 Kt qu xỏc nh lp li ca phng phỏp CE-C4D nh lng tartrate 40 Bng 3.10 Kt qu xỏc nh lp li ca phng phỏp CE-C4D nh lng citrate 41 Bng 3.11 Kt qu kho sỏt ỳng ca phng phỏp da trờn thờm chun oxalate 42 Bng 3.12 Kt qu kho sỏt ỳng ca phng phỏp da trờn thờm chun tartrate .42 Bng 3.13 Kt qu kho sỏt ỳng ca phng phỏp da trờn thờm chun citrate 43 Bng 3.14 Thụng tin v c im ca tng loi bia .43 Bng 3.15 Kt qu xỏc nh hm lng cỏc cht oxalate, citrate mt s mu bia .47 3.3.5.Phõn tớch i chng phng phỏp CE-C4D vi phng phỏp HPLC 50 Bng 3.16 So sỏnh kt qu phõn tớch i chng hm lng oxalate, citrate v tartrate mu m tụm 50 DANH MC TI LIU THAM KHO 52 Ting vit 52 1.Nguyn Th nh Hng (2010), Nghiờn cu xỏc nh cỏc dng asen vụ c nc ngm bng phng phỏp in di mao qun s dng detector dn khụng tip xỳc, Lun ỏn Tin s Húa hc, Trng i hc Khoa hc T nhiờn - HQGHN 52 2.o M Thanh (2012), Ph gia thc phm v sc khe ngi tiờu dựng, Trung tõm Y t D phũng TP.HCM 52 3.T Th Tho (2010), Bi ging chuyờn thng kờ húa phõn tớch, H Quc gia H Ni 52 4.Nguyn Duy Thnh(2004), Cỏc cht ph gia s dng thc phm, bi ging cho hc viờn cao hc chuyờn ngnh cụng nghip thc phm, H Bỏch khoa H Ni 52 5.Vin kim nghim ATVSTP Quc Gia (2010), Thm nh phng phỏp phõn tớch húa hc v vi sinh vt, Nh xut bn khoa hc k thut H Ni .52 6.Ali A Ensafi, S Abbasi, B Rezaei (2001) Kinetic spectrophotometric method for the determination of oxalic acid by its catalytic effect on the oxidation of safranine by dichromate, Spectrochimica Acta Part A, (57), 1833 1838 52 7.B.G Wolthers, W Koolstra, M Hayer, H Elzinga (1987), Development of a reference method for determining urinary oxalate by means of isotope dilutionmass spectrometry (ID-MS) and its usefulness in testing existing assays for urinary oxalate, Clinica Chimica Acta, 170:227-235 .52 8.Corinne Rivasseau , Anne-Marie Boisson, Gaă elle Mong elard, Georgy Couram, Olivier Bastien, Richard Bligny (2006) Rapid analysis of organic acid in plant extracts by capillary electrophoresis with indirect UV detection Directed metabolic analyses during metal stress, Journal of Chromatography A, (112), 283290 .52 9.Curhan (1999), GC epidemiologische Hinweise fỹr die Rolle des Oxalat- Nierensteine spontan, J Endourol 13 (9): 629-31 .53 10.Hoppe B, Kemper MJ, Hvizd MG, Sailer DE, Langman CB (1998), Simultaneous determination of oxalate, citrate and sulfate in children's plasma with ion chromatography, Kidney Int, 53(5):1348-52 53 11.Mariỏn Masỏr et al (2003), Determination of oxalate in beer by zone electrophoresis on a chip with conductivity detection, Journal of seperation science, 26, 647-652 53 12.Michael Trevaskis, V Craige Trenerry (1996) An investigation into the determination of oxalic acid in vegetables by capillary electrophoresis, Food Chemisfry, (57), 323-330 53 13.Muhammad Iqbal Bhanger, M Hassan Khaskhali, F D Khand (1996), Simultaneous Determination of Oxalic acid and Citric acid in Urine by High-erformance Liquid Chromatogrphy, Journal of chromatography.B, Biomedical applications, 675(1), 147-51 53 14.M Hassan Khaskhali, M Iqbal Bhanger, F.D Khand (1996) Simultaneous determination of oxalic and citric acid in urine by high-performance liquid chromatography, Journal of Chromatography B, (675), 147-151 53 15.Nguyen Thi Anh Huong et al (2014), Simple semi-automated portable capillary electrophoresis instrument with contactless conductivity detection for the determination of agonists in pharmaceutical and pig-feed samples, Journal of Chromatography A, Vol 1360, pp 305-311 53 16.Nguyen Thi Anh Huong, Pavel Kubỏn, Viet Hung Pham, Peter C Hauser (2007), Study of the determination of inorganic arsenic species by CE with capacitively coupled contactless conductivity detection, Electrophoresis 2007, (28), 35003506 53 17.Petr Kubỏet al (2014), Separation of oxalate, formate and glycolate in human body fluid samples by capillary electrophoresis with contactless conductometric detection, Journal of Chromatography A, 1325, 24124 .53 18.Ph Puig, M Arellano, J Andrianary, F Dedieu, F Couderc (1997), Method development and validation for the simultaneous determination of organic and inorganic acid by capillary zone electrophoresis, Journal of Chromatography A, 765, 321328 53 19.Ruben Rellan-Alvarez (2011), Development of a new high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry method for the determination of low molecular mass organic acid in plant tissue extracts, J.Agric Food Chem, 59, 6864-6870 53 20.United States Department of Agriculture (1984), Informationsdienst Human Nutrition, Landwirtschaftliches Handbuch Nr 8-11 54 21.Verúnica Galli, Coral Barbas (2004), Capillary electrophoresis for the analysis of short-chain organic acid in coffee, Journal of Chromatography A, 1032: 299304 .54 22.Xiumei Geng , Sufang Zhang , Qian Wang , Zongbao (Kent) Zhao (2008), Determination of organic acid in the presence of inorganic anions by ion chromatography with suppresse d conductivity detection, Journal of Chromatography A, 1192 : 187190 .54 23.Fengwu Wu, Zhike He, Qingyao Luo, Yun'e Zeng (1999), HPLC determination of oxalic acid using tris(1,10-phenanthroline) ruthenium (II) chemiluminescence application to the analysis of spinach, Food Chemistry, (65),543-546 54 25 J G March, B M Simonet, F Grỏses, J A Muủoz, M Valiente (2003), Determination of trace amounts of oxalate in renal calculi and related samples by gas chromatography-mass spectrometry, Journal of Chromatography A,57(11), 811-817 54 25,3 142 240 24,6 146 231 25,5 145 25,2 2,26 144 1,68 Spic trung bỡnh RSD (%) 240 237 1,56 Bng 3.10 Kt qu xỏc nh lp li ca phng phỏp CE-C4D nh lng citrate Ln bm mu Din tớch pic cỏc mc nng ca citrate 1,0.10 M 5,0.10-5M 7,5.10-5M 21,9 169 275 -5 20,8 166 277 22,1 169 275 21,0 166 290 21,2 167 270 21,1 171 281 Spic trung bỡnh RSD (%) 21,4 2,46 168 1,19 278 2,47 T bng kt qu cho thy, c mc nng u cú lch chun tng i (RSD) < 3% nm gii hn cho phộp c hm lng ppm Nh vy phng phỏp cú lp li tt ỳng ca phng phỏp Ngoi lp li thỡ ỳng l mt yu t ỏnh giỏ phng phỏp phõn tớch ỳng ca phng phỏp c ỏnh giỏ qua hiu sut thu hi ca phng phỏp phõn tớch, bao gm c x lý v o mu phõn tớch Nú cho bit lng cht b mt i quỏ trỡnh x lý mu ỏnh giỏ hiu sut thu hi l ỏnh giỏ tin cy ca phng phỏp x lý mu ó la chn Trong cỏc thc phm núi chung v cỏc mu thc phm c la chn thng cú thnh phn húa hc phc tp, rt khú cú th tỡm c mu trng cú thnh phn 41 nn ging nh mu thc nhng khụng cha cht phõn tớch Vỡ vy, ỏnh giỏ thu hi ca phng phỏp, chỳng tụi s dng phng phỏp thờm chun, hn hp cht chun oxalate, tartrate v citrate c thờm vo nn mu th l mu bia chai H Ni Quỏ trỡnh ỏnh giỏ c thc hin mc thờm chun khỏc + Mc 0: Cha thờm hn hp dung dch chun vo 1000 àL mu th + Mc 1: Thờm 30àL hn hp dung dch chun ba cht nng 10 -3M vo 970àL mu th + Mc 2: Thờm 60àL hn hp dung dch chun ba cht nng 10 -3M vo 940 àL mu th + Mc 3: Thờm 90àL hn hp dung dch chun ba cht nng 10 -3M vo 910 àL mu th Mu th c pha nh sau: Mu bia chai H Ni c rung siờu õm, sau ú lc qua mng lc 0,45àm, Pha loóng 20 ln bng nc ion Mi mu th c o lp li ln Kt qu thu c trỡnh by bng 3.11, bng 3.12 v bng 3.13 Bng 3.11 Kt qu kho sỏt ỳng ca phng phỏp da trờn thờm chun oxalate -5 Nng oxalate chun thờm vo (1,00.10 M) Nng oxalate thu hi c (1,00.10-5M) Hiu sut thu hi (%) Hiu sut thu hi trung bỡnh (%) Mc 3,00 2,85 95,0 Mc 6,00 6,24 104 98,6 Mc 9,00 87,0 96,7 Bng 3.12 Kt qu kho sỏt ỳng ca phng phỏp da trờn thờm chun tartrate -5 Nng tartrate chun thờm vo (1,00.10 M) Nng tartrate thu hi c (1,00.10-5M) 42 Mc 3,00 2,91 Mc 6,00 5,94 Mc 9,00 9,09 Hiu sut thu hi (%) Hiu sut thu hi trung bỡnh (%) 97,0 99,0 99,0 101 Bng 3.13 Kt qu kho sỏt ỳng ca phng phỏp da trờn thờm chun citrate -5 Nng citrate chun thờm vo (1,00.10 M) Nng citrate thu hi c (1,00.10-5M) Hiu sut thu hi (%) Hiu sut thu hi trung bỡnh (%) Mc 3,00 2,85 95,0 Mc 6,00 6,36 106 99,7 Mc 9,00 8,82 98,0 Cỏc kt qu thu c bng 3.10 - 3.12 vi thu hi ca oxalate, tartrate v citrate khỏ cao, t 98,4ữ99,7%, cho thy phng phỏp cú ỳng cao T cỏc kt qu nghiờn cu thu c cho thy qui trỡnh phõn tớch ó ỏp ng c cỏc yờu cu v lp li v thu hi phõn tớch lng nh cỏc hp cht hu c nn mu phc lch chun tng i ca cỏc cht phõn tớch (RSD) nh hn 3, thu hi ca cỏc cht phõn tớch khong 98,6ữ99,7% Vi quy trỡnh phõn tớch ny chỳng tụi ỏp dng phng phỏp CE-C 4D phõn tớch mt s i tng mu thc t 3.3 Phõn tớch mu thc t 3.3.1 Phõn tớch mu bia Cỏc mu bia c la chn bao gm mt s loi bia chai v bia lon.Thụng tin v cỏc mu dc phm ny c nờu bng 3.14 Bng 3.14 Thụng tin v c im ca tng loi bia Tờn sn phm c im Xut x Bia chai H Ni Dng thnh phm: úng chai Tng cụng ty ru bia nc Dung tớch: 450ml gii khỏt H Nụi Dng thnh phm: úng lon Tng cụng ty ru bia nc Dung tớch: 330ml gii khỏt H Nụi Bia lon H Ni 43 Bia lon Halida Dng thnh phm: úng lon Nh mỏy Bia ụng Nam Dung tớch: 330ml Bia chai Saigon Dnh thnh phm: úng chai Tng cụng ty Ru bia Special Dung tớch: 330ml Nc gii khỏt Si Gũn Bia lon Tiger Dng thnh phm: úng lon Cụng ty TNHH nh mỏy Dung tớch: 330ml Bia Vit Nam Dng thnh phm: úng lon Cụng ty TNHH nh mỏy Dung tớch: 330ml Bia Vit Nam Bia lon Haliken Quy trỡnh x lý cỏc mu bia Rung siờu õm 30 phỳt (loi b khớ ga) Lc qua mng lc (0,45àm) Pha loóng vi t l thớch hp Mu bia chai H Ni c pha loóng 20 ln Mu bia lon H Ni c pha loóng ln Mu bia lon Halida c pha loóng 20 ln Mu bia chai Si Gũn Special c pha loóng 20 ln Mu bia lon Tiger c pha loóng 10 ln Mu bia lon Haliken pha loóng 10 ln Cỏc mu bia sau pha loóng c tin hnh phõn tớch iu kin ó ti 44 u Tin hnh phõn tớch oxalate, tartrate v citrate bng phng phỏp thờm chun Kt qu in di ca mu phõn tớch cha thờm chun c th hin hỡnh 3.11 n hỡnh 3.16 + Mu bia lon H Ni v mu bia chai H Ni 20mV Bia lon Hà Nôi Bia chai Hà Nội 100 200 300 400 500 Thời gian di chuyển (giây) 600 700 Hỡnh 3.11 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate mu bia chai H Ni v mu bia lon H Ni + Mu bia lon Halida v Mu bia chai Si Gũn Special 45 20mV Bia chai Sai Gòn Special Bia lon Halida 100 200 300 400 500 Thời gian di chuyển (giây) 600 700 Hỡnh 3.12 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate mu bia lon Halida v mu bia chai Si Gũn Special + Mu bia lon Haliken v Mu bia lon Tiger 50mV Bia lon tiger Bia lon Haliken 100 200 300 400 500 Thời gian di chuyển (giây) 600 700 Hỡnh 3.13 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate mu bia lon Haliken Cn c vo din tớch pic thu c trờn in di , bng phng phỏp thờm chun mt ln s xỏc nh c nng oxalate, tartrate v citrate cú mu bia lon Haliken Kt qu thu c nh bng 3.15 46 Bng 3.15 Kt qu xỏc nh hm lng cỏc cht oxalate, citrate mt s mu bia Mu Oxalate (mg/100ml) Citrate (mg/100ml) Bia lon H Ni - 9,40 0,1 Bia lon Halida - 23,0 2,0 Bia lon Tiger - 17,0 1,0 Bia lon Haliken - 12,0 0,7 Bia chai H Ni 2,10 0,1 11,6 0,8 Bia chai Si Gũn 1,57 0,08 10,4 0,6 Trong cỏc mu bia c la chn, khụng cú mu no xut hin tartrate Ch xỏc nh c oxalate mu hai mu bia chai (bia chai H Ni v bia chai Si Gũn) vi hm lng oxalate trung bỡnh hai mu l 1,84 mg/ml Citrate xut hin tt c cỏc mu bia Hm lng citrate cao nht mu bia lon Halida v thp nht mu bia lon H Ni Hm lng citrate trung bỡnh cỏc mu bia c la chn l 13,9 mg/ml 3.3.2 Phõn tớch mu mỡ gúi Quy trỡnh x lý mu Mu mỡ gúi c nghin nh, trn u, cõn mt lng nht nh trờn cõn phõn tớch (khong 0,5 g), thờm 4ml nc deion, rung siờu õm 30 phỳt, ly tõm (8000 rpm) 30 phỳt, lc qua mng lc 0,45àm, pha loóng vi t l thớch hp Mu mỡ gúi Ho Ho c pha loóng ln Mu mỡ gúi Ba Min pha loóng vi 20 Cỏc mu mỡ gúi sau pha loóng c phõn tớch bng thit b CE-C 4D Tin hnh phõn tớch oxalate, tartrate v citrate bng phng phỏp thờm chun in di phõn tớch mu m tụm Ba Min v mu m tụm Ho Ho cha thờm chun c th hin hỡnh 3.14 47 20mV Mỳ tôm Ba Miền Mỳ tôm Hảo Hảo 100 200 300 Thời gian di chuyển (giây) 400 500 Hỡnh 3.14 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate mu m tụm Ho Ho v mu m tụm Ba Min Cn c vo din tớch pic thu c trờn in di , bng phng phỏp thờm chun s xỏc nh c nng oxalate, tartrate v citrate cú mu mỡ gúi Ba Min Kt qu mu m tụm Ho Ho hm lng oxalate xỏc nh c l 22,0 2,0 mg/kg v hm lng citrate l 22,0 2,0 mg/kg Mu m tụm Ba Min hm lng oxalate xỏc nh c l 38,0 2,0mg/kg v hm lng citrate l 86,0 4,0 mg/kg Khụng phỏt hin tatrate cỏc mu m tụm c la chn 3.3.3 Phõn tớch mu tr tỳi lc lipton (tr trỏi cõy, v dõu) Quy trỡnh x lý mu Mu tr tỳi lc Lipton c xỏc nh lng trung bỡnh ca mt tỳi tr l 1,6gam La chn ngu nhiờn 01 tỳi tr, ngõm phỳt 150 ml nc deion ó un sụi (tng t quy trỡnh nh s dng) sau ú 45 phỳt nhit phũng Lc qua mng lc 0,45àm, pha loóng 20 ln v phõn tớch trờn thit b CE-C 4D Tin hnh phõn tớch oxalate, tartrate v citrate bng phng phỏp thờm chun Tin hnh phõn tớch in di vi cỏc iu kin ti u lp li ln ly kt qu 48 trung bỡnh xỏc nh hm lng cỏc cht oxalate, tartrate v citrate cú mu 3.3.4 Phõn tớch mu nc tr khụ va hố Quy trỡnh x lý mu Mu nc tr khụ va hố c lc qua mng lc 0,45àm sau ú pha loóng 20 ln trc thc hin phõn tớch trờn thit b CE-C 4D Tin hnh phõn tớch oxalate, tartrate v citrate bng phng phỏp thờm chun Tin hnh phõn tớch in di vi cỏc iu kin ti u lp li ln ly kt qu trung bỡnh xỏc nh hm lng cỏc cht oxalate, tartrate v citrate cú mu Kt qu in di ca mu phõn tớch v thờm chun c th hin hỡnh 3.15 20mV Trà khô vỉa hè Trà túi lọc Lipton 200 400 Thời gian di chuyển (giây) 600 Hỡnh 3.15 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate mu tr tỳi lc Lipton v mu tr khụ va hố Cn c vo din tớch pic thu c trờn in di , bng phng phỏp thờm chun s xỏc nh c nng oxalate, tartrate v citrate cú mu tr tỳi lc Lipton v mu nc tr khụ va hố Phng phỏp xỏc nh c hm lng oxalate mu tr tỳi lc Lipton 49 l 66,0 3,0 mg/kg, mu nc tr khụ va hố l 8,60 0,5 mg/kg v hm lng citrate mu tr tỳi lc Lipton l 0,94 0,05 mg/kg mu nc tr khụ va hố l 117 9,0 mg/kg Nh vy, s ba cht nghiờn cu, khụng phỏt hin tartrate bt c mu no, nhng phỏt hin citrate hoc ng thi oxalate v citrate mt s mu 3.3.5 Phõn tớch i chng phng phỏp CE-C4D vi phng phỏp HPLC kim chng kt qu phõn tớch bng phng phỏp CE-C 4D, chỳng tụi ó la chn 02 mu cú thnh phn phc nht l m tụm (nhón hiu Ho Ho v Ba Min) gi sang phõn tớch i chng ch tiờu quan trng l oxalate vi phng phỏp tiờu chun Sc ký lng hiu nng cao (HPLC) ti Vin Kim nghim an ton v sinh thc phm quc gia Cỏc kt qu ny c trỡnh by bng 3.15 Bng 3.16 So sỏnh kt qu phõn tớch i chng hm lng oxalate, citrate v tartrate mu m tụm Hm lng Oxalate (mg/kg) M tụm Ho Ho M tụm Ba Min % sai khỏc gia Hm lng cht phõn tớch Hm lng cht phõn thu c vi phng phỏp tớch thu c vi CE - C4D phng HPLC 22,0 2,0 25,16 12,6 38,0 2,0 42,01 9,5 hai phng phỏp T kt qu bng 3.16 cho thy, sai s gia hai phng phỏp di 15% iu ny chng t hm lng ca oxalate thu c theo hai phng phỏp khỏ phự hp Nh vy, cú th s dng phng phỏp CE-C 4D xỏc nh hm lng ca oxalate mu m tụm, bia, tr, v m rng nghiờn cu cho cỏc i tng mu khỏc 50 KT LUN Nh vy, quy trỡnh xỏc nh ng thi ba cht oxalate, tartrate v citrate mu thc phm bng phng phỏp in di mao qun s dng detector dn khụng tip xỳc ó c nghiờn cu phỏt trin vi nhng kt qu sau: ó ti u c cỏc iu kin tỏch ng thi ba cht oxalate, tartrate v citrate bng phng phỏp CE - C 4D Cỏc iu kin ti u bao gm: detector: CE C4D; dung dch din ly: m His 50mM MES 30mM (pH = 6,3), CTAB 25àM; Th tỏch: -15kV; S dng mao qun silica trn, tng chiu di 60 cm, chiu di hiu dng 50cm, ng kớnh 75àm; Thi gian bm mu: 30s; Phng phỏp bm mu: thy ng lc hc kiu xiphụng, h = 25cm Th t cỏc cht tỏch c vi mao qun silica l: oxalate, tartrate, citrate ó nghiờn cu nh hng ca mt s cht cú th xut hin cỏc nn mu nghiờn cu, ú chloride, formate cú nh hng ỏng k v xut bin phỏp khc phc ỏnh giỏ c phng phỏp phõn tớch: xõy dng ng chun xỏc nh cht khong nng 2,5 10 -4M, tớnh tuyn tớnh (cỏc giỏ tr R2 u ln hn 0,999), lp li (cỏc giỏ tr RSD nh hn 3), ỳng (hiu sut thu hi ca oxalate, tartrate v citratet khong 99% - 99,5%), gii hn phỏt hin (LOD) ca oxalate, tartrate l 2,5.10-6M v ca citrate l 5,0.10-6M; gii hn nh lng (LOQ) ca oxalate, tartrate l 8,3.10-6M v ca citrate l 16,7.10-6M Quy trỡnh ó c ỏp dng phõn tớch mt s mu thc phm (bia, mỡ gúi, tr) ó tin hnh phõn tớch i chng ch tiờu oxalate mu mỡ tụm vi phng phỏp tiờu chun HPLC Vin Kim nghim An ton thc phm thc hin cho kt qu ỏng tin cy Phng phỏp ny cho thy u im n gin, nhanh, d s dng, cú th hng ti mc tiờu ỏp dng ti cỏc phũng thớ nghim tuyn a phng nhm gúp phn kim tra mt s ch tiờu an ton thc phm, ỏp ng nhu cu thc t ca Vit 51 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dilutionmass spectrometry (ID-MS) and its usefulness in testing existing assays for urinary oxalate, Clinica Chimica Acta, 170:227-235 Corinne Rivasseau , Anne-Marie Boisson, Gaă elle Mong elard, Georgy Couram, Olivier Bastien, Richard Bligny (2006) Rapid analysis of organic acid in plant extracts by capillary electrophoresis with indirect UV detection Directed metabolic analyses during metal stress, Journal of Chromatography A, (112), 283290 52 Curhan (1999), GC epidemiologische Hinweise fỹr die Rolle des Oxalat- Nierensteine spontan, J Endourol 13 (9): 629-31 10 Hoppe B, Kemper MJ, Hvizd MG, Sailer DE, Langman CB (1998), Simultaneous determination of oxalate, citrate and sulfate in children's plasma with ion chromatography, Kidney Int, 53(5):1348-52 11 Mariỏn Masỏr et al (2003), Determination of oxalate in beer by zone electrophoresis on a chip with conductivity detection, Journal of seperation science, 26, 647-652 12 Michael Trevaskis, V Craige Trenerry (1996) An investigation into 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analysis of short-chain organic acid in coffee, Journal of Chromatography A, 1032: 299304 22 Xiumei Geng , Sufang Zhang , Qian Wang , Zongbao (Kent) Zhao (2008), Determination of organic acid in the presence of inorganic anions by ion chromatography with suppresse d conductivity detection, Journal of Chromatography A, 1192 : 187190 23 Fengwu Wu, Zhike He, Qingyao Luo, Yun'e Zeng (1999), HPLC determination of oxalic acid using tris(1,10-phenanthroline) ruthenium (II) chemiluminescence application to the analysis of spinach, Food Chemistry, (65),543-546 24 H.Z.Liana, L.Mao, X.L.Ye, J.Miao (1999), Simultaneous determination of oxalic, fumaric, maleic and succinic acid in tartaric and malic acid for pharmaceutical use by ion-suppression reversed-phase high performance liquid chromatography, Journal of Pharmaceutical and Biomedical Analysis, (19), 621-625 25 J G March, B M Simonet, F Grỏses, J A Muủoz, M Valiente (2003), Determination of trace amounts of oxalate in renal calculi and related samples by gas chromatography-mass spectrometry, Journal of Chromatography A,57(11), 811-817 26 Thanh Duc Mai, Peter C Hauser (2012), Simultaneous separations of cations and anions by capillary electrophoresis with contactless conductivity detection employing a sequential injection analysis manifold for flexible manipulation of sample plugs, Journal of Chromatography A, 1267, 266 272 27 Thanh Duc Mai, Peter C Hauser (2011), Anion separations with pressure54 assisted capillary electrophoresis using a sequential injection analysis manifold and contactless conductivity detection, Electrophoresis, 32, 3000 3007 28 Virginia Garca-Canas, Miguel Herrero, Carolina Simo, Alejandro Cifuentes (2010), Recent advances in the application of capillary electromigration methods for food analysis and Foodomics, Electrophoresis, (31), 20522 55 [...]... dung dch cht 13 in ly cú cht m pH v trong tỏc dng ca in trng nht nh do th (V) t vo hai u mao qun sinh ra Cu to ca h in di c cho trong hỡnh 1.1 [1] Hỡnh 1.1 S phõn tớch h in di mao qun in di, tc in di v thi gian in di Trong in di, tc di chuyn ca cỏc ht tớch in (v) t l thun vi cng in trng (E): v = E Trong ú: (1.1) v: tc di chuyn ca ion : l din di E: l in trng ngoi Trong iu kin tc ca dũng n nh, lc... phõn tớch h in di mao qun 14 in di, tc in di v thi gian in di .14 1.6.1 .Mao qun 15 Hỡnh 1.2 Mt ct ngang ca mao qun 15 Hỡnh 1.3 Lp in tớch kộp trờn b mt mao qun 16 Hỡnh 1.4 nh hng ca dũng EOF n tc ca cỏc ion trong quỏ trỡnh in di .16 1.6.2.Dung dch m pH v pha ng trong phng phỏp in di mao qun .17 Bng 1.2 Cỏc cht thng dựng lm pha ng trong CE v giỏ... quan trng trong phng phỏp in di mao qun õy chớnh l mt trong cỏc yu t quyt nh s in di hn hp ca cỏc cht mu Mao qun c ch to ch yu l silica c gi l mao qun silica Trong mt s trng hp cng cú th dựng mao qun teflon, khi mao qun silica khụng phự hp, vớ d mu cú ion F- v tỏch pH thp [1] Hỡnh 1.2 Mt ct ngang ca mao qun Lp in kộp trờn thnh mao qun v dũng in di thm thu Trong quỏ trỡnh in di, lp in kộp v th ca chỳng... 7,10; 13,30 DIPSO 7,50 Histidine 1,78; 5,97; 8,97 HEPFS 7,51 Lysine 2,20; 8,90; 10,28 HEPPSO 7,9 Arginine 2,18; 9,09; 13,20 1.6.3 Ngun in th cao Quỏ trỡnh in di trong mao qun ch xy ra khi cú ngun th V mt chiu nht nh t vo hai u mao qun Th V ny to ra lc in trng E v dũng in I trong mao qun, nú iu khin v duy trỡ s in di ca cỏc cht Trong in di mao qun, in th V mt chiu thng c dựng t vo hai u mao qun l t... l t 150 550 V/1cm mao qun Tuy 17 nhiờn, th V c dựng phi lm sao khụng cho dũng in I quỏ ln trong mao qun, dũng in I ny ch nờn nm trong vựng t 10 75 àA Vic chn in th V l bao nhiờu tựy thuc vo bn cht ca cht phõn tớch, cht nn ca mu, giỏ tr pH ca pha ng in di, [1] 1.6.4 K thut bm mu trong phng phỏp in di mao qun Trong in di mao qun cú ba phng phỏp thng dựng np mu phõn tớch vo trong mao qun, bao gm: Phng... vụ c vi hiu xut thu hi trong khong 88% - 105%, RSD < 7% Phng phỏp cú th ỏp dng trong cỏc nn mu phc tp, trong ú cú mu hn hp lờn men 1.5.4 Cỏc phng phỏp in di mao qun in di mao qun l mt k thut tỏch cht phõn tớch l cỏc ion hoc cỏc cht khụng ion nhng cú mi liờn h cht ch vi cỏc ion trong mt mao qun hp cha y dung dch m, t trong in trng; do linh ng in di ca cỏc ion khỏc nhau, chỳng di chuyn vi tc khỏc nhau... ca pH n din tớch pic (Spic) v thi gian di chuyn (tdc) ca oxalate, tartrate v citrate 30 Hỡnh 3.5 in di kho sỏt chiu cao bm mu xỏc nh iu kin ti u xỏc nh ng thi 3 cht1-oxalate 5,0.10-4 M, 2-citrate 5,0.10-4M v 3-tartrate 5,0.10-4M Mao qun silica ng kớnh trong 75àm, chiu di 60cm, chiu di hiu dng 50cm, th tỏch -15kV, thi gian bm mu 30s 33 Hỡnh 3.6 in di kho sỏt th in di xỏc... 45 Hỡnh 3.12 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate trong mu bia lon Halida v mu bia chai Si Gũn Special 46 Hỡnh 3.13 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate trong mu bia lon Haliken .46 Bng 3.15 Kt qu xỏc nh hm lng cỏc cht oxalate, citrate trong mt s mu bia .47 Hỡnh 3.14 in di phõn tớch 1-oxalate, 2-tartrate, 3-citrate trong mu m tụm Ho Ho v mu m tụm... mu trong phng phỏp in di mao qun 18 Hỡnh 1.5 Cỏc k thut bm mu trong phng phỏp in di mao qun 18 1.7.Phng phỏp in di mao qun s dng detector o dn khụng tip xỳc (CE - C4D) 19 Hỡnh 1.6 Nguyờn lý hot ng ca cm bin o dn khụng tip xỳc 20 Hỡnh 1.7 S biu din cu trỳc (A) v mch in tng ng (B) ca cm bin o dn khụng tip xỳc 20 CHNG 2: THC NGHIM 21 Hỡnh 2.1 H thit b in di. .. silica, chiu di hiu dng 30cm, o ngc dũng in di thm thu v th in di l -30 kV Dung dch m cha 3.10-4 mol/L tetradecyltrimethylammonium v 10- 2 mol/L trimellitate pH = 9,0 Quỏ trỡnh tỏch t c trong 100s Khong tuyn tớnh i vi oxalate t 6.10- 5 n 10- 3 mol/L 1.6 Phng phỏp in di mao qun in di mao qun l mt k thut tỏch cỏc cht da trờn s di chuyn khỏc nhau ca cỏc phõn t cht (ch yu l cỏc ion mang in tớch) trong dung