氨基酸的多元校正滴定法测定
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摘要
氨基酸的测定在理论上和实际应用中都有重要的意义。目前氨基酸的分析方法主要有化学分析法、光谱分析法、电化学分析法和色谱分析法等,但利用多元校正法对氨基酸进行测定的研究尚不多见。本文以氢离子选择性电极为工作电极,根据氨基酸的酸性或碱性,以一定浓度的盐酸或氢氧化钠为滴定剂,采用多元校正滴定法,对具有代表性的氨基酸的单组分溶液以及它们的双组分混合溶液分别进行测定。本文主要做的工作如下:(1)从四种酸碱滴定形式(包括酸直接滴定、碱直接滴定、酸返滴定、碱返滴定)中,分别确定了测定各类(包括酸性、碱性和中性)单组分氨基酸及它们的双组分混合氨基酸的最佳滴定形式。(2)采用最佳滴定形式,对谷氨酸、丝氨酸、精氨酸和组氨酸的单组分以及它们的各双组分混合氨基酸进行了测定,其中单组分测定结果误差控制在1%左右,而双组分测定结果误差控制在3%左右。(3)针对四种滴定形式,对各种类型的单组分氨基酸和双组分混合氨基酸滴定曲线的形成原因,进行了较系统的讨论和解释。
The determination of amino acid has significant meaning in both theory field and application field. Nowadays there are various methods of amino acid determination, including chemical analysis, spectrum analysis, electrical analysis and chromatography analysis. However, it is rare to use multivariate calibration to measure amino acid. In this thesis, the determination of single component amino acid and double components mixed amino acid is carried out through multivariate calibration titration with solution of NaOH or HC1 as the titrant, hydrogen ion selective electrode as the indicator electrode, while automatic titrator is operate instrument. Hereby, following jobs have been done: (1) The best titration way for determination of each single acidic, alkalescent, neutral amino acid component and its various double mixed components is summarized through reviewing the four titration ways, including direct titration with acid, direct titration with alkali, back titration with acid and back titration with alkali. (2) Through the best way of titration, desirable determination outcome of Glu, His, Arg and Ser single component and its mixed double components is obtained, the errors is about 1% for single component while about 3%for double components. (3) Each titration curve of single amino acid component and double mixed components, obtained from every titration way, is systematically discussed and explained.
The determination of amino acid has significant meaning in both theory field and application field. Nowadays there are various methods of amino acid determination, including chemical analysis, spectrum analysis, electrical analysis and chromatography analysis. However, it is rare to use multivariate calibration to measure amino acid. In this thesis, the determination of single component amino acid and double components mixed amino acid is carried out through multivariate calibration titration with solution of NaOH or HC1 as the titrant, hydrogen ion selective electrode as the indicator electrode, while automatic titrator is operate instrument. Hereby, following jobs have been done: (1) The best titration way for determination of each single acidic, alkalescent, neutral amino acid component and its various double mixed components is summarized through reviewing the four titration ways, including direct titration with acid, direct titration with alkali, back titration with acid and back titration with alkali. (2) Through the best way of titration, desirable determination outcome of Glu, His, Arg and Ser single component and its mixed double components is obtained, the errors is about 1% for single component while about 3%for double components. (3) Each titration curve of single amino acid component and double mixed components, obtained from every titration way, is systematically discussed and explained.
引文
铩颷1]P.W库彻,G.B罗尔斯顿.生物化学.北京:科学出版社,2002
[2]聂剑初.生物化学简明教程.北京:高等教育出版社,1999
[3]分析化学手册.第一分册.第二版.北京:化学工业出版社,1997
[4]李元宗,常文保.生化分析.高等教育出版社,2003
[5]Guozhen Fang,Nie Liu.Determination of eight essential amino acids in mixtures bychemometrics-spectrophotometry without separation.Analytica Chimica Acta.2001,445:245-253
[6]Dimas A.M.Zaia,Kelly C.L.Ribas,Ca'Ssia T.B.V.Zaia.Spectrophotometric determination ofcysteine and/or carbocysteine in a mixture of amino acids,shampoo,and pharmaceuticalproducts using p-benzoquinone.Talanta.1999,50:1003-1010
[7]刘效兰.氨基酸荧光分析法应用.理化检验-化学分册.2004,40(8):457-458
[8]陈悦娇,马应丹,胡翔蕤.荧光光度法测定白果中游离氨基酸含量.广州食品工业科技.2003,19(2):87-89
[9]刘彩虹,李红.氨基酸的电分析进展.氨基酸和生物资源.2003,25(4):71-74
[10]邹永德,王进.用碳糊修饰电极测定氨基酸的伏安法研究.分析测试学报.1999,18(2):25-28
[11]王长发,李健全.赖氨酸与邻氯苯甲醛的衍生化反应及其示波极谱法测定.分析化学.1992,20(4):433-436
[12]蔡凌霜,徐昊,张玉清等.非水滴定测定α氨基酸含量实验的微型化.分析科学学报.2003,19(6):570-572
[13]陈志慧.氨基酸分析技术的综述.广东化工.2004,2:16-20
[14]Erik Gerlo,Willy Lissens,Gerda Winckelmans,et al.Gas chromatographic-massspectrometric analysis of N-acetylated amino acids:The first case of aminoacylase Ideficiency.Analytica Chimica Acta.2006,571:191-199
[15]常碧影,杨本军.氨基酸分析技术的综述.现代科学仪器.2004,6:7-10
[16]J.L.G'Omez-Ariza,M.J.V.-E,V.Bemal-Daza.Characterization and analysis of amino acidsin orange juice by HPLC-MS/MS for authenticity assessment.Analytica Chimica Acta.2005,540:221-230
[17]Liu-Yin Fan,Hong-Li Chen,Ji-You Zhang,et al.Continuous on-line derivatization anddetermination of amino acids by a microfluidic capillary eleetrophoresis system with acontinuous sample introduction interface.Analytica Chimica Acta.2004,501:129-135
[18]Juan Antonio Calzado,Cristina Palet,Manuel Valiente.Facilitated transport and separation ofaromatic amino acids through activated composite membranes.Analytica Chimica Acta.2001,431:59-67
[19]Hiroki Hosono,Junji Fukuda,Hiroaki Suzuki.,et al.On-chip handling of solutions and electrochemiluminescence detection of amino acids.Sensors and Actuators.2007,122:
542-548
铩颷20]于泓,牟世芬.氨基酸分析方法的研究进展.分析化学.2005,33(3):394-400
[21]戴红.氨基酸分析的检测方法评述.皮革科学与工程.2004,14(3):39-43
[22]Ming-Mu Hsieh,Sheng-Min Chen.Determination of amino acids in tea leaves and beveragesusing capillary electrophoresis with light-emitting diode-induced fluorescence detectio.Talanta.2007,981:51-53
[23]Min Yang,Sterling A,Tomellini.An HPLC detection scheme for underivatized amino acidsbased on tryptophan fluorescence recovery.Analytica Chimica Acta.1999,409:45-53
[24]Lingberg W,Kowalski B.Evaluation of potentiomectric acid-basic titrations partial leastsquares calibration.Analytica Chimica Acta.1988,206:125-135
[25]张大伦.多元线性回归同时单点pH滴定法研究.分析科学学报.1996,12(4):294-297
[26]朱仲良,李通化,鲁准军.两维pH滴定法同时测定混合极弱酸.高等学校化学学报.1995,16(9):1368-1370
[27]Bela Noszal,Maria Juhasz.Determination of sulphate by pH titration.Talanta.1987,34:397-400
[28]Issam Jano,James Hardcastle,Lamia A.Jano,et al.General equation for determining thedissociation constants of polyprotic acids and bases from additive properties:Part Ⅳ.Application to potentiometric titration.Analytica Chimica Acta.2001,428:309-321
[29]倪永年,金玲.计算滴定分析法应用及进展.分析化学.1996,24(10):1219-1226
[30]Yongnian Ni.Simultaneous determination of mixtures of acids by potentiometric titration.Analytica Chimica Acta.1998,367:145-152
[31]朱仲良,褚建新,李通化等.两维酸碱-沉淀滴定法同时测定有机酸.分析化学.1996,24(6):643-648
[32]朱仲良,赵国华,李通化等.pH滴定-主成分回归法同时测定酚类同系物分析化学.分析化学.1996,24(1):87-89
[33]Yongnian Ni,Zhaohong Peng.Determination f mixed metal ions by complex metric titrationand nonlinear partial least squares calibration.Analytica Chimica Acta.1995,304:217-222
[34]乔沿江,王英华,刘友谊.配位滴定法同时测定金属离子浓度的研究.长春地质学院学报.1994,24(3):339-314,348
[35]张云,孙健,于学涛等.非平衡态-恒电位配位滴定法同时测定铝和铁.分析化学.2005,33(4):1764-1766
[36]张云,刘婷婷,孙健.氟-铁恒电位配位滴定法测定铁离子的研究.分析科学学报.2006,22(4):451-453
[37]张云,褚亮亮,许圆.恒电位氟-铝配位滴定法测定铝的研究.分析实验室.2005,24(3):44-47
[38]倪永年,金玲,吴英亮.主成分-络合滴定法测定混合金属离子.高等学校化学学报.1998,19(3):359-362
[39]张大伦.多元线性回归pH配位滴定法同时测定镍铅钴.武汉化工学院学报.1996, 18(2):1-4
铩颷40]张云,李大鹏,杨立等.氟-铝络合滴定计算分析法测定乙醇-水溶液介质中的铝.分析化学.2005,33(7):947-950
[41]Yongnian Ni,Yingliang Wu.Simultaneous determination of mixture of metal ions bycomLexometric titration and multivariate calibration.Analytica Chimica Acta.1997,354:233-240
[42]童庆,李晔,樊霆等.多元线性回归pH滴定同时测定垃圾渗滤液中重金属浓度.环境监测管理与技术.2005,17(6):27-29
[43]刘婷婷.恒电位法对氨基酸的测定[硕士学位论文].上海:同济大学化学系,2007
[44]张云.分析化学.同济大学出版社,2003
[2]聂剑初.生物化学简明教程.北京:高等教育出版社,1999
[3]分析化学手册.第一分册.第二版.北京:化学工业出版社,1997
[4]李元宗,常文保.生化分析.高等教育出版社,2003
[5]Guozhen Fang,Nie Liu.Determination of eight essential amino acids in mixtures bychemometrics-spectrophotometry without separation.Analytica Chimica Acta.2001,445:245-253
[6]Dimas A.M.Zaia,Kelly C.L.Ribas,Ca'Ssia T.B.V.Zaia.Spectrophotometric determination ofcysteine and/or carbocysteine in a mixture of amino acids,shampoo,and pharmaceuticalproducts using p-benzoquinone.Talanta.1999,50:1003-1010
[7]刘效兰.氨基酸荧光分析法应用.理化检验-化学分册.2004,40(8):457-458
[8]陈悦娇,马应丹,胡翔蕤.荧光光度法测定白果中游离氨基酸含量.广州食品工业科技.2003,19(2):87-89
[9]刘彩虹,李红.氨基酸的电分析进展.氨基酸和生物资源.2003,25(4):71-74
[10]邹永德,王进.用碳糊修饰电极测定氨基酸的伏安法研究.分析测试学报.1999,18(2):25-28
[11]王长发,李健全.赖氨酸与邻氯苯甲醛的衍生化反应及其示波极谱法测定.分析化学.1992,20(4):433-436
[12]蔡凌霜,徐昊,张玉清等.非水滴定测定α氨基酸含量实验的微型化.分析科学学报.2003,19(6):570-572
[13]陈志慧.氨基酸分析技术的综述.广东化工.2004,2:16-20
[14]Erik Gerlo,Willy Lissens,Gerda Winckelmans,et al.Gas chromatographic-massspectrometric analysis of N-acetylated amino acids:The first case of aminoacylase Ideficiency.Analytica Chimica Acta.2006,571:191-199
[15]常碧影,杨本军.氨基酸分析技术的综述.现代科学仪器.2004,6:7-10
[16]J.L.G'Omez-Ariza,M.J.V.-E,V.Bemal-Daza.Characterization and analysis of amino acidsin orange juice by HPLC-MS/MS for authenticity assessment.Analytica Chimica Acta.2005,540:221-230
[17]Liu-Yin Fan,Hong-Li Chen,Ji-You Zhang,et al.Continuous on-line derivatization anddetermination of amino acids by a microfluidic capillary eleetrophoresis system with acontinuous sample introduction interface.Analytica Chimica Acta.2004,501:129-135
[18]Juan Antonio Calzado,Cristina Palet,Manuel Valiente.Facilitated transport and separation ofaromatic amino acids through activated composite membranes.Analytica Chimica Acta.2001,431:59-67
[19]Hiroki Hosono,Junji Fukuda,Hiroaki Suzuki.,et al.On-chip handling of solutions and electrochemiluminescence detection of amino acids.Sensors and Actuators.2007,122:
542-548
铩颷20]于泓,牟世芬.氨基酸分析方法的研究进展.分析化学.2005,33(3):394-400
[21]戴红.氨基酸分析的检测方法评述.皮革科学与工程.2004,14(3):39-43
[22]Ming-Mu Hsieh,Sheng-Min Chen.Determination of amino acids in tea leaves and beveragesusing capillary electrophoresis with light-emitting diode-induced fluorescence detectio.Talanta.2007,981:51-53
[23]Min Yang,Sterling A,Tomellini.An HPLC detection scheme for underivatized amino acidsbased on tryptophan fluorescence recovery.Analytica Chimica Acta.1999,409:45-53
[24]Lingberg W,Kowalski B.Evaluation of potentiomectric acid-basic titrations partial leastsquares calibration.Analytica Chimica Acta.1988,206:125-135
[25]张大伦.多元线性回归同时单点pH滴定法研究.分析科学学报.1996,12(4):294-297
[26]朱仲良,李通化,鲁准军.两维pH滴定法同时测定混合极弱酸.高等学校化学学报.1995,16(9):1368-1370
[27]Bela Noszal,Maria Juhasz.Determination of sulphate by pH titration.Talanta.1987,34:397-400
[28]Issam Jano,James Hardcastle,Lamia A.Jano,et al.General equation for determining thedissociation constants of polyprotic acids and bases from additive properties:Part Ⅳ.Application to potentiometric titration.Analytica Chimica Acta.2001,428:309-321
[29]倪永年,金玲.计算滴定分析法应用及进展.分析化学.1996,24(10):1219-1226
[30]Yongnian Ni.Simultaneous determination of mixtures of acids by potentiometric titration.Analytica Chimica Acta.1998,367:145-152
[31]朱仲良,褚建新,李通化等.两维酸碱-沉淀滴定法同时测定有机酸.分析化学.1996,24(6):643-648
[32]朱仲良,赵国华,李通化等.pH滴定-主成分回归法同时测定酚类同系物分析化学.分析化学.1996,24(1):87-89
[33]Yongnian Ni,Zhaohong Peng.Determination f mixed metal ions by complex metric titrationand nonlinear partial least squares calibration.Analytica Chimica Acta.1995,304:217-222
[34]乔沿江,王英华,刘友谊.配位滴定法同时测定金属离子浓度的研究.长春地质学院学报.1994,24(3):339-314,348
[35]张云,孙健,于学涛等.非平衡态-恒电位配位滴定法同时测定铝和铁.分析化学.2005,33(4):1764-1766
[36]张云,刘婷婷,孙健.氟-铁恒电位配位滴定法测定铁离子的研究.分析科学学报.2006,22(4):451-453
[37]张云,褚亮亮,许圆.恒电位氟-铝配位滴定法测定铝的研究.分析实验室.2005,24(3):44-47
[38]倪永年,金玲,吴英亮.主成分-络合滴定法测定混合金属离子.高等学校化学学报.1998,19(3):359-362
[39]张大伦.多元线性回归pH配位滴定法同时测定镍铅钴.武汉化工学院学报.1996, 18(2):1-4
铩颷40]张云,李大鹏,杨立等.氟-铝络合滴定计算分析法测定乙醇-水溶液介质中的铝.分析化学.2005,33(7):947-950
[41]Yongnian Ni,Yingliang Wu.Simultaneous determination of mixture of metal ions bycomLexometric titration and multivariate calibration.Analytica Chimica Acta.1997,354:233-240
[42]童庆,李晔,樊霆等.多元线性回归pH滴定同时测定垃圾渗滤液中重金属浓度.环境监测管理与技术.2005,17(6):27-29
[43]刘婷婷.恒电位法对氨基酸的测定[硕士学位论文].上海:同济大学化学系,2007
[44]张云.分析化学.同济大学出版社,2003