草棉花活性组黄酮醇类多成分质谱定量及其体内代谢产物分析方法比较
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摘要
本文对来源于新疆特有具有抗老年痴呆和抗血栓活性的民族药用植物草棉活性提取物AB-8-2中异槲皮苷(isoquercetrin)、槲皮黄苷(quercimeritrin)、槲皮素-3'-葡萄糖苷(quercetin-3'-glucoside)、8-甲氧基-山奈酚-7-鼠李糖苷(8-methoxyl-kaempferol-7-rhamnoside)、紫云英苷(astragalin)、山奈酚-3-(6"-对-香豆酰)-葡萄糖苷(kaempferol-3-(6"-P-coumaroyl)-glucoside)、芦丁(rutin)、槲皮素(quercetin)、山奈酚(kaempferol)等采用HPLC-MS~n技术,开展以多反应监测(MRM)检测模式的9组分同时定量研究。同时运用该方法成功的对3个不同批次的AB-8-2样品进行了9组分定量,显示了良好的重现性和准确度。
MS~E或IDA是通过合理地设置质谱的扫描方式可以迅速获得生物样品中被分析测试物的多维信息的LC-MS/MS技术。根据大多数药物的代谢产物基本保持了原药的骨架特征抑或是亚骨架特征,代谢物的质谱裂解行为与原药相近,综合运用LC-MS/MS技术可迅速找到可能的代谢物,并进行结构鉴定的这一理论,初步探讨了大鼠灌胃该活性提取物后的尿液样品与胆汁样品中的代谢产物进行IDA与MS~E代谢产物分析,并对其代谢产物进行了这两种代谢产物分析方法的方法学比较研究。为整体研究中药的代谢及其辩证的作用机制作基础性研究,另外,通过分析这种高通量式的代谢产物信息获取及分析技术,为研究疾病本身提供理论依据。
For the modernization of traditional Chinese herbal medicine, quanlity control and the establishment of dependenble pharmacological assessment system for the crude materials and products are considered as the two principal challenges. And at present, it is not enough to evaluate the quality of TCM using one single component. Simultaneous quantitation of multi-components on the basis of pharmacological study has attracted the interests of researchers in the quanlity control of the TCM.
AB-8-2 was an active fraction obtained from Gossypium herbaceam L, which mainly consisted of flavonol glycosides. In this study, a method using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for the simultaneously determination of the 9 major constituents of this active fraction. And this method was successfully applied to the quantification of nine constituents in 3 AB-8-2 samples. Data acquisition under MS/MS was achieved by applying multiple reaction monitoring (MRM) of two fragment ions transitions to provide a high degree of sensitivity and specificity for both quantification and confirmation. Nine constituents are rutin, isoquercetrin, quercimeritrin, quercetin-3'-glucoside, 8-methoxyl-kaempferol-7-rhamnoside, astragalin, kaempferol-3-(6"-P-coumaroyl)-glucoside, quercetin, kaempferol eta.
The metabolisms in bile and urine could reflect the whole profiles of biotransformation of the parent medicine or compounds. In this paper, the metabolites of flavonols in the bile and urine samples after oral administistration of AB-8-2 were analyzed using LC-MS/MS in a single chromatographic run. Data were aquried using RRLC~(TM) coupled with a QTOF mass spectrometer. The Information Depended Acquirer (IDA) and MS~E (where E represents collision energy) data acquisition methods are compared and it was shown that the latter has been provide to be a powerful approach for the rapid metabolites identification.
Based on the IDA method established, the urine sample after oral administration of AB-8-2 was analyzed. In one analytical run, 38 constituents including 26 glucuronidated metabolites, 5 sulfation metabolites, 4 methylation metabolites , 3 flavonol aglycones and 1 glucuronidated conjudge methylation metabolite were characterized. Flavonol diglycosides in AB-8-2 probably remained one glycoside in the structure during the absorption process in intestinal tract. In bile sample, 3 constituents including 1 glucuronidated metabolite, 2 sulfation metabolites were characterized.
Based on the MS~E method established, the urine sample after oral administration of AB-8-2 was analyzed. In one analytical run, 34 constituents including 25 glucuronidated metabolites, 4 sulfation metabolites, 3 methylation metabolites , 4 flavonol aglycones and 1 glucuronidated conjudge sulfation metabolite were characterized. In bile sample, 15 constituents including 13 glucuronidated metabolites, 1 flavonol aglycones metabolites and 1 methylation metabolite were characterized.
In comparision with MS~E, IDA is an artificial intelligence-based produce ion scan mode providing automatic“on-the-fly”MS to MS/MS switching. And the fragmentation spectrometry was more characteristic. However, MS~E produced the same product ion profile but more comprehensive and overall. Although since no precursor-ion selection occurs in the mass spectrometer good chromatographic separation is a critical factor in affording fragment ions that are derived predominantly from the analyte of interest, MS~E enables the almost simultaneous acquisition of both LC-MS and fragmentation data from a single experiment. Conversely, IDA has its inherent drawback that has the capacity to generate some non-compound-related MS/MS spectra or miss potential metabolites.
MS~E或IDA是通过合理地设置质谱的扫描方式可以迅速获得生物样品中被分析测试物的多维信息的LC-MS/MS技术。根据大多数药物的代谢产物基本保持了原药的骨架特征抑或是亚骨架特征,代谢物的质谱裂解行为与原药相近,综合运用LC-MS/MS技术可迅速找到可能的代谢物,并进行结构鉴定的这一理论,初步探讨了大鼠灌胃该活性提取物后的尿液样品与胆汁样品中的代谢产物进行IDA与MS~E代谢产物分析,并对其代谢产物进行了这两种代谢产物分析方法的方法学比较研究。为整体研究中药的代谢及其辩证的作用机制作基础性研究,另外,通过分析这种高通量式的代谢产物信息获取及分析技术,为研究疾病本身提供理论依据。
For the modernization of traditional Chinese herbal medicine, quanlity control and the establishment of dependenble pharmacological assessment system for the crude materials and products are considered as the two principal challenges. And at present, it is not enough to evaluate the quality of TCM using one single component. Simultaneous quantitation of multi-components on the basis of pharmacological study has attracted the interests of researchers in the quanlity control of the TCM.
AB-8-2 was an active fraction obtained from Gossypium herbaceam L, which mainly consisted of flavonol glycosides. In this study, a method using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for the simultaneously determination of the 9 major constituents of this active fraction. And this method was successfully applied to the quantification of nine constituents in 3 AB-8-2 samples. Data acquisition under MS/MS was achieved by applying multiple reaction monitoring (MRM) of two fragment ions transitions to provide a high degree of sensitivity and specificity for both quantification and confirmation. Nine constituents are rutin, isoquercetrin, quercimeritrin, quercetin-3'-glucoside, 8-methoxyl-kaempferol-7-rhamnoside, astragalin, kaempferol-3-(6"-P-coumaroyl)-glucoside, quercetin, kaempferol eta.
The metabolisms in bile and urine could reflect the whole profiles of biotransformation of the parent medicine or compounds. In this paper, the metabolites of flavonols in the bile and urine samples after oral administistration of AB-8-2 were analyzed using LC-MS/MS in a single chromatographic run. Data were aquried using RRLC~(TM) coupled with a QTOF mass spectrometer. The Information Depended Acquirer (IDA) and MS~E (where E represents collision energy) data acquisition methods are compared and it was shown that the latter has been provide to be a powerful approach for the rapid metabolites identification.
Based on the IDA method established, the urine sample after oral administration of AB-8-2 was analyzed. In one analytical run, 38 constituents including 26 glucuronidated metabolites, 5 sulfation metabolites, 4 methylation metabolites , 3 flavonol aglycones and 1 glucuronidated conjudge methylation metabolite were characterized. Flavonol diglycosides in AB-8-2 probably remained one glycoside in the structure during the absorption process in intestinal tract. In bile sample, 3 constituents including 1 glucuronidated metabolite, 2 sulfation metabolites were characterized.
Based on the MS~E method established, the urine sample after oral administration of AB-8-2 was analyzed. In one analytical run, 34 constituents including 25 glucuronidated metabolites, 4 sulfation metabolites, 3 methylation metabolites , 4 flavonol aglycones and 1 glucuronidated conjudge sulfation metabolite were characterized. In bile sample, 15 constituents including 13 glucuronidated metabolites, 1 flavonol aglycones metabolites and 1 methylation metabolite were characterized.
In comparision with MS~E, IDA is an artificial intelligence-based produce ion scan mode providing automatic“on-the-fly”MS to MS/MS switching. And the fragmentation spectrometry was more characteristic. However, MS~E produced the same product ion profile but more comprehensive and overall. Although since no precursor-ion selection occurs in the mass spectrometer good chromatographic separation is a critical factor in affording fragment ions that are derived predominantly from the analyte of interest, MS~E enables the almost simultaneous acquisition of both LC-MS and fragmentation data from a single experiment. Conversely, IDA has its inherent drawback that has the capacity to generate some non-compound-related MS/MS spectra or miss potential metabolites.
引文
[1]孙汉董.现代化是中药与植物药发展的必由之路.云南中医学院学报. 2004; 27: 3-5
[2]蒋文跃.寻找物质基础非当前中药现代化关键.中草药.2003;34:1-3.
[3]李忠红杜冠华.中药质量监控之我见.世界科学技术-中医药现代化
质量标准研究. 2004;6: 67-71.
[4]耿平博士毕业论文,中国医学科学院协和医科大学. 2007
[5]高莉斯拉甫·艾白,新疆维药与国内外重要质量标准研究的比较.中国民族医药杂志. 2006; 4:29-30.
[6]李斌博士毕业论文,中国医学科学院协和医科大学. 2005
[7]彭涛雍炜安娟等,反相高效液相色谱/质谱法同时测定肉鸡中5种喹诺酮药物残留. Chinese Journal of Analytical Chemistry.2006; 34: 10-14.
[8] Pierluigi Caboni, Giorgia Sarais,Marco Cabras, and Alberto angioni. Determination of 4-Ethylphenol and 4-Ethylguaiacol in Wines by LC-MS-MS and HPLC-DAD-Fluorescence. J.Agric.Food Chem. 2007; 55:7288-7293
[9] Jacek Bielawski, Zdzislaw M. Szulc, Yusuf A. Hannun, Alicja Bielawska. Simultaneous quantitative analysis of bioactive sphingolipids by high-performance liquid chromatography-tandem mass spectrometry. Methods.2006; 39: 82-91.
[10] Gibson GG, Skett P. Pathways of Drug Metabolism. In Introduction to Drug Metabolism, Gibson GG, Skett P(ed.). Blackie Academic and Professional:London,1994;1-34
[11] Mulder GJ, Coughtrie MWH, Burchell B. Glucuronidation. In Conjugation Reactions in Drug Metabolism: an Integrated Approach.Mulder GJ(ed).taylor and Francis: London,1990; 42-91.
[12]陈义,高通微量分析.化学进展Progress in Chemistry. 2005; 17: 573-580.
[13] Charanjit sandhu, Johannes A. Hewel, Gwenael badis, Shaheynoor Taluder, Jian Liu, Timothy R.Hughes, and Andrew Emili. Evaluation of Data-dependent versusTargeted Shotgun Proteomic Approaches for Monitoring Transcription Factor Expression in Breast Cancer.Journal of proteome research. 2008;7:1529-1541.
[14] Geng P, Zhang RP, Aisa HA, He JM, Qu K, Zhu HB, Abliz Z. Fast profiling of the integral metabolism of flavonols in the active fraction of Gossypium herbaceam L. using liquid chromatography/multi-stage tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2007;21: 1877-88
[15]高洁王素娟徐瑞明杨永春张守仁司伊康石建功.以体内吸收分布特点指导花锚成分的研究.药学学报. 2004; 39: 198-203.
[16] Asish B.Chakraborty, Scott J.Berger and John C.Gebler. Use of an integrated MS-multiplexed MS/MS data acquisition strategy for high-coverage peptide mapping studies. Rapid Commun. Mass Spectrom. 2007;21:730-744.
[17] Keyume Ablajan, Zeper Abliz, Xiao-ya Shang, Jiu-ming He, Rui-ping Zhang and Jian-Gong Shi. Structural characterization of flavonol 3,7-di-O-glycosides and determination of the glycosylation position by using negative ion electrospray ionization tandem mass spectrometry. J.Mass Spectrum. 2006; 41:352-360.
[18] P.Kachlicki, J.Einhorn, D.Muth, L.Kerhoas and M.Stobiecki. Evaluation of glycosylation and malonylation patterns in flavonoid glycosides during LC/MS/MS metabolite profiling. J.Mass Spectrum. 2008;43:572-586.
[19] Lucia Politi, Luca Morini and Aldo Polettini. A driect screening procedure for diuretics in human urine by liquid chromatography-tandem mass spectrometry with information depengent acquisition. Clin Chim Acta. 2007;386(1-2):46-52
[20] Rong-Xia Liu, Min Ye, HongZhu Guo, Kaishun Bi and De-an Guo. Liquid chromatography/eletrospray ionization mass spectrometry for the characterization of twenty-three flavonoids in the extract of Dalbergia odorifera. Rapid Commun. Mass Spectrom. 2005; 19:1557-1565.
[21] Tineke N.Decaestecker, Sofie R.Vande Casteele, Pierre E.Wallemacq, Carlos H. Van Peteghem, Dieter L.Defore and Jan F.Van Bocxlaer. Information-Dependent Acquisition-Mediated LC-MSMS Screening Procedure with Semiquantitative Potential. Anal.Chem. 2004; 76:6365-6373.
[22] Robert Plumb, Jose Castro-Perez, Jennifer granger, Lain Beattie, Karine Joncour and Andrew Wright. Ultra-performance liguid chromatograghy coupled toquadrupole-orthogonal time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 2004;18:2331-2337.
[23] Nuno banderia, Haixu Tang, Vineet Bafna, and Pavel pevzner. Shotgun protein sequencing by tandem mass spectra assembly. Anal.Chem.2004;76:7221-7233.
[24] Wan Chen, Kim-Chung Lee, Ning Liu, Ricky N.S.Wong, Huwei Liu and Zongwei Cai. Liquid chromatography/mass spectrometry for metabonomics investigation of the biochemical effects induced by aristolochic acid in rats: the use of information-dependent acquisition for biomarker identification. Rapid Commun. Mass Spectrom. 2008;22:873-880.
[25] Philip R.Tiller, Sean Yu, Jose CastrPerez, Kerry L.Fillgrove and Thomas A.Baillie. High-through-put,accurate mass liquid chromatography/tandem mass spectrometry on a quadrupole time-of-flight system as a‘first-line’approach for metabolite identification studies. Rapid Commun. Mass Spectrom. 2008;22:1053 -1061.
[26] Robert S.Plumb, Kelly A.Johnson, Paul Rainville, Brian W.Smith, Lan D.Wilson, Jose M.Castro-perez and Jeremy K.Nichlson. UPLC/MSE:a new approach for generating molecular fragment information for biomarker structure elucidation. Rapid Commun. Mass Spectrom. 2006;20:1989-1994.
[27] Kevin P.Bateman, Jose Castro-Perez, Mark Wrona, John P.Shockcor, Kate Yu, Renata Oballa and Deborah A.Nicoll-Griffith. MSE with mass defect filtering for in vitro and in vivo metabolite identification. Rapid Commun. Mass Spectrom. 2007;21:1485-1496.
[28] Dao T.-T.Nguyen, Davy Guillarme, Serge Rudaz, Jean-Luc Veuthey. Chromatographic behaviour and comparison of column packed with sub-2μm stationary phasea in liquid chromatography. Journal of Chromatography A.2006; 1128:105-113.
[29] Chiara Cavaliere, Francesca Cucci, Patrizia Fogilia, Chiara Guarino, Roberto Samperi and Aldo Lagana. Flavonoid profile in soybeans by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2007; 21: 2177-2187.
[30] Robert S.Plumb, Jennifer H.Granger, Chris L.Stumpf, Kelly A.Johnson, BrianW.Smith, Scott Gaulitz, Lan D.Wilson and Jose Castro-Perez. A rapid screening approach to metabonomics using UPLC and oa-TOF mass spectrometry: application to age, gender and diurnal variation in normal/Zucker obese rats and black, white and nude mice. The Analyst.2005; 130:844-849.
[31] Zhuping Wu, Chao Zhang, Chengdui Yang, Xinrong Zhang and Erruo Wu. Simultaneous quantitative determination of norgestrel and progesterone in human serum by high-performance liquid chromatrogrophy-tandem mass spectrometry with atmospheric pressure chemical ionization. The Analyst.2000; 125:2201-2205.
[32] Alexis A.Ramos, Hua Yang, Lauren E. Rosen, and Xudong Yao. Tandem parallel fragmentation of peptides for mass spectrometry. Anal.Chem. 2006;78:6391 -6397.
[33] Michael Frank. Agilent 1200 series rapid resolution LC system and Agilent 6210 TOF MS-Highest data content with highest throughput. Agilent application note.
[34] Mark Wrona, Timo Mauriala, Timo Mauriala, Kevin P.Bateman, Russell J.Mortishire-Smith and Desmond O’Connor.‘All-in-one’analysis for metabolite identification using liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry with collision energy switching. Rapid Commun. Mass Spectrom. 2005; 19:2597-2602.
[35]曾苏.药物代谢学.杭州:浙江大学出版社. 2004;7.
[2]蒋文跃.寻找物质基础非当前中药现代化关键.中草药.2003;34:1-3.
[3]李忠红杜冠华.中药质量监控之我见.世界科学技术-中医药现代化
质量标准研究. 2004;6: 67-71.
[4]耿平博士毕业论文,中国医学科学院协和医科大学. 2007
[5]高莉斯拉甫·艾白,新疆维药与国内外重要质量标准研究的比较.中国民族医药杂志. 2006; 4:29-30.
[6]李斌博士毕业论文,中国医学科学院协和医科大学. 2005
[7]彭涛雍炜安娟等,反相高效液相色谱/质谱法同时测定肉鸡中5种喹诺酮药物残留. Chinese Journal of Analytical Chemistry.2006; 34: 10-14.
[8] Pierluigi Caboni, Giorgia Sarais,Marco Cabras, and Alberto angioni. Determination of 4-Ethylphenol and 4-Ethylguaiacol in Wines by LC-MS-MS and HPLC-DAD-Fluorescence. J.Agric.Food Chem. 2007; 55:7288-7293
[9] Jacek Bielawski, Zdzislaw M. Szulc, Yusuf A. Hannun, Alicja Bielawska. Simultaneous quantitative analysis of bioactive sphingolipids by high-performance liquid chromatography-tandem mass spectrometry. Methods.2006; 39: 82-91.
[10] Gibson GG, Skett P. Pathways of Drug Metabolism. In Introduction to Drug Metabolism, Gibson GG, Skett P(ed.). Blackie Academic and Professional:London,1994;1-34
[11] Mulder GJ, Coughtrie MWH, Burchell B. Glucuronidation. In Conjugation Reactions in Drug Metabolism: an Integrated Approach.Mulder GJ(ed).taylor and Francis: London,1990; 42-91.
[12]陈义,高通微量分析.化学进展Progress in Chemistry. 2005; 17: 573-580.
[13] Charanjit sandhu, Johannes A. Hewel, Gwenael badis, Shaheynoor Taluder, Jian Liu, Timothy R.Hughes, and Andrew Emili. Evaluation of Data-dependent versusTargeted Shotgun Proteomic Approaches for Monitoring Transcription Factor Expression in Breast Cancer.Journal of proteome research. 2008;7:1529-1541.
[14] Geng P, Zhang RP, Aisa HA, He JM, Qu K, Zhu HB, Abliz Z. Fast profiling of the integral metabolism of flavonols in the active fraction of Gossypium herbaceam L. using liquid chromatography/multi-stage tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2007;21: 1877-88
[15]高洁王素娟徐瑞明杨永春张守仁司伊康石建功.以体内吸收分布特点指导花锚成分的研究.药学学报. 2004; 39: 198-203.
[16] Asish B.Chakraborty, Scott J.Berger and John C.Gebler. Use of an integrated MS-multiplexed MS/MS data acquisition strategy for high-coverage peptide mapping studies. Rapid Commun. Mass Spectrom. 2007;21:730-744.
[17] Keyume Ablajan, Zeper Abliz, Xiao-ya Shang, Jiu-ming He, Rui-ping Zhang and Jian-Gong Shi. Structural characterization of flavonol 3,7-di-O-glycosides and determination of the glycosylation position by using negative ion electrospray ionization tandem mass spectrometry. J.Mass Spectrum. 2006; 41:352-360.
[18] P.Kachlicki, J.Einhorn, D.Muth, L.Kerhoas and M.Stobiecki. Evaluation of glycosylation and malonylation patterns in flavonoid glycosides during LC/MS/MS metabolite profiling. J.Mass Spectrum. 2008;43:572-586.
[19] Lucia Politi, Luca Morini and Aldo Polettini. A driect screening procedure for diuretics in human urine by liquid chromatography-tandem mass spectrometry with information depengent acquisition. Clin Chim Acta. 2007;386(1-2):46-52
[20] Rong-Xia Liu, Min Ye, HongZhu Guo, Kaishun Bi and De-an Guo. Liquid chromatography/eletrospray ionization mass spectrometry for the characterization of twenty-three flavonoids in the extract of Dalbergia odorifera. Rapid Commun. Mass Spectrom. 2005; 19:1557-1565.
[21] Tineke N.Decaestecker, Sofie R.Vande Casteele, Pierre E.Wallemacq, Carlos H. Van Peteghem, Dieter L.Defore and Jan F.Van Bocxlaer. Information-Dependent Acquisition-Mediated LC-MSMS Screening Procedure with Semiquantitative Potential. Anal.Chem. 2004; 76:6365-6373.
[22] Robert Plumb, Jose Castro-Perez, Jennifer granger, Lain Beattie, Karine Joncour and Andrew Wright. Ultra-performance liguid chromatograghy coupled toquadrupole-orthogonal time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 2004;18:2331-2337.
[23] Nuno banderia, Haixu Tang, Vineet Bafna, and Pavel pevzner. Shotgun protein sequencing by tandem mass spectra assembly. Anal.Chem.2004;76:7221-7233.
[24] Wan Chen, Kim-Chung Lee, Ning Liu, Ricky N.S.Wong, Huwei Liu and Zongwei Cai. Liquid chromatography/mass spectrometry for metabonomics investigation of the biochemical effects induced by aristolochic acid in rats: the use of information-dependent acquisition for biomarker identification. Rapid Commun. Mass Spectrom. 2008;22:873-880.
[25] Philip R.Tiller, Sean Yu, Jose CastrPerez, Kerry L.Fillgrove and Thomas A.Baillie. High-through-put,accurate mass liquid chromatography/tandem mass spectrometry on a quadrupole time-of-flight system as a‘first-line’approach for metabolite identification studies. Rapid Commun. Mass Spectrom. 2008;22:1053 -1061.
[26] Robert S.Plumb, Kelly A.Johnson, Paul Rainville, Brian W.Smith, Lan D.Wilson, Jose M.Castro-perez and Jeremy K.Nichlson. UPLC/MSE:a new approach for generating molecular fragment information for biomarker structure elucidation. Rapid Commun. Mass Spectrom. 2006;20:1989-1994.
[27] Kevin P.Bateman, Jose Castro-Perez, Mark Wrona, John P.Shockcor, Kate Yu, Renata Oballa and Deborah A.Nicoll-Griffith. MSE with mass defect filtering for in vitro and in vivo metabolite identification. Rapid Commun. Mass Spectrom. 2007;21:1485-1496.
[28] Dao T.-T.Nguyen, Davy Guillarme, Serge Rudaz, Jean-Luc Veuthey. Chromatographic behaviour and comparison of column packed with sub-2μm stationary phasea in liquid chromatography. Journal of Chromatography A.2006; 1128:105-113.
[29] Chiara Cavaliere, Francesca Cucci, Patrizia Fogilia, Chiara Guarino, Roberto Samperi and Aldo Lagana. Flavonoid profile in soybeans by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2007; 21: 2177-2187.
[30] Robert S.Plumb, Jennifer H.Granger, Chris L.Stumpf, Kelly A.Johnson, BrianW.Smith, Scott Gaulitz, Lan D.Wilson and Jose Castro-Perez. A rapid screening approach to metabonomics using UPLC and oa-TOF mass spectrometry: application to age, gender and diurnal variation in normal/Zucker obese rats and black, white and nude mice. The Analyst.2005; 130:844-849.
[31] Zhuping Wu, Chao Zhang, Chengdui Yang, Xinrong Zhang and Erruo Wu. Simultaneous quantitative determination of norgestrel and progesterone in human serum by high-performance liquid chromatrogrophy-tandem mass spectrometry with atmospheric pressure chemical ionization. The Analyst.2000; 125:2201-2205.
[32] Alexis A.Ramos, Hua Yang, Lauren E. Rosen, and Xudong Yao. Tandem parallel fragmentation of peptides for mass spectrometry. Anal.Chem. 2006;78:6391 -6397.
[33] Michael Frank. Agilent 1200 series rapid resolution LC system and Agilent 6210 TOF MS-Highest data content with highest throughput. Agilent application note.
[34] Mark Wrona, Timo Mauriala, Timo Mauriala, Kevin P.Bateman, Russell J.Mortishire-Smith and Desmond O’Connor.‘All-in-one’analysis for metabolite identification using liquid chromatography/hybrid quadrupole time-of-flight mass spectrometry with collision energy switching. Rapid Commun. Mass Spectrom. 2005; 19:2597-2602.
[35]曾苏.药物代谢学.杭州:浙江大学出版社. 2004;7.