HPLC-ESI-MSn分析葡萄籽原花青素在人体尿液中的代谢产物
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摘要
目的采用高效液相-电喷雾离子阱质谱联用仪(HPLC-ESI-MSn)鉴定人体尿液中葡萄籽原花青素(Grape Seed Procyanidin,GSP)的代谢产物。方法招募身体健康的受试者口服GSP胶囊,采集受试者0-4h和17-24h内尿样,混合后经乙酸乙酯液-液萃取和C18固相萃取富集净化目标代谢产物。再采用HPLC-ESI-MSn分析代谢产物,确定代谢产物种类。结果该研究共在人体尿液中发现23种GSP代谢产物,其中儿茶素和表儿茶素Ⅱ相代谢产物8种,羟基苯戊内脂类及其Ⅱ相代谢产物8种,酚酸和芳香酸7种。结论该研究采用纯度大于95%的GSP进行人群试食实验,避免了其他成分对实验结果的干扰,可为人体尿液中GSP代谢产物的测定提供参考。
Objective To identify the main metabolites of grape seed procyanidin in human urine by high performance liquid-electrospray ionization mass spectrometry detectorion trap(HPLC-ESI-MSn).Methods Healthy subjects were recruited,and the 0-4 h and 17-24 h urine samples of the subjects after oral giving proanthocyandins capsule were collected.After mixing their urine,it was condensed and purified by liquid-liquid extraction and solid phase extraction.Then the extract solution was analyzed by HPLC-ESI-MSn,so as to identify the kinds of metabolites.Results 23 kinds of metabolites were identified,including 8 kinds of phase Ⅱ metabolites of catechin and epicatechin,8 kinds of hydroxyphenylvalerolactones,7 kinds of aromatic acids and phenlic acids.Conclusion In this study,the trials was performed by grape seed procyanidin with a purity more than 95% in order to eliminate the interference caused by impurity on the experimental results,therefore the results provide some references for the determination of grape seed procyanidin metabolites in human urine.Conclusion In this study,the trials was performed by grape seed procyanidin with a purity more than 95% in order to eliminate the interference caused by impurity on the experimental results,therefore the results provide some references for the determination of grape seed procyanidin metabolites in human urine.
Objective To identify the main metabolites of grape seed procyanidin in human urine by high performance liquid-electrospray ionization mass spectrometry detectorion trap(HPLC-ESI-MSn).Methods Healthy subjects were recruited,and the 0-4 h and 17-24 h urine samples of the subjects after oral giving proanthocyandins capsule were collected.After mixing their urine,it was condensed and purified by liquid-liquid extraction and solid phase extraction.Then the extract solution was analyzed by HPLC-ESI-MSn,so as to identify the kinds of metabolites.Results 23 kinds of metabolites were identified,including 8 kinds of phase Ⅱ metabolites of catechin and epicatechin,8 kinds of hydroxyphenylvalerolactones,7 kinds of aromatic acids and phenlic acids.Conclusion In this study,the trials was performed by grape seed procyanidin with a purity more than 95% in order to eliminate the interference caused by impurity on the experimental results,therefore the results provide some references for the determination of grape seed procyanidin metabolites in human urine.Conclusion In this study,the trials was performed by grape seed procyanidin with a purity more than 95% in order to eliminate the interference caused by impurity on the experimental results,therefore the results provide some references for the determination of grape seed procyanidin metabolites in human urine.
引文
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[12] 林少美,陈珞洛,王超,等.电感耦合等离子质谱法测定香菇中的20种元素[J].中国卫生检验杂志,2014,24(5):670-673.
[13] KATHERYN M,GOODRICH,ANDREW P.UPLC-MS/MS analysis of native catechins and procyanidins and their microbial metabolites in intestinal contentsand tissues of male Wistar Furth inbred rats[J].Journal of Chromatography B,2014(958):63-74.
[14] MARIA M,ZARA P,FRANCISCA I,et al.Plasma kinetics and microbial biotransformation of grape seed flavanols in rats[J].Journal of functional foods,2015,12(28):478-488.
[15] 李可,丘汾.免疫亲和层析-超高效液相色谱法测定大米中4 种黄曲霉毒素[J].中国卫生检验杂志,2014,24(16):2328-2330.
[16] LI SY,SUI Y,XIAO J.Absorption and urinary excretion of A-type procyanidin oligomers from Litchi chinensis pericarp in rats by selected ion monitoring liquid chromatography-mass spectrometry[J].Food Chemistry,2013,138(3):1536-1542.
[17] 张东明.酚酸化学[M].第1版.北京:化学工业出版社,2009:64-65.
[18] MIREIA U,MARIA M,NASIRUDDIN K.Targeted metabolic profiling of phenolics in urine and plasma after regular consumption of cocoa by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2009,1216(43):7258-7267.
[19] JAVIER I,OTTAVIANIA,TONY Y,et al.Structurally related(-)-epicatechin metabolites in humans Assessment using denovo chemically synthesized authentic standards[J].Free Radical Biology and Medicine,2012,52(8):1403-1412.
[20] MARIA M,ZARA P,BEGONA M,et al.A Rapid Method to Determine Colonic Microbial Metabolites Derived from Grape Flavanols in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry[J].Journal of Agricultural and Food Chemistry,2014,62(31):7698-7706.
[2] HACKMAN R M,POLAGRUTI J A,ZHU Q Y,et al.Flavanols:digestion absorption and bioactivity[J].Phytochem.Rev,2008,13(7):195-208.
[3] XU J,RONG S,XIE B,et al.Procyanidins extracted from the lotus seedpod ameliorate age-related antioxidant deficit in aged rats[J].The Journals of Gerontology Series A:Biological Sciences and Medical Sciences,2010,65(3):236-241.
[4] BEL L,DONOYA N,WONG R,et al.(+)-Catechin in human plasma after ingestion of a single serving of reconstituted red wine[J].The American Journal of Clinical Nutrition,2000,32(71):103-108.
[5] KEQI N O,LI W G.Absorption and metabolism of proanthocyanidins[J].Journal of functional foods,2014,7(9):43-53.
[6] ROMANOY M F,VITON F,FUMEAUX R,et al.Epicatechin B-ring conjugates:First enantioselective synthesis and evidence for their occurrence in human biological fluids[J].Organic Letters,2012,14(8):3902-3905.
[7] OU K,PERCIVAL S,ZOU T,et al.Transport of cranberry A-type procyanidin dimers trimers and tetramers across monolayers of human intestinal epithelial Caco-2 cells[J].Journal of Agricultural and Food Chemistry,2012,60(7):1390-1396.
[8] STOUPI S,WILLIAMSON G,VITON F,et al.In vivo bioavailability absorption excretion and pharmacokinetics of [14C] procyanidin B2 in male rats[J].Drug Metabolism and Disposition,2010,38(8):287-291.
[9] 王同蕾,任硕.不同消解方法-石墨炉原子吸收光谱法测定小麦中铅、镉含量的比较[J].中国卫生检验杂志,2016,26(22):3342-3344.
[10] 朱文进,章小雨,王辉.快速消解-石墨炉原子吸收光谱法测定土壤中镉[J].中国卫生检验杂志,2017,27(6):789-791.
[11] 李颖,孙珏,李峰.高效液相色谱-串联质谱法同时测定鱼、虾中18 种抗生素药物残留[J].中国卫生检验杂志,2018,28(23):2830-2837.
[12] 林少美,陈珞洛,王超,等.电感耦合等离子质谱法测定香菇中的20种元素[J].中国卫生检验杂志,2014,24(5):670-673.
[13] KATHERYN M,GOODRICH,ANDREW P.UPLC-MS/MS analysis of native catechins and procyanidins and their microbial metabolites in intestinal contentsand tissues of male Wistar Furth inbred rats[J].Journal of Chromatography B,2014(958):63-74.
[14] MARIA M,ZARA P,FRANCISCA I,et al.Plasma kinetics and microbial biotransformation of grape seed flavanols in rats[J].Journal of functional foods,2015,12(28):478-488.
[15] 李可,丘汾.免疫亲和层析-超高效液相色谱法测定大米中4 种黄曲霉毒素[J].中国卫生检验杂志,2014,24(16):2328-2330.
[16] LI SY,SUI Y,XIAO J.Absorption and urinary excretion of A-type procyanidin oligomers from Litchi chinensis pericarp in rats by selected ion monitoring liquid chromatography-mass spectrometry[J].Food Chemistry,2013,138(3):1536-1542.
[17] 张东明.酚酸化学[M].第1版.北京:化学工业出版社,2009:64-65.
[18] MIREIA U,MARIA M,NASIRUDDIN K.Targeted metabolic profiling of phenolics in urine and plasma after regular consumption of cocoa by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2009,1216(43):7258-7267.
[19] JAVIER I,OTTAVIANIA,TONY Y,et al.Structurally related(-)-epicatechin metabolites in humans Assessment using denovo chemically synthesized authentic standards[J].Free Radical Biology and Medicine,2012,52(8):1403-1412.
[20] MARIA M,ZARA P,BEGONA M,et al.A Rapid Method to Determine Colonic Microbial Metabolites Derived from Grape Flavanols in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry[J].Journal of Agricultural and Food Chemistry,2014,62(31):7698-7706.