UPLC-Q-TOF-MS/MS分析荭草花有效组分在大鼠血清和尿液中的原型成分及其代谢产物
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摘要
应用超高效液相色谱串联电喷雾飞行时间质谱(UPLC-Q-TOF-MS/MS)分析SD大鼠灌胃给予荭草花有效组分后所得血清和尿液中的原型成分及代谢产物。采用Agilent Eclipse Plus C_(18)色谱柱(2. 1 mm×100 mm,1. 8μm),以0. 1%甲酸水(A)-0. 1%甲酸乙腈(B)为流动相进行梯度洗脱。在电喷雾正负离子模式下获取生物样品中化学成分的质谱数据,通过比较空白样品、给药样品和两者的差异图谱确认灌胃后大鼠血清和尿液中原型成分及其代谢产物。利用TOF/MS得到的准确相对分子质量,多级质谱裂解信息,结合对照品及相关文献报道结果对比,对尿液和血清中荭草花原型成分和代谢产物进行鉴定。结果发现,在大鼠血清、尿液中分别检测到12,26个代谢产物(含交叉成分),代谢途径主要包括以氢化还原、开环裂解、羟基化、甲基葡萄糖醛酸化、葡萄糖醛酸化、硫酸化等。该实验建立的方法可靠、有效,可用于荭草花有效部位在血清和尿液中的代谢研究,研究结果为该药材今后的药效物质基础研究提供了理论参考。
Ultra performance liquid chromatography coupled with time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) method was applied to analyze the prototypes and metabolites of the effective components of Polygonum orientale in SD rat serum and urine. The separation was performed on Agilent Eclipse Plus C_(18) column( 2. 1 mm×100 mm,1. 8 μm),with 0. 1% formic acid solution( A)-acetonitrile( B) as the mobile phase for gradient elution. Mass spectrometry data of biological samples were obtained under positive and negative electrospray ion mode. By comparing chromatogram differences between blank samples and drug treatment samples,prototype components and metabolites of the effective components of P. orientale extract were identified. The results showed that 12 metabolites were detected in serum and 26 metabolites in urine( including cross-components) of rats. The main metabolic pathways included hydrogenation,hydroxylation,glucuronidation,sulfation reaction,and methylation-glucuronidation,etc. The method established in this study was reliable and effective for studying the metabolic characteristics of the effective components of P. orientale in rats,and it can provide a reference for further studies on therapeutic material basis of this herb.
Ultra performance liquid chromatography coupled with time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) method was applied to analyze the prototypes and metabolites of the effective components of Polygonum orientale in SD rat serum and urine. The separation was performed on Agilent Eclipse Plus C_(18) column( 2. 1 mm×100 mm,1. 8 μm),with 0. 1% formic acid solution( A)-acetonitrile( B) as the mobile phase for gradient elution. Mass spectrometry data of biological samples were obtained under positive and negative electrospray ion mode. By comparing chromatogram differences between blank samples and drug treatment samples,prototype components and metabolites of the effective components of P. orientale extract were identified. The results showed that 12 metabolites were detected in serum and 26 metabolites in urine( including cross-components) of rats. The main metabolic pathways included hydrogenation,hydroxylation,glucuronidation,sulfation reaction,and methylation-glucuronidation,etc. The method established in this study was reliable and effective for studying the metabolic characteristics of the effective components of P. orientale in rats,and it can provide a reference for further studies on therapeutic material basis of this herb.
引文
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[21]潘玄玄,宋粉云,林秀莲,等.基于UPLC-Q-TOF-MS分析沉香化气丸在大鼠体内的代谢产物[J].中国中药杂志,2018,43(3):527.
[2]谢玉敏,杨武,陆苑,等.荭草花提取物中5种成分在大鼠体内的肠吸收动力学[J].中成药,2016,38(4):770.
[3]李月婷,胡杰,谢玉敏,等.荭草花不同制备工艺样品对犬急性心肌缺血的保护作用[J].中药材,2015,38(1):115.
[4]向文英,梅朝叶,孙慧园,等.基于UHPLC-Q-TOF-MS分析荭草花提取物在H9c2心肌细胞中效应物质[J].中国实验方剂学杂志,2017,23(12):90.
[5]李勇军,何迅,刘志宝,等.荭草花化学成分的研究[J].中国中药杂志,2009,34(20):2613.
[6]郑林,杨武,向文英,等. UPLC-MS/MS同时测定静脉注射荭草花提取物大鼠血浆中3种成分及其药代动力学研究[J].中药材,2016,39(7):1574.
[7]黄勇,胡杰,陆苑,等. UPLC-MS法测定大鼠血浆中荭草花的3个指标成分及其药动学研究[J].中国药理学通报,2015,31(9):1314.
[8]梅晓丹,王喻淇,王子健,等.丹参素在大鼠体内代谢产物的鉴定分析[J].中国中药杂志,2018,43(19):3933.
[9]申文雯,张银,仇守蓓,等.蒸三七皂苷类活性组分在大鼠体内代谢产物鉴定的研究[J].中国中药杂志,2017,42(20):3996.
[10]张丹,王彩虹,金滢,等.高效液相色谱-高分辨质谱法研究药物代谢产物的数据处理策略[J].质谱学报,2017,38(4):410.
[11]张晶,温泉,徐旭,等.基于UHPLC-QTOF-MS/MS技术的铁筷子中3个香豆素在大鼠体内代谢研究[J].中国中药杂志,2018,43(21):4330.
[12] Sleno L. The use of mass defect in modern mass spectrometry[J].J Mass Spectrom,2012,47:226.
[13]覃小丽,孙慧园,杨武,等.UPLC-ESI-Q-TOF-MS/MS分析槲皮苷在大鼠肠道菌群中的代谢[J].中国中药杂志,2017,42(2):357.
[14] Cao W,Wang X,Li H,et al. Studies on metabolism of total glucosides of paeony from Paeoniae Radix Alba in rats by UPLC-QTOF-MS/MS[J]. Biomed Chromatogr,2015,29:1769.
[15] Ma H,Liu Y,Mai X,et al. Identification of the constituents and metabolites in rat plasma after oral administration of Huanglian Shangqing Pills by ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry[J]. J Pharm Biomed Anal,2016,125:194.
[16] Zhu F,Chen J,Wang H,et al. Analysis of the chemical constituents and rats metabolites after oral administration of Nauclea officinalis by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry[J]. J Chromatogr B,2015,1007:54.
[17] Lecompte Y,Rosset M,Richeva C,et al. UPLC-ESI-Q-TOFMSEidentification of urinary metabolites of the emerging sport nutrition supplement methoxyisoflavone in human subjects[J]. J Pharm Biomed Anal,2014,96:127.
[18] Yang Y,Zhao X Y,Pan Y,et al. Identification of the chemical compositions of Ponkan peel by ultra performance liquid chromatography coupled with quadrupole time of-flight mass spectrometry[J]. Analmethods,2016,8:893.
[19] Wang H P,Chen C,Liu Y,et al. Metabolic profile of Yixinshu Capsule in rat by ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry analysis[J]. Rsc Adv,2015,5:80583.
[20] Zhu F,Chen J,Wang H,et al. Analysis of the chemical constituents and rats metabolites after oral administration of Nauclea officinalis by ultra-performance liquid chromatography quadrupole timeof-flight mass spectrometry[J]. J Chromatogr B 2015,1007:54.
[21]潘玄玄,宋粉云,林秀莲,等.基于UPLC-Q-TOF-MS分析沉香化气丸在大鼠体内的代谢产物[J].中国中药杂志,2018,43(3):527.