LC-MS/MS法同时测定大鼠血浆中6种独一味成分
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摘要
目的:开发建立独一味药材中芹菜素、木犀草素、木犀草苷、绿原酸、毛蕊花糖苷和连翘酯苷B6种成分在大鼠血浆中的同步定量分析方法。方法:采用蛋白沉淀法进行样品前处理,选用安捷伦ZORBAX SB-C18(2.1mm×150mm i.d.,3.5μm)色谱柱及梯度洗脱法进行色谱分离,运用液相色谱串联质谱(LC-MS/MS)技术在多重反应监测(MRM)模式下进行样品测定,并从专属性、线性范围、准确度、精密度、回收率、介质效应和稳定性等方面对方法学进行了系统的验证。结果:在13min的分析时间内,能够实现对6种目标化合物的准确定量,方法学验证中的各项指标均符合生物样品分析的要求。结论:所建立的分析方法准确、灵敏、可靠,可用于研究独一味药材给药后相应成分在大鼠血浆中的暴露情况,为药材的质量分析和控制提供科学依据,也为该药材体内过程和效应机制的研究奠定了基础。
Objective: To develop and establish a quantitation method for the simultaneous determination of apigenin, luteolin, cynaroside, chlorogenic acid, verbascoside and forsythoside B fromLamiophlomis rotata in rat plasma. Methods: Simple protein precipitation was used for sample pre-treatment, an Agilent ZORBAX SB-C18(2.1 mm×150 mm i.d., 3.5μm) column and a gradient elution method were chosen for chromatographic separation, tandem mass spectrometer(LC-MS/MS) technique using multiple reaction monitoring mode(MRM) was applied for determination; a systematic method validation was performed through specificity, linear range, accuracy, precision, recovery, matrix effect and stability. Results: Accurate quantitation of the 6 components was realized within an analytical run time of 13 min, and in method validation, each index met the required criteria of bio-sample analysis. Conclusion: The analytical method established was accurate, precise and reliable, which could be used to reveal the plasma exposure after rat administration of Lamiophlomis rotata, provide further scientific evidence to its quality analysis and control, and also promote the physiological disposition and pharmacodynamic mechanism study of this medicinal material.
Objective: To develop and establish a quantitation method for the simultaneous determination of apigenin, luteolin, cynaroside, chlorogenic acid, verbascoside and forsythoside B fromLamiophlomis rotata in rat plasma. Methods: Simple protein precipitation was used for sample pre-treatment, an Agilent ZORBAX SB-C18(2.1 mm×150 mm i.d., 3.5μm) column and a gradient elution method were chosen for chromatographic separation, tandem mass spectrometer(LC-MS/MS) technique using multiple reaction monitoring mode(MRM) was applied for determination; a systematic method validation was performed through specificity, linear range, accuracy, precision, recovery, matrix effect and stability. Results: Accurate quantitation of the 6 components was realized within an analytical run time of 13 min, and in method validation, each index met the required criteria of bio-sample analysis. Conclusion: The analytical method established was accurate, precise and reliable, which could be used to reveal the plasma exposure after rat administration of Lamiophlomis rotata, provide further scientific evidence to its quality analysis and control, and also promote the physiological disposition and pharmacodynamic mechanism study of this medicinal material.
引文
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[15]Chen Jing,Wang Yang,Liang Xinlei,et al.Simultaneous determination of shanzhiside methyl ester,8-O-acetylshanzhiside methyl ester and luteolin-7-O-?-D-glucopyranoside in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry and its application to a pharmacokinetic study after oral administration of Lamiophlomis rotata Pill.Journal of Chromatography B,2016,1020:62-66
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[18]Wu Liang,Li Lin,Wang Meng,et al.Target and non-target identification of chemical components in Lamiophlomis rotata by liquid chromatography/quadrupole time-of-flight mass spectrometry using a three-step protocol.Rapid Communications in Mass Spectrometry,2016,30(19):2145-2154
[2]关枫,王艳宏.藏药独一味药理作用及临床应用新进展.世界科学技术-中医药现代化,2007,9(2):104-110
[3]张凤,孙连娜,陈万生.独一味的化学成分及药理作用.药学实践杂志,2008,26(3):169-171
[4]林天慕,顾宜,方坤泉,等.藏药独一味两种方法提取组分对小鼠镇痛作用的影响.第四军医大学学报,2003,24(5):444-445
[5]贾正平,李茂星,张汝学,等.独一味止血有效部位的实验研究.解放军药学学报,2005,21(4):272-274
[6]李茂星,贾正平,胡之德,等.藏药独一味止血有效部位总环烯醚萜苷对大鼠血液凝集参数的影响.中国药房,2007,18(3):231-233
[7]贾正平,李茂星,张汝学,等.独一味抗肿瘤活性成分的体外筛选.西北国防医学杂志,2005,26(3):173-175
[8]邱峰.浅谈中药成分体内代谢研究.国际药学研究杂志,2010,37(5):321-238
[9]刘昌孝.我国药代动力学研究发展的回顾.中国药学杂志,2010,45(2):81-89
[10]Wu Liang,Hao Hai Ping,Wang Guang Ji.LC/MS based tools and strategies on qualitative and quantitative analysis of herbal components in complex matrixes.Curr Drug Metab,2012,13(9):1251-1265
[11]屠鹏飞,史社坡,姜勇.中药物质基础研究思路与方法.中草药,2012,43(2):209-215
[12]乔雪,果德安,叶敏.中药体内代谢研究的思路与方法.世界科学技术-中医药现代化,2014,16(3):532-537
[13]邹慧龙,王双虎,周云芳.UPLC-MS/MS测定大鼠血浆中8-O-乙酰山栀苷甲酯浓度.中国现代应用药学,2016,33(4):440-444
[14]陈健苗,王双虎,丁汀.8-O-乙酰山栀苷甲酯和山栀苷甲酯在大鼠体内的药代动力学分析.中国实验方剂学杂志,2016,22(8):100-104
[15]Chen Jing,Wang Yang,Liang Xinlei,et al.Simultaneous determination of shanzhiside methyl ester,8-O-acetylshanzhiside methyl ester and luteolin-7-O-?-D-glucopyranoside in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry and its application to a pharmacokinetic study after oral administration of Lamiophlomis rotata Pill.Journal of Chromatography B,2016,1020:62-66
[16]桑育黎,郝延军.藏药独一味化学成分及测定方法研究进展.中草药,2006,37(10):附9-附10
[17]吴亮,徐立,李璘,等.不同提取溶剂对独一味溶出成分的影响.中药材,2016,39(3):571-574
[18]Wu Liang,Li Lin,Wang Meng,et al.Target and non-target identification of chemical components in Lamiophlomis rotata by liquid chromatography/quadrupole time-of-flight mass spectrometry using a three-step protocol.Rapid Communications in Mass Spectrometry,2016,30(19):2145-2154