超高效液相色谱-四极杆飞行时间质谱法测定大鼠血浆中的N6-羟苄腺苷

doi:10.3724/SP.J.1123.2015.03017

色谱 ›› 2015, Vol. 33 ›› Issue (7): 699-703.DOI: 10.3724/SP.J.1123.2015.03017

超高效液相色谱-四极杆飞行时间质谱法测定大鼠血浆中的N⁶-羟苄腺苷

唐春兰^1,2, 王莉¹, 程孟春¹, 刘欣欣¹, 肖红斌¹

1. 中国科学院大连化学物理研究所, 中国科学院分离分析化学重点实验室, 辽宁大连 116023;
2. 中国科学院大学, 北京 100049

收稿日期:2015-03-12 出版日期:2015-07-08 发布日期:2015-07-27
通讯作者: 肖红斌
基金资助:
国家自然科学基金项目(81001629);重大新药创制专项课题(2014XZ09304-307).

Determination of N⁶-(4-hydroxybenzyl) adenine riboside in rat plasma by ultra performance liquid chromatography- quadrupole time of flight mass spectrometry

TANG Chunlan^1,2, WANG Li¹, CHENG Mengchun¹, LIU Xinxin¹, XIAO Hongbin¹

1. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2015-03-12 Online:2015-07-08 Published:2015-07-27

摘要/Abstract

摘要：

建立了测定大鼠血浆中N⁶-羟苄腺苷的超高效液相色谱-四极杆飞行时间质谱(UPLC-QTOF-MS)分析方法,并应用于N⁶-羟苄腺苷药代动力学研究。采用Agilent ZORBAX SB-C18色谱柱(150 mm×3 mm, 1.8 μm)对目标物分离,以0.2%(v/v)甲酸水溶液和乙腈为流动相进行梯度洗脱;以6-苄氨基嘌呤作为内标,在电喷雾离子源正离子模式下进行定量分析。结果表明:N⁶-羟苄腺苷在0.625~160 ng/mL的范围内线性关系良好(r > 0.99);方法的回收率在88.41%~108.26%之间;检出限达到0.1 ng/mL;日内和日间准确度(以RE计,RE=(测定浓度-加标浓度)/加标浓度×100%)均在±15%之内,精密度(以RSD计)均小于6%。该方法选择性好,灵敏度高,重现性好,结果准确,可用于N⁶-羟苄腺苷的血药浓度监测及药代动力学研究。

关键词: N⁶-羟苄腺苷, 超高效液相色谱-四极杆飞行时间质谱, 大鼠血浆, 药代动力学

Abstract:

An ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) method was developed and validated for the determination of N⁶-(4-hydroxybenzyl) adenine riboside and its pharmacokinetics in rat plasma. The chromatographic conditions were optimized. The separation was performed on an Agilent ZORBAX SB-C18 column(150 mm×3 mm, 1.8 μm) with a gradient elution of 0.2%(v/v) formic acid aqueous solution and acetonitrile as the mobile phases at a flow rate of 0.35 mL/min. The detection was accomplished in positive mode with electrospray ionization (ESI) by UPLC-QTOF-MS, and 6-benzylamino purine was used as the internal standard (IS). The results showed that the linear range of calibration curve was 0.625-160 ng/mL for N⁶-(4-hydroxybenzyl) adenine riboside in rat plasma with the correlation coefficient more than 0.99. The recoveries were 88.41%-108.26%. The limit of detection was 0.1 ng/mL. The intra-day and inter-day precisions (RSDs) were less than 6%, and intra-day and inter-day accuracies (REs, RE=(measured concentration-spiked concentration)/spiked concentration×100%) were less than ±15%. The method is rapid, sensitive and accurate for the quantitation of N⁶-(4-hydroxybenzyl) adenine riboside, which can be used for the study of pharmacokinetics of N⁶-(4-hydroxybenzyl) adenine riboside.

Key words: N⁶-(4-hydroxybenzyl) adenine riboside, pharmacokinetics, rat plasma, ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF-MS)

中图分类号:

O658

唐春兰, 王莉, 程孟春, 刘欣欣, 肖红斌. 超高效液相色谱-四极杆飞行时间质谱法测定大鼠血浆中的N⁶-羟苄腺苷[J]. 色谱, 2015, 33(7): 699-703.

TANG Chunlan, WANG Li, CHENG Mengchun, LIU Xinxin, XIAO Hongbin. Determination of N⁶-(4-hydroxybenzyl) adenine riboside in rat plasma by ultra performance liquid chromatography- quadrupole time of flight mass spectrometry[J]. Chinese Journal of Chromatography, 2015, 33(7): 699-703.

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超高效液相色谱-四极杆飞行时间质谱法测定大鼠血浆中的N⁶-羟苄腺苷

Determination of N⁶-(4-hydroxybenzyl) adenine riboside in rat plasma by ultra performance liquid chromatography- quadrupole time of flight mass spectrometry

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