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超高效液相色谱-串联四极杆飞行时间质谱法和超高效液相色谱-串联三重四极杆质谱法对血浆中苦杏仁苷及其代谢产物苦杏仁素进行定性和定量分析

[Qualitative and quantitative analysis of amygdalin and its metabolite prunasin in plasma by ultra-high performance liquid chromatography-tandem quadrupole time of flight mass spectrometry and ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry].

作者信息

Gao Meng, Wang Yuesheng, Wei Huizhen, Ouyang Hui, He Mingzhen, Zeng Lianqing, Shen Fengyun, Guo Qiang, Rao Yi

出版信息

Se Pu. 2014 Jun;32(6):591-9. doi: 10.3724/sp.j.1123.2014.01021.

Abstract

A method was developed for the determination of amygdalin and its metabolite prunasin in rat plasma after intragastric administration of Maxing shigan decoction. The analytes were identified by ultra-high performance liquid chromatography-tandem quadrupole time of flight mass spectrometry and quantitatively determined by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry. After purified by liquid-liquid extraction, the qualitative analysis of amygdalin and prunasin in the plasma sample was performed on a Shim-pack XR-ODS III HPLC column (75 mm x 2.0 mm, 1.6 microm), using acetonitrile-0.1% (v/v) formic acid aqueous solution. The detection was performed on a Triple TOF 5600 quadrupole time of flight mass spectrometer. The quantitative analysis of amygdalin and prunasin in the plasma sample was performed by separation on an Agilent C18 HPLC column (50 mm x 2.1 mm, 1.7 microm), using acetonitrile-0.1% (v/v) formic acid aqueous solution. The detection was performed on an AB Q-TRAP 4500 triple quadrupole mass spectrometer utilizing electrospray ionization (ESI) interface operated in negative ion mode and multiple-reaction monitoring (MRM) mode. The qualitative analysis results showed that amygdalin and its metabolite prunasin were detected in the plasma sample. The quantitative analysis results showed that the linear range of amygdalin was 1.05-4 200 ng/mL with the correlation coefficient of 0.999 0 and the linear range of prunasin was 1.25-2 490 ng/mL with the correlation coefficient of 0.997 0. The method had a good precision with the relative standard deviations (RSDs) lower than 9.20% and the overall recoveries varied from 82.33% to 95.25%. The limits of detection (LODs) of amygdalin and prunasin were 0.50 ng/mL. With good reproducibility, the method is simple, fast and effective for the qualitative and quantitative analysis of the amygdalin and prunasin in plasma sample of rats which were administered by Maxing shigan decoction.

摘要

建立了一种测定大鼠灌胃给予麻杏石甘汤后血浆中苦杏仁苷及其代谢产物苦杏仁素的方法。采用超高效液相色谱-串联四极杆飞行时间质谱对分析物进行鉴定,并用超高效液相色谱-串联三重四极杆质谱进行定量测定。经液液萃取纯化后,血浆样品中苦杏仁苷和苦杏仁素的定性分析在Shim-pack XR-ODS III高效液相色谱柱(75 mm×2.0 mm,1.6 µm)上进行,流动相为乙腈-0.1%(v/v)甲酸水溶液。检测在Triple TOF 5600四极杆飞行时间质谱仪上进行。血浆样品中苦杏仁苷和苦杏仁素的定量分析在Agilent C18高效液相色谱柱(50 mm×2.1 mm,1.7 µm)上进行,流动相为乙腈-0.1%(v/v)甲酸水溶液。检测在AB Q-TRAP 4500三重四极杆质谱仪上进行,采用电喷雾电离(ESI)接口,在负离子模式和多反应监测(MRM)模式下运行。定性分析结果表明,在血浆样品中检测到了苦杏仁苷及其代谢产物苦杏仁素。定量分析结果表明,苦杏仁苷的线性范围为1.05 - 4200 ng/mL,相关系数为0.9990;苦杏仁素的线性范围为1.25 - 2490 ng/mL,相关系数为0.9970。该方法精密度良好,相对标准偏差(RSD)低于9.20%,总回收率在82.33%至95.25%之间。苦杏仁苷和苦杏仁素的检测限(LOD)为0.50 ng/mL。该方法重现性好,对灌胃给予麻杏石甘汤大鼠血浆样品中苦杏仁苷和苦杏仁素的定性和定量分析简单、快速且有效。

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