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液相色谱/串联质谱法快速定量测定人血浆中的加巴喷丁

Rapid quantification of gabapentin in human plasma by liquid chromatography/tandem mass spectrometry.

作者信息

Ramakrishna N V S, Vishwottam K N, Koteshwara M, Manoj S, Santosh M, Chidambara J, Sumatha B, Varma D P

机构信息

Biopharmaceutical Research, Suven Life Sciences Ltd., Serene Chambers, Road # 7, Banjara Hills, Hyderabad 500034, India.

出版信息

J Pharm Biomed Anal. 2006 Feb 13;40(2):360-8. doi: 10.1016/j.jpba.2005.07.012. Epub 2005 Aug 19.

Abstract

A simple, sensitive and rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of gabapentin, a new antiepileptic drug, in human plasma using its structural analogue, 1,1-cyclohexane diacetic acid monoamide (CAM) as internal standard. The method involved a simple protein precipitation by means of acetonitrile followed by a rapid isocratic elution with 10mM ammonium formate buffer/acetonitrile (20/80, v/v, pH 3.0) on Waters Symmetry C(18 reversed phase chromatographic column and analyzed by mass spectrometry in the multiple reaction monitoring mode. The precursor to product ion transitions of m/z 172-->154 and m/z 200-->182 were used to measure the analyte and the IS, respectively. The assay exhibited a linear dynamic range of 40-10000 ng/mL for gabapentin in human plasma. The limit of detection and lower limit of quantification in human plasma were 10 and 40 ng/mL, respectively. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. A run time of 2 min for each sample made it possible to analyze a throughput of more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies.

摘要

建立了一种简单、灵敏、快速的液相色谱/串联质谱(LC-MS/MS)方法,并进行了验证,用于测定人血浆中新型抗癫痫药物加巴喷丁的含量,采用其结构类似物1,1-环己烷二乙酸单酰胺(CAM)作为内标。该方法包括用乙腈进行简单的蛋白沉淀,然后在Waters Symmetry C18反相色谱柱上用10mM甲酸铵缓冲液/乙腈(20/80,v/v,pH 3.0)进行快速等度洗脱,并在多反应监测模式下进行质谱分析。分别用m/z 172→154和m/z 200→182的前体离子到产物离子的转变来测定分析物和内标。该测定法在人血浆中加巴喷丁的线性动态范围为40-10000 ng/mL。人血浆中的检测限和定量下限分别为10和40 ng/mL。在标准曲线范围内的浓度获得了可接受的精密度和准确度。每个样品2分钟的运行时间使得每天能够分析超过400份人血浆样品。该验证方法已成功用于分析人血浆样品,以应用于药代动力学、生物利用度或生物等效性研究。

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