采用超高效液相色谱-串联质谱法同时测定人血浆中左乙拉西坦和加巴喷丁的浓度。

Simultaneous quantification of levetiracetam and gabapentin in plasma by ultra-pressure liquid chromatography coupled with tandem mass spectrometry detection.

机构信息

ARUP Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, Utah, USA.

出版信息

Ther Drug Monit. 2011 Apr;33(2):209-13. doi: 10.1097/FTD.0b013e31820b1fce.

DOI:10.1097/FTD.0b013e31820b1fce

PMID:21297550

Abstract

INTRODUCTION

Gabapentin (Neurontin) and levetiracetam (Keppra) are anticonvulsants with novel structures and suggested therapeutic ranges of 2-10 mg/L and 6-20 mg/L, respectively. Gabapentin is also used extensively to manage neuropathic pain, and for this indication, wherein higher doses are prescribed, plasma concentrations of 15-30 mg/L are typical.

OBJECTIVE

Here, we describe a simple rapid assay to support therapeutic drug monitoring of gabapentin and levetiracetam in plasma by ultra-pressure liquid chromatography couples to tandem mass spectrometry (UPLC-MS/MS) detection.

METHODS

After the addition of internal standard and protein precipitation of patient plasma with methanol:acetonitrile in a 50:50 ratio, 1 μL of supernatant sample is injected onto an Acquity UPLC HSS T3, 1.8 μm, 2.1 × 50 mm (Waters) column. Elution occurs using a linear gradient of acetonitrile and water, each having 0.1% formic acid added. The column is eluted into a Waters Acquity UPLC TQD, operating in a positive mode to detect gabapentin at transition 172.18 > 154.11, levetiracetam at 171.11 > 126, and internal standard (3-amino-2-naphthoic acid) at 188.06 > 170. Secondary transitions for each analyte are also monitored for gabapentin at 172.18 > 137.06, levetiracetam at 171.11 > 154, and internal standard at 188.06 > 115. Runtime is 1.5 minutes per injection with baseline resolved chromatographic separation.

RESULTS

The analytical measurement ranges were 1-150 mg/L for gabapentin and for levetiracetam. Intra-assay imprecision by the coefficient of variance (CV) was less than 8% and interassay CV was less than 5% for both analytes, at 4 different concentrations. Results obtained from patient samples were compared with results generated by established high-performance liquid chromatography-UV methods with the following regression statistics: y = 1.12x - 0.77, r = 0.996, Sy, x = 0.89, and n = 29 for gabapentin and y = 0.991x + 0.70, r = 0.997, Sy, x = 2.24, and n = 30 for levetiracetam. No analytical interferences were identified.

CONCLUSION

: In summary, a simple reliable UPLC-MS/MS method was developed and validated for routine clinical monitoring of gabapentin and levetiracetam.

摘要

简介

加巴喷丁（Neurontin）和左乙拉西坦（Keppra）是具有新颖结构的抗惊厥药物，其治疗范围分别为 2-10mg/L 和 6-20mg/L。加巴喷丁还广泛用于治疗神经病理性疼痛，对于这种需要更高剂量的适应症，典型的血浆浓度为 15-30mg/L。

目的

本文描述了一种简单快速的分析方法，通过超高效液相色谱-串联质谱（UPLC-MS/MS）检测，支持加巴喷丁和左乙拉西坦在血浆中的治疗药物监测。

方法

在加入内标物并以甲醇：乙腈（50:50）比例沉淀患者血浆中的蛋白质后，取 1μL上清液注入 Acquity UPLC HSS T3、1.8μm、2.1×50mm（Waters）柱。采用含有 0.1%甲酸的乙腈和水的线性梯度洗脱。将柱洗脱至 Waters Acquity UPLC TQD，以正模式运行，检测加巴喷丁时的转换 172.18 > 154.11，检测左乙拉西坦时的转换 171.11 > 126，以及内标物（3-氨基-2-萘甲酸）时的转换 188.06 > 170。还监测了每个分析物的次级转换，用于检测加巴喷丁时的转换 172.18 > 137.06，检测左乙拉西坦时的转换 171.11 > 154，以及内标物时的转换 188.06 > 115。每个注射物的运行时间为 1.5 分钟，具有基线分离的色谱分离。

结果

加巴喷丁和左乙拉西坦的分析测量范围均为 1-150mg/L。在 4 个不同浓度下，两种分析物的日内精密度（CV）均小于 8%，日间 CV 均小于 5%。通过与已建立的高效液相色谱-紫外方法生成的结果进行比较，患者样本的结果具有以下回归统计数据：加巴喷丁为 y = 1.12x - 0.77，r = 0.996，Sy，x = 0.89，n = 29，左乙拉西坦为 y = 0.991x + 0.70，r = 0.997，Sy，x = 2.24，n = 30。未发现分析干扰。