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采用高效液相色谱-紫外检测法同时测定人血浆中马拉维若和拉替拉韦的含量。

Simultaneous determination of maraviroc and raltegravir in human plasma by HPLC-UV.

作者信息

Notari Stefania, Tommasi Chiara, Nicastri Emanuele, Bellagamba Rita, Tempestilli Massimo, Pucillo Leopoldo Paolo, Narciso Pasquale, Ascenzi Paolo

机构信息

Istituto Nazionale per le Malattie Infettive IRCCS Lazzaro Spallanzani, Roma, Italy.

出版信息

IUBMB Life. 2009 Apr;61(4):470-5. doi: 10.1002/iub.181.

Abstract

Therapeutic drug monitoring is pivotal to improve the management of HIV infection. Here, a new HPLC-UV method to quantify simultaneously maraviroc and raltegravir levels in human plasma is reported. Remarkably, this is the first method for maraviroc determination in human plasma. The volume of the plasma sample was 600 microL. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (30 mg divinylbenzene and N-vinylpyrrolidone) and evaporation in a water bath under nitrogen stream. The extracted samples were reconstituted with 200 microL 50/50 of mobile-phase solution (0.01 M KH(2)PO(4) and acetonitrile). Twenty microliters of these samples were injected into a HPLC-UV system, the analytes were eluted on an analytical dC18 Atlantis column (150 mm x 4.6 mm I.D.) with a particle size of 5 microm. The mobile phase (0.01 M KH(2)PO(4) and acetonitrile) was delivered at 1.0 mL/min with isocratic elution. The total run time for a single analysis was 10 min; maraviroc and raltegravir were detected by UV at 197 and 300 nm. The calibration curves were linear up to 2,500 ng/mL. The absolute recovery ranged between 93 and 100%. The HPLC-UV method reported here has been validated and is currently applied to monitor plasma levels of maraviroc and raltegravir in HIV-infected patients.

摘要

治疗药物监测对于改善HIV感染的管理至关重要。本文报道了一种新的HPLC-UV方法,可同时定量测定人血浆中马拉维若和拉替拉韦的水平。值得注意的是,这是第一种用于测定人血浆中马拉维若的方法。血浆样本体积为600微升。该方法包括使用Oasis HLB Cartridge 1 cc(30毫克二乙烯基苯和N-乙烯基吡咯烷酮)进行自动固相萃取,并在氮气流下于水浴中蒸发。提取的样品用200微升50/50的流动相溶液(0.01 M KH₂PO₄和乙腈)复溶。将20微升这些样品注入HPLC-UV系统,分析物在粒径为5微米的分析型dC18 Atlantis柱(150毫米×4.6毫米内径)上洗脱。流动相(0.01 M KH₂PO₄和乙腈)以1.0毫升/分钟的流速等度洗脱。单次分析的总运行时间为10分钟;马拉维若和拉替拉韦在197和300纳米处通过紫外检测。校准曲线在高达2500纳克/毫升的范围内呈线性。绝对回收率在93%至100%之间。本文报道的HPLC-UV方法已经过验证,目前用于监测HIV感染患者血浆中马拉维若和拉替拉韦的水平。

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