Istituto Nazionale per le Malattie Infettive IRCCS, Lazzaro Spallanzani, Roma, Italy.
IUBMB Life. 2013 Sep;65(9):800-5. doi: 10.1002/iub.1197. Epub 2013 Jul 29.
Telaprevir is a direct acting antiviral agent, used with pegylated interferon and ribavirin for the management of chronic hepatitis C virus (HCV) genotype 1 infection, in patients not responding to therapy with pegylated interferon and ribavirin only. Although 75% of patients achieve a sustained virological response after treatment with telaprevir, adverse drug-drug interactions and undesirable events often occur. Therefore, telaprevir monitoring is pivotal to improve the management of HCV infection. Here, the first High-Performance Liquid Chromatography-Ultraviolet (HPLC-UV) method to quantify telaprevir in human plasma of HCV-genotype 1-infected patients is reported. The volume of the plasma sample was 700 μL. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (divinylbenzene and N-vinylpyrrolidone). The extracted samples were reconstituted with 150 μL of 60/40 water/acetonitrile. Thirty microliters of these samples was injected into a HPLC-UV system, and the analytes were eluted on a X Terra(®) RP18 column (250 mm × 4.6 mm i.d.) with a particle size of 5 μm. The mobile phase (ammonium acetate buffer, 150 mM, pH 8.0, and methanol:acetonitrile 50:50) was delivered at 1.0 mL/min with linear gradient elution. The total run time for a single analysis was 16 min; telaprevir was detected by UV at 276 and 286 nm. The calibration curve was linear from 312.5 to 20,000 ng/mL (r(2) > 0.996). The absolute recovery of telaprevir ranged between 89 and 93% at concentrations of quality control samples of 800, 4,000, 8,000, and 16,000 ng/mL. Both precision and accuracy were always <15%. The HPLC-UV method reported here: (i) has been validated, (ii) is currently applied to monitor telaprevir in plasma of HCV-infected patients, and (iii) appears useful in a routine laboratory. ,
替拉瑞韦是一种直接作用的抗病毒药物，与聚乙二醇干扰素和利巴韦林联合用于治疗慢性丙型肝炎病毒（HCV）基因型 1 感染，用于对聚乙二醇干扰素和利巴韦林单独治疗无应答的患者。尽管 75%的患者在接受替拉瑞韦治疗后可获得持续病毒学应答，但常发生药物相互作用和不良事件。因此，替拉瑞韦监测对于改善 HCV 感染的管理至关重要。这里，报道了一种用于检测 HCV 基因型 1 感染患者人血浆中替拉瑞韦的首种高效液相色谱-紫外（HPLC-UV）法。血浆样品体积为 700μL。该方法涉及用 Oasis HLB 1 cc （二乙烯基苯和 N-乙烯基吡咯烷酮）自动固相萃取。提取的样品用 150μL 60/40 水/乙腈重新溶解。将 30μL 这些样品注入 HPLC-UV 系统，分析物在 X Terra(®) RP18 柱（250mm×4.6mm i.d.，粒径 5μm）上洗脱。流动相（150mM 乙酸铵缓冲液，pH8.0，甲醇/乙腈 50/50）以 1.0mL/min 的线性梯度洗脱输送。单次分析的总运行时间为 16min；替拉瑞韦在 276nm 和 286nm 处用 UV 检测。校准曲线在 312.5 至 20,000ng/mL 之间呈线性（r(2)＞0.996）。替拉瑞韦在 800、4,000、8,000 和 16,000ng/mL 浓度的质控样品中的绝对回收率在 89%至 93%之间。精密度和准确度始终＜15%。这里报道的 HPLC-UV 方法：（i）已经过验证，（ii）目前用于监测 HCV 感染患者的替拉瑞韦血浆浓度，（iii）在常规实验室中似乎有用。
