A simple, highly sensitive and specific method based on high-performance liquid chromatography (HPLC) with ultraviolet detection was developed for the measurement of venetoclax concentrations in plasma samples. The chromatographic method employed a mobile phase of acetonitrile: 0.5% KH2PO4 (pH 3.5) (80/20, v/v) on a CAPCELL PAK C18 UG120 column at a flow rate of 0.5 mL/min. The quantitative method was validated based on standards described in “Bioanalytical Method Validation: Guidance for Industry” published by the US Food and Drug Administration. The separation of venetoclax and the internal standard R051012 was satisfactory, and the chromatograms were free of interfering peaks from the biological matrix. The intra- and inter-day coefficients of variation for venetoclax assays were <12.9%, whereas intra- and inter-day accuracies were within 13.6%. Only 100 μL of human plasma was required to detect a lower limit of quantification of 10 ng/mL for venetoclax. The recoveries of venetoclax extracted with an Oasis HLB cartridge were between 81 and 85%. The developed HPLC method was successfully applied to the determination of venetoclax concentrations in plasma of acute myeloid leukemia patients taking venetoclax. The degree of drug interactions between venetoclax and CYP3A4 inhibitors can be determined by this HPLC assay.

中文翻译：

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采用紫外检测的高效液相色谱法定量人血浆中的 Venetoclax


开发了一种基于高效液相色谱 (HPLC) 和紫外检测的简单、高灵敏度和特异的方法，用于测量血浆样品中的维奈托克浓度。色谱方法采用乙腈：0.5% KH2PO4 (pH 3.5) (80/20，v/v) 流动相，CAPCELL PAK C18 UG120 柱，流速 0.5 mL/min。定量方法根据美国食品和药物管理局出版的“生物分析方法验证：工业指南”中描述的标准进行验证。 Venetoclax 和内标 R051012 的分离效果令人满意，并且色谱图中没有来自生物基质的干扰峰。 Venetoclax 检测的日内和日间变异系数为 <12.9%，而日内和日间准确度在 13.6% 以内。只需 100 μL 人血浆即可检测 Venetoclax 的定量下限 10 ng/mL。使用 Oasis HLB 柱提取 Venetoclax 的回收率在 81% 至 85% 之间。所开发的高效液相色谱法成功应用于测定服用维奈托克的急性髓系白血病患者血浆中维奈托克浓度。 Venetoclax 和 CYP3A4 抑制剂之间的药物相互作用程度可以通过该 HPLC 测定来确定。