Clinical Pharmacology Research Center, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China.
Clinical Pharmacology Research Center, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China.
J Pharm Biomed Anal. 2019 Jun 5;170:112-123. doi: 10.1016/j.jpba.2019.03.034. Epub 2019 Mar 18.
Irinotecan hydrochloride liposome injection (IHLI) is a formulation of anticancer drug irinotecan hydrochloride (CPT-11) entrapped in the aqueous core of liposomes. To understand the pharmacokinetic property and evaluate the relationship between pharmacokinetics and pharmacodynamics/toxicity of IHLI, it is of prime importance to determine the concentrations of free CPT-11, total CPT-11 and its main metabolites (SN-38 and SN-38 G) in human plasma. In this paper, we developed and validated a sensitive and reliable ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to quantify the concentrations of these related substances in human plasma. Free CPT-11, SN-38 and SN-38 G in human plasma were simultaneously separated and extracted by 96-well solid phase extraction (SPE) plate, while total CPT-11 was extracted by protein precipitation (PP) method. The analytes were chromatographed on an Acquity UPLC BEH C column and then detected on a Xevo TQ-S tandem mass spectrometer in multiple reactions monitoring (MRM) mode using positive electrospray ionization (ESI). The UPLC-MS/MS method combined with SPE and PP techniques were fully validated in line with existing guidelines issued by regulatory agencies. In brief, all the analytes achieved a satisfactory selectivity and sensitivity in this method. The calibration curves were proved to be linear over the concentration range of 10-10000 ng/mL for total CPT-11, 0.5-1000 ng/mL for free CPT-11, 0.5-200 ng/mL for SN-38 and SN-38 G, respectively. For all the analytes, the intra- and inter-run precisions were less than 11.4% and the accuracies (in terms of RE%) were within -7.7-7.3% except for accuracies of LLOQs were within -15.8 to 7.2%. Besides, carry-over, extraction recovery, matrix effect, dilution integrity, stability and special matrices were also assessed. Finally, the method was successfully applied to a phase I clinical pharmacokinetic study of IHLI in Chinese subjects with advanced solid tumors.
盐酸伊立替康脂质体注射液（IHLI）是一种将盐酸伊立替康（CPT-11）包封在脂质体水核中的抗癌药物制剂。为了了解 IHLI 的药代动力学特性并评估其药代动力学与药效学/毒性之间的关系，确定人血浆中游离 CPT-11、总 CPT-11 及其主要代谢物（SN-38 和 SN-38G）的浓度至关重要。本文建立并验证了一种灵敏、可靠的超高效液相色谱-串联质谱（UPLC-MS/MS）法，用于定量分析人血浆中这些相关物质的浓度。采用 96 孔固相萃取（SPE）板同时分离和提取人血浆中的游离 CPT-11、SN-38 和 SN-38G，而总 CPT-11 则采用蛋白沉淀（PP）法提取。采用 Acquity UPLC BEH C 柱进行色谱分离，然后在 Xevo TQ-S 串联质谱仪上采用正电喷雾电离（ESI）在多重反应监测（MRM）模式下进行检测。该 UPLC-MS/MS 方法结合 SPE 和 PP 技术，完全符合监管机构发布的现行指南的要求。简而言之，所有分析物在该方法中均达到了令人满意的选择性和灵敏度。总 CPT-11 的校准曲线在 10-10000ng/mL 的浓度范围内、游离 CPT-11 的校准曲线在 0.5-1000ng/mL 的浓度范围内、SN-38 和 SN-38G 的校准曲线在 0.5-200ng/mL 的浓度范围内均呈线性。对于所有分析物，日内和日间精密度均小于 11.4%，准确度（以相对误差[RE%]表示）均在-7.7%至 7.3%范围内，除了定量下限（LLOQ）的准确度在-15.8%至 7.2%范围内。此外，还评估了交叉污染、提取回收率、基质效应、稀释完整性、稳定性和特殊基质。最后，该方法成功应用于中国晚期实体瘤患者 IHLI 的 I 期临床药代动力学研究。
