Hydroxychloroquine (HCQ) was originally used as an antimalarial and immunomodulation drug. We developed and validated a simple and sensitive ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous quantitation of HCQ and its three metabolites in rat blood, and reported their pharmacokinetic parameters. The chromatographic separation and detection of analytes were achieved within 4 min on ZORBAX SB-C₈ (3.5 μm, 2.1 × 150 mm) column with gradient elution, and the flow rate was 0.25 mL/min. Simple protein precipitation was successfully applied for sample pretreatment. The HCQ displays a good linearity in the range of 2.0–5000.0 ng/mL, and the three metabolites also show good linearity ranging from 1.0 to 2500.0 ng/mL, with all correlation coefficients (R²) better than 0.98. In conclusion, this rapid, sensitive method was successfully developed, validated, and then applied to a pharmacokinetic study of HCQ in rat model in high dose. The results of the pharmacokinetic study presented an average half-life time 21.14 ± 10.31 h (mean ± SD) of HCQ, which is much shorter in human compared to that in mice. For the three metabolites, longer half-life times (approximately 100 h) were shown in rat.

中文翻译：

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LC-MS/MS 方法测定羟氯喹及其代谢物：在药代动力学研究中的应用

羟氯喹 (HCQ) 最初用作抗疟疾和免疫调节药物。我们开发并验证了一种简单且灵敏的超高效液相色谱-串联质谱 (UHPLC-MS/MS) 方法，用于同时定量大鼠血液中的 HCQ 及其三种代谢物，并报告了它们的药代动力学参数。在 ZORBAX SB-C ₈（3.5  μm, 2.1 × 150 mm) 柱，梯度洗脱，流速为 0.25 mL/min。简单的蛋白质沉淀成功地应用于样品预处理。HCQ在2.0~5000.0 ng/mL范围内表现出良好的线性，三种代谢物在1.0~2500.0 ng/mL范围内也表现出良好的线性，所有相关系数（R ²）均优于0.98。总之，这种快速、灵敏的方法得到了成功开发和验证，然后应用于大鼠模型中高剂量HCQ的药代动力学研究。药代动力学研究结果显示 HCQ 的平均半衰期为 21.14 ± 10.31 小时（平均值 ± SD），与小鼠相比，人的半衰期要短得多。对于这三种代谢物，大鼠的半衰期较长（约 100 小时）。