Quantification of hydroxychloroquine (HCQ) and its two metabolites desethylchloroquine and desethylhydroxychloroquine in human blood can provide insight into the pharmacokinetic/pharmacodynamic characteristics of HCQ for the treatment of systemic lupus erythematosus (SLE), which is crucial for the optimization of the therapy. A simple, sensitive and optimized high performance liquid chromatography with fluorescence detection method has been developed and validated for the simultaneous determination of HCQ and its two metabolites in human blood. After addition of internal standard chloroquine, the blood sample was deproteinized with 2-fold acetonitrile and separated on an YMC-Triart C18 column (250 × 4.6 mm, 5 μm) with a mobile phase of 20 mM sodium phosphate buffer solution containing 0.25% triethylamine (pH 8.0)—acetonitrile (60:40, v/v). The analytes were detected by using fluorescence detection at an excitation and emission wavelength of 337 and 405 nm, respectively. The method was linear over the range of 3–3000 ng/mL for all three analytes and the chromatographic run time was 9 min. The values for intra- and inter-day precisions were ranged from 1.3 to 7.3. This method was successfully applied to quantify the concentrations of HCQ and its two metabolites in blood of 92 SLE patients.

中文翻译：

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一种用于同时定量系统性红斑狼疮患者血液中羟氯喹及其两种代谢物的灵敏且优化的 HPLC-FLD 方法。

人血中羟氯喹 (HCQ) 及其两种代谢物去乙基氯喹和去乙基羟氯喹的定量可以深入了解 HCQ 治疗系统性红斑狼疮 (SLE) 的药代动力学/药效学特征，这对于优化治疗至关重要。开发并验证了一种简单、灵敏且经过优化的高效液相色谱荧光检测方法，可用于同时测定人血中的 HCQ 及其两种代谢物。加入内标氯喹后，血样用2倍乙腈脱蛋白，在YMC-Triart C18柱（250×4.6 mm，5 μm）上分离，流动相为含0.25%三乙胺的20 mM磷酸钠缓冲溶液(pH 8.0)-乙腈 (60:40, v/v)。分析物通过在激发和发射波长分别为 337 和 405 nm 下使用荧光检测进行检测。该方法在 3–3000 ng/mL 范围内对所有三种分析物均呈线性，色谱运行时间为 9 分钟。日内和日间精度的值范围从 1.3 到 7.3。该方法已成功应用于定量 92 名 SLE 患者血液中 HCQ 及其两种代谢物的浓度。