Qin Chaolong, Feng Wanshan, Chu YenJu, Lee Jong Bong, Berton Mattia, Bettonte Sara, Teo Yeong Yeu, Stocks Michael J, Fischer Peter M, Gershkovich Pavel
School of Pharmacy, University of Nottingham, Nottingham, UK.
Department of Pharmaceutical and Pharmacological Science, University of Padova, Padova, Italy.
Biomed Chromatogr. 2020 Nov;34(11):e4934. doi: 10.1002/bmc.4934. Epub 2020 Jul 10.
A simple, sensitive and cost-effective HPLC-UV bioanalytical method for determination of lopinavir (LPV) in rat and human plasma was developed and validated. The plasma sample preparation procedure includes a combination of protein precipitation using cold acetonitrile and liquid-liquid extraction with n-hexane-ethyl acetate (7:3, v/v). A good chromatographic separation was achieved with a Phenomenex Gemini column (C , 150 mm × 2.0 mm, 5 μm) at 40°C with gradient elution, at 211 nm. Calibration curves were linear in the range 10-10,000 ng/mL, with a lower limit of quantification of 10 ng/mL using 100 μL of plasma. The accuracy and precision in all validation experiments were within the criteria range set by the guidelines of the Food and Drug Administration. This method was successfully applied to a preliminary pharmacokinetic study in rats following an intravenous bolus administration of LPV. Moreover, the method was subsequently fully validated for human plasma, allowing its use in therapeutic drug monitoring (TDM). In conclusion, this novel, simple and cost-efficient bioanalytical method for determination of LPV is useful for pharmacokinetic and drug delivery studies in rats, as well as TDM in human patients.
建立并验证了一种简单、灵敏且经济高效的HPLC-UV生物分析方法，用于测定大鼠和人血浆中的洛匹那韦（LPV）。血浆样品制备程序包括使用冷乙腈进行蛋白沉淀以及用正己烷-乙酸乙酯（7:3，v/v）进行液-液萃取。使用Phenomenex Gemini柱（C ，150 mm × 2.0 mm，5 μm）在40°C下通过梯度洗脱在211 nm处实现了良好的色谱分离。校准曲线在10 - 10,000 ng/mL范围内呈线性，使用100 μL血浆时定量下限为10 ng/mL。所有验证实验中的准确度和精密度均在食品药品监督管理局指南设定的标准范围内。该方法成功应用于大鼠静脉推注LPV后的初步药代动力学研究。此外，该方法随后在人血浆中进行了全面验证，可用于治疗药物监测（TDM）。总之，这种用于测定LPV的新颖、简单且经济高效的生物分析方法可用于大鼠的药代动力学和药物递送研究以及人类患者的TDM。
