Kaewkhao Natpapat, Hanpithakpong Warunee, Tarning Joel, Blessborn Daniel
Mahidol Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand.
Centre for Tropical Medicine & Global Health, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, UK.
Wellcome Open Res. 2024 Aug 5;9:231. doi: 10.12688/wellcomeopenres.20613.2. eCollection 2024.
Ivermectin is a widely used drug for the treatment of helminthiasis and filariasis worldwide, and it has also shown promise for malaria elimination through its potent mosquito-lethal activity. The objective of this study was to develop and validate a high-throughput and sensitive method to quantify ivermectin in plasma and whole blood samples, using automated sample extraction followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Phospholipids were removed in patient whole blood (100 µl) and plasma (100 µl) samples using a 96-well plate Hybrid-solid phase extraction technique. Ivermectin and its isotope-labelled internal standard (ivermectin-D2) were separated on an Agilent Poroshell 120 EC-C18 50mm × 3.0mm I.D. 2.7µm, using a mobile phase of acetonitrile: ammonium formate 2 mM containing 0.5% formic acid (90: 10, v/v). Detection was performed using a triple quadrupole mass spectrometer in the positive ionization mode.
The method was validated in the concentration range 0.970 - 384 ng/ml in both plasma and whole blood matrices. Intra- and inter-batch precisions during the validation were below 15%. There was no carryover or matrix effects detected. Ivermectin is a stable compound and results showed no degradation in the different stability tests.
The validated method proved to have high sensitivity and precision, good selectivity and to be suitable for clinical application or laboratory quantification of ivermectin in plasma or whole blood samples.
伊维菌素是一种在全球广泛用于治疗蠕虫病和丝虫病的药物，并且通过其强大的杀蚊活性，在疟疾消除方面也显示出前景。本研究的目的是开发并验证一种高通量且灵敏的方法，用于定量血浆和全血样本中的伊维菌素，该方法采用自动样本提取，随后进行液相色谱 - 串联质谱法（LC-MS/MS）。
使用96孔板混合固相萃取技术去除患者全血（100μl）和血浆（100μl）样本中的磷脂。伊维菌素及其同位素标记的内标（伊维菌素 - D2）在安捷伦Poroshell 120 EC-C18 50mm×3.0mm内径2.7μm色谱柱上分离，流动相为乙腈：含0.5%甲酸的2 mM甲酸铵（90:10，v/v）。使用三重四极杆质谱仪在正离子模式下进行检测。
该方法在血浆和全血基质中的浓度范围为0.970 - 384 ng/ml内得到验证。验证期间的批内和批间精密度均低于15%。未检测到残留或基质效应。伊维菌素是一种稳定的化合物，结果表明在不同的稳定性试验中没有降解。
验证后的方法具有高灵敏度和精密度、良好的选择性，适用于临床应用或实验室对血浆或全血样本中伊维菌素的定量。
