Žideková Nela, Pršo Kristián, Pršo Marek, Jeseňák Miloš, Farsa Oldřich, Kertys Martin
Biomedical Center Martin, Jessenius Faculty of Medicine in Martin, Comenius University Bratislava, Martin, Slovak Republic.
Department of Research and Development, Saneca Pharmaceuticals, Hlohovec, Slovak Republic.
Biomed Chromatogr. 2025 Nov;39(11):e70222. doi: 10.1002/bmc.70222.
Colchicine is a naturally occurring alkaloid primarily derived from plants of the Colchicum genus, which is used to treat gout and serve as a frontline therapy for various inflammatory conditions, including familial Mediterranean fever. Although it is not recommended for routine therapeutic drug monitoring, there are situations where it may be beneficial, such as in dose adjustments. The present study introduces an LC-MS/MS method for quantifying colchicine in human plasma. A one-step extraction procedure employing an Ostro plate was applied, and the extracts were analyzed using gradient elution followed by detection on a mass spectrometer in multiple reaction monitoring mode. Our method offers several advantages, including a low sample volume and a run time of only 3 min. It demonstrates sufficient linearity to quantify low and high concentrations of colchicine in human plasma samples. The method was successfully validated in accordance with the ICH guideline M10 on bioanalytical method validation, covering selectivity, linearity, limit of quantification, accuracy, precision, dilution integrity, carry-over effect, matrix effects, extraction recovery, and stability over a concentration range of 0.05-100 ng/mL. The fully developed and validated method was applied to determine colchicine in plasma samples from patients diagnosed with familial Mediterranean fever.
秋水仙碱是一种天然存在的生物碱，主要从秋水仙属植物中提取，用于治疗痛风，并作为包括家族性地中海热在内的各种炎症性疾病的一线治疗药物。虽然不建议进行常规治疗药物监测，但在某些情况下可能有益，例如在剂量调整时。本研究介绍了一种用于定量人血浆中秋水仙碱的液相色谱-串联质谱法。采用了使用Ostro板的一步提取程序，并使用梯度洗脱对提取物进行分析，然后在多反应监测模式下在质谱仪上进行检测。我们的方法具有几个优点，包括低样品体积和仅3分钟的运行时间。它显示出足够的线性度，可对人血浆样品中低浓度和高浓度的秋水仙碱进行定量。该方法已按照国际人用药品注册技术协调会（ICH）关于生物分析方法验证的M10指南成功验证，涵盖了选择性、线性度、定量限、准确度、精密度、稀释完整性、残留效应、基质效应、提取回收率以及在0.05-100 ng/mL浓度范围内的稳定性。该完全开发和验证的方法用于测定诊断为家族性地中海热的患者血浆样品中的秋水仙碱。
