Development, validation, and clinical application of a rapid UPLC–MS/MS method for detection of colchicine in human whole blood and urine

Abstract

A rapid, simple, and economical method has been developed to determine colchicine in both human whole blood and urine using UPLC-MS/MS. Colchicine and isotopelabeled internal standard were extracted from the matrix by liquid-liquid extraction with saturated borax and ethyl acetate, and separated by a reversed-phase chromatography C18 column. Gradient elution was carried out using acetonitrile and water spiked with 0.01% formic acid. Multiple reaction monitoring was performed at positive ion mode. The quantitative transitions were m/z 400.27 ! 310.28 for colchicine and m/z 406.16 ! 313.18 for colchicine-D6. The method has good linearity in the range of 0.5-200 ng/mL for blood and 2-2000 ng/mL for urine. The sensitivity, accuracy, and matrix effect were all in line with the guidelines of Food and Drug Administration and European Medicines Agency. The extraction recovery was above 63.94%. The samples were stable under various storage conditions. Six deuteriumsubstituted isotopic internal standard was used to demonstrate a different mode of colchicine cleavage from the existing literature. This method has been successfully used in the diagnosis of patients with colchicine poisoning. Blood is recommended as the optimal sample compared with urine.