VALIDATION OF 6-MERCAPTOPURINE AND 6-THIOGUANINE IN DRIED BLOOD SPOT BY ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY
DOI:
https://doi.org/10.22159/ijap.2018.v10s1.91Keywords:
6-Mercaptopurine, 6-Thioguanine, Dried blood spot, Ultra-high-performance liquid chromatography-tandem mass spectrometryAbstract
Objective: This study aimed to obtain an optimal and validated method of analyzing 6-mercaptopurine (6-MP) and 6-thioguanine (6-TG) simultaneously
in dried blood spot samples using ultra high-performance liquid chromatography-tandem mass spectrometry.
Method: Separation was performed with a 1.7-μm amide column, which had a mobile phase with a flow rate of 0.2 mL/min and comprised 0.2%
formic acid in water, 0.1% formic acid in acetonitrile, and methanol with a gradient elution. Detection was performed using Waters Xevo TQD.
Result: This method was linear with a range of 25–1000 ng/mL for 6-MP and 6-TG, with consecutive r values of ≥0.996 and ≥0.995, respectively. The
intra- and inter-day % difference value and coefficient of variation for the accuracy and precision were not more than 15% and 20%, respectively, at
a concentration lower limit of quantitation.
Conclusion: This method fulfilled the requirements of the European Medicines Agency guideline for validation.
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