ANALYSIS OF 6-MERCAPTOPURINE AND 6-METHYLMERCAPTOPURINE IN DRIED BLOOD SPOTS USING LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY AND ITS APPLICATION IN CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA PATIENTS

Supandi Supandi; Yahdiana Harahap; Harmita Harmita; Rizka Andalusia; Marlina Ika

doi:10.22159/ajpcr.2017.v10i9.19790

Authors

Supandi Supandi Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia http://orcid.org/0000-0002-0063-2548
Yahdiana Harahap Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia
Harmita Harmita Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia
Rizka Andalusia Department of Research and Development, Dharmais Cancer Hospital, Jakarta 11420, Indonesia.
Marlina Ika Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia

DOI:

https://doi.org/10.22159/ajpcr.2017.v10i9.19790

Keywords:

6-mercaptopurine, 6-methylmercaptopurine, Dried blood spots, Acute lymphoblastic leukemia

Abstract

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Â Objective: To analyze and validate 6-mercaptopurine (6-MP) and 6-methylmercaptopurine (6-MMP) in dried blood spots (DBS) using liquid chromatography-tandem mass spectrometry (LC/MS-MS).

Methods: Bio-sampling dried blood spot with DBS-CAMAGÂ® paper diameter of 8 mm and extracted with acetonitrile-methanol (1:3) containing internal standard 5-fluorouracil (5-FU). Separation was performed with C18 column AcquityÂ® 1.7 Î¼m (2.1 mm Ã— 100 mm), with a mobile phase mixture of 0.1% formic acid in water 0.1% formic acid in acetonitrile with gradient elution and flow rate 0.2 ml/min. Mass detection was Waters Xevo TQD with positive electrospray ionization (ESI) for 6-MP, 6-MMP and negative ESI for 5-FU in multiple reaction monitoring modes. The ions of 6-MP was detected at m/z 153.09->119.09, 6-MMP at m/z 167.17->126.03, and 5-FU at m/z 129.15->42.05.

Results: This method fulfill the requirements of selectivity, linearity, lower limit of quantification, accuracy, precision, carry-over, matrix effects, and stability which refers to the european medicines agency (EMEA) guidelines. The linearity of 0.99 was 26-1000 ng/mL for 6-MP and 6-MMP, respectively. The validated method was applied to two childhood ALL maintenance phase. Retrieved 6-MP levels of 10.37 pmol/8Ã—108 erythrocytes, respectively. The levels of 6-MMP gained 16.59 pmol/8Ã—108 erythrocytes, respectively.

Conclusion: The developed LC/MS-MS method is valid to analysis 6-MP and 6-MMP in DBS simultaneous in vitro according to EMEA guidelines. The method was successfully applied to authentic capillary blood samples from two childhood patients with ALL in the maintenance phase.

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