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

Abstract


 

 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-CAMAG® 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 Acquity® 1.7 μm (2.1 mm × 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/8×108 erythrocytes, respectively. The levels of 6-MMP gained 16.59 pmol/8×108 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.


Keywords


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

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About this article

Title

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

Topics

Pharmacy,

Keywords

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

DOI

10.22159/ajpcr.2017.v10i9.19790

Date

01-09-2017

Additional Links

Manuscript Submission

Journal

Asian Journal of Pharmaceutical and Clinical Research
Vol 10 Issue 9 September 2017 Page: 120-125

Print ISSN

0974-2441

Online ISSN

2455-3891

Authors & Affiliations

Supandi Supandi
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia
Indonesia

Yahdiana Harahap
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia
Indonesia

Harmita Harmita
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia
Indonesia

Rizka Andalusia
Department of Research and Development, Dharmais Cancer Hospital, Jakarta 11420, Indonesia.
Indonesia

Marlina Ika
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok Campus 16424, Indonesia
Indonesia


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