SIMULTANEOUS ANALYTICAL METHOD DEVELOPMENT OF 6-MERCAPTOPURINE AND 6-METHYLMERCAPTOPURINE IN DRIED BLOOD SPOT USING ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY


Marlina Ika, Rizka Andalusia, Supandi Supandi, Yahdiana Harahap

Abstract


Objective: 6-mercaptopurine (6-MP) is a chemotherapeutic agent in the antimetabolite class. It has to go through the metabolic pathway to form
6-methyl MP (6-MMP). This study aimed to obtain an optimum and validated method for the analysis of 6-MP and 6-MMP in dried blood spot (DBS)
samples simultaneously and to evaluate the potential for future drug concentration monitoring in DBS samples.
Methods: The quality control and calibration curves were made by spotting 40 μL blood on DBS paper and dried for 3 hrs. DBS papers were cut with a
diameter of 8 mm and extracted with acetonitrile-methanol (1:3) containing internal standard 5-fluorouracil (5-FU). Separation was performed with
waters acquity ultra performance liquid chromatography BEH C18 column of 1.7 μm (2.1×100 mm) with a mobile phase consisting of 0.1% formic
acid in water 0.1% formic acid in acetonitrile with gradient elution and a flow rate of 0.2 mL/minute. Mass detection was performed using Waters
Xevo TQD with positive electrospray ionization (ESI) for 6-MP and 6-MMP and negative ESI for 5-FU in the multiple reaction monitoring mode.
Results: The detection rates of 6-MP, 6-MMP, and 5-FU were 153.09>119.09, 167.17>126.03, and 129.09>42.05, respectively. This method was linear
with the range at 26-1000 ng/mL for 6-MP and 13-500 ng/mL for 6-MMP with consecutive r≥0.998 and ≥0.999, respectively. The % relative error
value and % relative standard deviation for accuracy and precision of intraday and interday were not more than 15% and not more than 20% at the
lower limit of quantification concentration, respectively.
Conclusions: This method fulfilled the requirements of selectivity, linearity, carry over, and matrix effects referring to the European Medicines Agency
guidelines.


Keywords


6-Mercaptopurine, 6-Methylmercaptopurine, Dried blood spot, Ultra performance liquid chromatography tandem mass spectrometry, Validation.

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About this article

Title

SIMULTANEOUS ANALYTICAL METHOD DEVELOPMENT OF 6-MERCAPTOPURINE AND 6-METHYLMERCAPTOPURINE IN DRIED BLOOD SPOT USING ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY

Keywords

6-Mercaptopurine, 6-Methylmercaptopurine, Dried blood spot, Ultra performance liquid chromatography tandem mass spectrometry, Validation.

DOI

10.22159/ijap.2017.v9s1.80_87

Date

30-10-2017

Additional Links

Manuscript Submission

Journal

International Journal of Applied Pharmaceutics
Vol. 9, Special Issue (Oct.), 2017 [PTMDS 2017] Page: 168-171

Online ISSN

0975-7058

Statistics

7 Views | 21 Downloads

Authors & Affiliations

Marlina Ika
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
Indonesia

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

Supandi Supandi
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
Indonesia

Yahdiana Harahap
Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
Indonesia


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