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

  • Marlina Ika Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
  • Rizka Andalusia Department of Research and Development, Dharmais Cancer Hospital, Jakarta, Indonesia.
  • Supandi Supandi Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
  • Yahdiana Harahap Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.

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.

References

1. Lennard L. The clinical pharmacology of 6-mercaptopurine. Eur J Clin
Pharmacol 1992;43(4):329-9.
2. Beaumais TA, Fakhoury M, Medard Y, Azougagh S, Zhang D,
Yakouben K, et al. Determinants of mercaptopurine toxicity in
paediatric acute lymphoblastic leukemia maintenance therapy. Br J Clin
Pharmacol 2010;71(4):575-84.
3. Schmiegelow K, Nielsen SN, Frandsen TL, Nersting J. Mercaptopurine/
Methotrexate maintenance therapy of childhood acute lymphoblastic
leukemia: Clinical facts and fiction. J Pediatr Hematol Oncol
2014;36(7):503-17.
4. Schmiegelow K, Bretton-Meyer U 6-mercaptopurine dosage and
pharmacokinetics influence the degree of bone marrow toxicity
following high-dose methotrexate in children with acute lymphoblastic
leukemia. Leukemia 2001;15(1):74-9.
5. Kirchherr H, Shipkova M, von Ahsen N. Improved method for
therapeutic drug monitoring of 6-thioguanine nucleotides and
6-methylmercaptopurine in whole-blood by LC/MSMS using isotopelabeled
internal standards. Ther Drug Monit 2013;35(3):313-21.
6. Déglon J, Thomas A, Mangin P, Staub C. Direct analysis of dried
blood spots coupled with mass spectrometry: Concepts and biomedical
applications. Anal Bioanal Chem 2012;402(8):2485-98.
7. Wilhelm AJ, den Burger JC, Swart EL. Therapeutic drug monitoring
by dried blood spot: Progress to date and future directions. Clin
Pharmacokinet 2014;53(11):961-73.
8. Evans C, Arnold M, Bryan P, Duggan J, James CA, Li W, et al.
Implementing dried blood spot sampling for clinical pharmacokinetic
determinations: Considerations from the IQ consortium microsampling
working group. AAPS J 2015;17(2):292-300.
9. Hawwa AF, Al Bawab A, Rooney M, Wedderburn LR, Beresford MW,
McElnay JC. A novel dried blood spot-LCMS method for the
quantification of methotrexate polyglutamates as a potential marker for
methotrexate use in children. PLOS One 2014;9(2):e89908.
Statistics
188 Views | 192 Downloads
How to Cite
Ika, M., Andalusia, R., Supandi, S., & Harahap, Y. (2017). SIMULTANEOUS ANALYTICAL METHOD DEVELOPMENT OF 6-MERCAPTOPURINE AND 6-METHYLMERCAPTOPURINE IN DRIED BLOOD SPOT USING ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY. International Journal of Applied Pharmaceutics, 9, 168-171. https://doi.org/10.22159/ijap.2017.v9s1.80_87
Section
Original Article(s)