THE EFFECT OF ANTICOAGULANT TYPES ON THE IN VITRO ANALYSIS OF CLOPIDOGREL IN HUMAN PLASMA USING LIQUID CHROMATOGRAPHY TANDEM-MASS SPECTROMETRY

Authors

  • Yahdiana Harahap Faculty of Pharmacy, Universitas Indonesia, Depok 16424, Indonesia
  • Anisa Maulidina Faculty of Pharmacy, Universitas Indonesia, Depok 16424, Indonesia
  • Delly Ramadon Faculty of Pharmacy, Universitas Indonesia, Depok 16424, Indonesia

DOI:

https://doi.org/10.22159/ijap.2018.v10s1.87

Keywords:

Citrate, Clopidogrel, Ethylenediaminetetraacetic acid, Heparin, Liquid chromatography tandem-mass spectrometry

Abstract

Objective: The aim of this study was to optimize and validate a plasma clopidogrel analysis method using liquid chromatography tandem-mass
spectrometry.
Methods: Plasma samples were analyzed using a BEH C18 column (1.7 μm; 100 mm×2.1 mm), the mobile phase was 0.1% formic acid in acetonitrile
(30:70, v/v). The flow rate was 0.2 mL/min, with a column temperature set to 35°C, an injection volume of 5 μL, an analysis time of 4 min, and
irbesartan as the internal standard. Aliquots were obtained by liquid-liquid extraction using ammonium acetate and diethyl ether. The stability and
peak area ratio of the respective plasma area responses were evaluated using ANOVA.
Results: No significant differences (p>0.05) were observed between anticoagulants regarding analyte stability. However, the peak area ratio
showed significant differences (p<0.05) between the anticoagulants. The accuracy and precision of the analysis with citrate, heparin, and
ethylenediaminetetraacetic acid (EDTA) plasma met the quality requirements, and a linear calibration curve was created with concentrations ranging
from 0.02 to 5.0 ng/mL.
Conclusion: The results showed that improved analysis of clopidogrel was achieved using citrate or heparin plasma compared with EDTA plasma.

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References

Rodak BF, Fritsma GA, Keohane EM. Hematology Clinical Principles

and Applications. Hematology Clinical Principles and Applications.

Missouri: Elseviers; 2012.

Wells BG, DiPiro JT, Schwinghammer TL, DiPiro CV. Pharmacotherapy

Handbook. 9th ed. New York: McGraw-Hill; 2015.

Lüllmann H, Mohr K, Hein L, Bieger D. Color Atlas of Pharmacology.

rd ed. New York: Georg Thieme Verlag; 2005.

Lainesse A, Ozalp Y, Wong H, Alpan RS. Bioequivalence study of

clopidogrel bisulfate film-coated tablets. Arzneimittel Forschung

;54:600-4.

Bowen RA, Hortin GL, Csako G, Otañez OH, Remaley AT. Impact of

blood collection devices on clinical chemistry assays. Clin Biochem

;43:4-25.

Barri T, Dragsted LO. UPLC-ESI-QTOF/MS and multivariate data

analysis for blood plasma and serum metabolomics: Effect of experimental

artefacts and anticoagulant. Anal Chim Acta 2013;768:118-28.

Li W, Zhang J, Tse FL, Li W, Zhang J, Tse FL, editors. Handbook of

LC-MS Bioanalysis : Best Practices, Experimental Protocols, and

Regulations. New Jersey: John Wiley and Sons; 2013.

Moffat AC, Osselton M, Widdop B, Moffat AC, Osselton M, Widdop B,

editors. Clark’s Analysis of Drugs and Poisons. 3rd ed. London:

Pharmaceutical Press; 2004.

Christensen JM, Stalker D. Ibuprofen piconol hydrolysis in vitro in

plasma, whole blood, and serum using different anticoagulants. J Pharm

Sci 1991;80:29-31.

Barton RH, Waterman D, Bonner FW, Holmes E, Clarke R, Procardis

Consortium, et al. The influence of EDTA and citrate anticoagulant

addition to human plasma on information recovery from NMR-based

metabolic profiling studies. Mol Biosyst 2010;6:215-24.

Evans G. A Handbook of Bioanalysis and Drug Metabolism.

A Handbook of Bioanalysis and Drug Metabolism. Boca Raton: CRC

Press; 2004.

Yi J, Craft D, Gelfand CA. Minimizing preanalytical variation of

plasma samples by proper blood collection and handling. Methods Mol

Biol 2011;728:137-49.

Gonzalez-Covarrubias V, Dane A, Hankemeier T, Vreeken RJ. The

influence of citrate, EDTA, and heparin anticoagulants to human

plasma LC–MS lipidomic profiling. Metabolomics 2013;9:337-48.

Harahap Y. The Role of Bioanalysis in Drug Quality Assurance and

Improving Quality of Patient Life. Jakarta: Universitas Indonesia-

Press; 2010.

Hall TG, Smukste I, Bresciano KR, Wang Y, Mckearn D, Savage RE.

Identifying and overcoming matrix effects in drug discovery and

development. In: Tandem Mass Spectrometry-Applications and

Principles. INTECH; 2012. p. 794.

Harahap Y, Masyarah I, Suryadi H. Analytical validation of clopidogrel in

human plasma through ultra-high performance liquid chromatographytandem

mass spectrometry. Int J App Pharm 2017;9:721-33.

Waters. Waters UPLC, UHPLC, and HPLC Column Selection and

Mobile Phase Guide Wide pH Range. USA: Waters Corporation; 2016.

Shaikh S, Jain V. A novel reverse-phase high-performance liquid

chromatographic method for simultaneous estimation of ellagic acid,

quercetin, and piperine in ayurvedic formulations. Asian J Pharm Clin

Res 2018;11:312-7.

Rao N, Gawde K. Method development and force degradation

studies for simultaneous estimation of salbutamol sulfate, etofylline

and bromhexine hydrochloride in pharmaceutical dosage form using

reversed-phase high-performance liquid chromatography method.

Asian J Pharm Clin Res 2018;11:378-82.

Selvaraj NA, Vijayalkshmi K. Chromatographic separation of bioactive

compounds from ipomoeae batatas lam by Column, HPTLC and GCMS

techniques. Asian J Pharm Clin Res 2014;7:4-8.

Fountain KJ. UPLC Versus UHPLC : Comparison of Loading and

Peak Capacity for Small Molecule Drugs. Milford, USA: Waters

Corporation; 2011. p. 1-6.

European Medicines Agency. Guideline on the Investigation of

Bioequivalence. Vol 1. London: European Medicines Agency; 2010.

Published

20-12-2018

How to Cite

Harahap, Y., Maulidina, A., & Ramadon, D. (2018). THE EFFECT OF ANTICOAGULANT TYPES ON THE IN VITRO ANALYSIS OF CLOPIDOGREL IN HUMAN PLASMA USING LIQUID CHROMATOGRAPHY TANDEM-MASS SPECTROMETRY. International Journal of Applied Pharmaceutics, 10(1), 392–395. https://doi.org/10.22159/ijap.2018.v10s1.87

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