SIMULTANEOUS DETERMINATION OF ARTESUNATE AND AMODIAQUINE IN HUMAN PLASMA USING LC-MS/MS AND ITS APPLICATION TO A PHARMACOKINETIC STUDY
Objective: The objective of this research was to develop a simple, rapid and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of artesunate and amodiaquine in human plasma.
Methods: An analytical method based on LC-MS/MS has been developed and validated for the simultaneous determination of artesunate and amodiaquine in human plasma. Isotope-labeled compounds are used as internal standards for the quantification of these drugs. Analytes were extracted from the plasma using solid phase extraction (SPE) technique and chromatographed on a C8 column using an isocratic mobile phase composed of 0.1% ammonia solution and methanol (10:90, v/v). The mobile phase was pumped at a flow rate of 1.00 ml/min. A total of five analytical batches were generated for the calculation of intra-day and inter-day precision and accuracy during the entire course of validation.
Results: The assay exhibits excellent linearity in the concentration range of 3.07â€“305.29 ng/ml for artesunate and 0.30â€“30.01 ng/ml for amodiaquine. Intra-day and inter-day precision and accuracy results are well within the acceptance limits. All the stability experiments were conducted in plasma samples and in neat samples are complying with the recent US FDA and EMEA guidelines.
Conclusion: The proposed LCâ€“MS/MS assay method is simple, rapid and sensitive enough for the simultaneous determination of artesunate and amodiaquine in human plasma. This method was successfully used to quantitate the in-vivo plasma concentrations obtained from a pharmacokinetic study and the results were validated by conducting incurred samples reanalysis (ISR).
2. Rosenthal PJ. Artesunate for the treatment of severe falciparum malaria. N Engl J Med 2008;358:1829-36.
3. Anvikar AR, Sharma B, Shahi BH, Tyagi PK, Bose TK, Sharma SK, et al. Artesunate-amodiaquine fixed dose combination for the treatment of Plasmodium falciparum malaria in India. Malar J 2012;11:97.
4. Koram KA, Quaye L, Abuaku B. Efficacy of Amodiaquine/ Artesunate combination therapy for uncomplicated malaria in children under five years in Ghana. Ghana Med J 2008;42:55â€“60.
5. MÃ¥rtensson A, StrÃ¶mberg J, Sisowath C, Msellem MI, Gil JP, Montgomery SM, et al. Efficacy of artesunate plus amodiaquine versus that of artemether-lumefantrine for the treatment of uncomplicated childhood plasmodium falciparum malaria in Zanzibar. Tanzania Clin Infect Dis 2005;41:1079-86.
6. Sirima SB, GansanÃ© A. Artesunate-amodiaquine for the treatment of uncomplicated malaria. Expert Opin Invest Drugs 2007;16:1079-85.
7. Sinou V, Malaika LT, Taudon N, Lwango R, Alegre SS, Bertaux L, et al. Pharmacokinetics and pharmacodynamics of a new ACT formulation: Artesunate/Amodiaquine (TRIMALACT) following oral administration in African malaria patients. Eur J Drug Metab Pharmacokinet 2009;34:133-42.
8. Li B, Zhang J, Zhou XZ, Li JY, Yang YJ, Wei XJ, et al. Determination and pharmacokinetic studies of artesunate and its metabolite in sheep plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B: Anal Technol Biomed Life Sci 2015;997:146-53.
9. Birgersson S, Ericsson T, Blank A, Hagens Cv, Ashton M, Hoffmann KJ. A high-throughput LC-MS/MS assay for quantification of artesunate and its metabolite dihydroartemisinin in human plasma and saliva. Bioanalysis 2014;6:2357-69.
10. Geditz MC, Heinkele G, Ahmed A, Kremsner PG, Kerb R, Schwab M, et al. LC-MS/MS method for the simultaneous quantification of artesunate and its metabolites dihydroartemisinin and dihydroartemisinin glucuronide in human plasma. Anal Bioanal Chem 2014;406:4299-308.
11. He G, Qi H, Wang M, Yang J, Wen F, Wang W, et al. LC-MS/MS method for the simultaneous quantitation of three active components derived from a novel prodrug against schistosome infection. J Pharm Biomed Anal 2013;83:186-93.
12. Duthaler U, Keiser J, Huwyler J. Development and validation of a liquid chromatography and ion spray tandem mass spectrometry method for the quantification of artesunate, artemether and their major metabolites dihydroartemisinin and dihydroartemisinin-glucuronide in sheep plasma. J Mass Spectrom 2011;46:172-81.
13. Thuy le TD, Hung le N, Danh PT, Na-Bangchang K. Development and validation of a liquid chromatography-mass spectrometry method for the simultaneous quantification of artesunate and dihydroartemisinin in human plasma. Southeast Asian J Trop Med Public Health 2008;39:963-77.
14. Gu Y, Li Q, Melendez V, Weina P. Comparison of HPLC with electrochemical detection and LC-MS/MS for the separation and validation of artesunate and dihydroartemisinin in animal and human plasma. J Chromatogr B: Anal Technol Biomed Life Sci 2008;867:213-8.
15. Van Quekelberghe SA, Soomro SA, Cordonnier JA, Jansen FH. Optimization of an LC-MS method for the determination of artesunate and dihydroartemisinin plasma levels using liquid-liquid extraction. J Anal Toxicol 2008;32:133-9.
16. Naik H, Murry DJ, Kirsch LE, Fleckenstein L. Development and validation of a high-performance liquid chromatography-mass spectroscopy assay for determination of artesunate and dihydroartemisinin in human plasma. J Chromatogr B: Anal Technol Biomed Life Sci 2005;816:233-42.
17. Stepniewska K, Taylor W, Sirima SB, Ouedraogo EB, Ouedraogo A, GansanÃ© A, et al. Population pharmacokinetics of artesunate and amodiaquine in African children. Malar J 2009;8:200.
18. Chen X, Deng P, Dai X, Zhong D. Simultaneous determination of amodiaquine and its active metabolite in human blood by ion-pair liquid chromatography-tandem mass spectrometry. J Chromatogr B: Anal Technol Biomed Life Sci 2007;860:18-25.
19. Lai CS, Nair NK, Muniandy A, Mansor SM, Olliaro PL, Navaratnam V. Validation of high-performance liquid chromatography-electrochemical detection methods with simultaneous extraction procedure for the determination of artesunate, dihydro-artemisinin, amodiaquine and desmethyl amodiaquine in human plasma for application in clinical pharmacological studies of the artesunate-amodiaquine drug combination. J Chromatogr B: Anal Technol Biomed Life Sci 2009;877:558-62.
20. Hodel EM, Zanolari B, Mercier T, Biollaz J, Keiser J, Olliaro P, et al. A single LC-tandem mass spectrometry method for the simultaneous determination of 14 antimalarial drugs and their metabolites in human plasma. Chromatogr B: Analyt Technol Biomed Life Sci 2009;877:867-86.
21. Katteboina MY, Pilli NR, Inamadugu JK, Satla SR. LC-MS/MS assay for irbesartan in human plasma using solid phase extraction technique: a pharmacokinetic study. Int J Pharm Pharm Sci 2015;7:335-40.
22. US DHHS, FDA and CDER. Guidance for Industry: Bioanalytical Method Validation. US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research and Center for Veterinary Medicine; 2001. Available from: http://www/fda.gov/cder/ guidance/ index.htm. [Last accessed on 02 Jul 2015].
23. Guideline on bioanalytical method validation, Science and Medicinal Health, European Medicines Agency (EMEA), EMEA/CHMP/EWP/192217/2009; 2011.
24. Viswanathan CT, Bansal S, Booth B, De Stefano AJ, Rose MJ, Sailstad J, et al. Quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. Pharm Res 2007;24:1962â€“73.
25. De Boer T, Wieling J. Incurred sample accuracy assessment: design of experiments based on standard addition. Bioanalysis 2011;3:983â€“92.
26. Raghunadha Reddy S, Naidu YK, Koteswara Rao D, Ravikiran V, Sandhya Rani Y, Kalyan Reddy MP. Development and validation of high-performance liquid chromatography-tandem mass spectrometric method for quantification of aceclofenac in human plasma. J Pharm Chem 2010;4:89â€“95.
27. Raghunadha Reddy S, Sarath Chandiran I, Jayaveera KN, Koteswara Rao D. Quantification of ibuprofen in human plasma by using high throughput liquid chromatographyâ€“tandem mass spectrometric method and its applications in pharmaco-kinetics. Arch Appl Sci Res 2010;2:101-11.