Development and Validation of an LC-MS/MS Method for the Determination of Lamivudine, Tenofovir & Efavirenz in human plasma
DOI:
https://doi.org/10.63682/jns.v14i19S.4966Keywords:
Lamivudine, Tenofovir, Efavirenz, Abacavir and EmtricitabineAbstract
Introduction: Therapeutic drug monitoring of Lamivudine, tenofovir and efavirenz, three commonly used antiretroviral drugs, is important to maximize effectiveness while minimizing side effects.
Materials and Methods:
Chromatographic separation was done on Xterra, C18 (2), 150 X 4.6 mm, 5 µm column with a mobile phase composed of 10mM ammonium formate-methanol-acetonitrile in the ratio of 50:25:25 (v/v/v), at a flow rate of 1.0 mL/min. Turbo ion-spray interface (TIS) operated in positive ionization mode was used for the mass spectrometric detection. The MRM transitions monitored were m/z 230.1/112.1
(lamivudine), m/z 288.0/176.2 (tenofovir), m/z 316.2/168.1 (Efavirenz), m/z 248.1/130.1 (Emtricitabine,ISTD) and m/z 287.2/191.2(Abacavir,ISTD). The method was developed and the established calibration ranges are 10-3018 ng/mL, 5-500ng/mL and 20-6021ng/mLfor lamivudine, tenofovir disoproxilfumarateand efavirenz respectively. The slope values are consistent and regression values were found to be more than or equal to 0.99. The accuracy for run size reproducibility batch and dilution integrity (1/2, 1/4) for lamivudine, tenofovir disoproxilfumarateand efavirenz was found to be in the range of 96.9-102.2%. The precision was found to be less than 8.1% for three analytes. Further, the reported method was validated as per the ICH guidelines and found to be well with in the acceptable range.
Results: The method showed good accuracy, low limits of quantification, adequate recovery, minimal matrix effects, and specificity. Analyte stability under multiple storage conditions was demonstrated.
Conclusion: The validated LC-MS/MS method provides a reliable tool for therapeutic drug monitoring and pharmacokinetic studies of anti-HIV regimens. The assay can be applied to large populations, especially in resource-poor settings, to help individualize dosing and improve clinical outcomes while reducing toxicity
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Kauhanen D, Sysi-Aho M, Koistinen KM, Laaksonen R, Sinisalo J, Ekroos K. Development and validation of a high-throughput LC–MS/MS assay for routine measurement of molecular ceramides. Anal Bioanal Chem. 2016;408:3475-3483. doi:10.1007/s00216-016-9425-z
Raćkowska E, Bobrowska-Korczak B, Giebułtowicz J. Development and validation of a rapid LC–MS/MS method for determination of methylated nucleosides and nucleobases in urine. J Chromatogr B. 2019;1128:121775. doi:10.1016/j.jchromb.2019.121775
Lucatello L, Cagnardi P, Capolongo F, Ferraresi C, Bernardi F, Montesissa C. Development and validation of an LC–MS/MS/MS method for the quantification of fluoroquinolones in several matrices from treated turkeys. Food Control. 2015;48:2-11. doi:10.1016/j.foodcont.2014.04.011
Onmaz DE, Abusoglu S, Onmaz M, Yerlikaya FH, Unlu A. Development and validation of a sensitive, fast and simple LC-MS/MS method for the quantitation of favipiravir in human serum. J Chromatogr B. 2021;1176:122768.
Kromdijk W, Pereira S, Rosing H, Mulder JW, Beijnen JH, Huitema ADR. Development and validation of an assay for the simultaneous determination of zidovudine, abacavir, emtricitabine, lamivudine, tenofovir and ribavirin in human plasma using liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2013;919-920:43-51. doi:10.1016/j.jchromb.2013.01.005
Paliwal N. A liquid chromatography tandem mass spectrometry based regulatory compliant method for the determination of tenofovir in human serum. Drug Res. Published online 2013:306-312.
Beer B. Development and validation of a liquid chromatography–tandem mass spectrometry method for the simultaneous quantification of tamoxifen, anastrozole, and letrozole in human plasma and its application to a clinical study. Anal Bioanal Chem. 2010;398:1791-1800. doi:10.1007/s00216-010-4075-z
Mistri HN, Jangid AG, Pudage A, Shrivastav P. HPLC–ESI-MS/MS validated method for simultaneous quantification of zopiclone and its metabolites, N-desmethyl zopiclone and zopiclone-N-oxide in human plasma. J Chromatogr B. Published online 2008. doi:10.1016/j.jchromb.2008.02.004
Goswami D. Liquid Chromatographic tandem mass spectrometric validated method for pharmacokinetic estimation of flecainide in human plasma. Clin Res Regul Aff. 2009;26(1-2):24-36. doi:10.1080/10601330902919797
Wang X, Qi W, Zhao X, et al. Determination of four phenolic endocrine disruptors in environmental water samples by high performance liquid chromatography-fluorescence detection using dispersive liquid-liquid microextraction coupled with derivatization. Se Pu Chin J Chromatogr. 2014;32(6):623-628. doi:10.3724/SP.J.1123.2014.02012
Zeng M. An automated dual-gradient liquid chromatography–MS/MS method for the simultaneous determination of ferulic acid, ligustrazine and ligustilide in rat plasma and its application to a pharmacokinetic study. J Pharm Biomed Anal. 2014;88:354-363. doi:10.1016/j.jpba.2013.08.038
Girault J, Fourtillan JB. Quantitative measurement of clonidine in human plasma by combined gas chromatography/electron capture negative ion chemical ionization mass spectrometry. Biomed Env Mass Spectrom. 1988;17:443-448. doi:10.1002/bms.1200170605
Christianson CD. Development and validation of an HPLC–MS/MS method for the analysis of dexamethasone from pig synovial fluid using dried matrix spotting. Bioanalysis. 2010;2(11):1829-1837. doi:10.4155/bio.10.137
Jain DS. A high throughput and selective method for the estimation of valproic acid an antiepileptic drug in human plasma by tandem LC–MS/MS. Talanta. 2007;72(1):80-88. doi:10.1016/j.talanta.2006.09.033
Patel DP. UPLC‐MS/MS assay for the simultaneous quantification of carvedilol and its active metabolite 4′‐hydroxyphenyl carvedilol in human plasma to support a bioequivalence study in healthy volunteers. Biomed Chromatogr. 2013;27(8):974-986.
Nagaraju P, Kodali B, Datla PV, Kovvasu SP. LC-MS/MS Quantification of Tramadol and Gabapentin Utilizing Solid Phase Extraction". Int J Anal Chem. 2018;2018, Article ID 1605950:9 ,. doi:10.1155/2018/1605950
Huang MQ. UPLC–MS/MS assay for the simultaneous determination of ethinyl estradiol, norgestimate and 17-Desacetyl norgestimate at low pg/mL in human plasma. J Chromatogr B. 2016;1017:1-9. doi:10.1016/j.jchromb.2016.01.054
Kuhn J, Knabbe C. Fully validated method for rapid and simultaneous measurement of six antiepileptic drugs in serum and plasma using ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry. Talanta. 2013;110:71-80. doi:10.1016/j.talanta.2013.02.010
Larsson M. Determination of ximelagatran, an oral direct thrombin inhibitor, its active metabolite melagatran, and the intermediate metabolites, in biological samples by liquid chromatography-mass spectrometry". J Chromatogr B. Published online 2003. doi:10.1016/S1570-0232(02)00768-7
Mazzarino M, Torre X, Botrè F. Comparison between reverse phase and hydrophilic interaction liquid chromatography techniques in doping analysis: a case study on phenolalkylamines. InRecent Adv Doping Anal. 2011;19.
Vancea S. Rapid LC–MS/MS method for determination of drotaverine in a bioequivalence study. J Pharm Biomed Anal. 2014;98:417-423. doi:10.1016/j.jpba.2014.06.029
Cheng C, Liu S, Xiao D, et al. LC–MS/MS method development and validation for the determination of polymyxins and vancomycin in rat plasma. J Chromatogr B. 2010;878(28):2831-2838.
Zhu Z, Neirinck L. High-performance liquid chromatographic method for the determination of gabapentin in human plasma. J Chromatogr B. Published online November 5, 2002.
Rago B, Fu C. Development of a high-throughput ultra performance liquid chromatography–mass spectrometry assay to profile 18 eicosanoids as exploratory biomarkers for atherosclerotic diseases. J Chromatogr B. 2013;936:25-32. doi:10.1016/j.jchromb.2013.08.001
Pappula N, Kodali B, Datla PV. Selective and rapid determination of tadalafil and finasteride using solid phase extraction by high performance liquid chromatography and tandem mass spectrometry. J Pharm Biomed Anal. Published online 2018. doi:10.1016/j.jpba.2018.01.020
González‐Hernández I. A simple LC‐MS/MS method to determine plasma and cerebrospinal fluid levels of albendazole metabolites (albendazole sulfoxide and albendazole sulfone) in patients with neurocysticercosis. Biomed Chromatogr. 2012;26(2):267-272. doi:10.1002/bmc.1659
Samanidou VF, Nikolaidou KI, Papadoyannis IN. Development and validation of an HPLC confirmatory method for the determination of seven tetracycline antibiotics residues in milk according to the European Union Decision 2002/657/EC. J Sep Sci. 2007;30(15):2430-2439. doi:10.1002/jssc.200700057
Wei F. Simultaneous determination of 19 mycotoxins in lotus seed using a multimycotoxin UFLC-MS/MS method. J Pharm Pharmacol. 2019;71(7):1172-1183. doi:10.1111/jphp.13101
Derangula VR. Liquid chromatography–tandem mass spectrometric assay for the determination of tetrabenazine and its active metabolites in human plasma: a pharmacokinetic study. Biomed Chromatogr. 2013;27(6):792-801. doi:10.1002/bmc.2862
Zhong D. Determination of three major components of bitespiramycin and their major active metabolites in rat plasma by liquid chromatography-ion trap mass spectrometry. J Chromatogr B. 2003;791(1-2):45-53. doi:10.1016/S1570-0232(03)00205-8
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