A RAPID ISOCRATIC HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC-UV) METHOD FOR THE QUANTIFICATION OF RITONAVIR IN HUMAN PLASMA

Authors

  • Hemanth Kumar A. K. National Institute for Research in Tuberculosis (ICMR), Chennai, India
  • Sudha V.
  • Leelavathi A.
  • Geetha Ramachandran

Abstract

Objective: An accurate, simple, and rapid HPLC-UV based method for the quantitative determination of ritonavir (RTV) in human plasma is developed.

Methods: The method involved deproteinization of the sample with 0.125N NaOH and methyl tertiary butyl ether and evaporated to dryness. The residue was reconstituted with mobile phase (20 mM Sodium Acetate and Acetonitrile–55:45 v/v). The analysis was done using C8 column (250 x 4.6 mm ID) and detection at a wavelength of 212 nm

Results: The method range was linear over the range 0.5–10.0 µg/ml as derived using calibration curve method. Mean intra-and inter-day variations over the ranges of the standard curves were less than 10 % and mean extraction recoveries from human plasma ranged from 96 to 110 %.

Conclusion: A rapid and accurate method for quantitation of RTV in plasma was validated. The assay spans the concentration range of clinical interest. The easy sample preparation and small sample size makes this assay highly suitable for pharmacokinetic studies of RTV in HIV-infected patients with TB.

Keywords: Ritonavir, Pharmacokinetics, HPLC, Plasma extraction, HIV

Downloads

Download data is not yet available.

References

Hammer SM, Eron JJ Jr, Reiss P, Schooley RT, Thompson MA, Walmsley S, et al. Antiretroviral treatment of adult HIV infection recommendations of the international AIDS society–USA panel. Jama 2008;300:555-70.

Thompson MA, Aberg JA, Cahn P. Antirétroviral treatment of adult HIV infection: 2010 recommendations of the International AIDS Society–USA panel. Jama 2010;304:321–33.

Janoly A, Bleyzac N, Favetta P, Gagneu MC, Bourhis Y, Coudray S, et al. Simple and rapid high-performance liquid chromatographic method for nelfinavir, M8 nelfinavir metabolite, ritonavir and saquinavir assay in plasma. J Chromatogr B 2002;780:155-60.

Walson PD, Cox S, Utkin, Gerber N, Crim L, Brady M, et al. Clinical use of a simultaneous HPLC assay for indinavir, saquinavir, ritonavir and nelfinavir in china and adults. Ther Drug Monit 2003;5:588-92.

Turner ML, Reed-Walker K, King JR, Acosta EP. Simultaneous determination of nine antiretroviral compounds in human plasma using liquid chromatography. J Chromatogr B 2003;784:331-41.

Kou, HuiJuan, Min Ye, Qiang Fu, Yang Han, XiaoLi Du, et al. Simultaneous quantification of lopinavir and ritonavir in human plasma by high performance liquid chromatography coupled with UV detection. Sci Chin Life Sci 2012;55:321-7.

Hirano, Atsushi, Masaaki Takahashi, Eri Kinoshita, Masaaki Shibata. High performance liquid chromatography using UV detection for the simultaneous quantification of the new non-nucleoside reverse transcriptase inhibitor etravirine (TMC-125), and 4 protease inhibitors in human plasma. Biol Pharm Bull 2010;33:1426-9.

Laure Elens, Sophie Veriter, Vincent Di Fazio, Roger Vanbist, Daniel, Boesmans, et al. Quantification of 8 HIV-protease inhibitors and 2 non-nucleoside reverse transcription liquid chromatography with diode array detection. Clin Chem 2009;55:170-4.

Masskai Takahashi, Masao Yoshida, Tsuyoshi Oki, Naoya Okumura, Tatsuo Suzuki and Tsuguhiro Kaneda. Conventional HPLC Method used for Simultaneous Determination of the seven HIV protease inhibitors and nonnucleoside reverse transcription inhibitors efavirenz in human plasma. Biol Pharm Bull 2005;28:1286-90.

Hirbayashi Y, Tsuchiya K, Kimura S, Oka s. Simultaneous determination of six HIV protease inhibitors (amprenavir, indinavir, lopinavir, ritonavir and squinavir), the active metabolism of nelfinavir (M8) and non-nucleoside reverse transcriptase inhibitor (efavirenz) in human plasma by high-performance liquid chromatography. Biomed Chromatogr 2006;20:28-36.

Tribut O, Verider Mc, Arvieux C, Allian H, Michelet C, Bentue–Ferrer D. Simultaneous quantitative assay of atazanavir and 6 other HIV protease inhibitors by isocratic reversed–phase liquid chromatography in human plasma. Ther Drug Monit 2005;27:265-9.

Verbesselt R, Van Wijngaerden E, De Hoon J. Simultaneous determination of 8 HIV protease inhibitors in human plasma by isocratic high-performance liquid chromatography wtih combined use of UV and fluorescence detection: amprenavir, indinavir, atazanavir, retonavir, saquinavir, nelfinavir and M8-nelfinavir metobolite. J Chromatogr B Analyt Technol Biomed Life Sci 2007;845:51-60.

Poirier JM, Robidou P, Jaillon P. Simple and simultaneous determination of the HIV–protease inhibitors amprenavir, atazanavir, indinavir, lopinavir, nelfnavir, ritonavir and saquinavir plus M8 nelfinavir metabolite and the non-nucleoside reverse transcriptase inhibitors efavirenz and nevirapine in human plasma by reversed–phase liquid chromatography. Ther Drug Monit 2005;2:186-92.

Choi So, Rezk NL, Kashuba AD. High-performance liquid chromatography assay fro the determination of the HIV–protease inhibitor tipranavir in human plasma in combination with nine other antiretroviral medications. J Pharm Biomed Anal 2007;43:1562-7.

Albert V, Modamio P, Lastra CF, Marino EL. Determination of saquinavir and ritonavir in human plasma by reversed–phase high–performance liquid chromatography and the analytical error function. J Pharm Biomed Anal 2004;36:835-40.

D’Avolio A, Baietto L, Siccardi M, Sciandra M, Simiele M. An HPLC-PDA method for the simultaneous quantification of the HIV integrase inhibitors raltegravir, the new non–nucleoside reverse transcriptase inhibitors etravirine, and 11 other antiretroviral agents in the plasma of HIV–infected patients. Ther Drug Monit 2008;30:662-9.

Rouzes A, Berthoin K, Xuereb F, Djabarouti S, Pellegrin I, Coupet AC, et al. Simultaneous determination of the antiretroviral agents: amprenavir, lopinavir, ritonavir, saquinavir and efavirenz in human peripheral blood mononuclear cells by high–performance liquid chromatography–mass spectrometry. J Chromatogr B: Analyt Technol Biomed Life Sci 2004;813:209-16.

Notari S, Bocedi A, Ippolito G, Narciso P, Pucillo LP, Tossini G, et al. Simultaneous determination of 16 anti-HIV drugs in human plasma by high-performance liquid chromatography. J Chromatogr B: Analyt Technol Biomed Life Sci 2006;831:258-66.

Dailly E, Raffi F, Jolliet P. Determination of atazanavir and other antiretroviral drugs (indinavir, amprenavir, nelfianvir and its active metabolite M8, saquinavir, ritonavir, lopinavir, nevirapine and efavirenz) plasma levels by high performance liquid chromatography with UV diction. J Chromatogr B: Analyt Technol Biomed Life Sci 2004;813:353-8.

Rezk NL, Tidwell RP, Kashuba AD. High–performance liquid chromatography assay for the quantification of HIV protease inhibitors and non–nucleoside reverse transcriptase inhibitors in human plasma. J Chromatogr B Analyt Technol Biomed Life Sci 2004;805:241-7.

Rebiere H, MKazel B, Civade C, Bonnet PA. Determination of 19 antiretroviral agents in pharmaceuticals or suspected products with two methods using high-performance liquid chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 2007;850:376-83.

Ofotokun, Ighovwerha, Edward P Acosta, Jeffrey L Lennox. Pharmacokinetics of an indinavir-ritonavir-fosamprenavir regimen in patients with human immunodeficiency virus. Pharmacother: J Human Pharmacol Drug Ther 2008;1:74-81.

la Porte CJL, Colbers EPH, Bertz R, Voncken DS, Wikstrom K. Pharmacokinetics of adjusted-dose lopinavir-ritonavir combined with rifampin in healthy volunteers. Antimicrob Agents Chemother 2004;48:1553–60.

Kityo C, Walker AS, Dickinson L, Lutwama F, Kayiwa J, Ssali F, et al. Pharmacokinetics of lopinavir-ritonavir with and without nonnucleoside reverse transcriptase inhibitors in ugandan HIV-Infected adults. Antimicrob Agents Chemother 2010;54:2965–73.

Marta Boffito, Akil Jackson, Alieu Amara, David Back, Saye Khoo, Chris Higgs, et al. Pharmacokinetics of once-daily darunavir-ritonavir and atazanavir-ritonavir over 72 H following drug cessation. Antimicrob Agents Chemother 2011;5:218–23.

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: Analyt Technol Biomed Life Sci 2005;816:233–42.

Chi J, Jayewardene AL, Stone JA, Motoya T, Aweeka FT. Simultaneous determination of five HIV protease inhibitors nelfinavir, indinavir, ritonavir, saquinavir and amprenavir in human plasma by LC/MS/MS. J Pharm Biomed Anal 2002;30:675–84.

Jiangeng Huanga, Nagsen Gautama, Sai Praneeth R Bathenaa, Upal Royb, JoEllyn McMillanb, Howard E Gendelmanb, et al. UPLC-MS/MS quantification of nanoformulated ritonavir, indinavir, atazanavir, and efavirenz in mouse serum and tissues. J Chromatogr B: Analyt Technol Biomed Life Sci 2011;879:2332–8.

Else L, Watson V, Tjia J, Hughes A, Siccardi M, Khoo S, et al. Validation of a rapid and sensitive high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) assay for the simultaneous determination of existing and new antiretroviral compounds. J Chromatogr B Anal Technol Biomed Life Sci 2010;878:1455–65.

Jeroen JA, van Kampen, Mariska L Reedijk, Peter C Burgers. Ultra-fast analysis of plasma and intracellular levels of HIV protease inhibitors in children: a clinical application of MALDI mass spectrometry. PLoS One 2010;5:11409.

Roland JW Meesters, Jeroen JA van Kampen, Mariska L Reedijk. Ultrafast and high-throughput mass spectrometric assay for therapeutic drug monitoring of antiretroviral drugs in pediatric HIV-1 infection applying dried blood spots. Anal Bioanal Chem 2010;398:319–28.

ICH Harmonised Tripartite Guideline. Validation of analytical procedures: Text and Methodology Q2. Courtesy:

Somatek Inc, USA; 2005.

Manosuthi W, Kiertiburanakul S, Amornnimit W, Prasithsirikul W, Thongyen S, Nilkamhang S, et al. Treatment outcomes and plasma level of ritonavir-boosted lopinavir monother-apy among HIV-infected patients who had NRTI and NNRTI failure. AIDS Res Ther 2009;6:30.

Josephson F, Andersson MC, Flamholc L, Gisslén M, Hagberg L, Ormaasen V, et al. The relation be-tween treatment outcome and efavir enz, atazanavir or lopinavir ex-posure in the NORTHIV trial of treatment-naive HIV-1 infected pa-tients. Eur J Clin Pharmacol 2010;66:349–57.

Hsu A, Isaacson J, Brun S, Bernstein B, Lam W, Bertz R, et al. Pharmacokinetic-pharmacodynamic analysis of lopinavir-ritonavir in combination with efavirenz and two nucleoside reverse transcriptase inhibitors in extensively pretreated human immunodeficiency virus-infected patients. Antimicrob Agents Chemother 2003;47:350–9.

Avolio A, Siccardi M, Sciandra M, Lorena B, Bonora S, Trentini L, et al. HPLC-MS method for the simultaneous quantification of the new HIV protease inhibitor darunavir, and 11 other antiretroviral agents in plasma of HIV-infected patients. J Chromatogr B: Analyt Technol Biomed Life Sci 2007;859:234–40.

Colombo S, Guignard N, Marzolini C, Telenti A, Biollaz J, Decosterd LA, et al. Determination of the new HIV-protease inhibitor atazanavir by liquid chromatography after solid-phase extraction. J Chromatogr B: Anal Technol Biomed Life Sci 2004;810:25–34.

Marzolini C, Beguin A, Telenti A, Schreyer A, Buclin T, Biollaz J, et al. Determination of lopinavir and nevirapine by high-performance liquid chromatography after solid-phase extraction: application for the assessment of their trans-placental passage at delivery. J Chromatogr B: Analyt Technol Biomed Life Sci 2002;774:127–40.

Published

01-07-2016

How to Cite

A. K., H. K., S. V., L. A., and G. Ramachandran. “A RAPID ISOCRATIC HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC-UV) METHOD FOR THE QUANTIFICATION OF RITONAVIR IN HUMAN PLASMA”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 8, no. 7, July 2016, pp. 64-68, https://innovareacademics.in/journals/index.php/ijpps/article/view/10276.

Issue

Section

Original Article(s)