High Performance Liquid Chromatographic Assay for the Determination of Protease Inhibitors (PIs) and Non-nucleoside Reverse Transcriptase Inhibitors (NNRTIs) in Human Plasma

Authors

  • Kantima Sangsiriwut Department of Preventive and Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University
  • Thanomsak Anekthananon Department of Preventive & Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University
  • Winai Ratanasuwan Department of Preventive & Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University
  • Surapol Suwanagool Department of Preventive & Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University
  • Teera Kolladarungkri Department of Preventive & Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University

Keywords:

non-nucleoside reverse transcriptase inhibitors, protease inhibitors, HPLC, HIV

Abstract

Objective: To develop and validate a high performance liquid chromatography (HPLC) method for simultaneous quantitative determination offive HIV protease inhibitors (PIs): indinavir (IDV), lopinavir (LPV), nelfinavir (NFV), ritonavir (RTV), saquinavir (SQV), andtwonon-nucleoside reverse transcriptase inhibitors (NNRTIs):nevirapine (NVP), andefavirenz (EFV) inhumanplasma.

Methods: A sample of 200 µL of plasma and an internal standard were extracted with tert-butyl methyl ether. The compounds were separated on a reversed-phase C18 column with gradient phase of 25 mM phosphate buffer (pH 4.9) and acetonitrile. The limit of quantation, accuracy, precision, specificity, stability andrecovery were tested.

Results: The lower limit of quantitation for all drugs was 75 ng/mL. The standard curve was linear in the range of 75 ng/mL to 20,000 ng/mL. Intra-day and inter-day variability ranged from 0.1% to 2.4% and 0.3% to 4.1%, respectively. Accuracy ranged from 98.4%-102.4% for three quality controls (75, 100, and 1,000 ng/mL) for all drugs measured. The extraction recovery ranged from 98.7%-101.3%.

Conclusion: This method provides a simple, accurate, and precise method for monitoring of plasma concentrations of five PIs and two NNRTIs in the case of weak economy and out of date instrumental limitations.

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Published

31-07-2020

How to Cite

Sangsiriwut, K., Anekthananon, T. ., Ratanasuwan, W. ., Suwanagool, S., & Kolladarungkri, T. (2020). High Performance Liquid Chromatographic Assay for the Determination of Protease Inhibitors (PIs) and Non-nucleoside Reverse Transcriptase Inhibitors (NNRTIs) in Human Plasma. Siriraj Medical Journal, 64(1), 1–6. Retrieved from https://he02.tci-thaijo.org/index.php/sirirajmedj/article/view/244137

Issue

Vol. 64 No. 1 (2012): Jan-Feb

Section

Original Article

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