Development and validation of a high performance liquid chromatography method to determine nevirapine in plasma in a resource-limited setting

Abstract

Background There are several instances where nevirapine pharmacokinetic monitoring may be useful, such as in special populations or pharmacokinetic drug interaction studies that require the ascertainment of nevirapine pharmacokinetics in the sub-Saharan region. Objectives The main aim of this study was to produce a validated, sustainable and relevant nevirapine assay method that meets bio-analytical regulatory requirements. Methods The developed method utilised a Waters 2795 Alliance high performance liquid chromatography system with a 2996 photo diode array detector, an Atlantis dC18 5 micron, 3.9 mm × 150 mm analytical column and a gradient flow rate of 1 mL/min. Ultraviolet detection data were collected from 210 nm to 400 nm, extracted at 260 nm, and processed for nevirapine and internal standard peak height responses. Results The method proved to be linear (R2 0.995), precise (+1.92% – +9.69%) and accurate (-9.70% – 12.0%). Recovery for the analyte and internal standard was between 98.8% and 114%. The method showed good specificity as no interferences were caused by common African traditional medicines, anti-tuberculosis medications or other concomitant antiretrovirals nor endogenous components. Conclusion The method is reproducible, relevant to our setting and uses considerably low plasma volumes with preservation of some consumables, a desirable key factor in a resource-limited setting.

Cross-validation of a high-performance liquid chromatography nevirapine plasma assay in a resource-limited setting in Zimbabwe

“…The UZ-IPSL measured nevirapine using a validated HPLC-ultraviolet assay detailed previously. 2 The UZ-IPSL chromatographic system consisted of a Shimadzu LC20A HPLC using ultraviolet photodiode array detection (model SPD-M20A) and LabSolutions Software (version 5.8; Kyoto, Japan). The assay quantitation range was 500 ng/mL to 15 000 ng/mL.…”

“…A5279 samples were assayed over five runs; the two last runs included samples that were above the upper limit of quantification and were diluted (1:8) according to the validated dilution protocol. 2 …”

“…The Clinical Pharmacology Quality Assurance programme, which monitors all PSLs within the AIDS Clinical Trials Group, provided technical guidance to and developed a comparative nevirapine validation study for the UZ-IPSL to analyse replicate HIV clinical samples. Validation of the nevirapine bioanalytics method 2 was essential at the time because of nevirapine use in the sub-Saharan region and results were generated by a reverse-phase high-performance liquid chromatography (HPLC) system using ultraviolet detection with gradient-elution separation. 3 , 4 , 5 This report compares nevirapine bioanalysis at two laboratories: the United States-based, University of Nebraska Medical Center Pharmacology Specialty Laboratory (UNMC-PSL) and the UZ-IPSL.…”

Two decades (1998 to 2018) of collaborative human immunodeficiency virus clinical pharmacology capacity building in a resource constrained setting

“…Chromatographic assays for antiretroviral drug quantitation were validated in the context of concomitant herbal medicine use. This contributed to capacity building for therapeutic drug monitoring of cART in a high herbal medicine use setting which will be valuable for both practice and research [ 18 ]. In addition, other publications highlighted potential and actual adverse drug events which may lead to poor adherence and the need for pharmacovigilance programs to expand to include herbal medicines [ 19 , 20 ].…”