Simultaneous determination of antiretroviral drugs in human hair with liquid chromatography-electrospray ionization-tandem mass spectrometry

Abstract

The determination of the concentrations of antiretroviral drugs in hair is believed to be an important means for the assessment of the long-term adherence to highly active antiretroviral therapy. At present, the combination of tenofovir, lamivudine and nevirapine is widely used in China. However, there was no research reporting simultaneous determination of the three drugs in hair. The present study aimed to develop a sensitive method for simultaneous determination of the three drugs in 2-mg and 10-mg natural hair (Method 1 and Method 2). Hair samples were incubated in methanol at 37 °C for 16 h after being rinsed with methanol twice. The analysis was performed on high performance liquid chromatography tandem mass spectrometry with electronic spray ionization in positive mode and multiple reactions monitoring. Method 1 and Method 2 showed the limits of detection at 160 and 30 pg/mg for tenofovir, at 5 and 6 pg/mg for lamivudine and at 15 and 3 pg/mg for nevirapine. The two methods showed good linearity with the square of correlation coefficient >0.99 at the ranges of 416-5000 and 77-5000 pg/mg for tenofovir, 12-5000 and 15-5000 pg/mg for lamivudine and 39-50,000 and 6-50,000 pg/mg for nevirapine. They gave intra-day and inter-day coefficient of variation <15% and the recoveries ranging from 80.6 to 122.3% and from 83.1 to 114.4%. Method 2 showed LOD and LOQ better than Method 1 for tenofovir and nevirapine and matched Method 1 for lamivudine, but there was high consistency between them in the determination of the three drugs in hair. The population analysis with Method 2 revealed that the concentrations in hair were decreased with the distance of hair segment away from the scalp for the three antiretroviral drugs.

Comparing self-reported medication adherence measures with hair antiretroviral concentration among people living with HIV in Guangxi, China

“…The proximal section of the hair sample (about 1 cm, reflecting the last month of TDF dosing) was rinsed with methanol and dried under a blow of pure nitrogen gas. Then hair sample was chopped to 1-2 mm length segments with scissors, and 10 mg weighed, processed, and analyzed using highperformance liquid chromatography (Agilent 1200 HPLC system, Agilent, Waldbronn, Germany) and tandem mass spectrometry (ABI 3200Qtrap, ABI, Foster City, CA, USA) (LC-MS/MS) [40,41]. Briefly, following the 2013 FDA guideline, The tenofovir in the cut hair sample was extracted with methanol and internal standard in a 37 °C shaking water bath overnight (> 16 h) and then analyzed by an LC-MS/MS [40].…”

LC-MS/MS Quantification of Nevirapine and Its Metabolites in Hair for Assessing Long-Term Adherence

“…Additionally, there is large inter-individual difference in drug metabolism. For example, coefficients of variation (CV) in the metabolism of NVP, lamivudine and tenofovir were 99.1, 75.4 and 103.8% as shown by the previous study [30]. Therefore, the adherence assessment based on the drug prototype in hair might be less accurate.…”

Application of High-Performance Liquid Chromatography for Simultaneous Determination of Tenofovir and Creatinine in Human Urine and Plasma Samples

“…Testing of TFV level in human peripheral blood might be painful for patients and carries the risk of additional HIV/HBV infections. Methods created for TFV determination in non-infected matrices, such as urine [28,30] or hair [31,32], do not provide the possibility for simultaneous monitoring concentration of Crn. Previously published reports mainly described methods for the determination of TFV in plasma [25,26,29,[33][34][35][36][37][38][39][40], which determines a higher risk of infection.…”

“…Previously described assays dedicated for TFV determination are based on RP-HPLC, usually coupled with mass spectrometry [25,29,32,37,39,40]. Methods requiring commonly available detectors such as spectrophotometric [35,36] or spectrofluorimetric [33,34] detectors are used rarely.…”