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

Abstract

The adherence assessment based on the combination of nevirapine (NVP) and its two metabolites (2-hydroxynevirapine and 3-hydroxynevirapine) would more comprehensively and accurately reflect long-term adherence than that of a single prototype. This study aimed to develop a specific, sensitive and selective method for simultaneous detection of the three compounds in hair and explore whether there was consistency among the three compounds in assessing long-term adherence. Furthermore, 75 HIV-positive patients who were taking the NVP drug were randomly recruited and divided into two groups (high-and low-adherence group). All participants self-reported their days of oral drug administration per month and provided their hair strands closest to the scalp at the region of posterior vertex. The concentrations of three compounds in the hair were determined using a developed LC-MS/MS method in multiple reaction monitoring. This method showed good performances in limit of quantification and accuracy with the recoveries from 85 to 115% and in precision with the intra-day and inter-day coefficients of variation within 15% for the three compounds. The population analysis revealed that patients with high-adherence showed significantly higher concentrations than those with low-adherence for all three compounds. There were significantly moderate correlations of nevirapine with 2-hydroxynevirapine and 3-hydroxynevirapin and high correlation between 2-hydroxynevirapine and 3-hydroxynevirapin. The two NVP’s metabolites showed high consistency with NVP in evaluating long-term adherence.

Association of Hair Concentrations of Antiretrovirals with Virologic Outcomes Among People Living with HIV in Guangxi, China

“…Then, hair sample was chopped to 1–2 mm length segments with scissors, and 10 mg weighed, processed, and analyzed using high-performance liquid chromatography (Agilent 1200 HPLC system, Agilent, Waldbronn, Germany) and tandem mass spectrometry (ABI 3200Qtrap, ABI, Foster City, CA, USA) (LC-MS/MS) as described in previous studies. 21 , 24 , 40 Briefly, following the 2013 FDA guidelines, antiretrovirals in the cut hair samples were extracted with methanol and internal standard in a 37°C shaking water bath overnight (>16 hours) and then analyzed by an LC-MS/MS. Standard curves were linear in the range of 6–40,000 pg for 3TC, 10–40,000 pg for AZT, and 12–40,000 pg for EFV with good linearity and reproducibility.…”

Clinical Value of Emerging Bioanalytical Methods for Drug Measurements: A Scoping Review of Their Applicability for Medication Adherence and Therapeutic Drug Monitoring

“…Therefore, white-coat adherence, which is the intake of medication just before a clinical visit, cannot be detected. For hair, the absence of the drug would mean the drug was not or was only very limitedly taken for longer periods [ 33 , 57 , 64 , 65 , 78 ].…”

“…No reported assays were infeasible for MAM. Furthermore, highly valid methods were available for 16 drugs measured for MAM mostly in epilepsy, HIV/AIDS, and pain [21,52,[54][55][56][57][58][59][60][61][62][63][64][65]. Hair is the most widely used method to detect the presence of 37 unique drugs and/or their metabolites, followed by saliva (31 drugs) and DBS (20 drugs).…”

Development of an improved method to estimate the days of continuous drug ingestion, based on the micro‐segmental hair analysis

“…The improved method for estimating the days of continuous drug ingestion would be applicable to not only investigate drug-related crimes but also check the adherence to drug regime. [30][31][32]…”

“…In future studies, more data should be acquired using hair strands collected from drug abusers and patients who were administered various prescribed medicines. The improved method for estimating the days of continuous drug ingestion would be applicable to not only investigate drug‐related crimes but also check the adherence to drug regime 30–32 …”