Determination of Nevirapine Levels in Hair and Plasma Samples of People Living With HIV in Kenya Using HPLC-UV and LC-MS/MS Techniques
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Date
2025-01
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Kenyatta University
Abstract
Monitoring the response to antiretroviral drugs (ARVs) in patients is crucial for
effective HIV treatment. Antiretroviral therapy (ART) is pivotal in mitigating the
epidemic of human immunodeficiency virus (HIV), particularly in the prevention of
vertical transmission of the virus. Monitoring of the adherence of the patients to ART
is imperative for gauging their response to antiretroviral drugs (ARVs) and identifying
treatment inadequacies. Methods previously used in monitoring adherence, like
measuring ARV drug concentrations in blood and urine, are limited by their ability to
only reflect doses taken within 1 to 2 days before the day of sampling. In response to
these limitations, hair testing has emerged as a preferred tool for assessing chronic
exposure to various substances, owing to its extended detection window. Therefore,
this study determined the viability of hair samples for adherence monitoring, serving
as an alternative to blood-based ARV analysis. Furthermore, it investigated the
potential of using high-performance liquid chromatography with a UV detector
(HPLC-UV) as a cost-effective substitute of liquid chromatography-tandem mass
spectrometry (LC-MS/MS) technique, particularly in resource-limited settings like
Kenyan hospitals. The research focused on nevirapine (NVP), a fundamental
component of Kenya's first-line ART regimen. Hair and blood samples were collected
from consenting patients with varying viral loads. NVP levels in these samples were
compared using HPLC-UV and LC-MS/MS instruments. Significantly, the findings
reveal no substantial difference between hair and plasma NVP concentrations, as
verified by statistical tests. A robust positive association between the two
measurement methods further validates the utility of HPLC-UV for monitoring ARV
drug concentrations in both hair and blood in resource-limited settings. Quantitatively,
the median (IQR) NVP levels in hair and blood samples were 67.80 ng/mL and
706.50 ng/mL for HPLC-UV, and 36.80 ng/mL and 19.32 ng/mL for LC-MS/MS,
respectively. The Wilcoxon signed-rank test, yielding a statistical result of (Z = -0.93,
p > 0.05), confirms no significant difference between hair and plasma NVP
concentrations. The Spearman rank test indicates a significant positive association
between NVP concentrations analysed using both LC-MS/MS and HPLC-UV (r2
=
0.995, p < 0.05) for hair samples, and (r2
= 0.966, p < 0.05) for plasma samples. This
research demonstrated the potential of using hair testing as a non-invasive, cost effective means to quantitatively monitor ARV adherence among people living with
HIV, particularly in regions lacking traditional laboratory facilities and skilled
personnel for blood sampling. Importantly, it does not suggest replacing plasma
testing with hair testing in all scenarios; instead, these methods can complement each
other for a comprehensive assessment of adherence to antiretroviral therapy
Description
A Thesis Submitted in Partial Fulfillment of the Requirement for the Award of the Degree of Master of Science in Applied Analytical Chemistry the School of Pure and Applied Sciences of Kenyatta University January, 2025
Supervisors;
1.Margaret M. Nganga
2.Beatrice Irungu
2.Elizabeth Kigondu