Validation and clinical application of a method to quantify efavirenz in cervicovaginal secretions from flocked swabs using liquid chromatography tandem mass spectrometry.


Olagunju, Adeniyi ORCID: 0000-0002-6588-5749, Nwogu, Jacinta ORCID: 0000-0002-0640-7165, Eniayewu, Oluwasegun, Atoyebi, Shakir ORCID: 0000-0002-1393-3753, Amara, Alieu ORCID: 0000-0002-1137-2948, Kpamor, John, Bolaji, Oluseye ORCID: 0000-0002-8295-7150, Adejuyigbe, Ebunoluwa, Owen, Andrew ORCID: 0000-0002-9819-7651 and Khoo, Saye ORCID: 0000-0002-2769-0967 (2021) Validation and clinical application of a method to quantify efavirenz in cervicovaginal secretions from flocked swabs using liquid chromatography tandem mass spectrometry. Wellcome open research, 6. 246-.

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Abstract

<b>Background</b> <b>:</b> A liquid chromatography tandem mass spectrometry method to quantify drugs in dried cervicovaginal secretions from flocked swabs was developed and validated using the antiretroviral efavirenz as an example. <b>Methods:</b> Cervicovaginal swabs (CVS) were prepared by submerging flocked swabs in efavirenz-spiked plasma matrix. Time to full saturation, weight uniformity, recovery and room temperature stability were evaluated. Chromatographic separation was on a reverse-phase C18 column by gradient elution using 1mM ammonium acetate in water/acetonitrile at 400 µL/min. Detection and quantification were on a TSQ Quantum Access triple quadrupole mass spectrometer operated in negative ionisation mode. The method was used to quantify efavirenz in CVS samples from human immunodeficiency virus (HIV)-positive women in the VADICT study (NCT03284645). A total of 98 samples (35 paired intensive CVS and DBS pharmacokinetic samples, 14 paired sparse CVS and DBS samples) from 19 participants were available for this analysis. <b>Results:</b> Swabs were fully saturated within 15 seconds, absorbing 128 µL of plasma matrix with coefficient of variation (%CV) below 1.3%. The method was linear with a weighting factor (1/X) in the range of 25-10000 ng/mL with inter- and intra-day precision (% CV) of 7.69-14.9%, and accuracy (% bias) of 99.1-105.3%. Mean recovery of efavirenz from CVS was 83.8% (%CV, 11.2) with no significant matrix effect. Efavirenz remained stable in swabs for at least 35 days after drying and storage at room temperature. Median (range) CVS efavirenz AUC <sub>0-24h</sub> was 16370 ng*h/mL (5803-22088), C <sub>max</sub> was 1618 ng/mL (610-2438) at a T <sub>max</sub> of 8.0 h (8.0-12), and C <sub>min</sub> was 399 ng/mL (110-981). Efavirenz CVS:plasma AUC <sub>0-24h</sub> ratio was 0.41 (0.20-0.59). <b>Conclusions:</b> Further application of this method will improve our understanding of the pharmacology of other therapeutics in the female genital tract, including in low- and middle-income countries.

Item Type: Article
Uncontrolled Keywords: Cervicovaginal fluid, Efavirenz, LC-MS/MS, Pharmacokinetics, Swab
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User: Symplectic Admin
Date Deposited: 07 Jun 2022 09:51
Last Modified: 16 Feb 2023 22:26
DOI: 10.12688/wellcomeopenres.17202.2
Open Access URL: https://wellcomeopenresearch.org/articles/6-246/v2
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3155992
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