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Anal Bioanal Chem. 2018 Nov;410(29):7773-7781. doi: 10.1007/s00216-018-1394-y. Epub 2018 Oct 2.

Development and validation of an LC-MS/MS assay for the quantification of dolutegravir extracted from human hair.

Author information

1
Division of Pharmacotherapy and Experimental Therapeutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, 1099C Genetic Medicine Building, CB# 7361, 120 Mason Farm Road, Chapel Hill, NC, 27599, USA. craig_sykes@unc.edu.
2
Division of Pharmacotherapy and Experimental Therapeutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, 1099C Genetic Medicine Building, CB# 7361, 120 Mason Farm Road, Chapel Hill, NC, 27599, USA.
3
Department of Clinical Pharmacy, University of California, San Francisco, School of Pharmacy, San Francisco, California, 94117, USA.

Abstract

Measurement of drug concentrations in hair provides a non-invasive approach to assess drug adherence. Here, we report on the development and validation of a method for the quantification of the antiretroviral dolutegravir (DTG) extracted from human hair. DTG is extracted from hair samples by sonication and incubation in 50:50 methanol:acetonitrile with 2% formic acid overnight at 40 °C. Following extraction, samples are analyzed by reverse-phase chromatography on a Waters Atlantis T3 (50 × 2.1 mm, 3-μm particle size) column with subsequent detection by electrospray ionization in positive ion mode on an AB Sciex API-5000 triple quadrupole mass spectrometer. The stable, isotopically labeled 13C,d5-DTG is used as an internal standard in the assay. The calibration range is 5-10,000 pg DTG/mL of extraction solvent with the ability to extract between 1 and 10 mg of hair/mL of extraction solvent. The assay was linear, accurate (inter-assay %bias within ± 6.5%), and precise (inter-assay %CV ≤ 10.3%). The assay was successfully used to analyze clinical samples from subjects on DTG regimens. Analysis of clinical samples suggested the potential presence of a degradation product, which was subsequently confirmed to occur with exposure to sunlight. The degradation of DTG could complicate absolute interpretation of clinical results, but the presence of this degradation product is easily evaluated with this assay to aid in data interpretation.

KEYWORDS:

Adherence; Antiretroviral; Dolutegravir; Hair; Validation

PMID:
30280227
PMCID:
PMC6501839
[Available on 2019-11-01]
DOI:
10.1007/s00216-018-1394-y
[Indexed for MEDLINE]

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