Original Paper

Analytical and Bioanalytical Chemistry

, 401:1923

First online:

Microanalysis of the antiretroviral nevirapine in human hair from HIV-infected patients by liquid chromatography-tandem mass spectrometry

  • Yong HuangAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of CaliforniaDepartment of Bioengineering and Therapeutic Sciences, School of Pharmacy, University of California San Francisco Email author 
  • , Qiyun YangAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California
  • , Kwangchae YoonAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California
  • , Yvonne LeiAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California
  • , Robert ShiAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California
  • , Winnie GeeAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California
  • , Emil T. LinAffiliated withDrug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California
  • , Ruth M. GreenblattAffiliated withDepartment of Medicine, University of CaliforniaDepartment of Clinical Pharmacy, University of CaliforniaDepartment of Epidemiology and Biostatistics, University of California
  • , Monica GandhiAffiliated withDepartment of Medicine, University of California

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Abstract

Sufficient drug exposure is crucial for maintaining durable responses to HIV treatments. However, monitoring drug exposure using single blood samples only provides short-term information and is highly subject to intra-individual pharmacokinetic variation. Drugs can accumulate in hair over a long period of time, so hair drug levels can provide drug exposure information over prolonged periods. We now report on a specific, sensitive, and reproducible liquid chromatography-tandem mass spectrometry method for measuring nevirapine (NVP), a widely used antiretroviral drug, levels in human hair using even a single short strand of hair. Hair samples are cut into small segments, and the drug is extracted in methanol/trifluoroacetic acid (v/v, 9:1) shaken at 37 °C in a water bath overnight, followed by liquid–liquid extraction under alkaline conditions. The extracted samples are then separated on a BDS-C18 column with a mobile phase composed of 50% acetonitrile containing 0.15% acetic acid and 4 mM ammonium acetate with an isocratic elution for a total run time of 3 min and detected by triple quadrupole electrospray multiple reaction mode at precursor/product ion at 267.0 > 225.9 m/z. Deuterated nevirapine-d5 was used as an internal standard. This method was validated from 0.25 to 100 ng/mg using 2 mg hair samples. The accuracies for spiked NVP hair control samples were 98–106% with coefficients of variation (CV) less than 10%. The CV for incurred hair control samples was less than 7%. The extraction efficiency for incurred control hair samples was estimated at more than 95% by repeated extractions. This method has been successfully applied to analyze more than 1,000 hair samples from participants in a large ongoing cohort study of HIV-infected participants. We also showed that NVP in human hair can easily be detected in a single short strand of hair. This method will allow us to identify drug non-adherence using even a single strand of hair.

https://static-content.springer.com/image/art%3A10.1007%2Fs00216-011-5278-7/MediaObjects/216_2011_5278_Figa_HTML.gif
Figure

Therapeutic drug monitoring using hair

Keywords

Antiretroviral drug Nevirapine Hair LC-MS/MS TDM Adherence