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

  • Yong Huang
  • Qiyun Yang
  • Kwangchae Yoon
  • Yvonne Lei
  • Robert Shi
  • Winnie Gee
  • Emil T. Lin
  • Ruth M. Greenblatt
  • Monica Gandhi
Original Paper


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.


Therapeutic drug monitoring using hair


Antiretroviral drug Nevirapine Hair LC-MS/MS TDM Adherence 

Supplementary material

216_2011_5278_MOESM1_ESM.pdf (194 kb)
ESM 1(PDF 194 kb)


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Yong Huang
    • 1
    • 5
  • Qiyun Yang
    • 1
  • Kwangchae Yoon
    • 1
  • Yvonne Lei
    • 1
  • Robert Shi
    • 1
  • Winnie Gee
    • 1
  • Emil T. Lin
    • 1
  • Ruth M. Greenblatt
    • 2
    • 3
    • 4
  • Monica Gandhi
    • 2
  1. 1.Drug Studies Unit, Department of Bioengineering and Therapeutic SciencesUniversity of CaliforniaSan FranciscoUSA
  2. 2.Department of MedicineUniversity of CaliforniaSan FranciscoUSA
  3. 3.Department of Clinical PharmacyUniversity of CaliforniaSan FranciscoUSA
  4. 4.Department of Epidemiology and BiostatisticsUniversity of CaliforniaSan FranciscoUSA
  5. 5.Department of Bioengineering and Therapeutic Sciences, School of PharmacyUniversity of California San FranciscoSan FranciscoUSA

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