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

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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.

Therapeutic drug monitoring using hair

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References

  1. Osterberg L, Blaschke T (2005) Adherence to medication. New Engl J Med 353:487–497

    Article  CAS  Google Scholar 

  2. Cramer J, Rosenheck R, Kirk G, Krol W, Krystal J (2003) Medication compliance feedback and monitoring in a clinical trial: predictors and outcomes. Value Health 6:566–573

    Article  Google Scholar 

  3. Ickovics J, Meisler AW (1997) Adherence in AIDS clinical trials: a framework for clinical research and clinical care. J Clin Epidemiology 50:385–391

    Article  CAS  Google Scholar 

  4. Vanhove GF, Schapiro JM, Winters MA, Merigan TC, Blaschke TF (1996) Patient compliance and drug failure in protease inhibitor monotherapy. JAMA 276:1955–1956

    Article  CAS  Google Scholar 

  5. Paterson DL, Swindells S, Mohr J, Brester M, Vergis EN, Squier C, Wagener MM, Singh N (2000) Adherence to protease inhibitor therapy and outcomes in patients with HIV infection. Ann Intern Med 133:21–30

    CAS  Google Scholar 

  6. Condra JH, Schleif WA, Blahy OM, Gabryelski LJ, Graham DJ, Quintero JC, Rhodes A, Robbins HL, Roth E, Shivaprakash M, Titus D, Yang T, Tepplert H, Squires KE, Deutsch PJ, Emini EA (1995) In vivo emergence of HIV-1 variants resistant to multiple protease inhibitors. Nature 374:569–571

    Article  CAS  Google Scholar 

  7. Salomon JA, Hogan DR, Stover J, Stanecki KA, Walker N, Ghys PD, Schwartländer B (2005) Integrating HIV prevention and treatment: from slogans to impact. PLoS Med 2:e16

    Article  Google Scholar 

  8. Castilla J, Del Romero J, Hernando V, Marincovich B, García S, Rodríguez C (2005) Effectiveness of highly active antiretroviral therapy in reducing heterosexual transmission of HIV. J AIDS 40:96–101

    Google Scholar 

  9. Podsadecki TJ, Vrijens BC, Tousset EP, Rode RA, Hanna GJ (2008) “White coat compliance” limits the reliability of therapeutic drug monitoring in HIV-1-infected patients. HIV Clin Trials 9:238–246

    Article  Google Scholar 

  10. Khoo SH, Lloyd J, Dalton M, Bonington A, Hart E, Gibbons S, Flegg P, Sweeney J, Wilkins EG, Back DJ (2006) Pharmacologic optimization of protease inhibitors and nonnucleoside reverse transcriptase inhibitors (POPIN)—a randomized controlled trial of therapeutic drug monitoring and adherence support. J AIDS 41:461–467

    CAS  Google Scholar 

  11. Burger DM, Hugen PW, Aarnoutse RE, Hoetelmans RM, Jambroes M, Nieuwkerk PT, Schreij G, Schneider MM, van der Ende ME, Lange JM (2003) Treatment failure of nelfinavir-containing triple therapy can largely be explained by low nelfinavir plasma concentrations. Ther Drug Monit 25:73–80

    Article  CAS  Google Scholar 

  12. Clevenbergh P, Garraffo R, Durant J, Dellamonica P (2002) PharmAdapt: a randomized prospective study to evaluate the benefit of therapeutic monitoring of protease inhibitors: 12 week results. AIDS 16:2311–2315

    Article  CAS  Google Scholar 

  13. Back D, Gatti G, Fletcher C, Garaffo R, Haubrich R, Hoetelmans R, Kurowski M, Luber A, Merry C, Perno CF (2002) Therapeutic drug monitoring in HIV infection: current status and future directions. AIDS 16(Suppl 1):S5–S37

    Article  CAS  Google Scholar 

  14. Back D, Gibbons S, Khoo S (2006) An update on therapeutic drug monitoring for antiretroviral drugs. Ther Drug Monit 28:468–473

    Article  Google Scholar 

  15. Nettles RE, Kieffer TL, Parsons T, Johnson J, Cofrancesco J Jr, Gallant JE, Carson KA, Siliciano RF, Flexner C (2006) Marked intraindividual variability in antiretroviral concentrations may limit the utility of therapeutic drug monitoring. Clin Infec Dis 42:1189–1196

    Article  CAS  Google Scholar 

  16. Nakahara Y (1999) Hair analysis for abused and therapeutic drugs. J Chromatogr B Biomed Sci Appl 733:161–180

    Article  CAS  Google Scholar 

  17. Uematsu T (1993) Therapeutic drug monitoring in hair samples. Principles and practice. Clin Pharmacokin 25:83–87

    Article  CAS  Google Scholar 

  18. Gandhi M, Greenblatt RM (2002) Hair it is: the long and short of monitoring antiretroviral treatment. Ann Intern Med 137:696–697

    Google Scholar 

  19. Beumer JH, Bosman IJ, Maes RA (2001) Hair as a biological specimen for therapeutic drug monitoring. Int J Clin Pract 55:353–357

    CAS  Google Scholar 

  20. Bernard L, Vuagnat A, Peytavin G, Hallouin MC, Bouhour D, Nguyen TH, Vildé JL, Bricaire F, Raguin G, de Truchis P, Ghez D, Duong M, Perronne C (2002) Relationship between levels of indinavir in hair and virologic response to highly active antiretroviral therapy. Ann Intern Med 137:656–659

    CAS  Google Scholar 

  21. Servais J, Peytavin G, Arendt V, Staub T, Schneider F, Hemmer R, Burtonboy G, Schmit JC (2001) Indinavir hair concentration in highly active antiretroviral therapy-treated patients: association with viral load and drug resistance. AIDS 15:941–943

    Article  CAS  Google Scholar 

  22. Duval X, Peytavin G, Breton G, Ecobichon JL, Descamps D, Thabut G, Leport C (2007) Hair versus plasma concentrations as indicator of indinavir exposure in HIV-1-infected patients treated with indinavir/ritonavir combination. AIDS 21:106–108

    Article  CAS  Google Scholar 

  23. Huang Y, Gandhi M, Greenblatt RM, Gee W, Lin ET, Messenkoff N (2008) Sensitive analysis of anti-HIV drugs, efavirenz, lopinavir and ritonavir, in human hair by liquid chromatography coupled with tandem mass spectrometry. Rapid Commun Mass Spectrom 22:3401–3409

    Article  CAS  Google Scholar 

  24. Gandhi M, Ameli N, Bacchetti P, Gange SJ, Anastos K, Levine A, Hyman CL, Cohen M, Young M, Huang Y, Greenblatt RM (2009) Protease inhibitor levels in hair strongly predict virologic response to treatment. AIDS 23:471–478

    Article  CAS  Google Scholar 

  25. Gandhi M, Ameli N, Bacchetti P, Anastos K, Gange SJ, Minkoff H, Young M, Milam J, Cohen MH, Sharp GB, Huang Y, Greenblatt RM (2011) Atazanavir concentration in hair is the strongest predictor of outcomes on antiretroviral therapy. Clin Infect Dis 52:1267–1275

    Google Scholar 

  26. Gandhi M, Ameli N, Bacchetti P, Huang Y, Gange SJ, Anastos K, Levine A, Cohen M, Young M, Greenblatt RM (2009) Concentrations of Efavirenz in Hair Are Strongly Correlated with Virologic Response. 16th Conference on Retroviruses and Opportunistic Infections (CROI), Montreal, Canada, February 8–11

  27. van Zyl GU, van Mens TE, Mcilleron H, Zeier M, Nachega JB, Decloedt E, Malavazzi C, Smith P, Huang Y, van der Merwe L, Gandhi M, Maartens G (2011) Low lopinavir plasma or hair concentrations explain second-line protease inhibitor failures in a resource-limited setting. J AIDS 56:333–339

    Google Scholar 

  28. Murphy R, Montaner J (1996) Nevirapine: a review of its development, pharmacological profile and potential for clinical use. J Expert Opinion on Investigational Drugs 5:1183–1199

    Article  CAS  Google Scholar 

  29. Sullivan JL (2003) Prevention of mother-to-child transmission of HIV–what next? J AIDS 34(Suppl 1):S67–S72

    Google Scholar 

  30. Duong M, Buisson M, Peytavin G, Kohli E, Piroth L, Martha B, Grappin M, Chavanet P, Portier H (2005) Low trough plasma concentrations of nevirapine associated with virologic rebounds in HIV-infected patients who switched from protease inhibitors. Ann Pharmacother 39:603–609

    Article  CAS  Google Scholar 

  31. Gonzalez de Requena D, Nunez M, Jimenez-Nacher I, Soriano V (2002) Liver toxicity caused by nevirapine. AIDS 16:290–291

    Article  Google Scholar 

  32. McKoy JM, Bennett CL, Scheetz MH, Differding V, Chandler KL, Scarsi KK, Yarnold PR, Sutton S, Palella F, Johnson S, Obadina E, Raisch DW, Parada JP (2009) Hepatotoxicity associated with long- versus short-course HIV-prophylactic nevirapine use: a systematic review and meta-analysis from the Research on Adverse Drug events And Reports (RADAR) project. Drug Saf 32:147–158

    Article  CAS  Google Scholar 

  33. Martínez E, Blanco JL, Arnaiz JA, Pérez-Cuevas JB, Mocroft A, Cruceta A, Marcos MA, Milinkovic A, García-Viejo MA, Mallolas J, Carné X, Phillips A, Gatell JM (2001) Hepatotoxicity in HIV-1-infected patients receiving nevirapine-containing antiretroviral therapy. AIDS 15:1261–1268

    Article  Google Scholar 

  34. Barkan SE, Melnick SL, Preston-Martin S, Weber K, Kalish LA, Miotti P, Young M, Greenblatt R, Sacks H, Feldman J (1998) The Women’s Interagency HIV Study. WIHS Collaborative Study Group. Epidemiology 9:117–125

    Article  CAS  Google Scholar 

  35. Viswanathan CT, Bansal S, Booth B, DeStefano AJ, Rose MJ, Sailstad J, Shah VP, Skelly JP, Swann PG, Weiner R (2007) Workshop/Conference Report-Quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. AAPS J 9:E30–E42

    Article  Google Scholar 

  36. Spooner N (2010) A glowing future for dried blood spot sampling. Bioanalysis 2:1343–1344

    Article  CAS  Google Scholar 

  37. Amsterdam P, Waldrop C (2010) The application of dried blood spot sampling in global clinical trials. Bioanalysis 2:1783–1786

    Article  Google Scholar 

Download references

Acknowledgment

We thank Richard Bonderud for his critical review of this manuscript. We thank the WIHS participants who contributed hair specimens for this study. This study is supported by NIH/NIAID Grant R01 AI 065233- and also U01AI034989.

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Authors and Affiliations

  1. Drug Studies Unit, Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, 94143, USA

    Yong Huang, Qiyun Yang, Kwangchae Yoon, Yvonne Lei, Robert Shi, Winnie Gee & Emil T. Lin

  2. Department of Medicine, University of California, San Francisco, CA, 94143, USA

    Ruth M. Greenblatt & Monica Gandhi

  3. Department of Clinical Pharmacy, University of California, San Francisco, CA, 94143, USA

    Ruth M. Greenblatt

  4. Department of Epidemiology and Biostatistics, University of California, San Francisco, CA, 94143, USA

    Ruth M. Greenblatt

  5. Department of Bioengineering and Therapeutic Sciences, School of Pharmacy, University of California San Francisco, San Francisco, CA, 94143-0446, USA

    Yong Huang

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  1. Yong Huang
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Correspondence to Yong Huang.

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Huang, Y., Yang, Q., Yoon, K. et al. Microanalysis of the antiretroviral nevirapine in human hair from HIV-infected patients by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 401, 1923–1933 (2011). https://doi.org/10.1007/s00216-011-5278-7

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  • DOI: https://doi.org/10.1007/s00216-011-5278-7

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