Method Development for the Detection of Basic/Weak Basic Drugs in Hair by LCMSMS: Comparison between Methanolic and Alkaline Extraction on Real Samples


Detection of drugs in hair has become popular in recent years. The significantly long drug detection window (months) in hair has allowed the retrospective investigation and measurement of past consumption of drug. As the majority of drugs are basic, an extraction method was developed based on a methanolic solution for detection of basic/weak basic drugs in hair. It was compared with alkaline digestion (NaOH) followed by LLE. A filtration step with filtration vials was added and their materials were compared. After filtration, extracts were injected directly onto a C18 column coupled to Sciex ABI 2000 MSMS. The mobile phase was 50% methanol, 0.1% formic acid and 2 mM ammonium acetate (isocratic). Both methods were compared by applying them to real samples. Results showed that calibration was linear with r2 of 0.991-0.999 for 20 tested analytes. The matrix effect was assessed to be between 91.4%- 110.2% for 18 analytes. PTFE filter material showed better recoveries over the GMF and PVDF based filters. Stability of analytes during extraction in general was better with methanolic incubation rather than alkaline digestion. With regard to real sample recovery, 6 out of 10 analytes recovered better with alkaline digestion. In conclusion, the methanolic method is capable of extracting most basic drugs in hair samples but only part of the total incorporated drug. Therefore, these results suggest that a combination of both methods (methanolic and alkaline extractions) in hair sample processing for general detection of basic and weak basic drugs may produce better results. However, not all basic drugs are stable under alkaline digestion.

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J. Al Jaber, D. Holt and A. Johnston, "Method Development for the Detection of Basic/Weak Basic Drugs in Hair by LCMSMS: Comparison between Methanolic and Alkaline Extraction on Real Samples," Pharmacology & Pharmacy, Vol. 3 No. 3, 2012, pp. 263-274. doi: 10.4236/pp.2012.33035.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] I. M. Kempson and E. Lombi, “Hair Analysis as a Bio-monitor for Toxicology, Disease and Health Status,” Chemical Society Reviews, Vol. 40, No. 7, 2011, pp. 3915- 3940. doi:10.1039/c1cs15021a
[2] P. Kintz, “Analytical And Practical Aspects of Drug Testing in Hair,” CRC Press, New York, 2007.
[3] F. Pragst and M. A. Bali-kova, “State of the Art in Hair Analysis for Detection of Drug and Alcohol Abuse,” Clinica Chimica Acta, Vol. 370, No. 1-2, 2006, pp. 17-49. doi:10.1016/j.cca.2006.02.019
[4] R. Smolders, K. W. Schramm, M. Nickmilder and G. Schoeters, “Applicability of Non-Invasively Collected Matrices for Human Biomonitoring,” Environmental Health, Vol. 8, No. 1, 2009, p. 8. doi:10.1186/1476-069X-8-8
[5] F. Musshoff and B. Madea, “New Trends in Hair Analysis and Scientific Demands on Validation and Technical Notes,” Forensic Science International, Vol. 165, No. 2-3, 2007, pp. 204-215. doi:10.1016/j.forsciint.2006.05.024
[6] M. Villain, M. V. C. Concheiro and P. Kintz, “Screening Method For benzodiazepines and Hypnotics in Hair at pg/mg Level by Liquid Chromatography-Mass Spectro-metry/Mass Spectrometry,” Journal of Chromatography B, Vol. 825, No. 1, 2005, pp. 72-78. doi:10.1016/j.jchromb.2004.12.036
[7] M. Laloup, M. Ramirez Fernandez Mdel, G. De Boeck, M. Wood, V. Maes and N. Samyn, “Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for the Si-multaneous Determination of 26 Benzodiazepines and Metabolites, Zolpidem and Zopiclone, in Blood, Urine, and Hair,” Journal of Analytical Toxicology, Vol. 29, No. 7, 2005, pp. 616-26.
[8] F. Musshoff and B. Madea, “Analytical Pitfalls in Hair Testing,” Analytical and Bio-analytical Chemistry, Vol. 388, No. 7, 2007, pp. 1475-1494. doi:10.1007/s00216-007-1288-x
[9] F. J. Couper, I. M. McIntyre and O. H. Drummer, “Extraction of Psychotropic Drugs from Human Scalp Hair,” Journal of the Forensic Science, Vol. 40, No. 1, 1995, pp. 83-86.
[10] M. Josefsson, R. Kronstrand, J. Andersson and M. Roman, “Evaluation of Electrospray Ionisation Liquid Chromato-graphy-Tandem Mass Spectrometry for Rational Determination of a Number of Neuroleptics and Their Major Metabolites in Human Body Fluids and Tissues,” Journal of Chromatography B, Vol. 789, No. 1, 2003, pp. 151-167. doi:10.1016/S1570-0232(03)00207-1
[11] J. Villamor, A. Bermejo, P. Fernandez and M. Tabernero, “TECHNICAL NOTE: A New GC-MS Method for the Determination of Five Amphetamines in Human Hair,” Journal of Analytical Toxicology, Vol. 29, No. 2, 2005, pp. 135-139.
[12] R. Gottardo, F. Bortolotti, G. De Paoli, J. P. Pascali, I. Miksík and F. Tagliaro, “Hair Analysis for Illicit Drugs by Using Capillary Zone Electrophoresis-Electrospray Ionization-Ion Trap Mass Spectrometry,” Journal of Chromatography A, Vol. 1159, No. 1-2, 2007, pp. 185-189.
[13] S. Broecker, S. Herre and F. Pragst, “General Unknown Screening in Hair by Liquid Chromatography-Hybrid Quadrupole Time-of-Flight Mass Spectrometry (LC- QTOF-MS),” Forensic Science International, Vol. 218, No. 1, 2012, pp. 68-81.
[14] M. K. K. Nielsen, S. S. Johansen, P. W. Dalsgaard and K. Linnet, “Simultaneous Screening and Quantification of 52 Common Pharma-ceuticals and Drugs of Abuse in Hair Using UPLC-TOF-MS,” Forensic Science International, Vol. 196, No. 1-3, 2010, pp. 85-92. doi:10.1016/j.forsciint.2009.12.027
[15] S. Hegstad, H. Z. Khiabani, L. Kristoffersen, N. Kun?e, P. P. Lobmaier and A. S. Christophersen, “Drug Screening of Hair by Liquid Chromatography-Tandem Mass Spectrometry,” Journal of Analytical Toxicology, Vol. 32, No. 5, 2008, pp. 364-372.
[16] C. Hartmann, J. Smeyers-Verbeke, D. Massart and R. McDowall, “Validation of Bioanalytical Chromatographic Methods,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 17, No. 2, 1998, pp. 193-218. doi:10.1016/S0731-7085(97)00198-2
[17] F. T. Peters, O. H. Drummer and F. Musshoff, “Validation of New Methods,” Forensic Science International, Vol. 165, No. 2-3, 2007, pp. 216-224. doi:10.1016/j.forsciint.2006.05.021
[18] B. K. Matuszewski, M. L. Constanzer and C. M. Chavez- Eng, “Strategies for the Assessment of Matrix Effect in Quantitative Bioanalytical Methods Based on HPLC- MS/MS,” Analytical Chemistry, Vol. 75, No. 13, 2003, pp. 3019-3030. doi:10.1021/ac020361s
[19] A. Pelander, J. Ristimaa, I. Rasanen, E. Vuori and I. Ojanper?, “Screening for Basic Drugs in Hair of Drug Addicts by Liquid Chromatography/Time-of-Flight Mass Spectrometry,” Therapeutic Drug Monitoring, Vol. 30, No. 6, 2008, pp. 717-724. doi:10.1097/FTD.0b013e3181897cfa
[20] M. Villain, “Determination of Bromazepam, Clonazepam and Meta-bolites after a Single Intake in Urine and Hair by LC-MS/MS: Application to Forensic Cases of Drug Facilitated Crimes,” Forensic Science International, Vol. 145, No. 2-3, 2004, pp. 123-130.
[21] R. Stanaszek and W. Piekoszewski, “Simultaneous Determination of Eight Underivatized Amphetamines in Hair by High-Performance Liquid Chromatography-At- mospheric Pressure Chemical Ionization Mass Spectrometry (HPLC-APCI-MS),” Journal of Analytical Toxicology, Vol. 28, No. 2, 2004, pp. 77-85.
[22] R. Kronstrand, I. Nystr?m, J. Strandberg and H. Druid, “Screening for Drugs of Abuse in Hair with Ion Spray LC-MS-MS,” Forensic Science International, Vol. 145, No. 2-3, 2004, pp. 183-190. doi:10.1016/j.forsciint.2004.04.034

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