Abstract
A new quantitation method for the determination of drugs of abuse (opiates, amphetamine and derivatives, cocaine, methadone and metabolites) in serum by using online extraction coupled to liquid chromatography (LC)–mass spectrometry (MS)/MS has been developed. The online extraction is carried out using two extraction columns simultaneously and one analytical column. One extraction column is loaded, while the other one is eluted by a gradient. The elution gradient also separates the analytes in the analytical column. For the sample preparation, serum is spiked with a mixture of deuterated analogues of the drugs. After protein precipitation with methanol/zinc sulphate, centrifugation, evaporation and reconstitution, the sample is injected into the LC system. The quantitation is based on the analysis of two multiple reaction monitoring transitions per drug. The recovery of the protein precipitation step is over 80% for all analytes. Intra- and interday precision, as relative standard deviation, is lower than 6%, and in the case of accuracy, RE is lower than 15%. Only the most polar analytes showed matrix effects. The limits of quantitation for the analysed compounds vary between 0.5 and 2.8 ng/mL. The developed method was used to quantify basic drugs in samples “from driving under the influence of drugs” cases. The results were compared with those obtained by using solid-phase extraction–GC–MS.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alnouti Y, Srinivasan K, Waddell D, Bi H, Kavetskaia O, Gusev AI (2005) Development and application of a new on-line SPE system combined with LC–MS/MS detection for high throughput direct analysis of pharmaceutical compounds in plasma. J Chromatogr A 1080:99–106
Buszewski B, Welerowicz T, Tegowska E, Krzeminski TF (2009) Determination of selected beta-receptor antagonists in biological samples by solid phase extraction with cholesterolic phase and LC/MS. Anal Bioanal Chem 393:263–272
Jenkins KM, Young MS, Mallet CR, Elian AA (2004) Mixed-mode solid-phase extraction procedures for the determination of MDMA and metabolites in urine using LC–MS, LC–UV, or GC-NPD. J Anal Toxicol 28:50–58
Svensson JO, Andersson M, Gustavsson E, Beck O (2007) Electrospray LC–MS method with solid-phase extraction for accurate determination of morphine-, codeine-, and ethylmorphine-glucuronides and 6-acetylmorphine in urine. J Anal Toxicol 31:81–86
Herrin GL, McCurdy HH, Wall WH (2005) Investigation of an LC–MS–MS (QTrap) method for the rapid screening and identification of drugs in postmortem toxicology whole blood samples. J Anal Toxicol 29:599–606
Liu AC, Lin TY, Su LW, Fuh MR (2008) Online solid-phase extraction liquid chromatography–electrospray–tandem mass spectrometry analysis of buprenorphine and three metabolites in human urine. Talanta 75:198–204
Wu TY, Fuh MR (2005) Determination of amphetamine, methamphetamine, 3, 4-methylenedioxyamphetamine, 3, 4-methylenedioxyethylamphetamine, and 3, 4-methylenedioxymethamphetamine in urine by online solid-phase extraction and ion-pairing liquid chromatography with detection by electrospray tandem mass spectrometry. Rapid Commun Mass Spectrom 19:775–780
Kousoulosa C, Dotsikasa Y, Loukas YL (2007) Turbulent flow and ternary columnn-switching on-line clean-up system for high-throughput quantification of risperidone and its main metabolite in plasma by LC–MS/MS Application to a bioequivalence study. Talanta 72:360–367
Xia YQ, Whigan DB, Powell ML, Jemal M (2000) Ternary-column system for high-throughput direct-injection bioanalysis by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 14:105–111
Weinmann W, Renz M, Vogt S, Pollak S (2000) Automated solid-phase extraction and two-step derivatisation for simultaneous analysis of basic illicit drugs in serum by GC/MS. Int J Legal Med 113:229–235
Polson C, Sarkar P, Incledon B, Raguvaran V, Grant R (2003) Optimization of protein precipitation based upon effectiveness of protein removal and ionization effect in liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 785:263–275
Chambers E, Wagrowski-Diehl DM, Lu Z, Mazzeo JR (2007) Systematic and comprehensive strategy for reducing matrix effects in LC/MS/MS analyses. J Chromatogr B Analyt Technol Biomed Life Sci 852:22–34
Niessen WM (2003) Progress in liquid chromatography–mass spectrometry instrumentation and its impact on high-throughput screening. J Chromatogr A 1000:413–436
Matuszewski BK, Constanzer ML, Chavez-Eng CM (2003) Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC–MS/MS. Anal Chem 75:3019–3030
Peters FT, Hartung M, Herbold M, Schmitt G (2004) Anlage zu den Richtlinien der GTFCh zur Qualitätssicherung bei forensisch-toxikologischen Untersuchungen. Validierung. Toxichem + Krimtech 71:146–154
Schmidt GHM, Aderjan R (2004) B.E.N.: Bestimmungs-, Erfassungs-, und Nachweisgrenzen. Software Version 2.0
Acknowledgement
N. Ferreirós thanks the Basque Government for her postdoctoral grant.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ferreirós Bouzas, N., Dresen, S., Munz, B. et al. Determination of basic drugs of abuse in human serum by online extraction and LC–MS/MS. Anal Bioanal Chem 395, 2499–2507 (2009). https://doi.org/10.1007/s00216-009-3036-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-009-3036-x