Abstract
The advent of LC combined with MS made it possible to design analytical methods for urine drug testing based on the very simple concept of diluting urine with an internal standard as the sole preparation procedure prior to instrumental analysis. The number of publications using this method design increased after the development of high-efficiency LC based on sub-2 μm particles. The success of this method design for drug testing, doping control and toxicological investigations of urine is now well documented and comprise both screening and confirmation methods. The nondiscriminating nature of this method design makes it even more attractive in combination with high-resolution MS for multicomponent target and general unknown analysis applications.
Papers of special note have been highlighted as: • of interest
References
- 1 . Clinical issues associated with urine testing of substances of abuse. Pharmacotherapy 17, 497–510 (1997).
- 2 . Analytical and technical aspects of testing for drug abuse: confirmatory procedures. Clin. Chem. 34, 471–473 (1988).
- 3 . Urine testing in the detection of drugs of abuse. Arch. Intern. Med. 148, 2407–2412 (1988).
- 4 . Designer drugs: a medicinal chemistry perspective. Ann. NY Acad. Sci. 48, 18–38 (2012).• Overview of the rapid growing new psychoactives substances.
- 5 . New developments in biochemical mass spectrometry: electrospray ionization. Anal. Chem. 62, 882–899 (1990).
- 6 . High-performance liquid chromatography/mass spectrometry: state of the art for the drug analysis laboratory. Clin. Chem. 35, 1282–1287 (1989).
- 7 . Recent advances in high-throughput quantitative bioanalysis by LC–MS/MS. J. Pharm. Biomed. Anal. 44, 342–355 (2007).
- 8 . Determination of drugs in biological fluids by direct injection of samples for liquid-chromatographic analysis. J. Biochem. Biophys. Methods 70, 263–273 (2007).• Overview of different solutions for direct injection of biofluids.
- 9 . Introduction of solid-phase microextraction as a high-throughput sample preparation toll in laboratory analysis of prohibited substances. Anal. Chim. Acta 809, 69–81 (2014).
- 10 . High-throughput bioanalytical method for analysis of synthetic cannabinoid metabolites in urine using salting-out sample preparation and LC–MS/MS. J. Chromatogr. B 909, 42–50 (2012).
- 11 . Direct tandem mass spectrometry for the simultaneous assay of opioids cocoaine, and metabolites in dried urine spots. Anal. Chim. Acta 784, 25–32 (2013).
- 12 . Drugs of abuse screening in urine as part of a metabolite-based LC–MSn screening concept. Anal. Bioanal. Chem. 400, 3481–3489 (2011).
- 13 . Dependence of ion intensity in electrospray mass spectrometry on the concentration of the analytes in the electrosprayed solution. Anal. Chem. 65, 3654–3668 (1993).
- 14 . Signal suppression/enhancement in high-performance liquid chromatography tandem mass spectrometry. J. Chromatogr. A 1217, 3929–3937 (2010).• Summary of knowledge regarding matrix effects on ionization.
- 15 . Validation of bioanalytical LC–MS/MS assays: evaluation of matrix effects. J. Chromatogr. B 877, 2198–2207 (2009).
- 16 . Aspects of matrix effects in applications of liquid chromatography-mass spectrometry to forensic and clinical toxicology – a review. Anal. Bioanal. Chem. 403, 2155–2172 (2012).
- 17 . Mechanistic investigation of ionization suppression in electrospray ionization. J. Am. Soc. Mass Spectrom. 11, 942–950 (2000).• Important article evaluating matrix effects.
- 18 . Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC–MS/MS. Anal. Chem. 75, 3019–3030 (2003).• Important article evaluating matrix effects.
- 19 The human urine metabolome. PLoS One 8(9), e73076 (2013).
- 20 . Standard line slopes as a measure of a relative matrix effect in quantitative HPLC–MS bioanalysis. J. Chromatogr. B 830, 293–300 (2006).
- 21 . Matrix effect in bio-analysis of illicit drugs with LC–MS/MS: influence of ionization type, sample preparation, and biofluid. J. Am. Soc. Mass Spectrom. 14, 1290–1294 (2003).
- 22 Simultaneous analysis of gamma-hydroxybutyric acid and its precursors in urine using liquid chromatography-tandem mass spectrometry. J. Chromatogr. A 1056, 83–90 (2004).
- 23 . Validation of direct injection electrospray LC–MS/MS for confirmation of opiates in urine drug testing. J. Mass Spectrom. 42, 881–889 (2007).
- 24 . Direct injection LC–MS/MS method for identification and quantification of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylenedioxymethamphetamine in urine drug testing. J. Chromatog. B 861, 22–28 (2008).
- 25 . Matrix effects: the Achilles heel of quantitative high-performance liquid chromatography-electrospray-tandem mass spectrometry. Clin. Biochem. 38, 328–334 (2005).
- 26 . Progress in liquid chromatography-mass spectrometry instrumentation and its impact on high-throughput screening. J. Chromatogr. A 1000, 413–436 (2003).
- 27 . Liquid chromatography-mass spectrometry in forensic and clinical toxicology. J. Chromatogr. B 713, 3–25 (1998).• This author has repeatedly reviewed the field and provided regular updates.
- 28 . Sample preparation for LC/MS/MS: knowing the basic requirements and the bid picture of an LC/MS system can ensure success in most instances. Anal. Chem. 70, 650A–656A (1998).
- 29 . Rapid confirmation/quantitation of cocaine and benzoylecgonine in urine utilizing high-performance liquid chromatography and tandem mass spectrometry. J. Am. Soc. Mass Spectrom. 11, 247–263 (2000).
- 30 . Rapid confirmation/quantitation of ecgonine methyl ester, benzoylecgonine and cocaine in urine using on-line extraction coupled with fast HPLC and tandem mass spectrometry. J. Anal. Toxicol. 25, 69–75 (2001).
- 31 . Broad spectrum drug identification directly from urine, using liquid chromatography-tandem mass spectrometry. Clin. Chem. 45, 1224–1234 (1999).
- 32 . Urine drug testing for opioids, cocaine, and metabolites by direct injection liquid chromatography/tandem mass spectrometry. Rapid Comm. Mass Spectrom. 17, 1665–1670 (2003).
- 33 . Direct screening of urine for MDMA and MDA by liquid chromatography-tandem mass spectrometry. J. Anal. Toxicol. 27, 15–19 (2003).
- 34 . Analysis of buprenorphine, nor buprenorphine, and their glucuronides in urine by liquid chromatography-mass spectrometry. J. Anal. Toxicol. 27, 464–470 (2003).
- 35 . Urinary screening for methylphenidate (Ritalin) abuse: a comparison of liquid chromatography-tandem mass spectrometry, gas chromatography-mass spectrometry, and immunoassay methods. Clin. Biochem. 37, 175–183 (2004).
- 36 . Ethyl glucuronide – a marker of alcohol consumption and a relapse marker with clinical and forensic implications. Alcohol Alcohol. 34, 71–77 (1999).
- 37 . Direct quantification of ethyl glucuronide in clinical urine samples by liquid chromatography-mass spectrometry. Ther. Drug Monitor. 24, 645–651 (2002).
- 38 . Confirmatory analysis of ethylglucuronide in urine by liquid-chromatography/electrospray ionization/tandem mass spectrometry according to forensic guidelines. J. Am. Soc. Mass Spectrom. 15, 188–193 (2004).
- 39 . Comparison of analytical approaches for liquid chromatography/mass spectrometry determination of the alcohol biomarker ethyl glucuronide in urine. Rapid Comm. Mass Spectrom. 24, 1737–1743 (2010).
- 40 . Validated method for the determination of the ethanol consumption markers ethyl glucuronide, ethyl phosphate, and ethyl sulfate in human urine by reversed-phase/weak anion exchange liquid chromatography-tandem mass spectrometry. Anal. Chem. 78, 5884–5892 (2006).
- 41 . Multicomponent screening for drugs of abuse. Direct analysis of urine by LC–MS–MS. Ther. Drug Monitor. 26, 90–97 (2004).
- 42 . Application of direct urine LC–MS–MS analysis screening of novel substances in drug abusers. J. Anal. Toxicol. 29, 234–238 (2005).
- 43 . Development and clinical application of an LC–MS–MS method for mescaline in urine. J. Anal. Toxicol. 32, 227–231 (2008).
- 44 . A multi-component LC–MS/MS method for detection of ten plant-derived psychoactive substances in urine. J. Chromatogr. B 877, 1162–1169 (2009).
- 45 . A high-throughput multicomponent screening method for diuretics, masking agents, central nervous system (CNS) stimulants and opiates in human urine by UPLC–MS/MS. J. Mass Spectrom. 43, 980–992 (2008).
- 46 . Multicomponent LC–MS/MS screening method for detection of new psychoactive drugs, legal highs, in urine – experience from the Swedish population. J. Chromatogr. B 930, 112–120 (2013).
- 47 . Identification of novel psychoactive drug use in Sweden based on laboratory analysis – initial experiences from the STRIDA project. Scand. J. Clin. Lab. Invest. 73, 400–406 (2013).
- 48 . Detection of new psychoactive substance use among emergency room patients: Results from the Swedish STRIDA project. Forensic Sci. Int. 243C, 23–29 (2014).
- 49 . Simultaneous determination of γ-hydroxybutyrate (GHB) and its analogues (GBL, 1.4-BD, GVL) in whole blood and urine by liquid chromatography coupled to tandem mass spectrometry. J. Anal. Toxicol. 35, 8–14 (2011).
- 50 . Determination of gamma-hydroxybutyrate (GHB), beta-hydroxybutyrate (BHB), pregabalin, 1,4 butane-diol (1,4 BD) and gamma-butyrolactone (GBL) in whole blood and urine samples by UPLC–MSMS. J. Chromatogr. B 885–886, 37–42 (2012).
- 51 . Screening and confirmation methods for GHB determination in biological fluids. Ann. Bioanal. Chem. 406, 3553–3577 (2014).
- 52 . Development of a rapid LC–MS/MS method for direct urinalysis of designer drugs. Drug Test. Anal. 3, 496–504 (2011).
- 53 . Performance evaluation of three liquid chromatography mass spectrometry methods for broad spectrum drug screening. Clin. Chim. Acta 411, 1474–1481 (2010).
- 54 . Detection and identification of 700 drugs by multi-target screening with a 3200 Q TRAP® LC–MS/MS system and library searching. Anal. Bioanal. Chem. 396, 2425–2434 (2010).
- 55 . Simultaneous screening of 177 drugs of abuse in urine using ultra-performance liquid chromatography with tandem mass spectrometry in drug-intoxicated patients. Clin. Psychopharm. Neurosci. 11, 158–164 (2013).
- 56 . Broad spectrum drug screening using liquid chromatography-hybrid triple quadropole linear ion trap mass spectrometry. In: Clinical Applications of Mass Spectrometry. Garg U, Hammett-Stabler CA (Eds. Humana Press, USA, 203–218 (2010).
- 57 . Drugs of abuse testing by tandem mass spectrometry: a rapid, simple method to replace immunoassays. Clin. Biochem. 42, 1531–1542 (2009).• Demonstrates for the first time that LC–MS technology can replace immunoassay screening.
- 58 . Evaluation of a direct high-capacity target screening approach for urine drug testing using liquid chromatography-time-of-flight mass spectrometry. J. Chromatogr. B 909, 6–13 (2012).
- 59 . Screening and confirmation of 62 drugs of abuse and metabolites in urine by ultra-high-performance liquid chromatography - quadrupole time-of-flight mass spectrometry. J. Anal. Toxicol. 37, 642–651 (2013).
- 60 . Evaluation of high-resolution mass spectrometry for urine toxicology screening in a pain management setting. J. Anal. Toxicol. 36, 601–607 (2012).
- 61 . Confirmatory analysis for drugs of abuse in plasma and urine by high-performance liquid chromatography–tandem mass spectrometry with respect to criteria for compound identification. J. Chromatog. B 811, 21–30 (2004).
- 62 European Union Decision 2002/657/EC. http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2002:221:0008:0036:ES:PDF.
- 63 US Department of Health and Human Services, Food and Drug Administration (FDA). Center for Veterinary Medicine, 1 May. Guidance for Industry. Mass Spectrometry for Confirmation of the Identity of Animal Drug Residues (FDA Guidance Document 118). www.fda.gov/.../AnimalVeterinary/GuidanceComplianceEnforcement/ GuidanceforIndustry/ucm052658.pdf.
- 64 World Anti-Doping Agency. WADA Technical Document TD2003IDCR. http://www.wada-ama.org/rtecontent/document/criteria12.pdf.
- 65 . Dilute and shoot: analysis of drugs of abuse using selected reaction monitoring for quantification and full scan product ion spectra for identification. J. Anal. Toxicol. 36, 106–111 (2012).
- 66 . Accurate identification and quantification of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid in urine drug testing: evaluation of a direct high efficiency liquid chromatographic-mass spectrometric method. J. Chromatog. B 871, 101–108 (2008).
- 67 . Direct quantification of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid in urine by liquid chromatography/tandem mass spectrometry in relation to doping control analysis. Rapid Comm. Mass Spectrom. 24, 1133–1141 (2010).
- 68 . LC–MS–MS method for simultaneous determination of THCCOOH and THCCOOH-glucuronide in urine: application to workplace confirmation tests. Forensic Sci. Int. 204, 67–73 (2011).
- 69 Direct urine analysis for the identification and quantification of selected benzodiazepines for toxicology screening. J Chromatog. B 902, 42–46 (2012).
- 70 . Direct quantification of morphine glucuronides and free morphine in urine by liquid chromatography-tandem mass spectrometry. Forensic Toxicol. 30, 106–113 (2012).
- 71 . Direct and efficient liquid chromatographic tandem mass spectrometric method for opiates in urine drug testing – importance of 6-acetylmporphine and reduction of analytes. Drug Test. Anal. 6(4), 317–324 (2014).
- 72 . Direct analysis of opiates in urine by liquid chromatography-tandem mass spectrometry. J. Anal. Toxicol. 29, 704–710 (2005).
- 73 . Dilute and shoot-liquid chromatography-mass spectrometry for urine analysis in doping control and analytical toxicology. Trends Analyt. Chem. 55, 1–13 (2014).• Excellent review of the field.
- 74 WADA. Prohibited List 2014. World Anti-Doping Agency (WADA), Montreal, Canada (2013).
- 75 WADA. Technical Document Minimum Required Performance Limits for Detect ion of Prohibited Substances: TD2013MRPL. World Anti-Doping Agency (WADA), Montreal, Canada (2013).
- 76 High-throughput screening for various classes of doping agents using a new ‘dilute-and-shoot’ liquid chromatography-tandem mass spectrometry multi-target approach. Drug Test. Anal. 3, 836–850 (2011).
- 77 . Qualitative detection of diuretics and acidic metabolites of other doping agents in human urine by high-performance liquid chromatography–tandem mass spectrometry. Comparison between liquid–liquid extraction and direct injection. J. Chromatog. A 1216, 5819–5827 (2009).
- 78 . A direct screening procedure for diuretics in human urine by liquid chromatography-tandem mass spectrometry with information dependent acquisition. Clin. Chim. Acta 386, 46–52 (2007).
- 79 . A one-year monitoring of nicotine use in sport: Frontier between potential performance enhancement and addiction issues. Forensic Sci. Int. 213 73–84 (2011).
- 80 . Development and validation of an LC–MS/MS method for the quantification of ephedrines in urine. J. Chromatog. B 877, 369–374 (2009).
- 81 . A simple high pH liquid chromatography-tandem mass spectrometry method for basic compounds: application to ephedrines in doping control. J. Chromatog. A 1218, 2098–2105 (2011).
- 82 Doping control analysis in human urine by liquid chromatography–electrospray ionization ion trap mass spectrometry for the Olympic Games Athens 2004: determination of corticosteroids and quantification of ephedrines, salbutamol and morphine. Anal. Chim. Acta 573–574, 242–249 (2006).
- 83 Fast analysis of doping agents in urine by ultra-high-pressure liquid chromatography–quadrupole time-of-flight mass spectrometry I. Screening analysis.. J. Chromatog. A 1216, 4423–4433 (2009).
- 84 . Development and validation of an open screening method for diuretics, stimulants and selected compounds in human urine by UHPLC–HRMS for doping control. Anal. Chim. Acta 721, 137–146 (2012).
- 85 . Development of a universal metabolome-standard method for long-term LC–MS metabolome profiling and its application for bladder cancer urine-metabolite-biomarker discovery. Anal. Chem. 86(13), 6540–6547 (2014).
- 86 . Stable isotopically labeled internal standards in quantitative bioanalysis using liquid chromatography/mass spectrometry: necessity or not? Rapid Commun. Mass Spectrom. 19, 401–407 (2005).
- 87 . Does a stable isotopically labeled internal standard always correct analyte response? A matrix effect study on a LC/MS/MS method for the determination of carvedilol enantiomers in human plasma. J. Pharm. Biomed. Anal. 43, 701–707 (2007).
- 88 . 13C labelled internal standards – a solution to minimize ion suppression effects in liquid chromatography-tandem mass spectrometry analyses of drugs in biological samples? J. Chromatogr. A 1218, 9366–9374 (2011).
- 89 . Evaluation of 13C- and 2H-labeled internal standards for the determination of amphetamines in biological samples, by reversed-phase ultra-high performance liquid chromatography-tandem mass spectrometry. J. Chromatogr. A 1344, 83–90 (2014).