Abstract
A method for sequential extraction of amphetamines (AMPs), opiates (OPIs), and 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) from a limited volume of urine was developed. The method used two different spin columns packed with octadecyl (C18) and mixed-mode C18-strong anion exchange (SAX)-bonded monolithic silica for extraction before analysis by gas chromatography–mass spectrometry (GC–MS). The urine (0.5 ml), which was hydrolyzed with 10 M NaOH solution (0.1 ml), and 0.3 M Sorensen’s glycine buffer (pH 13, 0.1 ml) were poured into the preactivated C18-SAX spin column and then centrifuged at 10,000 rpm for 2 min to load the sample solution. AMPs and THC-COOH, which adsorbed to the column, were eluted with methanol containing 2% HCOOH. For the second extraction, the remaining solution, which was hydrolyzed by acid, and 0.2 M Sorensen’s glycine buffer (pH 12, 0.1 ml) were poured into the other preactivated C18 spin column. The adsorbed OPIs were subsequently eluted with methanol containing 2% NH4OH. The eluates were combined and the solvent evaporated under nitrogen before the target drugs in the residue were successively converted to O-trimethylsilyl-N-trifluoroacetyl derivatives by N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA)-trimethylchlorosilane (TMCS) and N-methyl-bis(trifluoroacetamide) (MBTFA) followed analysis by GC–MS. Linearity from 10 to 1,000 ng/ml was observed for all the tested drugs using an internal standard method. The correlation coefficients of the calibration curves were greater than 0.990. The coefficients for intraday and interday variations at 50, 200, and 800 ng/ml in urine were between 0.7 and 11.1 %. The proposed method was applied to forensic and clinical poisoning cases and is considered suitable for routine analysis of drugs in biological materials.
Similar content being viewed by others
References
McDowall RD (1989) Sample preparation for biomedical analysis. J Chromatogr B 492:3–58
Chang MS, Ji Q, Zhang J, El-Shourbagy TA (2007) Historical review of sample preparation for chromatographic bioanalysis: pros and cons. Drug Develop Res 68:107–133
Kole PL, Venkatesh G, Kotecha J, Sheshala R (2011) Recent advances in sample preparation techniques for effective bioanalytical methods. Biomed Chromatogr 25:199–217
Namera A, Nakamoto A, Saito T, Nagao M (2011) Colorimetric detection and chromatographic analyses of designer drugs in biological materials: a comprehensive review. Forensic Toxicol 29:1–24
El-Haj BM, Ali HS, Hamoudi NM (2011) Oripavine as a new marker of opiate product use. Forensic Toxicol 29:152–158
Murphy MA, Huestis MA (2005) LC-ESI-MS/MS analysis for the quantification of morphine, codeine, morphine-3-β-D-glucuronide, morphine-6-β-D-glucuronide, and codeine-6-β-D-glucuronide in human urine. J Mass Spectrom 40:1412–1416
Shima N, Katagi M, Kamata H, Zaitsu K, Kamata T, Miki A, Tsuchihashi H, Sakuma T, Nemoto N (2008) Conjugates of p-hydroxymethamphetamine and 4-hydroxy-3-methoxymethamphetamine in blood obtained from methamphetamine and 3,4-methylenedioxymethamphetamine users: analysis by LC-MS-MS. Forensic Toxicol 26:58–65
Kłys M, Rojek S (2008) Four nonfatal and six fatal cases of opiate use: utility of morphine, its metabolites, and their ratios in blood specimens. Forensic Toxicol 26:87–90
Felli M, Martello S, Chiarotti M (2011) 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
Deventer K, Pozo OJ, Delbeke FT, Van Eenoo P (2012) Direct quantification of morphine glucuronides and free morphine in urine by liquid chromatography–tandem mass spectrometry. Forensic Toxicol 30:106–113
Namera A, Nakamoto A, Nishida M, Saito T, Kishiyama I, Miyazaki S, Yahata M, Yashiki M, Nagao M (2008) Extraction of amphetamines and methylenedioxyamphetamines from urine using a monolithic silica disk-packed spin column and high-performance liquid chromatography-diode array detection. J Chromatogr A 1208:71–75
Saito T, Yamamoto R, Inoue S, Kishiyama I, Miyazaki S, Nakamoto A, Nishida M, Namera A, Inokuchi S (2008) Simultaneous determination of amitraz and its metabolite in human serum by monolithic silica spin column extraction and liquid chromatography–mass spectrometry. J Chromatogr B 867:99–104
Saito T, Miura N, Namera A, Oikawa H, Miyazaki S, Nakamoto A, Inokuchi S (2012) Mixed-mode C-C18 monolithic spin-column extraction and GC–MS for simultaneous assay of organophosphorus compounds, glyphosate, and glufosinate in human serum and urine. Forensic Toxicol 30:1–10
Nakanishi K (1997) Pore structure control of silica gels based on phase separation. J Porous Mater 4:67–112
Miyazaki S, Yamada T, Ohta S, Saito T, Namera A, Ohira M (2010) Rapid preparation methods of biological samples for ionic compounds using ion exchange type monolithic silica spin column (in Japanese). Bunseki Kagaku 59:213–218
Delbeke FT, Debackere M (1993) Influence of hydrolysis procedures on the urinary concentrations of codeine and morphine in relation to doping analysis. J Pharm Biomed Anal 11:339–343
Chen ZR, Somogyi AA, Reynolds G, Bochner F (1991) Disposition and metabolism of codeine after single and chronic doses in one poor and seven extensive metabolisers. Br J Clin Pharmacol 31:381–390
Harvey DJ, Paton WDM (1984) Metabolism of the cannabinoids. Rev Biochem Toxicol 6:221–264
Kelly P, Jones RT (1992) Metabolism of tetrahydrocannabinol in frequent and infrequent marijuana users. J Anal Toxicol 16:228–235
Raharjo TJ, Verpoorte R (2004) Methods for the analysis of cannabinoids in biological materials: a review. Phytochem Anal 15:79–94
Williams PL, Moffat AC (1980) Identification in human urine of Δ9-tetrahydrocannabinol-11-oic acid glucuronide: a tetrahydrocannabinol metabolite. J Pharm Pharmacol 32:445–448
Kemp PM, Abukhalaf IK, Manno JE, Manno BR, Alford DD, McWilliams ME, Nixon FE, Fitzgerald MJ, Reeves RR, Wood MJ (1995) Cannabinoids in humans. II. The influence of three methods of hydrolysis on the concentration of THC and two metabolites in urine. J Anal Toxicol 19:292–298
Kramer E, Kovar KA (1999) On-line coupling of automated solid-phase extraction with high-performance liquid chromatography and electrochemical detection: quantitation of oxidizable drugs of abuse and their metabolites in plasma and urine. J Chromatogr 731:167–177
Abraham TT, Lowe RH, Pirnay SO, Darwin WD, Huestis MA (2007) Simultaneous GC-EI-MS determination of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol, and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in human urine following tandem enzyme-alkaline hydrolysis. J Anal Toxicol 31:477–485
Ranieri TL, Ciolino LA (2008) Rapid selective screening and determination of ephedrine alkaloids using GC–MS footnote mark. Phytochem Anal 19:127–135
Guidance for Industry Bioanalytical Method Validation. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm070107.pdf. Accessed 6 December 2012
Namera A, Yamamoto S, Saito T, Miyazaki S, Oikawa H, Nakamoto A, Nagao M (2011) Simultaneous extraction of acidic and basic drugs from urine using mixed-mode monolithic silica spin column bonded with octadecyl and cation-exchange group. J Sep Sci 34:2232–2239
Namera A, Nakamoto A, Miyazaki S, Yui Y, Saito T, Nagao M (2011) Monolithic spin column: a new extraction device for analysis of drugs in urine and serum by GC–MS and HPLC–MS. J AOAC Int 94:765–774
Valente MJ, Carvalho F, Bastos ML, Carvalho M, De Pinho PG (2010) Development and validation of a gas chromatography/ion trap-mass spectrometry method for simultaneous quantification of cocaine and its metabolites benzoylecgonine and norcocaine: application to the study of cocaine metabolism in human primary cultured renal cells. J Chromatogr B 878:3083–3088
Ellison ST, Brewer WE, Morgan SL (2009) Comprehensive analysis of drugs of abuse in urine using disposable pipette extraction. J Anal Toxicol 33:356–365
Acknowledgments
The authors thank Dr. K. Hara of Fukuoka University (Fukuoka, Japan) for donating the standard compound (pentadeuterated MA hydrochloride).
Conflict of interest
There is no conflict of interest to report.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Namera, A., Saito, T., Miyazaki, S. et al. Sequential extraction of amphetamines, opiates, and 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid from a limited volume of urine using a monolithic silica spin column coupled with gas chromatography–mass spectrometry. Forensic Toxicol 31, 312–321 (2013). https://doi.org/10.1007/s11419-013-0185-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11419-013-0185-6