Development and validation of a qualitative GC-MS method for methamphetamine and amphetamine in human urine using aqueous-phase ethyl chloroformate derivatization
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Methamphetamine (MA) is the most common and available drug of abuse in Korea and its primary metabolite is amphetamine (AP). Detection of AP derivatives, such as MA, AP, phentermine (PT), MDA, MDMA, and MDEA by the use of immunoassay screening is not reliable and accurate due to cross-reactivity and insufficient specificity/sensitivity. Therefore, the analytical process accepted by most urine drug-testing programs employs the two-step method with an initial screening test followed by a more specific confirmatory test if the specimen screens positive. In this study, a gas chromatography-mass spectrometric (GC-MS) method was developed and validated for confirmation of MA and AP in human urine. Urine sample (500 μL) was added with N-isopropylbenzylamine as internal standard and ethyl chloroformate as a derivatization reagent, and then extracted with 200 μL of ethyl acetate. Extracted samples were analysed with GC-MS in the SIM/ Scan mode, which were screened by Cobas c311 analyzer (Roche/Hitachi) to evaluate the efficiency as well as the compatibility of the GC-MS method. Qualitative method validation requirements for selectivity, limit of detection (LOD), precision, accuracy, and specificity/sensitivity were examined. These parameters were estimated on the basis of the most intense and characteristic ions in mass spectra of target compounds. Precision and accuracy were less than 5.2 % (RSD) and ±14.0 % (bias), respectively. The LODs were 3 ng/mL for MA and 1.5 ng/mL for AP. At the screening immunoassay had a sensitivity of 100% and a specificity of 95.1 % versus GC-MS for confirmatory testing. The applicability of the method was tested by the analysis of spiked urine and abusers' urine samples.
Abstract
Methamphetamine (MA) is the most common and available drug of abuse in Korea and its primary metabolite is amphetamine (AP). Detection of AP derivatives, such as MA, AP, phentermine (PT), MDA, MDMA, and MDEA by the use of immunoassay screening is not reliable and accurate due to cross-reactivity and insufficient specificity/sensitivity. Therefore, the analytical process accepted by most urine drug-testing programs employs the two-step method with an initial screening test followed by a more specific confirmatory test if the specimen screens positive. In this study, a gas chromatography-mass spectrometric (GC-MS) method was developed and validated for confirmation of MA and AP in human urine. Urine sample (500 μL) was added with N-isopropylbenzylamine as internal standard and ethyl chloroformate as a derivatization reagent, and then extracted with 200 μL of ethyl acetate. Extracted samples were analysed with GC-MS in the SIM/ Scan mode, which were screened by Cobas c311 analyzer (Roche/Hitachi) to evaluate the efficiency as well as the compatibility of the GC-MS method. Qualitative method validation requirements for selectivity, limit of detection (LOD), precision, accuracy, and specificity/sensitivity were examined. These parameters were estimated on the basis of the most intense and characteristic ions in mass spectra of target compounds. Precision and accuracy were less than 5.2 % (RSD) and ±14.0 % (bias), respectively. The LODs were 3 ng/mL for MA and 1.5 ng/mL for AP. At the screening immunoassay had a sensitivity of 100% and a specificity of 95.1 % versus GC-MS for confirmatory testing. The applicability of the method was tested by the analysis of spiked urine and abusers' urine samples.
참고문헌 (31)
- 김소연, "약물 오남용의 신경생물학적 기전과 약동학 및 약력학", 대한의사협회지, 56(9), pp.762~770 (2013), 대한의사협회
- 문유호, "메스암페타민(필로폰) 급성 중독으로 유발된 심정지 후 생존한 1례", 대한임상독성학회지, 16(2), pp.176~180 (2018), 대한임상독성학회[DOI]
- M. D. Anglin, , J. Psychoactive Drugs, 32, pp.137~141 (2000)
- T. K. Mackey, , J. Med. Internet Res, 15, pp.665~689 (2013)
- J. Lee, , Forensic Sci. Int, 272, pp.1~9 (2017)
- White Paper on Drug-Related Crimes 2018, "Supreme Prosecutors' Office", ~ (2019)
- N. J. Kwon, , Forensic Sci. Int, 286, pp.81~85 (2018)
- J. Caldwell, , Biochem. J, 129, pp.11~22 (1972)
- Y. Xue, , Biochem. Biophys. Res. Commun, 509, pp.395~401 (2019)
- J. R. Richards, , Drug Alcohol Depend, 150, pp.1~13 (2015)
- G. M. Reisfield, , Bioanalysis, 1, pp.937~952 (2009)
- A. Saitman, , J. Anal. Toxicol, 38, pp.387~396 (2014)
- A. D. de Jager, , J. Chromatogr. B, 879, pp.2642~2652 (2011)
- S. J. Mulé, , J. Anal. Toxicol, 12, pp.102~107 (1988)
- E. R. Perez, , J. Anal. Toxicol, 40, pp.201~207 (2016)
- D. K. Lee, , Food Chem, 141, pp.3931~3937 (2013)
- J. M. Halket, , J. Exp. Bot, 56, pp.219~243 (2005)
- C. Jiménez, , J. Chromatogr. B, 767, pp.341~351 (2002)
- E. Trullols, , Trends Anal. Chem, 23, pp.137~145 (2004)
- R. Fogerson, , J. Anal. Toxicol, 21, pp.451~458 (1997)
- P. S. Cheng, , J. Chromatogr. B, 852, pp.443~449 (2007)
- M. Gaugain-Juhel, , Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess, 26, pp.1459~1471 (2009)
- P. Hušek, , J. Chromatogr. B, 717, pp.57~91 (1998)
- K. Mohamed, , J. Anal. Toxicol, 41, pp.639~645 (2017)
- J. -Y. Moon, , J. Lipid Res, 52, pp.1595~1603 (2011)
- Y. Qiu, , Anal. Chim. Acta, 583, pp.277~283 (2007)
- S. P. Elliott, , Sci. Justice, 58, pp.335~345 (2018)
- A. G. Verstraete, , J. Anal. Toxicol, 29, pp.359~364 (2005)
- A. J. Barnes, , J. Anal. Toxicol, 27, pp.402~407 (2003)
- W. Y. Pyo, , Chromatographia, 64, pp.731~737 (2006)
- 정재철, "고체상추출과 GC", MS를 이용한 소변 중 암페타민계 마약성분 동시분석법, 2008, pp.21(1), 41~47[DOI]