title: Determination of co-administrated opioids and benzodiazepines in urine using column-switching solid-phase extraction and liquid chromatography-tandem mass spectrometry.
abstract: Co-administration of opioids with benzodiazepines is very common around the world. A semi-automated method was developed for the determination of four opioids and two benzodiazepines as well as their metabolites (including glucuronide metabolites) in human urine, based on on-line column-switching-solid-phase extraction (CS-SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The CS-SPE was performed by loading 200μL of urine sample to an Oasis HLB cartridge. Detection was achieved using a LC-MS/MS system equipped with an electrospray ionization source (ESI). For unequivocal identification and confirmation, two selected reaction monitoring transitions were registered for each compound, and no co-elution of interferences was observed at the expected retention time. Significant ion suppressions were observed for most analytes during chromatographic runs, but isotope-labeled internal standards (ISs) were used and found to be useful to compensate for the determination error caused by the matrix effect. The assay's linearity ranged from 1-20ng/mL to 800-1000ng/mL for 23 compounds, except for lorazepam (LOR), whose linearity was in the range of 1-100ng/mL. This method showed to be precise and accurate. The relative standard deviation (RSD) % values of within-run precision, between-run precision and total precision were not greater than 10.4% (n=3), 12.9% (n=5) and 15.1% (n=15), respectively. Accuracy values were in the range of 87.5-110%. Limits of detection (LODs) ranged from 0.2ng/mL to 5ng/mL, and limits of quantification (LOQs) ranged from 1ng/mL to 20ng/mL. The method was applied to the assay of 12 samples from forensic cases, which exemplified the co-administration of benzodiazepines (BZDs) by some heroin abusers. This method was of high sensitivity, selectivity and reliability, minimum sample manipulation, semi-automation, and fairly high throughput (analysis time per sample was 20min). The method developed will be useful for the detection of co-administrated drugs and the study of the interactions of BDZs with opioids.
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China; State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, PR China; Pharmacy Department of Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, PR China.
authors.forename: Lingjuan
authors.initials: L
authors.lastname: Xiong
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China.
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China.
authors.forename: Chen
authors.initials: C
authors.lastname: Liang
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China; State Key Laboratory of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, Shanghai, PR China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Teng
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China; Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, PR China.
authors.forename: Fengli
authors.initials: F
authors.lastname: Jiang
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China.
authors.forename: Libo
authors.initials: L
authors.lastname: Zeng
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Ye
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China.
authors.forename: Chunfang
authors.initials: C
authors.lastname: Ni
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China.
authors.forename: Xiaoliang
authors.initials: X
authors.lastname: Yuan
authors.affiliation: Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, PR China. Electronic address: yulan_rao@fudan.edu.cn.
authors.forename: Yulan
authors.initials: Y
authors.lastname: Rao
authors.affiliation: Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Institute of Forensic Science, Shanghai, PR China. Electronic address: yr_zhang@126.com.
authors.forename: Yurong
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000701:Analgesics, Opioid / Q000652:urine*; D001569:Benzodiazepines / Q000652:urine*; D002853:Chromatography, Liquid; D020719:Glucuronides; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry; D016482:Urinalysis / Q000379:methods*
chemical_list: D000701:Analgesics, Opioid; D020719:Glucuronides; D001569:Benzodiazepines
keywords: Benzodiazepines; Co-administration; Column-switching solid-phase extraction; Glucuronide metabolites; Opioids; Urine