title: Rapid detection and quantification of 35 benzodiazepines in urine by GC-TOF-MS.
abstract: A rapid and sensitive method for the screening and quantification of 35 benzodiazepines in human urine by gas chromatography/time-of-flight mass spectrometry was developed and validated. Target analytes were isolated from 1 ml urine by solid-phase extraction using Oasis MCX extraction columns (extraction recovery between 35 and 99%). With a supported liquid-liquid extraction method, a new modification of conventional liquid-liquid-extraction, a less time intensive alternative for benzodiazepine extraction is presented. The sample pretreatment entails the derivatization of the benzodiazepines with N,O-bis(trimethylsilyl)trifluoroacetamide plus 1% trimethylchlorosilane. Separation of all benzodiazepines was done within 9.5 min, and detection was based on full mass spectra for each analyte. A deconvolution algorithm was used for unresolved chromatographic peaks to identify coeluted substances. The subsequent quantification was done using significant masses. The limit of quantification is 10 ng/ml for most of the compounds. Linearity is in the range between 10 and 350 ng/ml. Reproducibility was observed with coefficients of variation below 2% at concentrations of 50 and 200 ng/ml. The accuracy is between 88 and 108% depending on the respective analyte and the concentration.
authors.affiliation: Roche Diagnostics GmbH, Nonnenwald 2, 82377, Penzberg, Germany. kathrin.arnhard@contractors.roche.com
authors.forename: Kathrin
authors.initials: K
authors.lastname: Arnhard
authors.forename: Rupert
authors.initials: R
authors.lastname: Schmid
authors.forename: Uwe
authors.initials: U
authors.lastname: Kobold
authors.forename: Roland
authors.initials: R
authors.lastname: Thiele
mesh_terms: D000081:Acetamides / Q000737:chemistry; D001569:Benzodiazepines / Q000302:isolation & purification / Q000652:urine*; D008401:Gas Chromatography-Mass Spectrometry / Q000191:economics / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000379:methods; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods; D014297:Trimethylsilyl Compounds / Q000737:chemistry
chemical_list: D000081:Acetamides; C103255:N,O-bis(trimethylsilyl)trifluoroacetamide; D014297:Trimethylsilyl Compounds; D001569:Benzodiazepines; C039293:trimethylchlorosilane