Development of the MAE/UHPLC-MS-TOF method for determination of benzodiazepines in human bio-fluids for toxicological analysis.
A rapid method of microwave-assisted extraction (MAE) followed by ultrahigh performance liquid chromatography with mass spectrometry with time of flight detection (UHPLC-MS-TOF) was optimized and validated for the purpose of determination of five benzodiazepines in human serum and blood samples. Extraction parameters and conditions of the UHPLC-MS-TOF method were defined. Validation of the developed method was performed at three concentration levels: 10, 100 and 250 ng/mL of each drug for both serum and blood samples. For serum and blood the limit of detection was found in the ranges 0.46-2.58 ng/mL and 0.43-1.87 ng/mL, precision (RSD): 0.3-6.7% and 0.9-8.4%, accuracy of the assay (RE): -5.3 to +2.4% and -5.7 to +7.6%, recovery: 80.5-104.3% and 79.9-106.9%, matrix effects: 95.9-110.5% and 97.5-114.2%, respectively. Moreover, the optimized and validated MAE/UHPLC-MS-TOF method was applied to analysis of blood samples.
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Jagiellonian University, 3 Ingardena Street, 30-060 Kraków, Poland.
Aneta
A
Woźniakiewicz
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Jagiellonian University, 3 Ingardena Street, 30-060 Kraków, Poland. Electronic address: wietecha@chemia.uj.edu.pl.
Renata
R
Wietecha-Posłuszny
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Jagiellonian University, 3 Ingardena Street, 30-060 Kraków, Poland.
Michał
M
Woźniakiewicz
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Jagiellonian University, 3 Ingardena Street, 30-060 Kraków, Poland.
Julia
J
Nowak
Laboratory for Forensic Chemistry, Department of Analytical Chemistry, Jagiellonian University, 3 Ingardena Street, 30-060 Kraków, Poland.
Paweł
P
Kościelniak
D001569:Benzodiazepines / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D008872:Microwaves; D015203:Reproducibility of Results; D014116:Toxicology / Q000379:methods
D001569:Benzodiazepines
Benzodiazepines; Human serum and blood analysis; Liquid chromatography; Mass spectrometry; Microwave-assisted extraction