| Driving under the influence of drugs (DUID) has become the widespread concerned issue because of its huge dangers. Psychotropic drugs due to the ADR (Adverse Drug Reaction) will affect driving. Determination of the concentration of psychotropic drugs in human blood will helpful for public transport safety. Considering a large number of samples to be determined, the assay should meet not only the requirements of the analysis methodology, but also high detection efficiency, timeliness and other issues.At present, China Public Security Industry Standards only have GA/T1073-2013 ?HS-GC assay for simultaneous determination of ethanol, methanol, n-propanol, acetaldehyde, acetone, isopropanol, n-butanol in human blood or urine?, but without testing standards for DUID. The current assay for determination of biological samples can not be applied in DUID, because there are some flaws. The aim of the present study was to establish a unified, fast method for DUID. Here, we established a LC-MS/MS assay for simultaneous determination of 25 psychotropic drugs (Alprazolam, Amitriptyline, Carbamazepine, Chlorpromazine, Chlorpheniramine, Clonazepam, Clozapine, Diazepam, Doxepin, Estazolam, Lozazepam, Ketamine, Methamphetamine, Midazolam, Morphine, Nitrazpeam, Oxazepam, Perphenazine, Phenacetin, Promethazine, Triazolam, Trifluoperazine, Zaleplon, Zolpidem, Zopiclone) in human blood, and applied the method to analyze 3140 blood samples from drivers suspected traffic offense in Hangzhou from Junel,2014 to December 31,2014. Additionally, we also explored the effect of ethanol on the concentrations of ketamine in blood of mice after oral concomitant administration of ethanol and ketamine.1.LC-MS/MS assay for simultaneous determination of 25 psychotropic drugs in human bloodObjectives To establish a LC-MS/MS assay for simultaneous determination of 25 psychotropic drugs in human blood. Methods Acetonitrile was added to the blood sample for precipitating the protein, the mixture was centrifuged, the supernatant was dried, and the residue was reconstituted. After centrifugation, the supernatant was analyzed with LC-MS/MS. The separation was performed on C18 chromatography column (50mm*3.0mm*2.6?m),the mobile phase was consisted of 0.1% formic acid (phase A), acetonitrile:methanol=1:1 (phase B) with gradient elution. The results revealed the calibration curves for 25 psychotropic drugs were linear over the ranges of 0.05?20.0 ng/mL with R=0.9944?0.9996, the extract recoveries were 83.0%? 99.7%,the assay recoveries were 80.2%?97.4%,the intraday and interday precious were 1.6%?14% and 3.1%?14%.Conclusion The method is sensitive, repeatable and accurate for simultaneous determination of 25 psychotropic drugs in human blood.2.Concentrations of 25 psychoactive drugs in whole blood samples from drivers suspected traffic offense in HangzhouObjectives To depict the situation of motor vehicle drivers and psychotropic substances abuse. Methods 3140 blood samples from drivers suspected traffic offense in Hangzhou during June 1,2014 to December 31,2014 were determined by the assay developed, the relationship of the determination results and source information of the samples, including gender, age, and other information were analyzed. Results 116(3.7%) samples were found containing at least one drug, in which 91 samples contained chlorpheniramine within the range of 6.6?19.2 ng/mL. Conclusion Chlorpheniramine is the most positive detected drug, which probably is attributing to the easy acquiring because of its OTC sell, however its ADR is not fully understood by drivers.3.The effect of ethanol on the blood concentration of ketamine in miceObjectives Study the effect of ethanol on blood concentration of ketamine in miceafter oralconcomitant administration of ethanol and ketamine. Methods Male ICR mice were orally administered ketamine at low(10mg/kg), medium(30mg/kg), high(50mg/kg), alone or concomitant administration of low(320mg/kg), medium (640mg/kg), high(960mg/kg) dose of ethanol, respectively. Blood samples were collected at 30min, 1h,2h after dose. The concentration of ketamine was determined by LC-MS/MS. Results 1) The concentration of ketamine in mouse blood increased as the dosage raising; 2) Compared with the ketamine alone giving, concomitant administration of ethanol significantly raised the concentration of ketamine. However, the dosage of ethanol at low, medium and high did not show the different effect.Conclusion Alcohol can increase the concentration of ketamine in mouse but without dose-dependent within 320?960mg/kg. |
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