OBJECTIVES: Tiletamine, a veterinary anesthetic, has emerged as a novel psychoactive substance and has been abused in many parts of the world, causing great harm to public health. However, the sensitivity of existing detection methods cannot meet the needs of forensic practice. This study aims to establish an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of tiletamine and its metabolite desethyltiletamine in human biological samples, and to verify its applicability in forensic practice. METHODS: SKF_525A was used as the internal standard. Biological samples were extracted with acetonitrile containing 1 ng/mL SKF_525A, vortexed for 10 min, ultrasonicated for 20 min, centrifuged at 10 000 r/min for 10 min, and 500 μL of the supernatant was filtered through a 0.22 μm membrane. Analyses were performed using an ACQUITY UPLC H-Class PLUS system and an XEVO TQ-S Micro triple quadrupole mass spectrometer. An ACQUITY UPLC^® BEH C18 (1.7 µm, 2.1 mm×100 mm) column at a flow rate of 0.3 mL/min was used, and four mobile phase systems were tested to optimize separation. Detection used positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode, with quantifier ion transitions of mass to charge 224.043→179.016 for tiletamine and mass to charge 196.08→151.06 for desethyltiletamine. Calibration curves were established over 0.1-200 ng/mL in spiked blood samples. The linear range, limit of detection (LOD), and limit of quantification (LOQ) were determined. Low (5 ng/mL), medium (20 ng/mL), and high (100 ng/mL) concentrations of tiletamine were spiked into blood, liver, and kidney to evaluate precision, accuracy, matrix effect, recovery, and stability. Finally, actual forensic case samples were tested to validate applicability. RESULTS: The established UPLC-MS/MS method achieved simultaneous detection of tiletamine and desethyltiletamine in human biological samples, with retention times of 3.42 min and 2.82 min, respectively. Using mobile phase A (20 mmol/L ammonium acetate and 0.1% formic acid in water) and mobile phase B (acetonitrile) produced the best separation. In blood, tiletamine showed good linearity from 0.1-200 ng/mL (r=0.992, R²=0.983), LOD 0.03 ng/mL, LOQ 0.1 ng/mL, recovery 92%-107%, and matrix effect 71%-99%. In liver and kidney, recoveries were 91%-98% and 93%-104%, and matrix effects were 69%-96% and 72%-100%, respectively. Intra- and inter-day precision [expressed as relative standard deviation (RSD)] and accuracy [expressed as relative error (RE)] were within 15%, and samples were stable at -20 ℃. Tiletamine was detected in actual case samples at 0.37 μg/mL (blood), 0.15 μg/g (liver), 0.11 μg/g (kidney) in case 1, and 8.75 ng/mL (blood) in case 2; desethyltiletamine was also detected in blood. CONCLUSIONS The UPLC-MS/MS method is efficient, accurate, and sensitive, and is suitable for detecting tiletamine and desethyltiletamine in human biological samples.
Tandem Mass Spectrometry/methods* ; Humans ; Chromatography, High Pressure Liquid/methods* ; Substance Abuse Detection/methods* ; Liquid Chromatography-Mass Spectrometry

Objective To establish accurate and rapid methods to identify four new synthetic cannabinoids （JWH-203, JWH-122, 5F-APINACA and AB-CHMINACA） in blood samples. Methods The whole blood samples were extracted by acetonitrile and methanol, screened by gas chromatography-mass spectrometry （GC-MS） then confirmed by liquid chromatography-tandem mass spectrometry （LC-MS/MS） method, and multiple reaction monitoring （MRM） mode was used for quantitative analysis. Results The GC-MS method needed 21 min to complete the analysis, while the LC-MS/MS method needed 5 min. The AB-CHMINACA, JWH-203, 5F-APINACA and JWH-122 all used quasi molecular ion peak as a parent ion. The precursor-product ion combinations were m/z 357.4→312.2, m/z 340.2→125.0, m/z 384.1→135.1 and m/z 356.4→169.2. The four synthetic cannabinoids in blood samples had good linearity in the 1-250 ng/mL mass concentration range （r>0.99）. The limits of detection （LODs） were in the range of 0.1-0.5 ng/mL, the recovery rate was 85.4%-95.2%, the RSD less than 10.0%, and the matrix effect was 80.3%-92.8%. Conclusion The GC-MS and LC-MS/MS chromatographic behaviors and mass spectrometry analysis information of four synthetic cannabinoids were obtained in this study, and the possible causes of differences in chromatographic behaviors were discussed preliminarily. Therefore this study has a suggestive effect on judging the development trend of synthetic cannabinoids. This method can be used for rapid identification of four synthetic cannabinoids in blood, which can provide reference for identification of new synthetic cannabinoids when they are proliferating at present.
Blood Chemical Analysis/methods* ; Cannabinoids/blood* ; Chromatography, Liquid ; Humans ; Limit of Detection ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry

OBJECTIVES: To identify the new designer drugs which are totally unknown and not in the routine testing list by the technologies such as high-resolution mass spectrometry in drug facilitated sexual assault, in order to solve the problem in actual cases. METHODS: The milky fluid from an actual case was extracted and analyzed using LC-QE, ¹H-NMR and GC-MS, respectively. The accurate masses and cluster ions isotope patterns of unknown compound were obtained by LC-QE. The molecular formula was confirmed as C₁₆H₁₂C₂N₂O based on the protons number of ¹H-NMR. The isomers diclazepam and 4-chlorodiazepam were separated and detected with GC-MS. RESULTS: The new designer benzodiazepine as diclazepam in the milky fluid was identified. The results provided direct evidence for the investigation and qualitative analysis of such cases. CONCLUSIONS The combined application of various methods, including LC-QE, ¹H-NMR and GC-MS, can be used to detect unknown new psychoactive substances.
Benzodiazepines/chemistry* ; Benzodiazepinones ; Chromatography, Liquid/methods* ; Designer Drugs/chemistry* ; Female ; Gas Chromatography-Mass Spectrometry/methods* ; Humans ; Male ; Mass Spectrometry/methods* ; Sex Offenses ; Substance Abuse Detection/methods* ; Toxicology/methods*

OBJECTIVES: To infer the frequency of dosage and medication history investigate of the victims in drug facilitated cases by the segmental analysis of clonazepam in hair. METHODS: Freezing milling under liquid nitrogen environment combined with ultrasonic bath was used as sample pretreatment in this study, and liquid chromatography-tandem mass spectrometry was used for the segmental analysis of the hair samples collected from 6 victims in different cases. The concentrations of clonazepam and 7-aminoclonazepam were detected in each hair section. RESULTS: Clonazepam and its metabolite 7-aminoclonazepam were detected in parts of hair sections from the 6 victims. The occurrence time of drug peak concentration was consistent with the intake timing provided by victims. CONCLUSIONS Segmental analysis of hair can provide the information of frequency of dosage and intake timing, which shows an unique evidential value in drug facilitated crimes.
Adult ; Chromatography, Liquid ; Clonazepam/analysis* ; Crime ; Forensic Medicine/methods* ; Forensic Toxicology ; Hair/chemistry* ; Humans ; Mass Spectrometry ; Spectrometry, Mass, Electrospray Ionization ; Substance Abuse Detection/methods* ; Ultrasonics

OBJECTIVE: To establish a screening and confirmation method for psychotropic drugs and their metabolites in human blood and urine by HPLC-LTQ Orbitrap MS. METHODS: The samples were pretreated with Sirocco protein precipitation plate, and then analyzed by HPLC-LTQ Orbitrap MS. The method was validated in terms of the limit of detection (LOD). An accurate mass database was created for psychotropic drugs screening. RESULTS: The LOD for most of 56 determined compounds was < or = 0.1 ng/mL. The accurate mass database included the accurate mass information of 61 psychotropic drugs. CONCLUSION The method is accurate, rapid, sensitive and the database is suitable for psychotropic drugs screening and confirmation.
Chromatography, High Pressure Liquid/methods* ; Forensic Toxicology ; Humans ; Mass Spectrometry/methods* ; Molecular Structure ; Molecular Weight ; Psychotropic Drugs/urine* ; Reproducibility of Results ; Sensitivity and Specificity ; Substance Abuse Detection/methods*

OBJECTIVE: To compare morphine and codeine concentration in urines of heroin abusers and codeine users and to discuss the judgment index to distinguish between heroin abuser and codeine user. METHODS: The urines of heroin abusers and codeine users were collected at different time periods. After protein precipitation, the urine samples were conducted for the qualitative and quantitative analysis of morphine and codeine by UPLC-MS/MS. And the results were all statistically analyzed. RESULTS: Statistical analysis showed that morphine and codeine concentration in urines of heroin abusers and codeine users were both abnormal distributions. The probability of the heroin abuser would be more than 95% and less than 5% for the codeine user when the concentration of morphine in urine sample was more than 67 ng/mL. The probability of the codeine user would be more than 95% and less than 5% for the heroin abuser when the concentration of morphine in urine sample was less than 67 ng/mL. CONCLUSION The morphine concentration in urine could be used as a criterion to distinguish the heroin abuser from the codeine user, while the codeine concentration could not.
Administration, Oral ; Chromatography, High Pressure Liquid ; Codeine/urine* ; Forensic Toxicology/methods* ; Heroin Dependence/urine* ; Humans ; Morphine/urine* ; Solutions ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry/methods*

The paper reports the establishment of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneous analysis of 11 opiates in hair samples, and the study of presence of opiates in the hair of active heroin addicts. About 20 mg of decontaminated and pulverized hair sample was hydrolyzed with buffer solution for 30 min, in the presence of morphine-d3 and acetylmorphine-d6 used as internal standards, and then extracted with the mixture of dichlormethane and isopropanol, separated by the Allure PFP propyl column with a mobile phase consisting of acetonitrile and 20 mmol L(-1) ammonium acetate buffer, and then analyzed by LC-MS/MS. Multiple reaction monitoring (MRM) mode was used to analyze 11 opiates. Eleven opiates showed a fairly good linearity over the corresponding range (r > 0.996 0). The detection limits were less than 0.05 ng mg(-1). The recoveries were between 47.2% and 110%, and the deviations of intra- and inter-day precision were less than 14%. Heroin, acetylmorphine, morphine, codeine, acetylcodeine and hydrocodone were detected in hair samples of 21 herion addicts. The developed method shows high sensitivity and selectivity, and is suitable for the simultaneous analysis of 11 opiates in hair samples and identify legal and illegal use of opiates.
Analgesics, Opioid ; analysis ; Chromatography, Liquid ; methods ; Codeine ; analogs & derivatives ; analysis ; Hair ; chemistry ; Heroin ; analysis ; Humans ; Hydrocodone ; analysis ; Limit of Detection ; Morphine ; analysis ; Morphine Derivatives ; analysis ; Sensitivity and Specificity ; Substance Abuse Detection ; methods ; Tandem Mass Spectrometry ; methods

Due to the diversity of toxicologically relevant substances, the uncertainty of target compounds and the specificity of samples, toxicological screening techniques have always been valued by the forensic toxicologists. Depending on its powerful separation ability, superhigh resolution and accurate mass measurement, combined with the two levels spectrum database matching and abundance ratio of isotope ion, the liquid chromatography-high resolution mass spectrometry (LC-HRMS) analyzers have increasingly advantage in screening and identification of chemical compound. This review focuses on the applications of LC-HRMS in screening and identification of drug-of-abuse, prescription drugs, pesticide and stimulant. The prospect of LC-HRMS in forensic toxicology analysis is also included.
Central Nervous System Agents/analysis* ; Chromatography, Liquid/methods* ; Doping in Sports ; Forensic Toxicology/methods* ; Humans ; Pesticide Residues/analysis* ; Pesticides/analysis* ; Sensitivity and Specificity ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry/methods* ; Toxicity Tests/methods*

OBJECTIVE: To propose a method for simultaneous determination of codeine(COD), 6-monoacetyl-morphine (6-MAM), morphine (MOR), morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in human urine by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). METHODS: After precipitation of protein by acetonitrile, the urine samples, with added the morphine-d3 (MOR-d3) and morphine-3-Glucuronide-d3 (M3G-d3) as internal standards, were pre-treated by Sirocco protein precipitation plate, and then analyzed by UPLC-MS/MS. RESULTS: The limit of detection was 0.2 ng/mL for both COD and MAM, the limit of quantitation was 0.5 ng/mL for both COD and MAM. The limit of detection was 0.5 ng/mL for MOR, M3G and M6G, the limit of quantitation was 1 ng/mL for them. The linear correlation coefficients were not less than 0.9997, both the inter-day and intra-day precisions were less than 10%, the recoveries were in the range of 70.0% to 98.3%, the matrix effects were about 50.5% to 99.0%. CONCLUSION This proposed method is simple, rapid and accurate, it could be applied in forensic toxicological analysis.
Chromatography, Liquid/methods* ; Codeine/urine* ; Humans ; Limit of Detection ; Morphine/urine* ; Morphine Derivatives/urine* ; Reproducibility of Results ; Sensitivity and Specificity ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry/methods*

OBJECTIVE: To develop a method for determining ethyl glucuronide (EtG) in blood and urine by liquid chromatograph coupled with tandem mass spectrometer (LC-MS/MS). METHODS: After blood and urine de-proteined by acetonitrile, the supernate obtained from a centrifuge was analyzed by LC-MS/MS. RESULTS: Determination limit of EtG in both blood and urine was 0.05 pg/mL, with a linear range of 0.10-5.00 microg/mL (r > 0.999). Accuracy in both matrixes was 95%-109%. Inter- and intra-day RSD were less than 12%. The method showed an excellent performance when it was used to analyze authentic blood and urine samples for EtG. CONCLUSION The method is capable for blood and urine EtG analysis.
Alcoholism/urine* ; Biomarkers/urine* ; Chromatography, Liquid/methods* ; Ethanol/metabolism* ; Forensic Toxicology/methods* ; Glucuronates/urine* ; Humans ; Reproducibility of Results ; Sensitivity and Specificity ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry/methods*