Application of a UPLC-MS/MS method for quantitative analysis of 29 synthetic cannabinoids and their metabolites, such as ADB-BUTINACA and MDMB-4en-PINACA in human hair in real cases.

In recent years, an increasing number of new synthetic cannabinoids have appeared on the drug trade market. Many of the new synthetic cannabinoids have not previously been reported. At present, there are relatively few methods available for detecting synthetic cannabinoids and their metabolites in hair matrices. Therefore, we established a simple and fast method to simultaneously identify 29 synthetic cannabinoids and their metabolites in human hair by UPLC-MS/MS. Twenty milligrams of hair was used and processed by cryo-grinding and extraction with methanol. A Waters Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 µm) was used for chromatographic separation. Mobile phase A was comprised of 20 mmol/L ammonium acetate, 0.1% formic acid, and 5% acetonitrile and water, and mobile phase B was acetonitrile. The method was fully validated and proved to have good selectivity, accuracy, precision, and satisfactory linearity within the calibrated range. The limit of detection (LOD) ranged from 0.5 to 5 pg/mg, and the lower limit of quantitation (LLOQ) ranged from 1 to 10 pg/mg. The extraction recovery was 36.1-93.3%, and the matrix effect was 19.1-110.0%. The validated method was successfully used to qualitatively and quantitatively analyze 29 synthetic cannabinoids and their metabolites in 59 actual hair samples. MDMB-4en-PINACA had the highest positive detection rate followed by ADB-BUTINACA, and there are multiple synthetic cannabinoid mixed ingestions. This methodology has great potential for the detection of 29 synthetic cannabinoids and their metabolites in forensic cases.

Investigation of Fragmentation Pathways of Fentanyl Analogues and Novel Synthetic Opioids by Electron Ionization High-Resolution Mass Spectrometry and Electrospray Ionization High-Resolution Tandem Mass Spectrometry.

The global drug market is characterized by the fast development of new psychoactive substances such as fentanyl analogues and novel synthetic opioids, the detection of which is complicated by the lack of appropriate quality control procedures and references. Herein, we analyze the fragmentation pathways and characteristic ions of 25 novel fentanyl analogues and 5 novel synthetic opioids by electron ionization (EI) and electrospray ionization (ESI) high-resolution mass spectrometry to provide a reference for the identification of these species. In the ESI mode, fentanyl analogues mainly undergo piperidine ring degradation, phenethyl and piperidine ring dissociation, and piperidine ring and amide moiety cleavage, while piperidine ring degradation and phenethyl and piperidine ring dissociation are the major pathways in the EI mode. The five novel synthetic opioids largely undergo amide group dissociation and N-cyclohexyl bond cleavage in the ESI mode. Thus, this work facilitates the detection and quantitation of fentanyl analogues and novel synthetic opioids or other substances with similar structures in forensic laboratories.

Evaluation of Brodifacoum-induced Toxicity by Metabonomics Approach Based on HPLC-TOF-MS / 法医学杂志

Objective T o analyse the m etabolic changes in urine of rats w ith brodifacoum intoxication, and to reveal the m olecular m echanism of brodifacoum-induced toxicity on rats. Methods B y establish-ing a brodifacoum poisoning rats m odel, the urine m etabolic profiling data of rats w ere acquired using high performance liquid chromatography-timeofflightmassspectrometry (HPLC-TOF-M S).The orthogo-nal partial least squares analysis-discrim ination analysis (O PLS-D A ) w as applied for the m ultivariate statistics and the discovery of differential m etabolites closely related to toxicity of brodifacoum . Results O PLS-D A score plot show ed that the urinary m etabolic at different tim e points before and after drug adm inistration had good sim ilarity w ithin tim e period and presented clustering phenom enon. C om paring the urine sam ples of rats before drug adm inistration w ith w hich after drug adm inistration, tw enty-tw o m etabolites related to brodifacoum-induced toxicity w ere selected. Conclusion T he toxic effect of brodi-facoum w orked by disturbing the m etabolic pathw ays in rats such as tricarboxylic cycle, glycolysis, sphin-golipid m etabolism and tryptophan m etabolism , and the toxicity of brodifacoum is characterized of accu-m ulation effect. The m etabonom ic m ethod based on urine H PLC-TO F-M S can provide a novel insight into the study on m olecular m echanism of brodifacoum-induced toxicity.

Determination of Sulfide Ion in Blood from Hydrogen Sulfide Poisoning Cases / 法医学杂志

Objective T o establish a gas chrom atography-m ass spectrom etry (G C-M S ) m ethod for the determ ination of sulfide ion in blood and apply it to the practical cases. Methods T he 1, 3, 5-tribro-m obenzene w as selected as an internal standard, and 0.2 m L blood sam ple w as collected and analyzed using G C-M S after α-B rom o-2, 3, 4, 5, 6-pentafluorobenzyl brom ide derivatization. Results T he m ass concentration of sulfide ion in blood had good linearity in the range of 0.2-40μg/m L w ith a lim it of detection (L O D ) of 0.05μg/m L . T he m ass concentration of sulfide ion w as less than 0.05μg/m L in blank blood from different sources such as healthy subjects and dead cases. In 3 sulfide poisoning cases, sul-fide ion w as detected in the blood sam ples of 6 victim s, and the m ass concentration range w as 1.02-3.13μg/m L . Conclusion T his study establishes a m ethod for investigation of sulfide ion in blood w hich has been applied successfully to the cases of fatal sulfide poisonings.

Chlorpyrifos Determined in Human Blood by UPLC-MS/MS and Its Application in Poisoning Cases / 法医学杂志

Objective To determine the chlorpyrifos in human blood by liquid chromatography-tandemmass spectrometry and to validate its application in poisoning cases. Methods The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 mG II column (250 mm×2.0 mm, 5μm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammoniumacetate) and solvent B (methanol with 2 mmol/L ammoniumacetate) at 5∶95 (V∶V).Results The linearranged from5 to 500ng/mL (r=0.9987).Thelimitofdetection (LOD) and the lower limit of quantification (LLOQ ) were 2 ng/mL and 4 ng/mL , respectively. For this method, the precision and accuracy of intra-day and inter-day were <10% and 97.44%-101.10%, respectively. The re-sults in stability test of long-termfrozen were satisfied. The matrix effect, recovery and process efficien-cy were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively. Conclusion This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.

Analysis of GHB and Its Precursors in Urine and Their Forensic Application / 法医学杂志

Objective To establish the m ethod to analyze γ-hydroxybutyric acid (GHB ) and its precursors 1,4-butanediol (1,4-B D ) and gam m a-butyrolactone (GBL) in urine through LC-M S/M S and provide evi-dence for related cases. Methods GHB-d6 and M O R-d3 were used as the internal standard. The urine sam ple w as separated by LC after protein precipitation w ith m ethanol. The electrospray ion source w as for ionization. E ach com pound w as detected through m ultiple-reaction m onitoring (M R M ) m ode. Results The lim its of detection of GHB and its precursors 1,4-B D and GBLwere 0.1, 0.1 and 2μg/m L. The accuracy w as 87.6% -98.1% . The intra-day and inter-day precisions were less than 15% and m atrix ef-fects were higher than 80% . Conclusion The m ethod is high sensitive, sim ple, rapid, specific and w ith high reliability. This study has provided technical support and basic data for forensic cases involving GHB .

Study on appraisement and determination GHB levels in hair / 法医学杂志

OBJECTIVE@#The possibility for the identification of GHB administration through hair analysis was investigated to provide method and information for toxicology examination of GHB. METHODS A GC/MS assay for GHB in hair was developed. Endogenous levels of GHB in hair, time course of GHB in hair, relationship between GHB levels in hair and hair color or administration dose were also established by guinea pig model.@*RESULTS@#Endogenous levels of GHB in guinea pig black hair and human black hair were (3.01 +/- 1.41) ng/mg (n=28) and (1.02 +/- 0.27) ng/mg (n=20), respectively. GHB levels in black hair were increased by GHB administration and related with drug dosage, and also much higher than in brown and white hair.@*CONCLUSION@#Analysis of GHB in hair is suitable for investigation of GHB abuse in forensic toxicology and GHB level in segmental analysis compared with endogenous level of GHB may provide useful information about GHB administration.

Effect of Ethanol on Pharmacokinetics of Phenytoin Sodium in Rabbits / 中国药房

OBJECTIVE:To study the effect of ethanol on the pharmacokinetics of phenytoin sodium in rabbits METHODS:The serum concentrations of phenytoin sodium at different points of time were determined by UV-spectrophotometry in eight rabbits after administration of phenytoin sodium(10mg/kg)alone and in combination with ethanol RESULTS:After administration in combination with ethanol,the AUC of phenytoin sodium was significantly decreased from(4 108 64?1 039 98)mg/(min L) to (1 903 65?1 003 40)mg/(min L),Cmax from(29 0?2 94)mg/L to(16 0?5 9)mg/L,T1/2(ke) from(98 45?26 4)min to(82 84?25 5)min;but the apparent distribution volume(Vd)was obviously increased from(0 3 475?0 0 360)L/kg to(0 6 819?0 1 901)L/kg and the clearance rate(CL) from(0 0 026?0 0 008)ml/(kg?min)to(0 0 062?0 0 022)ml/(kg?min) CONCLUSION:The elimination of phenytoin sodium was significantly accelerated after simultaneous administration of ethanol in rabbits