Direct injection ultra-performance liquid chromatography-tandem mass spectrometry for the high-throughput determination of 11 illicit drugs and metabolites in wastewater.

The measurement of illicit drugs in wastewater is increasingly being adopted as a method for objective monitoring of population-level illicit drug use. This work describes the first small-volume direct-injection ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 11 illicit drugs and metabolites in wastewater. The method required an injection volume of only 30 µL of wastewater sample, the limits of detection (LOD) ranged from 0.4 ng/L to 2 ng/L and the lower limits of quantitation (LLOQ) ranged from 1 ng/L to 5 ng/L. Application of the method to real wastewater samples collected from wastewater treatment plants revealed morphine in all samples, together with other illicit drugs (methamphetamine, codeine, ketamine, and nor-ketamine) in some samples. Small-volume direct injection showed great potential as an efficient method for the high-throughput determination of illicit drugs in wastewater.

Forensic Analysis of 9 Poisoning Death Cases Caused by Oral Administration of Diphenidol.

OBJECTIVES: To analyze the characteristics of diphenidol poisoning cases and to provide clues and technical means for the identification of such cases. METHODS: Biological samples of 9 deaths caused by diphenidol poisoning were detected by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and the characteristics of these cases were analyzed retrospectively. RESULTS: Most of the deaths caused by diphenidol poisoning were young females. The dosage was between 60 and 300 tablets, and the mass concentration of diphenidol in the postmortem blood ranged from 0.87 to 99.00 µg/mL. There was no correlation between the dosage and the concentration of diphenidol in the blood. CONCLUSIONS: Diphenidol poisoning has the characteristics of high concealment and lethality. More attention should be paid to suicide cases, and diphenidol should be recommended as a routine detection item to avoid missing detection.

Strategies for Collection and Analysis of Samples in Simple Asphyxiant Gas Acute Poisoning Death Cases.

At present, the death cases of simple asphyxiant gas acute poisoning are increasing sharply. Common asphyxiant gases in death cases include nitrogen, helium, carbon dioxide, methane, propane, laughing gas, etc. Simple asphyxiant gas has no affinity for biological matrices and escapes quickly, which puts forward new requirements for autopsy procedures, selection and collection of samples, laboratory analysis and identification. This paper reviews the research and development process of death cases caused by simple asphyxiant gas acute poisoning and put forwards the collection and analysis strategy of the samples in such cases. The most valuable biological samples in such cases should be lung tissues associated with the airways, followed by brain tissue and cardiac blood. Gaseous samples from the esophageal cavity, tracheal cavity, pulmonary bronchi, gastric and cardiac areas are also recommended as valuable samples. In the case of postmortem examination, the gas should be injected into gas sample bag directly. Biological materials such as tissue and blood should be directly sealed in head-space vials and analyzed by using the headspace gas chromatography-mass spectrometry.

Analysis of 28 hair samples from users of the hallucinogenic beverage ayahuasca.

Ayahuasca is a psychoactive beverage widely used in religious ceremonies in Amazonia. Dimethyltryptamine is the main active compound of ayahuasca. Dimethyltryptamine has many hazardous effects, including hallucinations. In the present study, a fast and reliable UPLC-MS/MS method was developed and validated for the quantitation of dimethyltryptamine in hair samples. Twenty-milligram hair samples were pulverized with methanol below 4 °C. After ultrasonication, centrifugation and filtration, 200 µL of supernatant was placed into an autosampler vial for LC-MS/MS analysis. The lower limit of quantitation (LLOQ) was 3 pg/mg. The resulting calibration curve for dimethyltryptamine fit the expression y = 281.50213x + 0.00231 (R2 = 0.992). Acceptable intraday and interday precision (RSD < 15%) and accuracy (92-113%) were achieved. The dilution integrity was deemed acceptable based on accuracy (96%) and precision (1.8%). The validated method was successfully applied to 28 forensic cases. The concentrations of dimethyltryptamine ranged from 3 to 1109 pg/mg.

Application of a Validated UPLC-MS-MS Method for the Determination of Diphenidol in Biological Samples in 15 Authentic Lethal Cases.

Diphenidol (DPN) is a nonphenothiazinic antivertigo and antiemetic drug that has been widely used in clinical practice in China because of its good antivertigo curative effect, minimal side effects and high safety. In recent years, there have been some cases of sporadic suicide and accidental poisoning related to DPN. Hence, a validated method for the determination of DPN in biological samples by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) was developed. The method is characterized by the use of a simple, fast and inexpensive liquid-liquid extraction (LLE) for sample preparation, a rapid run time (5 min) and a low required sample volume (0.1 mL or 0.1 g). The lower limits of quantitation (LLOQs) were 0.05 ng/mL and 0.3 ng/g for blood and liver tissue, respectively. The method was shown to be linear over a concentration range of 0.05-200 ng/mL (blood) and 0.3-400 ng/g (liver). The accuracy was in the range of 92.77-112.75%. The relative standard deviations of the intraday and interday imprecisions were in the range of 3.22% to 12.17%, and the recoveries were in the range of 58.75-95.27%. Furthermore, the method was successfully applied to the detection and quantification of DPN in 15 real forensic cases. The postmortem concentration range of heart blood was 0.87-99 µg/mL.

Detection of a New Tert-Leucinate Synthetic Cannabinoid 5F-MDMB-PICA and Its Metabolites in Human Hair: Application to Authentic Cases.

Methyl 2 -[ [ 1- (5- fluoropentyl) indole - 3- carbonyl] amino] -3, 3- dimethyl - butanoate (5F-MDMB-PICA) is a new synthetic cannabinoid characterized by valinate or tert-leucinate moieties. In recent years, 5F-MDMB-PICA has been abused in the form of "spice-like" herbal incenses or electronic cigarette oil. A UHPLC-MS/MS method was developed to detect 5F-MDMB-PICA and its metabolites in human hair. Approximately 20 mg of hair was weighed and pulverized with methanol below 4°C. After ultrasonication, centrifugation and filtration, 200 µL of supernatant was placed into an autosampler vial and analyzed on a Waters Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 µm particle size) using an acetonitrile-20 mmol/L ammonium acetate (0.1% formic acid, 5% acetonitrile) gradient with a run time of 8 min. The limit of detection (LOD) ranged from 0.5 to 5 pg/mg, and the lower limit of quantitation (LLOQ) ranged from 1 to 5 pg/mg. The method was shown to be linear over a concentration range of 1-200 pg/mg. The linear correlation (R 2) of the calibration curves for all analytes was >0.999. The accuracy varied from 95.4 to 107.4%, while the intra- and inter-day precision RSD values were 0.7-10.6% and 1.7-12.2%, respectively. Recoveries were within the range of 61.1-93.3%, and matrix effects were in the range of 19.1-102.6%. The validated method was successfully applied to the identification and quantification of 5F-MDMB-PICA and its metabolites in hair from authentic forensic cases.

UHPLC-MS/MS method for simultaneously detecting 16 tryptamines and their metabolites in human hair and applications to real forensics cases.

Tryptamines are hallucinogenic substances many of which have appeared recently as novel psychoactive substances (NPS). Herein, we describe the establishment of a rapid UHPLC-MS/MS quantitative method for the targeted screening of 16 tryptamines of abuse in hair. Twenty milligram pieces of hair were pulverized below 4 °C in 0.5 mL of deionized water containing 0.1% formic acid and an internal standard (2 ng/mL psilocin-d10 and psilocybin-d4). After subsequent centrifugation, 5 µL of the supernatant was injected into a LC-MS/MS system fitted with a Waters Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 µm). The column was gradient eluted at 0.3 mL/min with mobile phases composed of 20 mmol/L ammonium acetate, 5% acetonitrile, and 0.1% formic acid in water (solvent A) and acetonitrile (solvent B). Limits of detection ranged between 0.1 and 20 pg/mg, with limits of quantitation ranging from 3 to 50 pg/mg. The calibration curves for all analytes were linear (r > 0.992). Accuracies varied between 91% and 114%, with intraday precision RSDs < 14% and interday precision RSDs of between 1.3% and 14%. The recoveries of all tryptamines were in the 85-115% range, with the matrix effect ranging from 95% to 112%. The validated method was successfully used to analyse 191 hair samples from suspected tryptamine users, 77 of which were 5-MeO-DiPT-positive, while the 16 tryptamines and their metabolites were not detected in the remaining 114 hair samples. 5-MeO-DiPT and its 5-MeO-NiPT, 5-OH-DiPT, and 4-OH-DiPT metabolites were concurrently detected in 34 hair samples. 5-MeO-DiPT, as the parent drug, was the parent substance found in the hair samples.

Determination of 37 fentanyl analogues and novel synthetic opioids in hair by UHPLC-MS/MS and its application to authentic cases.

The recent emergence of new fentanyl analogues and synthetic opioids on the drug market poses a global public health threat. However, these compounds cannot typically be identified using existing analytical methods. In this study, we aimed to develop and validate a rapid and sensitive method based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of 37 fentanyl analogues and novel synthetic opioids in hair samples. Hair samples (20 mg) were extracted by cryogenic grinding in an extraction medium of methanol, acetonitrile, and 2 mmol/L ammonium acetate (pH 5.3). Following centrifugation of the samples, the analytes were separated using a WATERS Acquity UPLC HSS T3 column. The limits of detection (LODs) and limits of quantification (LOQs) ranged from 0.5 to 2.5 pg/mg and from 2 to 5 pg/mg, respectively. The intraday and interday precisions were within 13.32% at LOQ, low, medium, and high levels. The accuracies were within the range of 85.63-116.1%. The extraction recoveries were in the range of 89.42-119.68%, and the matrix effects were within the range of 44.81-119.77%. Furthermore, the method was successfully applied to the detection and quantification of fentanyl and sufentanil in hair samples from two authentic cases. Thus, this method has great potential for detecting fentanyl analogues and novel synthetic opioids in forensic work.

Segmental analysis of antidepressant and antipsychotic drugs in the hair of schizophrenic patients.

Hair analysis is useful for documenting long-term exposure to drugs. The potential of hair analysis for therapeutic drug monitoring within the forensic field has been studied, but reference values for some antidepressants and antipsychotics in the hair of individuals undergoing chronic therapy are still lacking. In the present study, a method was developed and validated for the determination of 23 analytes, including antidepressants, antipsychotics, and related metabolites, in human hair by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Hair samples (10 mg) were extracted with a 25:25:50 (v/v/v) mixture of methanol/acetonitrile/2 mM ammonium formate (8% acetonitrile, pH 5.3) utilizing cryogenic grinding. The present method demonstrated sufficient selectivity, robustness, and accuracy. Sixteen analytes in hair were reported in 46 psychiatric patients receiving fixed drug dosages. To the best of our knowledge, the hair concentrations of perphenazine and norolanzapine, as well as the concentrations of amisulpride, aripiprazole and its metabolite dehydroaripiprazole, olanzapine, and sulpiride, in hair from individuals receiving fixed dosages is reported for the first time. A significant relationship between the administered dose and the concentration in the proximal hair segment was found only for clozapine, norclozapine, and chlorpromazine. The results confirmed that the idea of using hair concentrations to monitor a daily dose is inapplicable.

Screening for volatile sulphur compounds in a fatal accident case.

Acute fatal poisoning due to the inhalation of toxic gas frequently occurs in China. Volatile sulphur compounds (VSCs) are toxic to humans. In fatal poisoning investigations, such as those in industrial settings, a number of VSCs, including methanethiol (MT), dimethyl sulphide (DMS), dimethyl disulphide (DMDS) and dimethyl trisulphide (DMTS), can coexist. To date, there is limited data regarding these compounds in post-mortem cases. In the present study, we report toxicological findings in a fatal accident case with two victims. Headspace gas chromatography/flame ionization detector with two columns of different polarities was utilized to screen MT, DMS, DMDS and DMTS in blood. The limits of detection in both methods were 0.05 mg/mL. No sulphur compounds were detected in the blood samples of the two victims. DMS and DMDS were detected in the lungs at concentrations of 0.5 and 1.3 mg/g and 2.2 and 4.1 mg/g, respectively. DMDS liver concentrations were 2.5 and 6.5 mg/g. In addition to hydrogen sulphide, screening for additional VSCs could help establish the cause of death.