Research Progress on Biological Matrix Reference Materials in Forensic Toxicology.

Biological matrix reference material is a reference material that combines the target material with the biological matrix. The biological matrix reference material has higher consistency with the authentic specimens in forensic toxicology, and its application has a positive effect on improving the accuracy of test results. This paper reviews the research on the matrix reference materials corresponding to three common biological test materials (blood, urine and hair). In order to provide reference for the development and application of biological matrix reference materials in forensic toxicology, this paper mainly introduces the research progress of preparation technology of biological matrix reference materials and some existing products and their parameters evaluation.

Delayed elimination in humans after ingestion of colchicine: Two fatal cases of colchicine poisoning.

Colchicine has been widely used in the treatment of acute gout over the years, but it has a narrow therapeutic index, and overdose can be life threatening. A method using ultra-high-performance liquid chromatography-tandem mass spectrometry system was applied in two fatal cases of colchicine poisoning in this study to the determination of colchicine in blood. In case 1, a 19-year-old man suffered from depression and ingested 160 colchicine tablets (each 0.5 mg). The concentration of colchicine in his blood samples showed a fluctuating trend and kept above the therapeutic steady-state concentration for 5 days. In case 2, a 70-year-old female patient with a history of gout and chronic colchicine intake ingested five times the usual dose of colchicine (5 mg) and died after 12 days of medical care, with 5 ng/mL of colchicine in her blood sample. Our findings suggest that the delayed elimination and accumulation in humans after colchicine overdose could keep the concentration of colchicine maintaining above the therapeutic steady-state concentration for many days before dying, probably along with a fluctuating trend.

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.

Detection of amfepramone and its metabolite cathinone in human hair: Application to a uthentic cases of amfepramone use.

In recent years, overseas anti-obesity drugs including amfepramone have flowed into China through the internet or personal import by travelers. Amfepramone is controlled in China and is not available as a pharmaceutical product. It is obtainable either through the internet or imported by individuals across the border. The abuse of amfepramone is causing serious health problems. A method for the detection and quantification of amfepramone and its metabolite cathinone in human hair was developed and fully validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Approximately 10 mg of hair was weighed and pulverized with extraction solvent (a mixture of methanol: acetonitrile: 2 mM ammonium formate [pH 5.3] [25:29:46, v/v/v]). The limit of detection (LOD) and the limit of quantitation (LOQ) were 5 and 10 pg/mg, respectively. The method was linear over a concentration range from 10 to 10,000 pg/mg. The accuracy varied from -9.3% to 2.3%, with acceptable intra- and inter-day precision. The validated method was successfully applied to 17 authentic cases. The amfepramone concentrations ranged from 11.7 to 209 pg/mg, with a median of 30.2 pg/mg, and the hair cathinone concentrations ranged from 11.9 to 507 pg/mg, with a median of 54.0 pg/mg. This is the first report of amfepramone concentrations in human hair from amfepramone users. Cathinone can be incorporated into hair after amfepramone use.

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.

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.

Simultaneous Quantitative Determination of Amphetamines, Opiates, Ketamine, Cocaine and Metabolites in Human Hair: Application to Forensic Cases of Drug Abuse.

A method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to simultaneously quantify amphetamines, opiates, ketamine, cocaine, and metabolites in human hair is described. Hair samples (50 mg) were extracted with methanol utilizing cryogenic grinding. Calibration curves for all the analytes were established in the concentration range 0.05-10 ng/mg. The recoveries were above 72%, except for AMP at the limit of quantification (LOQ), which was 48%. The accuracies were within ±20% at the LOQ (0.05 ng/mg) and between -11% and 13.3% at 0.3 and 9.5 ng/mg, respectively. The intraday and interday precisions were within 19.6% and 19.8%, respectively. A proficiency test was applied to the validated method with z-scores within ±2, demonstrating the accuracy of the method for the determination of drugs of abuse in the hair of individuals suspected of abusing drugs. The hair concentration ranges, means, and medians are summarized for abused drugs in 158 authentic cases.