UHPLC-MS/MS method for simultaneously detecting 16 tryptamines and their metabolites in human hair and applications to real forensics cases.
Yan Shi, Roujia Wang, Shuai Yuan, Huosheng Qiang, Min Shen, Baohua Shen, Olaf H Drummer, Zhiguo Yu, Yunli Zhao, Ping Xiang
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences November 30, 2020 DOI: 10.1016/j.jchromb.2020.122392 via PubMed
Summary
A rapid UHPLC-MS/MS method was developed to screen 16 tryptamines in hair samples. Twenty milligrams of hair were pulverized and processed, achieving limits of detection from 0.1 to 20 pg/mg and quantitation from 3 to 50 pg/mg. The method was applied to 191 hair samples from suspected tryptamine users; 77 tested positive for 5-MeO-DiPT, while the other 114 showed none of the targeted tryptamines or metabolites. In 34 samples, 5-MeO-DiPT and its metabolites 5-MeO-NiPT, 5-OH-DiPT, and 4-OH-DiPT were detected together. The method demonstrated good accuracy (91-114%), precision, and recovery (85-115%), making it suitable for forensic analysis of tryptamine use.
Study at a glance
| Characteristics | Method development and validation Peer reviewed |
|---|---|
| Sample size | 191 |
| Population | Suspected tryptamine users providing hair samples |
| Keywords | 5-MEO-Dipt Hair analysis Metabolites Tryptamine Uhplc-ms/ms |
| Key finding | The validated UHPLC-MS/MS method successfully detected 5-MeO-DiPT in 77 of 191 hair samples, with its metabolites also identified in 34 samples. |
Abstract
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.