Fast confirmation for marijuana metabolite: THC-COOH, ultra-fast LC—MS/MS run time, and application to routine samples

Abstract

Summary.A fast, reliable, inexpensive, and practical method with a low determination limit and high recovery has been developed for the determination of the marijuana metabolite in routine analysis. THC-COOH in urine was validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Before an easy single-step extraction with Toxi-Tubes, basic hydrolysis was performed at 60 °C for 30 min. LC-MS/MS analysis takes 2.5 min for each sample, and the retention time of the analyte is 1.75 min. Specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, repeatability, and intermediate precision (inter-day) system suitability parameters were determined in the validation study. The recovery of the extraction method was 88.67 (±5.91). LOD and LOQ values were 1.41 and 5.00 ng mL −1 , respectively. The method showed linear response between the values 5.00 and 500.00 ng mL −1 . The repeatability was 9.64% (relative standard deviation, RSD%), and the intermediate precisions (RSD R %) were 10.73%, 13.74%, and 8.11% at 10.00, 100.00, and 200.00 ng mL −1 concentration levels, respectively. No statistically significant difference was found in ANOVA analysis, between three consecutive days in intermediate precision study, for 90% confidence level. HorRat values were between 0.34 and 0.61. The method was applied to CEDIA positive samples, obtained from the Trabzon Group Presidency of Turkish Council of Forensic Medicine, successfully.

A review on the recent advances in HPLC, UHPLC and UPLC analyses of naturally occurring cannabinoids (2010–2019)

“…Marijuana or hashish, a cocktail of at least 30 different major cannabinoids, generally prepared from crushing the leaves, flowers (inflorescenses) and even stems of C. sativa , is one of the oldest recreational and addictive natural products used by humans for centuries . However, the non‐medical use of Cannabis or marijuana is illegal in many countries, which prompts the use of analytical tools, like HPLC and UPLC, to analyse biological and forensic samples like blood, oral liquid, hair and urine, to confirm marijuana usage (Tables ) . After consumption of marijuana, THCA ( 13 ), which is present in the crude marijuana, but converted to THC ( 12 ) by heat during smoking, is excreted in the urine as its glucuronide conjugate, and can be analysed comfortably by LC methods.…”

“…Human urine samples are popular in forensic toxicological analysis for the determination of various illegal drugs, including cannabinoids, and their metabolites . Table summarises the applications of HPLC and UPLC‐based analytical methods for naturally occurring cannabinoids and their biotransformation products in human urine samples , . Detection techniques used in these studies included simple UV, PDA and MS/MS methods.…”

“…A rather much shorter column (20 mm × 2.1 mm; particle size: 2.5 μm) reversed‐phase silica C 18 analytical column achieved successful separation and ESI‐MS/MS based quantitative detection of CBD ( 3 ), CBN ( 8 ), THC‐COOH ( 10 ) and THC ( 12 ) . There have been several other ESI‐MS/MS based HPLC analytical methods reported for the analysis of cannabinoids and their biotransformation products in human urine samples (Table ) . While the length of most of the HPLC columns used in these studies was between 50 mm and 250 mm, a much shorter SunFire C 18 column (20 mm) was used by Lendoiro et al ., where a simple isocratic elution with 0.1% aqueous HCOOH and ACN, with a total run time of 5 min, was applied.…”