Simplified Analysis of 11-Hydroxy-Delta-9-Tetrahydrocannabinol and 11-Carboxy-Delta-9-Tetrahydrocannabinol in Human Meconium: Method Development and Validation

Abstract

We describe the development of a sensitive analytical method for the analysis of 11-hydroxy-delta-9-tetrahydrocannabinol (11-OH-THC) and 11-carboxy-delta-9-tetrahydrocannabinol (THCC) in meconium using a gas chromatography-mass spectrometry (GC/MS) platform. The method was validated according to protocols, which included assessment of accuracy, precision, robustness, stability in meconium and in-process stability, interference and sensitivity and specificity. The method consists of a solid phase extraction with alkaline hydrolysis and derivatization of the analytes with N, O-Bis(trimethylsilyl)trifluoroacteamide, followed by GC/MS analysis using selected ion monitoring. The method uses deuterated internal standards for both analytes. Calibration curves had r(2) values >0.998, and extraction efficiency was determined to be 84.7% for THCC and 78.6% for 11-OH-THC. The detection limit for both analytes was 5 ng/g. This confirmatory method was successfully applied to 183 meconium samples that had screened positive by enzyme-linked immunosorbent assay, and 67.2% were confirmed for THCC, and 2.2% were confirmed positive for 11-OH-THC. The mean (SD) and median (range) THCC (n = 123) concentrations detected were 55.0 ng/g (±59.0) and 33.75 ng/g (5-265 ng/g), while the mean and median (range) for 11-OH-THC (n = 4) concentrations were 8.25 ng/g (±4.71) and 6.5 ng/g (5-15 ng/g).

Screening Pregnant Women and Their Neonates for Illicit Drug Use: Consideration of the Integrated Technical, Medical, Ethical, Legal, and Social Issues

“…Another LC-MS/MS method has been published capable of screening 22 antidepressant and anxiolytic drugs (Pichini et al, 2016), as well as a method for screening 20 antiretroviral drugs and metabolites in a single analytical run (Himes et al, 2013). The majority of available methods for meconium screening are LC-MS based, and many others exist with shorter run times, screening drugs and metabolites specific to certain drug classes such as marijuana, nicotine, or alcohol use (Tynon et al, 2015; Prego-Meleiro et al, 2017). …”

Monitoring Perinatal Exposure to Cannabis and Synthetic Cannabinoids

“…Meconium testing potentially provides information on drug exposure over the last trimester of pregnancy and the first few days after delivery. For this reason, the detection of cannabinoids in meconium became the most common method for documenting in utero cannabis exposure 39,41,[44][45][46]48,49,51 . Immunoassay screenings and gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmatory methods were used for meconium cannabinoid analyses.…”

“…Immunoassay screenings and gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmatory methods were used for meconium cannabinoid analyses. Given the volatile and lipophilic nature of cannabi-noids, GC-MS has been the gold standard for quantification, using phenyl-, methyl-, and dimethylpolysiloxane columns 39,44,46,48,49,51 . However, the emergence of sensitive LC-MS/MS technologies requiring less time-consuming sample preparations (no derivatization) is now common (C18 and pentafluorophenylpropyl columns) 41,45 .…”

“…Prego-Meleiro et al 41 and Kim et al 45 quantified THCCOOH-glucuronide in meconium samples and further confirmed the extensive glucuronidation of THCCOOH. The most recent studies involving meconium cannabinoid detection used hydrolysis under basic conditions to improve THCCOOH detection capability 39,44 . CBD and CBN were also proposed as meconium prenatal cannabis exposure markers, although with generally lower concentrations than THCCOOH 41,45,51 .…”

Avaliação toxicológica da exposição à >i<Cannabis>/i< e cocaína na gravidez em cordão umbilical humano: validação de método analítico e prospecção de biomarcadores proteicos de toxicidade

“…Os processos de triagem são realizados com ensaios imunoenzimáticos, e podem apresentar como principal problema uma baixa especificidade e sensibilidade analítica para drogas comuns. Outro viés é a ausência de dispositivos de triagem para uma ampla gama de drogas, levando, em suma, à aplicação de métodos de confirmação (AGIUS; NADULSKI, 2014).…”