Development of a rapid column-switching LC-MS/MS method for the quantification of THCCOOH and THCCOOH-glucuronide in whole blood for assessing cannabis consumption frequency

Abstract

The concentration of 11-nor-9-carboxy-Δ 9 -tetrahydrocannabinol (THCCOOH) in whole blood is used as a parameter for assessing the consumption behavior of cannabis consumers. The blood level of THCCOOH-glucuronide might provide additional information about the frequency of cannabis use. To verify this assumption a column-switching liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and direct quantification of free and glucuronidated THCCOOH in human whole blood was newly developed. The method comprised protein precipitation, followed by injection of the processed sample onto a trapping column and subsequent gradient elution to an analytical column for separation and detection. The total LC run time was 4.5 min. Detection of the analytes was accomplished by electrospray ionization in positive ion mode and selected reaction monitoring using a triple stage quadrupole mass spectrometer. The method was fully validated by evaluating the following parameters: linearity, lower limit of quantification, accuracy and imprecision, selectivity, extraction efficiency, matrix effect, carry-over, dilution integrity, analyte stability and re-injection reproducibility. All acceptance criteria were analyzed and the predefined criteria met. Linearity ranged from 5.0 -500 µg/L for both analytes. The method was successfully applied to whole blood samples from a large collective of cannabis consumers, demonstrating its applicability in the forensic field.

Assessing cannabis consumption frequency: Is the combined use of free and glucuronidated THCCOOH blood levels of diagnostic utility?

“…Based on these findings, we hypothesize that THCCOOH‐glucuronide blood concentration is a valuable supplemental marker for assessing the frequency of cannabis consumption and that the combined use of THCCOOH and THCCOOH‐glucuronide blood levels, while retaining maximum specificity, will increase the diagnostic sensitivity for the detection of regular cannabis users. To test this hypothesis, we recently developed a liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) method for the rapid and simultaneous quantification of THCCOOH and THCCOOH‐glucuronide in blood …”

“…All drivers considered for this evaluation had detectable THCCOOH levels (≥2.5 µg/L), which identified them as cannabis consumers. To determine THCCOOH‐glucuronide and total THCCOOH concentrations, the blood samples were re‐analyzed with our recently developed LC‐MS/MS method …”

“…Whole blood THCCOOH concentrations were measured within a few days after blood collection by GC‐ or LC‐MS/MS as described by Fabritius et al . For the present study, the occasional and heavy smokers' blood samples were thawed after storage at ‐80 °C and were re‐analyzed by a recently developed LC‐MS/MS method for rapid and simultaneous quantification of free and glucuronidated THCCOOH . Briefly, 100 μL of whole blood samples were aliquoted into 96‐well plates and proteins were precipitated by adding 300 μL of cold acetonitrile containing the two deuterated internal standards.…”

Accelerated quantification of amphetamine enantiomers in human urine using chiral liquid chromatography and on-line column-switching coupled with tandem mass spectrometry

“…Method validation was conducted according to international guidelines [20,21] and as previously reported [22]. Linearity was assessed by analyzing duplicate calibration curves on three different days (n = 6).…”

Desenvolvimento de fase monolítica com grupos amino e nano partículas para a determinação de carbidopa, levodopa, benserazida, dopamina e 3-O-metildopa em amostras de plasma de pacientes com a doença de Parkinson por HILIC-MS/MS no modo column switching

“…Sistema column switching em modo foreflush e backflush. Adaptado de(MARCHIONI et al, 2017).Diferentes fases seletivas têm sido utilizadas como coluna 1D no sistema column switching, dentre elas podemos destacar as fases monolíticas (DOMINGUES; SOUZA; QUEIROZ, 2015; MIZUNO; KATAOKA, 2015; LUO; LI; HU, 2017), polímeros molecularmente impressos -MIP -(XU et al, 2010a), materiais de acesso restrito -RAM -(ECKERT;GÖEN, 2014;FAGUNDES et al, 2014;ZHOU et al, 2014;ZHANG et al, 2015; CAGLAR ANDAC, 2016), colunas de fluxo turbulento (CRUTCHFIELD; MARZINKE; CLARKE, HERVIOU et al, 2016;SHIN et al, 2016), imunosorventes (CHAVES; QUEIROZ, 2013), nanotubos de carbono(DE FARIA et al, 2017) e suportes de sílica -C18 -(PARK et al, 2014;PURSLEY et al, 2014;SABAREDZOVIC et al, 2015;XIONG et al, 2015;HÄDENER et al, 2016;ZHONG et al, 2016) A Tabela 2 ilustra algumas aplicações de diferentes fases seletivas na primeira dimensão do sistema LC no modo column switching. Diferentes fases seletivas na primeira dimensão do sistema LC no modo column switching para análise de diferentes analitos em amostras biológicas…”