Research And Application Of Cannabis And Its Major Metabolite Integrated Detection Method

Objective: To Establish an integrated detection techniques and methods of marijuana and its metabolites in human biological samples(urine,blood).Method: 1.preliminary screening by the colloidal gold for suspected biological samples.2.human samples(urine,blood)after hydrolysis solution is made weakly acidic aqueous solution containing a deuterated THC(THC-D3)and containing deuterated THCA(THC-COOH-D3)as an internal standard reagent using solid phase extraction(SPE)method to extract biological sample tetrahydrocannabinol(THC),THC acid(THC-COOH),cannabinol(CBN),cannabidiol(CBD),the silylating agent after BSTFA(containing 1% TMCS)derivatized,GC-MS determination in the relative retention times(RRT).3.confirm biological samples qualitative by Contrast standards to relative retention times(RRT)and Mass spectrum;analytes and the internal standard peak area ratio as ordinate series,standard solution concentration as abscissa standard curve for quantitation analysis.Results: Blood samples: Tetrahydrocannabinol(THC),Cannabinol(CBN),Cannabidiol(CBD),THCA(THC-COOH)had good linear relationship with the peak area within 10 ~ 80 ng/mL,correlation coefficient(R2)> 0.996 7,the average recovery rate is 91.8% ~ 97%,the relative standard deviation(RSD)is 0.59% ~1.8%.content of THC in 5 cases of blood sample were 12,19,13,30,23 ng/mL.content of CBN in 3 cases of blood were 43,47,56 ng/m L,content of CBN in 2 cases of blood is < 40 ng/m L.content of CBD in 2 cases of blood were 55,60 ng/mL,content of CBD in 3 cases of blood is < 50 ng/m L.content of THC-COOH in 4 cases of blood sample is 47,50,61,86 ng/m L,content of THC-COOH in 1 cases of blood sample is < 25ng/mL.Urine samples: Tetrahydrocannabinol(THC),Cannabinol(CBN),Cannabidiol(CBD),THCA(THC-COOH)had good linear relationship with the peak area within 10 ~ 400 ng/m L,correlation coefficient(R2)> 0.997 4,the average recovery rate is 93.1% ~ 99.3% and the relative standard deviation(RSD)is 0.27% ~ 1.16%,As most of the THC acidification formed THC-COOH,there is no content of THC or less than the limit of detection in 5 cases of urine sample.content of CBN in 1 cases of urine is 41 ng/mL,content of CBN in 1 cases of urine is < 40 ng/m L,content of CBN in 3 cases of urine is < 20 ng/mL.content of CBD in 3 cases of urine were 71,56,50 ng/mL,content of CBD in 1 cases of urine is < 50 ng/m L,content of CBD in 1 cases of urine is < 15 ng/m L.content of THC-COOH in 5 cases of urine is 88.4,99,53,47,65 ng/mL.Conclusion: The method is simple,accurate,simultaneous detection and quantification of tetrahydrocannabinol(THC),THC acid(THC-COOH),cannabinol(CBN),cannabidiol(CBD)and other active ingredients of marijuana.