Research On The Test Methods Of Two Synthetic Cannabinoids And Their Metabolites In Hair

Synthetic cannabinoids?SCs?which are high efficiency,long duration of action,and high addiction are a new class of synthetic compounds that are very similar to the physiological and pharmacological effects of Delta-9-Tetrahydrocannabinol??9-THC?,the main active ingredient in natural cannabis.At present,there are 251 synthetic cannabinoids?31.2%of known new psychoactive substances?,and 53 species have been regulated in China.In the"Decision on Amending the"Methods for Identifying Drug Addictions",which was implemented on April 1,2017,the detection of drug components in human hair samples as an important basis for public security organs to identify their drug addiction.On October 31,2018,the Ministry of Public Security of the People's Republic of China issued the"Specifications for the Detection of Hair Samples of Drug-Related Persons".Compared with conventional biological samples such as urine,blood,and tissue,hair samples have unique advantages.At present,there are no reports on synthetic cannabinoids and their major metabolites in hair.Therefore,there is an urgent need to establish an analytical method for the synthesis of cannabinoids and their major metabolites in hair.Based on the actual situation of public security,this paper selects two synthetic cannabinoids JWH-073?1-butyl-3-?1-naphthoyl?indole?and AM-2201?1-?5-fluoropentyl?-3-?1-naphthoyl?indole?that are common in the case,studies the metabolic mechanism and test methods of JWH-073 and AM-2201,and establishes high performance liquid chromatography-tandem mass spectrometry test methods which include JWH-073 and AM-2201 and its main metabolites JWH-073-OH and AM-2201-OH,so as to provide scientific basis and technical support for the identification of this type of drug addiction.The main research contents are as follows:The first chapter summarizes the various theories of drugs entering the hair,gives a comprehensive introduction to hair pretreatment technology,and lists its applications in related fields.Chapter 2 establishes an incubation model of liver microsomes in vitro,artificially simulates the metabolic environment in vivo,and analyzes the metabolites of synthetic cannabinoids by mass spectrometry information.The experimental results show that JWH-073and AM-2201 all produce 11 phase I metabolites in the vitro incubation model of liver microsomes.The main metabolic pathways are hydroxylation,carboxylation,dealkylation and carbonylation,and determine its main metabolites based on their response intensity.In chapter three,high-performance liquid chromatography-tandem mass spectrometry rapid detection methods for synthetic cannabinoids JWH-073 and AM-2201 and their main metabolites were established.A ZORBAX Ecilpse Pluse C₁₈?2.1 mm×100 mm,1.8?m?column was selected;0.1%formic acid-water?5 mmol/L ammonium acetate??phase A?and methanol?phase B?as mobile phases,flow rate 0.4 mL/min,injection volume 1?L,column temperature 35°C;using electrospray ion source Positive ion scan mode?ESI⁺?,multiple reaction monitoring mode?MRM?.Chapter 4 optimizes and compares three pretreatment methods for extraction of JWH-073and AM-2201 and their main metabolites in hair,acidic methanol sonication,alkaline digestion and acid digestion,and establish a method for the determination of JWH-073 and AM-2201 and their main metabolites JWH-073-OH and AM-2201-OH in hair by high performance liquid chromatography-tandem mass spectrometry.The two synthetic cannabinoids and their main metabolites have a good linear relationship in the range of 5⁵00 pg/mg?R²>0.996?;at low,medium and high addition levels,the recovery rate of acidic methanol ultrasonic extraction is86.4%-92.8%.The recovery of alkaline digestion was between 80.9%and 91.5%,and the recovery of acid digestion was between 62.3%and 78.8%.The detection limits?S/N=3?of JWH-073 and AM-2201 and their main metabolites are in the range of 0.1¹.0 pg/mg,and the limit of quantification?S/N=10?is in the range of 0.3⁴.0 pg/mg.Intra-day precision RSD?8.5%,and intra-day precision RSD?12.0%.Chapter 5 obtained a real hair sample through 5-week animal feeding.The real hair sample was used to verify the validity of the method and was applied to practical cases.