Determination Of Three Common Synthetic Cannabinoids In Body Fluids By Ultra-performance Liquid Chromatography-tandem Mass Spectrometry

Synthetic cannabinoids(SCs),which are a chemically diverse group of compounds functionally similar to THC,have attracted interest of many scientists in recent years because not only have more serve psychoactive effects,but also access availability.So they have been used around the world directly affects the health of human.The numbers of emergency cases caused by SCs have been increasing.They have been linked to a number of severe poisoning and death cases worldwide.At present,there are no reports on the test of SCs in biological samples.So it is necessary to establish a rapid and effective identification method.SCs are classified according to their time of occurrence,JWH-018 and JWH-250 belong to the first generation of SCs,AM-2201 belongs to the second generation of SCs,and they are all common and representative SCs.In this article established the method of sample preparation and detecting JWH-018,JWH-250 and AM-2201 in blood and saliva.The detailed steps as follows:1.Establish the analyzing method of JWH-018,JWH-250 and AM-2201.2.For the blood sample,three pretreatment methods including protein precipitation,QuEChERS,solid-phase extraction are investigated respectively.The advantages and disadvantages of those three pretreatment methods are compared and evaluated.Three pretreatment methods for the simultaneous determination of synthetic cannabinoids,including JWH-018,JWH-250 and AM-2201 in human blood samples by ultra-performance chromatography-tandem mass spectrometry(UPLC-MS/MS)have been established.3.For the saliva sample,three pretreatment methods including protein precipitation,QuEChERS,liquid-liquid extraction(LLE)are investigated respectively.The advantages and disadvantages of the three pretreatment methods are compared and evaluated.Three pretreatment methods for the simultaneous determination of synthetic cannabinoids,including JWH-018,JWH-250 and AM-2201 in human saliva samples by UPLC-MS/MS have been established.Results:1.Waters BEH C18 column by gradient elution with 0.1%formic acid-0.1%formic acid-acetonitrile as mobile phase,ionized with positive electrospray(ESI~+),detected under multiple reaction monitoring mode(MRM).The parent ion for JWH-018 was m/z 342.2.Two daughter ions were monitored at m/z 154.9 and 127.2.The parent ion for JWH-250 was m/z 336.0.Two daughter ions were monitored at m/z 121.0 and 91.0.The parent ion for AM-2201 was m/z360.2.Two daughter ions were monitored at m/z 155.0 and 127.1.The limits of detection(LODs)based on a S/N ratio of 3:1,JWH-018 was 0.05 ng/mL,JWH-250 and AM-2201 were0.01 ng/mL.The limits of quantitation(LOQ),based on a S/N ratio of 10:1,JWH-018 was 0.1ng/mL,JWH-250 and AM-2201 were 0.05 ng/mL.The calibration curve of JWH-018 was linear within the concentration range 0.1~50 ng/mL,the calibration curve of JWH-250 was linear within the concentration range 0.05~50 ng/mL and the calibration curve of AM-2201 was linear within the concentration range 0.05~40 ng/mL,the correlation coefficients(R~2)more than0.999 that indicates an excellent linearity.2.For the blood sample,the calibration graphs were linear in their concentration ranges with the correlation coefficients(R~2)more than 0.994.The average recoveries of protein precipitation method for target drugs were 62.0%to 81.3%.The average recoveries of QuEChERS for target drugs were 80.3%to 108.6%.The average recoveries of SPE for target drugs were 70.1%to 98.5%.The limits of detection(LOD)of three methods were in the range of 0.01~0.1 ng/mL(S/N=3),while limits of quantification(LOQ)were in the range of 0.05~0.2ng/mL(S/N=10).Inter-day precision and intra-day precision were less than 11.1%that indicates a good reproducibility.3.For the saliva sample,the calibration graphs were linear in their concentration ranges with the correlation coefficients(R~2)more than 0.998.The average recoveries of protein precipitation method for target drugs were 62.6%to 78.3%.The average recoveries of QuEChERS for target drugs were 80.0%to 89.5%.The average recoveries of LLE for target drugs were 87.7%to 97.9%.The limits of detection(LOD)of three methods were in the range of 0.01~0.08 ng/mL(S/N=3),while limits of quantification(LOQ)were in the range of0.05~0.15 ng/mL(S/N=10).Inter-day precision and intra-day precision were less than 9.3%that indicates a good reproducibility.