基于超高效液相色谱-质谱联用技术对大麻植物中3种成分及化学表型分析

Determination of Three Kinds of Cannabinoids in Cannabis Using Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry and Analysis of Phenotype of Cannabis

建立了超高效液相色谱-质谱联用(UPLC/PDA-QDa)同时对大麻植物中Δ9-四氢大麻酚(Δ9-THC)、大麻酚(CBN)和大麻二酚(CBD)进行定性与定量分析的方法。缴获的大麻植物用甲醇超声萃取,采用甲醇(含0.1%甲酸)和超纯水为流动相,等度洗脱,流速为0.2 m L/min,经Waters UPLC BEH C_(18)柱(50 mm×2.1 mm,1.7μm)分离,利用光电二极管阵列检测器(PDA)在220 nm波长下检测,并通过质谱检测器(QDa)对目标洗脱峰进行追踪确证。在0.5~20μg/m L浓度范围内,3种大麻酚类化合物的质量浓度与峰面积呈良好的线性关系,R≥0.999;低、中、高添加水平的平均回收率为82%~102%,相对标准偏差(RSD)在0.4%~4.1%之间。本方法稳定、简便、灵敏,能够满足检测需求。根据Δ9-THC、(Δ9-THC+CBN)/CBD、Δ9-THC/CBD或CBN/CBD表型指数,区分不同产地大麻的化学表型,为大麻植物的检测分析和质量控制提供了有效手段。

An analytical method of ultra-high performance liquid chromatography-tandem mass spectrometry( UPLC/PDA-QDa) for the qualitative and quantitative analysis of Δ9-tetrahydrocannabinol( Δ9-THC),cannabidiol( CBD),and cannabinol( CBN) in confiscated cannabis was developed. The seized cannabis was extracted in methanol by sonication. The binary mobile phase consisted of methanol( containing 0.1% formic acid) and water. After centrifugation,the supernatant was separated on Waters UPLC BEH C_(18) column( 50mm×2. 1 mm,1. 7 μm) with isocratic elution at a flow rate of 0. 2 m L/min. The three cannabinoids were analyzed by photodiode array( PDA) detector at 220 nm and confirmed by mass spectrometer QDa. The correlation coefficient of standard curve for the three cannabinoids in linearity range was not less than 0.999,as well as the recoveries were 82%-102% with the relative standard deviations( RSD) of 0.36%-4.12% at three spiked levels. The method is specific,easy,quick and suitable for confirmation of the cannabinoids in seized cannabis. Cannabis plants in different areas were classified by their chemical phenotype as drug-type or fiber-type plants, taking into account the phenotypic index Δ9-THC,(Δ9-THC+CBN)/CBD, or the Δ9-THC/CBD and the(Δ9-THC+CBN)/CBD ratios. The analysis of the original composition of plant material is necessary for the detection and the quality control of cannabis plants.