黑龙江汉麻叶中化学成分研究与大麻二酚（CBD）含量测定

Study on chemical constituents and determination of cannabidiol (CBD) in hemp leaves of Heilongjiang province

摘要:

研究黑龙江汉麻叶中相关化学成分，建立大麻二酚（CBD）含量检测方法.采用硅胶柱色谱、C18反相中压柱及半制备色谱对汉麻叶乙酸乙酯提取物进行分离纯化，得到5个单体化合物，经核磁NMR解析确定结构为：大麻二酚（CBD）、大麻萜酚（CBG）、大麻酚（CBN）、Δ9-四氢大麻酚酸（Δ9-THCA）、大麻色烯（CBC）；以石油醚-二氯甲烷（1∶1）为展开剂，采用碘和1%香草醛乙醇硫酸（10∶1）分别显色，建立了CBD薄层色谱（TLC）检测方法；以乙腈-0.05%磷酸溶液（80∶20）为流动相，选择CBD吸收峰较高的检测波长，建立了CBD高效液相色谱（HPLC）检测方法；该方法在3.125~100 μg/mL线性关系良好（R2=0.9997）,平均回收率为103.69%，RSD 1.32%（n=5）.

Abstract:

The purpose of this research is to study the chemical constituents in the leaves of Heilongjiang han hemp and establish a method for the determination of cannabidiol (CBD). The ethyl acetate extract of hemp leaf was separated and purified by silica gel column chromatography, C18 reverse phase medium voltage column and semi preparative chromatography to obtain 5 monomer compounds. And the structure of these compounds was determined by nuclear magnetic (NMR) analysis. They were cannabidiol (CBD), cannabigerol (CBG), cannabinol (CBN), Δ9-Tetrahydrocannabinolic acid (Δ9-THCA) and cannabichromene (CBC). By using petroleum ether dichloromethane (1∶1) as the developing solvent, iodine and 1% Vanillin ethanol sulfuric acid (10∶1)were used to establish the TLC method for CBD respectively. High performance liquid chromatography (HPLC) detection method for CBD was also established by using acetonitrile 0.05% phosphoric acid solution (80∶20) as mobile phase and selecting the detection wavelength with high CBD absorption peak, which has a good linear relationship at 3.125~100μg/ mL (R2=0.9997), the average recovery rate is 103.69% and RSD is 1.32% (n=5).