黑龙江汉麻叶中化学成分研究与大麻二酚(CBD)含量测定
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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3.125～100μg/mL线性关系良好(R~2=0.9997),平均回收率为103.69%,RSD 1.32%(n=5).
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 3.125～100 μg/mL(R~2=0.9997), the average recovery rate is 103.69% and RSD is 1.32%(n=5).
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 3.125～100 μg/mL(R~2=0.9997), the average recovery rate is 103.69% and RSD is 1.32%(n=5).
引文
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[16] Hayakawa K,Mishima K,Abe K,et al.Cannabidiol prevents infarction via the non-CB1 cannabinoid receptor mechanism [J].Neuroreport,2004,15:2381.
[2] 王建新.化妆品天然成分原料介绍(Ⅵ)[J].日用化学品科学,2018(12):52.
[3] Ligresti A,Morielloa S,Starowicz K,et al.Antitumor activity of plant cannabinoids with emphasis on the effect of cannabidiol on human breast carcinoma[J].J Pharmacol Exp Ther,2006,318:1375.
[4] 马约内,罗西,盖伊,等.用于治疗神经性疼痛的大麻素类化合物:CN201280024106.4 [P].2014-01-22.
[5] 郭蓉,陈璇,郭鸿彦.四氢大麻酚和大麻二酚的药理研究进展[J].天然产物研究与开发,2017(08):189.
[6] 卢延旭,董鹏,崔晓光,等.工业大麻与毒品大麻的区别及其可利用价值[J].中国药理学通报,2007,23:1112.
[7] 翟晚枫,张春水,高利生.高效液相色谱外标工作曲线法测定大麻树脂中四氢大麻酚含量的不确定度评定[J].化学分析计量,2012,21:8.
[8] 高宝昌,孙宇峰,张旭,等.工业大麻叶中大麻二酚含量分析研究[J].黑龙江科学,2018(1):61.
[9] 艾斯凯尔·艾尔肯,孙力扬,谢辉,等.气相色谱-质谱联用(GC-MS)法检测人体血液、尿液中大麻及其主要代谢物的含量[J].新疆医科大学学报,2016,39:1020.
[10] Choi Y H,Hazekamp A,Peltenburg-Looman A M G,et al.NMR assignments of the major cannabinoids and cannabiflavonoids isolated from flowers of Cannabis sativa [J].Phytochem Anal,2004,15:345.
[11] Kemal M,Khalil S K W,Rao N G S,et al.Isolation and identification of a cannabinoid-like compound from Amorpha species [J].J Nat Prod,1979,42:463.
[12] Marjolein S,Georgios A,Jve E,et al.Development of a cannabinoid-based photoaffinity probe to determine the δ8/9-tetrahydrocannabinol protein interaction landscape in neuroblastoma cells [J].2018,3:136.
[13] Wang G,Zhu L,Zhao Y,et al.A natural product from Cannabis sativa subsp.sativa inhibits homeodomain-interacting protein kinase 2 (HIPK2),attenuating MPP(+)-induced apoptosis in human neuroblastoma SH-SY5Y cells[J].Bioorg Chem,2017,72:64.
[14] Teare L,Zajicek J.The use of cannabinoids in multiple sclerosis [J].Expert Opin Investig Drugs,2005,14:859.
[15] Hayakawa K,Mishima K,Abe K,et al.Cannabidiol prevents infarction via the non-CB1 cannabinoid receptor mechanism [J].Neuroreport,2004,15:2381.
[16] Hayakawa K,Mishima K,Abe K,et al.Cannabidiol prevents infarction via the non-CB1 cannabinoid receptor mechanism [J].Neuroreport,2004,15:2381.
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