目的: 建立汉麻超高效液相色谱（UHPLC）指纹图谱及大麻二酚酸（CBDA）、大麻二酚（CBD）、四氢大麻酚（THC）、四氢大麻酚酸（THCA）4个成分含量同时测定的分析方法,为汉麻的质量评价提供依据。方法: 采用Phenomenex C18 色谱柱（150 mm×4.6 mm,2.6 μm）,流动相为乙腈-0.2%磷酸水溶液,流速为1.0 mL · min-1,柱温为35 ℃,检测波长为220 nm,进样量为2 μL,梯度洗脱;使用《中药色谱指纹图谱相似度评价系统（2012版）》对大兴安岭地区和齐齐哈尔地区的15批汉麻进行指纹图谱分析,采用聚类分析、主成分分析和正交偏最小二乘判别分析,对汉麻UHPLC指纹图谱进行分析评价。结果: 建立了汉麻UHPLC指纹图谱,标定了26个共有峰,指认了其中4个色谱峰并进行了含量测定,分别为CBDA、CBD、THC、THCA,其相似度均> 0.950。其中,大兴安岭地区样品聚为Ⅰ类,质量最好。表明不同产地汉麻样品有较大差异。26个共有峰经对照品比对,且由变量重要性投影可知,4个被指认成分均是导致汉麻差异的主要成分。结论: 该方法稳定可靠,可用于汉麻的定性定量分析,研究结果为汉麻的质量评价提供依据。
Objective: To establish the UHPLC fingerprint and quantitative analysis of CBDA, CBD, THC, and THCA for hemp, and provides a basis for the quantity control of hemp. Methods: UHPLC was launched on a Phenomenex C18 (150 mm×4.6 mm, 2.6 μm) with mobile phase of acetonitrile-0.2% phosphoric acid aqueous solution at a flow rate of 1.0 mL · min-1, column temperature of 35 ℃, detection wavelength of 220 nm, and an injection volume of 2 μL by gradient elution. 15 batches of hemp samples from Daxing ’anling region and Qiqihar were analyzed to establish the fingerprint. Cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were adopted to assess of UHPLC fingerprint of hemp. Results: the UHPLC fingerprint of hemp was established, and 26 common peaks were selected the 26 common peaks were compared with the reference substance. The structures of four chromatographic peaks were identified and simultaneously analyzed for content determination. four chromatographic peaks were identified as CBDA, CBD, THC and THCA. The similarity of fingerprint of them was greater than 0.950. The samples from Daxing’anling were clustered into class Ⅰ, with the best quality. The results show that hemp samples from different regions were quite different. The variable importance projection (VIP) showed that the four compounds to be tested were the main components leading to the difference of hemp. Conclusion: The established method is stable and reliable and can be used for qualitative and quantitative analysis of hemp. The findings of this study are expected to provide a scientific basis for quality control of hemp.
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