一测多评法测定花椒及相关产品中4个酰胺类物质的含量Simultaneous determination of four amides in Zanthoxylum bungeanum Maxim. and its related products with quantitative analysis of multi-components by single marker

• Published in: Zhongguo you zhi Vol. 50; no. 9; pp. 131 - 138
• Main Author: 张雪梅1,2，舒晓1,2，张伟1,2，谢蔓莉1,2，杨晓霞1,2，杨丰庆3，杨俊英1,2 ZHANG Xuemei1,2, SHU Xiao1,2, ZHANG Wei1,2, XIE Manli1,2, YANG Xiaoxia1,2,YANG Fengqing3, YANG Junying1,2
• Format: Journal Article
• Language: English
• Published: 中粮工科（西安）国际工程有限公司 01.09.2025
• Subjects: zanthoxylum bungeanum maxim. and its related products; zanthoxylum amides; high-performance liquid chromatography; quantitative analysis of multi-components by single marker; relative correction factor; 花椒及花椒制品；花椒酰胺；高效液相色谱；一测多评法；相对校正因子
• ISSN: 1003-7969
• DOI: 10.19902/j.cnki.zgyz.1003-7969.240581

为了对花椒及相关产品中酰胺类化合物准确定量，以羟基-α-山椒素为内参物，建立了一测多评法同时测定花椒及相关产品中羟基-α-山椒素、羟基-β-山椒素、羟基-γ-山椒素和羟基-ε-山椒素4个成分含量的方法。采用高效液相色谱法（外标法）建立了4个成分的标准曲线，并对外标法进行了方法学考察。在外标法的基础上，以羟基-α-山椒素为内参物，获得其他3个成分的相对校正因子和相对保留时间，以相对保留时间对样品峰进行定性，以相对校正因子和外标法获得的样品中羟基-α-山椒素含量计算花椒及相关产品中其他3个成分的含量。对一测多评法的适用性进行了考察，并以35个实际样品（干花椒、鲜花椒、花椒粉、花椒油、花椒叶）为例，对比了外标法和一测多评法测定结果的差异。结果表明：外标法4个成分的标准曲线回归方程线性关系良好，具有较好的精密度、稳定性、重复性和准确性；在各自线性范围内，羟基-α-山椒素与羟基-β-山椒素、羟基-γ-山椒素和羟基-ε-山椒素之间的相对校正因子分别为0.539、0.245和0.541；高效液相色谱仪和色谱柱对羟基-β-山椒素、羟基-γ-山椒素和羟基-ε-山椒素的相对校正因子和相对保留时间影响很小，柱温及流动相组成对相对校正因子的影响也很小；采用一测多评法和外标法测得实际样品中4个成分的含量值接近，相对误差绝对值在0~5.56%之间。综上，所建立的一测多评法简便、可靠、重现性好，适用于花椒及相关产品中4个酰胺类物质含量的同时测定。 In order to accurately quantify amides in Zanthoxylum bungeanum Maxim. and its related products, the method of quantitative analysis of multi-components by single marker (QAMS) was established using hydroxy-α-sanshool as the internal reference substance for the simultaneous determination of hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-γ-sanshool, and hydroxy-ε-sanshool. The standard curves of the above four components were established by high-performance liquid chromatography (external standard method), and the methodology of the external standard method was investigated. Based on the external standard method, the relative correction factors and relative retention time of the other three components were obtained using hydroxy-α-sanshool as the internal reference substance. The relative retention time was used for qualitative analysis of the sample peaks, and the contents of the other three components in Zanthoxylum bungeanum Maxim. and its related products were calculated based on the relative correction factors and the content of hydroxy-α-sanshool in the samples obtained by the external standard method. The applicability of the QAMS method was investigated. Taking 35 actual samples (dried Zanthoxylum bungeanum, fresh Zanthoxylum bungeanum, Zanthoxylum bungeanum powder, Zanthoxylum bungeanum oil, and Zanthoxylum bungeanum leaves) as examples, the differences between the determination results of the external standard method and the QAMS method were compared. The results showed that the standard curves of the four components established by the external standard method had good linear relationships and had better precision, stability, repeatability, and accuracy. Within their respective linear ranges, the relative correction factors between hydroxy-α-sanshool and hydroxy-β-sanshool, hydroxy-γ-sanshool, and hydroxy-ε-sanshool were 0.539, 0.245 and 0.541, respectively. The influences of the high-performance liquid chromatography instrument and chromatographic column on the relative correction factors and relative retention time of hydroxy-β-sanshool, hydroxy-γ-sanshool, and hydroxy-ε-sanshool were very small, and the influences of column temperature and mobile phase composition on the relative correction factors were also very small. The contents of the four components in the actual samples measured by the QAMS method and the external standard method were close, with the absolute value of relative error ranging from 0 to 5.56%. In conclusion, the established QAMS method is simple, reliable, and reproducible, and is suitable for the simultaneous determination of the contents of the four amides in Zanthoxylum bungeanum Maxim. and its related products.