基于HPLC检测花椒和火锅底料中5种山椒素

Determination of 5 Sanshools in Zanthoxylum Bungeanum and Hotpot Seasoning by High-performance Liquid Chromatography

目的 基于高效液相色谱(HPLC)建立检测花椒和火锅底料中5种山椒素含量的方法,对比提取溶剂、料液比、超声时间和超声次数对花椒和火锅底料中山椒素的提取效果差异。方法 花椒和火锅底料分别以甲醇溶液提取,经C18-WP(5μm,4.6 mm×250 mm)色谱柱分离,以甲醇和水溶液为流动相进行梯度洗脱,流速1.0 m L/min,柱温40℃,检测波长270 nm,进样量20μL。结果 花椒中山椒素的提取条件为样品与甲醇按料液比1∶15(g/m L)混合,超声处理30 min,重复提取3次;火锅底料中山椒素的提取条件为,样品与甲醇按料液比1∶4(g/m L)混合,超声处理30 min,重复提取2次。在上述色谱条件下,5种山椒素在0.4~100μg/m L范围内线性关系良好,相关系数r≥0.999 3,检出限为0.001 0~0.040 0 mg/g;在3个不同浓度添加水平下,花椒中各目标物质的加标回收率在76.99%~85.28%范围内,火锅底料中各目标物质的加标回收率在72.55%~86.88%范围内,两者的相对标准偏差(RSD)均<10%。结论 花椒与火锅底料的前处理条件存在差异,这与两者的样品差异有关,花椒与火锅底料的组成不同、山椒素含量不同,所以山椒素的提取条件会存在一定差异。该检测方法具有良好的准确性和精密度,可用于测定花椒和火锅底料中5种山椒素的含量。

Objective This study aimed to develop a method for determining five sanshools in Zanthoxylum bungeanum and hotpot seasonings through high-performance liquid chromatography(HPLC).The differences in extraction effects of sanshools in Zanthoxylum bungeanum and hotpot seasoning caused by different extraction solvents,solid-liquid ratios,ultrasonic time,and ultrasonic frequencies were compared.Methods In this study,samples were extracted with methanol and then purified on a C18-WP column(5μm,4.6 mm×250 mm).Water and methanol solutions were used as the elution phase,and gradient elution was performed at a flow rate of 1.0 mL/min,column temperature of 40℃,detection wavelength of 270 nm,and injection volume of 20μL.Results The extraction conditions for sanshools in Zanthoxylum bungeanum were as follows:the sample was mixed with methanol at the ratio of 1∶15(g/mL),ultrasonic treatment for 30 minutes,and repeated extraction 3 times;the extraction conditions for sanshools in hotpot seasoning were as follows:the sample was mixed with methanol at the ratio of 1∶4(g/mL),ultrasonic treatment for 30 minutes,and repeated extraction 2 times.Under the above chromatographic conditions,the linear relationship of the five sanshools was good in the range of 0.4~100μg/mL,with a correlation coefficient of r≥0.9993 and detection limits of 0.0010~0.0400 mg/g.At three different concentration levels,the spiked recovery rate of the target substances in Zanthoxylum bungeanum was in the range of 76.99%~85.28%,and in hotpot seasoning was in the range of 72.55%~86.88%,and the relative standard deviation(RSD)of both was less than 10%.Conclusion There are differences in the pre-treatment conditions of Zanthoxylum bungeanum and hotpot seasoning,which are related to the differences in the samples.Due to the different compositions and sanshool contents in Zanthoxylum bungeanum and hotpot seasoning,the extraction conditions of sanshools will vary.This detection method has good accuracy and precision and can be used to determine the content of five sanshools in Zanthoxylum bungeanum and hotpot seasoning.