HPLC测定黑果枸杞鲜果中花青素含量的方法研究

束彤 于瑶 马明芳 陈睿

摘  要：目的：优化并建立黑果枸杞鲜果中HPLC的测定方法。方法：采用乙醇、盐酸、水为提取液，超声提取、水浴水解后用HPLC测定黑果枸杞鲜果中的花青素含量。对乙醇浓度、盐酸浓度、超声时间、水浴温度进行正交实验条件优化，对实验的检出限、线性关系、重复性及回收率进行了考察。结果：确定各因素影响主次顺序为水解温度>盐酸浓度>超声时间>乙醇浓度，最优提取条件为盐酸30%、乙醇50%、超声时间15min、沸水浴。本方法6种花青素的检出限都可达到12mg/kg，在1～50μg/mL范围内，除天竺葵色素R2=0.9989外，其余花青素R2均大于0.999，6次独立平行测定RSD均小于4%，6种花青素回收率在89.0%～91.7%。结论：该方法可以用于黑果枸杞鲜果中花青素含量的测定。

关键词：黑果枸杞鲜果;花青素;高效液相色谱;正交实验

中图分类号 TS264.4文献标识码 A文章编号 1007-7731（2019）01-0024-07

Determination of Anthocyanin in Fresh Fruit of Lycium ruthenicum Murr.by HPLC

Shu Tong et al.

（Qinghai Province Institute for Food Control，Xining 810000，China）

Abstract：Objective：A high performance liquid chromatographic （HPLC） method was established for the determination of anthocyanin in fresh fruit of Lycium ruthenicum Murr.，and the orthogonal method was use to research the optimum extraction conditions. Methods：Anthocyanin were extracted by ultrasonic （solvent extraction were ethanol，hydrochloric acid and water），extract was hydrolyzed into major anthocyanins in hot water bath，and then HPLC was used to test the content of anthocyanins. Four factors （concentration of ethanol，concentration of hydrochloric acid，ultrasonic time and water bath temperature） were optimized by orthogonal experiment. The limits of detection （LOD），linearity，repeatability and recovery of the method were studied. Result：The order of primary and secondary of extract factor were temperature of water bath，concentration of hydrochloric acid，ultrasonic time and concentration of ethanol，and optimal extraction conditions were 30% hydrochloric acid，50% ethanol，ultrasonic extract 15 min and boiling water bath;and the result showed that LOD could up to 12mg/kg of all 6 kinds of anthocyanins，the linearity of the method is good （R2 of pelargonidin was 0.9989，and the others'were all above 0.999，over the concentration range from 1 μg/mL to 50μg/mL ），the relative standard deviations （RSD） of 6 tests were all less than 4%，and the ranges of recoveries at different levels were 89.0%～91.7%. Conclusion：the method can be used to test anthocyanidins in fresh fruit of Lycium ruthenicum Murr..

Key words：Fresh fruit of Lycium ruthenicum Murr.;Anthocyanin;High performance liquid chromatographic （HPLC）;Orthogonal experiment

花青素，又名花色素，是一類水溶性的类黄酮化合物，广泛存在于植物的花、果实、茎、叶中[1]。花青素具有多种生理功能，包括抗氧化、抗辐射、抗衰老以及调节血脂功能，能有效防治动脉粥样硬化等慢性疾病[2-4]。本试验对黑果枸杞鲜果中花青素的提取试剂、提取条件、色谱条件等进行了研究，以期建立黑果枸杞鲜果中花青素成分及含量测定的高效液相色谱法，为黑果枸杞鲜果中花青素的检测提供技术支撑，也为黑果枸杞鲜果及下游产品的质量标准检测体系提供科学依据。