HPLC测定黑果枸杞鲜果中花青素含量的方法研究
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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。
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(R2of 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..
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(R2of 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..
引文
[1]彭祖茂,邓梦雅,严虞虞,等.植物中花青素含量测定及种类分布研究[J].食品研究与开发,2018,17(39):100-104.
[2]徐美玲,赵德卿.蓝莓花青素的提取及理化性质的研究[J].食品研究与开发,2008(09):187-189.
[3]陶大勇,陈家娟,陈瑛,等.黑果枸杞色素对小鼠抗衰老作用的研究[J].中兽医医药杂志,2008(1):11-13.
[4]徐春明,庞高阳,李婷.花青素的生理活性研究进展[J].中国食品添加剂,2013(3):205-210.
[5]赵子丹,葛谦,牛艳,等.HPLC测定黑果枸杞中花青素的成分及含量[J].食品研究与开发,2017,12(38):123-129.
[6]周荣朝,刘银声.数学研究应用于生产实践的尝试-正交试验在生物化学中的应用[J].华北水利水电学院学报,1995,3:23-26.
[2]徐美玲,赵德卿.蓝莓花青素的提取及理化性质的研究[J].食品研究与开发,2008(09):187-189.
[3]陶大勇,陈家娟,陈瑛,等.黑果枸杞色素对小鼠抗衰老作用的研究[J].中兽医医药杂志,2008(1):11-13.
[4]徐春明,庞高阳,李婷.花青素的生理活性研究进展[J].中国食品添加剂,2013(3):205-210.
[5]赵子丹,葛谦,牛艳,等.HPLC测定黑果枸杞中花青素的成分及含量[J].食品研究与开发,2017,12(38):123-129.
[6]周荣朝,刘银声.数学研究应用于生产实践的尝试-正交试验在生物化学中的应用[J].华北水利水电学院学报,1995,3:23-26.