高效液相色谱法与pH示差法测定紫荚豌豆花青素含量的比较  

Determination of Anthocyanin in Purple Pea by High Performance Liquid Chromatography and pH Differential Method

作　　者：刘芳芳 向成钢 张宏[1] 徐加莲 刘琼枝 赵凯[1] 杨正安[1] 韩曙[1] LIU Fangfang;XIANG Chenggang;ZHANG Hong;XU Jialian;LIU Qiongzhi;ZHAO Kai;YANG Zheng'an;HAN Shu(College of Landscape and Horticulture,Yunnan Agricultural University,Kunming 650201,China;College of Biological and Agricultural Sciences,Honghe University,Honghe 661199,China;Yulong Agriculture and Rural Affairs Bureau,Lijiang 674100,China)

机构地区：[1]云南农业大学园林园艺学院,云南昆明650201 [2]红河学院生物科学与农学学院,云南红河661199 [3]云南丽江市玉龙县农业农村局,云南丽江674100

出　　处：《食品科技》2022年第1期298-303,共6页Food Science and Technology

基　　金：云南省重大专项(2018BB020-021-022);第十四届学生科技创新创业行动基金项目(2020ZKX099);云南省教育厅科学研究基金研究生项目(2020Y180)。

摘　　要：目的:采用高效液相色谱法(High performance liquid chromatography,HLPC)测定不同豌豆品种的花青素单体及含量,并与pH示差法测定结果进行了比较,旨在研究紫荚豌豆花青素定量分析方法的差异及其花青素的种类及功能。方法:紫荚豌豆以乙醇和盐酸作为萃取剂,采用安捷伦1260高效液相色谱仪进行测定,以10%的甲酸溶液、10%的甲酸甲醇溶液为流动相进行梯度洗脱,在波长520 nm的条件下检测紫荚豌豆花青素含量。pH示差法使用紫外分光光度计,在缓冲液pH值为1.0和4.5,30 ℃水浴60 min,波长为520 nm条件下测定花青素总含量。结果:利用高效液相色谱法测定紫荚豌豆的花色苷含量和种类,紫荚豌豆中含有飞燕草色素和矢车菊色素这2种花色苷,其中飞燕草色素含量显著高于矢车菊色素。利用紫外分光光度计检测总花青素,T0183的花青素含量最高。结论:相较于pH示差法测定紫荚豌豆花色苷总含量的结果,高效液相色谱法的测定含量更低,两者间的差异可能与紫荚豌豆中花色苷的种类和含量、HPLC法和pH示差法的花色苷标准品的选择以及pH示差法仪器准确度较低等相关。文章采用高效液相色谱法和pH示差法相结合的方式进行紫荚豌豆花青素定量分析,提高了花青素检测的稳定性与准确性。Objective:In this study,high performance liquid chromatography method was used to determine the anthocyanin monomer and content of different pea varieties,and the results were compared with those determined by pH differential method,aiming to study the difference of quantitative analysis methods of anthocyanins in purple pea,as well as the types and functions of anthocyanins.Method:Purple pea was determined by high performance liquid chromatography with ethanol and hydrochloric acid as extraction agents.Using 10% formic acid aqueous solution and 10% formic acid methanol solution as mobile phases to detect the anthocyanin in the purple pea at the wavelength of 520 nm.The pH differential method was used to determine the anthocyanin content under the conditions of 520 nm wavelength,pH buffer of 1.0 and 4.5.Results:Among the anthocyanin monomers of 6 kinds of purple peas determined by HPLC,delphinidin and cyanidin were detected in purple pea,in which delphinidin pigment was significantly higher than cyanidin.T0183 has the highest anthocyanin content in total anthocyanins detected by pH differential method.Conclusion:The results showed that the difference between HPLC and pH differential methods was related to the anthocyanins in purple pea,the selection of anthocyanins standard materials by HPLC and pH differential methods,and the low accuracy of pH differential method.In this paper,HPLC and pH differential method were used to quantitatively analyze anthocyanins in purple pea,which improves the stability and accuracy of anthocyanins detection.

关 键 词：紫荚豌豆 花色苷 高效液相色谱法(HPLC法) PH示差法 

分 类 号：TS207.3[轻工技术与工程—食品科学]