HPLC同时检测发芽黑小麦中10种黄酮类化合物方法的建立  

Establishment of an HPLC Method for Simultaneous Determination of Ten Flavonoids in Germinated Black Wheat

作　　者：李倩 张悦瑶 杨子怡 刘瑞 李小东[1,4] 于章龙 孙元琳[2,3] LI Qian;ZHANG Yueyao;YANG Ziyi;LIU Rui;Li Xiaodong;YU Zhanglong;SUN Yuanlin(College of Food Science and Engineering,Shanxi Agricultural University,Taigu 030800,China;Department of Life Science,Yuncheng University,Yuncheng 044000,China;Shanxi Technology Innovation Center of High Value-Added Echelon Utilization of Premium Agro-Products,Yuncheng 044000,China;Institute of Cotton Research,Shanxi Agricultural University,Yuncheng 044000,China)

机构地区：[1]山西农业大学食品科学与工程学院,山西太谷030800 [2]运城学院生命科学系,山西运城044000 [3]山西省特优农产品梯次高值利用技术创新中心,山西运城044000 [4]山西农业大学棉花研究所,山西运城044000

出　　处：《食品与生物技术学报》2024年第10期109-119,共11页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(32072222);山西省基础研究计划项目(20210302124069);山西省现代农业产业技术体系建设专项资金(2024CYJSTX10-22);山西省研究生教育教学改革课题(2022YJJG269);运城市基础研究计划项目(YCKJ-2024024);运城学院食品科学与工程校级重点学科经费资助项目(XK-2021008);特色农产品发展学科群科研项目(SKX-202211)。

摘　　要：为探究黑小麦在发芽过程中黄酮类化合物质量分数的变化,建立了一种高效液相色谱(highperformance liquid chromatography,HPLC)同时检测发芽黑小麦中10种黄酮类化合物的方法。以Agilent 1260 ZORBAX SB-C18色谱柱(4.6 mm×250 mm×5μm)为固定相,乙腈和体积分数0.8%的磷酸水溶液为流动相进行梯度洗脱,流量设定为0.8 mL/min,检测波长为225 nm,柱温保持在25℃。结果表明,在质量浓度为20~100μg/mL时,葛根素、表儿茶素、芦丁、木犀草素、槲皮素、芹菜素、柚皮素、山柰酚、异鼠李素和查耳酮10种黄酮类化合物的质量浓度与峰面积之间呈现出良好的线性关系(R2≥0.9991,n=6);这些化合物的精密度、重复性和稳定性结果显示,峰面积相对标准偏差(relative standard deviation,RSD)均小于2.00%,加标回收率为98.72%~102.03%,表明所建立的检测方法准确可靠。另外,对发芽黑小麦中10种黄酮类化合物提取方法进行了优化,即采用乙醇体积分数为70%的盐酸-乙醇溶液(料液比1 g∶25 mL),在85℃下回流90 min时提取效果最佳,并检测了不同发芽时间(1、3、5、7 d)黑小麦中10种黄酮类化合物的质量分数。该研究为发芽黑小麦中功能活性成分的研究奠定了基础,同时也为其产品的质量控制和评估提供了参考依据。To investigate the changes in the mass fraction of flavonoids in the germination process of black wheat,a high-performance liquid chromatography(HPLC)method was established for simultaneous determination of 10 flavonoids in germinated black wheat.An Agilent 1260 ZORBAX SB-C18 column(4.6 mm×250 mm×5μm)was used as the stationary phase,with a gradient elution of acetonitrile and 0.8%phosphoric acid aqueous solution as the mobile phase.The flow rate was set at 0.8 mL/min,with a detection wavelength of 225 nm and the column temperature maintained at 25℃.The results showed that the mass concentration of 10 flavonoids,including puerarin,epicatechin,rutin,luteolin,quercetin,apigenin,naringenin,kaempferol,isorhamnetin,and chalcone,exhibited a good linear relationship with peak area(R2≥0.9991,n=6)within the mass concentration range of 20~100μg/mL.Precision,repeatability,and stability tests of these compounds showed that the relative standard deviation(RSD)of the peak area was less than 2.00%,and the spiked recovery rate ranged from 98.72%to 102.03%,demonstrating the accuracy and reliability of the established method.Additionally,the extraction method for 10 flavonoids in germinated black wheat was optimized,with the best efficiency achieved using a hydrochloric acidethanol solution with a 70%ethanol volume fraction(a solid-to-liquid ratio of 1 g∶25 mL),and refluxing at 85℃for 90 min.The mass fractions of 10 flavonoid were also determined in black wheat germinated for 1,3,5,and 7 d.This research lays a foundation for studying functional bioactive ingredients in germinated black wheat and offers a reference for the quality control and evaluation of its products.

关 键 词：黑小麦 高效液相色谱 黄酮类化合物 发芽 

分 类 号：O657.72[理学—分析化学] TS210.7[理学—化学]