首页|HPLC同时检测发芽黑小麦中10种黄酮类化合物方法的建立

HPLC同时检测发芽黑小麦中10种黄酮类化合物方法的建立

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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(high-performance 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×5 μm)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20~100 µg/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≥0.999 1,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.
Establishment of an HPLC Method for Simultaneous Determination of Ten Flavonoids in Germinated Black Wheat
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×5 μm)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≥0.999 1,n=6)within the mass concentration range of 20~100 μg/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 acid-ethanol 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.

black wheathigh-performance liquid chromatographyflavonoidsgermination

李倩、张悦瑶、杨子怡、刘瑞、李小东、于章龙、孙元琳

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山西农业大学食品科学与工程学院,山西 太谷 030800

运城学院生命科学系,山西 运城 044000

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

山西农业大学棉花研究所,山西 运城 044000

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黑小麦 高效液相色谱 黄酮类化合物 发芽

2024

食品与生物技术学报
江南大学

食品与生物技术学报

CSTPCD北大核心
影响因子:0.674
ISSN:1673-1689
年,卷(期):2024.43(10)