RPLC-FLD快速测定不同颜色大豆中4种生育酚方法的优化及应用  

Optimization and Application of RPLC-FLD Method for Rapid Determination of Four Tocopherols in Different Color Soybean

作　　者：贾鹏禹[1,3] 李霞 杨一鸣[2] 吕喜滨 杜吉到 Jia Pengyu;Li Xia;Yang Yiming;LüXibin;Du Jidao(College of Food Science,Heilongjiang Bayi Agricultural University,Daqing 163319;College of Agriculture,Heilongjiang Bayi Agricultural University,Daqing 163319;National Coarse Cereals Engineering Research Center,Daqing 163319;Research Center of Saline and Alkali Land Improvement Engineering Technology in Heilongjiang Province,Daqing 163319)

机构地区：[1]黑龙江八一农垦大学食品学院,大庆163319 [2]黑龙江八一农垦大学农学院,大庆163319 [3]国家杂粮工程技术研究中心,大庆163319 [4]黑龙江省盐碱地改良工程技术研究中心,大庆163319

出　　处：《中国粮油学报》2025年第1期202-210,共9页Journal of the Chinese Cereals and Oils Association

基　　金：国家“十四五”重点研发计划子课题项目(2021YFD1201603-3);黑龙江省普通本科高等学校创新人才培养计划项目(UNPYSCT-2018086);黑龙江省“双一流”新一轮建设学科协同创新成果建设项目(LJGXCG2022-111)。

摘　　要：为快速测定不同颜色大豆中生育酚含量,利用C30分离柱结合荧光检测器建立了反相高效液相色谱测定4种生育酚的方法,重点优化了流动相组成和方法构成要素,考察了流动相添加甲基叔丁基醚(MTBE)对色谱分离效能的影响,并对方法展开了应用。最终方法采用YMC-carotenoid C30色谱柱分离结合荧光检测器检测,流动相采用梯度洗脱模式,流速为0.90 mL/min,色谱柱温度为30℃,荧光检测器激发波长294 nm,发射波长328 nm,进样体积10.0μL;样品前处理采用超声波辅助甲醇提取;方法针对不同种皮种仁颜色大豆中4种生育酚进行测定和评价。结果表明,流动相添加甲基叔丁基醚对目标化合物洗脱有良好的调控作用,能够提高检测效率;4种生育酚组分在13 min内完成分离,目标组分在50.0~5000.0 ng/mL浓度之间线性关系良好,检出限在2.17~5.19 ng/mL之间;不同种皮种仁颜色大豆中4种生育酚和总生育酚含量差异显著,生育酚总量在155.22~216.69μg/g之间。实验证明所建方法灵敏准确,简便易行,测试效率高,能够快速完成不同颜色大豆中全部形态生育酚的测定,在实际应用中取得了良好效果,为大豆营养品质评价提供技术支撑。For rapid determination of four tocopherols in different color soybeans,a method of reversed-phase high performance liquid chromatography was established by using C30 separation column and fluorescence detector.The composition of mobile phase and method components were optimized,and the effect by adding methyl tert-butyl ether(MTBE)to mobile phase on chromatographic separation efficiency was investigated,and the method was applied in practice.The YMC-carotenoid C30 column was used for separation combined fluorescence detector in the final method.The flow rate of mobile phase was 0.90 mL/min with gradient elution mode,and the column temperature was 30℃.The excitation wavelength of fluorescence detector was 294 nm,and the emission wavelength was 328 nm.The injection volume was 10.0μL.The samples were pretreated by ultrasonic assisted methanol extraction.Four kinds of tocopherols in soybean with different seed coats and kernel colors were determined and evaluated.The results indicated that the addition of methyl tert-butyl ether had a good regulation on the elution of target compounds to improve the detection efficiency.The four tocopherol components were separated within 13 min.The linear relationship of different components was good in the range of 50.0~5000.0 ng/mL,and the detection limit was 2.17~5.19 ng/mL.The contents of the four tocopherols in soybeans showed significant difference between different seed coat and kernel color,and the total amount of tocopherols ranged from 155.22μg/g to 216.69μg/g.The experiment proved that the method was sensitive,accurate,simple and easy to operate.It could complete the determination of all tocopherols in different color soybeans with high test efficiency.The good results achieved in practical application could provide technical support for the evaluation of soybean quality.

关 键 词：高效液相色谱 生育酚 大豆 方法优化 

分 类 号：S515[农业科学—作物学]