知识与财富的链接
【目的】为研究发芽对黑小麦中酚酸化合物质量分数的影响,建立了使用高效液相色谱法（HPLC）同时测定发芽黑小麦中11种酚酸化合物（没食子酸、原儿茶酸、对羟基苯甲酸、绿原酸、香草酸、咖啡酸、丁香酸、对香豆酸、阿魏酸、芥子酸和水杨酸）质量分数的方法。【方法】以甲醇（含体积分数1%乙酸）-水（含体积分数1%乙酸）为流动相,进行梯度洗脱,流量1 mL/min,检测波长280 nm,柱温25 ℃,对发芽第1、3、5、7天的黑小麦中11种酚酸化合物进行测定。【结果】方法学验证结果显示,在质量浓度为20~200 μg/mL时,11种酚酸化合物质量浓度与峰面积线性关系良好,相关系数（R2）≥0.999 4（n=6）,检出限（limit of detection,LOD）≥1.22 μg/mL,定量限（limit of quantification,LOQ）≥2.54 μg/mL,精密度、重复性和稳定性试验中11种酚酸化合物色谱峰的峰面积相对标准偏差（relative standard deviation,RSD）均小于2.00%,平均加标回收率为98.02%~101.12%,表明所建立的测定方法快捷,且结果准确可靠。在提取条件优化中发现,当料液比为1 g∶30 mL时,以体积分数为80%的甲醇溶液涡旋提取6 min,所测发芽黑小麦中11种酚酸化合物总质量分数较高。发芽黑小麦中可以检测出没食子酸,且在发芽第3天质量分数达到峰值;11种酚酸化合物总质量分数在发芽第5天最高,是未发芽黑小麦对照组的2.13倍,这表明发芽处理能改变黑小麦中酚酸化合物的质量分数。【结论】该研究评价了不同提取条件对发芽黑小麦中11种酚酸化合物的影响,并对其质量分数进行测定,可为发芽黑小麦中功能活性成分的研究奠定基础,也为其产品的质量控制和评估提供参考。
[Objective]In order to study the effects of germination on the mass fractions of phenolic acids in black wheat, a method was established for simultaneous determination of 11 phenolic acids (gallic acid, protocatechuic acid, 4-hydroxybenzoic acid, chlorogenic acid, vanillic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, sinapic acid, and salicylic acid) in germinated black wheat based on high performance liquid chromatography (HPLC).[Method]The mobile phase composed of methanol (containing acetic acid at a volume fraction of 1%) and water (containing acetic acid at a volume fraction of 1%) was used for gradient elution at a flow rate of 1 mL/min, a detection wavelength of 280 nm, and a column temperature of 25 ℃, to determine the mass fractions of 11 phenolic acids in black wheat on the 1st, 3rd, 5th and 7th days of germination.[Result]The results of methodology validation showed that 11 phenolic acids had a good linear relationship between mass concentration and peak area within the mass concentration range from 20 to 200 μg/mL, R2≥0.999 4 (n=6). The established method showed the limit of detection (LOD) ≥1.22 μg/mL and the limit of quantification (LOQ) ≥2.54 μg/mL for the phenolic acids. The relative standard deviations (RSD) of peak area of precision, repeatability, and stability were all less than 2.00%, and the average spiked recovery rates were between 98.02% and 101.12%, indicating that the established method was fast, accurate, and reliable. The extraction conditions were optimized as extraction with the solid-liquid ratio of 1 g∶30 mL and methanol at a volume fraction of 80% by vortex for 6 min, under which the total mass fraction of 11 phenolic acids in germinated black wheat was the highest. The results showed that during germination, the mass fraction of gallic acid in black wheat reached the peak on the 3rd day of germination. The total mass fraction of 11 phenolic acids was the highest on the 5th day of germination, which was 2.13 times of that in the non-germinated control group. This indicated that germination changed the mass fractions of phenolic acids in black wheat.[Conclusion]This study evaluated the effects of extraction conditions on 11 phenolic acids and determined the mass fractions of the 11 phenolic acids in germinated black wheat. The findings lay a foundation for the study of functional components in germinated black wheat and provide a reference for the quality control and evaluation of its products.
