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建立一种简单、快速、稳定性好的超高效液相色谱法测定酵母菌发酵乳清粉后游离氨基酸的变化。样品经6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)柱前衍生后,以AccQ·TagEluentA稀释液、乙腈和超纯水为流动相,梯度洗脱,在10min内实现了17种游离氨基酸的完全分离,在10~500μmol/L范围内呈良好的线性关系,最低检出限在0.0012~0.0024μmol/mL,加标回收率在84.5%~101.8%。结果表明:在乳清粉中检测到12种氨基酸,含量较高的是谷氨酸(Glu)、半胱氨酸(Cys),经酿酒酵母单独发酵乳清后丝氨酸(Ser)和半胱氨酸(Cys)含量显著上升,其余氨基酸含量均降低,乳酸克鲁维酵母存在的情况下游离氨基酸的含量均降低,这为研究酵母菌发酵乳清粉后游离氨基酸的变化提供理论支持。
To establish a simple, rapid and stable ultra-high performance liquid chromatography for the determination of free amino acids in yeast fermented whey powder. Samples were pre-column derivatized with 6-aminoquinolyl-N-hydroxysuccinimide (AQC) and eluted with AccQ-TagEluentA diluted with acetonitrile and ultrapure water as mobile phase with a gradient of 10 min The complete separation of 17 kinds of free amino acids was achieved, with a good linear relationship in the range of 10 ~ 500μmol / L with the detection limit of 0.0012 ~ 0.0024μmol / mL and the recoveries of 84% -101.8%. The results showed that 12 kinds of amino acids were detected in whey powder, and glutamic acid (Cys), serine (Ser) and cysteamine (Cys) content increased significantly, the remaining amino acid content decreased, the content of free amino acids decreased in the presence of Kluyveromyces lactis, which provided theoretical support for the study of free amino acids changes after yeast fermented whey powder.