HPLC法测定仔猪小肠刷状缘膜囊孵育体系中的二肽含量  

Determination of Dipeptide Content in Piglet Small Intestine Brush Border Membrane Incubation System with HPLC

作　　者：代建国[1] 李德发[1] 朴香淑[1] 杨文军[1] 武玉波[1] 

机构地区：[1]中国农业大学动物科技学院,北京100094

出　　处：《中国农业大学学报》2002年第3期94-98,共5页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目 (39870 583)

摘　　要：用高效液相色谱法测定了经仔猪小肠刷状缘膜囊水解后的甘 - L-脯、L-亮 -甘和 β-丙 -组共 3种二肽的剩余含量。分别以水 -三氟乙酸 (质量浓度分别为 0 .1 % ,0 .0 8%和 0 .1 % ,p H值分别为 2 .2 0 ,2 .30和 2 .2 0 )和乙腈 -三氟乙酸 (三氟乙酸质量浓度 0 .1 % )为流动相 ,经 Phenomennex ODS分析柱 (2 50 mm× 4.60 mm,5μm,C1 8)分离 ,在 2 0 0～ 40 0 nm处检测。结果表明 ,3种二肽在 0～ 30 0 μg.m L-1之间 ,其峰面积 (X)对质量浓度 (Y,μg.m L-1 )回归得方程分别为 Y=0 .0 0 0 1 X- 5.6945,(R2 =0 .9946) ;Y=0 .0 0 0 4 X- 1 .41 6 7,(R2 =0 .9999) ;Y=0 .1 842 X- 7.365 4,(R2 =0 .9948)。平均回收率依次为 1 0 0 .0 0 % ,1 0 0 .0 1 %和 99.92 % ,保留时间依次为 7.63,8.0 1和 4.1 7min,最大吸收波长依次为 2 0 1 ,2 0 0和 2 80 nm。该方法快速、简便和准确 ,结果稳定 ,重现性好 。The concentrations of Gly L pro, L Leu gly and β Ala his in piglet small intestine brush border membrane incubation system were analyzed by using high performance liquid chromatography. Water trifluoroacetic acid (concentrates were 0.1%, 0.08% and 0 1% respectively, pH value were 2.20, 2.30 and 2.20 respectively) and trifluoroacetic acid acetonitrile(concentrate was 0.1%) were used as mobile phase, sample were separated by Phenomennex ODS analytical column (250mm×4.60mm,5μm,C 18 ) and detected between 200～400nm. The results indicated that the regression equations of peak area ( X ) against concentrate ( Y ,μg·mL -1 )of the three dipeptides were, between 0～300μg·mL -1 , Y =0.0001 X -5.6945, R 2=0.9946; Y =0.0004 X -1.4167, R 2=0.9999 and Y =0.1842 X -7 3654, R 2=0.9948 respectively, the average recovery were 100 00%, 100.01% and 99.92% respectively. The determination method is rapid, simple, accurate, stable and reproducible, and is proposed to be used for determine content of the single and known dipeptide in piglet small intestine brush border membrane vesicle incubation system.

关 键 词：HPLC法 仔猪 小肠 刷状缘膜囊 孵育体系 二肽 含量 高效液相色谱法 甘-L-脯二肽 L-亮-甘二肽 β-丙-组二肽 

分 类 号：S828[农业科学—畜牧学] O657.72[农业科学—畜牧兽医]