HPLC法分析动物血浆和组织中的谷氨酰胺

Determination of glutamine in plasma and tissues by high performance liquid phase chromatography

以邻苯二甲醛为衍生剂 ,柱前衍生 ,反相 C1 8柱分离 ;p H6.85磷酸缓冲溶液 ,甲醇二元梯度洗脱 ,3 40 nm紫外检测。结果显示 ,谷氨酰胺 (Gln)在 1 5 .7min出峰 ,在 0 .0 5 3 -1 .0 5 4mmol/L范围内与峰面积呈良好的线性关系 (r=0 .984)且P<0 .0 1 ,血浆、肝、肌肉和肾脏谷氨酰胺回收率在 94%～ 99%之间。方法应用中测定鸡血浆谷氨酰胺浓度是 0 .63± 0 .1 2mmol/L;谷氨酰胺在羊血浆、肝、肌肉和肾脏中的分别为 0 .42± 0 .0 73 mmol/L、1 .80± 0 .1 3 μmol/g、1 .84± 0 .1 1 μmol/g及 0 .74± 0 .1 1μmol/g(n=3 ) 。

A pre column deviation precedure of O phthaldialdelhyde( OPA ) was used for the determination of glutamine(Gln) by reversed phase high performance liquid chromatography. A phosphate buffer at pH 6 85 was used as the mobile phase A and methanol as B. Gradient elution was adopted with ultraviolet detector(λ=340 nm).The result shoes that the retention time of Gln is 15 7 min. A good linear relationship exits between peak area and Gln concentration ranging from 0 053 mmol/L to 1 054 mmol/L. The correlation coefficiet is 0 984 and P<0 01. The Gln recovery of plasma, liver, muscle and kidney is in the range of 95%~99%. In its application, Gln in chicken plasma is 0 63±0 12 mmol/L. The Gln concentrations of goat plasma, liver, muscle and kidney are 0 42±0 073 mmol/L, 1 80±0 13 μmol/g, 1 84±0 11 μmol/g and 0 74±0 11 μmol/g(n=3), respectively. Gln in liver and muscle are higher.