反相高效液相色谱法测定烧伤大鼠体内的肠三叶因子

 采用反相高效液相色谱法测定烧伤大鼠体内的肠三叶因子 (ITF) ,其中色谱柱为HypersilC4柱 ( 4 6mmi.d .× 2 5 0mm ,柱填料的孔径为 3 0nm ,粒度为 5 μm) ,流动相为乙腈 水 (含体积分数为 0 1%的三氟乙酸 )溶液 ,梯度洗脱 ,流速为 1mL/min。在 2 14nm检测波长下 ,ITF在 1mg/L～ 10 0mg/L时具有良好的线性关系 ,相关系数为0 9989,最低检测限为 0 5mg/L。ITF的加标回收率为 93 95 %～ 10 5 90 % ,日内测定结果的RSD <5 3 3 % ,日间测定结果的RSD <6 10 %。方法准确、灵敏 ,可作为一种有效的分析手段。

Determination of Intestinal Trefoil Factor in Burned Rats by Reversed-Phase High Performance Liquid Chromatography

A reversed-phase high performance liquid chromatographic method was established for determination of intestinal trefoil factor (ITF) in burned rats. The analysis was carried out on a Hypersil C4 column (30 nm, 5 microns, 4.6 mm i.d. x 250 mm) with gradient elution of acetonitrile-trifluoroacetic acid-water solution at a flow rate of 1 mL/min and detected by a UV detector at 214 nm. The volume proportion of trifluoroacetic acid in water is 0.1% and the volume proportion of acetonitrile in mobile phase ranged from 19.5% to 42.0%. The linear range of the method was 1 mg/L-100 mg/L with relative coefficient of 0.9989. The minimum detection limit was 0.5 mg/L. The recovery of ITF added ranged between 93.95% and 105.90%. The intra-day and inter-day RSD of ITF were less than 5.33% and 6.10% respectively. This method is precise, accurate and can be used for the determination of ITF in intestinal mucosal of burned rats. Therefore, it is helpful in the research of comparing the effects of enteral feeding and parenteral nutrition on ITF expression.