反相高效液相色谱法测定血浆和尿液中的菊粉
Determination of inulin in plasma and urine by reversed-phase high-performance liquid chromatography.
作者信息
Dall'Amico R, Montini G, Pisanello L, Piovesan G, Bottaro S, Cracco A T, Zacchello G, Zacchello F
机构信息
Department of Pediatrics, University of Padua, Italy.
出版信息
J Chromatogr B Biomed Appl. 1995 Oct 6;672(1):155-9. doi: 10.1016/0378-4347(95)00209-2.
We report a new HPLC procedure for measuring inulin in plasma and urine. Samples after dilution are boiled in mild acidic conditions and then analyzed on a C18 column. Solvent system A is 3.2 mM HCl, pH 2.5, and B is acetonitrile-3.2 mM HCl (60:40, v/v), pH 2.5. The separation is carried out in 8 min with a flow-rate of 1.0 ml/min and the absorbance monitored at 280 nm. The relationship between inulin and the recorded peak area is linear from 0.2 to 3.2 mg/ml with a correlation coefficient of 0.999 for plasma and 0.999 for urine. Within-run precision, measured at three inulin concentrations, ranged from 0.9 to 1.7% in plasma and from 0.8 to 1.2% in urine. Between-run precision varied in plasma from 2.7 to 3.2% and in urine from 3.0 to 3.3%. Analytical recovery ranged from 102 to 107% in plasma and from 101 to 105% in urine, respectively. The method is sensitive, selective and only 30-microliters samples are required. Therefore, it could be used to evaluate the glomerular filtration rate even in small babies and to perform studies in animals.
我们报告了一种用于测量血浆和尿液中菊粉的高效液相色谱新方法。稀释后的样品在温和酸性条件下煮沸，然后在C18柱上进行分析。溶剂系统A为3.2 mM盐酸，pH 2.5，溶剂系统B为乙腈 - 3.2 mM盐酸（60:40，v/v），pH 2.5。分离在8分钟内完成，流速为1.0 ml/min，在280 nm处监测吸光度。菊粉与记录的峰面积之间的关系在0.2至3.2 mg/ml范围内呈线性，血浆的相关系数为0.999，尿液的相关系数为0.999。在三种菊粉浓度下测量的批内精密度，血浆中为0.9%至1.7%，尿液中为0.8%至1.2%。批间精密度在血浆中为2.7%至3.2%，在尿液中为3.0%至3.3%。分析回收率在血浆中分别为102%至107%，在尿液中为101%至105%。该方法灵敏、具有选择性，仅需30微升样品。因此，它可用于评估甚至是小婴儿的肾小球滤过率，并可用于动物研究。
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