采用薄层色谱法和改良竞争性蛋白结合分析法测定血浆中1,25-二羟基维生素D3
Determination of 1,25-dihydroxyvitamin D3 in plasma using thin-layer chromatography and modified competitive protein binding assay.
作者信息
Justová V, Wildtová Z, Pacovský V
出版信息
J Chromatogr. 1984 May 4;290:107-12. doi: 10.1016/s0021-9673(01)93565-4.
A simplified assay for calcitriol in plasma samples, using high-performance thin-layer chromatography (HPTLC) followed by competitive protein binding assay ( CPBA ), is described. For quantitative separation of femtomol concentrations of calcitriol from other metabolites of vitamin D, HPTLC can be used instead of column chromatography on Sephadex LH-20 and HPLC on silica gel. Since the CPBA with duodenal cytosol from chickens as a binding protein and [3H]1,25-dihydroxycholecalciferol as radioligand is highly sensitive, detection limit 10 fmol per assay, the volume of plasma sample can be reduced to 1 ml.
本文描述了一种用于血浆样本中骨化三醇的简化检测方法，该方法采用高效薄层色谱法（HPTLC），随后进行竞争性蛋白结合测定（CPBA）。为了从维生素D的其他代谢产物中定量分离飞摩尔浓度的骨化三醇，可用HPTLC代替在葡聚糖凝胶LH - 20上的柱色谱法以及在硅胶上的HPLC法。由于以鸡十二指肠细胞溶质作为结合蛋白、[3H]1,25 - 二羟基胆钙化醇作为放射性配体的CPBA具有高度敏感性，每次检测的检测限为10飞摩尔，因此血浆样本的体积可减少至1毫升。
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