Bourgeois C F, Mainguy P R
Int J Vitam Nutr Res. 1975;45(1):70-84.
Ascorbic acid (ASC) is separated by percolating the extract solution through an anionic Sephadex column. After one or two washings with water, the vitamin is oxidized on the column by a p-benzoquinone solution to dehydroascorbic acid (DASC). This latter being actually no acid is eluted as it is formed. The DASC containing eluate is treated with a new colorimetric reagent: 4-Nitro-1,2-Phenylenediamine (NPD). After extraction of the excess reagent with ethyl acetate, the optical extinction is measured at 375 nm. DASC, if present in the extract solution, can be reduced to ASC by dimercaptopropanol prior to chromatography. The method is very specific. The rather simple chromatographic purification can be effected semi-automatically with series of 10 colums (or more).
通过将提取液通过阴离子葡聚糖凝胶柱来分离抗坏血酸（ASC）。用水洗涤一两次后，维生素在柱上被对苯醌溶液氧化为脱氢抗坏血酸（DASC）。由于后者实际上不是酸，所以它一形成就被洗脱。含有DASC的洗脱液用一种新的比色试剂：4-硝基-1,2-苯二胺（NPD）处理。用乙酸乙酯萃取过量试剂后，在375nm处测量吸光度。如果提取液中存在DASC，在色谱分析之前可用二巯基丙醇将其还原为ASC。该方法非常特异。相当简单的色谱纯化可以通过一系列10个柱（或更多）半自动进行。
