Ascorbic and dehydroascorbic acids simultaneously quantified in biological fluids by liquid chromatography with fluorescence detection, and comparison with a colorimetric assay.
Ascorbic and dehydroascorbic acids simultaneously quantified in biological fluids by liquid chromatography with fluorescence detection, and comparison with a colorimetric assay.
复制标题
通过液相色谱和荧光检测同时定量生物体液中的抗坏血酸和脱氢抗坏血酸，并与比色测定进行比较。
作者：
A. Lopez;M. Mayersohn
我们描述了一种分离和定量血浆和尿液中抗坏血酸(AA)和脱氢抗坏血酸(DHA)的高效液相色谱方法。我们使用带有离子对试剂的反相C18柱，通过柱后反应与4，5-二甲基邻苯二胺形成荧光衍生物(在激发波长和发射波长分别为365和440 nm处测量)来检测分析物。异抗坏血酸(IA)为内标。DHA、AA和IA的保留时间分别为5.6、15.5和19.9min。血浆AA浓度为8和20 mg/L时，日内变异系数分别为9%和7%。AA和DHA的检出限分别为10和4 ng。结果本方法与甲氧基苯胺比色法测定AA的结果具有较高的准确性。
We describe a "high-performance" liquid-chromatographic method for separating and quantifying ascorbic acid (AA) and dehydroascorbic acid (DHA) in plasma and urine. We used a reversed-phase C18 column with an ion-pair reagent and detected the analytes by post-column reaction with 4,5-dimethyl-o-phenylenediamine to form a fluorescent derivative (measured at excitation and emission wavelengths of 365 and 440 nm, respectively). Isoascorbic acid (IA) is the internal standard. Retention times for DHA, AA, and IA are 5.6, 15.5, and 19.9 min, respectively. Between-day CVs for AA in plasma in concentrations of 8 and 20 mg/L were 9% and 7%, respectively. The limit of detection is 10 and 4 ng for AA and DHA, respectively. Results by the present method and the methoxyaniline colorimetric method for AA are comparably accurate.