Koike K, Koike M
Anal Biochem. 1984 Sep;141(2):481-7. doi: 10.1016/0003-2697(84)90074-5.
A procedure for rapid separation and microquantitative determination of various alpha-keto acids in serum and urine was developed. The procedure used reverse-phase high-performance liquid chromatography of the alpha-keto acids after derivatization into fluorescent quinoxalines by reaction with o-phenylenediamine. Deproteinization of serum with tungstic acid or methanol facilitated a constant recovery of alpha-keto acids. The useful range of analysis of seven alpha-keto acids by isocratic chromatography was from 10 to 250 pmol. The fluorescence emission was measured at 410 nm with excitation at 350 nm. The data obtained from samples of patients with chronic pyruvic acidemia and maple syrup urine disease, confirmed the usefulness of the method in clinical applications. A slightly modified procedure was needed for the analysis of oxaloacetic acid and phenylpyruvic acid.
开发了一种用于快速分离和微量定量测定血清和尿液中各种α-酮酸的方法。该方法采用反相高效液相色谱法，通过与邻苯二胺反应将α-酮酸衍生化为荧光喹喔啉后进行分析。用钨酸或甲醇对血清进行脱蛋白处理有助于α-酮酸的回收率恒定。通过等度色谱法分析七种α-酮酸的有效范围为10至250皮摩尔。在350nm激发波长下于410nm处测量荧光发射。从慢性丙酮酸血症和枫糖尿症患者样本中获得的数据证实了该方法在临床应用中的有效性。分析草酰乙酸和苯丙酮酸需要稍微修改一下方法。
