Pailla K, Blonde-Cynober F, Aussel C, De Bandt J P, Cynober L
Biochem Laboratory, Emile Roux Hospital, Limeil-Brévannes, France.
Clin Chem. 2000 Jun;46(6 Pt 1):848-53.
Measurement of keto-acids is important in various clinical situations. The aim of the present work was to develop a rapid HPLC method for the determination of keto-acids in human serum and to assess the concentrations of these acids in young adults and institutionalized elderly adults. This method was applied to the determination of blood keto-acid concentrations of young adults and institutionalized elderly people, divided into age groups
Four keto-acids (alpha-ketoisocaproate, alpha-ketoisovalerate, alpha-keto-beta-methylvalerate, and pyruvate) were derivatized with o-phenylenediamine to give fluorescent derivatives. After the sample preparation step (75 min to prepare 20 samples), the derivatives were separated chromatographically on a reversed-phase column using a binary gradient.
The fluorometric detection of the four keto-acids was rapid, <12 min. The method is repeatable and reproducible: the CVs were <6% and <11%, respectively, for each of the keto-acids. We found no significant difference between males and females. Concentrations of the branched-chain keto-acids decreased after age 60 years, especially alpha-ketoisocaproate, which decreased approximately 40%.
The proposed method allows rapid and reliable measurement of keto-acids. The data demonstrate that changes in branched-chain keto-acids concentrations in serum occur with age.
酮酸的测定在多种临床情况下都很重要。本研究的目的是开发一种快速高效液相色谱法（HPLC）用于测定人血清中的酮酸，并评估年轻成年人和机构养老老年人中这些酸的浓度。该方法应用于按年龄分组的年轻成年人和机构养老老年人血酮酸浓度的测定。
四种酮酸（α-酮异己酸、α-酮异戊酸、α-酮-β-甲基戊酸和丙酮酸）与邻苯二胺衍生化生成荧光衍生物。在样品制备步骤（制备20个样品需75分钟）后，使用二元梯度在反相柱上对衍生物进行色谱分离。
四种酮酸的荧光检测快速，<12分钟。该方法具有可重复性和再现性：每种酮酸的变异系数（CV）分别<6%和<11%。我们发现男性和女性之间无显著差异。60岁以后支链酮酸浓度下降，尤其是α-酮异己酸，下降了约40%。
所提出的方法能够快速可靠地测定酮酸。数据表明血清中支链酮酸浓度随年龄发生变化。
