Abstract
Seven α-keto acids, pyruvic acid (PYR), 2-oxobutyric acid (KB), 3-methyl-2-oxobutyric acid (MKBA), 3-methyl-2-oxovaleric acid (K3MVA), 4-methyl-2-oxovaleric acid (K4MVA), 2-oxoglutaric acid (KG) and Phenyl pyruvic acid (PPY) as derivatives of 2,3-diamino-2,3-dimethybutane (DDB) were separated by HPLC column Zorbax C–18 (4.6x150 mm-id). The compoundswere eluted with methanol-water-acetonitrile (40:58:2 V/V/V) with flow rate 1 ml/min. UVdetection was carried out by photodiode array at 255nm. Linear calibration plotswere obtained with 0.1 to 60 μg/ml with limits of detection (LoD) within 0.04-0.4 μg/ml. The method was applied for the analysis of α-keto acids from serum of diabetic patients with blood glucose level 430-458 mg/dl and healthy volunteers. The amounts of α-keto acids observed 3.24-9.71 μg/ml with RSD 1.1-1.9% in diabetic patients were higher than healthy volunteer’s 0.11-1.3 μg/ml with RSD 0.9-2.6%.
Khalida Parveen Mahar, Muhammad Yar Khuhawar, Kulsoom Ubedullah Abbasi, Gulam Qadir Shar, Rafia Azmat, Jameel Ahmed Baig. (2013) HPLC Determination of α-keto Acids from Human Serum Using 2,3-Diamino-2,3-dimethylbutane as Derivatizing Reagent, Pakistan Journal of Analytical & Environmental Chemistry, Volume 14, Issue 2.
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