HPLC Determination of α-keto Acids from Human Serum Using 2,3-Diamino-2,3-dimethylbutane as Derivatizing Reagent

  • Khalida Parveen Mahar Department of Chemistry, Faculty of Science, Shah Abdul Latif University Khairpur, Pakistan
  • Muhammad Yar Khuhawar Institute of Advanced Research Studies in Chemical Science, University of Sindh, Jamshoro, Pakistan
  • Kulsoom Ubedullah Abbasi Institute of Advanced Research Studies in Chemical Science, University of Sindh, Jamshoro, Pakistan
  • Gulam Qadir Shar Department of Chemistry, Faculty of Science, Shah Abdul Latif University Khairpur, Pakistan
  • Rafia Azmat Department of Chemistry, University of Karachi, Pakistan
  • Jameel Ahmed Baig National Centre of Excellence in Analytical Chemistry, University of Sindh, Jamshoro, Pakistan

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), 2oxoglutaric acid (KG) and Phenyl pyruvic acid (PPY) as derivatives of 2,3-diamino-2,3dimethybutane (DDB) were separated by HPLC column Zorbax C–18 (4.6x150 mm-id). The compounds were eluted with methanol-water-acetonitrile (40:58:2 V/V/V) with flow rate 1 ml/min. UV detection was carried out by photodiode array at 255 nm. Linear calibration plots were 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 3.249.71 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%. 

Published
Jun 3, 2013
How to Cite
MAHAR, Khalida Parveen et al. 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, [S.l.], v. 14, n. 2, p. 8, june 2013. ISSN 2221-5255. Available at: <http://pjaec.pk/index.php/pjaec/article/view/226>. Date accessed: 17 dec. 2017.