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Measurement of plasma α-ketoisocaproate concentrations and specific radioactivity by high-performance liquid chromatography

✍ Scribed by Steven Nissen; Carol Van Huysen; Morey W. Haymond

Publisher
Elsevier Science
Year
1981
Tongue
English
Weight
307 KB
Volume
110
Category
Article
ISSN
0003-2697

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✦ Synopsis

An isocratic high-performance liquid chromatographic technique for the measurement of the specific radioactivity and concentration of a-ketoisocaproate in plasma is described. Plasma proteins are precipitated by additions of acetone, the supematant is applied to a cation-exchange column, and the resulting eluate is injected into a Cl,, reverse-phase column. Analysis time is approximately 10 min. Quantitative recovery, specificity, and sensitivity of this method are described and make this system attractive for in vivo o-ketoisocaproate kinetic studies. Using this procedure, the apparent flux of cu-ketoisocaproate in postabsorptive dogs was determined during an infusion of a-[U-"Clketoisocaproate and averaged 2.8 + 0.41 pmol/kg-min. r To whom reprint requests should be addressed at Mayo Clinic, Rochester, Minn. 55901.

* Abbreviations used: KIC, cy-ketoisocaproate; hplc, high-performance liquid chromatography; BCAA, branched-chain amino acid; gc, gas chromatography.

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