An enzymic “sweep” technique for the chromatographic resolution of 14C-labeled brain metabolites

✍ Scribed by Thomas E. Duffy; John J. O'Neill

• Publisher: Elsevier Science
• Year: 1968
• Tongue: English
• Weight: 552 KB
• Volume: 24
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(68)90068-7

✦ Synopsis

A convenient method for evaluating pathways of tissue metabolism is based upon the radioisotope enrichment of intermediate compounds in any hypothetical scheme, given a common labeled presursor (l-3). In addition, measurements of specific activity often afford valuable insight into deviations from steady state, fluctuations in pool size, and rates of turnover among the individual components. For the rapid separation and specific activity determination of a broad spectrum of radioactive intermediates, a combination of column chromatographic and microfluorometric techniques seemed promising. However, a frequent complication arises from the appearance of multiple overlapping activity peaks, eluted in the chromatographic scan. In the present study, an attempt was made to resolve this difficulty by systematic removal of interfering substances upon addition of appropriate enzymes. The versatility of this approach was tested on extracts of adult guinea pig brain.