A new modified micromethod for quantitative estimation of cytochrome c

✍ Scribed by M. Osanai

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 562 KB
• Volume: 81
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90697-2

✦ Synopsis

A new convenient micromethod is presented for the quantitative determination of cytochrome c by measurement in the a-region of a low-temperature difference spectrum (reduced minus oxidized) using a commercially available split-beam spectrophotometer and its low-temperature spectrometry attachment. The method shows excellent reproducibility and precision with high sensitivity.

Interference by other pigments such as cytochrome b and b, was avoided by the extraction of cytochrome c from the material in 0.5% aluminium sulfate solution, pH 4.5. Direct extraction from the homogenate minimized the loss of cytochrome c. Distortion of the spectrum induced by low-temperature treatments was compensated for by calculations. With the present method, one can well estimate quantitatively 0.2 nmol of cytochrome c in a minimum of 0.8 ml (0.4 ml x 2) of sample. e.g.. the supematant obtained from five adults of Tenebrio molitor (500 mg: 2 ml of homogenate).