Acidimetric assay for phospholipase A using egg yolk suspension as substrate

✍ Scribed by Nget-Hong Tan; Chon-Seng Tan

• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 555 KB
• Volume: 170
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(88)90632-x

✦ Synopsis

A convenient acidimetric assay for phospholipase A using egg yolk suspension as substrate has been developed. The substrate mixture consists of 1 part egg yolk, 1 part 8.1 mM sodium deoxycholate, and 1 part 18 mM calcium chloride. Phospholipase A activity is measured by following the initial rate of pH change, which is linear between pH 8.0 and 7.75 and is proportional to enzyme concentration over a wide range. The assay is highly reproducible, with a coefficient of variation of 3%, and as sensitive as most established assays for phospholipase A. The assay uses inexpensive and easily available substrate and is simple to perform. It is particularly useful for monitoring phospholipase A activity in chromatography fractions.