Preparation of water-soluble,dyed substrates for amylase assay
Abstract
Abstract WATER-SOLBULE, DYED SUBSTRATES FOR USE IN ASSAYING AMYLASE ACTIVITY IN SUCH MEDIA AS BODY FLUIDS, PLANT EXTRACTS, AND THE LIKE, ARE PREPARED BY COUPLING, IN AN AQUEOUS ALKALINE SOLUTION, A REACTIVE DYE WITH A STRACH OR STRACH FRACTION, SUCH AS AMYLOSE OR AMYLOPECTIN AND REMOVING ALL UNREACTED DYE FROM THE WATER-SOLUBLE SUBSTRATE OBTAINED. GEL FILTRATION OR DIALYSIS CA BE USED AS THE PURIFICATION PROCESS, PROVIDED THE PROCESS IS MONITORED TO DETECT THE PRESENCE OF FREE DYE IN THE PURIFIED SOLUTION OF DYED SUBSTRATE. TO MONITOR THE PURIFICATION PROCEDURE, AN ALCOHOLIC TANNIC ACID PRECIPTATING AGENT IS USED TO PRECIPITATE THE DYED SUBSTRATE FROM A SAMPLE OF THE SOLUTION BEING PURIFIED AND THE SUPERNATANT FLUID IS OBSERVED FOR THE PRESENCE OF COLOR; COLOR IN THE SUPERNATANT FLUID INDICATES THAT SOME FREE DYE REMAINS, AND THAT THE DYED SUBSTRATE SOLUTION REQUIRES FURTHER PURIFICATION. THE PREFERRED PRECIPITATING AGENT CONTAINS A 0.5% TO 2% TANNIC ACID IN 50% ALCOHOL, BUFFERED TO A PH OF 5.0 TO 5.5 AND BROUGHT TO A TEMPERATURE OF FROM ABOUT 20*C. TO ABOUT 30*C.
Inventors
- 👤 Arthur L Babson
Bibliographic Details
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