A specific method of high sensitivity for the determination of adenosine in the incubation medium of fat tissue.
A suitable method for the measurement of adenosine in the incubation medium of fat tissue (200-500 mg) has been developed. The method is based on the specificity of the adenosine deaminase reaction and on the high sensitivity of a fluorescent method for adenine derivatives. The decrease of fluorescence in a sample after treatment with this enzyme is used for measuring adenosine in the range of 50-500 pmoles/tube. This method is highly specific and is not affected by other adenine derivatives present in the sample. Instead of acetic acid, perchloric acid was used in the fluorescent reaction, thus increasing the amount of adenosine dependent fluorescence. With this modification of the original fluorescent method, perchloric acid extracts can be used without further processing after deproteinization of the samples. Using this method, we could measure the adenosine release of fat pads of Wistar rats incubated in Krebs-Ringer-albumin buffer without concentration or purification procedures.
D000241:Adenosine / Q000032:analysis*; D000243:Adenosine Deaminase; D000273:Adipose Tissue / Q000032:analysis*; D000818:Animals; D004822:Epididymis; D008297:Male; D010472:Perchlorates; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods
D010472:Perchlorates; D000243:Adenosine Deaminase; D000241:Adenosine