1958 年 78 巻 4 号 p. 424-427
As a simple method for measuring the insulin content in solution, paper chromatography was adopted. In the case of a solution containing a large amount of impurities, such as the pancreatic extract, it is submitted to pretreatment of adjusting the pH so as to precipitate and remove the large amount of occluded proteins, desalted by dialysis with fish skin or cellophane membrane, concentrated under a reduced pressure by rotary evaporator, and a definite amount of the concentrated solution is submitted to paper partition chromatography. It is streaked at one end of a filter paper (40×40cm.), developed with a solvent system of acetic acid: butanol: water (1:6:7), and the insulin band separated on the papergram is colored by 0.02% solution of bromocresol green. The intensity of the color thereby developed is estimated by the measurement of the colored eluate of the insulin band by photoelectric colorimeter and insulin unit is calculated from the straight linear relationship between the concentration of the pigment and units of insulin.
This method was applied to the extract solution obtained in a plant using whale pancreas as the starting material and the content of insulin was found to be 1000-1500 I.U./kg. in fin whale and 2300 I.U./kg. in sperm whale.