The absorption spectra of visual purple solutions extracted by various means were measured with a sensitive photoelectric spectrophotometer and compared with the classical visual purple absorption spectrum.
Hardening the retinas in alum before extraction yielded visual purple solutions of much higher light transmission in the blue and violet, probably because of the removal of light-dispersing substances.
Re-extraction indicated that visual purple is more soluble in the extractive than are the other colored retinal components. However, the concentration of the extractive did not affect the color purity of the extraction but did influence the keeping power. This suggests a chemical combination between the extractive and visual purple.
The pH of the extractive affected the color purity of the resulting solution. Over the pH range from 5.5 to 10.0, the visual purple color purity was greatest at the low pH. Temperature during extraction was also effective, the color purity being greater the higher the temperature, up to 40°C.
Drying and subsequent re-dissolving of visual purple solutions extracted with digitalin freed the solution of some protein impurities and increased its keeping power. Dialysis against distilled water seemed to precipitate visual purple from solution irreversibly.
None of the treatments described improved the symmetry of the unbleached visual purple absorption spectrum sufficiently for it to resemble the classical absorption spectrum. Therefore it is very likely that the classical absorption spectrum is that of the light-sensitive group only and that the absorption spectra of our purest unbleached visual purple solutions represent the molecule as a whole.