TERPENOIDS AND STEROIDS 



163 



TABLE 14. LITERATURE REFERENCES TO PAPER 

 CHROMATOGRAPHY OF TERPENOIDS AND STEROLS 



Type of Compound 



Terpenoid acids 



Saponins 



Steroids 



Carotenoids 

 Cardiac glycosides 

 Ubiquinone derivatives 

 Sterols, terpenoid alcohols 



Author and Reference Number 



Pasich (69) 



Sannie et al. (70), Pasich (71) 



Neher (59), Reineke (72), Axelrod 

 and Pulliam (73) 



V 



Sestak (74), Jensert and Jensen (75) 

 Resplandy (76), Kowalewski (77) 

 Lester and Ramasarma (78) 

 Peereboom et al. (79) 



anhydride and a drop of concentrated sulfuric acid. The mechanism and specificity of 

 this test are discussed by Brieskorn and Herrig (85) and Cook (86). This, and some of 

 the other common color reactions, cannot be applied to paper strips because of their de- 

 structive effect on the paper but might be used on glass fiber paper or on silica coated 

 glass strips. The well-known Legal reaction for cardiac glycosides is given by many 

 substances containing the grouping CH-CO and is therefore of value only for testing sub- 

 stances already shown to be steroids. It is also not given by the scilladienolides. Penta- 

 cyclic triterpenoids give a violet color when heated with 2, 6-di-/er^-butyl-p-cresol in 

 ethanol. Steroids give no color or a yellow-green one (87). Gerlach (88) has reviewed 

 eleven well-known color reactions and recommends the reagent of Brieskorn and Briner 

 (chlorosulfonic acid and Sesolvan NK) for specifically distinguishing between triterpenoids 

 (red color) and steroids (brown color). Several color reactions for detecting steroids 

 and terpenoids on chromatograms have been reviewed by Wachsmuth and Koeckhoven(89). 

 Most saponins are readily recognized by their hemolytic property although some may be 

 quite weak in this respect. They can be added in isotonic solution to defibrinated or cit- 

 rated blood and hemolysis observed, or paper chromatograms may be sprayed with iso- 

 tonically diluted blood and hemolytic zones observed. Shamma (90) has presented a useful 

 review of diagnostic reactions which may be used to distinguish among the three basic 

 type structures of the pentacyclic triterpenoids. A color reaction relatively specific for 

 ubiquinone derivatives is based on their formation of a blue color with ethyl cyanoacetate 

 and ammonia (91). 



If pure compounds have been separated by chromatography or other means, deter- 

 mination of absorption spectra is of great value in assigning them to groups. In the ultra- 

 violet and visible region absorption of these compounds is mostly due to the presence of 

 conjugated double bond systems. Compounds with isolated double bonds have no absorp- 

 tion peaks in the visible or ultra-violet spectrum above 200 m^- 



Woodward (92) has presented rules for predicting the absorption maximum for a 

 diene system, taking the base value of 217 mii for a conjugated double bond and adding 

 appropriate increments for different structural features. Measurements are generally 

 in close agreement with predicted values. Ultraviolet and infrared spectra of over 200 

 sesquiterpenes are recorded by Pliva et al. (63). The carotenoids show strong absorp- 



