A histochemical method able to detect vitamins D and to discriminate it from cholesterol and its esters.
A suitable histochemical method able to discriminate vitamins D and 7-dehydrocholesterol from cholesterol and its esters is proposed taking into account the alkaline permanganate-toluidine blue reaction, the alkaline permanganate-Schiff reaction and the inhibitory effect of the HgCl2-formalin fixative. This method appears specific according to the results provided by spot test. On tissue sections the alkaline permanganate-toluidine blue reaction is more sensitive and more specific than the alkaline permanganate-Schiff reaction. A previous ribonuclease treatment or the methylation effect improves the specificity of the alkaline permanganate-toluidine blue reaction. The 2.4-dinitrophenylhydrazine blockade must be used previously to the alkaline permanganate-Schiff reaction as an attempt to improve its specificity. The previous hydrolysis carried on by some suitable treatments seems very useful to detect masked vitamins D and 7-dehydrocholesterol. The oxidized products accountable for the reactivity of vitamins D and 7-dehydrocholesterol to the alkaline permanganate-toluidine blue and alkaline permanganate-Schiff reactions were discussed.
W A
WA
Hadler
S R
SR
Silveira
D000818:Animals; D002415:Cats; D002784:Cholesterol / Q000032:analysis*; D002788:Cholesterol Esters / Q000032:analysis*; D004285:Dogs; D006168:Guinea Pigs; D006652:Histological Techniques; D006801:Humans; D051379:Mice; D009893:Opossums; D051381:Rats; D012867:Skin / Q000166:cytology*; D013194:Staining and Labeling; D014807:Vitamin D / Q000032:analysis*
D002788:Cholesterol Esters; D014807:Vitamin D; D002784:Cholesterol