I'm doing extraction of saponin from banana peels. i use vanillin-sulfuric acid assay to but got some error. all my samples become black after incubation and give 3.00 absorbance. may i know what colour i should observe after adding sulfuric acid?
the procedure of the assay that i follow from literature review:
-1 ml of sample extract + 0.25 ml 8% (v/v) vanillin + 2.5 ml 72% H2SO4
-incubate mixture in 60oC water bath for 10 minutes, then in ice for 15 minutes
-record A544
*when i add acid, the mixture become yellow and darker over time, totally black after incubation
does anyone know any journals that i can refer to regarding this assay? i need to find out the function of vanillin, reaction of acid with my sample/saponin.
Thanks a lot!
Please find the detailed method of vanillin reaction and its principle in the attached article.
if the colure is changing then you may need to protect from light. but try the link below.
https://www.users.miamioh.edu/hagermae/Vanillin%20Assay.pdf
Thank you Mr. Amit and Mr. Amir, coincidentally both of you share the same article. But not much information I can use from there.
Dear Sabah Mabrouki, may i know what colour of the reaction do u observe? is the procedure similar to what i do?
In my assays the same thing happens...when I add the acid, the mixture becomes yellow. Did you find a explanation or solution?
Thanks
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