Detection of Phosphate Esters on Paper Chromatograms View Full Text


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Article Info

DATE

1953-03

AUTHORS

H. E. WADE, D. M. MORGAN

ABSTRACT

PHOSPHATE esters separated on paper chromatograms are usually detected by degrading them on the paper with acid to give ortho-phosphoric acid and detecting this by the reaction with molybdate and a reducing reagent. The procedure of Hanes and Isherwood1 is based upon this and is widely used. This technique suffers from two disadvantages. In the first place, the prolonged initial ‘digestion’ necessary to break down the more resistant esters frequently leaves the paper in a very fragile state; and secondly, further analyses cannot be carried out on the spot after treatment. In order to avoid these disadvantages the following technique was developed. It depends upon the fixation of ferric ions by the esters and the reaction of the free ferric ion with salicylsulphonic acid. If the paper is not strongly buffered it is sprayed with 0.1 per cent FeCl3.6H2O in 80 per cent alcohol, dried in air at room temperature and then sprayed with 1 per cent salicylsulphonic acid in 80 per cent alcohol. Upon drying, the phosphates appear as white spots on a pale mauve background, ortho-phosphate having a band of deeper mauve surrounding it. The colour formation occurs only when the pH of the residual moisture in the paper is about 1.5–2.5. This can be checked conveniently with thymol blue, since the colour formation occurs most satisfactorily when the paper just turns this indicator red. Further indications of unsatisfactory pH are that below this range no colour is formed, whereas above the range the colour is orange-yellow and definition of the spots is very poor. More... »

PAGES

529-530

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/171529a0

DOI

http://dx.doi.org/10.1038/171529a0

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1030983761

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/13054615


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