Schiff test
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The '''Schiff test''' invented <ref>''Eine neue Reihe organischen Diaminen'' H. Schiff Justus Liebigs Ann Chemie 140, 92-137 '''1866''' </ref> and named after [[Hugo Schiff]] is a [[chemical test]] for the detection of [[aldehydes]] <ref>''Histology, cell and tissue biology'' 5th Ed., 1983 ISBN 0333354060</ref>. An unknown sample is added to the decolorized '''Schiff reagent''' and when aldehyde is present a characteristic [[magenta]] or purple color develops. The Schiff reagent is the reaction product of [[Fuchsine]] or the closely related [[Pararosaniline]] (lacks a [[methyl]] group) and [[sodium bisulfite]]. Human skin also contains aldehydes and gets stained as well.
Schiff reagents are used for various [[staining]] methods, eg. [[Feulgen stain]] and [[periodic acid-Schiff stain]].
==Mechanism==
Fuchsin itself is colored due to its [[quinoid]] structure (see also for example [[viologen]]) but is discolorized by [[sulfonation]] at the central carbon atom which disrupts the favorable [[resonance structure|resonance]] in the molecule. More bisulphite (depending on stoichiometry ) reacts with available [[aniline|amine]] groups to so-called N-sulfinic acid groups.
The reaction of the Schiff reagent with aldehydes is complex with several research groups reporting multiple reaction products with model compounds. Two different mechanisms appear in the literature <ref>''Histochemistry, theoretical and applied'' 4th Ed. 1985 ISBN 044302997 Note: the depiction of the sulfonic acid mechanism in this edition contains an error as the aldehyde R group is bonded to nitrogen and not to its carbon neighbor</ref> The first mechanism explaining its action with aldehydes was proposed by H. Wieland in 1935. In it the aldehyde groups react with the sulfinic acid groups forming an [[sulfonamide]].
:[[Image:SchiffReagentMechanismI.png|400px|Schiff reagent sulfonamide mechanism]]
In the second mechanism, the pararosanilin, sulfurous acid and aldehyde combine to form an [[alkyl sulfonic acid]] with sulfur bonded to carbon rather than to nitrogen. This mechanism was first proposed by P. Rumpf in 1935 and experimental evidence was obtained in 1964 by Hardonk and van Duijn <ref>''THE MECHANISM OF THE SCHIFF REACTION AS STUDIED WITH HISTOCHEMICAL MODEL SYSTEMS'' M. J. HARDONK and P. VAN DUIJN J. Histochem. Cytochem., Oct '''1964'''; 12: 748 - 751. </ref>. They noted that when the aldehyde is reacted first with sulfurous acid and then with the Schiff compound the [[UV spectrum]] is identical to one obtained in a standard test. This particular logic was disputed by Stoward in 1966 although on the whole he agreed with the new mechanism <ref>''SOME COMMENTS ON THE MECHANISM OF THE SCHIFF REACTION'' P. J. STOWARD J. Histochem. Cytochem., Sep '''1966'''; 14: 681 - 683. </ref>. In 1980 [[NMR spectroscopy]] (Robins, Abrams, Pincock) provided further support for the sulfonic acid mechanism <ref>''The structure of Schiff reagent aldehyde adducts and the mechanism of the Schiff reaction as determined by nuclear magnetic resonance spectroscopy'' J.H. Robins, G.D. Abrams, and J.A. Pincock Can. J. Chem./Rev. can. chim. 58(4): 339-347 ('''1980''') </ref>.
:[[Image:SchiffReagentMechanismII.png|400px|Schiff test|Schiff reagent sulfonic acid mechanism]]
==External links ==
* [http://stainsfile.info/StainsFile/stain/schiff/schiffwhatis.htm What is Schiff's reagent?]
==References==
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[[category:chemical tests]]
[[de:Schiffsche Probe]]
[[es:Reactivo de Schiff]]
[[fr:Réaction de Schiff]]
[[pt:Reagente de Schiff]]