Molisch's Test
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Molisch's test is a sensitive
chemical test In chemistry, a chemical test is a qualitative property, qualitative or Quantitative property, quantitative procedure designed to identify, quantify, or characterise a chemical compound or substituent, chemical group. Purposes Chemical testing m ...
, named after Austrian botanist
Hans Molisch Hans Molisch (6 December 1856, Brünn, Habsburg Moravia - 8 December 1937, Wien, Federal State of Austria, Austria) was a Czech people, Czech-Austrians, Austrian botanist. Molisch's test is named after him, it is a sensitive chemical test for th ...
, for the presence of
carbohydrate A carbohydrate () is a biomolecule composed of carbon (C), hydrogen (H), and oxygen (O) atoms. The typical hydrogen-to-oxygen atomic ratio is 2:1, analogous to that of water, and is represented by the empirical formula (where ''m'' and ''n'' ...
s, based on the dehydration of the carbohydrate by sulfuric acid or
hydrochloric acid Hydrochloric acid, also known as muriatic acid or spirits of salt, is an aqueous solution of hydrogen chloride (HCl). It is a colorless solution with a distinctive pungency, pungent smell. It is classified as a acid strength, strong acid. It is ...
to produce an aldehyde, which condenses with two molecules of a phenol (usually
α-naphthol 1-Naphthol, or α-naphthol, is an organic compound with the formula . It is a fluorescence, fluorescent white solid. 1-Naphthol differs from its isomer 2-Naphthol, 2-naphthol by the location of the hydroxide, hydroxyl group on the naphthalene rin ...
, though other phenols such as
resorcinol Resorcinol (or resorcin) is a phenolic compound. It is an organic compound with the formula C6H4(OH)2. It is one of three isomeric benzenediols, the 1,3-isomer (or ''meta- (chemistry), meta''-isomer). Resorcinol crystallizes from benzene as co ...
and
thymol Thymol (also known as 2-isopropyl-5-methylphenol, IPMP), , is a toxic monoterpenoid phenol derivative of ''p''-Cymene, isomeric with carvacrol. It occurs naturally in the oil of thyme, and it is extracted from ''Thymus vulgaris'' (common thy ...
also give colored products), resulting in a violet ring.


Procedure

The test solution is combined with a small amount of Molisch's reagent (
α-naphthol 1-Naphthol, or α-naphthol, is an organic compound with the formula . It is a fluorescence, fluorescent white solid. 1-Naphthol differs from its isomer 2-Naphthol, 2-naphthol by the location of the hydroxide, hydroxyl group on the naphthalene rin ...
dissolved in ethanol) in a
test tube A test tube, also known as a culture tube or sample tube, is a common piece of laboratory glassware consisting of a finger-like length of glass or clear plastic tubing, open at the top and closed at the bottom. Test tubes are usually placed in s ...
. After mixing, a small amount of concentrated
sulfuric acid Sulfuric acid (American spelling and the preferred IUPAC name) or sulphuric acid (English in the Commonwealth of Nations, Commonwealth spelling), known in antiquity as oil of vitriol, is a mineral acid composed of the elements sulfur, oxygen, ...
is slowly added down the sides of the sloping test-tube, without mixing, to form a layer. A positive reaction is indicated by appearance of a purple red ring at the interface between the acid and test layers.


Reaction

All carbohydrates –
monosaccharide Monosaccharides (from Greek '' monos'': single, '' sacchar'': sugar), also called simple sugars, are the simplest forms of sugar and the most basic units (monomers) from which all carbohydrates are built. Chemically, monosaccharides are polyhy ...
s,
disaccharide A disaccharide (also called a double sugar or ''biose'') is the sugar formed when two monosaccharides are joined by glycosidic linkage. Like monosaccharides, disaccharides are simple sugars soluble in water. Three common examples are sucrose, ...
s, and
polysaccharide Polysaccharides (), or polycarbohydrates, are the most abundant carbohydrates found in food. They are long-chain polymeric carbohydrates composed of monosaccharide units bound together by glycosidic linkages. This carbohydrate can react with wat ...
s (except
triose A triose is a monosaccharide, or simple sugar, containing three carbon atoms. There are only three possible trioses: the two enantiomers of glyceraldehyde, which are aldoses; and dihydroxyacetone, a ketose which is symmetrical and therefore ha ...
s and
tetrose In organic chemistry, a tetrose is a monosaccharide with 4 carbon atoms. They have either an aldehyde () functional group in position 1 (aldotetroses) or a ketone () group in position 2 (ketotetroses). File:DErythrose Fischer.svg , D-Erythrose Fi ...
s)– should give a positive reaction, and
nucleic acid Nucleic acids are large biomolecules that are crucial in all cells and viruses. They are composed of nucleotides, which are the monomer components: a pentose, 5-carbon sugar, a phosphate group and a nitrogenous base. The two main classes of nuclei ...
s and
glycoprotein Glycoproteins are proteins which contain oligosaccharide (sugar) chains covalently attached to amino acid side-chains. The carbohydrate is attached to the protein in a cotranslational or posttranslational modification. This process is known a ...
s also give a positive reaction, as all these compounds are eventually hydrolyzed to monosaccharides by strong mineral acids.
Pentose In chemistry, a pentose is a monosaccharide (simple sugar) with five carbon atoms. The chemical formula of many pentoses is , and their molecular weight is 150.13 g/mol.furfural Furfural is an organic compound with the formula C4H3OCHO. It is a colorless liquid, although commercial samples are often brown. It has an aldehyde group attached to the 2-position of furan. It is a product of the dehydration of sugars, as occu ...
, while
hexose In chemistry, a hexose is a monosaccharide (simple sugar) with six carbon atoms. The chemical formula for all hexoses is , and their molecular weight is 180.156 g/mol. Hexoses exist in two forms, open-chain or cyclic, that easily convert into ...
s are dehydrated to 5-
hydroxymethylfurfural Hydroxymethylfurfural (HMF), also known as 5-(hydroxymethyl)furfural, is an organic compound formed by the dehydration of reducing sugars. It is a white low-melting solid (although commercial samples are often yellow) which is highly soluble in b ...
. Either of these aldehydes, if present, will condense with two molecules of
α-naphthol 1-Naphthol, or α-naphthol, is an organic compound with the formula . It is a fluorescence, fluorescent white solid. 1-Naphthol differs from its isomer 2-Naphthol, 2-naphthol by the location of the hydroxide, hydroxyl group on the naphthalene rin ...
to form a purple-colored product, as illustrated below by the example of
glucose Glucose is a sugar with the Chemical formula#Molecular formula, molecular formula , which is often abbreviated as Glc. It is overall the most abundant monosaccharide, a subcategory of carbohydrates. It is mainly made by plants and most algae d ...
: :


See also

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Rapid furfural test The rapid furfural test is a chemical test used to distinguish between glucose and fructose. The rapid furfural test is similar to Molisch's test but uses concentrated hydrochloric acid instead of concentrated sulfuric acid and the solution is boil ...


References

{{Analytical reagents Biochemistry detection methods Carbohydrate methods