TALLO, Kesha Marie D.

BMLS 2F

April 17, 2012

Qualitative test to detect CHO can be based on three basic properties: 1. Reducing property of the aldehyde and ketone group. 2. Hydroxyl groups can be dehydrated by strong acid and then treating the result with furfuran with several reagents to form a colored product. 3. Those which are highly pure, so a preparation of their derivatives could be performed followed by detection of CHO using various reagents.

QUALITATIVE TESTS FOR CARBOHYDRATES 1. Molisch Test This is a common test for all carbohydrates larger than tetroses. The test is on the basis that pentoses and hexoses are dehydrated by conc. sulphuric acid to form furfural or hydroxymethylfurfural, respectively. These products condense with -naphthol to form purple condensation product. 2. Benedicts Test - Free aldehyde or keto group in the reducing sugars reduce cupric hydroxide in alkaline medium to red colored cuprous oxide. Depending on the concentration of sugars, yellow to green color is developed . All monosaccharides are reducing sugars as they all have a free reactive carbonyl group. Some disaccharides like maltose have exposed carbonyl groups and are also reducing sugars but less reactive than monosaccharides. 3. Fehlings Test - This forms the reduction test of carbohydrates. Fehlings solution contains blue alkaline cupric hydroxide solution, heated with reducing sugars gets reduced to yellow or red cuprous oxide and is precipitated. Hence, formation of the yellow or brownish-red colored precipitate helps in the detection of reducing sugars in the test solution.

4. Seliwanoffs Test - It is a color reaction specific for ketoses. When conc. HCl is added, ketoses undergo dehydration to yield furfural derivatives more rapidly than aldoses. These derivatives form complexes with resorcinol to yield deep red color. The test reagent causes the dehydration of ketohexoses to form 5-hydroxymethylfurfural (HMF). 5-hydroxymethylfurfural reacts with resorcinol present in the test reagent to produce a red product within two minutes. Aldohexoses reacts so more slowly to form the same product. This test is used to distinguish an aldohexose form ketohexose.

5. Bials Test- Bials test is used to distinguish between pentoses and hexoses. They react with bials reagent and is converted to furfural. Orcinol and furfural condense in the presence of ferric ion to form a colored product. Appearance of green colour or precipitate indicates the

presence of pentoses and formation of muddy brown precipitate shows the presence of hexoses.

6. Iodine Test - This test is used for the detection of starch in the solution. The blue black colour is due to the formation of starch-iodine complex. Starch contain polymer of -amylose and amylopectin which forms a complex with iodine to give the blue black colour.

7.

Osazone Test - The ketoses and aldoses react with phenylhydrazine to produce a phenylhydrazone which further reacts with another two molecules of phenylhydrazine to yield the osazone. Needle-shaped yellow osazone crystals are produced by Glucose, fructose and mannose whereas lactosazone produces mushroom shaped crystals. Crystals of different shapes will be shown by different osazones. Flower-shaped crystals are produced by maltose.

Sources: http://amrita.vlab.co.in/?sub=3&brch=63&sim=631&cnt=1 http://www.scribd.com/doc/23452418/Qualitative-Test-for-Carbohydrates

TALLO, Kesha Marie D. BMLS 2F QUALITATIVE TEST FOR PROTEINS

April 17, 2012

1. Biuret Test - Biuret test is used for detecting the presence of peptide bonds. It relies on the reduction of copper (II)ions to copper(I), the latter form a complex with the nitrogens of the peptide bonds in an alkaline solution. A violet color indicates the presence of proteins. 2. Xanthoproteic Test Some amino acids contain aromatic groups that are derivatives of benzene. These aromatic groups can undergo reactions that are characteristics of benzene and benzene derivatives. One such reaction is the nitration of a benzene ring with nitric acid. The amino acids that have activated benzene ring can readily undergo nitration. This nitration reaction, in the presence of activated benzene ring, forms yellow product. 3. Ninhydrin Test - Ninhydrin (triketohydrindene hydrate) is a chemical used to detect ammoniaor primary and secondary amines. Amino acids also react with ninhydrin at pH=4. The reduction product obtained from ninhydrin then reacts with NH3 and excess ninhydrin to yield a blue colored substance. This reaction provides an extremely sensitive test for amino acids. 4. Millons Test - Millons test is specific to phenol containing structures (tyrosine is the only common phenolic amino acid). Millons reagent is concentrated HNO3, in which mercury is dissolved. As a result of the reaction a red precipitate or a red solution is considered as positive test. A yellow precipitate of HgO is NOT a positive reaction but usually indicates that the solution is too alkaline. 5. Hopkins Cole Test - The indole group of tryptophan reacts with glyoxylic acid (glacial acetic acid, which has been exposed to light, always contains glyoxylic acid CHOCOOH as an impurity) in the presence of concentrated H2SO4 to give a purple color. 6. Nitroprusside Test - The nitroprusside test is specific for cysteine, the only amino acid containing sulfhydryl group (-SH). This group reacts with nitroprusside in the presence of excess ammonia. 7. Sakaguchi Reaction - The Sakaguchi reagent is used to test for a certain amino acid and proteins. The amino acid that is detected in this test is arginine. Since arginine has a guanidine group in its side chain, it gives a red color with -naphthol in the presence of an oxidizing agent like bromine solution.
SOURCES: http://www.chem.boun.edu.tr/webpages/courses/Chem415/Chem%20415%20Experiment%202.pdf http://faculty.ksu.edu.sa/75115/BCH%20221Lectures/Enzyme%20lectures%20for%20Cat%201.pdf

TALLO, Kesha Marie D. BMLS 2F QUALITATIVE TEST FOR LIPIDS

April 17, 2012

1. Sudan Test - Sudan dyes have high affinity to fats, therefore they are used to demonstrate triglycerides, lipids, and lipoproteins. It uses the principle of the ability of fat cells to selectively absorb pigments in fat-dyes such as Sudan IV.

2. Acrolein Test - The principle behind the acrolein test is a specific chemical reaction. This reaction is utilized to determine the presence of glycerin in a fat. By heating the fat sample in the presence of potassium bisulfate (KHSO4), which acts as a dehydrating agent, acrolein (C3H4O, or CH2=CH-CHO) is formed and can easily be detected by its odor. Whenever fat is heated in the presence of a dehydrating agent, the fat molecule will shed its glycerol in the form of the unsaturated aldehyde - acrolein. SOURCES: http://wiki.answers.com/Q/What_is_the_principle_behind_the_acrolein_test http://en.wikipedia.org/wiki/Sudan_stain