EXPERIMENT 9 - CARBOHYDRATES Monosaccharides - carbohydrates not decomposable to simple sugars by hydrolysis - all free monosaccharide are reducing carbohydrates

Monosaccharides used: arabinose (aldopentose) component of complex plant polysaccharide called gums. Found in free state in wood of coniferous trees (evergreen, shrubs) glucose (aldohexose) grape sugar/dextrose (dextrorotatory) - found in grapes - blood sugar fructose (ketohexose) fruit sugar; sweetest galactose (aldohexose) brain sugar; found in cerbrosides Oligosaccharides - contain two or ten simple sugars which are linked by glycosidic bonds Maltose (glucose + glucose) = malt sugar Lactose (glucose + galactose) = milk sugar Sucrose (glucose + fructose) = table sugar; found in sugar cane, sugar beets and sugar maple Polysaccharides - more than ten monosaccharide units - linked by glycosidic bonds - those found in nature are either structural or nutrient function Polysaccharides used: starch: amylase not branched but helical with hollow core amylopectin highly branched - found mostly in plant products agar-agar structural polysaccharide found in red algae -complete hydrolysis yields Galactose dextrin partial hydrolysis product of starch - used as adhesive and binders gum Arabic dried water-soluble carbohydrates from acacia Senegal -carbohydrate polymer, complex and highly branched -central core is arabinose -used in pharmaceutical preparations as adhesive, thickening agent glycogen used as a storehouse (liver and muscles) cellulose (cotton) most abundant organic compound on earth -linear polymer made up of glucose -most convenient lab source is filter paper MOLISCH TEST color test for sugars, which condenses with alpha naphthol on thymol in presence of strong sulfuric acid, which converts sugar to furfural derivatives - distinguishes carbohydrates from non-carbohydrates Reagent: -naphthol or thymol - dehydrates pentoses to form furfural and dehydrates hexoses to form 5-hydroxymethylfurfural. Furfurals further react with naphthol present in the test to reagents to produce a purple product - Purple color at interphase indicates a positive result ANTHRONE TEST used for qualitative and quantitative estimation of polysaccharides as well as monosaccharides - Based on dehydration of monosaccharides to furfural derivatives

Furfurals react with anthrone to form a deep green to blue color

IODINE TEST used to differentiate helical polysaccharides from non-helical - Iodine yields a blue-black color in the presence of starch and dextrin; indicates a positive result - Glycogen reacts with reagent to give a brown/blue color - Iodine atoms can then fit into the helices BENEDICTS TEST for detecting presence of reducing sugars - Contains Cu2+ ions in alkaline soln with sodium nitrate - Used in clinical labs for testing urine - Alkaline soln of copper are reduced by sugars having a free aldehyde - All tests positive except for sucrose Positive result = brick red ppt (Cu2O) BARFOEDS TEST uses Cu2+ ions in slightly acidic medium - If time of heating is carefully controlled, disaccharides dont react while reducing Reactions: Reducing monosaccharides are oxidized by Cu2+ in solution to form a carboxylic acid and brick red ppt of Cu2O within 2-3min Positive test for reducing monosaccharides: formation of green, red, or yellow ppt - Maltose, lactose, and sucrose yield negative results SELIWANOFFS TEST uses HCl as dehydrating acid and resorcinol as condensation agent - Distinguishes aldoses from ketoses - When mixed with seliwanoffs reagent, ketopentoses and ketohexoses react within 2min to form cherry red ppt Reaction: dehydrates ketohexoses to form hydroxymethylfurfural Positive results: Fructose & Sucrose only ORCINOLS TEST (BIALS TEST) orcinol (5-methylresorcinol) in HCl with FeCl3 catalyst - Used to distinguish pentoses from hexoses - Pentoses are converted to furfural which form a blue or green soln Positive results: Arabinose & Gum Arabic hydrolysate OSAZONE TEST Reducing sugars may be differentiated from each other by osazones they form with phenylhydrazine - Osazones have characteristic crystallic forms Positive Results: All except sucrose RESULTS: Fructose 2min (formed ppt) Glucose 4 to 5min Arabinose 10min Galactose 15 to 19min Lactose forms ppt when cooled Maltose - forms ppt when cooled Sucrose none MUCIC ACID TEST (GALACTARIC ACID TEST) to detect galactose, lactose, and agar-agar - Formation of transparent crystals - Hydrophilic substances or amphiphilic

EXPERIMENT 10 LIPIDS Lipids naturally occurring molecules - Fats, waxes ACROLEIN TEST to detect the presence of glycerol Reagent: KHSO4 Positive results: glycerol & lecithin UNSATURATION TEST used to detect the presence of double bonds in lipids by adding Hubls solution (from more unsaturated) Olive oil>Oleic acid>Coconut oil>Stearic acid (to saturated) TEST FOR PHOSPHATE gives yellow ppt for phospholipids 3NH4 + PO4 (from lecithin) + 12MoO4 + 24H+ (NH4)3PO4(MoO3)12 (yellow ammoniumphosphomolybdate) EMULSIFICATION TEST test if solution can form an emulsion with other substances - Used to detect polar and nonpolar groups in bile and lecithin Negative Result: Cholesterol CARR-PRICE REACTION used to detect presence of vitamin A Reagent: SbCl3 in CHCl3/0C -blue to green to gray to pink solution - Cod liver oil contains very high levels of vitamin A and vitamin D -carotene a precursor to vitamin A - highly conjugated and lipophilic MODIFIED FURTER-MEYER TEST is used to detect the presence of tocopherols by giving a bronze-red soln - Only -tocopherol is recognized to meet human requirements

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EXPERIMENT 11 ELEMENTARY COMPOSITION OF PROTEINS Proteins macromolecules - Made of amino acids; are polymers - Typical proteins contain 200-30 amino acids but some are smaller Peptide smallest protein Casein a protein found mostly in milk products - Used independently as a binding agent - 80% of milk content Synonyms: Milk solids - Calcium caseinate is produced from skim milk by adding an acid to cause the proteins to coagulate, where it can be filtered to separate TEST FOR CARBON, HYDROGEN AND OXYGEN decomposition by combustion (incomplete) Casein + O2(g) C(s)(charring black residue) + H2O (droplets formed at sides of container) TEST FOR NITROGEN NH4(g) + OH2 NH4 + OH- (turning red litmus to blue) SODIUM FUSION decomposition of organic bound elements by converting into their ionic species (as sodium salts) S, F (in casein) SO4, PO4