Carbohydrates Definition: Organic compounds composed of C, H and O with H and O present in the same ratio as in water. E.g. Glucose C6H12O6. Exceptions: Deoxy sugars such as Rhamnose C6H12O5, digitoxose C6H12O4 Some non carbohydrates follow the definition: Acetic acid C2H4O2 Formaldehyde HCHO Lactic acid C3H6O3 New definition: Optically active Polyhydroxy aldehydes or ketones, or substances that hydrolyze to yield polyhydroxy aldehydes or ketones. Carbohydrates Non-Sugars (Polysaccharides) Sugars (Saccharides) More than 10 sugar unites Monosaccharides Oligosaccharides (3- 10 unites) Disaccharides Trioses e.g. Glyceraldehyde Trisaccharides e.g. Raffinose Tetroses e.g. Erythose Pentoses e.g. Arabinose Hexoses Reducing Lactose Maltose Cellibiose Non-Reducing Sucrose Tetrasaccharides Pentasaccharides Aldohexeoses Glucose Ketohexeoses Fructose Non-Sugars (Polysaccharides) More than 10 sugar unites Homopolysaccharides Holosaccharides Hexosanes Glucosans Starch Glycogen Cellulose Dextrins Heteropolysaccharides Pentosans Xylans Fructosans Inulin Gums Mucilages Pectins Agar Heparin Alginate Physical Characters Condition: Sugars are white, crystalline in shape and with sharp melting points, while polysaccharides are white amorphous solids. Taste: Sugars have a sweet taste. Polysaccharides are tasteless. Solubility: Monosaccharides are soluble in cold water and hot alcohol. Polysaccharides are partially soluble in hot water. Optical activity: A compound is optically active when, in solution, it is capable to rotate the plane of polarized light either to right (dextrorotatory, + or d) or to the left (levorotatory, - or l). Nickel prism Plane polarized light Normal light The optical activity of a compound is measured by determination of its specific optical rotation ( Dt) using a polarimeter, and applying the following equation: D25 = /LC Where: = extension of rotation, L = length of tube (light path) in decimeter, C = concentration g /ml, 25 = operating temperature (t) in 0C, D = line spectrum of sodium light (589 nm). Sugar isomers: Hexoses like glucose have 4 asymmetric (chairal) carbons. Number of isomers can be calculated from the formula: Number of isomers = 2n = 24 = 16 CHO * CH OH * HO CH * CH OH * CH OH CH2OH D and L in sugars: A monosaccharide in which the OH group attached to the carbon atom next to the CH2OH (farthest asymmetric carbon atom from the carbonyl group) is always to the right is designated as a “D-sugar” and that with the same OH to the left as “L -sugar”. D and L designations are like (R) and (S) designations in that they are not necessarily related to the optical rotations of the sugars to which they are applied. Thus, one may encounter sugars that are D (+) or D (-) and others that are L (-) or L (+). CHO H CHO OH CH2OH D-Glyceraldehyde HO H CH2OH L-Glyceraldehyde - and - anomers of glucose: When sugars under go cyclization C-1 became a new chairal carbon and two isomers exist. They are called “ Anomers”. In the -anomer the OH group is directed downside and in the -anomer is directed to the upper side. These two forms have different specific rotation, in solution an equlibrium exsit between the two forms (mutarotation phenomenon). H OH H OH H O HO HO H H 1 H OH OH HO HO -glucopyranoside Chair forms 6 H O H H H 1 OH OH H -glucopyranoside CH 2OH CH 2OH O H OH H H H 1 OH OH H OH -glucopyranoside H OH O OH H 1 H OH H OH -glucopyranoside Haworth formulations Mutarotation When a sugar is dissolved in water, the specific rotation of the solution gradually changes until it reaches to a constant value due to equilibrium between the and forms. e.g. freshly prepared solution of -glucose has a specific rotation +112o. When this solution is allowed to stand the rotation falls till reach + 52.7o. The equilibrium reached is: 36% –D- glucose D = + 18.7 0 64% –D- glucose D = + 112 0 The sum is + 52.7o Terms used to describe isomerism: Glucose and Galactose are different from each other in the stereochemistry of carbon 4. They are described as “Epimers”. Glucose and Mannose are different from each other in the stereochemistry of carbon 2. They are described as “Epimers”. Epimers: CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose CHO CH OH HO CH HO CH CH OH CH2OH D-Galactose CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose CHO HO CH HO CH CH OH CH OH CH2OH D- Mannose Galactose and Mannose are different from each other in the stereochemistry of carbons 2 and 4. They are described as “Diastereoisomers””Diastereomers”. Diastereomers: CHO CH OH HO CH HO CH CH OH CH2OH CHO HO CH HO CH CH OH CH OH CH2OH D-Galactose D- Mannose Glucose and Fructose have the same molecular formula C6H12O6. They have different structures with different function groups. They are described as “Structural Isomers”. Structural isomers: CHO CH OH HO CH CH OH CH OH CH2OH CH2OH CO HO CH CH OH CH OH CH2OH D-Glucose D-Fructose Chemical Reaction Related to Colour Tests for Carbohydrates Effect of conc. acids: Treatment with conc. mineral acid (HCl or H2SO4) leads to dehydration of sugars and formation of the corresponding furfural. Pentoses O CHO Hexoses HOH2C O CHO Furfural Volatile 5-Hydroxymethyl furfural less volatile Reaction of furfural with amines resulted in Shiff’s bases with different colours used as colour tests such as: Molisch’s test: Any carbohydrate + Alcoholic -naphthol then add conc. H2SO4 on the wall of the test tube Violet ring between the two layers. Resorcinol test (for keto-hexoses): Sugar solution + few crystals of Resorcinol + Equall volume of conc. HCl and warm on water bath Rose Red Colour. Furfural test (Differentiate between Pentoses and Hexoses): Pentose + Coc. Acid and heat, expose the vapours to Aniline acetate paper Red colour Hexoses give negative result. Effect of Alkalis Strong alkalis: Polymerization. Weak alkalis: Isomerization. H C=O CH OH HO CH CH OH CH OH CH2OH CHOH C OH HO CH CH OH CH OH CH2OH D-Glucose CH2OH C=O HO CH CH OH CH OH CH2OH D-Fructose CHO HO CH HO CH CH OH CH OH CH2OH D- Mannose Oxidation Mild Oxidation: These are oxidizing agents like Bromine water that convert the CHO group to COOH to produce “onic acids”. Colour tests based on this reaction: Fehling’s reduction test: Sugar solutions + Fehling’s A (CuSO4) + Fehling’s B (NaOH, NaK tartarate rochell salt), heat on water bath Red Precipitate RCHO + Cu++ CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose RCOOH + Cu2O I2 or Fehling's COOH CH OH HO CH CH OH CH OH CH2OH Gluconic acid Barfoed’s test: Sugar solution + Barfoed’s reagent (Cu Acetate/Acetic acid), heat for 3 minutes on boiling water bath Red ppt with monosaccharides only. Acidic medium decreases the oxidation power of Cu++. Strong Oxidation These are oxidizing agents like HNO3 that convert the CHO and CH2OH group to COOH to produce “aric acids”. CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose HNO3 COOH CH OH HO CH CH OH CH OH COOH Saccharic acid Glalactaric acid (Mucic acid) test: Oxidation of galactose resulted in the formation of Galactaric acid. It is a meso compound insoluble in water and have zero optical rotation. CHO CH OH HO CH HO CH CH OH CH2OH D-Galactose HNO3 COOH CH OH HO CH HO CH CH OH COOH Galactaric acid Enzymatic oxidation Takes place in plants and resulted in the oxidation of the primary alcohol group only producing “uronic acids”. CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose CHO CH OH HO CH CH OH CH OH COOH Glucuronic acid COOH O H H OH OHOH H H H OH Reduction This resulted in the reduction of the CHO to CH2OH producing “Sugar Alcohols”. Sodium borohydride or H2/Pt are examples of reducing agents. Glucose Galactose Mannose CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose Sorbitol Dulcitol Mannitol CH2OH CH OH HO CH CH OH CH OH CH2OH Sorbitol Pentoses Examples: -D-Ribose: found in all plant and animal cells as the carbohydrate part of nucleic acids e.g. ribonucleic acid (RNA). -D-Xylose (or wood sugar): prepared from corncobs, bran, straw (or any woody material) by boiling with acids, fermenting out any glucose present with yeast, and crystallizing the D-xylose from the evaporated solution. -L-Arabinose (or pectin sugar): found in gums, pectic substances, accompanying hemicelluloses and forms the sugar part of several glycosides. CHO CHO CHO CH OH CH OH CH OH HO CH CH OH HO CH CH OH CH OH CH2OH CH2OH CH2OH L-Arabinose D-Ribose HO CH D-Xylose Hexoses 1- -D-Glucose (dextrose, grape sugar, blood sugar or cornmon sugar) Occurrence: Widely distributed in nature. Present in Grape and blood. Preparation: D-Glucose is commercially prepared from starch by: Autoclaving (at 150 0C) an aqueous starch suspension (15-20%) with dilute acid (0.03 N hydrochloric acid) for 30 minutes (complete hydrolysis). CHO CH OH HO CH CH OH CH OH CH2OH D-Glucose CH2OH O OH H H OH H H OH H OH D-Glucopyranose Uses: As source of energy either by mouth or IV injection. IV solutions to restore blood volume. Shocks following insulin administration. As osmotic diuretic. Sweetening agent for Pharmaceutical preparations, ice-cream and candy. Liquid glucose: Preparation: It is prepared by partial acid hydrolysis of starch using dilute hydrochloric acid and heating for 20 minutes at about 30 pounds pressure. Composition: It consists of a mixture of glucose, dextrin, maltose and water. Uses: Used as sweetening agent, as substitute for sucrose and as an excipient in massing pills. 2- Fructose (Levulose, Fruit Sugar) Preparation: Acid hydrolysis of Inulin. Hydrolysis of Sucrose. Sucrose Acid or Enzyme Fructose + Glucose Ca(OH) 2 Ca fructosate ppt Ca glucosate Solution CH2OH C O HO CH CH OH CH OH CH2OH D-(-)-Fructose Uses: Infant food. Diabetic food. Diet control. HOH2C O OH HO CH2OH OH OH -D-(-)-Fructofuranose Some Monosaccharide derivatives in Pharmacy and Medicine Gluconic acid and its salts: Preparation: Gluconic acid is prepared from glucose by mild oxidation using either dilute HNO3 or Br2/Na2CO3 or Electrically or by fermentation using Acetobacter aceti. Uses: Ca gluconate is used (by i.v. or orally) for treatment calcium deficiency. Ferrous gluconate, (orally or by i.v.) is used in iron deficiency. These salts are characterized by being more easily adsorbed than other Ca or Fe salts. Glucuronic Acid: Naturally present in Gums and Mucilage's. It can be prepared by Enzymatic oxidation of glucose. Uses: Treatment of certain arthritic condition as it is a component of cartilages, joint capsules and fluids, nerve sheath and tendons. Aurothioglucose: Treatment of rheumatic arthritis by IM injection. CH2OH O H H H OH OH H H S Au OH Aurothioglucose Auranofin: It is the alkyl Phosphine Gold complex with Acetylated thioglucose. Treatment of rheumatic arthritis orally. CH2OR O S H H OR Sorbitol and Mannitol: Preparation: OR H H H Au P(C2H5)3 R= COCH3 OR Auranofin Sorbitol is prepared by reduction of glucose and mannitol by reduction of mannose. Uses: Sorbitol is used as a mild laxative, in preparation of spans and tweens, sweetening agent, osmotic diuretic and in some food and cosmetics industries. Mannitol is used as an osmotic diuretic, laxative, , vasodilator (mannitol hexanitrate), antineoplastic (mannitol- (mannitol hexanitrate). nitrogen mustard) and in laboratory diagnosis of kidney function.