UNIVERSITY OF KWAZULU-NATAL COLLEGE OF AGRICULTURE, ENGINEERING AND SCIENCE SCHOOL OF LIFE SCIENCES BIOC201W1 INTRODUCTION TO BIOMOLECULES . on OH nace ? ' '° ° H-¢-0H Se ° + HO-G-H *, H=C-OH * H-=C-OH * HO 2 / a Pyran H IH Glucopyranose (Ring form of glucose) ‘The six-membered pyranose ring is not planar. Rather the preferred conformation is a chair form. HSimilarly, a ketone can react with an alcohol to form a hemiketal. The C2 ketogroup in the open chain form of fructose reacts with the C-5 hydroxyl group to form an intramolecular hemiketal. This S-membered sugar ring is called furanose because of its similarly to furan. —< a + HORS \ Alcohol ee ‘OH Hemiketal 4'CH,OH | . h 6 2 cH,OH 2c——o HOH OH CH,OH = HOHC - as ek H ch HOW OH 4 =¢ —OH | OH OH \ r) H,¢—oH o Fructose open chain Fructofuranose A ring form of fructose 9 Y u™ “c—H | H—*c —oH 1 1 —H H —'c —On 1 H —*c —OoH I *CH,OH GLUCOSE OPEN CHAIN GLUCOFURANOSE)b) STEREOISOMERISM : Compounds that have the same structural formula but differ in spacial configuration are known as stereoisomers. The formation of stereoisomers is allowed by the presence of asymmetric carbon atoms. a) ASYMMETRY: A carbon atom to which four different atoms or groups of atoms are attached is said to be asymmetric. All monosaccharides except dihydroxyacetone contain 1 or more asymmetric (chiral) carbon atoms. For example glyceraldehyde possess singly asymmetric carbon and there are two stereoisomers : D and L : Since D and L Glyceraldehyde are mirror images they are called enantiomeric pairs. ° oO i I fame C—H HC — OH HO—C— H H,COH H,COH d-glyceraldehyde glyceraldehyde The designation of all other carbonhydrates as D or L isomers is determined by its spacial relationship to the parent molecule of glyceraldehyde. By analogy the orientation of the OH group of the carbon next to the last determines whether the sugar is D or L. When the OH group is on the right, the sugar is D, when it is on the left, itis L (D = dextro = right; L = levo = left). For example glucose can be D or L form, EPIMERS, isomers differing as a result of variation in configuration of the H and OH on a single carbon atom (2 or 3 or 4) of the hexose molecules are known as epimers. Biologically the most important epimers of glucose are : galactose and mannose, formed by epimerization at carbon2 and 4 respectively CH,OH CH,OH CH,OH HO 0. 0, OH OH >) OH OH OH HO OH HO OH OH OH D-Galactose D-Glucose D-Mannose On the other hand, there is no epimeric relationship between D-Mannose and D- Galactose because they have different spacial configuration at more than one carbon atoms (C-2 and C-4). When two stereo isomers are neither enantiomers nor epimers they are called diastereomers.¢) ANOMERS : An additional asymmetric center is created when glucose cyclases. C-l, the first carbon atom in the open-chain form becomes an asymmetric centre in the ring. Two ring structures canbe formed : a-D-Glucose and B-D-Glucose. The designation alpha means that the hydroxyl group, attached to C-1 is below the plane of the ring; beta means that it is above theplane of the ring. C-1 is called anomeric carbon and o and B forms are anomers. The number of asymmetric atoms arises to 5, and 26 = 32. OH,0R CH,OH CHOR CH,OH ¢ 2 4 oH HO no | 2.08 208 oH x HO HO re iH a-D-Galactose £-D-Galactose a-D-Glucose B-D-Glucose C) OPTIMALISOMERISM AND OPTICAL ACTIVITY OF CARBOHYDRATES : The presence of asymmetric carbona toms also confers optical activity on the compounds. D and L glyceraldehydes looklike mirror images. They constitute an enantiomeric pair. These compounds are different like the right and left hand. Almost all properties of the two members of the pair are identical : they have the same meltingpoints, boiling point the same solubility and so on, but theyd iffer in their optical activity : D-Glyceraldehyde rotates a plane of the polarized light in a clockwise direction and its therefore said to be dextrarotatory (dextro, +). Its mirror image L-isomer rotates the plane of the polarized light in the opposite or counterclockwise direction and is said to be levorotatory (levo, -). ‘Other compounds with more than 1 asymmetric C may be designated D(+), D(-), L(+) or L(-) indicating structural relationship to D or L glyceraldehyde but not necessarily exhibiting the same rotation. For example the natural fructose is D(-) isomer, and the natural glucose is De). MUTAROTATION The anomers c-D(+) glucose and B-D(+)-glucose are optical isomers but not mirror images. They are also diastereomers and have different physical properties and can be separated by crystalization. If D-glucose is dissolved in ethylalcohol and is allowed to crystallize the a-form of the sugar is obtained. If D-glucose is crystallized from acetic acid, the B-anomer is obtained. The two forms have diffeent physical properties. melting point specific rotation t-form 146°C +112" B-form 150°C + 19° Both a- and B-forms are stable in the crystalline form. When either of isomers is dissolved in water, the optical rotation of each gradually changes with time and approaches a final equilibrium value of +53°. This is known as mutarotation. This value is not the average 65,5 of the rotation of a and 8B-glucose. Probably one of the isomers predominates. We can calculate the percentages of a and glucoses in the equilibrium mixture: Let us admit that in 100ml of the equilibrium mixture we have Xml of B-isomer and yl of a- isomer: Therefore : x+y=100 100 OR x+y=lorx x.19° + y.112° = 53° ‘The percentage of the B-isomer multiplied by its optimal rotation plus the percentage of the a-isomer multiplied by its optical rotation must equal the optical rotation of the final solution. Solving for x and y in two equations, we will obtain : y = 0.366 = 37% x= 1-y= 63% This gradual change of optical rotation, which continues until equilibrium is established, is known ‘as mutarotation : CHZ0H Sg |ee cH0H OH, é oH — 4) ———— iH x a-D-Glucose OPEN-CHAIN FORM —_-D-GLUCOSE This phenomenon is common to all monosaccharides (and to some disaccharides) that can exist in cand B-cyclic structures. There is evidence that all monosaccharides that undergo mutarotation pass through the open chain form. It is for this reason that the monosaccharides are said to have a potential carbonyl group.DISACCHARIDES Naturally occurring carbohydrates usually contain more than one monosaccharide unit, Monosaccharides have a tendency to react with hydroxy compounds to form stable acetals called glycosides. These glycosides may be named according to the sugers from which they are derived, eg. glucoside from glucose, galactoside from galactose etc. The linkage (carbon-oxygen-carbon) joining the two components of the acetal is called the glycosidic linkage. The disaccharides all have the same molecular formula, C,,H,,0,, and hence they are structural isomers of one another. The component sugars of disaccharides may be identical or different: eg. maltose © = —Deglucose + = —-Deglucose sucrose = Deglucose + — Defructose lactose = Deglucose + — Degalactose cellobiose = D-glucose + — Deglucose Maltose Maltose is formed by the action of the enzyme diastate (in plants)or ptyalin (in animals) on starch. Further hydrolysis of maltose, catalysed by the enzyme maltase, yields D-glucose units only. The two a-D-glucopyranose components are joined head-to-tail through C, of one glucose molecule to C, of the second glucose molecule. The linkage is an a-1, 4-glucosidic linkage. Maltose is a reducing sugar. 4 OH 4 OH x-Daunnse Pe-cumcose xr mise rene o1ASTAT( PLAS) Hromorsts HO: HO conoensarion 7 pvat tn CR) H,OH H scene OH enacarc ce 6D yk Ay a 0 OH o ty aaa i p -macrose ML TOSE uraporaricn ALDEHVOE EXTEREDIATE 10Lactose Lactose is one of the few carbohydrates associated exclusively with the animal kingdom. It is found in milk (human : 5-8%; cow : 4-6%). Lactose may be hydrolysed by dilute mineral acid or by enzyme lactase to yield equal concentrations of D-glucose and D-galactose. The monosaccharide units of lactose are joined by a B-1,4-galactosidic linkage. The equilibrium mixture of lactose has a specific rotation of +55°. The a-form of lactose is used in the production of infant food and also penicillin, 5CH,OH H OH H IH P-o-cauacropvnanose oni-0-sLucoPrRenose creme icse SrsosfF0 eooesiion HOH sCH,OH aceTaL emacera _ HO, 0 4 AH i 2 0. oR No ¢\OH ; Ne Af uveestore H OH tine OH i p -tcrose of Lactose USED TN pRooUCTION OF INFANT FOOD AN PENICILLIN MUTAROTATION /equicrentum mixruRe HAS SPECIFIC. ROTATION OF +559 5CH,OH 6CH,OH HO A, a 0. A” gotentiat OH Sy earbon ern H OH 4 ALDEHYDE INTERMEDIATE MWCellubiose Cetlubiose is a stereoisomer of maltose. It isa product of hydrolysis of cellulose (a polysaccharide) by fungi. It also contains 2 glucose units, which are joined by a-1,4-glucosidic bond. Like maltose cellubiose is a reducing sugar and undergoes mutarotation. 7 -0-s.000se on rota -2-61u008e B-D- 2 exrcosrore {0H LINK CH,OH a 1H 8-D-Froctose only Sucrose 13POLYSACCHARIDES The overwhelming bulk of all carbohydrates in nature are the polysaccharides.. They serve a structural function or play an important role as a stored form of energy or fuel. The general formula is (CcH,g0,),. The polysaccharides in the nature are : starch, glycogen, cellulose, chiten, dextrins, glycoproteins. All polysaccharides can be hydrolyzed with acid or enzymes to yield monosaccharides. Those polysaccharides that on hydrolysis yield only a single type of monosaccharides molecule are termed Homopolysaccharides. Heteropolysaccharides on hydrolysis yield a mixture of constituent monosaccharides and derived products. | | HOMOPOLYSACCHARIDES HETROPOLYSACCHARIDES Homopolysaccharides 1. Storage Fuel homopolysaccharides a) Starch is the most important food source of carbohydrate and is found in cereals, potatoes, legumes, and other vegetables and it is an end product of photosynthesis. It is the source of fuel or energy in plants growth and respiration. The two chief constituents are : Amylose (98%); which is non-branching in structure, and Amylopection (2%) which consists of highly branched chains The amylose component consists of D-glucose units linked in linear fashion by @(1-4) linkages; it has a non-reducing end and a reducing end. Its molecular weight can vary from few thousand to 150 000. Amylose gives a characteristic blue color with iodine due to the ability of the halide to occupy a position in the interior of a helical coil of glucose units that is formed when amylose is suspended in water. Amylopectin is a branched polysaccharide; in this molecule, shorter chains (about 24-30 units) of glucose molecules linked by a(1-4) linkage are also joined to other chains by «(1-6) linkings. Amylopectin produces a purple to red colour with iodine. Since the helix formation is interrupted by branch points («1->6) hence yielding less intensely coloured complexes with iodine. b) Glycogen is the storage polysaccharide of animals and is the source of fuel and energy in man and animals for growth, metabolism and respiration. It is often called animal starch. It is similar in structure to amylopectin in that it is branched homopolysaccharide composed of glucose units. But it is more highly branched than amylopectin, having branch points about every 8 to 10 glucose units. It is non-reducing sugar and gives a red color with iodine. Since the greater number of branch points allow a less dense helix to form. Less helix means less stain taken up by the molecule into the helical cods. 14CHZOH CH,OH OH ~ . E i iH oH OH oe Reou v7? HY es) CING H on Ih END oH 4 REDUCING END Hu by ctrOH Vis fo 4 ic) OH HY H @I>b) © brawoH 7 pr CH30H4 oh, ies KT at oH ie }. on a) 4 oH H on Ho: em, oO ns0H é wo on ? (a1aflntedrglucase units Most stuble Confer mation, with adjacent rigid chairs chain 15 curved: These links couse these polymers to coiled assume a bight! re. tSb) Structural homopolysaccharides Cellulose is the chief constituent of the framework of plants. Cellulose is a linear homopolymer of D-glucose units linked by B(1-4) glycosidic bonds. Instead of forming a coiled helix, cellulose forms a structure of parallel chains that are cross linked by hydrogen bonding. Due to the lack of helix formation, cellulose gives no colour when reacted with iodine. Hon CHO ree EEE ER | \ Several chains lying side by side can form a stabilizing network of inter and intra chain hydrogen bonds resulting in the straight stable supramolecular fibres of great tensile strength. This tensile strength of cellulose has made it a useful substance to civilization, Many manufactured products, including paper, cardboard, insulating tiles and other packaging and building materials are derived from cellulose. rt vk [>4) linked Ae unibs In contranst to starch, the (1-4) linkages of cellulose are resistant to hydrolysis and are not hydrolyzed by the amylases, thus man and most animals cannot utilize the energy present in cellulose. Ruminants are an important exception, however, since the bacteria that reside within the rumen secrete cellulase, a B-glucosidase, that catalyzes the hydrolysis of cellulose. Termites can also degrade cellulose because their digestive tract conainsa parasite (unicellular cilliate) that secretes cellulase. Chiten is a linear homopolysaccharide composed of N-acetyl-D glucosamine residues in a B linkage. The only chemical difference from cellulose is the replacement of a OH group at C2 with an acetylated amino acid. Chiten forms extended fibres similar to those of cellulose and like cellulose is indigestable by vertebrate animals. Chitin is the principle component of ee hard exoskeletons of arthropods, such as insects, crabs, beetles, lobsters, crayfish and so CHOH CHO a Np c =0 c=0 I | CHs CH, 16Heteropolysaccharides Heteropolysaccharides are 1) glycoproteins, 2) glycolipids and 3) peptidoglycan, 4) glycosamino- glycogen and proteoglycans. 1 ycosaminoglycans and Proteoglycans - components of extracellular matrix The extracellular space in animal tissues is filled with a gel-like material, the extracellular matrix, also called ground substance which holds the cells of a tissue together and provides a porous pathway for the diffusion of nutrients and oxygen to the individual cells. The extracellular matrix is composed of an interlockingnetwork of heteropolysaccharides and fibrous proteins. These heteropolysaccharides called glycosaminoglycans are a family of linear polymers composed of repeating disaccharide units. The glycosaminoglycan hyaluronic acid or hyaluronate at physiological pH of the extracellular matrix of animal tissues contains alternating units of D-glucuronic acid and N-acetyl glucosamine. Hyaluronates have molecular weights greater than 1 million. They form clear highly viscous solutions which serve as lubricants in the synoviral fluid of joints, found in the umbilical cord and give the vitrous humor of the eye its jellylike consistency. Hyaluronate is also a central component of the extracellular matrix of cartliage and tendons to which it contributes tensile strength and elasticity. Glycosaminoglycan Repeating Disaccharide No. of Disaccharide per chain HYALURONATE ~ 50 000 pisos H 0 coo | 4 : a Ho oy HAW HH HP oH H MH Hpie3 C=O HOH CH; Gle uA S IeNAc Hyaluronidase, an enzyme secreted by some dangerous bacteria make tissue more susceptible to bacterial invasion and infection. A similar enzyme in sperm, hydrolyses the outer glycosaminoglycan coat around the ovum of many organisms allowing sperm penetration. 7Proteoglveans are composed of a very long. strand of hyaluronate to which numerous molecules of core protein are bound at regular intervals. A typical proteoglycan being found in human cartilage. Peptidogivean : The rigid component of bacterial cell walls is a heteropolymer of alternating B(1—4) linked N-acetylmuramic acid (Mur Nac) units, ‘Many such linear polymers lie side by side in the cell wall, crosslinked by short peptides, the exact structure of which depends on the bacterial species. ‘This cross-linked peptidoglycan is degraded by an enzyme lyzozyme, which hydrolyses the glycosidic bond between these 2 sugar units ultimately killing the bacterial cells. Lysozyme is present in tears, presumably a defense against bacterial infection of the eye. Clanage by tory cH 1 R= ba fn oO tae Nea Uae ! 7 cE aetite : piala oH chon weir WI 4 ee Sys iA ore) on fe fro bey bus Glewae martiAe Glycoproteins and Glycolipids Most of the oligo- and polysaccharides in animals and plants are linked covalently to protein or lipid molecules known as glycoproteins or glycolipids. 18CARBO) E CHEMISTRY 1. Oxidation of sugars Carbohydrates may be classified as either - reducing or non-reducing sugars. Although all polysaccharides (starch, glycogen, cellulose, pectin, chitin) are regarded as non-reducing note that each bears one reducing end. The disaccharide sucrose is a non-reducing sugar. Reducing sugars comprise all the other saccharides which are able to function as reducing agents because free ot potentially free aldehydes and ketone groups are present in the molecule. The reducing properties of sugars are usually observed by their ability to reduce the metal ions Cu”, Ag’, Bi in alkaline solution. These properties are utilized in both qualitative and quantitative analyses of sugars. When we treat sugars with dilute alkaline solution aldoses and ketoses isomerise:- HO=0 HO- G—H HO—CH, i H— C—OH C— OH c=0 i on- | ou ! Ho—c—H 24. no-c—H 4) no—c—H 1 I i [ H— C—on H—¢ —oH H— C—oH i H— ¢—oH H—¢—oH H—C—on CH,OH CH,OH du,0n D-Glucose Trans-Endiol D-Fruetose o=c—H HO-E—H gy” uo—b—u _ HO-¢ Ho—¢ ou { —C —H — Roc 1 ame oe H—¢—on H#—¢ —ox H— © —OoH H—C—oi | 1 CH,OH CH,OH D-Mannose Cis-Endiol D-Glucose, D-Mannose and D-Fructose form the same endiol in dilute alkali. If any one of these sugars is allowed to stand in dilute alkali for some length of time, and the solution is subsequently acidified, a mixture of all three sugars will be found. All monosaccharides, aldohexoses and aldopentoses, ketohexoses andketopentose monosaccharide sugars containing free reducible groups (i.e. CHO or C== 0) and adjacent free hydroxyl group tautomerise (enolisation) in dilute aqueous alkaline solution forming their own typical endiol intermediates or isomers. These endiol forms of the sugars are highly reactive intermediates which are readily oxidized. They easily reduce oxidising ions (Cu, Ag’, Bi* and Fe (CN). We use these properties to detect the reducing sugars in biological fluids. For example Benedicts test is sufficiently sensitive to detect reducing sugars in urine, These endiol sugars then reduce the Cu’ (cupric ion) of Benedicts reagent to Cu (cuprous ion) which is less soluble and Cu,O (cuprous oxide) preapitates out of the solution asa yellow or red solid. The reducing sugar is in tum oxidised to the corresponding carboxylic acid. 19SUGAR + ALKALI. Mixture of Er@diols eg. Glucose == 2cu* Cupric ion as a complex of citrate or tartarate 2cu* Sugar Acids Cuprous ion OH alkali Cu, (OH), yetLow Venn HO cu,” H.C ——OH RED CUPROUS OXIDE Pounce Acid Disaccharides such as maltose, lactose and sucrose consist of monosaccharides joined by an 0- glycosidic bond, formed when the hydroxyl group on one sugar reacts with the anomeric carbon on the other. When an anomeric carbon becomes involved in a glycosidic bond it cannot be oxidised and the sugar monosaccharide containing this ‘engaged’ anomeric carbon no longer acts as a reducing sugar. In the case of maltose the anomeric carbon of one glucose molecule is involved in alycosidic bond formation with OH group on C4 of the second glucose molecule. Hence, the anomeric carbon (C1) of the 2" glucose molecule residue is available for oxidation and thus maltose isa reducing disaccharide. (Refer back to disaccharide section). suon sexu gion spon YE SOK we ee Kon ou we Kp ‘The disaccharide lactose has the anomeric C1 of galactose in glycosidic bond formation with OH group of C4 of glucose, the second monosaccharide. However, the anomeric C1 ofthe glucose residue is available for oxidation and thus lactose is a reducing disaccharide. Sucrose is adisaccharide of glucose and fructose. Sucrose containsno free anomeric carbon atoms. The anomeric carbons of both monosaccharide units are involved in the glycosidic bond. Sucroseis therefore not a reducing sugar,and has no reducing end, (Refer back to disaccharide section). 7 20H HodH, 0. “Rn nos . i on ou ; 3 Sucrose (O-a-b-glucopyranosyl-{1—+2)-8-p-fructofuranoside) oes ‘Gisai=2}¢ra i L0H m socee 202. Treatment of sugars with mineral acids Monosaccharides are generally stable in dilute mineral acids even on heating. But when aldohexoses/aldopentoses and ketose sugars are heated with strong concentrated mineral acids such as sulphuric acid, all carbohydrates are dehydrated forming appreciable amounts of furfural or furfural derivatives. The latter condense with a-naphthol and other phenolic compounds to give highly coloured pink to violet complexes. This reaction is called the Molish Test and is used for the detection of monosaccharides but also for the detection of disaccharides and even polysaccharides: sulphuric acid first hydrolyses the glycosidic bonds to give monosaccharides which are then dehydrated to furfural and hydroxy furfural. Hence the test is generally slower to react with polysaccharides. ° bat fe com. te—cu I HSO, 9 Gi ha ip © cot ay ' f 8—c5H] / | Tete How.c Bo , © gypaoxyeraye I FURFURAL ‘CHLOH (CHAO) ALDOHEXOSE S-ctueose 5 (CH.00 ° i 4 cone secu +H,SO, J l BB. A en 5 . fon 1 gy} $ 1 : agi : FURFURAL (co) aporenTose haa Dexvuose (CHO) seca He —cx L r a@-NAPHTHOL It ? < & Ny > ~c woud b/ gad W~4Q? wemernemn fo ay {O) FURFURAL B a VARIOUS COLOURED CONDENSATION PRODUCTS PINK —> VIOLET 213. Cyanohydrin Synthesis or Elongation The cyanohydrin synthesis of Killian Fisher is a process by which the chain length of an aldose is increased by one carbon atom and 2 new aldoses can be formed. Aldehydes (aldoses) react with hydrogen cyanide (HCN) to give cyanohydrins. Cyanohydrin formation creates a new as symmetric centre at the original C,. Subsequent hydrolysis yields carboxylic acids (aldonic acids) containing | extracarbon than the starting sugar. Reduction (Na/Hg) yields a mixture of 2 aldose sugars containing 1 more C atom than the original sugar. H—C,—=o R OR Aldose sugar 2 Cyanohydrins cal NH, 2H,0 (Hydrolysis) COOH cOooH | 1 H— ——0H HO—o,—H t R R Na/Hg Reduction ° ° 1 i fH C—H H H— C——OH HOC —H R R isomers Aldoses containing 1 more C atom than original sugar. For example, D Glyceraldehyde subjected to the above reactions would yield D-Erythrose and D- Threose. Similarly D-Erythrose and D-Threose subjected to cyanohydrin synthesis would result in four aldopentoses-D-ribose Darabinose D-xylose an D-lyxose (Refer back to page 4 for structures). 220 il C—H HEN oR H—C—oH ——> H— c—on CH,OH CH,OH D Glyceraldenyde 3c) Ot aac Cc==N H a ——0H NITRILE Ua apace GROUP CH,OH CYANOHYDRINS fo—t —H H——C—0H L CH,OH 2H,0 Hydrolysis NH, CARBOXYLIC ACIDS 0 == C—OH HO —t —H 4H#— ¢ —oH CH,OH S-LACTONES o—=c —a | HO— C—H | ° H——€—oH | cH, 2H+ al REDUCTION o—c H— ¢—on I 0 H—c—oH dh, o—=c—H u— bon H#— ¢-—on Gin D ERYTHROSE (4C) 23 (+1) ALDOSES Oo=c—H Ho —¢— 4—¢—on bx,0H D-THREOSE 4(C)4. Oxime Formation or Chain Shortening Hydroxylamine (NH,OH) reacts with both aldoses and ketose to form oximes. If the sugar oxime is treated with acetic anhydride (Ac,O), dehydration occurs converting the sugar oxime to a lower cyanonhydrin, The (Wohl reaction) or treatment with AgNO, and ammonia NH; result in an aldose sugar containing one carbon atom less than the original aldose sugar. sae + H—'c==NOH Hae) pe R Aldose Sugar i H——*C ——OH i R Oxime of Aldose Sugar Hio ——_ Acetylation with Ac,O *c 1 H— *c — oH R Cyanohydrin of Aidose Sugar A AQNO, + NH, ‘ HEN H—"e: | R 5. Aldol Condensation and Aldol Cleavage These are reactions which occur frequently in carbohydrate biochemistry. This reaction depends on the activity of a-hydrogens on the carbon atom adjacent to the carboxyl group and the ability of the anion formed by removal of hydrogen (proton) to be stabilized by resonance. The enolate ion is then capable to acting as a nucleophile and can attack the aldehyde group of a second sugar. CH,OH Ht HOH CH,OH I i au | e c—=o —— c=o0 — {orrd T CH,OH H— c¢—oH n—t—on 3c Ketose Sugar Enolate ion Resonance isomer 0 ay I c—H 4—¢—on H— ¢—on bu,0H buon 3¢ Aldehyde cH,OH c=o es H— C——oH 4 H— € — on fl CH,OH Hexose Suigar6. Periodate Oxidation There are three analytical methods available for studying the periodate oxidation of polyhydroxy compounds. These are:- a) iodometric b) acidimetric c) _spectrophotometric In the experiments you will deal with the iodometric technique. Compounds containing hydroxyl groups on adjacent carbon atoms are quantitatively oxidized at room temperature by an excess of periodic acid or its salts. H10, (Periodic acid) is very useful in carbohydrate analysis. This reagent will cleave C-C bonds if both carbons have hydroxyl group or if one carbon having a hydroxyl group is adjacent to another carbon with an amino group or keto or aldo oxygen. Every cleavage results in an oxidation. The carbon participating in the cleavage reaction are oxidised to the next level (e.g. alcohol —. aldehyde AND aldehyde to carboxylic acid). —¢—on cHO ‘COOH 1 | —¢—0oH as aoe rae 8 ALCOHOL GROUP ALDEHYDE GROUP CARBOXYLIC ACID CH, OH CH, OH a HY CH, OH 2 oH 0 10, 6 —> 5 ie 104 H-C-OH i 0 Formic acid oH g | Formic acié TOTAL:- GLUCOSE + 5 10; ——————> 5 FORMIC ACID + FORMALDEHYDE + 5 I0; + 25If however the hemiacetal C is involved in a glycosidic linkage or is methylated, the sites of possible cleavage are reduced due to the inability to open-out the ring structure into a straight chain. CH, OH © CH,OH 0 Maltose 0H CH, OH 5 CLEAVAGE SITES 0 Q 0 kectase) 1H, 0H 0, HOH.G q CH20H 5 3 CLEAVAGE SITES Sucrose 26