Carbohydrates (Part -1) - Chemistry of Natural Products, Organic Chemistry, CSIR-NET - Government Jobs PDF Download

Carbohydrates

Carbohydrates are the most abundant class of organic compounds found in living organisms. They originate as products of photosynthesis, an endothermic reductive condensation of carbon dioxide requiring light energy and the pigment chlorophyll.

n CO₂ + nH₂O + energy    C_nH_2nO_n  + n O₂

As noted here, the formulas of many carbohydrates can be written as carbon hydrates, C_n(H₂O)_n, hence their name. The carbohydrates are a major source of metabolic energy, both for plants and for animals that depend on plants for food. Aside from the sugars and starches that meet this vital nutritional role, carbohydrates also serve as a structural material (cellulose), a component of the energy transport compound ATP, recognition sites on cell surfaces, and one of three essential components of DNA and RNA.
Carbohydrates are called saccharides or, if they are relatively small, sugars. Several classifications of carbohydrates have proven useful, and are outlined in the following table.

1. Glucose

Carbohydrates have been given non-systematic names, although the suffix ose is generally used. The most common carbohydrate is glucose (C₆H₁₂O₆). Applying the terms defined above, glucose is a monosaccharide, an aldohexose (note that the function and size classifications are combined in one word) and a reducing sugar. The general structure of glucose and many other aldohexoses was established by simple chemical reactions. The following diagram illustrates the kind of evidence considered, although some of the reagents shown here are different from those used by the original scientists.

Hot hydriodic acid (HI) was often used to reductively remove oxygen functional groups from a molecule, and in the case of glucose this treatment gave hexane (in low yield). From this it was concluded that the six carbons are in an unbranched chain. The presence of an aldehyde carbonyl group was deduced from cyanohydrin formation, its reduction to the hexa-alcohol sorbitol, also called glucitol, and mild oxidation to the mono-carboxylic acid, glucuronic acid. Somewhat stronger oxidation by dilute nitric acid gave the diacid, glucaric acid, supporting the proposal of a six-carbon chain. The five oxygens remaining in glucose after the aldehyde was accounted for were thought to be in hydroxyl groups, since a penta-acetate derivative could be made. These hydroxyl groups were assigned, one each, to the last five carbon atoms, because geminal hydroxyl groups are normally unstable relative to the carbonyl compound formed by loss of water. By clicking on the above diagram, it will change to display the suggested products and the gross structure of glucose. The four middle carbon atoms in the glucose chain are centers of chirality and are colored red.

Glucose and other saccharides are extensively cleaved by periodic acid, thanks to the abundance of vicinal diol moieties in their structure. This oxidative cleavage, known as the Malaprade reaction is particularly useful for the analysis of selective O-substituted derivatives of saccharides, since ether functions do not react. The stoichiometry of aldohexose cleavage is shown in the following equation.

The Configuration of Glucose

The four chiral centers in glucose indicate there may be as many as sixteen (2⁴) stereoisomers having this constitution. These would exist as eight diastereomeric pairs of enantiomers, and the initial challenge was to determine which of the eight corresponded to glucose. This challenge was accepted and met in 1891 by the German chemist Emil Fischer. His successful negotiation of the stereochemical maze presented by the aldohexoses was a logical tour de force, and it is fitting that he received the 1902 Nobel Prize for chemistry for this accomplishment. One of the first tasks faced by Fischer was to devise a method of representing the configuration of each chiral center in an unambiguous manner. To this end, he invented a simple technique for drawing chains of chiral centers, that we now call the Fischer projection formula. Click on this link for a review. 

At the time Fischer undertook the glucose project it was not possible to establish the absolute configuration of an enantiomer. Consequently, Fischer made an arbitrary choice for (+)-glucose and established a network of related aldose configurations that he called the D-family. The mirror images of these configurations were then designated the L-family of aldoses. To illustrate using present day knowledge, Fischer projection formulas and names for the D-aldose family (three to six-carbon atoms) are shown below, with the asymmetric carbon atoms (chiral centers) colored red. The last chiral center in an aldose chain (farthest from the aldehyde group) was chosen by Fischer as the D / L designator site. If the hydroxyl group in the projection formula pointed to the right, it was defined as a member of the D-family. A left directed hydroxyl group (the mirror image) then represented the L-family. Fischer's initial assignment of the D-configuration had a 50:50 chance of being right, but all his subsequent conclusions concerning the relative configurations of various aldoses were soundly based. In 1951 x-ray fluorescence studies of (+)-tartaric acid, carried out in the Netherlands by Johannes Martin Bijvoet (pronounced "buy foot"), proved that Fischer's choice was correct.

It is important to recognize that the sign of a compound's specific rotation (an experimental number) does not correlate with its configuration (D or L). It is a simple matter to measure an optical rotation with a polarimeter. Determining an absolute configuration usually requires chemical interconversion with known compounds by stereospecific reaction paths.

Models of representative aldoses may be examined by clicking on the Fischer formulas for glyceraldehyde, erythrose, threose, ribose, arabinose, allose, altrose, glucose or mannose in the above diagram.

2. Important Reactions

Emil Fischer made use of several key reactions in the course of his carbohydrate studies. These are described here, together with the information that each delivers.

Oxidation

As noted above, sugars may be classified as reducing or non-reducing based on their reactivity with Tollens', Benedict's or Fehling's reagents. If a sugar is oxidized by these reagents it is called reducing, since the oxidant (Ag⁽⁺⁾ or Cu⁽⁺²⁾) is reduced in the reaction, as evidenced by formation of a silver mirror or precipitation of cuprous oxide. The Tollens' test is commonly used to detect aldehyde functions; and because of the facile interconversion of ketoses and aldoses under the basic conditions of this test, ketoses such as fructose also react and are classified as reducing sugars.

When the aldehyde function of an aldose is oxidized to a carboxylic acid the product is called an aldonic acid. Because of the 2º hydroxyl functions that are also present in these compounds, a mild oxidizing agent such as hypobromite must be used for this conversion (equation 1). If both ends of an aldose chain are oxidized to carboxylic acids the product is called an aldaric acid. By converting an aldose to its corresponding aldaric acid derivative, the ends of the chain become identical (this could also be accomplished by reducing the aldehyde to CH₂OH, as noted below). Such an operation will disclose any latent symmetry in the remaining molecule. Thus, ribose, xylose, allose and galactose yield achiral aldaric acids which are, of course, not optically active. The ribose oxidation is shown in equation 2 below.

Other aldose sugars may give identical chiral aldaric acid products, implying a unique configurational relationship. The examples of arabinose and lyxose shown in equation 3 above illustrate this result. Remember, a Fischer projection formula may be rotated by 180º in the plane of projection without changing its configuration.

Reduction

Sodium borohydride reduction of an aldose makes the ends of the resulting alditol chain identical, HOCH₂(CHOH)_nCH₂OH, thereby accomplishing the same configurational change produced by oxidation to an aldaric acid. Thus, allitol and galactitol from reduction of allose and galactose are achiral, and altrose and talose are reduced to the same chiral alditol. A summary of these redox reactions, and derivative nomenclature is given in the following table.

Derivatives of HOCH₂(CHOH)_nCHO

Osazone Formation

The osazone reaction was developed and used by Emil Fischer to identify aldose sugars differing in configuration only at the alpha-carbon. The upper equation shows the general form of the osazone reaction, which effects an alpha-carbon oxidation with formation of a bis-phenylhydrazone, known as an osazone. Application of the osazone reaction to D-glucose and D-mannose demonstrates that these compounds differ in configuration only at C-2.

Chain Shortening and Lengthening

These two procedures permit an aldose of a given size to be related to homologous smaller and larger aldoses. The importance of these relationships may be seen in the array of aldose structurespresented earlier, where the structural connections are given by the dashed blue lines. Thus Ruff degradation of the pentose arabinose gives the tetrose erythrose. Working in the opposite direction, a Kiliani-Fischer synthesis applied to arabinose gives a mixture of glucose and mannose. An alternative chain shortening procedure known as the Wohl degradation is essentially the reverse of the Kiliani-Fischer synthesis.

Using these reactions we can now follow Fischer's train of logic in assigning the configuration of D-glucose.

1. Ribose and arabinose (two well known pentoses) both gave erythrose on Ruff degradation. As expected, Kiliani-Fischer synthesis applied to erythrose gave a mixture of ribose and arabinose. 
2. Oxidation of erythrose gave an achiral (optically inactive) aldaric acid. This defines the configuration of erythrose.
3. Oxidation of ribose gave an achiral (optically inactive) aldaric acid. This defines the configuration of both ribose and arabinose.
4. Ruff shortening of glucose gave arabinose, and Kiliani-Fischer synthesis applied to arabinose gave a mixture of glucose and mannose.
5. Glucose and mannose are therefore epimers at C-2, a fact confirmed by the common product from their osazone reactions.
6. A pair of structures for these epimers can be written, but which is glucose and which is mannose?

In order to determine which of these epimers was glucose, Fischer made use of the inherent C₂ symmetry in the four-carbon dissymmetric core of one epimer (B). This is shown in the following diagram by a red dot where the symmetry axis passes through the projection formula. Because of this symmetry, if the aldehyde and 1º-alcohol functions at the ends of the chain are exchanged, epimer B would be unchanged; whereas A would be converted to a different compound. By clicking on the diagram, the consequences of such an exchange will be displayed. 

Fischer looked for and discovered a second aldohexose that represented the end group exchange for the epimer lacking the latent C₂ symmetry (A). This compound was L-(+)-gulose, and its exchange relationship to D-(+)-glucose was demonstrated by oxidation to a common aldaric acid product. Equations for this operation will be displayed by clicking again on the above diagram. The remaining epimer is therefore mannose.

3. Ketoses

If a monosaccharide has a carbonyl function on one of the inner atoms of the carbon chain it is classified as a ketose. Dihydroxyacetone may not be a sugar, but it is included as the ketose analog of glyceraldehyde. The carbonyl group is commonly found at C-2, as illustrated by the following examples (chiral centers are colored red). As expected, the carbonyl function of a ketose may be reduced by sodium borohydride, usually to a mixture of epimeric products. D-Fructose, the sweetest of the common natural sugars, is for example reduced to a mixture of D-glucitol (sorbitol) and D-mannitol, named after the aldohexoses from which they may also be obtained by analogous reduction. Mannitol is itself a common natural carbohydrate.
Although the ketoses are distinct isomers of the aldose monosaccharides, the chemistry of both classes is linked due to their facile interconversion in the presence of acid or base catalysts. This interconversion, and the corresponding epimerization at sites alpha to the carbonyl functions, occurs by way of an enediol tautomeric intermediate. By clicking on the diagram, an equation illustrating these isomerizations will be displayed.

Because of base-catalyzed isomerizations of this kind, the Tollens' reagent is not useful for distinguishing aldoses from ketoses or for specific oxidation of aldoses to the corresponding aldonic acids. Oxidation by HOBr is preferred for the latter conversion.

4. Anomeric Forms of Glucose

Fischer's brilliant elucidation of the configuration of glucose did not remove all uncertainty concerning its structure. Two different crystalline forms of glucose were reported in 1895. Each of these gave all the characteristic reactions of glucose, and when dissolved in water equilibrated to the same mixture. This equilibration takes place over a period of many minutes, and the change in optical activity that occurs is called mutarotation. These facts are summarized in the diagram below.

When glucose was converted to its pentamethyl ether (reaction with excess CH₃I & AgOH), two different isomers were isolated, and neither exhibited the expected aldehyde reactions. Acid-catalyzed hydrolysis of the pentamethyl ether derivatives, however, gave a tetramethyl derivative that was oxidized by Tollen's reagent and reduced by sodium borohydride, as expected for an aldehyde. These reactions will be displayed above by clicking on the diagram.

The search for scientific truth often proceeds in stages, and the structural elucidation of glucose serves as a good example. It should be clear from the new evidence presented above, that the open chain pentahydroxyhexanal structure drawn above must be modified. Somehow a new stereogenic center must be created, and the aldehyde must be deactivated in the pentamethyl derivative. A simple solution to this dilemma is achieved by converting the open aldehyde structure for glucose into a cyclic hemiacetal, called a glucopyranose, as shown in the following diagram. The linear aldehyde is tipped on its side, and rotation about the C4-C5 bond brings the C5-hydroxyl function close to the aldehyde carbon. For ease of viewing, the six-membered hemiacetal structure is drawn as a flat hexagon, but it actually assumes a chair conformation. The hemiacetal carbon atom (C-1) becomes a new stereogenic center, commonly referred to as the anomeric carbon, and the α and β-isomers are called anomers.

We can now consider how this modification of the glucose structure accounts for the puzzling facts noted above. First, we know that hemiacetals are in equilibrium with their carbonyl and alcohol components when in solution. Consequently, fresh solutions of either alpha or beta-glucose crystals in water should establish an equilibrium mixture of both anomers, plus the open chain chain form. This will be shown above by clicking on the diagram. Note that despite the very low concentration of the open chain aldehyde in this mixture, typical chemical reactions of aldehydes take place rapidly.
Second, a pentamethyl ether derivative of the pyranose structure converts the hemiacetal function to an acetal. Acetals are stable to base, so this product should not react with Tollen's reagent or be reduced by sodium borohydride. Acid hydrolysis of acetals regenerates the carbonyl and alcohol components, and in the case of the glucose derivative this will be a tetramethyl ether of the pyranose hemiacetal. This compound will, of course, undergo typical aldehyde reactions. By clicking on the diagram a second time this relationship will be displayed above.