[BENZOIN CONDENSATION] A/S Projects

Multistep Synthesis Quinoxaline

of

Benzil Benzoin

Use of the condensation

This next task involves the multistep synthesis of a heterocycle from benzaldehyde. The name heterocycle infers a cyclic ring that contains an atom other than carbon, in this example, we will be synthesising a quinine which contains two nitrogen atoms. In order to obtain the heterocycle we will need to produce a 1,2-diketone which is a result of oxidising benzyl, itself being produced from the well know Benzoin condensation. The Benzoin condensation is catalysed using the vitamin thiamine and is a classic example of nucleophilic catalysis. The mechanism of the reaction is outlined below. Vitamin B1, thiamine, as its pyrophosphate derivative (shown below), is a coenzyme universally present in all living systems. It was originally discovered as an essential nutrient required to prevent the human disease beriberi, which affects the peripheral nervous system. Symptoms include pain and paralysis of the extremities, emaciation, or swelling of the body. The disease is still common in the Far East.

In biochemical terminology, thiamine functions as a coenzyme, a biological molecule that assists in enzymatic reactions. In most cases, coenzymes are directly involved in the biochemical reaction that the enzyme catalyzes since they usually bind the substrate (reactant) for the reaction. Without the coenzyme, no reaction will take place. The enzyme, which is the biological catalyst, binds the substrate, controlling stereochemistry, energetics, and entropic factors. As indicated above, the vitamin (derived from VITal AMINe) thiamine is required for many enzymatic reactions. In the current experiment, we will use thiamine to catalyze the reaction of benzaldehyde into benzoin. Most biochemical processes are no more than organic chemical reactions carried out under special conditions. Like most reactions in organic chemistry, many biochemical reactions can now be explained using familiar reaction

[BENZOIN CONDENSATION] A/S Projects mechanisms. To enhance reactivity, and to be stereoselective, enzymes are used to bind the substrate in a manner that allows only a single reaction, with stereoselectivity to occur. In addition, enzymatic reactions can be carried out in mild conditions and at moderate pH values. Reactions which involve hydrophobic (lipid loving or water hating) conditions that might not otherwise be possible in an aqueous, biological environment.

[BENZOIN CONDENSATION] A/S Projects

Safety

Wear Goggles 3,4-toluene diamine

Ethanol

NaOH, HNO3

You will need to do some after school slots for this procedure.

Procedure 1. Benzoin Condensation In this, the first step of our multistep reaction sequence, benzaldehyde will be condensed, using the thiamine coenzyme as catalyst, to produce benzoin. The same reaction can be performed using the cyanide ion. The reaction mechanism for thiamine catalysis is described above. (It is imperative that the benzaldehyde be from a newly opened bottle because of the ease of oxidation, producing benzoic acid, which will interfer with the reaction. The thiamine hydrochloride must also be from a newly opened bottle, although it is not quite as critical as for the benzaldehyde.) The overall reaction for conversion of benzaldehyde into benzoin is:

Method 1. Place 1.3 g of thiamine hydrochloride in a 50 cm3 round bottomed flask, dissolve the solid in 4 cm3 of water and add 15 cm3 of ethanol and cool the solution in an ice bath.

[BENZOIN CONDENSATION] A/S Projects 2. Add 2.5 cm3 of 3.0 moldm-3 sodium hydroxide dropwise with swirling. Dont allow the reaction to reach above 20oC. You might want to wear gloves when doing this stage. Add 7.5 cm3 of benzaldehyde to the yellow solution. Leave the reaction with a rubber seal in the top at RT until next week. Cool the reaction in an ice bath and scratch the side of the reaction vessel to induce crystallisation. Filter off your solid using suction filtration and wash the solid with 1:1 water/ethanol to remove the yellow colour. Look after your filter funnel as you will need to use this again to collect your pure sample. Recrystallise the solid from hot ethanol, and filter off the purified product. Collect your sample in a labelled tube, the best sample will be sent off for analysis, and place the sample in a drying oven for the next stage.

3. 4. 5.

6. 7.

Procedure 2. Nitric Oxide Oxidation to Benzil Starting with the -hydroxyketone benzoin (prepared in Part 1), you will prepare an oxidation product, benzil, which is an -diketone.

This oxidation can easily be done with a variety of mild oxidizing agents, including Fehling's solution (an alkaline cupric tartrate complex) or copper(II) sulfate in pyridine. In addition, benzoin could be oxidized by sodium dichromate, but the yield of benzil is lower because some of the benzoin is converted back into benzaldehyde following cleavage of the bond between the two oxidized carbon atoms, which is activated by the phenyl rings, producing benzoic acid as the final product. In this experiment, due to ease of use and consistent results, we will use nitric acid as the oxidizing agent. Method 1. Heat 4 g of benzoin and 14 cm3 of conc. Nitric acid in a HOT water bath. This reaction should be done in a fume hood! This procedure should take no more than 15 minutes.

[BENZOIN CONDENSATION] A/S Projects Becareful here as hot boiling acid is particularly hazardous. Ensure the reaction vessel is safely clamped in the boiling water bath. WEAR GLOVES. 2. 3. 4. Allow the reaction to cool and add 75 cm 3 of water to the reaction vessel. Swirl the reaction as the coagulant forms. Collect the yellow solid via suction filtration and wash the solid with water. To dry the solid you should also press down on the cake with a second piece of filter paper, using a boiling tube as a weight, to ensure all the water is removed. Take the solid and recrystallise it from ethanol. Dissolve the solid in 10 cm3 of ethanol and heat then set aside to crystallise. Collect your sample via suction filtration, dry, and store in a labelled sample tube (you are going to use this product in 2 separate steps).

5. 6.

Part 3. Preparation of Benzil Quinoxaline The final step which shows Benzil an an 1,2-diketone is a condensation reaction with a diamine leading to the formation of a heterocycle, in this case a quinone.
NH2 NH2
+

Heat
N O N

Diamine

Diketone

Method 1. Commercial 3,4-toluene diamine is usually badly discoloured and needs to purified. This is easily carried out by placing 0.5 g of diamine in a test tube. Evacuate the tube using a water aspirator.

[BENZOIN CONDENSATION] A/S Projects 2. Clamp the tube in a horizontal position and heat the bottom of the tube to sublime the solid. Once all the sample has sublimed allow the tube to cool and collect the white residue from the top of the tube. Weigh out 0.4g of benzil and 0.2 g of your purified diamine into a boiling tube. Place the boiling tube in a boiling water bath for 10 minutes. You should start to see the formation of a tan solid. Let the reaction cool and then dissolve the solid in around 5 cm3 of hot methanol and let the solution stand. Crystallisation should start. Filter the crystals (which should be colourless needles) and store in a labelled jar.

3. 4. 5. 6.

Part 4: Synthesis of Benzilic Acid

In this experiment, benzilic acid will be prepared by causing a rearrangement of the 1,2diketone benzil. The rearrangement of benzil proceeds as follows:

The driving force for the reaction is provided by the formation of a stable carboxylate salt (potassium benzilate). Once this salt is produced, acidification yields benzilic acid. The reaction can generally be used to convert aromatic diketones into -hydroxyacids. Other compounds, however, also will undergo benzilic acid type of rearrangement. Method 1. 2. Add 3.0 g of benzil and 9.0 cm3 of 95% ethanol to a 50 cm3 flask and attach a reflux condenser. Heat the mixture until the benzil is dissolved. Add dropwise 7.5 cm3 of an aqueous potassium hydroxide solution downward through the condenser into the flask. Gently reflux the mixture for 15 minutes with swirling. After the mixture has dissolved and has been heated for a few minutes, the mixture will turn blue-black in colour. As the reaction proceeds, the reaction product will turn brown and the solid may, or may not, be completely dissolved. At the end of the reaction, remove the flask and let it cool.

[BENZOIN CONDENSATION] A/S Projects 3. When the apparatus is cool enough to handle, remove the condenser and transfer the contents, including any solids, into a 150 cm3 beaker. Allow the mixture to cool to room temperature. When the mixture is cooled, continue the cooling in an ice-water bath for an additional 15 minutes, when crystallization should be complete. Crystallization is complete when it appears that virtually the entire mixture is solidified. If this does not occur in 15 minutes, allow the mixture to set overnight, or until complete (it is possible to store in an ice bucket or in a refrigerator, if necessary). Collect the crystals using vacuum filtration and wash the crystals thoroughly with three 15 cm3 portions of ice-cold 95% ethanol. The solvent should remove most of the colour from the crystals. Transfer the solid, which is mostly the potassium benzilate salt, to a 125 cm3 round bottomed flask containing 30 cm3 of hot water. Stir the mixture until all the solid has dissolved or until it appears that the remaining solid will not dissolve. If solid still remains in the flask, filter through filter paper using gravity. With stirring, add dropwise 15 cm3 of 1 moldm-3 HCl to the solution of potassium benzilate. The pH should be about 2; if it is higher than this add a few more drops of acid and check the pH again. Allow the mixture to cool to room temperature and then complete the cooling in an ice bath. Let the solid form in the ice bath for at least 30 min, up to about 60 min. If solid has not formed after an hour, you can store your sample until the next lab period. Collect the benzilic acid by vacuum filtration. Wash the crystals with 30-40 mL of water to remove salts. Dry the product and collect the soild.

4.

5.

6.

7.