Fehling’s reaction

C) Reduction of Cupric Ion (Fehling s Reaction). To 4 ml of Fehling s solution (made by mixing equal amounts of copper sulfate solution and a solution of sodium hydroxide containing sodium potassium tartrate ) add a drop of the compound to be tested, and warm. If no change occurs immediately, heat to boiling and set aside. Compare the action of acetaldehyde and acetone or benzal-dehyde and acetophenone. 

Chapter 15 contains a report of a study of the products of the application of Fehling s reaction to free sugars, while their complexation with borate ions is referred to in Chapter 21. 

The nonstoichiometric sugar oxidation process in the presence of alkali is used for both qualitative and quantitative determination of reducing sugars (Fehling s reaction with alkaline cupric tartrate Nylander s reaction with alkaline trivalent bismuth tartrate or using Benedict s solution, in which cupric ion complexes with citrate ion). Hydroxyaldehydes and hydroxyketones are formed by chain cleavage due to retroaldol reaction under nonoxidative conditions using dilute alkali at elevated temperatures or concentrated alkali even in the cold. 

Add 2 -3 drops of phenylhydrazine to about 2 ml. of Fehling s solution in a test-tube and shake the mixture vigorously nitrogen is evolved and reddish-brown cuprous oxide is precipitated. The reaction proceeds rapidly on gentle warming, more slowly in the cold. 

Several variations of the chemical method are in use. In the one described below, a freshly prepared Fehling s solution is standardised by titrating it directly against a standard solution of pure anhydrous glucose when the end-point is reached, I. e., when the cupric salt in the Fehling s solution is completely reduced to cuprous oxide, the supernatant solution becomes completely decolorised. Some difficulty is often experienced at first in determining the end-point of the reaction, but with practice accurate results can be obtained. The titrations should be performed in daylight whenever possible, unless a Special indicator is used (see under Methylene-blue, p. 463). 

Place 10 ml. of 1% starch solution (prepared as described above) in a boiling-tube, add 2 ml. of 1% sodium chloride solution and place the tube in a water-bath maintained at 38-40 . Place about 5 ml. of water in a series of test-tubes and to each add a few drops of 1% iodine solution. Now add 4 ml. of the diluted saliva solution to the starch solution, mix well and note the time. At intervals of about 30 seconds transfer 2 drops of the reacting mixture, by means of a dropping tube, to one of the test-tubes, mix and note the colour. As in the previous experiment, the colour, which is blue at first, changes to blue-violet, red-violet, red-brown, pale brown, and finally disappears at this stage the solution will reduce Fehling s solution. If the reaction proceeds too quickly for the colour changes to be observed, the saliva solution should be diluted. 

Mix each solution well and place the boiling- tubes in a water-bath maintained at 50 . After 10 minutes, transfer i ml. of each of the reaction mixtures to separate test-tubes, add 2 ml. of Fehling s solution to each and boil. A marked reduction is obtained in A, no reduction is obtained in B, and no reduction or only very slight reduction is obtained in C. This shows that the activity of the invertase is destroyed both by heat and alkali. 

Dissolve 0-5 g. of the substance in 10 ml. of 50 per cent, alcohol, add 0-5 g. of solid ammonium chloride and about 0 -5 g. of zinc powder. Heat the mixture to boiling, and allow the ensuing chemical reaction to proceed for 5 minutes. Filter from the excess of zinc powder, and teat the filtrate with Tollen s reagent Section 111,70, (i). An immediate black or grey precipitate or a silver mirror indicates the presence of a hydroxyl-amine formed by reduction of the nitro compound. Alternatively, the filtrate may be warmed with Fehling s solution, when cuprous oxide will be precipitated if a hydroxylamine is present. Make certain that the original compound does not aflfect the reagent used. 

Aromatic aldehydes react with the dimedone reagent (Section 111,70,2). All aromatic aldehydes (i) reduce ammoniacal silver nitrate solution and (ii) restore the colour of SchifiF s reagent many react with sodium bisulphite solution. They do not, in general, reduce Fehling s solution or Benedict s solution. Unlike aliphatic aldehydes, they usually undergo the Cannizzaro reaction (see Section IV,123) under the influence of sodium hydroxide solution. For full experimental details of the above tests, see under Ali-phalic Aldehydes, Section 111,70. They are easily oxidised by dilute alkaline permanganate solution at the ordinary temperature after removal of the manganese dioxide by sulphur dioxide or by sodium bisulphite, the acid can be obtained by acidification of the solution. 

Reaction.—Boil a small quantity for a few minutes with dilute hydrochloric acid, and test with Fehling s solution. The oxime is decomposed into acetone and hydroxylamine,... 

Reaction.—. dA Fehling s solution to benzoin dissolved in alcohol. Benzil is formed and cuprous oxide precipitated. Benzil is also formed on oxidation with nitric acid. 

Symmetrical and unsymmetrical benzoins have been rapidly oxidized to benzils in high yields using solid reagent systems, copper(II) sulfate-alumina [105] or Oxone-wet alumina [105, 106] under the influence of microwaves (Scheme 6.32). Conventionally, the oxidative transformation of a-hydroxy ketones to 1,2-diketones is accomplished by reagents such as nitric acid, Fehling s solution, thallium(III) nitrate (TTN), ytterbium(III) nitrate, ammonium chlorochromate-alumina and dayfen. In addition to the extended reaction time, most of these processes suffer from drawbacks such as the use of corrosive acids and toxic metals that generate undesirable waste products. 

Drug/Lab test interactions False-positive urine glucose reactions may occur with penicillin therapy if Clinitest, Benedict s Solution, or Fehling s Solution are used. It is recommended that enzymatic glucose oxidase tests (such as Clinistix or Tes-Tape) be used. Positive Coombs tests have occurred. High urine concentrations of some penicillins may produce false-positive protein reactions (pseudoproteinuria) with the P.870... 

Reaction with Fehling s (and Benedict s) reagent, aldehydes and ketones (i.e. with sugars - aldoses and ketoses) can reduce Fehling s (and Benedict s) reagents, and they themselves are oxidized. 

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