BENEDICTS TEST

INTRODUCTION PROCEDURE RESULTS && DISCUSSION APPLICATIONS

reduction of metallic ions by sugars

Reducing sugars are oxidized by copper (II) ions in two other saccharide test solutions: Benedict's reagent , a mildly basic solution and Barfoed's reagent, a mildly acidic solution. The presence of red copper (I) oxide precipitate indicates that the saccharide has reduced the copper (II) ions.

introduction

One liter of Benedict's solution contains: 173 grams sodium citrate 100 grams sodium carbonate 17.3 grams cupric sulfate pentahydrate. Benedict's solution or Benedict's test Stanley Rossiter Benedict.

introduction

BENEDICT'S REAGENT IS USED AS A TEST FOR THE PRESENCE OF REDUCING SUGARS.

BENEDICT'S TEST WILL DETECT THE PRESENCE OF ALDEHYDES (EXCEPT AROMATIC ONES), AND ALPHA-HYDROXY-KETONES, INCLUDING THOSE THAT OCCUR IN CERTAIN KETOSES.

introduction

1. Take 3 mL of Benedicts solution in a test tube and boil vigorously for about one minute 2. Add 8 drops of test solution and continue boiling for another 2 minutes.

procedure

PROPOSED RESULT :
During a water bath, which is usually 4 10 minutes, the solution should progress in the colors of blue (with no glucose present), green, yellow, orange, red, and then brick red or brown (with high glucose present).

results &&Discussion

PROPOSED RESULT :
Experiment Observation Inference

Substance in water + 3ml Benedict's solution, then boil for few minutes and allow to cool.

Red ppt or green ppt or yellow ppt obtained Substance in water + 3ml Solution Benedict's solution, then boil for remains clear few minutes and allow to cool.

Reducing sugar e.g. Glucose is present Reducing sugar is not present

results &&Discussion

Galactose Glucose

(+) (+)

Orange Red Solution Blue-Red Solution

Fructose Maltose Sucrose Xylose

(+) (+) (-) (+)

Dark Brown with Brick Red precipitate Dark Brown with Brick Red precipitate Bluish solution Brick Red Solution

results &&Discussion

results &&Discussion

SHOWS POSITIVE TEST FOR:

REDUCING SUGARS

REACTIONS:

REDUCING SUGARS are oxidized by the copper ion in solution to form a carboxylic acid and a reddish precipitate of copper (i) oxide.
KETOSE FRUCTOSE is strictly not a reducing sugar and gives a POSITIVE TEST .

results &&Discussion

 A colour change would signify the presence of glucose. The common disaccharides lactose and maltose are directly detected by benedict's reagent, because each contains a glucose with a free reducing aldehyde moiety, after isomerization.

Ketose fructose is an alpha-hydroxy-ketone, and gives a positive test because it is converted to the ldoses glucose and mannose by the base in the reagent.

results &&Discussion

 Sucrose contains two sugars (fructose and glucose) joined by their glycosidic bond in such a way as to prevent the glucose isomerizing to aldehyde, or the fructose to alpha-hydroxy-ketone form.
Sucrose is thus a non-reducing sugar which does not react with benedict's reagent.

results &&Discussion

Sucrose indirectly produces a positive result with Benedict's reagent if heated with dilute hydrochloric acid prior to the test, although after this treatment it is no longer sucrose.

The acidic conditions and heat break the glycosidic bond in sucrose through hydrolysis. The products of sucrose decomposition are glucose and fructose, both of which can be detected by benedict's reagent, as described above.

results &&Discussion

results &&Discussion

Benedict's test uses a mixture of copper (II) sulfate, sodium citrate, and sodium carbonate in a mildly basic solution.

If the saccharide is a reducing sugar, it will reduce the copper (II) ions to copper (I) oxide, a red precipitate .

results &&Discussion

Alkaline solutions of copper are reduced by sugars having a free aldehyde or ketone group. the citrate will form soluble complex ions with Cu++, preventing the precipitation of CuCO3 in alkaline solutions.

results &&Discussion

1) DIABETES URINE TEST - BENEDICT'S TEST

This is a very simple and effective method of ascertaining the presence or the amount of glucose in the urine and can be done by the diabetic himself.

Perform the test two hours after a meal. In the initial stages of the disease, a diabetic does not lose sugar in his urine, when on empty stomach. Hence if the benedict's test is performed in the fasting state, it is possible to miss the diagnosis of the disease. Even if sugar is detected in the urine by benedict's test, the diagnosis of diabetes should be confirmed by blood-analysis.

applications

APPARATUS : BENEDICT'S SOLUTION DROPPER, TEST-TUBE, TEST-TUBE HOLDER. PROCEDURE : 1. TAKE 5 ML (ONE TEASPOON) OF BENEDICT'S SOLUTION IN THE TEST -TUBE.

2. HOLDING THE TEST-TUBE WITH THE HOLDER, HEAT IT OVER A SPIRIT LAMP TILL T HE BENEDICT'S SOLUTION BOILS WITHOUT OVERFLOWING.
3. DROP 8 TO 10 DROPS OF URINE INTO THE BOILING BENEDICT'S SOLUTION. 4. AFTER AGAIN BOILING THE MIXTURE, LET IT COOL DOWN. 5. WHILE COOLING, THE MIXTURE CHANGES COLOUR.

6. THE COLOUR OF THE MIXTURE SERVES AS A GUIDE TO THE AMOUNT OF SUGAR I N THE URINE : BLUE-SUGAR ABSENT; GREEN-0.5% SUGAR; YELLOW-1% SUGAR; ORANGE1.5% SUGAR; BRICK RED-2 % OR MORE SUGAR.

applications

2)BENEDICT'S TEST ON BEER.

Beer contains mainly water and some but apart from these substances a large number of other biomolecules. These molecules [including derivatives of ethanol and higher alcohols like 2 -methylbutanol] are there because they are synthesized during the beer production process or are being produced during storage. Staling of beer, resulting in a bad taste is due to the deterioration of some of the components naturally present in beer. These reactions are initiated by light and catalyzed by e.G. Riboflavin, one of these components in beer. Ultimately the beer contains degradation products like aldehydes, ketones, [furfural, acetone, trans -2-butenal and trans-2-nonenal] oxidized lipids, and so on. The reason that beer gives a positive a positive reaction in the benedict test is that the beer contains these compound that react with the reagent. These compound may be remaining sugar and the other components like aldehydes and ketones.

applications

Le ga sp i , G. A . E sse nt ials o f b io ch e mist ry. 2 0 0 9 F rie d m a n , p . J. Bio ch e mist ry 5 th e d . 1 9 9 5 . Litt le , b ro wn an d co mp any. Usa D e n n isto n , k. J.; To p p in g , J. J.; Caret , R.L. Ge n e ral, o rgan ic an d b io ch e mist ry 5 th e d . M cgraw- h ill e d u cat io n a sia Be n e d ict 's te st fo r re d u cin g su gars. Ret rie ve d o n 1 2 ju ly 2 0 1 1 . Ret rie ve d at htt p: / / www2 .Vo lstate .Edu/ CHE M /1 0 3 0 / labs/carbo 9 .Ht m S e liwa n o ff 's te st - so lu t io n . Ret rie ve d o n ju ly 1 2 , 2 0 1 1 . Ret rie ve d at htt p: / /scie n ce . Jrank.Org / page s/ 1 9 2 6 3 /se liwan o ff's - te st .Ht ml S e liwa n o ff 's te st fo r keto n e h exo se s. Ret rie ve d o n ju ly 1 2 , 2 0 1 1 . Ret rie ve d at htt p: / / www2 .Vo lstate .Edu/ CHE M /1 0 3 0 / labs/carbo 9 .Ht m l S ch re ck , J.O., Lo ff re do, W.M . Qu alitat ive te st in g fo r carb o hyd rate s. 2 0 0 9 . Ret r ie ve d 1 2 j u ly 2 0 1 1 f ro m htt p: / / www.Ce rlab s.Co m/exp e rime nts/ 1 0 8 7 5 4 0 4 4 6 4 .Pd f

references