Benedict's reagent is used as a test for the presence of reducing sugars. This includes all monosaccharides and many disaccharides, including lactose and maltose. Even more generally, Benedict's test will detect the presence of aldehydes, and alpha-hydroxy-ketones, including those that occur in certain ketoses. Thus, although the ketose fructose is not strictly a reducing sugar, it is an alpha-hydroxy-ketone, and gives a positive test because it is converted to the aldoses glucose and mannose by the base in the reagent.
A positive test with Benedict's reagent is shown by a colour change from clear blue to a brick-red precipitate. One litre of Benedict's reagent can be prepared from 100 g of anhydrous sodium carbonate, 173 g of sodium citrate and 17.3 g of copper(II) sulfate pentahydrate. It is often used in place of Fehling's solution.
Benedict's reagent contains blue copper(II) ions (Cu2+) which are reduced to copper(I) ions (Cu+). These are precipitated as red copper(I) oxide which is insoluble in water. Benedict's Reagent provides a quantitative test for reducing sugars along with qualitative test. The color of the obtained precipitate gives an idea about the quantity of sugar present in the solution. A greenish precipitate indicates about 0.5% concentration; yellow precipitate indicates 1% concentration; orange indicates 1.5% and red indicates 2% or higher concentration.
To test for the presence of monosaccharides and reducing disaccharide sugars in food, the food sample is dissolved in water, and a small amount of Benedict's reagent is added. 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). 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.
Sucrose (table sugar) 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. 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.
Starches do not react or react very poorly with Benedict's reagent, due to the relatively small number of reducing sugar moieties, which occur only at the ends of carbohydrate chains. Inositol (myo-inositol) is another carbohydrate which produces a negative test.
Benedict's reagent can be used to test for the presence of glucose in urine. Glucose found to be present in urine is an indication of diabetes mellitus. Once a reducing sugar is detected in urine, further tests have to be undergone in order to ascertain which sugar is present. Only glucose is indicative of diabetes.
Benedict's solution, deep-blue alkaline solution used to test for the presence of the aldehyde functional group, - CHO. The substance to be tested is heated with Benedict's solution; formation of a brick-red precipitate indicates presence of the aldehyde group. Since simple sugars (e.g., glucose) give a positive test, the solution is used to test for the presence of glucose in urine, a symptom of diabetes. One liter of Benedict's solution contains 173 grams sodium citrate, 100 grams sodium carbonate, and 17.3 grams cupric sulphate pentahydrate. It reacts chemically like Fehling's solution; the cupric ion (complexed with citrate ions) is reduced to cuprous ion by the aldehyde group (which is oxidized), and precipitates as cuprous oxide, Cu2O.
|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||Reducing sugar e.g. Glucose is present|
|Substance in water + 3ml Benedict's solution, then boil for few minutes and allow to cool.||Solution remains clear or is a little blue||Reducing sugar is not present|
Benedict's quantitative reagent is used to determine how much reducing sugar is present. This solution forms as white precipitate rather than a red one and so can be used in a titration. The titration should be repeated with 1% glucose solution instead of the sample for calibration.
Other oxidizing reagents
Other reducing reagents (opposite)
- Benedict, S. R. (1 January 1909). "A Reagent For the Detection of Reducing Sugars". J. Biol. Chem. 5 (6): 485–487.
- Fehling's Test for Reducing Sugars
- Robert D. Simoni, Robert L. Hill, and Martha Vaughan (2002). "Benedict's Solution, a Reagent for Measuring Reducing Sugars: the Clinical Chemistry of Stanley R. Benedict". J. Biol. Chem. 277 (16): 10–11. doi:10.1074/jbc.M110516200. PMID 11773074.