Benedict's reagent is used as a test for the presence of all monosaccharides, and generally also reducing sugars. These include glucose, galactose, mannose, lactose and maltose. Even more generally, Benedict's test will detect the presence of aldehydes (except aromatic ones), 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..
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. The mixture is heated in a boiling water bath, and any precipitate formed is recorded as a positive result for the presence of reducing sugars in the food.
The common disacharrides 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 with 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 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.
- Benedict, S. R. (1908). "A Reagent For the Detection of Reducing Sugars". J. Biol. Chem. 5: 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.