Tollens’ Test- Definition, Principle, Procedure, Result, Uses

Tollens’ test is a chemical test used to differentiate reducing sugars from non-reducing sugars. This test is also called the silver mirror test based on the end product of this test.

Objectives of Tollens’ test

  • To distinguish reducing sugars from non-reducing sugars.
  • To detect the presence of aldehyde containing carbohydrates and differentiate them from ketone containing carbohydrates.

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Principle of Tollens’ test

  • The Tollen’s reagent is the alkaline solution of silver nitrate (AgNO3) mixed with liquid ammonia (NH3), which results in the formation of a complex.
  • The aqueous solution of silver nitrate forms a silver aqua complex where the water acts as a ligand.
  • The aqua complexes are then converted into silver oxides (Ag2O) by the action of hydroxide ions.
  • Silver oxide forms a brown precipitate, which is then dissolved by aqueous ammonia resulting in the formation of the [Ag(NH3)2]+ complex.
  • This complex is the primary component of the Tollen’s reagent and is a strong oxidizing agent.
  • The complex then oxidizes the aldehyde group present in some sugars to form a carboxylic acid.
  • At the same time, the silver ions present in the reagent are reduced to metallic silver.
  • The reduction of silver ions into metallic silver results in the formation of a silver mirror on the bottom and sides of the test tube.
  • However, an α-hydroxy ketone gives a positive Tollen’s test as the Tollen’s reagent oxidizes the α-hydroxy ketone into an aldehyde.


2AgNO3 + 2NaOH    →    Ag2O (brown ppt) + 2NaNO3 + H2O

Ag2O (brown ppt) + 4NH3 + 2NaNO3 + H2O    →    2[Ag(NH3)2]NO3 + 2NaOH

Glucose + 2[Ag(NH3)2]NO3 + H2O    →    2 Ag(silver mirror) + 4 NH3 + Gluconic acid + 2 H+



  • Tollen’s reagent: Add 50 ml of 0.1 M AgNO3 to a beaker and to this, add 25 ml of 0.8 M KOH. Now, add sufficient volume of aqueous ammonia in order to dissolve the brown precipitate.
  • Test sample

Materials required

  • Test tubes
  • Test tube stand
  • Pipette


  • Water bath

Procedure of Tollens’ test

  1. Take two clean, dry test tubes and add 1 ml of the test sample in one test tube and 1 ml of distilled water in another as blank.
  2. Add 2 ml of Tollen’s reagent to both the test tubes.
  3. Keep both the test tubes in a water bath for 1 min.
  4. Observe the formation of color and note it down.

Result and Interpretation of Tollens’ test

Tollens' test
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  • The formation of a dark grey precipitate or silver mirror on the bottom and sides of the test tube indicates a positive result, which means that the given sample contains reducing sugars/ aldoses.
  • The absence of such precipitate indicates a negative result, which means that the test sample doesn’t have reducing sugars/ aldoses/ α-hydroxy ketoses.

Uses of Tollens’ test

  • Tollen’s test is routinely performed in chemical laboratories for the qualitative organic analysis, which distinguishes aldehydes from ketones.
  • This test is also used for the differentiation of reducing sugars from non-reducing sugars.

Limitations of Tollens’ test

  • Some carbohydrates that do not have an aldehyde group might give a positive result on Tollen’s test because of the isomerization of such sugars under alkaline conditions.

References and Sources

  1. Tiwari A. (2015). Practical Biochemistry. LAP Lambert Academic Publishing.
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About Author

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Anupama Sapkota

Anupama Sapkota has a bachelor’s degree (B.Sc.) in Microbiology from St. Xavier's College, Kathmandu, Nepal. She is particularly interested in studies regarding antibiotic resistance with a focus on drug discovery.

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