Identify aldehydes and ketones using Brady’s reagent (2,4-dinitrophenylhydrazine) in this microscale experiment

In this practical, students add various liquid aldehydes and ketones to 2,4-dinitrophenylhydrazine solution in a well-plate to form solid derivatives. They then do the same test with methanol and ethanol, showing that the reaction does not occur with alcohols.

The experiment should take around 15 minutes.



  • Plastic well-plate, 24 wells (eg Sigma ref: M 9655)

For preparation of 2,4-dinitrophenylhydrazine

  • Beaker, 100 cm3



Solutions should be contained in plastic pipettes. See the accompanying guidance on apparatus and techniques for microscale chemistry, which includes instructions for preparing a variety of solutions.

  • Ethanol
  • Propanone
  • p-Methoxybenzaldehyde (or other aromatic aldehyde or ketone)
  • Methanol
  • Ethanal (Acetaldehyde)
  • Solution of 2,4-dinitrophenylhydrazine (see preparation notes below) – requires 24 hours to dissolve completely

For preparation of 2,4-dinitrophenylhydrazine

  • Deionised water

Health, safety and technical notes

  • Read our standard health and safety guidance.
  • Wear eye protection throughout (splash-resistant goggles to BS EN166 3).
  • 2,4-dinitrophenylhydrazine, (NO2)2C6H3NHNH2(s) – see CLEAPSS Hazcard HC030 and CLEAPSS Recipe Book RB033. 2,4-dinitrophenylhydrazine is FLAMMABLE and HARMFUL if swallowed. It is also dangerous in contact with oxidising agents, and can potentially become explosive if allowed to dry out completely. Avoid skin contact.
  • Concentrated sulfuric acid, H2SO4(aq) – see CLEAPSS Hazcard HC098a and CLEAPSS RB098. Concentrated sulfuric acid is CORROSIVE and OXIDISING.
  • Propanone – see CLEAPSS Hazcard HC085A and CLEAPSS Recipe Book RB002. Propanone is highly FLAMMABLE and IRRITANT to eyes and the respiratory system.
  • p-Methoxybenzaldehyde is of low hazard.
  • Methanol – see CLEAPSS Hazcard HC040b. Methanol is highly FLAMMABLE, TOXIC by all routes and causes damage to organs (particularly CNS and optic nerve). Propanol could be used as a safer alternative to methanol. It is FLAMMABLE (see CLEAPPS Hazcard HC084A).
  • Ethanal (acetaldehyde) – see CLEAPSS Hazcard HC034. Ethanal is highly FLAMMABLE, a carcinogen and an eye/respiratory IRRITANT. Protect face when opening containers and ensure laboratory is well-ventilated. Avoid contact with sulfuric acid – violent polymerisation occurs.

Preparing a solution of dinitrophenylhydrazine

  1. Weigh out 0.5 g of 2,4-dinitrophenylhydrazine and dissolve in 10 cm3 of concentrated sulfuric acid in a 100 cm3 beaker. (Leave overnight to allow the compound to dissolve completely.)
  2. Add this solution carefully to a solution of 15 cm3 of ethanol and 5 cm3 of deionised water. This gives the yellow-brown stock solution.

The solution can be used for about 10 days but it gradually deteriorates as a precipitate forms.

This procedure is based on that described in L. Harwood and R. Moody, Experimental organic chemistry – principles and practice, p 242. Oxford: Blackwell Scientific, 1989.


  1. In a well-plate, add 10 drops of 2,4-dinitrophenylhydrazine solution to each of the wells A1–A5 (see diagram below).
  2. Carefully add three drops of ethanal to well A1 (ethanal is very volatile!).
  3. Repeat adding three drops of the other liquids to wells A2–A5.
  4. Observe any changes over the next few minutes.

A diagram showing 24 numbered and lettered circles representing wells in a well-plate

Source: Royal Society of Chemistry

Use this diagram to help you organise the different reactions in this experiment


Compound Observation 
Ethanal  Immediate yellow precipitate 
Propanone  Yellow, crystalline precipitate forms slowly (after 1–2 minutes) 
p -Methoxybenzaldehyde (or other aromatic aldehyde or ketone)  Immediate red precipitate 
Methanol  No change 
Ethanol  No change 

A structural diagram illustrating the reaction between propanone and dinitrophenylhydrazine

Source: Royal Society of Chemistry

Propanone and dinitrophenylhydrazine react to produce propanone 2,4-dinitrophenylhydrazone


  1. Can you write equations for any reactions occurring?
  2. Why do you think that these reactions serve such a useful purpose in identifying aldehydes and ketones?
  3. Aldehydes and ketones will also form derivatives with hydrazine itself. What is the purpose of using 2,4-dinitrophenylhydrazine in this experiment instead of hydrazine?