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What is 2,4-Dinitrophenylhydrazine?

The 2,4-Dinitrophenylhydrazine is a chemical compound, C6H3(NO2)2NHNH2, which is also known as DNPH, Borche's reagent and Brady's reagent. The dinitrophenylhydrazine is a red to orange coloured solid. The total compound is a substituted hydrazine and often used qualitatively to test for the carbonyl groups, that are associated with ketones and aldehydes. Also, these hydrazone derivatives can be used as evidence toward the original compound's identity. This derivative's melting point is often used, concerning a database of values, in determining the identity of a particular carbonyl compound. Relatively, it is sensitive to friction and shock; it is also a shock explosive. So, a proper care must be taken towards its use and is usually supplied in wet form to reduce its explosive hazard.

What is a 2,4-Dinitrophenylhydrazine or a 2,4 DNP Test?

The Brady's reagent (the 2,4-dinitrophenylhydrazine) is a red to orange coloured solid, which is usually supplied wet to reduce the explosion risk.

The structure of 2,4-Dinitrophenylhydrazine can be given as follows.

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The derivatives of 2,4-DNP offer a convenient way of separating a mixture of ketones and aldehydes components. Instead of attempting to perform this by the fractional distillation directly, a chemist firstly may take advantage of the easy separation of 2,4-DNP derivatives by column chromatography. Since their formation from ketones and aldehydes is reversible, the separated derivative hydrolysis will regenerate the original carbonyl compounds.

Synthesis of 2,4-Dinitrophenylhydrazine

Although it is not hydrazine that is familiarly substituted, we can make a reasonable inference as hydrazine H2N-NH2 is a good nucleophile. Therefore, the 2,4-Dinitrophenylhydrazine synthesis is done from the product, and it is formed by reacting hydrazine with 2,4-dinitrochlorobenzene. The electron-accepting effect of two nitro groups makes this chloride to displace easily.

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Since the chlorine is ortho, deactivating, and para directing, we should nitrate chlorobenzene. Chlorobenzene is prepared readily from the benzene itself, and so the synthesis is complete, and the reaction can be given below.

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Brady's Reagent

A Brady's reagent can be described as an aqueous solution of 2,4-dinitrophenyl hydrazine (DNP). It reacts with carbonyl compounds (aldehydes & ketone) to produce a coloured precipitate. These resultant precipitates have a sharp melting point. The melting points of these precipitates confirm the carbonyl compounds.

Also, the benzoic acid is used as an antiseptic in medicines, in case of any urinary disorders, and in vapour form for bronchial tubes disinfection.

  • Acetic anhydride reacts with N2O5 to form acetyl nitrate.

(CH3CO)2O + N2O5 → 2CH3COONO2 (Acetyl nitrate)

  • The acid chloride reaction with water decreases with the increase of C-atoms in the alkyl groups.

CH3COCl > CH3-CH2-COCl > CH3-CH2-CH2COCl > …..

Identification of a Carbonyl Compound

The carbonyl compound is mixed with an Brady's reagent acid solution in the methanol compound. These derivatives will result in coloured crystalline solids, known as 2,4-dinitrophenylhydrazones. These resultant crystals are filtered off and purified by the process of re-crystallisation.

The melting temperatures of these derivatives are measured. Then, the original carbonyl compound is identified by comparing it to the tables of melting points of the 2,4-dinitrophenylhydrazine derivatives.

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Brady's reagent can also be used as a test for the presence of a carbonyl compound due to the appearance of the orange crystals, when it is added to either an aldehyde or a ketone.

2,4 DNP Laboratory Test

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  • First, add 5ml of the 2,4-dinitrophenylhydrazine reagent in a test tube

  • Then, add 10 drops of an unknown compound and sharply tap the test tube with a stick or finger to mix. If the crystals do not form immediately, heat in a water bath (60oC) gently for 5 minutes

  • Now, cool the compound in an ice bath until the crystals form. Then, collect the crystals by vacuum filtration with a Hirsch funnel

  • After that, allow the crystals to dry on the Hirsch funnel by drawing the air through crystals. Make a note of the melting point on the Report sheet

  • Usually, the crystals are pure enough to produce a good melting point. Although, recrystallize from a minimum volume of ethanol, if the melting point range is too large.

Reaction with Ethanal

Let us look at the 24 DNP test for aldehydes and ketones or the 24 dnp test for ketones. The alkene ozonolysis gives 24 dnp test positive

The 2,4-Dinitrophenylhydrazine reacts with aldehydes and ketones to further form a 2,4-dinitrophenylhydrazone. For example, with ethanal, it can be given as;

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  • The reaction is a condensation reaction (with the elimination of water). The restaurant product is named with the name of either aldehyde or ketone, followed by a 2,4-dinitrophenylhydrazone. For suppose, ethanal 2,4-dinitrophenylhydrazone.

  • The 2,4-dinitrophenylhydrazine can be used as a process for the identification of aldehydes and ketones. The formed 2,4-dinitrophenylhydrazone is a solid that is purified and the melting point is determined. The comparison of a melting point with a table of known values could identify the compound as aldehyde or ketone

  • All the 2,4-dinitrophenylhydrazone derivatives are orange or yellow solids

  • Often, the solution of 2,4-dinitrophenylhydrazine is called Brady's reagent.

FAQ (Frequently Asked Questions)

1. How Can We Notice the Difference Between Aldehydes and Ketones?

The difference between the aldehyde and ketone is the presence in the aldehyde of a hydrogen atom bound to the double bond of carbon-oxygen. In fact, ketones are not hydrogen. The hydrogen atom presence makes it very simple to oxidize aldehydes. They are also fast reduction agents.

2. If There is a Positive 2/4 DNP Test, What Does it Specify?

The 2,4-Dinitrophenylhydrazine is used for the qualitative identification of aldehyde or ketone functional group carbonyl functionality. The alkene ozonolysis gives 24 DNP test positive, if a positive test is given by the formation of a precipitate, known as a dinitrophenylhydrazone, orange, red, or yellow.

3. Why Does the Glucose Not Offers a 2/4 DNP Test?

This is due to glucose does not react with the 2,4DNP reagent and Schiff's reagent despite having an aldehydrated group. It produces either α-anomer after the internal cycling. There is no free aldehyde group that is present in these forms. Therefore, it does not produce any aldehyde group reaction.

4. Does Alcohol React With Brady's Reagent?

Ketones and aldehydes react to the precipitates of orange, reddish-orange, or yellow with the 2,4-dinitrophenylhydrazine reagent, whereas alcohols do not react. This is one of the successful techniques for checking for the drug existence or demonstrating its absence.