Zimmermann reagent

The Zimmermann reagent is used as a simple spot-test used in chromatography to presumptively identify alkaloids, especially benzodiazepines, as well as other compounds. It is therefore used in drugs testing.[1][2][3]

It is a two-component reagent, with the first component composed of 1,3-dinitrobenzene (1% w/v) in methanol and the second component composed of 15% potassium hydroxide in water.[4][5][6]

One drop of each component is added to the sample being tested and the resulting colour change is observed to give an indication of the identity of the compound.

The reagent works by forming a reddish-purple Meisenheimer complex at C3 for diazepines with a carbonyl at C2 and an alkyl group at N1.[7] Without these groups it is not possible to form the methylene compound which reacts with dinitrobenzene but triazolo compounds may react.

It is named for the American Biochemist, Robert Zimmermann (b.1937).

Final colors produced by Zimmermann Reagent with various substances
Substance Color
DiazepamReddish purple

See also


  1. Janák, editors, Zdeněk Deyl, Karel Macek, Jaroslav (1975). Liquid Column Chromatography a Survey of Modern Techniques and Applications. Burlington: Elsevier. p. 603. ISBN 9780080858036. Retrieved 10 January 2016.CS1 maint: extra text: authors list (link)
  2. Steroid Dynamics: Proceedings of the Symposium on the Dynamics of Steroid Hormones Held in Tokyo, May, 1965. Academic Press. p. 381. ISBN 9781483270852. Retrieved 10 January 2016.
  3. Callow, R.K.; Campbell, P.N.; Datta, S.P (2013). The Chromatography of Steroids: International Series of Monographs on Pure and Applied Biology: Biochemistry. Elsevier. pp. 232, 270. ISBN 9781483184579. Retrieved 10 January 2016.
  4. "Recommended methods for the I dentification and A nalysis of Barbiturates and Benzodiazepines under International Control" (PDF). UNODC. 2012. p. 25. Retrieved 10 January 2016.
  5. "Rapid Testing Methods of Drugs of Abuse" (PDF). UNODC. 1994. p. 103. Retrieved 10 January 2016.
  6. 0-471-49252-3
  7. Kovar, Karl-Artur & Laudszun, Martina (February 1989). "Chemistry and Reaction Mechanisms of Rapid Tests for Drugs of Abuse and Precursors Chemicals" (PDF). UNODC. p. 13. Retrieved 3 January 2016.
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