Assays by diazotisation

-Nitrous acid(or sodium nitrite in acid) reacts with the primary aromatic amine group to produce a diazonium salt. The reaction is quantitative. Ar-NH2 + HNO2 +HCl Ar-N+N.Cl-

-Amine with the nitrous acid forms nitrosamine, Ar-NH-N=O

-The nitrosamine is followed by tautomerisation & loss of a molecule of water. -The final product is diazonium salt.
-Heterocyclic amines(aminothiazole or aminopyridines) can also be diazotised. -The salts are handled in water. Explosive if isolated or dried. -The method serves as an analytical process via a direct titration of the amine with nitrous acid.

-Nitrous acid solution is unstable. -So that the titration carried out by titrating an acidic solution of the primary aromatic amine with a sodium nitrite solution. -Indicator used is External indicator(starch iodide paper or paste). -After titration, a drop of the titrating solution containing the excess of nitrous acid produces a blue-colour on starch-iodide paste.

-Colour produced should be instantaneous.

KI + HCl HI + KCl 2HI +2HNO2 I2+ 2NO +2H2O

-Liberated iodine reacts with starch to form a blue colour. -Few seconds delay appearance of blue colour indicates the presence of aromatic nitroamine. -Oxidation potential of diazonium salt of nitroamine is so high that excess of nitrous acid can not be determined in the presence of the salt.

-End-point may be detected by amperometrically & potentiometrically(interference or problem by nitroaromatic diazonium salt can be avoided)

Sodium aminosalicylate: Determination of the percentage of C7H6NNaO3.

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175.2g C7H6NNaO3 NaNO2 100ml M 0.01752g C7H6NNaO3 1ml 0.1M NaNO2

Cognate determinations:
Benzocaine,Dapsone,Primaquine phosphate & tablets,Procainamide hydrochloride & injection, Procaine hydrochloride,Sulphacetamide sodium & eye ointment,Sulphadimidine & tablets, Sulphadimidine sodium & injection,Sulphadoxine, Sulphamethizole & tablets,Sulphamethoxazole, Sulphamethoxypyridazine & tablets, Sulphapyridine & tablets,Sulphathiazole & tablets.

Succinylsulphathiazole:Determination of the percentage of C13H13O5N3S2 calculated with reference to the substance dried to constant weight at 105C.

342g C13H13O5N3S2 NaNO2 100ml M 0.0342g C13H13O5N3S2 1ml 0.1M NaNO2

Cognate determination: Succinylsulphathiazole tablets.

Coupling: -The colourless diazonium salt is very reactive. -Reacts with suitable coupling reagent (Ar -H) , e.g. a phenol & aromatic amine. -The product is an azo derivative.
Ar-N N + (Ar -H) Ar-N=N-Ar + H+

-Azo derivatives are coloured & have an absorption maxima at visible region.

-Coupling reagents are 1-naphthol, 2-naphthol & N-(1-naphthyl)-ethylene-1,2-diammonium dihydrochloride.

-The latter reagent results water soluble pink colour solution used in colourimetric assay. -The reagent is also called Bratton-Marshall reagent or NAD reagent.

-Coupling occurs in electron rich position 4 of the reagent. 1)Assay of primary aromatic amine:Sulphonamide etc.

2)Assay of phenol:

References: i) A.H.Beckett/J.B. Stenlake:Practical pharmaceutical chemistry. ii)Keneth A. Connors:A textbook of pharmaceutical analysis. Iii)Takeru higuchi Pharmaceutical analysis.