Phenol sulphuric acid test

Dissolve 1 g. of the secondary amine in 3-5 ml. of dilute hydrochloric acid or of alcohol (in the latter case, add 1 ml. of concentrated hydrochloric acid). Cool to about 5° and add 4-5 ml. of 10 per cent, sodium nitrite solution, and allow to stand for 5 minutes. Add 10 ml. of water, transfer to a small separatory funnel and extract the oil with about 20 ml. of ether. Wash the ethereal extract successively with water, dilute sodium hydroxide solution and water. Remove the ether on a previously warmed water bath no flames should be present in the vicinity. Apply Liebermann s nitroso reaction to the residual oil or solid thus. Place 1 drop or 0 01-0 02 g. of the nitroso compovmd in a dry test-tube, add 0 05 g. of phenol and warm together for 20 seconds cool, and add 1 ml. of concentrated sulphuric acid. An intense green (or greenish-blue) colouration will be developed, which changes to pale red upon pouring into 30-50 ml. of cold water the colour becomes deep blue or green upon adding excess of sodium hydroxide solution. 

When heated with a strong acid, pentoses and hexoses are dehydrated to form furfural and hydroxymethylfurfural derivatives respectively (Figure 9.20), the aldehyde groups of which will then condense with a phenolic compound to form a coloured product. This reaction forms the basis of some of the oldest qualitative tests for the detection of carbohydrates, e.g. the Molisch test using concentrated sulphuric acid and a-naphthol. 

A reversed-phase liquid chromatographic method was developed for simultaneous determination of carboxylic acids, phenolic compounds, and SA in white wines (84). The diluted samples are injected into a Spherisorb ODS-2 column with a gradient of sulphuric acid (pH 2.5)/methanol as mobile phase. A diode array detector is used, set at 210 nm for carboxylic acids and altered to 278 nm, during the run, for phenolics and SA. The identification of compounds is based on retention time and UV spectra. Some cleanup methods (Sep-Pak C18 and an ion-exchange column) were tested and did not improve the results. The analysis was considered simple, with no sample preparation. Application of this method was illustrated by analyses of Brazilian Welchriesling wines (84). 

Phthalein test. Many phenols yield phthaleins, which give characteristic colorations in alkaline solution, when fused with phthalic anhydride and a little concentrated sulphuric acid. Place in a dry test tube 0.5 g of the compound and an equal bulk of pure phthalic anhydride, mix well together and add 1 drop of concentrated sulphuric acid. Stand the tube for 3-4 minutes in a small beaker of Silicone oil (or paraffin oil) previously heated to 160 °C. Remove from the bath, allow to cool, add 4 ml of 5 per cent sodium hydroxide solution and stir until the fused mass has dissolved. Dilute with an equal volume of water, filter and examine the colour of the filtrate against a white background if the solution exhibits a fluorescence, observe the colour against a black background. 

Place a drop of the test solution in mineral acid on a spot plate, introduce a drop of saturated bromine water followed by 2-3 drops 2m potassium hydroxide (the solution must be alkaline to litmus). Mix thoroughly, add a crystal of phenol, then a drop of the diphenylcarbazide reagent, and finally M sulphuric acid dropwise until the red colour (from the reaction between diphenylcarbazide and alkali) disappears. A blue-violet colouration is obtained. 

Liebermann s Nitroso Reaction.—When phenol in sulphuric acid, phenyl sulphuric acid, is treated with a nitrite or with nitroso amine a daj k green, red or brown color is obtained which changes to blue or green on addition of an alkali. The test is known as Liebermann s nitroso reaction, and may be used in testing for a phenoly a nitrite or a nitroso amine. 

Identification of Phenols.—The reactions of phenols which are of particular value in their identification, are those that take place with alkalies, ferric chloride, and bromine water. Most phenols react with an aqueous solution of sodium hydroxide to form soluble salts, but are insoluble in a solution of sodium carbonate. The behavior of phenols with these two reagents shows their weakly acidic properties, and serves to distinguish them from acids. Phenols which contain strongly negative substituents decompose carbonates, and show all the properties of acids. It is difficult, therefore, to identify as a phenol substances which contain such substituents. Ferric chloride produces marked colorations in aqueous solutions of most phenols. The reagent produces a similar effect with certain other compounds, and the formation of a color with ferric chloride can be taken, therefore, only as an indication of the presence of a phenol. With bromine water most phenols yield a precipitate of a brominated phenol. Other compounds, amines for example, are also converted into insoluble substitution-products by bromine water. Notwithstanding this fact the test is of value. Many phenols form colored products when heated with phthalic anhydride and concentrated sulphuric acid. The reaction will be described under phenolphthalein (558, 639). 

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