A study has been made of the use of 2,6-dichlorophenolindophenol (DCP) and also of 2,4-dinitrophenylhydrazine (DNPH) with and without chromatography of the DNPH derivatives, for the determination of ascorbic acid (AA) and dehydroascorbic acid (DHA) in evaporated milks, and of AA in AA-fortified sterilized milks.
In the DNPH method, when interfering compounds were removed by the chromatographic separation of the DNPH derivatives, absorption curves typical of the pure DHA derivative were obtained.
The DCP method gave erroneously high values for the AA and DHA content, and the lower values obtained by the DNPH method with chromatography of the DNPH derivatives were judged correct. Thus for an evaporated milk, the values for vitamin C content (AA + DHA) were, in mg/100ml reconstituted milk, 0·88 (DCP method), 0·66 (DNPH method without chromatography), and 0·48 (DNPH method combined with chromatography). The AA contents (mg/100 ml) of the 2 fortified sterilized milks were 3·63 and 5·78 (DCP method); 3·01 and 4·28 (DNPH method combined with chromatography).
The AA contents of the 5 different evaporated milks, determined by chromatography of the DNPH derivatives, ranged from 0·07 to 0·63 mg/100 ml reconstituted milk. The DHA content was negligible.
As judged by the shape of the absorption curves after chromatography of the DNPH derivatives, it is concluded that this method is the most reliable of those studied.