The proposed method for ascorbic acid (Vitamin C) (AA) determination is based on the oxidation of AA to dehydroascorbic acid with a Cu(lI)-2,9-dimethyl-1, 10-phenanthroline (neocuproine (Nc)) reagent in ammonium acetate-containing medium at pH7, where the absorbance of the formed bis(Nc)-copper(l) chelate is measured at 450 nm. This chelate was formed immediately and the apparent molar absorptivity for AA was found to be 1.60 x 10(4) dm(3) mol(-1) cm(-1). Beer ' s law was obeyed between 8.0 x 10(-6) and 8.0 x 10(-5) M concentration range. The relative standard deviation for 90 mu g AA was 3%. The Cu(Il)-Nc reagent is a milder and therefore more selective oxidant than the conventional Fe(III)1, 10-phenanthroline (phen) reagent used for the same assay. This feature makes the proposed method superior for real samples such as fruit juices containing weak reductants such as citrate, oxalate and tartarate that otherwise produce positive errors in the Fe(III)-phen method when equilibrium is achieved. The developed method was applied to a number of commercial fruit juices, pharmaceutical preparations containing Vitamin C, and red wine. The meta-bisulfite content of wine was removed with an anion exchanger at pH 3 prior to analysis, and a difference extractive-spectrophotometric method of AA assay in wine was developed so as to suppress the interferences caused by wine anthocyanins and polyphenols. The findings of the developed method for fruit juices and pharmaceuticals were also statistically compared with those of HPLC so as to establish it as a reliable novel method. (c) 2004 Elsevier B.V. All rights reserved.