Separation and determination of organic acids and phenolic compounds in fruit juices and drinks by high-performance liquid chromatography

A high-performance liquid chromatographic (HPLC) separation method with photo-diode array detection has been developed for the simultaneous determination of organic acids and phenolic compounds in juices and drinks. The chromatographic analysis of organic acids and phenolic compounds was carried out after their elution with sulphuric acid solution (pH 2.5) and methanol from C18 stationary phase. The mobile phase employed was sulphuric acid solution working at a flow-rate of 0.35 ml min−1 for the whole run, while methanol was linearly increased to 0.45 ml min−1 from 15 to 75 min followed by a 5-min isocratic elution. Ten organic acid acids were eluted in 30 min and 21 phenolic compounds, which include phenolic acids and flavonoids, were eluted in the following 50 min. Target compounds were detected at 215 nm. The repeatability (n=3) and between day precision of peak area (n=3) were all within 5.0% RSD. The within-day repeatability (n=3) and between-day precision (n=10) of retention times were within 0.3 and 1.6% relative standard deviation (RSD), respectively. The accuracy of the method was confirmed with an average recovery ranging between 85 and 106%. The method was successfully used to measure a variety of organic acids and phenolic compounds in juices and beverages. This method could also be used to evaluate the authenticity, spoilage or micronutrient contents of juices.