The alphaPROX assay: fluorescence screening of the inhibitory effects of hydrophilic antioxidants on protein oxidation.

We report on a new and convenient high-throughput fluorescence technique for determining antioxidant capacities of hydrophilic food samples. The new method is called alphaPROX (anti protein oxidation) and is based on an equimolar complex of diphenylhexatriene propionic acid (DPHPA) and bovine serum albumin (BSA) in aqueous buffer at pH 7.4. DPHPA is a reporter fluorophore that becomes nonfluorescent upon free radical-induced oxidation. In a typical assay, the DPHPA/BSA complex is challenged with peroxyl radicals and shows almost the same susceptibility to oxidation as unlabeled BSA. The progress of protein oxidation and its inhibition by antioxidants at physiological pH is determined from the time-dependent decrease in DPHPA fluorescence intensity. The alphaPROX method was compared to other techniques frequently used to measure antioxidant capacities. In this article, representative results are provided for the inhibitory effects of pure food components, fruit juices, wines, and various polar plant extracts on protein oxidation.

An improved method to discover adulteration of Styrian pumpkin seed oil.

Pumpkin seed oil is rather expensive compared to other vegetable oils. Therefore, it is often adulterated by the addition of cheaper oils. In contrast to other edible oils, the content of Delta 5-sterols is very low, while Delta 7-sterols are dominating. The determination of Delta 5-sterols, especially beta-sitosterol has proven to be a good possibility to detect admixture of cheap vegetable oils to a level below which the economic profit is not significant any more. The presented method is a variant of an analysis concept, which has been published previously in European Food Research and Technology by Mandl et al. [Eur. Food Res. Technol. 209 (1999) 400. ]. It includes saponification of the triglycerides as a first step followed by separation of the potassium salts of the fatty acids from the unsaponifiable fraction by adsorption chromatography. In order to enhance gas chromatographic properties of the analytes, the hydroxyl function of the sterols is derivatized with N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) to the trimethylsilylether. Finally, the analytes are separated on a capillary column of medium polarity (HP 35 MS) in a temperature programmed run within 18 min. Detection of the analytes was done by flame ionization. Special attention was set onto the precision and repeatability of the method.