Fresh meat preparations, as fresh sausages, hamburger, minced meats etc, are susceptible to rapid oxidation and the consequent browning of meat red color compromises the product attractiveness. As a result for the producers, there is the drastic reduction of products shelf-life. In order to remedy such economic loss, the addition of some substances with antioxidant effect may represent a valid solution. However, the addition of some food antioxidants (i.e. nicotinic acid, sulphiting agents, etc.) is not admitted in fresh meat preparations by the actual Normative; moreover, for other substances, such as ascorbic acid, some use restriction subsist. In this work, an analytical method for the determination of two food antioxidants, such as ascorbic acid and nicotinic acid, in fresh meat preparations, by high performances liquid chromatography coupled with UV Diode Array Detection, is described. The reliability of this method was assured by developing a full validation procedure, by following the actual European Guidelines. The most important validation parameters, such as linearity, specificity, accuracy, detection and quantification limits, ruggedness and measurement uncertainty were evaluated, resulting conform with European requirements. Considering that ascorbic acid and nicotinic acid are subject to use restriction in fresh meat preparations, this method may be considered a valid tool in food inspection for organisms in charge of food controls.