In the present study an HPLC-DAD method was developed for the determination of the main carotenoid, zeaxanthin dipalmitate in the fruits of Lycium barbarum. Aim was to develop and optimize an extraction protocol to allow fast, exhaustive and repeatable extraction, suitable for the labile carotenoid content. Use of liquid N2 allowed the grinding of the fruit. A step of ultrasonication with water removed efficiently the polysaccharides and enabled the exhaustive extraction of carotenoids by hexane:acetone 50:50. The assay was fast, simple and permitted the quality control of a large number of commercial samples including fruits, juices and a jam. The HPLC method was validated according to ICH guidelines and satisfied the requirements. Finally the overall method was validated for precision (% R.S.D ranging between 3.81 and 4.13) and accuracy in three concentration levels. The recovery was between 94% and 107% with R.S.D. values < 2%, within the acceptable limits, especially if the difficulty of the matrix is taken into consideration.