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The nutritional composition of food for animal production can be enhanced using olive tree and plant by-products due to their high content of bioactive compounds such as pentacyclic triterpenes. Here, we present a novel application of near-infrared spectroscopy (NIRS) for the prediction of the total or individual [maslinic acid (MA), oleanolic acid (OA), and uvaol (UO)] pentacyclic triterpene concentrations in a feed additive obtained from a plant mixture. The oxygen radical absorbance capacity of these types of samples demonstrated the existence of a high antioxidant capacity. The conventional determination methods of pentacyclic triterpene concentration are costly, labor-intensive, and not practical for analyzing several lines within a limited timeframe at the factory level. The optimal regression model developed in our work demonstrated high correlation values for the calibration and validation sets, along with a high residual prediction deviation value. We used 63 samples for the development of the model. The NIRS method can be applied directly to dried powder and makes extraction and high-performance liquid chromatography (HPLC) analysis unnecessary. Our results also demonstrate that NIRS can accurately quantify pentacyclic triterpenes even at low concentrations in food additives. It can be used at the factory level to directly determine the pentacyclic triterpene concentrations in the additive powder at the same time that the powder is produced.