# The Possibility of Using Bee Drone Brood to Design Novel Dietary Supplements for Apitherapy
## 3 Results and Discussion
### Antioxidants  
The analysis of the bioaccessibility of antioxidant compounds was carried out using the FRAP method (Table 4). The capsule containing the only drone brood was found to have the greatest reducing potential, which is due to its highest concentration in the capsule. No reducing potential was found in the ES and CC capsules; therefore, the addition of calcium in both forms reduces the share of drone brood in the capsule by 45.46%, which in turn reduces the antioxidant properties of the undigested fraction, compared to the DB capsule. Taking into account the reduction of the share of drone brood in the DB + ES and DB + CC capsules (up to 54.54% in both cases), a reduction in the reducing potential was found in the DB + ES capsule by 22.36%, and in the DB + CC capsule by 23.90%. The decrease in antioxidant properties in supplements appears to indicate that calcium may block the release of antioxidant compounds or there are interactions between ingredients. After digestion in the stomach and intestines, the FRAP values of all samples  
Figure 4: Increase in free amino acids in drone brood (DB) and designed supplements (DB + ES and DB + CC) during in vitro digestion. decreased, which may be due to the digestion of the compounds. Similar results were obtained by [PERSON] et al. [45], who examined the availability of antioxidant compounds in varietal honeys and showed a decrease in the antioxidant potential in the intestinal fraction compared to the undigested fraction (by up to 33%). It is commonly accepted that antioxidant activity is formed mainly by polyphenols which have anti-inflammatory and anticancer properties [46]. Long-term consumption of foods rich in polyphenols, mainly of plant origin, has been shown to improve conditions of osteoporosis [47]. Due to this, the polyphenol bioaccessibility of the capsules produced was evaluated and is shown in Figure 2d. The content of phenolic compounds in the undigested fraction ranged from 1.97 to 2.28 mg/capsule (for DB + CC and DB, respectively). These compounds were not detected in the prepared eggshell (ES) and calcium carbonate (CC) capsules, which confirms that the drone brood is the only source of phenolic compounds in the designed supplements. The capsule prepared on the basis of brood and eggshells was found to show a 23% higher content of phenolic compounds compared to the DB + CC capsule, which may indicate that calcium carbonate-a synthetic compound-reduces the release of polyphenols from drone brood, whereas compounds derived from eggshells support the release of polyphenols from the matrix. Phenolic compounds were the most bioaccessible in the drone brood (76.31 and 92.98% in the gastric and intestinal fractions, respectively). In addition, the phenolic compounds with DB + ES also showed high bioaccessibility: 60.93 and 52.34% in the gastric and intestinal phases, respectively. However, the recorded values were lower than in the case of DB capsules by 11.33% and 9.37% in the gastric and intestinal fraction. The bioaccessibility of phenolic compounds was conducted in an extended simulation of in vitro digestion for various samples. As the total content of phenolic compounds, the researchers assumed the sum of extractable and hydrolysable fractions, with the hydrolysable fractions being several times higher than the extractable fractions, discussed in this paper as the undigested phase [28; 45]. [PERSON] et al. [45] analyzed the bioavailability of phenolic compounds from varietal honeys using a digestion simulation method analogous to our method. The authors showed that the sum of individual phenolic compounds in the undigested fraction was 46.66% lower compared to the intestinal fraction. The possibility of a food matrix, as well as slight differences in digestion conditions, may have influenced the different behavior between studies. Changes in the polyphenol profile during in vitro digestion of the supplement were confirmed by HPTLC (Figure 5).