Page 392 - IJB-10-6

International Journal of Bioprinting Innovative manufacturing of ω-3-enriched chocolate

increased (p < 0.05) compared to the control sample. Tolve by Martini et al., who examined dark chocolate. After
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et al. reported that the highest polyphenol concentration the intestinal phase, a significant increase (p < 0.05) was
31
was observed in the control (spreadable chocolate: 369 observed in the TPC and antioxidant capacity of all the
mg GAE/100 g); spreadable chocolate enriched with 11.6 chocolate samples (Figure 8). These increases could be due
g/kg Mg-CaCO nanoparticles and 166 μg/kg vitamin D to enhanced extractability of phenolics and flavonoids,
3
exhibited the lowest value (272 mg GAE/100 g). Caponio influenced by factors such as pH, enzymes, organic solvent
et al. noted that the TPC values of artisanal (Choco-A) dependency, bile concentration, and incubation (3 h)
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31
and industrial (Choco-I) dark chocolate were 20.66 and during the intestinal phase. 32,62 In a study by Tolve et al.,
21.53 mg GAE/g, respectively. the bioaccessibility of polyphenols in chocolate hazelnut
spreads was examined after simulated in vitro digestion; the
Similarly, the antioxidant capacities of ω-3-coated and
mixed chocolate samples were also significantly increased authors reported that the same trend may occur for all the
samples, in which there is a high polyphenol concentration
(p < 0.05) compared to the control sample. The DPPH in the simulated salivary phase, followed by a reduction in
values of chocolate samples ranged from 34.85 to 40.11 the simulated gastric phase, and a subsequent increase in
μmol TE/g dw; the ABTS values of chocolate samples the simulated intestinal phase. The TPC and antioxidant
ranged from 25.67 to 30.80 μmol TE/g dw. As indicated capacities of ω-3-coated and mixed chocolate samples
by the antioxidant capacity results, differences among were also significantly increased (p < 0.05) compared
DPPH, ABTS, and CUPRAC assays were observed; higher to the pure chocolate sample after in vitro digestion.
values were observed using the CUPRAC method. These Ingredients of the ω-3-coated and mixed chocolate
variations could be related to the variety of reaction samples may influence the structure and degradation
conditions used in the study, such as the wavelength and of the matrix when exposed to digestion. Particle size,
reaction time at which the measurements were obtained. amount, distribution of fat, emulsifier type, quantity, and
In the present study, the CUPRAC values of all chocolate solid particles all affect the chocolate microstructure.
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extracts were higher than the values obtained using the In the present study, the highest TPC and antioxidant
DPPH and ABTS methods. The Cu ion takes part in capacity were noted in chocolate coated with ω-3-SA MP
2+
forming free radicals; the reduction of the cupric ion before in vitro digestion. In contrast, the highest TPC and
indicates a mechanism other than that associated with the antioxidant capacity were found in ω-3-mixed chocolate
DPPH and ABTS methods, which reflects the antioxidant samples after in vitro digestion. Despite the high initial
potential. This observation can be associated with the fact TPC and antioxidant capacity in the chocolate coated with
that the CUPRAC assay may measure the hydrophilic ω-3-SA MP, the BI value was not the highest among the
and lipophilic antioxidant capacities of the extracts, as chocolate samples (Figure 8). The BI values of the TPC
the reagent is soluble in organic and aqueous solvents. 32,60 were 67%, 62%, and 94% in pure chocolate, ω-3-coated,
Caponio et al. demonstrated that the ABTS and DPPH and ω-3 mixed, respectively, while the BI values of DPPH,
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values of artisanal chocolate (Choco-A) were 98.39 and ABTS, and CUPRAC ranged between 69–98%, 32–60%,
79.81 µmol TE/g, respectively; the ABTS and DPPH values and 81–97%, respectively.
of industrial (Choco-I) dark chocolate were 88.99 and
68.65 µmol TE/g, respectively. 3.10. In vitro release study
Encapsulation techniques are used for the controlled release
3.9. In vitro digestion test of bioactive compounds and to increase bioavailability. To
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The changes in TPC and antioxidant capacity values compare the controlled release characteristics, in vitro ω-3
during in vitro gastrointestinal digestion (oral, gastric, and release studies were conducted using chocolates coated with
intestinal phases) are provided in Figure 8. For all chocolate ω-3-SA MP and mixed with ω-3. To mimic gastrointestinal
samples, an increase was noted from the oral phase to the fluids, ω-3 release profiles of chocolates were performed in
intestinal phase. Compared to the initial amount, the lower PBS (pH 4 and 7) at 37°C; the results are displayed in Figure
values of TPC and antioxidant capacity in the oral phase 10. The UV spectra for the 0.2–1 µg/mL ω-3 concentration
(Figure 8) may be justified by the low solubility of the range are displayed in Figure 10A. The released ω-3 was
compounds in salivary fluid and the short period (3 min). identified at an absorbance of 214 nm. Figure 10C and D
The TPC of chocolate samples in the oral phase was noted demonstrate the cumulative release of ω-3 from ω-3-SA-
to vary between 137.65 mg GAE/100 g dw and 217.29 coated chocolate and ω-3-mixed chocolates at pH 4 and 7.
mg GAE/100 g dw. During the in vitro simulation of GI As observed in Figure 10C, 59% burst release was observed
digestion, an increase was noted in the total antioxidant in the first 15 min from SA-ω-3-coated chocolates at pH
capacity of the chocolate samples from the oral phase to 7, and the release was completed in 1 h. A burst release
the gastric phase. Similar results have been described of 27% was observed in the first 1 h from SA-ω-3-coated

Volume 10 Issue 6 (2024) 384 doi: 10.36922/ijb.3969