Author Contributions
Conceptualization, N.-Ł.Z., Z.A., B.T. and Z.-D.M.; Data curation, N.-Ł.Z., Z.A., B.T. and Z.-D.M.; Formal analysis, N.-Ł.Z., Z.A., B.T. and Z.-D.M., Z.M. and H.-B.Z.; Methodology, N.-Ł.Z., Z.A., B.T. and Z.-D.M., Z.M. and H.-B.Z.; Project administration, N.-Ł.-Z.; Visualization, Z.A., B.T. and Z.-D.M., H.-B.Z.; Writing—original draft, N.-Ł.Z., Z.A., B.T. and Z.-D.M., Z.M. and H.-B.Z. Writing—review & editing, N.-Ł.Z., Z.A., B.T. and Z.-D.M., Z.M. and H.-B.Z., W.T. All authors have read and agreed to the published version of the manuscript.
Figure 1.
Extracted ion chromatograms (XIC) obtained for (a) GC, (b) F7, (c) F14 and (d) F28 extracts.
Figure 1.
Extracted ion chromatograms (XIC) obtained for (a) GC, (b) F7, (c) F14 and (d) F28 extracts.
Figure 2.
Kinetics of the absorbance changes in DPPH solutions of green coffee beans’ extract and kombucha ferments. Values are mean of three replicate determinations (n = 3).
Figure 2.
Kinetics of the absorbance changes in DPPH solutions of green coffee beans’ extract and kombucha ferments. Values are mean of three replicate determinations (n = 3).
Figure 3.
The effect of Green coffee extract on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 3.
The effect of Green coffee extract on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 4.
The effect of kombucha ferments’ extracts (F7) on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 4.
The effect of kombucha ferments’ extracts (F7) on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 5.
The effect of kombucha ferments’ extracts (F14) on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 5.
The effect of kombucha ferments’ extracts (F14) on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 6.
The effect of kombucha ferments’ extracts (F28) on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 6.
The effect of kombucha ferments’ extracts (F28) on the DCF fluorescence in fibroblasts and HaCaT cells. The data are expressed as the mean ± SD of 3 independent experiments, each of which consisted of 3 replicates per treatment group.
Figure 7.
Effect of coffee beans’ extract and kombucha ferments on superoxide dismutase activity. Data are the mean ± SD of three independent experiments, in which each concentration was tested in duplicate. **** p < 0.0001, ** p < 0.002.
Figure 7.
Effect of coffee beans’ extract and kombucha ferments on superoxide dismutase activity. Data are the mean ± SD of three independent experiments, in which each concentration was tested in duplicate. **** p < 0.0001, ** p < 0.002.
Figure 8.
The reduction of resazurin after 24 h exposure to the ferments obtained from green coffee beans (1–1000 μg/mL) in cultured (A) fibroblasts (BJ) and (B) keratinocytes (HaCaT). Data are the mean ± SD of three independent experiments, each of which consists of three replicates per treatment group. For BJ, **** p < 0.0001, *** p < 0.0007 versus the control (100%). For HaCaT, **** p < 0.0001 *** p < 0.0008, ** p < 0.003, * p < 0.04 versus the control (100%).
Figure 8.
The reduction of resazurin after 24 h exposure to the ferments obtained from green coffee beans (1–1000 μg/mL) in cultured (A) fibroblasts (BJ) and (B) keratinocytes (HaCaT). Data are the mean ± SD of three independent experiments, each of which consists of three replicates per treatment group. For BJ, **** p < 0.0001, *** p < 0.0007 versus the control (100%). For HaCaT, **** p < 0.0001 *** p < 0.0008, ** p < 0.003, * p < 0.04 versus the control (100%).
Figure 9.
The effect of increasing concentrations of the ferments obtained from green coffee beans (1–1000 μg/mL) on Neutral Red Dye uptake in cultured (A) fibroblasts (BJ) and (B) keratinocytes (HaCaT) after 24 h of exposure. Data are the mean ± SD of three independent experiments, each of which consists of four replicates per treatment group. For BJ, **** p < 0.0001, *** p < 0.0003, ** p < 0.03, * p < 0.01 versus the control (100%). For HaCaT, **** p < 0.0001 ** p < 0.009, * p < 0.02 versus the control (100%).
Figure 9.
The effect of increasing concentrations of the ferments obtained from green coffee beans (1–1000 μg/mL) on Neutral Red Dye uptake in cultured (A) fibroblasts (BJ) and (B) keratinocytes (HaCaT) after 24 h of exposure. Data are the mean ± SD of three independent experiments, each of which consists of four replicates per treatment group. For BJ, **** p < 0.0001, *** p < 0.0003, ** p < 0.03, * p < 0.01 versus the control (100%). For HaCaT, **** p < 0.0001 ** p < 0.009, * p < 0.02 versus the control (100%).
Figure 10.
The effect of increasing concentrations of the ferments obtained from green coffee beans (1–1000 μg/mL) on LDH uptake in cultured (A) fibroblasts (BJ) and (B) keratinocytes (HaCaT) after 24 h of exposure. Data are the mean ± SD of three independent experiments, each of which consists of four replicates per treatment group. For BJ, **** p < 0.0001, *** p < 0.0003, ** p < 0.03 versus the control (100%). For HaCaT, **** p < 0.0001, *** p < 0.009, * p < 0.02 versus the control (100%).
Figure 10.
The effect of increasing concentrations of the ferments obtained from green coffee beans (1–1000 μg/mL) on LDH uptake in cultured (A) fibroblasts (BJ) and (B) keratinocytes (HaCaT) after 24 h of exposure. Data are the mean ± SD of three independent experiments, each of which consists of four replicates per treatment group. For BJ, **** p < 0.0001, *** p < 0.0003, ** p < 0.03 versus the control (100%). For HaCaT, **** p < 0.0001, *** p < 0.009, * p < 0.02 versus the control (100%).
Figure 11.
Collagenase inhibitory activity of coffee beans and kombucha ferments. Data are the mean of three independent experiments, each consisting of two replicates per treatment group. **** p < 0.0001, *** p < 0.0002, ** p < 0.02, * p < 0.01.
Figure 11.
Collagenase inhibitory activity of coffee beans and kombucha ferments. Data are the mean of three independent experiments, each consisting of two replicates per treatment group. **** p < 0.0001, *** p < 0.0002, ** p < 0.02, * p < 0.01.
Figure 12.
Elastase activity of coffee beans and kombucha ferments. Data are the mean of three independent experiments, each consisting of two replicates per treatment group. ** p < 0.03, * p < 0.01.
Figure 12.
Elastase activity of coffee beans and kombucha ferments. Data are the mean of three independent experiments, each consisting of two replicates per treatment group. ** p < 0.03, * p < 0.01.
Figure 13.
Influence of coffee beans’ and kombucha ferments’ extracts on transepidermal water loss (TEWL). Data are the mean ± SD of five independent measurements. *** p < 0.0003, **** p < 0.0001.
Figure 13.
Influence of coffee beans’ and kombucha ferments’ extracts on transepidermal water loss (TEWL). Data are the mean ± SD of five independent measurements. *** p < 0.0003, **** p < 0.0001.
Figure 14.
Influence of coffee beans and kombucha ferments on skin hydration. Data are the mean ± SD of five independent measurements. **** p < 0.0001, *** p < 0.0003, ** p < 0.03.
Figure 14.
Influence of coffee beans and kombucha ferments on skin hydration. Data are the mean ± SD of five independent measurements. **** p < 0.0001, *** p < 0.0003, ** p < 0.03.
Table 1.
Total phenolic (TPC) and flavonoids (TFC) content in green coffee extract and green coffee ferments (DW—dry weight of ferments or extract, GAE—gallic acid, QE—quercetin).
Table 1.
Total phenolic (TPC) and flavonoids (TFC) content in green coffee extract and green coffee ferments (DW—dry weight of ferments or extract, GAE—gallic acid, QE—quercetin).
| TPC (mg GAE/g DW) | TFC (mg QE/g DW) |
---|
GC | 630.05 ± 5.20 a | 156.84 ± 4.11 a |
F7 | 106.76 ± 3.72 b | 17.02 ± 2.33 b |
F14 | 243.14 ± 3.22 c | 51.10 ± 2.84 c |
F28 | 392.56 ± 1.22 d | 66.53 ± 3.84 d |
Table 2.
Polyphenols detected using HPLC-UV-ESI-MS.
Table 2.
Polyphenols detected using HPLC-UV-ESI-MS.
No. | Retention Time (min) | Molecular Formula | Molar Mass (Da) | Precursor Ion [M–H]− m/z | Main Productions MS2 m/z | Identification |
---|
Negative-Ion Mode |
1 | 1.8 | C9H8O4 | 180.2 | 179 [M–H]− | 135 [M-COOH]−, 107 [M-C3H5O2]−, 71 [M-C6H5O2]−, 59 [M-C7H5O2]− | Caffeic acid |
2 | 2.0/2.8 | C7H12O6 | 192.2 | 191 [M–H]− | 127 [M-H-H2O-HCOOH]−, 85 [M-C3H7O4]−, 59 [M-C5H9O4]− | Quinic acid |
3 | 4.1 | C16H18O9 | 354.3 | 353 [M–H]− | 191 [M-3H2O-C6H5O2]−, 179 [M-3H2O-C6H4-COOH]−,135 [M-3H2O-C6H4-C2HO4]− | 3-Caffeoylquinic acid |
4 | 5.2 | C16H18O9 | 354.3 | 353 [M–H]− | 191 [M-3H2O-C6H5O2]−, 179 [M-3H2O-C6H4-COOH]− | 5-Caffeoylquinic acid |
5 | 6.0 | C16H18O9 | 354.3 | 353 [M–H]− | 173 [M-C6H3-2OH-C2H2-COOH]−, 191 [M-3H2O-C6H5O2]−, 179 [M-3H2O-C6H4-COOH]−, 135 [M-3H2O-C6H4-C2HO4]− | 4-Caffeoylquinic acid |
6 | 7.2 | C16H18O8 | 338.3 | 337 [M–H]− | 191 [M-C9H7O11]−, 173 [M-C9H9O3]−, 163 [M-C7H11O5]− | 5-p-Coumaroylquinic acid |
7 | 7.6 | C17H20O9 | 368.3 | 367 [M–H]− | 191 [M-C10H9O3]−, 133 [M-C13H15O4]−, 173 [M-C10H11O4]− | 3-Feruloylquinic acid |
8 | 8.4 | C17H20O9 | 368.3 | 367 [M–H]− | 191 [M-C10H9O3]−, 173 [M-C10H11O4]− | 5-Feruloylquinic acid |
9 | 10.9 | C25H24O12 | 516.4 | 515 [M–H]− | 353 [M-C9H7O3]−, 335 [M-C9H9O4]−, 179 [M-C16H17O8]− | 3,4-Dicaffeoylquinic acid |
10 | 11.5 | C25H24O12 | 516.4 | 515 [M–H]− | 353 [M-C9H7O3]− | 3,5-Dicaffeoylquinic acid |
11 | 12.5 | C25H24O12 | 516.4 | 515 [M–H]− | 353 [M-C9H7O3]−, 179 [M-C16H17O8]− | 4,5-Dicaffeoylquinicacid |
12 | 13.2 | C26H26O12 | 530.5 | 529 [M–H]− | 353 [M-C10H9O3]−, 367 [M-C9H7O3]−, 191 [M-C16H19O8]− | 3-Caffeoyl,5-feruloylquinic acid |
Positive-Ion Mode |
13 | 1.8 | C7H7NO2 | 137.1 | 138 [M + H]+ | 92 [M-H-H-CO2]+, 94 [M-CO2]+, 78 [M-H- H-CO2-CH3]+ | Trigonelline |
14 | 5.2 | C8H10N4O2 | 194.2 | 195 [M + H]+ | 138 [M-CO-N-CH3]+, 110 [M-CO-N-CH3-CO]+ | Caffeine |
Table 3.
Quantification results obtained for GC, F7, F14 and F28 extracts. Values are means ± standard deviation (SD) of triplicate. LOD—limit of detection.
Table 3.
Quantification results obtained for GC, F7, F14 and F28 extracts. Values are means ± standard deviation (SD) of triplicate. LOD—limit of detection.
Compound | Content (mg/100g of Dry Weight of Green Coffee Beans) |
---|
GC | F7 | F14 | F28 |
---|
5-CQA | 4944.9 ± 229.9 | 2486.7 ± 32.3 | 3198.4 ± 15.3 | 2649.0 ± 34.7 |
4-CQA | 562.9 ± 14.7 | 483.7 ± 3.8 | 426.2 ± 6.9 | 531.1 ± 5.5 |
3-CQA | 397.9 ± 13.0 | 390.7 ± 13.0 | 275.5 ± 0.3 | 429.1 ± 1.8 |
3.5-diCQA | 61.2 ± 0.7 | 49.9 ± 4.5 | <LOD | 71.2 ± 2.5 |
4.5-diCQA | 101.2 ± 2.0 | 41.5 ± 3.6 | <LOD | 61.5 ± 2.3 |
3.4-diCQA | 44.0 ± 0.6 | 33.7 ± 2.2 | <LOD | 50.1 ± 1.9 |
3-FQA | 20.7 ± 0.5 | 17.7 ± 2.7 | 9.2 ± 2.4 | 19.0 ± 0.3 |
4-FQA | <LOD | <LOD | <LOD | <LOD |
5-FQA | 544.1 ± 1.3 | 345.8 ± 5.9 | 319.3 ± 4.6 | 350.4 ± 2.7 |
Sum of quantified phenolic compounds | 6677.0 | 3849.7 | 4228.6 | 4161.3 |
Caffeine | 1956.1 ± 29.0 | 1403.4 ± 1.2 | 1165.3 ± 4.1 | 1349.4 ± 1.0 |
Trigonelline | 1205.5 ± 22.1 | 922.3 ± 1.2 | 910.4 ± 1.4 | 839.8 ± 1.0 |
Sum of quantified compounds | 9838.6 | 6175.5 | 6304.4 | 6350.6 |
Table 4.
Ability of green coffee to protect against UV radiation.
Table 4.
Ability of green coffee to protect against UV radiation.
Sample | GC | F7 | F14 | F28 |
---|
SPF | 3.15 ± 0.22 | 0.73 ± 0.09 | 2.14 ± 0.12 | 2.57 ± 0.13 |
Table 5.
Molar extinction coefficients.
Table 5.
Molar extinction coefficients.
Compound | Molar Extinction Coefficient (×104) (M−1 cm−1) | λmax (nm) |
---|
5-CQA | 1.95 | 330 |
4-CQA | 1.80 |
3-CQA | 1.84 |
3,4-diCQA | 3.18 |
3,5-diCQA | 3.16 |
4,5-diCQA | 3.32 |
5-FQA | 1.93 | 325 |
4-FQA | 1.95 |
3-FQA | 1.90 |