Semi-Continuous Subcritical Water Extraction of Flavonoids from Citrus unshiu Peel: Their Antioxidant and Enzyme Inhibitory Activities
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Preparation
2.2. Chemicals
2.3. Subcritical Water Extraction (SWE)
2.4. Methanol Extraction
2.5. Acid and Base Hydrolysis
2.6. HPLC Analysis
2.7. Response Surface Design
2.8. Antioxidant Activity Measurement
2.9. Enzyme Inhibitory Activity Measurement
2.10. Statistical Analysis
3. Result and Discussion
3.1. Composition of Flavonoids in C. unshiu Peel
3.2. Optimization of SWE Process
3.3. Effects of Extraction Parameters on Individual Flavonoid Yields
3.4. Comparison of Flavonoid Compositions in the SW Extract and Acid and Base Hydrolysis
3.5. Antioxidant and Enzyme Inhibitory Activities in the SW Extracts
3.6. Correlations between Flavonoid Yields and Functional Properties of the SW Extracts
3.7. Antioxidant Activities of Individual Citrus Flavonoids
3.8. Enzyme Inhibitory Activities of Individual Citrus Flavonoids
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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No | X1 | X2 | Hesperidin and Its Hydrolysis Products (μg/g Dry Sample) | Narirutin and Its Hydrolysis Products (μg/g Dry Sample) | Polymethoxyflavones (μg/g Dry Sample) | Total Flavonoids (μg/g Dry Sample) | Total Soluble Solids (%) | Total Flavonoid Concentration in the Extract (mg/g Dry Extract) | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Hesperidin | Hesperetin-7-O-glucoside | Hesperetin | Narirutin | Prunin | Naringenin | Sinensetin | Nobiletin | Tangeretin | ||||||
1 | 150 | 1 | 24,700.4 | N.D. | N.D. | 5442.7 | N.D. | N.D. | 13.7 | 68.8 | 29.0 | 30,253.5 | 45.2 ± 0.3 d | 66.6 |
2 | 150 | 2 | 37,995.1 | N.D. | N.D. | 8040.6 | N.D. | N.D. | 18.4 | 85.4 | 45.5 | 46,184.3 | 56.0 ± 1.7 b,c | 82.4 |
3 | 170 | 1 | 30,498.0 | 1640.8 | 1591.0 | 5631.8 | 237.8 | 222.3 | 13.2 | 76.4 | 37.4 | 39,947.9 | 58.2 ± 2.4 a,b | 68.5 |
4 | 170 | 2 | 42,975.4 | 756.8 | 937.1 | 8027.2 | 200.4 | 145.7 | 15.7 | 90.6 | 49.6 | 53,197.9 | 57.9 ± 1.5 a,b | 91.8 |
5 | 145 | 1.5 | 27,935.1 | N.D. | N.D. | 6687.7 | N.D. | N.D. | 15.6 | 76.5 | 37.5 | 34,751.6 | 38.5 ± 1.1 e | 90.1 |
6 | 175 | 1.5 | 34,749.8 | 2048.0 | 1802.7 | 6595.3 | 326.0 | 268.1 | 14.2 | 82.2 | 39.1 | 45,924.3 | 58.1 ± 0.7 a,b | 79.0 |
7 | 160 | 0.75 | 24,345.7 | 698.2 | 433.6 | 4848.1 | 162.5 | 102.3 | 11.1 | 62.4 | 28.2 | 30,691.3 | 45.3 ± 1.2 d | 67.6 |
8 | 160 | 2.25 | 44,617.3 | 370.6 | 642.5 | 8760.0 | N.D. | 135.2 | 17.7 | 91.2 | 54.5 | 54,688.7 | 60.6 ± 2.2 a | 90.1 |
9 | 160 | 1.5 | 36,858.1 | 757.8 | 870.2 | 6856.2 | 141.4 | 126.1 | 15.9 | 82.5 | 41.6 | 45,749.5 | 55.1 ± 0.6 c | 82.9 |
10 | 160 | 1.5 | 36,786.7 | 701.2 | 856.7 | 6832.4 | 135.9 | 127.7 | 16.6 | 83.9 | 40.8 | 45,580.8 | --- | 75.1 |
11 | 160 | 1.5 | 37,486.7 | 739.4 | 884.1 | 6762.1 | 142.1 | 135.1 | 15.9 | 83.2 | 42.1 | 46,289.9 | --- | 76.3 |
Flavonoid Content (μg/g Dry Sample) | |||
---|---|---|---|
Subcritical Water Extract * | Acid Hydrolysate | Base Hydrolysate | |
Hesperidin | 34,749.8 | 12,081.2 ± 1188.3 | 16,124.1 ± 491.9 |
Hesperetin-7-O-glucoside | 2048.0 | 2462.4 ± 96.0 | N.D. |
Hesperetin | 1802.7 | 2204.0 ± 39.9 | N.D. |
Narirutin | 6595.3 | 1240.7 ± 54.2 | 1620.8 ± 8.3 |
Prunin | 326.0 | 703.9 ± 23.6 | N.D. |
Naringenin | 268.1 | 863.0 ± 34.5 | N.D. |
Sinensetin | 14.2 | 18.3 ± 0.6 | 15.4 ± 0.3 |
Nobiletin | 82.2 | 91.1 ± 0.7 | 74.3 ± 1.7 |
Tangeretin | 39.1 | 57.6 ± 2.5 | 41.8 ± 1.1 |
Total | 45,924.3 | 19,722.2 ± 1236.3 | 17,876.4 ± 483.9 |
X1 | X2 | Antioxidant Activity | Enzyme Inhibition Activity | |||||
---|---|---|---|---|---|---|---|---|
DPPH Radical Scavenging Activity (mg AAE/g Dry Sample) | FRAP (mmol FSE 100/g Dry Sample)) | ORAC (mg TE/g Dry Sample) | Xanthine Oxidase (%) | ACE (%) | α-Glucosidase (%) | Pancreatic Lipase (%) | ||
145 | 1.5 | 8.3 ± 0.1 f | 24.7 ± 1.1 f | 169.5 ± 5.2 d | 11.6 ± 1.3 f | 12.8 ± 1.5 f | 12.8 ± 1.1 f | 52.0 ± 1.9 g |
150 | 1 | 8.2 ± 0.3 f | 24.9 ± 0.9 f | 122.8 ± 5.8 e | 16.6 ± 1.4 e | 14.2 ± 0.6 e, f | 16.5 ± 0.8 e | 59.8 ± 1.9 e, f |
150 | 2 | 14.8 ± 0.5 c | 28.9 ± 0.7 e | 338.8 ± 16.6 b | 20.8 ± 1.8 d | 22.5 ± 1.3 d | 21.4 ± 1.4 d | 66.8 ± 2.0 d |
160 | 0.75 | 9.5 ± 0.6 e | 25.5 ± 0.9 f | 281.3 ± 17.5 c | 15.3 ± 1.0 e | 16.2 ± 0.7 e | 20.7 ± 1.2 d | 58.0 ± 1.6 f |
160 | 1.5 | 13.3 ± 0.7 d | 45.3 ± 1.3 c | 316.2 ± 26.3 b, c | 21.4 ± 0.8 d | 21.6 ± 0.5 d | 23.1 ± 1.1 d | 64.1 ± 2.7 d, e |
160 | 2.25 | 18.6 ± 0.3 a | 48.8 ± 1.3 b | 397.4 ± 21.8 a | 30.9 ± 1.7 c | 35.6 ± 1.5 b | 41.0 ± 1.7 b, c | 73.3 ± 1.8 c |
170 | 1 | 14.0 ± 0.0 c, d | 39.3 ± 1.1 d | 327.6 ± 5.0 b | 32.3 ± 1.8 c | 32.6 ± 1.7 c | 39.2 ± 2.1 c | 78.0 ± 1.4 b |
170 | 2 | 17.2 ± 0.4 b | 44.4 ± 1.4 c | 423.4 ± 34.9 a | 38.1 ± 1.5 b | 35.5 ± 1.2 b | 42.1 ± 1.2 b | 81.3 ± 3.1 b |
175 | 1.5 | 18.6 ± 0.4 a | 52.6 ± 1.8 a | 424.9 ± 32.5 a | 52.6 ± 2.5 a | 48.9 ± 2.6 a | 55.5 ± 2.4 a | 86.8 ± 3.4 a |
Antioxidant Activity | Enzyme Inhibition Activity (IC50) | ||||||
---|---|---|---|---|---|---|---|
DPPH Radical Scavenging Activity (mg AAE/g) | FRAP (mmol FSE 100/g) | ORAC (mg TE/100 mg) | Xanthine Oxidase (mg/L) | ACE (mg/L) | α-Glucosidase (mg/L) | Pancreatic Lipase (mg/L) | |
Hesperidin | 45.3 ± 0.9 c | 399.7 ± 21.1 c | 247.3 ± 13.2 f | >2000 | 1375.8 ± 62.1 b | >2000 | 418.4 ± 21.7 a |
Hesperetin-7-O-glucoside | 51.6 ± 1.4 b | 439.8 ± 18.4 b | 323.9 ± 13.7 d | >2000 | 106.5 ± 3.8 f | 1395.5 ± 69.9 b | 84.4 ± 4.2 d |
Hesperetin | 115.8 ± 3.9 a | 549.1 ± 21.1 a | 710.9 ± 12.0 a | 275.4 ± 17.7 c | 358.2 ± 7.8 d | 131.8 ± 7.3 e | 104.6 ± 5.2 d |
Narirutin | 17.6 ± 1.2 e | 102.6 ± 4.1 e | 283.4 ± 15.7 e | 1104.7 ± 40.7 a | 1,937.1 ± 42.5 a | 1004.0 ± 42.6 c | >2000 |
Prunin | 26.5 ± 0.2 d | 99.5 ± 3.6 e | 363.9 ± 18.1 c | 862.5 ± 36.6 b | 221.2 ± 6.2 e | 622.7 ± 28.0 d | >2000 |
Naringenin | 52.6 ± 3.3 b | 150.3 ± 3.7 d | 475.1 ± 13.3 b | 129.3 ± 6.4 d | 459.7 ± 15.0 c | 72.2 ± 3.8 e | >2000 |
Sinensetin | N.D. | N.D. | 7.6 ± 0.4 g | >2000 | 364.0 ± 13.4 d | >2000 | 301.4 ± 22.2 b |
Nobiletin | N.D. | N.D. | 6.0 ± 0.4 g | >2000 | 322.5 ± 4.7 d | >2000 | 426.4 ± 27.6 a |
Tangeretin | N.D. | N.D. | 5.8 ± 0.4 g | >2000 | 352.0 ± 8.9 d | 1763.1 ± 66.6 a | 148.7 ± 6.5 c |
Positive control | 116.7 ± 3.7 d (Allopurinol) | 6.9 ± 0.2 g (Captopril) | 1366.9 ± 73.2 b (Acarbose) | 54.4 ± 3.6 e (Orlistat) |
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Kim, D.-S.; Lim, S.-B. Semi-Continuous Subcritical Water Extraction of Flavonoids from Citrus unshiu Peel: Their Antioxidant and Enzyme Inhibitory Activities. Antioxidants 2020, 9, 360. https://doi.org/10.3390/antiox9050360
Kim D-S, Lim S-B. Semi-Continuous Subcritical Water Extraction of Flavonoids from Citrus unshiu Peel: Their Antioxidant and Enzyme Inhibitory Activities. Antioxidants. 2020; 9(5):360. https://doi.org/10.3390/antiox9050360
Chicago/Turabian StyleKim, Dong-Shin, and Sang-Bin Lim. 2020. "Semi-Continuous Subcritical Water Extraction of Flavonoids from Citrus unshiu Peel: Their Antioxidant and Enzyme Inhibitory Activities" Antioxidants 9, no. 5: 360. https://doi.org/10.3390/antiox9050360