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. 2015 Aug 26;20(9):15550-71.
doi: 10.3390/molecules200915550.

Optimization of Ultrasonic-Assisted Extraction of Flavonoid Compounds and Antioxidants from Alfalfa Using Response Surface Method

Chang-Liang Jing  1 , Xiao-Fang Dong  2 , Jian-Ming Tong  3
Affiliations

Optimization of Ultrasonic-Assisted Extraction of Flavonoid Compounds and Antioxidants from Alfalfa Using Response Surface Method

Chang-Liang Jing et al. Molecules. .

Abstract

Ultrasonic-assisted extraction (UAE) was used to extract flavonoid-enriched antioxidants from alfalfa aerial part. Response surface methodology (RSM), based on a four-factor, five-level central composite design (CCD), was employed to obtain the optimal extraction parameters, in which the flavonoid content was maximum and the antioxidant activity of the extracts was strongest. Radical scavenging capacity of the extracts, which represents the amounts of antioxidants in alfalfa, was determined by using 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) methods. The results showed good fit with the proposed models for the total flavonoid extraction (R² = 0.9849), for the antioxidant extraction assayed by ABTS method (R² = 0.9764), and by DPPH method (R² = 0.9806). Optimized extraction conditions for total flavonoids was a ratio of liquid to solid of 57.16 mL/g, 62.33 °C, 57.08 min, and 52.14% ethanol. The optimal extraction parameters of extracts for the highest antioxidant activity by DPPH method was a ratio of liquid to solid 60.3 mL/g, 54.56 °C, 45.59 min, and 46.67% ethanol, and by ABTS assay was a ratio of liquid to solid 47.29 mL/g, 63.73 °C, 51.62 min, and 60% ethanol concentration. Our work offers optimal extraction conditions for total flavonoids and antioxidants from alfalfa.

Keywords: alfalfa; antioxidant capacity; flavonoid compounds; response surface methodology; ultrasonic-assisted extraction.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of liquid to solid ratio on the extraction yield of total flavonoids (a); ABTS (b) and DPPH (c) radical-scavenging capacities of extracts from alfalfa.
Figure 2
Figure 2
Effects of extraction temperature on the extraction yield of total flavonoids (a); ABTS (b) and DPPH (c) radical-scavenging capacities of extracts from alfalfa.
Figure 3
Figure 3
Effects of extraction time on the extraction yield of total flavonoids (a); ABTS (b) and DPPH (c) radical-scavenging capacities of extracts from alfalfa.
Figure 4
Figure 4
Effects of ethanol concentration on the extraction yield of total flavonoids (a); ABTS (b) and DPPH (c) radical-scavenging capacities of extracts from alfalfa.
Figure 5
Figure 5
Response surface plots (3D) showing the effects of variables on total flavonoid extraction. (a) liquid to solid ratio and temperature; (b) liquid to solid ratio and time; (c) liquid to solid ratio and Ethanol concentration; (d) time and temperature; (e) temperature and ethanol concentration; (f) time and ethanol concentration.
Figure 6
Figure 6
Response surface plots (3D) showing the effects of the extraction parameters on ABTS radical-scavenging capacity of alfalfa extracts. (a) liquid to solid ratio and temperature; (b) liquid to solid ratio and time; (c) liquid to solid ratio and Ethanol concentration; (d) time and temperature; (e) temperature and ethanol concentration; (f) time and ethanol concentration.
Figure 7
Figure 7
Response surface plots (3D) showing the effects of the extraction parameters on DPPH radical-scavenging capacity of alfalfa extracts. (a) liquid to solid ratio and temperature; (b) liquid to solid ratio and time; (c) liquid to solid ratio and Ethanol concentration; (d) time and temperature; (e) temperature and ethanol concentration; (f) time and ethanol concentration.
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