Abstract
Four different extraction methods, including hot water extraction (HWE), ultrasonic-assisted extraction (UAE), enzyme-assisted extraction (EAE) and ultrasonic-enzyme-assisted extraction (UEAE), were applied to extract polysaccharides from Trifolium repens L. (TRPs). In addition, response surface methodology (RSM) was performed to optimize the extraction conditions of TRPs. The results showed that different extraction methods had significant effects on the extraction yields and antioxidant activities of TRPs. TRPs extracted by the EAE method (10.57%) and UEAE method (10.62%) had significantly higher extraction yields than TRPs extracted by the HWE method (8.35%) and UAE method (9.43%) (P < 0.05). However, there was no significant difference between the extraction yields of the EAE method and UEAE method (P > 0.05). TRPs extracted by the EAE method had a higher content of uronic acid and exhibited better antioxidant capacities. Therefore, EAE was selected as the optimal extraction method to extract TRPs. The optimal extraction conditions of EAE to extract TPRs were liquid–solid ratio 30 mL/g, enzymolysis time 87 min, enzyme-complex dosage 1.6% and pH 6, leading to a TRPs yield of 13.15%.
Highlights
The Trifolium genus contains approximately 240 species of clovers[1], mainly distributed in western North America, the Mediterranean basin and the highlands of eastern Africa[2]
The uronic acid contents were significantly different in the following order: hot water extraction (HWE)-Trifolium repens L. (TRPs) (4.04%) < ultrasonic-assisted extraction (UAE)-TRPs (4.27%) < UEAETRPs (5.17%) < enzyme-assisted extraction (EAE)-TRPs (5.42%) (P < 0.05)
There were no significant differences on the extraction yields and uronic acid contents between the EAE method and ultrasonic-enzyme-assisted extraction (UEAE) method (P > 0.05)
Summary
The Trifolium genus contains approximately 240 species of clovers[1], mainly distributed in western North America, the Mediterranean basin and the highlands of eastern Africa[2]. H. et al conducted an experiment to study the optimal water extraction condition for polysaccharides from white clover using a single-factor experiment and an L9 (34) orthogonal experiment[10]. To the best of our knowledge, research on the physicochemical characteristics and the bioactivities of polysaccharides from Trifolium repens L. Response surface methodology (RSM) can be used to obtain the optimal extraction conditions of polysaccharides, while Box-Behnken design (BBD) is more efficient compared to other methods due to fewer runs[18]. The physicochemical properties and antioxidant activities of TRPs were determined to select the suitable extraction method. The final purpose of this study is to provide a basis for the development and utilization of Trifolium repens L
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