Comparison of different extraction methods for polysaccharides from Dendrobium officinale stem
Introduction
The extraction of polysaccharides from a plant source needs to follow a series of procedures including solvent infiltration, polysaccharide dissolution and diffusion (Fig. 1A). The improvement of solvent penetration into cells and solute diffusion are usually the key factors to facilitate the extraction progress (Chen, Meng, Zhang, & Liu, 2009). Hot water extraction method (HWE) could easily accelerate the diffusion rate of polysaccharides and improve the extraction efficiency with the increased temperature of water. The extraction process can also be further facilitated by physical methods. Ultrasonic-assisted hot water extraction (UHWE) (Zhang, Li, Xiong, Jiang, & Lai, 2013), microwave-assisted hot water extraction (MHWE) (Tao & Xu, 2008) and enzyme-assisted hot water extraction (EHWE) (Pan, Wang, Ye, Zha, & Luo, 2015) were commonly used and proved to be more efficient, as they can largely promote the dissolution of polysaccharides through biodegradation or mechanical destruction of plant cell walls. Recently, freeze-thawing cold-pressing (FTCP) method has been favored, because the plant cell wall can be effectively destructed, thus the polysaccharide dissolution rate are high (Guo, Liu, Bai, & Yang, 2013). In addition, this method can avoid the degradation of polysaccharides caused by heat or ultrasound (Zhang, Chen et al., 2013), and retain good bioactivity.
Dendrobium officinale (Orchidaceae) is a valuable Chinese herbal medicine for its polysaccharide (DOP). It has showed a variety of pharmacological properties, such as antioxidative (Huang et al., 2016; Luo, Ge, Fan, Chun, & He, 2011), immune-stimulating (Cai et al., 2015), hypoglycemic (Zhao, Son, Kim, Jang, & Lee, 2007) and anti-tumor activities. There is a growing number of research and increasing demand of the DOP-based supplements and Chinese patent medicines. HWE (Huang et al., 2016), UHWE (Fan, Luo, Luo, & Chun, 2009) and MHWE (Liu & Xiong, 2009) have been used in previous studies for the extraction of polysaccharides from Dendrobium officinale, but these research did not specify the best extract method for DOP. Thus, study of the efficiency of different extraction methods can potentially benefit the relevant pharmaceutical industry by providing evidence-based information for improvement of industrial equipment and materials.
Detailed structural studies of Dendrobium officinale revealed that the neutral polysaccharide was dominantly composed of β-D-glucopyranose and β-D-mannopyranose, both of which possess hydroxyl radical scavenging activity and thus anti-oxidative capacity (Xing, Cui, Nie, Phillips, Goff & Wang, 2013). Oxidative stress has been proved to be implicated in aging (Remmen & Richardson, 2001), neurodegeneration (Deng, Thompson, Gao, & Hall, 2007), diabetes mellitus (Wolff, Jiang, & Hunt, 1991), cardiovascular disease (Siti, Kamisah, & Kamsiah, 2015), and cancer (Reuter, Gupta, Chaturvedi, & Aggarwal, 2010). Antioxidants might be useful in combating with these processes and diseases. In current study, the antioxidant capacity after different extraction method can be used as an indicator to determine the remained amounts of active polysaccharides.
The present study was aimed to evaluate and screen the most suitable extraction method for DOP by comparing the outcomes of DOP’s yields of different extraction methods, including hot water extraction (HWE), cold-pressing (CP), freeze-thawing cold-pressing (FTCP), ultrasonic-assisted hot water extraction (UHWE), microwave-assisted hot water extraction (MHWE) and enzyme-assisted hot water extraction (EHWE). Then, DOP samples were characterized by chemical analysis, gas chromatography (GC), high-performance gel permeation chromatography (HPGPC), Fourier transform infrared spectoscopy (FT-IR) and scanning electron microscopy (SEM). Finally, the antioxidant activities in vitro of DOP samples were evaluated and compared to find the most bioactive sample.
Section snippets
Materials and reagents
Dendrobium officinale stems was provided by Guangdong Yongshengyuan Biological Technology Co., Ltd. (Guangdong, China). Dendrobium officinale stems used in present study was 3-year-old and collected in May of the third year after planting. Their quality was identified by Prof. Danyan Zhang from Guangzhou University of Chinese medicine. They were cultivated on the GAP base of Raoping County, Guangdong Province.
Dextrans with different molecular weight, 2,2′-azino-bis (3- ethylbenzothiazoline-
Effects of different extraction methods on the yield of DOP
In this study, six extraction methods were used to extract DOP. The yield of DOP with HWE, CP, FTCP, UHWE, MHWE and EHWE were shown in Fig. 1, and the LSD analysis results were shown in Table 1. The yield of total polysaccharides by UHWE was comparatively higher, and this was consistent with most of the findings in previous studies (Yan et al., 2015). But there was no significant difference between FTCP and UHWE. This may due to the processes of frequent freezing and fast thawing, which
Conclusion
Effects of different extract method (CP, FTCP, HWE, UHWE, MHWE and EHWE) on the yield, characteristic and antioxidant activity of DOP were investigated in this study. The results showed that DOP extracted by different methods were composed mainly of mannose and glucose. UHWE and FTCP had the highest yield, while DOPUHWE showed the lowest antioxidant activity, and DOPFTCP showed the superlative antioxidant activity. The differences of the yields and antioxidant activities were not significant in
Acknowledgements
This work was partly supported by the National Natural Science Foundation of China (No. 81503387), Hong Kong, Macao and Taiwan Special Science and Technology Cooperation Program (No. 2014DFH30010), Six Talent Peaks Project in Jiangsu Province (2017-YY-003), Qing Lan Project of Jiangsu Province, Science and Technology Program of Guangdong (No. 2013B090800052), Guangdong Provincial Science and Technology Projects (No. 2013A022100002, 2014A030304059), Application Science and Technology Research
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These authors contributed equally to this paper.