An Efficient Separation Method of Polysaccharides: Preparation of an Antitumor Polysaccharide APS-2 from Auricularia pol
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ORIGINAL
An Efficient Separation Method of Polysaccharides: Preparation of an Antitumor Polysaccharide APS-2 from Auricularia polytricha by Radial Flow Chromatography Jingbo Wang • Xiuping Li • Guanglei Song
Received: 18 October 2012 / Revised: 25 February 2013 / Accepted: 5 March 2013 / Published online: 22 March 2013 Ó Springer-Verlag Berlin Heidelberg 2013
Abstract Auricularia polytricha is an edible mushroom, quite popular in China. Modern pharmacology research indicate that A. polytricha polysaccharides possess antitumor and immunomodulatory activities. In this paper, an antitumor Auricularia polytricha polysaccharide (APS)-2 was purified by radial flow chromatography (RFC) in a column of 500 mL bed-volume (40 cm i.d 9 15 cm) packed with DEAE52-cellulose anion ion-exchange resin. The optimal separation conditions were: sample flowvelocity 15–20 mL min-1, sample volume 160–200 mL, and elution with distilled water and 0.2 M NaCl at a velocity of 35–45 mL min-1. The yield and content of APS-2 obtained under these conditions were 182 mg g-1 total polysaccharides and 98.35 %, respectively. The study provides guidelines for the separation and purification of polysaccharides using a radial flow chromatography column. Keywords Radial flow chromatography Antitumor Polysaccharide Auricularia polytricha
Introduction Mushrooms have recently been identified as an attractive source material for the development of health-care food and drugs due to their various biological activities [1]. Many polysaccharides have been isolated from mushrooms, fungi, yeast, algae, lichens, and plants in recent years and tested for various biological activities [2, 3]. Auricularia polytricha,
which belongs to the Auriculariaceae family, is an edible mushroom, quite popular in China. Modern pharmacology research indicates that A. polytricha polysaccharides possess antitumor and immunomodulatory activities [4–6]. Radial flow chromatography (RFC) and other methods for polysaccharides purification have been reported in recent years [7–11]. However, very little information is available on the of RFC stability for purification of polysaccharides. In a radial flow column, liquid flows radially inside, as compared with axial flow chromatography (AFC) where the direction of liquid flow is along the axis of the column (Fig. 1). Thus, the liquid/buffer travels a shorter distance through the matrix in a radial column as compared with an axial column. RFC can be amplified linearly and provides a fast, effective separation method to handle a large amount of feed phase. It has been used for a variety of industrial applications [12, 13]. APS-2 is a b-D-glucopyranosyl polysaccharide from A. polytricha, having a molecular weight of 2.59 9 105 Da, and it has the highest inhibitory rate of S180 among three A. polytricha polysaccharides isolated in our previous work [14]. The objective of the present investigation was to explore the methods using this fast and effective separation technology and the effects of variables including elution velocity, sample
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