CHROMATOGRAPHY, Vol. 30 (2009), No. 1, pp. 25-35
Investigation on the Evaluation Method of Fungi
-Polysaccharide Marker Substance for the Identification by Gel Permeation Chromatography-
Takashi Nakamura1, Hiroyuki Nishi2 and Kazuaki Kakehi3
1Analytical Chemistry Research Department, CMC Research Center, Mitsubishi Tanabe Pharma Corporation, 3-16-89 Kashima, Yodogawa-ku, Osaka 532-8505, Japan
2School of Pharmacy, Yasuda Women's University, 6-13-1 Yasuhigashi, Asaminami-ku, Hiroshima 731-0153, Japan
3School of Pharmacy, Kinki University, 3-4-1 Kowakae, Higashiosaka-shi, Osaka 577-8502, Japan
Polysaccharides of five fungi (two fungi crude drugs such as Polyporus umbellatus and Poria cocos, three fungi, Ganoderma lucidum, Grifola frondosa and Agaricus blazai, used for functional health foods, were evaluated by employing gel permeation chromatography (GPC) with a special column, which is applicable under the alkaline conditions, and multi angle laser light scattering (MALLS) and refractive index (RI) detectors. Total polysaccharides content by the color reaction using phenolic sulfonic acid was also investigated. Three different extraction methods of polysaccharides, with hot water, hot alkali solution or cold alkali solution, were applied for the sample preparation. As a result, different extraction method resulted in the different GPC pattern and polysaccharide content. Polysaccharide component was detected in all three extraction methods from all five fungi by employing ultra filtration (UF). Especially large amount of polysaccharides was detected in cold alkali extraction from Poria cocos. GPC patterns under the different extraction conditions were revealed to be useful for the identification and/or evaluation of fungi. These different results seem to be due to the difference in the sugar conformation of each fungus. Results obtained by iodine.starch reaction and β-glucan specific reaction also indicated polysaccharides structural differences in each fungi extracts in the viewpoint of amount (frequency) and chain length of β-1, 3-glucan and α-1, 4-glucan structure. It was found that polysaccharides, extracted by three different extraction methods with UF membrane, could be a marker substance for the identification and evaluation method of fungi.
Keywords: fungi, polysaccharide, marker substance, identification, crude drug, gel permeation chromatography, multi angle light scattering detection.