Fluorescence and Phosphorescence Assay of -D Glucans from Basidiomycete Medicinal Mushrooms

Basidiomycete mushrooms contain complex -D-glucans which play an important role in immuno-modulating and anti-tumor activities. The present work involves a novel and intrinsic synchronous fluorescent and phosphorescence assay method for -D-glucans. Synchronous fluorescence and phosphorescence spectroscopy was carried out by a spectrofluorometer in the range of 250 to 750 nm with a  range of 5 -30 nm which exhibited peaks at 492, 540 and 550 nm by using -D-glucan from Euglena gracilis as standard. A micro and high throughput method based on 96-well microtiter plate fluorescence was devised with a excitation and emission of 420 nm and 528 nm, respectively . This assay method presented several advantages over the published colorimetric methods since it is a non-invasive assay method that requires only 0.97 g of -D-glucans in samples, greater sensitivity, speed, assay of many samples and very cheap. -D-glucans of several mushrooms (i.e Poria coccus, Auricularia auricula, Ganoderma lucidium, Pleurotus ostreatus , Cordyceps sinensis , Agaricus blazei, Polyporus umbellatus, Inonotus obliquee) were isolated by using a sequence of several extractions and quantified by either spectrofluorometer or fluorescence microtiter plate reader. 3-D spectra measurements were carried out of -D-glucans from medicinal mushroom strains. FTIR spectroscopy was used to study the structural features ofD-glucans in these mushroom samples.