To establish a multiple fingerprint analysis method for polysaccharides from Sanghuangporus sanghuang, providing a reference for quality evaluation.

High-performance gel filtration chromatography(HPGFC-RID) was used with a TSK-GEL^®G3000 PWXL column (7.8 mm×30 cm, 7 μm). The mobile phase was 20 mmol·L^-1 HAc-NaAc buffer (pH 5.7) at a flow rate of 0.5 mL·min^-1. The injection volume was 15 μL, column temperature was 35 ℃, and detection was carried out using a RID detector. Fourier transform infrared(FT-IR) spectroscopy was performed in the range of 4 000 to 400 cm^-1 with a resolution of 4 cm^-1 and 16 scans. HPLC-UV monosaccharide profiling was done using an Agilent 5 HC-C₁₈ column (250 mm×4.6 mm, 5 μm), with acetonitrile-0.02 mol·L^-1 ammonium acetate (20：80) as the mobile phase, at a flow rate of 1 mL·min^-1, detection wavelength of 250 nm, an injection volume of 10 μL, and column temperature of 35 ℃. An UV detector was used for detection. Multiple fingerprints of Sanghuangporus polysaccharides were established, and the mass average molar mass, characteristic absorptive functional groups, and monosaccharide compositions of 29 batches of Sanghuangporus polysaccharides were compared and analyzed for their intraspecific and interspecific variations by combining with chemometrics and principal component analysis (PCA).

HPGFC-RID monosaccharide profiling revealed that the polysaccharides from Sanghuangporus sanghuang (SH), Sanghuangporus vaninii (YH), Sanghuangporus baumii (BH), and Phellinus pini (SHH) all exhibited two major chromatographic peaks (P1, P2). For P1, BH polysaccharides had the highest molecular weight, followed by SSH, SH, and YH polysaccharides. For P2, YH polysaccharides had the highest molecular weight, followed by SSH, SH, and BH polysaccharides. FT-IR analysis indicated that SH, YH, BH, and SSH polysaccharides shared similar infrared absorption peaks, with no significant differences between species or within species. However, the intensity of the main uptake peaks in the range of 1 800-900 cm^-1 was different between bagged and linden cultivated Sanghuangporus polysaccharides. HPLC-UV monosaccharide profiling combined with chemometrics revealed that Sanghuangporus polysaccharides consist of mannose, rhamnose, glucose, xylose, and fucose. Glucose content was the highest in SH, YH (linden cultivation), BH, and SSH polysaccharides, while xylose was the highest in YH (bagged cultivation). Monosaccharide composition varied significantly among different Sanghuangporus varieties. PCA grouped YH (bagged cultivation) as one class, and SH, YH (linden cultivation), BH, and SSH as another, indicating that cultivation methods also influence Sanghuangporus quality.

The multiple fingerprint analysis method for Sanghuangporus polysaccharides effectively evaluates the quality of different Sanghuangporus varieties, providing a foundation for quality control.