目的: 建立桑黄多糖多重指纹图谱法,为桑黄质量评价提供参考。方法: 采用高效凝胶色谱法（HPGFC-RID法）,色谱柱为TSK-GEL®G3000 PWXL（7.8 mm×30 cm,7 μm）,以20 mmol · L-1 醋酸-醋酸钠缓冲液（pH 5.7）为流动相,流速0.5 mL · min-1,进样量15 μL,柱温35 ℃,RID检测器;采用红外光谱法（FT-IR法）,测定范围为4 000~400 cm-1,分辨率4 cm-1,扫描16次;采用HPLC-UV糖谱法,色谱柱为Agilent 5 HC-C18（2） （250 mm×4.6 mm,5 μm）,以乙腈-0.02 mol · L-1乙酸铵（20 ∶ 80）为流动相,流速1 mL · min-1,检测波长250 nm,进样量10 μL,柱温35 ℃,UV检测器。建立桑黄多糖多重指纹图谱,对29批桑黄多糖的重均分子量、特征吸收官能团、单糖组成进行比较,并结合化学计量学和主成分分析（PCA）,分析其种内及种间差异。结果: HPGFC-RID糖谱分析显示,桑树桑黄（SH）、杨树桑黄（YH）、暴马桑黄（BH）和松木层孔菌（松树桑黄） （SSH）多糖均有2个主要色谱峰（P1、P2）。对于主要色谱峰P1,BH多糖的重均分子量最高,其次依次为SSH、SH和YH多糖;对于主要色谱峰P2,YH多糖的重均分子量最高,其次依次为SSH、SH和BH多糖。FT-IR糖谱分析表明,SH、YH、BH和SSH多糖具有相似的红外光谱吸收峰,种内、种间差异不明显,但袋料栽培与椴木栽培的桑黄多糖在1 800~900 cm-1范围内主要吸收峰强度不同。HPLC-UV糖谱结合化学计量学分析显示,桑黄多糖由甘露糖、鼠李糖、葡萄糖、木糖和岩藻糖组成,其中SH、YH（椴木栽培）、BH和SSH多糖中葡萄糖含量最高,YH（袋料栽培）多糖中木糖含量最高。各单糖组成分布结果表明,不同品种桑黄单糖含量存在明显差异。此外,PCA将YH（袋料栽培）聚为一类,SH、YH（椴木栽培）、BH和SSH聚为一类,说明栽培方式的不同也会影响桑黄的质量。结论: 桑黄多糖多重指纹图谱分析方法可对不同品种桑黄质量进行有效评价,为桑黄质量控制奠定基础。
Objective: To establish a multiple fingerprint analysis method for polysaccharides from Sanghuangporus sanghuang, providing a reference for quality evaluation. Methods: 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). Results: 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. Conclusion: The multiple fingerprint analysis method for Sanghuangporus polysaccharides effectively evaluates the quality of different Sanghuangporus varieties, providing a foundation for quality control.
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