目的 构建黄精炮制品多糖的酶水解产物指纹图谱，结合化学计量学方法评价不同蒸晒次数黄精炮制品多糖的差异。方法 采用糖苷酶水解，结合高效薄层色谱（high thin-layer chromatography，HPTLC）、亲水作用色谱-高效液相色谱-蒸发光检测器（hydrophilic interaction liquid chromatography-high performance liquid chromatography-evaporative light scattering detector，HILIC-HPLC-ELSD）和超高效液相色谱-四级杆飞行时间串联质谱（ultra high pressure liquid chromatography-quadrupole time of flight tandem mass spectrometry，UHPLC-QTOF/MS）对相应的酶解产物进行高效分离和检测。由此产生的自下而上的指纹图谱反映了黄精炮制品多糖的变化，通过相似度和偏最小二乘-判别分析（partial least squares-discriminant analysis，PLS-DA）对指纹图谱进行综合评价。结果 HPTLC分析发现，果糖随着蒸晒次数的增加逐渐降低，结合色谱分析发现，六蒸六晒和十二蒸十二晒为黄精炮制过程中的转折点。同时根据酶水解中寡糖的变化，利用相似度和PLS-DA对不同蒸晒次数的黄精炮制品进行了清晰的聚类。结合UHPLC-QTOF/MS分析表明，不同蒸晒次数黄精炮制品中含有蔗糖、蔗果三糖、蔗果四糖、蔗果五糖、蔗果六糖、蔗果七糖及蔗果八糖。结论 基于酶解法分析不同蒸晒次数样品的质量动态变化，结合所建立的多元指纹图谱，进一步阐释了不同蒸晒次数黄精炮制品的科学内涵。

Objective To construct the fingerprint of the enzymatic hydrolysis products of Huangjing (Polygonati Rhizoma, PR) to evaluate the difference in polysaccharides from processed PR with various steaming periods combined with chemometrics. Methods Glycosidase hydrolysis combined with high thin-layer chromatography (HPTLC), hydrophilic interaction liquid chromatography-high performance liquid chromatography-evaporative light scattering detector (HILIC-HPLC-ELSD) and ultra high pressure liquid chromatography-quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF/MS) were used to efficiently separate and detect the corresponding enzymatic hydrolysis products. The resulting bottom-up fingerprint reflected the changes in polysaccharides of processed PR, and the fingerprint was comprehensively evaluated through similarity and partial least squares-discriminant analysis (PLS-DA).Results With longer heating and drying times, fructose steadily was decreased, according to HPTLC examination. Combining with chromatographic analysis, it was discovered that the PR’s processing reached a turning point at six-steaming-six-basking and twelve-steaming-twelve-basking. Using similarity and PLS-DA, clear clustering was done on the processed PR with varying steaming and drying periods based on the changes in oligosaccharides during enzymatic hydrolysis. With the aid of UHPLC-QTOF/MS analysis, it was discovered that processed PR with various steaming periods contains fructo-oligosaccharide DP8, sucrose, 1-kestose, nistose trihydrate, 1F-fructofuranosylnystose, kestohexaose, and 1-kestoheptaose. Conclusion The quality dynamic changes of samples with different steaming and drying times based on enzymatic examination combined with multivariate fingerprint elucidated the scientific connotation of the processed PR with distinct steaming and drying times.

安徽省药品监督管理局监管科学研究重点项目——基于中药糖谱法黄精多糖质量控制标准研究（AHYJ-KJ-202308）