目的 探究单一酶法与复合酶法提取黄精Polygonati Rhizoma多糖的最优条件；分析黄精九蒸九晒过程中多糖组分含量的变化，为黄精和其他中药材的炮制方法提供相应理论依据。方法 响应面（RSM）优化酶法提取黄精多糖，1-苯基-3-甲基-5-吡唑啉酮（PMP）柱前衍生法和HPLC法对九蒸九晒的姜形黄精、蒸出液中的多糖进行分析，建立黄精古法炮制以及蒸出液的多糖特征图谱。比较不同酶对黄精中多糖提取效率的影响。结果 单一酶中纤维素酶提取效果最好，4种复合酶中纤维素酶与果胶酶结合提取效果最好；通过RSM优化得到最佳提取条件：纤维素酶在酶解温度为51℃、酶的添加量为2%、酶解pH值为5.40的情况下，精制黄精多糖提取物中多糖总量为（675.34±0.10）mg/g；PMP-HPLC检测炮制过的4种黄精，均含有甘露糖、核糖、鼠李糖、葡萄糖、半乳糖。其中鸡头黄精Polygonatum sibiricum、姜形黄精P.cyrtonema、竹节黄精P.odoratum多糖均以葡萄糖为主，马鞭黄精P.filipes多糖以甘露糖为主。随着姜形黄精被炮制次数的增加，各单糖含量先增加后下降再增加后趋于稳定，以甘露糖、半乳糖变化明显。蒸出液各单糖含量先升高后趋于稳定，其中甘露糖、葡萄糖的变化明显。结论 研究表明纤维素酶参与的单一酶法和复合酶法提取黄精多糖，不仅可以提升黄精多糖的提取效率，也提高黄精多糖纯度；九蒸九晒会影响黄精中的多糖提取得率；PMP-HPLC特征图谱能够准确地表征黄精炮制过程中多糖含量的变化，可用于评价黄精及炮制药材的多糖组分的差异。

Objective To investigate the best extracting conditions of Polygonatum sibiricum polysaccharides by single and compound enzymatic methods, and to study on the change content of polysaccharide in processing of P. sibiricum by steamed and shined for nine times, which provide theoretical basis for processing of P. sibiricum and other Chinese herbal medicines. Methods Response surface method (RSM) was used to optimize the extraction of polysaccharides in P. sibiricum. PMP precolumn derivatization and HPLC characteristic map technology were used to identify and analyze the changes of polysaccharide of P. sibiricum being steamed and shined for nine times and establish polysaccharide characteristic map of P. sibiricum by ancient processing and distillate. The effect of different enzymes on the extraction efficacy of P. sibiricum polysaccharides was also compared. Results The extracting effect of cellulase in single enzyme was the best. Moreover, cellulose and pectinase were also the most significant factor for extracting polysaccharides from P. sibiricum in four complex enzymes. The best extracting conditions for polysaccharide obtaining by RSM as follows:Cellulase temperature was 51℃, cellulase dosage was 2%, and the total polysaccharide content was (675.34 ±0.10) mg/g when the pH value of enzymolysis was 5.40. PMP-HPLC was used to detect four processed P. sibiricum, which contained mannose, ribose, rhamnose, glucose, and galactose. Glucose had a high proportion in P. odoratum, P. cyrtonema, and P. sibiricum polysaccharide, and P. filipes polysaccharide mainly composed of mannose. With the increase of processed times, the content of each monosaccharide increased first, then decreased, then increased and then tended to be stable. The content of mannose and galactose varied greatly. The content of monosaccharides in the distillate increased first and then stabilized, and the content changes of mannose and glucose were obvious. Conclusion The extraction efficiency and purity of P. sibiricum polysaccharides can be improved by single and compound enzymatic extraction with cellulase, and the extraction yield of polysaccharide in P. sibiricum could be affected by steamed and shined for nine times. PMP-HPLC characteristic map technology can be used to determine the content of the polysaccharide in processing of P. sibiricum accurately and reproducibly, which can be used for the quality evaluation of P. sibiricum and other Chinese herbal medicines.

湖北省食品药品监督管理局重点项目（20180103）；湖北省卫生健康委员会中医药面上项目（ZY2019M029）；湖北科技学院药学重点学科专项（2019-20YZ07）