穿心莲饮片霉变前后的4种内酯类成分含量变化

Changes in the content of 4 kinds of lactones components in Andrographis herba decoction pieces before and after mold contamination

中文关键词:  穿心莲饮片  穿心莲内酯  霉菌污染  高效液相色谱法

英文关键词:Andrographis herba decoction pieces  andrographolide  mold contamination  high performance liquid chromatography

中文摘要:

      目的 研究穿心莲饮片霉变前后4种穿心莲内酯类成分含量变化, 分析不同霉变程度对穿心莲饮片4种内酯类成分含量的影响。方法 参照2020年版《中国药典》四部通则1108“中药饮片微生物限度检查: 微生物计数法”进行霉菌和酵母菌计数方法的建立和验证, 采用建立的方法对霉变前后的穿心莲饮片进行霉菌计数; 采用高效液相色谱法测定霉变前后穿心莲饮片中4种穿心莲内酯成分含量。结果 穿心莲饮片对方法适应性验证的黑曲霉和白色念珠菌抑制作用均较小, 采用样品(1:10)稀释度时有杂菌干扰影响检测, 当样品稀释到1:100时杂菌干扰减少, 2种试验菌回收率比值都在0.5~2.0范围内故可采用样品(1:100)或以上稀释级做霉菌及酵母菌计数并采用平皿倾注法进行检验。4种穿心莲内酯成分含量高效液相色谱法测定采用制定好的色谱条件下4种成分在相应浓度区间线性良好(r>0.9999), 回收率为95.31%~104.14%, 相对标准偏差小于5% (n=6); 20批次穿心莲饮片霉变后脱水穿心莲内酯含量平均减少最大, 为62%, 新穿心莲内酯含量平均减少其次, 为69%, 穿心莲内酯含量平均减少为75%, 去氧穿心莲内酯含量平均减少最少, 为88%。结论 在穿心莲饮片霉变后对去氧穿心莲内酯含量减少较少, 而对穿心莲内酯、新穿心莲内酯、脱水穿心莲内酯含量减少较大, 故要对穿心莲饮片的储存过程中要定期监控, 保证其免受微生物(霉菌)污染, 保证用药的安全性和有效性。同时, 通过研究霉变对穿心莲饮片有效活性成分含量的影响, 为建立更完善的穿心莲饮片质量标准提供依据。

英文摘要:

      Objective To investigate the changes in the content of 4 kinds of andrographolide components in Andrographis herba decoction pieces before and after mold contamination, and analyze the impacts of different mold contamination degrees on the content of these 4 kinds of lactones. Methods According to the general principle 1108 microbial limit test for traditional Chinese medicine decoction pieces: Microbial enumeration tests in volume IV of the Chinese pharmacopoeia (2020 Edition), the method for enumeration of molds and yeasts was established and validated. The established method was used to count molds in Andrographis herba decoction pieces before and after mold contamination. High performance liquid chromatography was employed to determine the content of 4 kinds of andrographolide components in Andrographis herba decoction pieces before and after mold growth. Results Andrographis herba decoction pieces showed weak inhibitory effects on Aspergillus niger and Candida albicans during the method suitability test. When the sample was diluted at 1:10, miscellaneous bacterial interference affected the detection. When the sample dilution reached 1:100, the interference was reduced, and the recovery ratios of the 2 kinds of test strains were both within the range of 0.5–2.0. Therefore, a sample dilution of 1:100 or higher could be used for the enumeration of molds and yeasts, with the plate pour plate method adopted for the test. In the high performance liquid chromatography determination of the 4 kinds of andrographolide components under the established chromatographic conditions, the 4 kinds of components exhibited good linearity in their respective concentration ranges (r>0.9999), with recoveries ranging from 95.31% to 104.14% and relative standard deviations less than 5% (n=6). After 20 batches of andrographolide slices were moulded, the content of dehydrated andrographolide was reduced by 62% on average, the content of new andrographolide was reduced by 69% on average, the content of andrographolide was reduced by 75% on average, and the content of deoxyandrographolide was reduced by 88% on average. Conclusion After mold contamination of Andrographis herba decoction pieces, the content of deoxyandrographolide decreases slightly, while the content of andrographolide, neoandrographolide and dehydroandrographolide decrease significantly. Therefore, regular monitoring during the storage of Andrographis herba decoction pieces is necessary to ensure they are free from microbial (mold) contamination and to guarantee the safety and effectiveness of medication. Meanwhile, this study on the impact of mold contamination on the content of active components in Andrographis herba decoction pieces provides a basis for establishing a more comprehensive quality standard for Andrographis herba decoction pieces.