高温瞬时灭菌对开心散有效成分含量及微生物学指标的影响

Effect of High-temperature Instantaneous Sterilization on the Content of Active Components and Microbiological Indexes of Kaixin San

目的:比较不同高温瞬时灭菌条件下开心散的灭菌效果,结合灭菌后样品中有效成分含量的变化,确定最佳灭菌条件。方法:采用高温瞬时灭菌法对2~8目的远志、人参、茯苓和石菖蒲4味中药的粗颗粒进行灭菌,分别考察160、165、170℃3个温度水平,灭菌时间分别为5、7、9 s。在洁净区对灭菌粗颗粒分别进行粉碎,过六号筛后均匀混合,制得开心散。参照《中华人民共和国药典》2020年版进行微生物限度检测,并采用高效液相色谱法对开心散灭菌前后的样品进行含量检测。结果:9个灭菌条件下开心散样品的需氧菌总数均小于1×104CFU·g^(–1)、霉菌和酵母菌总数均小于1×102CFU·g^(–1)。同时,控制菌项下的大肠埃希菌、沙门菌、耐胆盐革兰阴性菌均未检出,所有开心散灭菌样品均符合口服散剂的微生物限度要求。开心散中8个有效成分(远志[口山]酮Ⅲ、3,6′-二芥子酰基蔗糖、细叶远志皂苷、人参皂苷Rg1、人参皂苷Rb1、茯苓酸、β-细辛醚和α-细辛醚)的含量受不同灭菌温度和时间影响。结论:结合需氧菌、霉菌和酵母菌、控制菌结果和有效成分含量变化情况,最终确定灭菌温度160℃、灭菌时间5 s为开心散最佳高温瞬时灭菌条件。

Objective:The optimal sterilization conditions for Kaixin San were determined by comparing the effects of different high-temperature instantaneous sterilization conditions and combining with the change of index components in the samples after sterilization.Methods:The crude particles of 2-8 samples of Polygalae Radix,Ginseng Radix et Rhizoma,Poria cocos,and Acori Tatarinowii Rhizoma were sterilized by high-temperature instantaneous sterilization at 160,165 and 170℃for 5,7 and 9 s,respectively.Then,the sterilized crude particles were crushed respectively in the clean area,passed through a No.6 sieve and uniformly mixed to prepare Kaixin San.According to the Chinese Pharmacopoeia(2020 edition),the microbial limit test was carried out,and the contents of the samples before and after sterilization of Kaixin San were detected by high-performance liquid chromatography(HPLC).Results:The total numbers of aerobic bacteria of Kaixin San samples under 9 sterilization conditions were all less than 1×10^(4) CFU·g^(–1),and the total numbers of mold and yeast were both less than 1×10^(2) CFU·g^(–1).Meanwhile,no control bacteria,such as Escherichia coli,Salmonella,and bile salt-resistant Gram-negative bacteria,were detected,indicating that all sterilized Kaixin San samples met the microbial limit standards of oral dispersible powder as specified in the Chinese Pharmacopoeia(2020 edition).The contents of 8 index components(polygalaxanthoneⅢ,3,6ʹ-disinapoylsucrose,tenuifoliside,ginsenoside Rg1,ginsenoside Rb1,pachymic acid,β-asarone andα-asarone)in Kaixin San were affected by different sterilization temperatures and durations.Conclusion:According to the microbial testing results and the content changes in the index components,the optimal high-temperature instantaneous sterilization conditions for Kaixin San were determined to be a temperature of 160°C and a time of 5 s.