Research On The Sterilization Technology Of Safflower And Gastrodi

Objective:The visualization analysis software was used to analyze the application status and trend of traditional Chinese medicine（TCM）sterilization methods in the past 20 years,and the actual sterilization unit operation was carried out with Carthamus and Gastrodiae powder as an example.The advantages,disadvantages and suitable range of different sterilization techniques applied to the TCM powder were discussed in depth,so as to provide reference for the selection of appropriate sterilization methods in the actual production and application of Carthamus and Gastrodiae powder in the later stage.Methods:1.Literature analysis:bibliometric methods and visual analysis software were used to conduct an overall analysis of the field of traditional Chinese medicine sterilization in recent years,and to explore the application status and trend of traditional Chinese medicine sterilization methods.2.Establishment of fingerprint conditions of CarthamusThe chromatogram of Carthamus was investigated from chromatographic column,processing method,mobile phase,wavelength and other chromatographic conditions,and finally the fingerprint conditions were determined.3.Comparison of different sterilization processes of Carthamus and gastrodia:（1）Sterilization:Carthamus and gastrodia powder wet heat sterilization,superheated steam instantaneous sterilization,⁶⁰Co-γ-ray irradiation sterilization and electron beam irradiation sterilization.（2）Trait comparison:the effects of four different sterilization methods on the traits of Carthamus tinctorius and Gastrodia were compared.（3）Comparison of bacterial content:the bacterial content of Carthamus tinctorius and gastrodia powder after sterilization by four different methods was compared.（4）Content determination:compare the influence of four different sterilization methods on the content of Carthamus tinctorius and gastrodia raw powder.4.Refinement study of irradiation sterilization（1）Comparison of characteristics,bacterial content,content and fingerprint of Carthamus and Gastrodia treated by different irradiation sterilization methods.（2）Comparison of traits,bacterial content,content and fingerprints of Carthamus tinctorius and Gastrodia under different irradiation doses.（3）Radiation-chemical pattern recognition analysis of safflower and gastrodia elata with different irradiation doses.Results:1.After screening,a total of 322 literatures were included in this study.Author cooperation network analysis found that the core research teams in the field of traditional Chinese medicine sterilization were mainly formed,such as China Institute for Food and Drug Control and Guangzhou Institute of Drug Control.From the comprehensive analysis of the number of publications,the authors’institutions and keywords,it was found that with the improvement of production requirements,irradiation sterilization has become a research hotspot in the field of traditional Chinese medicine sterilization due to its advantages of high efficiency,good effect and low cost.2.Establishment of safflower fingerprint conditions:The HPLC spectrum of safflower was investigated from the chromatographic column,processing method of sample,mobile phase,wavelength and other chromatographic conditions,and the final fingerprint conditions were determined as follows:chromatographic column:ALLtima C₁₈（4.6×250mm,5μm）,mobile phase:0.5%aqueous phosphoric acid solution（A）-acetonitrile（B）,gradient elution 0-5 min,95%-95%A;5-15 min,95%-92%A;15-22 min,92%-92%a;22-32 min,92%-87%A;32～40min,87%A;40～50min,87%～82%A;50～60min,82%A;60-65 min,82%-67%A;65-70 min,67%A;70-85min,67%-0%A;85-90 min,0%A,detection wavelength:280nm,treatment method:water,column temperature:25℃,injection volume:10μL,flow rate:1.0ml/min.3.Comparison of different sterilization processes of Carthamus and gastrodia:Damp heat sterilization,superheated steam instantaneous sterilization,⁶⁰Co-γ-ray irradiation sterilization and electron beam radiation sterilization were used to treat the native powder of gastrodiae and Carthamus,respectively.The effects of different sterilization methods on gastrodiae and Carthamus were compared.Damp-heat sterilization uses high-pressure saturated water steam or circulating steam sterilization,boiling water spray and other means to make the proteins and nucleic acids in the microbial cells denatured and the microorganisms die.The sterilization is complete,the method is simple,and the equipment is simple.However,high temperature and high humidity have changed the medicinal components and traits of Gastrodia and Carthamus,which have a great impact on the primary medicinal powder,and should not be used as the first choice of sterilization of primary medicinal powder.Superheated steam instantaneous sterilization takes a higher temperature to sterilate the material,which greatly shorts the sterilization time,but it still has an impact on the character and composition of Carthamus.Due to the short sterilization time and high sterilization temperature,Carthamus with a large initial bacterial content still has the problems of incomplete sterilization and large loss of composition content after superheated steam instantaneous sterilization.As a kind of"cold sterilization",irradiation sterilization is carried out under normal temperature and pressure,and has no significant effect on the characteristics of the primary medicinal powder of Gastrodia and Carthamus,so irradiation sterilization can be preferred as the sterilization method of the primary medicinal powder.4.Radiation sterilization process research:compare the characteristics,bacteria content,main component content and fingerprint of gastrodia and safflower powder with different radiation sources and different radiation doses.At the dose of 10k Gy,there was no significant difference in the characters of Carthamus tinctorii and Gastrodiae after ⁶⁰Co-γ-ray irradiation and electron beam irradiation sterilization.Under the condition of low dose（2k Gy）irradiation,the total number of bacteria could reach the national drug health standard（no more than 1000 cfu/g per 1g,no more than 100 CFU/g per 1 ml,no control bacteria could be detected）.After storage for 3 months,it was found that the higher the irradiation dose,the lower the total number of bacteria detected.The results showed that the increase of irradiation dose was beneficial to the storage of the raw powder of safflower and Gastrodia,and the sterilization effect of ⁶⁰Co-γ-ray irradiation was better than that of electron beam irradiation at the same dose.After 2,6,8,10k Gy irradiation and electron beam irradiation of the raw powder of safflower and Gastrodiae,at the dose of 10k Gy,the difference of electron beam irradiation of safflower,gastrodiae ⁶⁰Co-γirradiation and electron beam irradiation of gastrodiae within 10k Gy was not significant,but the composition of safflower changed to some degree after 10k Gy⁶⁰Co-γirradiation.Therefore,the ⁶⁰Co-γirradiation of safflower should be controlled within 0-8 k Gy.Conclusion:The previous research on the relevant literature in the field of traditional Chinese medicine sterilization in the past two decades through bibliometric methods showed that irradiation sterilization has become a research hotspot in the field of traditional Chinese medicine sterilization due to its advantages of high efficiency,good effect and low cost.Carthami flos and Gastrodiae gastrodiae were taken as the research subjects,and the treatments were damp heat sterilization,superheated steam instantaneous sterilization,60Co-γ-ray irradiation sterilization and electron beam irradiation sterilization,respectively.The experimental results showed that irradiation sterilization had little effect on the original medicinal powder of traditional Chinese medicine,and could be preferred as the sterilization method of the original medicinal powder.After determining the conditions of safflower fingerprint,the fingerprints of safflower and Gastrodia irradiated by different radiation sources and different radiation doses were further compared and analyzed.At the dose of 10k Gy,the changes of electron beam irradiation of safflower,⁶⁰Co-γirradiation of Gastrodia and electron beam irradiation of Gastrodia in 10k Gy were not significant.After irradiation with 10k Gy⁶⁰Co-γ,the composition of safflower changed to some extent.The ⁶⁰Co-γirradiation of safflower should be controlled within 0-8 k Gy.⁶⁰Co-γ-ray irradiation sterilization can be preferred for the sterilization process of the primary powder of safflower and Gastrodiae.