The Study On Erthworm Extraction Process Optimization And Rapid Quality Analysis Of Key Production Links By Near Infrared Spectroscopy

As a unique resource in China,traditional Chinese medicine(TCM)plays an important role in the field of medical and health care.However,the composition of TCM is complex and the quality is uneven.The lack of process control methods in key production process restricts the process of TCM internationalization.Using new methods and techniques of modern science,with the integration of multi-disciplinary technology to develop TCM is necessary.Taking the related components of efficacy as the quality index,improving the dissolution of active ingredients is an important prerequisite to ensure the safety and effectiveness of TCM.The extraction process is the main production in the production process of most TCM,and is also one of the key production links.The content of active ingredients in different extraction processes varies greatly,resulting in differences in the quality and efficacy of the products.Through modern analysis technology,and applying process analysis technology(PAT)into the production process can help to establish timely and effective process control analysis method,so that we can improve production efficiency and achieve quality controllable of production process to reduce the batches difference,thereby,we can ensure quality uniform of product.Near infrared spectroscopy(NIRS)has become a common method in PAT because of its characteristics of fast,non-destructive,low cost and green pollution-free.At present,it has been widely used in the field of TCM origin discrimination,authenticity discrimination and quantitative analysis.Qilong capsule is mainly used for the treatment of cardiovascular diseases.Earthworm is one of its main medicinal herbs and it was researched in this study.According to the existing extraction process,combined with the therapeutic effect of drugs,the index components was choosed and the extraction process of earthworm was optimized based on design of experiment(DoE).Furthermore,NIRS was introduced to establish the rapid evaluation method of earthworm and extraction process quality combined with chemometrics method,which provides reference for improving product quality and ensuring product consistency.The specific research includes the following three aspects:(1)Study on the establishment of rapid model of geographical origins discriminate and main components content in earthworm.Antaris II Fourier transform near-infrared spectroscopy was used to discriminate the earthworm from five producing areas in Weifang,Liangshan,Shandong,Tangshan,Hebei,Ninghe and Baodi,Tianjin.The partial least squares-discriminate analysis(PLS-DA)model was established through spectral pretreatment and modeling band selection,and the recognition rate and rejection rate of the model were 100%,which confirmed the feasibility of using NIRS to identify the different geographical origins of earthworm.Based on the partial least squares(PLS)algorithm,the quantitative analysis models of guanosine and inosine in earthworm were established.The optimized parameters of guanosine quantitative model,correlation coefficient of calibration(RC),correlation coefficient of predication(RP),root mean square error of calibration,(RMSEC)and root mean square error of predication(RMSEP)were 0.9752,0.9439,0.06185 mg/g,0.07752 mg/g,respectively.The parameters of inosine quantitative model,RC,RP,RMSEC and RMSEP were 0.9550,0.9272,0.04547 mg/g and 0.05354 mg/g,respectively.The model has high prediction accuracy,which proves that NIRS can be used for the rapid determination of the guanosine and inosine content in earthworm.This study provides a rapid evaluation method of the quality of earthworm,and can be used to guarantee the quality of the product from the source.(2)Optimization of earthworm extraction process based on DoE.In this study,the extraction process of earthworm was optimized by the idea of DoE,and the key factors of solid-liquid ratio,time and temperature were selected.The content of soluble protein and lumbrukinase potency was the evaluation index,and the solid-liquid ratio of 1:16,1:18,1:20,the extraction time of 16,18,20 h,the temperature of 35,40,45℃ were selected.The orthogonal test was carried out according to the L9(34)orthogonal table.The results of orthogonal experiment were analyzed through visual analysis and variance analysis.Combined with the conservation of resources and energy,the optimal extraction process was determined,that is,the earthworm was added 20 times purified water to extract for 16 hours at 35℃ To further verify the optimized process,the results showed that under the conditions of this process,the extraction rate of soluble protein and lumbrokinase in earthworm was higher and the contents fluctuation range was smaller,which proved that the selected process conditions were stable and reliable.This can be used for upgrading and optimizing the related products of earthworm.(3)Study on quality rapid analysis of earthworm extraction process by NIRSIn the process of extracting the earthworm,the offline extract sample was collected according to the change trend of the main component content.The spectra of extract samples were collected by Antaris II Fourier transform near-infrared spectroscopy,and the reference data of soluble protein,guanosine and inosine content were correlated with spectra by chemometrics method.The quantitative analysis model is established by the selection of the pretreatment method and modeling bands.Finally,the quantitative model of soluble protein was established by support vector machine regression(SVR),and the Rc,Rp,RMSEC and RMSEP values of the model were 0.9685,0.9716,0.0599 mg/mL and 0.08255 mg/mL,respectively.The quantitative model of guanosine and inosine was established by PLS,in which the RC,Rp,RMSEC and RMSEP values of the guanosine quantitative model were 0.9783,0.9711,0.01046 mg/mL,0.00975 mg/mL,,respectively,and the values of the inosine quantitative model were 0.9828,0.9894,0.00524 mg/mL,0.00328 mg/mL,respectively.The results show that NIRS can be introduced into the process of earthworm extraction.The quantitative model can be used to quickly determine the content of active components in the extraction process.Based on the control of existing physical parameters in the extraction process,combined with the control of active component parameters,real-time feedback in production process can be extracted,so the information can help to guide the production,which is conducive to ensuring the quality consistency of the extraction process of earthworm.