| Fritillariae cirrhosae bulbus is one of the most representative and valuable medicinal plants from Sichuan province.As a high-quality and effective traditional Chinese medicine for relieving cough and resolving phlegm,it has a long history of clinical application and a broad potential for developing the whole industry chain.However,the current development of Fritillariae cirrhosae bulbus industry is constrained by some bottleneck problems,such as insufficient resource supply,weak quality evaluation and control,lack of technical standards,and shortage of end products.To solve the problem of weak quality evaluation and control,improve the quality evaluation standards,and address the lack of end products,this research innovates on the basis of the existing quality evaluation of Fritillariae cirrhosae bulbus,and focuses on the following aspects.1.A method for simultaneous quantitative analysis of six index components in Fritillariae cirrhosae bulbus from different sources based on high performance liquid chromatography-evaporative light scattering detector(HPLC-ELSD)technique was established,and the quality differences of Fritillariae cirrhosae bulbus from different sources were compared and analyzed from the perspective of multi-component chemical evaluation.In this study,the contents of six index chemical components(Sipeimine-3β-D-glucoside,Sipeimine,Peimisine,Peimine,Peiminine,Hupehenine)in six species of Fritillariae cirrhosae bulbus were determined by HPLC-ELSD method.The methodological investigation results showed that the linear relationships of the six chemical components were good,and the correlation coefficients r were all greater than0.99.The precision results showed that the RSD values of the retention time and peak area of the six chemical components were all less than 2.0%.The repeatability results showed that the RSD values of the retention time and peak area of the six chemical components were all less than 2.5%.The stability results showed that the RSD values of the retention time and peak area of the six chemical components were all less than2.5%.The recovery rates of the six alkaloids ranged from 97.89% to 101.75%,which met the evaluation criteria for methodological validation.The content of chemical components in Fritillaria unibracteata Hsiao et K.C.Hsiavar.wabuensis(S.Y.Tang et S.C.Yue)Z.D.Liu,S.Wang et S.C.Chen wabuensis was the highest,significantly higher than other species,and the content of chemical components in Fritillaria unibracteata Hsiao et K.C.Hsia.was the lowest.Among the six chemical components,Sipeimine had the least detection frequency in 23 batches of samples,and Peimisine had the highest detection frequency and content.The comparison results of chemical components in different specifications showed that the content of chemical components in samples with smaller specifications was generally higher than that in samples with larger specifications.The comparison results of samples with different growth years showed that the content of chemical components in samples with 3-4 years of growth was higher.The fingerprint chromatograms of six species of Fritillariae cirrhosae bulbus samples were established,and the similarity of 23 batches of samples ranged from 0.385 to 0.918.The cluster analysis results showed that the content of six chemical components in samples from different origins of the same species was similar,and there were significant differences in the six chemical components and their contents among samples from different species.The principal component analysis results showed that the Fritillaria unibracteata Hsiao et K.C.Hsiavar.wabuensis(S.Y.Tang et S.C.Yue)Z.D.Liu,S.Wang et S.C.Chen and Fritillaria taipaiensis P.Y.Li had larger differentiation degree from other four species samples,and Fritillaria przewalskii Maxim.and the Fritillaria unibracteata Hsiao et K.C.Hsia samples had smaller differentiation degree and closer content of chemical components.2.An artificial sputum solution was developed for evaluating the phlegmification biological potency of Fritillariae cirrhosae bulbus,and a comprehensive quality evaluation method of Fritillariae cirrhosae bulbus based on the determination of phlegmification biological potency was established.The results of the stability study of the artificial sputum showed that the viscosity of the artificial sputum under natural conditions decreased by 0.11 m Pa·s after 120 min with RSD was 1.07%.The phlegmification potency at 120 min was used as the evaluation index,and the ranking of the efficacy of phlegm reduction was Fritillaria unibracteata Hsiao et K.C.Hsiavar.wabuensis(S.Y.Tang et S.C.Yue)Z.D.Liu,S.Wang et S.C.Chen,Fritillaria przewalskii Maxim.,Fritillaria unibracteata Hsiao et K.C.Hsia,Fritillaria taipaiensis P.Y.Li,Fritillaria cirrhosa D.Don,and Fritillaria delavayi Franch.Among them,Fritillaria unibracteata Hsiao et K.C.Hsiavar.wabuensis(S.Y.Tang et S.C.Yue)Z.D.Liu,S.Wang et S.C.Chen was the best and Fritillaria delavayi Franch.was the least effective in phlegmification.Analysis of the pattern of the phlegmification biological potency of the same origin samples: the comparative results of the phlegmification biological potency of all 6 samples showed that the phlegmification biological potency of the sample with smaller bulb size was lower,the phlegmification biological potency of the sample with medium size was the highest,and the septic potency of the sample with larger size increased proportionally with time.Except for Fritillaria przewalskii Maxim.,the increase of phlegmification biological potency of other basal samples was the greatest at 30～60 min,and the effect of phlegmification was the most obvious,indicating that the effective substances in Fritillariae cirrhosae bulbus could act on the artificial sputum faster in this time period.Comparison of the phlegmification potency of different origin samples with the same growth age: the phlegm chemotactic potency of the 5-year old samples was in the order of Fritillaria przewalskii Maxim.> Fritillaria unibracteata Hsiao et K.C.Hsiavar.wabuensis(S.Y.Tang et S.C.Yue)Z.D.Liu,S.Wang et S.C.Chen>Fritillaria unibracteata Hsiao et K.C.Hsia>Fritillaria taipaiensis P.Y.Li>Fritillaria cirrhosa D.Don> Fritillaria delavayi Franch.The results showed that the method can be used to evaluate the comprehensive quality of different Fritillariae cirrhosae bulbus samples,and there were large differences between the six basal samples of Fritillariae cirrhosae bulbus in terms of the effect of phlegm,and the quality evaluation and control study of the basal herbs of Fritillariae cirrhosae bulbus still needs to be strengthened.3.A composite solid beverage based on Fritillariae cirrhosae bulbus was developed,providing an application example for expanding the health products of Fritillariae cirrhosae bulbus.Fritillariae cirrhosae bulbus,Phyllanthi Fructus,and Platycodonis Radix were used as raw materials,and aspartame,citric acid,and maltodextrin were used as auxiliary materials to develop the composite solid beverage of Fritillaria cirrhosa.The particle size and dissolution rate of the raw materials were investigated,and 80-mesh fine powder was selected as the raw material.The orthogonal test was used to optimize the addition amount of the raw materials,and the optimal addition amount was 4g of Platycodonis Radix,2.5g of Phyllanthi Fructus,and 1.5g of Fritillariae cirrhosae bulbus.The single factor test and response surface design were used to optimize the optimal addition amount(based on the quality of the composite solid beverage of Fritillaria cirrhosa)of the sweetener aspartame,the sour agent citric acid,and the stabilizer maltodextrin,which were 0.93%,1.06%,and 5.3%,respectively.A product with light brown color,uniform tissue state,delicate taste,moderate sourness and sweetness,and certain throat-clearing and lung-moistening functions was obtained. |
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