| Trollius chinensis Bunge is a medicinal and edible plant,which has the functions of clearing heat and detoxifying,cooling blood and relieving sore throat,nourishing yin and clearing heat.Trollius chinensis plants are widely distributed in China and are easy to cultivate.At present,the research of Trollius chinensis mainly focuses on the chemical constituents of flavonoids,and lacks the research on alkaloids and phenolic acids.There is no comprehensive study on the quality evaluation and resource utilization of Trollius chinensis.In this paper,the extraction and purification of total alkaloids from Trollius chinensis were systematically studied by response surface method,and the content of total alkaloids from different habitats was determined to explore the antioxidant and antibacterial activities of total alkaloids from Trollius chinensis.The fingerprint was established by HPLC method to screen the characteristic peaks of Trollius chinensis,and the contents of orientin,vitexin and quercetin in Trollius chinensis were determined.The detection conditions and contents of vanillic acid and veratric acid were established and determined.Chemometrics was used to combine total alkaloids,phenolic acids and fingerprints to investigate the chemical diversity of Trollius chinensis in different producing areas from multiple perspectives,and three models of PCA,HCA and PLS-DA were established to classify different producing areas,in order to provide a theoretical basis for the subsequent identification of producing areas of Trollius chinensis.1.The content of total alkaloids in Trollius chinensis from 11 producing areas ranged from0.10～0.64 mg/g,with the highest content in Wutai Mountain of Shanxi and the lowest content in Greater Khingan Mountains of Inner Mongolia.The results showed that the best conditions for extraction and separation of total alkaloids were as follows,ethanol concentration 90%,liquid-solid ratio 39 m L/g,ultrasonic power 400 W,ultrasonic time 32 min,total alkaloids yield was 0.844 mg/g.Through in vitro antioxidant and bacteriostatic tests,it was found that the total alkaloids of Trollius chinensis had strong antioxidant capacity.Compared with DPPH and ABTS,the scavenging ability of superoxide anion was slightly weaker,and the inhibitory effect on Escherichia coli and Staphylococcus aureus was obvious.2.The content of orientin ranged from 8.558 to 23.098 mg/g,the content of vitexin ranged from 0.685 to 5.014 mg/g,the content of quercetin ranged from 0.901 to 2.980 mg/g,the content of veratric acid ranged from 0.219 to 1.192 mg/g,and the content of vanillic acid ranged from 0.284 to 0.834 mg/g.Among them,the content of orientin in Jiamusi,Hailun and Greater Khingan Mountains of Heilongjiang Province was higher(0.376～23.0989 mg/g,the average content was 21.622 mg/g),which was consistent with the conclusion of the existing papers and could be confirmed by each other.The content of vitexin in Genhe of Inner Mongolia and Ulagai grassland of Xilin Gol League of Inner Mongolia is higher,and the content of vitexin in Greater Khingan Mountains of Inner Mongolia and Aershan of Inner Mongolia is less.It is speculated that Inner Mongolia has a wide area and different climate effects.3.Chromatographic conditions: 254 nm for the detection wavelength;gradient elution was performed with 0.2% phosphoric acid(A)-acetonitrile(B),0～8 min,80%A;8～15 min,80%～75%A;15～20 min,75%～70%A;20～40 min,70%A;40～50 min,70%～80%A;50～60min,80%A.The flow rate was 1 m L/min,and the column temperature was 30°C.The response values of 11 batches of fingerprints and detected chemical components were obtained.The similarity judgment was performed using the 2012 version of the ’ Chinese medicine fingerprint similarity calculation evaluation system ’ software.The results showed that the similarity of73 % was above 0.85,and the quality was stable,but different producing areas could not be classified.4.The data of fingerprint,flavonoids,phenolic acids and total alkaloids were statistically analyzed by chemometrics.The 19 common peaks were reduced by PCA unsupervised principal component analysis,and 9 characteristic peaks including orientin,vitexin and quercetin were selected to establish PCA model.The characteristic peaks were combined with the data of vanillic acid,veratric acid and total alkaloids,and all the components were unified to establish the PCA model.According to the score plot,it was divided into three parts.The origin of Inner Mongolia and Heilongjiang was one category,and the origin of Wutai Mountain in Shanxi and Chengde in Hebei was one category.Bo Zhou and Aershan are the third category,and it is speculated that Bo Zhou suppliers purchase Trollius chinensis from Inner Mongolia.SIMCA analysis method was used to investigate the importance of several components,and 12 components were determined as quality markers to distinguish the origin of Trollius chinensis.The PCA model was verified by HCA clustering analysis and supervised PLS-DA model.The results showed that the PCA model was reliable and accurate.In this study,the optimal extraction and purification process of total alkaloids from Trollius chinensis was determined,and it was preliminarily determined that the total alkaloids from Trollius chinensis had antibacterial and antioxidant biological activities.In this paper,it was found that there were great differences in the chemical components of Trollius chinensis in different producing areas.Chemometrics was used to analyze the different types of chemical components in Trollius chinensis in different producing areas,and the quality markers that could distinguish the producing areas were screened out,and the classification model was established,which could preliminarily determine the producing areas of unknown samples.To provide reference for further rational application of Trollius chinensis and new drug research and development. |
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