Construction Of Fingerprint And Screening Of Antioxidant And Antibacterial Activities Of Hemerocallis Citrina Baroni

Hemerocallis citrina Baroni is a perennial herb belonging to the genus Hemerocallis of the Liliaceae Jussy.It is also known as Forgettable Grass,Golden Needle Flower and Anshencai.It is a traditional edible and medicinal plant in China.Flavonoids,phenolic acids,anthraquinones,dihydrofuran-γ-lactams,terpenoids and sterols are the main active ingredients in cauliflower.In addition,saponins,aliphatic and monocyclic derivatives have also been reported.Modern pharmacological studies have shown that Hemerocallis citrina Baroni has antidepressant,antioxidant,antibacterial,anticancer and insecticidal activities.Its main medicinal parts are flowers and roots,and stems and leaves are often used as waste in the field.At present,there is a lack of perfect quality evaluation system of Hemerocallis citrina Baroni,and the level of comprehensive resource utilization remains to be further improved.In previous studies,the antioxidant and antibacterial effects of Chinese flowering cabbage plants are more about the related experiments of crude extracts of Chinese flowering cabbage,and there are few further studies on the material basis of pharmacological activities of flower buds and roots of Hemerocallis citrina Baroni.In this paper,the HPLC and GC-MS fingerprints of 28 batches of sexual components from seven producing areas in China were established by using fingerprint technology,and the common peaks were confirmed.The differences in chemical components of different producing areas were analyzed.The antioxidant and antibacterial activities of different extracts from roots and buds of Hemerocallis citrina Baroni in Qidong area of Hunan Province were screened,and the chemical components were analyzed.Subsequently,the main active components were separated and identified by column chromatography and high performance preparative liquid chromatography.The above results can provide scientific reference for the identification of common compounds and the improvement of quality standards in different regions of China,and provide theoretical support for the material basis of antioxidant,antibacterial and other pharmacological activities.The results are as follows :1.The HPLC fingerprints of 28 batches of Hemerocallis citrina Baroni from different origins were established by HPLC analysis method combined with fingerprint technology.There were 8 common peaks in the fingerprints of each batch.By comparing with the chromatographic peaks of the reference substance,the chromatographic peaks 4,6,7 and 8were determined as chlorogenic acid,rutin,rhein and chrysophanol,respectively.The contents of two common peaks of rutin chlorogenic acid were determined.In the content determination,the RSD values of rutin and chlorogenic acid were 37.69 % and 4.67 %,respectively,indicating that different origins had great influence on the content of rutin,while chlorogenic acid was relatively stable.The classification results of cluster analysis are closely related to the actual origin of Hemerocallis citrina Baroni.2.The GC-MS fingerprint of Hemerocallis citrina Baroni from different habitats was established from the perspective of volatile substances.Combined with the fingerprint software of traditional Chinese medicine,14 common peaks were established,which were 3-furan methanol(peak 1),2-pentylfuran(peak 2),3-FURYLMETHYL ACETATE(peak 3),3.5-Octadien-2-ol(peak 4),trans-2-octene-1-ol(peak 5),nonanal(peak 6),1-nonanol(peak 7),decanal(peak 8),spirodicyclohexane(peak 9),(Z)-N-Butyl-2-buten-1-amine(peak 10),pentadecane(peak 11),dihydroactinidiolide(peak 12),trans-nerolidol(peak 13),alphaasarone(peak 14).The RSD values of the relative retention time of the common characteristic peaks were all 0.12 % and below,while the peak areas of the common peaks in each batch were quite different.In addition,the similarity is above 0.994,which reflects the stability of its quality to a certain extent.The relative content of common peaks was obtained by area normalization method.The clustering results showed that the volatile components from different habitats had little effect on the volatile components.3.FRAP method and DPPH method were used to evaluate the antioxidant activity of different extracts of flowers and roots.Microdilution method was used to evaluate the antibacterial activity of different extracts of flowers and roots,and the chemical constituents of the active parts were analyzed by ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry combined with MS and MS/MS data.The results showed that the total antioxidant capacity of ethyl acetate extract(6.88 mmol/g)was stronger than that of Trolox(6.59 mmol/g),and no obvious FRAP total antioxidant capacity was observed.The DPPH results showed that when the concentration of sample solution was0.4～1 mg/m L,the DPPH free radical scavenging capacity of the roots and ethyl acetate extracts of Hemerocallis citrina Baroni was significantly stronger than that of PG,which was not different from that of VC,and the scavenging rates were all greater than 90.8 %.After preliminary screening,it was found that the petroleum ether and dichloromethane extracts from roots and the petroleum ether and ethyl acetate extracts from flowers showed certain antibacterial activities,but the bactericidal effect was not obvious.The above parts were analyzed by UPLC-Q-TOF-MS combined with mass spectrometry,and 39 chemical components were preliminarily identified.4.Column chromatography was mainly used to separate the monomer compounds from the active extraction parts of the roots of Hemerocallis citrina Baroni.The structures were determined by high-resolution mass spectrometry and nuclear magnetic resonance to explore the potential active compounds.Finally,19 monomeric compounds were identified as bis(2-ethylbutyl)pathalate,bis(2-ethylhexyl)benzene-1,2-dicarboxylate,dihydroflavonoid glycosides,1,8-dihydroxy-3-methyl-anthraquinone,1,8-dihydroxy-3-carboxylanthraquinone,2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-tryptophan-4-one,capsicum oleoresin,cassiaquinone,quercetin-3-O-β-D-galactopyranoside,1.8-Dihydroxy-3-hydroxymethylanthraquinone,uracil nucleoside,Kaempferol,kaempferol-3,7-O-Ldirhamnoside,9-β-D-furanosyladenine,5-caffeoylquinic acid,Zeaxanthin,Quercitrin,Isoquercitrin,puerarin flavonoids.Compounds 1,2,7 were first found in Hemerocallis citrina Baroni plants.