Analysis Of Amides And Phenylpropanoids In Wolfberry And The Origin Identification Of Wolfberry

In this work,the fruit of Lycium barbarum as the research object,the research was carried out from the analysis and activity evaluation of the chemical constituents on the fruit of Lycium barbarum,and the origin identification of wolfberry.This work was divided into three chapters.The research contents and main results of each chapter are summarized as follows.Chapter 1.Literature studySection 1.Research progress on amides constituents of the Lycium genusIn this section,based on the systematic search and collation of the relevant literature,the composition types of amides in the Lycium genus were systematically summarized.The results showed that cinnamamides were the most abundant amides in the Lycium genus.It was also found that there were many glycosidated amides in the Lycium genus,which were mainly distributed in the fruit.The products of spermidine after glycosidylation are potential biomarkers of the Lycium genus.Section 2.Research progress of the bioactivity of amides in the Lycium genusIn this section,based on the systematic search and collation of the relevant literature,the biological activities of the amide components in the Lycium genus are summarized.The results showed that this kind of composition had anti-inflammatory,anti-oxidation,immunomodulatory,and other biological activities.At the same time,the prospect of its industrialization was prospected to provide reference for the efficient utilization of the Lycium genus.Chapter 2.Composition analysis and activity evaluation of amides and phenylpropanoids in wolfberrySection 1.Analysis of amides and phenylpropanoids in wolfberryIn order to clarify the types of resource chemical constituents in wolfberry.The chemical constituents of wolfberry were analyzed by UPLC-Q-TOF/MS.A total of 42 compounds were identified,including 32 hydroxycinnamamide alkaloids,7 phenylpropanoids,2 steroids,and 1 flavonoid by combining their retention time,mass spectrometry,and reference literature.At the same time,it was found that there are more isomers and unknown types of compounds in wolfberry.In order to clarify the fine structure of chemical constituents,the chemical constituents of wolfberry were isolated and identified by modern chromatography and spectral analysis.21 compounds were isolated and identified from the aqueous extract.Nine of them were alkaloids,including lycibarbarspermidine T(1),scotanamine D(2),lyciamarspermidine C(3),lycibarbarspermidine A(4),lycibarbarspermidine B(5),lycibarbarspermidine D(6),5-hydroxymethyl-pyrrole-2-carbaldehyde(7),4-[formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl]butanoic acid(8)and 2-(5-Hydroxymethyl-2-formylpyrrol-1-yl)propionic acid lactone(9);Ten of them were phenylpropanoids,including chlorogenic acid(10),trans-p-coumaric acid 4-O-β-D-glucopyranoside(11),cis-p-coumaric acid-4-O-β-D-glucopyranoside(12),trans-p-coumaric acid(13),cis-p-coumaric acid(14),methyl 4-O-β-D-glucopyranosyl caffeate(15),caffeic acid(16),lycibarbarcoumarin A(17),lycibarbarphenylpropanoid C(18)and lycibarbarphenylpropanoid D(19);Two other compounds were rutin(20)and 5-hydroxymethylfurfural(21).Compound 1 was a new compound,and among the known compounds,compounds 2,3,9,11,and 12 are reported from L.barbarum for the first time.Section 2.Bioactivity evaluation of alkaloids in wolfberryThe anti-inflammatory and neurotrophic effects of alkaloids were evaluated by the mouse macrophage(RAW 264.7)model and rat glioma cell(C6)model.The results based on RAW 264.7 cell model showed that alkaloids had no effect on cell proliferation at the concentration of 3-30 μM.Except compound 5,the other compounds could inhibit the NO release level of cells stimulated by LPS at the concentration of 3-30 μM.Based on the structure-activity relationship analysis of compounds 2-6,the activities of spermidine alkaloids are mainly related to the hydroxyl groups at C-3 and C-4 positions and the double bonds at C-7 and C-8 positions of caffeic acid derivatives.Based on the C6 cell model,compounds 1-3 and 9 could promote the expression of NGF,BDNF,and GDNF mRNA;Compound 4 could only promote the expression of NGF mRNA.Compound 5 could promote the expression of BDNF and GDNF mRNA.Compounds 6-8 could promote the mRNA expression of NGF and BDNF.The results provided a scientific basis for the further development of wolfberry.Chapter 3.Origin identification and quality analysis of wolfberry based on flavor Section 1.Research on origin identification based on electronic tongue technologyIn order to find a method to quickly identify the origin of wolfberry,this section studies the taste of wolfberry from different origins based on oral taste and electronic tongue technology.The results showed that there was a certain correlation between taste and electronic tongue,which proved that electronic tongue technology could objectively reflect the taste of wolfberry.Through the taste information radar map of the electronic tongue,we can directly see that the taste differences of wolfberry from different regions are mainly reflected in the salty and sour taste.Represents the response value of CTS sensor for saltiness:QH(1022.18±50.36)>NX(992.93±23.36)>GS(825.74±82.85)>IM(503.12±57.67);Represents the response value of AHS sensor for sour taste:IM(3633.61±80.62)>GS(3376.56±107.08)>QH(3323.69±175.72)>NX(3198.43±80.13);Represents the response value of NMS sensor for freshness:NX(1932.25±46.54)>QH(1889.26±54.17)>GS(1858.77±38.76)>IM(1842.07±40.21);Represents the response value of SCS sensor for the bitter taste:IM(841.10±0.68)>GS(815.70±1.75)>NX(792.89±0.73)>QH(790.42±1.83);Represents the response value of ANS sensor for sweet:QH(2115.96±76.91)>NX(2112.41±43.01)>GS(2023.29±50.26)>IM(1979.92±40.14).The multivariate statistical analysis methods were used to study the response values of each sensor.The results showed that the technology of electronic tongue could better distinguish the producing areas of wolfberry,and the main influencing factors were salt and sour taste.Based on the results,the Fisher discriminant model of the wolfberry producing area was established.The parameters of the model were in accordance with the statistical requirements,which indicated that the model was stable and reliable.This study provides a basis for the rapid identification of the origin of wolfberry.Section 2.Correlation analysis of flavor and quality of wolfberryIn order to clarify the relationship between the taste of wolfberry and its quality.UPLC-Q/TOF MS was used in this section to analyze and study samples from different producing areas,in order to verify the results of the electronic tongue.The results showed that LC-MS could distinguish the origin of wolfberry,and the differences were mainly in triterpenes,phenolamiddes,and organic acids.Based on the research results of functional substances on wolfberry,multivariate statistical analysis was conducted on the water-soluble chemical constituents of wolfberry from different producing areas.The results showed that there were differences in the chemical composition among samples from different producing areas,mainly reflected in glucose,fructose,acid polysaccharide,hydroxyproline(Hpro),proline(Pro),y-aminobutyric acid and other components.The correlation between the content of 43 water-soluble chemical components and the taste of wolfberry was analyzed.The results showed that most of the chemical composition and the taste have correlation.The differences in chemical composition of wolfberry could be reflected by taste level to some extent,and the results of the electronic tongue was verified to a certain extent.To elucidate the complex relationship between the flavor and chemical composition of wolfberry and to provide the basis for evaluating its quality by the taste of wolfberry in the later stageIn summary,the chemical constituents of the water extract of wolfberry were analyzed by UPLC-Q-TOF/MS.43 chemical components were identified,and a large number of nitrogen-containing compounds were found in water extract of wolfberry.Twenty-one compounds were isolated and identified from the water extract of Lycium barbarum by modern methods of separation and analysis.One of them was a new compound,and five known compounds were isolated from Lycium barbarum for the first time.The anti-inflammatory activities of the nitrogen containing compounds were evaluated.It was found that all the compounds except compound 5 could decrease the release of NO in LPS-stimulated cells at the concentration of 3-30 μM.The neurotrophic effects of nitrogen-containing compounds were evaluated.It was found that nitrogen-containing compounds could achieve better neurotrophic effects by increasing different types of neurotrophic factors.Based on the electronic tongue technology,a new method which can quickly identify the origin of wolfberry was established and verified by modern analysis methods.