Comparison Of Chemical Information Between Aconiti Kusnezoffii And Other Medicinal Parts And Study On Chemical Constituents Of Leaf Of Aconitum

Objective: Through a comprehensive study of the chemical components of dichloromethane and ethyl acetate extracts from the leaves of Aconitum Kusnezoffii,the chemical information of the leaves of Aconitum Kusnezoffii is identified,and the characteristic chemical components that cause differences in the leaves,flowers,and buds of Aconitum Kusnezoffii are screened,laying a preliminary research foundation for elucidating the pharmacological substance basis of Aconitum Kusnezoffii and its processing,quality evaluation,toxicity,and other studies.Methods: Take 2.3 kg of Aconitum Kusnezoffii leaves,first extract with 95%ethanol reflux method,then add an appropriate amount of water to the extract,stir to form a suspension,and sequentially extract with dichloromethane and ethyl acetate to obtain dichloromethane and ethyl acetate extracts.The dichloromethane and ethyl acetate extracts were repeatedly separated into a single compound using classical column chromatography,TLC,HL-20,and HPLC chromatography and chemical techniques.The one-dimensional and two-dimensional spectral data of a single compound were determined using a nuclear magnetic resonance instrument,and the structural formula of the single compound was determined by analyzing its spectral data.Determine the structural formula of a single compound by measuring and analyzing various spectral data.At the same time,a simultaneous determination method for the content of aconitine,hypoaconitine,and neoaconitine in the leaves of Aconitum Kusnezoffii has been established,providing a strong basis for the quality evaluation of this medicinal herb.On the basis of understanding the chemical components of the leaves of Aconitum Kusnezoffii,multivariate statistical analysis methods such as Principal Component Analysis(PCA),Orthogonal Partial Least Squares Discriminant Analysis(OPLS-DA),and Cluster Analysis(HCA)were used to compare the differences in chemical profiles of Aconitum Kusnezoffii and its leaves,flowers,and buds,and to screen the characteristic chemical components that caused the differences.Result: 22 compounds were isolated from different solvent extracts of Kusnezoffia leaves,namely stigmaster-4-en-3-one(1)and 6 β-Hydroxy-stigmaster-4-en-3-keto(2),5 α-Hydroxy-stigmaster-3-one(3),stigmaster-4-en-3-ol(4)β-Sitosterol(5),palmitic acid(6),oleic acid(7),linoleic acid(8)α-Linolenic acid(9),6,7-epoxy-α-Linolenic acid(10),obturatin(11),benzoic acid(12),p-hydroxybenzoic acid(13),apigenin(14),kaempferol(15),(10b S)8,9-dihydroxy-1,5,6,10 b tetrahydro-2H-pyrrolo [2,1a]-isoquinoline-3-one(16),aconitine(17),hypoaconitine(18),neoaconitine(19),carotene(20),kaempferol-3-O-β-D-xyloside(21),kaempferol-3-O-β-D-galactoside(22).Under experimental chromatographic conditions,the linear relationship(r>0.9995)between neoaconitine,hypoaconitine,and aconitine showed good performance over a wide concentration range.The average recovery rates of neoaconitine,hypoaconitine,and aconitine were97.6%,97.2%,and 97.2%,respectively,and the RSD was also less than 2.0%.The relative content changes of neoaconitine,hypoaconitine,and aconitine in different batches of Aconitum Kusnezoffii leaves were 0.393 ± 0.005～0.405 ± 0.008,0.244 ±0.003～0.251 ± 0.005,and 0.138 ± 0.001～0.143 ± 0.003 mg/g,respectively.The results of PCA and HCA analysis showed that the leaves and flowers of Aconitum Kusnezoffii were first clustered,and then sequentially clustered with Aconitum Kusnezoffii and Aconitum Kusnezoffii buds.The OPLS-DA load map analysis identified five main biomarkers that contribute to the classification of samples from different parts of Aconitum Kusnezoffii,including its leaves,flowers,and buds.The variables with significant contributions were screened using VIP values(with a VIP value>1 as the standard),which are the characteristic peaks corresponding to T6,T8,T2,and T1.The four components are the main landmark components that cause differences in components between Aconitum Kusnezoffii and its leaves,flowers,and buds.Conclusion: 22 compounds were isolated and identified from the ethanol extract of Aconitum multiflorum leaves.Through this experimental study,the basic information of Aconitum multiflorum leaves was clarified,solving the problem of unclear chemical composition.A simultaneous determination method for the content of aconitine,hypoaconitine,and neoaconitine in the leaves of Aconitum multiflorum was established through ultra-high performance liquid chromatography.This method is fast,simple,and stable,providing a strong basis for the quality evaluation of this medicinal material.Based on a clear understanding of the chemical components of the leaves of Aconitum multiflorum,the characteristic chemical components causing differences in Aconitum multiflorum and its leaves,flowers,and buds were analyzed using SIMCA multivariate data software.