| The complex system means that the analysis system contains a large number of known or unknown non-target components that interfere with the detection of the target component.However,the qualitative and quantitative analysis of multi-objectives in complex systems has always been a hot and difficult point in the development of analytical techniques.Traditional Chinese medicine is a typical complex system.A sample contains various substances with different structures and different physical and chemical properties.The stability and ionization degree of all kinds of substances are different,so the corresponding optimal parameter settings will also have The difference.A single detection condition does not completely express the structural information of the compound:due to factors such as mobile phase conditions,impurity interference,and low target content,some compounds are difficult to detect or cannot generate multi-level fragmentation data.If the quantitative test for a specific component can be automatically optimized by the instrument in order to obtain the best results,but the experimental objects of the complex system are generally non-directional compounds,structural information is incomplete and lack of standard substance control,it is difficult to carry out targeted optimization.There is a need for an optimization method that can be interposed between the two,which can increase the detection sensitivity of the target class compound without requiring a large amount of standard product and method development time.Therefore,it can improve the detection efficiency of complex systems and contribute to the modernization of traditional Chinese medicine.ObjectiveBy optimizing the parameters of the instrument,the accuracy and efficiency of the generation of first-order and multi-level ion data in the complex system of high isoflavone and steroidal saponins in Ophiopogon japonica are enhanced,and a data quality optimization strategy for the above two types of components is constructed.And based on this as a basis to study its cracking law and metabolites,build a technical method to enhance the overall analysis efficiency.Methods1.The mass spectral response and peak areas of the three standards in the positive and negative ion modes under different mass spectrometric detection parameters were investigated by means of isocratic elution at the same concentration,and a detection strategy for specific components of Ophiopogon japonicus was developed based on the trend..At the same time,the cracking behaviors of the three standards under different energy levels in different cracking modes were investigated and optimized and summarized.2.UPLC-Orbitrap-HCD method was used to identify the structure of high isoflavones in Ophiopogon japonicus extracts using the mass spectrometry parameters and cleavage energy optimized in the previous experiment.3.UPLC-Orbitrap-CID method was used to analyze and identify the in vivo metabolites of methylindigoflavanone A,Ophiopogon japonicus D,and ruscogenin using the optimized mass spectrometry parameters and fragmentation energy in the previous experiment.And infer the metabolic pathway.Results1.According to the peak area and change trend of each compound,the effect of capillary temperature on the stability of ions was determined to determine the optimum detection conditions for various components:5kV ionization voltage in positive ion mode and-4kV in negative ion mode.Ionization voltage;high capillary temperature of 350℃ was used to detect high isoflavones and Ophiopogon japonicus(negative ions),and a lower temperature of 300℃was used for detection when detecting ruscogenin.The cracking energy is referenced to the signal intensity and the type of debris at each energy level.To ensure that the high isoflavone components of Ophiopogon japonicae have better lysis results,the HCD model was used to detect 120/150 collision energy.Rusocinogen can use CID and HCD,and the cleavage energy is 35 units.2.According to the obtained accurate molecular weight data,retention time and feature fragment ion screening,68 high isoflavone components were identified in the total extract of Ophiopogon japonicus,of which 26 were high isoflavones and 31 were high isoflavones.There are 11 flavonoid glycosides.3.Based on the obtained accurate molecular weight data,retention time,and characteristic fragment ion screening,a total of 11 metabolites of methylidene high homoisoflavone A were detected in rat biological samples,9 of which were in the blood,3 In the faeces,the reactions involved are glucose and glucuronidation,sulphation and aldehyde formation.A total of 44 metabolites were detected in a rat biological sample,including 14 in the blood,22 in the urine,and 23 in the feces.The reactions that occur in the body are mainly hydroxylation,dehydrogenation,glucuronide binding and the like.Conclusion1.This topic has carried on the preliminary exploration to the influence of the mass spectrometry detection parameter on the compound,has determined the optimized detection parameter and the cracking energy level for the Ophiopogon flavonoids component,the Ophiopogon saponin composition,has laid the foundation for the further research later.2.Based on the optimized conditions,the structure of the high isoflavones was identified,and the metabolites of the two monomers were identified,which proved that the method can achieve the expected goals,and it is also the study of the material basis of Ophiopogon japonicus.Provides some basic scientific evidence. |
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