| Objective: The characteristics and microstructure of Codonopsis pilosula from different origin were identified and their differences were compared.The chemical constituents of 90% methanol extracts from Codonopsis pilosula from different bases were analyzed and identified.The components of free sugar extract from Codonopsis pilosula were analyzed and compared.To determine the sweetness of different Radix Codonopsis pilosula free sugar extraction liquid and use gas chromatography-mass spectrometry(GC-MS)technology as evidence,a new method for detecting the sweetness of Codonopsis was constructed and the differences of the sweetness of different Radix Codonopsis pilosula were compared.Method: Observe the shape,color,texture,section,etc.The length of the sample is measured with a tape measure,and the diameter of the sample is measured by vernier caliper.Determination of characteristics difference of Codonopsis pilosula from different base sources.The random sampling method to select samples from different sources,basal part of stem 1 cm at the cross,take 0.5 cm thick materials,fresh products directly with FAA fixation,dried with 10% glycerin solution after softening,FAA fixation.The routine method of paraffin serial sections,every 5 tablets a total of7,each batch of film thickness were 10~15 ?m,safranin green staining,neutral balata,fixed camera,determination of xylem,phloem and root radius(measured the number of n=10),catheter diameter,vessel wall the thickness(n=50),analyzed by SPSS 18 software in different source structure difference of Radix Codonopsis.UPLC-Q-TOF-MS/MS technology was used to identify the chemical constituents of90% methanol fraction of Codonopsis pilosula.The UPLC method was used with the condition that the column was Agilent Poroshell 120 EC C18(3.0 × 50 mm,2.7 mm)column;The detection wavelength was set at 254 nm;The mobile phase was eluted with gradient by mixture of methanol and water(0.1% formic acid);The column temperature was 25 ?;The flow rate was 0.7 m L/min;The injection was 10 ?L.Agilent Quadrupole-Time of Flight mass spectrometry instrument(1290 Series HPLC system);electrospray ion source(ESI);the positive and negative two ionization mode detection;the volume flow of dry gas was 6.8 L/min,the dry gas temperature was325 ?,the atomization pressure was 0.7 Bar.In the positive ion mode,the capillary voltage was 4.5 k V,the capillary voltage was 4.0 k V under the negative ion mode,the fragmentation voltage 130 V,and the scanning range of m/z 50~1500.A variety of oligosaccharide compounds were extracted from Codonopsis pilosula by shaking extraction.The composition and structure of the oligosaccharides in Codonopsis pilosula were analyzed by GC-MS after acetylation.The GC method was used with the condition that the column was VF-23ms(30 M 0.25 mm×0.25 mm)capillary column;The carrier gas was high purity helium;The column flow rate was 1.0m L/min;The split ratio was 45:1;The inlet temperature was 220?;The column pressure was 73.0 kpa;Heating up mode of GC by programmed temperature.The electron bombardment was ion(EI)source;The temperature of the ion source was220?;The interface temperature was 250?;The mass spectrum condition was mass scanning range m/z 45~550.Monosaccharide solution of Codonopsis pilosula was prepared by extraction of total sugar from pure water and precipitation with 80%ethanol.The sweetness of 30 batches of three Codonopsis pilosula samples from different origins was detected by WYT-J Hand-held Sugar Meter and their differences were compared.The composition and structure of the oligosaccharides in Codonopsis pilosula were analyzed by GC-MS after acetylation.The GC method was used with the condition that the column was VF-23ms(30 M 0.25 mm×0.25 mm)capillary column;The carrier gas was high purity helium;The column flow rate was 1.0m L/min;The split ratio was 45:1;The inlet temperature was 220?;The column pressure was 73.0 kpa;Heating up mode of GC by programmed temperature.The electron bombardment was ion(EI)source;The temperature of the ion source was220?;The interface temperature was 250?;The mass spectrum condition was mass scanning range m/z 45~550.Result: Cross section of different origin and age of Radix Codonopsis mainly in cork cell columns,stone cells,phloem outer fissure,phloem and xylem accounted for the proportion of root cross section.This method was used to identify most of the compounds of the 90% methanol extracts from three different origins of Radix Codonopsis pilosula.A total of 38 compounds were identified by the identification of two fragment ions.Among them,all the three Radix Codonopsis pilosula all contain26 common chemical constituents including phenylpropanoside,alkaloids,amino acids,etc.,and there are different chemical components in different Radix Codonopsis pilosula.The optimized method was used to analyze 24 batches of three samples of Codonopsis pilosula from different habitats,and to identify seven kinds of free sugar components including sorbose,galactose,glucose,D-mannose,fructose,inositol,sucrose and so on in three Codonopsis pilosula.The optimized method was used to analyze the samples.Conclusion: It is found that the structure of the Radix Codonopsis pilosula is different and provides a reference for the identification of different Radix Codonopsis pilosula.UPLC-Q-TOF-MS/MS can be used in the qualitative analysis of the components of the Radix Codonopsis pilosula in the aspects of retention time,UV absorption spectrum,precise relative molecular mass,molecular formula and structural debris.A rapid and accurate analysis method for chemical constituents of Codonopsis pilosula was compared.GC-MS technology can be used for qualitative and quantitative analysis of the free sugar components of Codonopsis pilosula by retention time,molecular formula and structural fragments.A fast and accurate method for finding the free sugar components in Codonopsis pilosula was provided.The extraction conditions of free sugar of Codonopsis pilosula were optimized,the objective index was linked with the sweetness,and an objective and reliable evaluation method was provided for the sweetness detection of Codonopsis pilosula.The objectivity of the sweetness measured by the content of free sugar and the content of sugar was tested by GC-MS. |
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