ZHOU Yong-yi,XUE Jia,CHEN Hai-jie,et al.Analysis and Evaluation of Multiple Chemical Constituents in Different Parts of Lysimachiae Herba[J].Journal of Instrumental Analysis,2023,42(08):930-942.
ZHOU Yong-yi,XUE Jia,CHEN Hai-jie,et al.Analysis and Evaluation of Multiple Chemical Constituents in Different Parts of Lysimachiae Herba[J].Journal of Instrumental Analysis,2023,42(08):930-942. DOI: 10.19969/j.fxcsxb.23040102.
Analysis and Evaluation of Multiple Chemical Constituents in Different Parts of Lysimachiae Herba
A method based on ultra high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry(UHPLC-Triple Quad-MS/MS) was developed for the simultaneous determination of flavonoids,organic acids,nucleosides and amino acids in the stems,leaves,flowers and fruits of Lysimachiae Herba,and multivariate statistical analysis was used to compare the different parts of Lysimachiae Herba.The preparation conditions were optimized by single factor test.The samples were separated on an Agilent Extend-C,18,(2.1 mm × 100 mm,1.8 μm) analytical column at 30 ℃ by gradient elution,with 0.1% formic acid-methanol as mobile phase at a flow rate of 0.30 mL/min,and analyzed under multiple-reaction monitoring(MRM) mode.Based on the results of content,the different parts of Lysimachiae Herba were comprehensively evaluated by the combination of hierarchical cluster analysis(HCA),principal component analysis(PCA),partial least squares discriminant analysis(PLS-DA),one-way analysis of variance(One-way ANOVA) and grey relation analysis(GRA).The optimal extraction conditions were using 80% methanol as the extraction solvent,with solid-liquid ratio of 1∶10,and ultrasonic extraction of 60 min at room temperature. The results showed that the peaks for 40 constituents were sucessfully separated,with their correlation coefficients(,r,2,) not less than 0.999 2.The limits of detection(LODs) and limits of quantitation(LOQs) were in the range of 0.21-19.76 ng/mL and 0.71-65.88 ng/mL,respectively.The average recoveries ranged from 99.1% to 102%,with RSDs not more than 2.9%.HCA and PCA could be used to discriminate samples of different parts,and gather stems,leaves,flowers,and fruits into one category,respectively,indicating that the same samples from different batches had stable quality.As could be seen from the heat map,there were fewer constituents with high content in the stem,and more constituents with high content in the leaves and flowers.Under the condition that VIP was greater than 1,PLS-DA screened a total of 13 important constituents leading to the difference in content.The results of one-way ANOVA showed that there were significant differences in the content of flavonoids,organic acids,nucleosides,and amino acids in different parts.According to the column stacking diagram of four types of constituents,the total content in leaves and flowers was higher,while the total content in stems was the lowest.However,it was difficult to directly determine the comprehensive quality of stems,leaves,flowers,and fruits.Therefore,GRA was used to evaluate the four parts,and the results showed that the comprehensive quality of flowers was the best,followed by leaves,and the comprehensive quality of stems was the worst.In conclusion,the results of multivariate statistical analysis showed that the content of 40 multiple active constituents in different parts of Lysimachiae Herba varied significantly,among which the comprehensive quality of flowers and leaves was better.The simultaneous determination of multiple chemical constituents combined with multivariate statistical analysis is accurate and reliable,and it could be used for the comprehensive evaluation on the quality of Lysimachiae Herba,which provides a scientific basis for the rational harvesting and comprehensive development and utilization of Lysimachiae Herba.
Lysimachiae Herbadifferent partsultra high performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry(UHPLC-Triple Quad-MS/MS)multiple chemical constituentsmultivariate statistical analysis
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