ABSTRACT
OBJECTIVE@#Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China.@*METHODS@#In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas.@*RESULTS@#The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value < 0.05 associated with the separation of different origins were identified.@*CONCLUSION@#HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.
ABSTRACT
Objective: To provide the DNA molecular marker for the identification of Pseudostellaria heterophylla from the different idioplasms by analysis of rDNA ITS sequences. Methods: PCR amplification, cloning, and sequencing were carried out using specified primer, and the rDNA ITS base sequences were compared. Results: The ITS mutation extension was 623-624 pb among nine idioplasms of P. heterophylla. Thereinto, the ITS-1 was 224 pb and its G + C content was 52.91%-54.26%, the 5.8S rDNA was 155 bp and its G + C content was 54.49%-55.13%, the ITS-2 was 244-245 bp and its G + C content was 55.55%-56.41%. There were 17 mutation sites (2.72%) in the whole ITS sequences. There were 7, 7, and 3 mutation sites in ITS1, ITS2, and 5.8S, respectively. The different idioplasms had a number of specific single nucleotide mutation sites. Their homologies with each other were upwards 99.9% and their sequence genetic distances were 0.003-0.013. These results showed that the mutation in species from different producing areas and idioplasms was within no more than one species. Conclusion: The mutation of ITS sequences could be used to authenticate P. heterophylla from different idioplasms.
ABSTRACT
Objective To study rDNA ITS sequence similarities and differences between the de-virus seedlings and outside seedlings of Pseudostellaria heterophylla. Methods The sequences of ITS region (including ITS1, ITS2, and 5.8S) from nine de-virus seedlings and four outside seedlings were studied by the method of DNA sequence analysis. Results There were the same sequences in 5.8S region between the de-virus seedlings and outside seedlings of P. heterophylla. Variable sites lay in the initial position of ITS1 and termination of ITS2 basically. The de-virus seedlings from Liyang (Jiangsu Province) and Xuancheng (Anhui Province) had more variable sites. Phylogenetic dendrogram based on ITS was constructed by using UPGMA methods, which showed the inherent relation at the level of molecular biology. Conclusion The analysis of rDNA ITS of P. heterophylla has further clarified the inherit stability of de-virus seedlings at the molecular level.