Abstract
The aim of this study was to explore the possible antibacterial components of Salvia miltiorrhizae on Pseudomonas aeruginosa using a combination of chemical fingerprint and bioactivity evaluation. The chemical fingerprints of 32 batches of S. miltiorrhizae samples from different sources were developed using high-performance liquid chromatography with diode array detection, and then were evaluated by similarity analysis and hierarchical clustering analysis. Anti-P. aeruginosa activity was determined by microcalorimetry. Some crucial thermokinetic parameters obtained from the heat-flow power-time curves of P. aeruginosa growth in the absence or presence of these S. miltiorrhizae samples were evaluated using principal component analysis. Thereafter, multiple linear regression analysis was used to analyze the fingerprint-activity relationship between the chemical fingerprints and anti-P. aeruginosa activity. This established the related equation between the inhibition ratio (I, %) of S. miltiorrhizae samples on P. aeruginosa and the peak areas of the common peaks. The results showed that the 32S. miltiorrhizae samples could be grouped into three clusters according to their chemical fingerprints and anti-P. aeruginosa activities. Protocatechualdehyde, salvianolic acid B, together with three unidentified compounds might be the major components that contributed largely to the antibacterial properties of S. miltiorrhizae and should be the focus of S. miltiorrhizae quality control. Thus, this study provided a preferred way for exploring the bioactive components of medicinal plants.
Highlights
Salvia miltiorrhizae, called “Danshen” in Chinese, has been officially listed in the Chinese pharmacopoeia (2015 edition), United States Pharmacopeia (USP39-NF34) and European Pharmacopoeia (EP6.1) as an important and well-known medicinal plant
The results showed that the relative standard deviation (RSD) values of peak areas for evaluating the precision, repeatability and stability of the method were less than 8.79%, 8.28% and 6.82% respectively
The results obtained from Principal component analysis (PCA) based on the anti-P. aeruginosa activities of S. miltiorrhizae samples were in accordance with that of hierarchical clustering analysis (HCA) of the chemical fingerprints
Summary
Salvia miltiorrhizae (family Lamiaceae), called “Danshen” in Chinese, has been officially listed in the Chinese pharmacopoeia (2015 edition), United States Pharmacopeia (USP39-NF34) and European Pharmacopoeia (EP6.1) as an important and well-known medicinal plant. This study combined the chemical fingerprints and anti-P. aeruginosa effects of S. miltiorrhizae to establish an integrated evaluation system for exploring the putative active components of this medicinal plant representing its biothermal activity. The chemical fingerprints and biothermal activities of S. miltiorrhizae from different sources were combined to establish fingerprint-activity relationships with the help of chemometrics by carrying out similarity analysis, principal component analysis and multiple linear regression analysis Understanding these relationships allowed us to further explore the active components of S. miltiorrhizae that contributed largely to the anti-P. aeruginosa activity. The approach described in this study provides a preferred model for future studies It illustrates how chemical fingerprints can be combined using LC method and biothermal activity by introducing microcalorimetry technique to study the active components of medicinal plants and other valuable materials
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