Discovery of the Potential Biomarkers for Discrimination between Hedyotis diffusa and Hedyotis corymbosa by UPLC-QTOF/MS Metabolome Analysis

Abstract

Hedyotis diffuse Willd. (HD) and Hedyotis corymbosa (L.) Lam. (HC), two closely related species of the same genus, are both used for health benefits and disease prevention in China. HC is also indiscriminately sold as HD in the wholesale chain and food markets. This confusion has led to a growing concern about their identification and quality evaluation. In order to further understand the molecular diversification between them, we focus on the screening of chemical components and the analysis of non-targeted metabolites. In this study, UPLC-QTOF-MSE, UNIFI platform and multivariate statistical analyses were used to profile them. Firstly, a total of 113 compounds, including 80 shared chemical constituents of the two plants, were identified from HC and HD by using the UNIFI platform. Secondly, the differences between two herbs were highlighted with the comparative analysis. As a result, a total of 33 robust biomarkers enabling the differentiation were discovered by using multivariate statistical analyses. For HC, there were 18 potential biomarkers (either the contents were much greater than in HD or being detected only in HC) including three iridoids, eight flavonoids, two tannins, two ketones, one alcohol and two monoterpenes. For HD, there were15 potential biomarkers (either the contents were much greater than in HC or being detected only in HD) including two iridoids, eight flavonoids, one tannin, one ketone, and three anthraquinones. With a comprehensive consideration of the contents or the MS responses of the chemical composition, Hedycoryside A and B, detected only in HC, could be used for rapid identification of HC. The compounds 1,3-dihydroxy-2-methylanthraquinone and 2-hydroxy-3-methylanthraquinone, detected only in HD, could be used for rapid identification of that plant. The systematic comparison of similarities and differences between two confusing Chinese herbs will provide reliable characterization profiles to clarify the pharmacological fundamental substances. HC should not be used as the substitute of HD.

Comprehensive metabolomics analysis based on UPLC-Q/TOF-MSEand the anti-COPD effect of different parts ofCelastrus orbiculatusThunb.

“…The optimized MS parameters were as follows: source temperature (150 C), desolvation temperature (400 C), cone voltage (40 V), capillary voltage at 2.6 kV (ESI + ) and 2.2 kV (ESI À ), cone gas ow (50 L h À1 ) and desolvation gas ow (800 L h À1 ). MS E mode was chosen with low energy of 6 V and high energy of 20-40 V. 26,27 The mass spectrometer was calibrated with sodium formate in the range of 100 to 1200 Da in order to ensure the mass reproducibility and accuracy. Leucine enkephalin (m/z 556.2771 in ESI + and 554.2615 in ESI À ) was used as external reference for Lock Spray™ injected at a constant ow of 10 mL min À1 .…”

UPLC-QTOF/MS-Based Nontargeted Metabolomic Analysis of Mountain- and Garden-Cultivated Ginseng of Different Ages in Northeast China

“…Main parameters were set as follows: peak intensity of high energy over 200 counts and the peak intensity of low energy over 1000 counts were the selected parameters in peak detection; mass error up to ±10 ppm for identified compound; retention time tolerance was set in the range of ±0.1 min; positive adducts containing +H, +Na or negative adducts containing −H, +HCOOH were all selected; the reference compound was leucine-enkephalin (556.2766 for positive ion, 554.2620 for negative ion). The MS raw data were processed using the streamlined workflow of UNIFI software to quickly identify the chemical components that met the match criteria with the in-house Traditional Medicine Library and the self-built database [20,21].…”

“…To differentiate MCG and CG, MarkerLynx XS V4.1 software (Waters, Milford, DE, USA) was used to process the raw data by deconvolution, alignment, data reduction and to perform the multivariate statistical analysis [20,21]. The following steps were performed: acquiring data, creating a MarkerLynx processing method, processing the acquired data and viewing results Extended Statistics (XS) Viewer.…”

“…Such analysis was also applied to make metabolite profiling and age discrimination of 4-and 6-year-old red ginseng [17], or 1-6 years ginseng [18].In addition, UNIFI, the automated data processing software, is an integrated informatics platform that possesses the ability to incorporate scientific library into a streamlined workflow, aiming at identifying chemical components from complex raw data [19]. The combination of UPLC separation, Q/TOF-MS detection and UNIFI platform has been frequently applied in the characterization of chemical constituents of herbs [20,21].Normally, CG is harvested after a 4-6 years cultivation period, and MCG is collected at ages of 10-20 years. To develop a more direct and more efficient discrimination method for the cultivation ages and to explore potential age-dependent markers, we chose 4, 5, 6-year-old CG and 12, 20-year-old MCG as the analytical samples in the present study.…”

“…In addition, UNIFI, the automated data processing software, is an integrated informatics platform that possesses the ability to incorporate scientific library into a streamlined workflow, aiming at identifying chemical components from complex raw data [19]. The combination of UPLC separation, Q/TOF-MS detection and UNIFI platform has been frequently applied in the characterization of chemical constituents of herbs [20,21].…”