Chemical composition analysis of Angelica sinensis (Oliv.) Diels and its four processed products by ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry combining with nontargeted metabolomics.

Angelica sinensis (Oliv.) Diels. has been used for women to enrich the blood, prevent and treat blood deficiency syndrome in Traditional Chinese Medicine for thousands of years. Wine-processed Angelica sinensis, soil-processed Angelica sinensis, oil-processed Angelica sinensis, and charred-processed Angelica sinensis are the most significant four processed products used in Chinese clinic. However, there have been few studies aimed at comparing their chemical differences. Ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry combining with nontargeted metabolomics was applied to investigate the diversity of processed products of Angelica sinensis. A total of 74 compounds with the variable importance in the projection value more than 1.5 and P less than 0.05 in ANOVA were highlighted as the compounds that contribute most to the discrimination of Angelica sinensis and four processed products. The results showed the metabolic changes between Angelica sinensis and its four processed products, there were 19 metabolites, 3 metabolites, 6 metabolites, and 45 metabolites were tentatively assigned in soil-processed Angelica sinensis, wine-processed Angelica sinensis, oil-processed Angelica sinensis, and charred-processed Angelica sinensis, respectively. These results suggested that the proposed metabolomics approach was useful for the quality evaluation and control of processed products of Angelica sinensis.

Chemical composition analysis of Schisandra chinensis fructus and its three processed products using UHPLC-Q-Orbitrap/MS-based metabolomics approach.

An ultra-high performance liquid chromatography coupled with quadrupole Orbitrap mass spectrometry (UHPLC-Q-Orbitrap/MS)-based metabolomics method was applied to investigate the chemome diversity of Schisandra chinensis fructus (SF) and its processed products, including vinegar-processed Schisandra (VS), wine-processed Schisandra (WS), and honey-processed Schisandra (HS). A clear classification among four Schisandra products was observed in the score plot of the partial least-squares discriminant analysis (PLS-DA) model, then 28 marker compounds were selected and identified. The content of most marker compounds in VS and WS was increased compared with that in SF, and the lowest content was observed in HS, then the high-performance liquid chromatography (HPLC) analysis was performed to confirm the change trends. These results suggested the chemical composition variation occurs in different Schisandra products, and the marker compounds selected in this study will be useful for the quality evaluation of Schisandra products.

Rapid characterization of Schisandra species by using direct analysis in real time mass spectrometry.

Direct analysis in real time ionization source coupled with quadrupole orbitrap mass spectrometry (DART-Q-Orbitrap MS) was applied to analyze the Schisandra chinensis (S. chinensis) and Schisandra sphenanthera (S. sphenanthera) samples. The experimental condition including the ionization gas and gas temperature were optimized to obtain the best performance. The DART-MS analysis was operated using helium at 250 °C. The partial least squares discriminant analysis (PLS-DA) was conducted based on the DART-MS data to explore the differences between S. chinensis and S. sphenanthera samples. The clear separation between groups was observed in the PLS-DA score plot, indicating the chemome diversity of these two samples. Then 8 compounds that contribute most to the sample classification were selected and annotated, and the intensity change tendency of these compounds was similar to that obtained by the high-performance liquid chromatography (HPLC) method. Besides, these two species can also be discriminated by examining the existence of the compound anwulignan at m/z 328.1656 in this study. Our results show that DART-MS is a powerful analytical tool with the merit of rapid analysis speed, easy to handle, low consumption of organic solvent, and has the great potential for rapid detection and discrimination of S. chinensis and S. sphenanthera. It is expected that the established method could provide a rapid, reliable method for the quality assessment of Schisandra species, and expand this method to the analysis of other herbal medicines.

Electrochemical detection of protein kinase activity based on carboxypeptidase Y digestion triggered signal amplification.

An effective assay method for monitoring protein kinase activity and screening inhibitors is greatly beneficial to kinase-related drug discovery, early diagnosis of diseases, and therapeutic effect evaluation. Herein, we develop a simple electrochemical method for detecting the activity of casein kinase II (CK2) based on phosphorylation against carboxypeptidase Y (CPY) digestion triggered signal amplification, where CK2 catalyzed phosphorylation event protects the substrate peptide from the digestion of CPY, maintains the repulsive force of the substrate peptide towards the redox probe, and results in a weak electrochemical signal. Whereas, without phosphorylation, the substrate peptide is digested by CPY and a strong electrochemical signal is obtained. The detection feasibility is demonstrated for the assay of CK2 activity with low detection limit of 0.047unit/mL. Moreover, the biosensor was used for the analysis of kinase inhibition. Based on the electrochemical signal dependent inhibitor concentration, the IC50 value of ellagic acid was estimated to be 39.77nM. The proposed method is also successfully applied to analyze CK2 activity in cell lysates, proving the applicability in complex biological samples.

Rapid identification of traditional Chinese herbal medicine by direct analysis in real time (DART) mass spectrometry.

Direct analysis in real time-mass spectrometry (DART-MS) was employed as a novel fast method to identify traditional Chinese herbal medicine (TCHM). In order to obtain high quality mass spectra, the ionization temperature was optimized for every kind of sample. With minimal or no sample pretreatment, major TCHM components, including alkaloids, flavonoids and some ginsenosides, were directly detected within several seconds, while thirteen ginsenosides need derivatization to get good mass spectra. Pseudoginsenoside F11, compound K, protopanaxatriol (PPT) and protopanaxadiol (PPD), for the first time were detected without derivatization. Among five of eight tested Chinese herbal medicines, Rhizoma Corydalis, Bulbus Fritillariae Thunbergii, Arecae Semen, Ramulus Uncariae Cum Uncis and Scutellariae Radix, were first time identified by DART-MS. In addition, the ionization mechanisms of major herbal components, alkaloids, flavonoids and ginsenosides, were discussed in detail. Our results demonstrated that DART-MS could provide a rapid, reliable and environmental friendly method for the rapid identification of TCHM, and may be applicable to other plants.