Chemical characteristics of Rhodiola Crenulata and its mechanism in acute mountain sickness using UHPLC-Q-TOF-MS/MS combined with network pharmacology analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola crenulata (Hook.f. & Thomson) H.Ohba has a long history of clinical application for the prevention and treatment of acute mountain sickness (AMS) in traditional Chinese medicine. However, gaps in knowledge still exist in understanding the underlying mechanisms of Rhodiola crenulata against AMS. AIMS: To address this problem, a comprehensive method was established by combining UHPLC-Q-TOF-MS/MS analysis and network pharmacology. MATERIALS AND METHODS: The ingredients of Rhodiola crenulata were comprehensively analyzed using UHPLC-Q-TOF-MS/MS method. On this basis, a network pharmacology method incorporated target prediction, protein-protein interaction network, gene enrichment analysis and components-targets-pathways network was performed. Finally, the possible mechanisms were verified through molecular docking, in vitro and in vivo experiments. RESULTS: A total of 106 constituents of Rhodiola crenulata were charactered via UHPLC-Q-TOF-MS/MS. The 98 potentially active compounds out of 106 were screened and corresponded to 53 anti-AMS targets. Gene enrichment analysis revealed that hypoxia and inflammation related genes may be the central factors for Rhodiola crenulata to modulate AMS. Molecular docking revealed that TNF, VEGFA and HIF-1α had high affinities to Rhodiola crenulata compounds. Subsequently, Rhodiola crenulata extract was indicated to inhibit the protein expression level of TNF in hypoxia induced H9c2 cells. Lastly, Rhodiola crenulata extract was further verified to ameliorate heart injury and decreased the heart levels of TNF, VEGFA and HIF-1α in acute hypoxia-induced rats. CONCLUSIONS: This study used UHPLC-Q-TOF-MS/MS analysis and a network pharmacology to provide an important reference for revealing the potential mechanism of Rhodiola crenulata in the prevention and treatment of AMS.

The Study of Steaming Durations and Temperatures on the Chemical Characterization, Neuroprotective, and Antioxidant Activities of Panax notoginseng.

Panax notoginseng (PN) is one of the most valuable traditional Chinese medicines and has extensive pharmacological effects. Recent studies demonstrated that PN exhibited pharmacological effect related to Alzheimer's disease (AD). However, whether steaming process can boost its anti-AD activity is still unexplored. To fill this gap, effects of steaming durations and temperatures on the chemical characterization, neuroprotective and antioxidant activities of PN were systematically investigated in this study. HPLC fingerprint coupled with quantitative analysis demonstrated striking conversion of original saponins to less polar ones with the increase in the steaming time and temperature. In the viewpoint of anti-AD activity on neuroprotective and antioxidant effects, several steamed PN samples (110°C-6/8/10 h, 120°C -4/6 h samples) displayed a significant increase both in cell viability and oxygen radical absorption capacity (ORAC) values compared with the no steamed one (P < 0.01 or P < 0.005). Steaming temperature had the greater impact on the change of chemical composition and anti-AD activity of PN. Moreover, the spectrum-effect relationship analysis revealed that the transformed saponins were partially responsible for the increased neuroprotective and antioxidant effects of steamed PN. Therefore, steamed PN could be used as a potential crude drug for prevention and treatment of AD.

Application of UHPLC Fingerprints Combined with Chemical Pattern Recognition Analysis in the Differentiation of Six Rhodiola Species.

Rhodiola, especially Rhodiola crenulate and Rhodiola rosea, is an increasingly widely used traditional medicine or dietary supplement in Asian and western countries. Because of the phytochemical diversity and difference of therapeutic efficacy among Rhodiola species, it is crucial to accurately identify them. In this study, a simple and efficient method of the classification of Rhodiola crenulate, Rhodiola rosea, and their confusable species (Rhodiola serrata, Rhodiola yunnanensis, Rhodiola kirilowii and Rhodiola fastigiate) was established by UHPLC fingerprints combined with chemical pattern recognition analysis. The results showed that similarity analysis and principal component analysis (PCA) could not achieve accurate classification among the six Rhodiola species. Linear discriminant analysis (LDA) combined with stepwise feature selection exhibited effective discrimination. Seven characteristic peaks that are responsible for accurate classification were selected, and their distinguishing ability was successfully verified by partial least-squares discriminant analysis (PLS-DA) and orthogonal partial least-squares discriminant analysis (OPLS-DA), respectively. Finally, the components of these seven characteristic peaks were identified as 1-(2-Hydroxy-2-methylbutanoate) ß-D-glucopyranose, 4-O-glucosyl-p-coumaric acid, salidroside, epigallocatechin, 1,2,3,4,6-pentagalloyglucose, epigallocatechin gallate, and (+)-isolarisiresinol-4'-O-ß-D-glucopyranoside or (+)-isolarisiresinol-4-O-ß-D-glucopyranoside, respectively. The results obtained in our study provided useful information for authenticity identification and classification of Rhodiola species.

Quantitative fingerprinting based on the limited-ratio quantified fingerprint method for an overall quality consistency assessment and antioxidant activity determination of Lianqiao Baidu pills using HPLC with a diode array detector combined with chemometric methods.

Lianqiao Baidu pills are widely used herbal medicinal preparation that were analyzed to develop a quality consistency technique. The characteristic fingerprints of 28 batches of Lianqiao Baidu pill samples were established at five wavelengths and simultaneously assessed by using a limited-ratio quantified fingerprint method using 15 marker compounds. The principal component analysis and fingerprinting results were compared, and the qualitative classification of the samples by principal component analysis agreed with their quantitative evaluation by the limited-ratio quantified fingerprint method. Furthermore, the antioxidant activities of the samples were surveyed and determined using a 2,2-diphenyl-1-picrylhydrazyl radical-scavenging approach. A relationship between the common peaks in the fingerprints and the antioxidant activities was established using a partial least squares model. The relationship can be used both to determine the antioxidant activities of the Lianqiao Baidu pill preparations in vitro and as a reference for the selection of active constituents for sample quality classification. The classification results for the samples based on principal component analysis agreed with the quantitative evaluation by the limited-ratio quantified fingerprint method, which demonstrated that the method can be applied to the holistic quality control of traditional Chinese medicine and herbal preparations.

Comprehensively Evaluating the Quality Consistency of Liuwei Dihuang Pill by Five-Wavelength Overall Fused HPLC Fingerprints Combined with Simultaneous Determination and Antioxidant Activity.

A combination method of five-wavelength overall fused fingerprints and multi-component quantification by high-performance liquid chromatography coupled with diode array detection was developed to monitor the quality consistency of a Traditional Chinese Medicine (TCM), Liuwei Dihuang Pill (LDP). The validation results demonstrated that this method met the requirements of fingerprint analysis and quantification analysis with desired linearity, precision, accuracy, LOD and LOQ. In the fingerprint assessments, the five-wavelength overall fused fingerprints were estimated by the quantified ratio fingerprint method from qualitative and quantitative perspectives, achieved by an in-house developed software "Digitized Evaluation System for Super-Information Characteristics of TCM/HM Fingerprints 4.0". In order to holistically reflect the quality of LDP, simultaneous analysis of the content of five marker compounds was also carried out. The correlation results between quantified fingerprints and quantitative determination demonstrated that multi-component quantification could be replaced by quantified fingerprints. In addition, the fingerprint-efficacy was also established and confirmed through the partial least squares to visualize the quality information of LDP. The study demonstrated that five-wavelength overall fused fingerprints combined with antioxidant activity assay provided a reliable and efficient method for the assessment of quality of TCM and herbal preparations.

[Quality evaluation of Yinqiaojiedu tablets by three wavelength fusion fingerprint combined with six components quantitative determination and principal component analysis].

The fusion high performance liquid chromatographic fingerprint combined with quantification and principal component analysis (PCA) was successfully developed, which was applied to assess the quality consistency of 25 Yinqiaojiedu tablets (YQJDTs). The resolution index I was employed as an objective function to evaluate chromatographic conditions. The chromatographic fingerprints were established by reversed-phase high performance liquid chromatography. The wavelengths were set at 230, 279 and 327 nm, separately. Then, the three wavelength fusion fingerprint was developed by the technique of multi-wavelength fusion fingerprint. The linearity, precision, repeatability, stability and accuracy of the method were proved to meet the fingerprint analysis criteria. Subsequently, the systematic quantified fingerprint method was applied for integrative quality discrimination of YQJDTs from both qualitative and quantitative perspectives. The results showed that the quality of 25 YQJDTs from the two manufacturers were completely qualified and well differentiated. The contents of six components were determined simultaneously and combined with fingerprint analysis. It was used to evaluate the quality of YQJDT from the overall and partial points of view. In addition, the PCA of the fusion fingerprint data was applied to get the score plot. It can be clearly distinguished the 25 YQJDT samples from the two manufacturers. The method proposed in this study was comprehensive and reliable, which could be used for a valuable reference to scientifically evaluate and effectively control the quality of YQJDT.

Holistic evaluation of San-Huang Tablets using a combination of multi-wavelength quantitative fingerprinting and radical-scavenging assays.

The present study was designed to establish a multi-wavelength quantitative fingerprinting method for San-Huang Tablets (SHT), a widely used and commercially available herbal preparation, where high performance liquid chromatography (HPLC) with a diode array detector (DAD) was employed to obtain the fingerprint profiles. A simple linear quantitative fingerprint method (SLQFM) coupled with multi-ingredient simultaneous determination was developed to evaluate the quality consistency of the tested samples qualitatively and quantitatively. Additionally, the component-activity relationship between chromatographic fingerprints and total radical-scavenging capacity in vitro (as assessed using the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay) was investigated by partial least squares regression (PLSR) analysis to predict the antioxidant capacity of new samples from the chromatographic fingerprints and identify the main active constituents that can be used as the target markers for the quality control of SHT. In conclusion, the strategy developed in the present study was effective and reliable, which can be employed for holistic evaluation and accurate discrimination for the quality consistency of SHT preparations and other traditional Chinese medicine (TCM) and herbal preparations as well.

Capillary electrophoresis fingerprints combined with chemometric methods to evaluate the quality consistency and predict the antioxidant activity of Yinqiaojiedu tablet.

The quality consistency of Yinqiaojiedu tablets (YQJDTs) was monitored by extracting their electrophoretic fingerprint and marker compound data that were obtained using capillary electrophoresis. To select the suitable background electrolyte, wing-shape method was proposed. A background electrolyte composed of 103.1 mM boric acid, 51.6 mM sodium borate, 9.8 mM disodium hydrogen phosphate, and 15.6 mM sodium dihydrogen phosphate was used to separate compounds. Under the optimized conditions, the content of three marker compounds was determined in 25 YQJDT samples. The capillary electrophoresis fingerprints were developed simultaneously, then 25 samples from two manufacturers were clearly divided into two clusters based on the principal component analysis. In fingerprint assessments, systematic quantified fingerprint method was adopted for integrative quality discrimination of YQJDTs from both qualitative and quantitative perspectives, by which the qualities of 25 samples were well differentiated. In addition, partial least squares model was established to explore fingerprint-efficacy relationship between the fingerprints and the antioxidant activities in vitro, providing clinically useful information for quality control. The proposed method was reliable and comprehensive, which could be used for a valuable reference to monitor the quality consistency of traditional Chinese medicines.