Search

Abstract Natural products-coordinated metal ions to form the nanomedicines are in the spotlight for cancer therapy. Some natural products could be coordinated with metal ions forming nanomedicines via simple and green environmental self-assembly, which not only improved the bioavailability of natural products, but also conferred multiple therapeutic modalities and multimodal imaging. On the one hand, in the weak acidity, glutathione (GSH) and hydrogen peroxide (H2O2) overexpression of tumor microenvironment (TME), such carrier-free nanomedicines could be further enhanced the therapeutic effect via optimizing the species of metal ions. On the other hand, nanomedicines could exert the precise treatment of tumor under the guidance of multiple imaging. Hence, this review summarized the research progress in recent years on the application of natural product-coordinated metal ions in cancer therapy. In addition, the prospects and challenges for the application of natural product-coordinated metal ions were discussed, especially how to improve targeting ability and stability and assess the safety of metal ions, so as to facilitate the clinical translation and application of natural product-coordinated metal ions nanomedicines. Graphical Abstract

Background: Viticis Fructus (also known as Manjingzi) has been used in China for more than 2000 years. It is one of the most famous traditional Chinese medicines with the main effect of dispelling wind-heat. In Asian countries, it is used in the treatment of common cold with wind-heat syndrome, swollen gums, migraines, dizziness, etc. The aim of the review: The paper emphasizes on the botany, historical records, phytochemistry, pharmacology, toxicity, quality control, pharmacokinetics and comprehensive applications of Viticis Fructus to furnish with a scientific theoretical reference for its exploration and applications. Materials and methods: Correlative data on Viticis Fructus were obtained from PubMed, ScienceDirect, Web of science, Embase, Scopus, Google Scholar, CNKI, WeiPu, Chinese ancient books and DuXiu academic search. The data collection ended in May 2024. Results: The results showed 324 compounds, including terpenoids, flavonoids, lignans and others, which were isolated and identified from Viticis Fructus. In addition to treating headaches and eye pain, Viticis Fructus also had anti-inflammatory, antiplatelet activation, analgesia, antitumor and antihypertensive effects. In addition, this review summarized the botany, toxicity, counterfeit identification, pharmacokinetics and patent information of Viticis Fructus in detail. The shortcomings and feasible suggestions are put forward, which provide a basis for further research and utilization of Viticis Fructus. Conclusion: Viticis Fructus has been used to remedy common cold with wind-heat syndrome, cephalalgia, ophthalmodynia, dizziness, etc. The pharmacological activities of the main components have been clarified and backed up the traditional applications. However, the botany, processing, pharmacological mechanism, quality control and toxicological studies of Viticis Fructus need to be further improved.

Abstract Cimicifugae Rhizoma, generally known as “Sheng Ma” in China, has great medicinal and dietary values. Cimicifugae Rhizoma is the dried rhizome of Cimicifuga foetida L., Cimicifuga dahurica (Turcz.) Maxim. and Cimicifuga heracleifolia Kom., which has been used to treat wind-heat headache, tooth pain, aphtha, sore throat, prolapse of anus and uterine prolapse in traditional Chinese medicine. This review systematically presents the traditional uses, phytochemistry, pharmacology, clinical studies, quality control and toxicity of Cimicifugae Rhizoma in order to propose scientific evidence for its rational utilization and product development. Herein, 348 compounds isolated or identified from the herb are summarized in this review, mainly including triterpenoid saponins, phenylpropanoids, chromones, alkaloids, terpenoids and flavonoids. The crude extracts and its constituents had various pharmacological properties such as anti-inflammatory, antitumor, antiviral, antioxidant, neuroprotective, anti-osteoporosis and relieving menopausal symptoms. The recent research progress of Cimicifugae Rhizoma in ethnopharmacology, phytochemistry and pharmacological effects demonstrates the effectiveness of its utilization and supplies valuable guidance for further research. This review will provide a basis for the future development and utilization of Cimicifugae Rhizoma.

Background: Scutellariae radix (SR) is the dried root of Scutellaria baicalensis Georgi. It has a long history of ethnic medicinal use, traditionally recognized for its efficacy in clearing heat， drying dampness, eliminating fire， removing toxins, stopping bleeding and tranquilizing fetus to prevent miscarriage. Clinically, it is used to treat cold, fever, migraine, hand-foot-and-mouth diseases, liver cancer and inflammatory diseases. Purpose: The review aims to provide a comprehensive reference on the ethnobotanical uses, processing, phytochemistry, pharmacological effect, quality control and influence factors of biosynthesis for a deeper understanding of SR. Results and conclusion: A total of 210 isolated components have been reported in the literature, including flavonoids and their glycosides, phenylpropanoids, phenylethanoid glycosides, phenolic acids, volatile components, polysaccharides and others. The extract of SR and its main flavonoids such as baicalin, baicalein, wogonin, wogonoside, and scutellarin showed antioxidant, anti-inflammatory, anti-tumor, antiviral, hepatoprotective, and neuroprotective effects. However, further studies are required to elucidate its mechanisms of action and clinical applications. The pharmacodynamic evaluation based on traditional efficacy should be conducted. Although various analytical methods have been established for the quality control of SR, there are gaps in the research regarding efficacy-related quality markers and the development of quality control standards for its processed products. The regulatory mechanisms of flavonoids biosynthesis remain to be explored while the influence of environmental and transcription factors on the biosynthesis have been studied. In conclusion, SR is a promising herbal medicine with significant potential for future development.

Pogostemon cablin (PC) is a traditional Chinese medicine (TCM) and food as well as an important essential oil plant in China. PC essential oil exerts pharmacological effects such as anti-inflammatory, anti-oxidant, anti-platelet, anti-thrombotic, and anti-depressant. This study established a reliable and sensitive gas chromatography-mass spectrometry (GC-MS) method for the simultaneous determination of the pharmacokinetics of patchouli alcohol, β-elemene, β-caryophyllene, caryophyllene oxide, and farnesol in the plasma of rats after oral administration of PC essential oil extract. Using ethyl acetate to prepare the plasma samples, and p-menthone was used as the internal standard (IS). An HP5-MS column (0.25 μm × 0.25 mm × 30 m) was used for chromatographic separation, and detection was performed in selected ion monitoring (SIM) mode. The accuracies of intra-day and inter-day for all analytes displayed a range of −6.7 %–9.2 %, with precision below 12.5 %. Extraction recoveries for analytes ranged from 74.0 to 106.4 % and matrix effects ranged from 92.4 to 106.9 %. Stability results have demonstrated that the relative standard deviations (RSD) of analytes were below 12.1 %. Therefore, the developed GC-MS method successfully evaluated the pharmacokinetics of five volatile components in PC essential oil extract administered orally to rats.

Introduction:Pogostemon cablin (PC) is used in traditional Chinese medicine and food, as it exerts pharmacological effects, such as immune-modulatory, antibacterial, antioxidant, antitumor, and antiviral. Currently, the pharmacokinetics (PK) studies of PC mainly focus on individual components. However, research on these individual components cannot reflect the actual PK characteristics of PC after administration. Therefore, the simultaneous determination of multiple components in rat plasma using UPLC-MS/MS was used for the pharmacokinetic study after oral administration of PC extract in this study, providing reference value for the clinical application of PC.Methods: In the present study, a reliable and sensitive ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of 15 prototype components (vanillic acid, vitexin, verbascoside, isoacteoside, hyperoside, cosmosiin, apigenin, β-rhamnocitrin, acacetin, ombuin, pogostone, pachypodol, vicenin-2, retusin, and diosmetin-7-O-β-D-glucopyranoside) in rat plasma after oral administration of the PC extract. Plasma samples were prepared via protein precipitation using acetonitrile, and icariin was used as the internal standard (IS).Results: The intra-day and inter-day accuracies ranged from −12.0 to 14.3%, and the precision of the analytes was less than 11.3%. The extraction recovery rate of the analytes ranged from 70.6−104.5%, and the matrix effects ranged from 67.4−104.8%. Stability studies proved that the analytes were stable under the tested conditions, with a relative standard deviation lower than 14.1%.Conclusion: The developed method can be applied to evaluate the PK of 15 prototype components in PC extracts of rats after oral administration using UPLC-MS/MS, providing valuable information for the development and clinical safe, effective, and rational use of PC.

The multiple species characteristics of traditional Chinese medicines (TCMs) are crucial for expanding TCMs sources, meeting the needs of the pharmaceutical industry and ensuring clinical requirements. It’s also one of the significant factors affecting the quality control of TCMs. Systematic differential analysis of original species in TCMs is an important link in achieving comprehensive quality control, ensuring the effectiveness and safety of clinical medication. The study aims to establish a reliable and efficient approach to screen combinatorial discriminatory quality markers for rapid differentiation of original species by metabolomics coupled with DNA barcoding as a case of Cimicifugae Rhizoma. DNA barcoding is used to identify the origin of Cimicifugae Rhizoma. The data-dependent acquisition mode integrated with the computerized intelligent filtering system was established for in-depth characterization of metabolites from Cimicifugae Rhizoma using ultra-high performance liquid chromatography to quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). The untargeted metabolomics combined with multivariate statistical analysis was performed to screen and identify the potential combinatorial discriminatory quality markers. Finally, quantitative analysis and predictive model of these markers were employed to validate the feasibility of this strategy to distinguish the original species. Based on the scores of variable importance in projection greater than 1.0 and t-test (p

Many different species of Rhodiola are traditional botanical medicines with various effects, and their roots and rhizomes are the medicinal parts, which have been recorded in many ancient medical books. The 2020th edition of Chinese Pharmacopoeia stipulates Rhodiola crenulata (Hook. f. et Thoms.) H. Ohba (R. crenulata) as the only medicinal species. However, the wild resources of R. crenulata are gradually decreasing, and it is difficult to achieve artificial cultivation of R. crenulata nowadays. It is important to find several alternative species of R. crenulata. In the present study, an integrated strategy based on chemical profiling, disease module-guided network pharmacology, and quantitative analysis (CDMQ) has been developed to estimate the chemical characteristics, determine active components and related disease modules, discover potential quality markers, and identify alternative species in a step-by-step manner for 9 species of Rhodiola. Firstly, a total of 109 compounds were identified in accordance with their retention times, accurate masses and characteristic MS/MS fragment patterns, which mainly included phenylpropanoid glycosides, gallic acid glycosides, flavonoids and their glycosides, flavanols, organic acids, and others. Subsequently, disease module-guided network pharmacology analysis was performed based on the related targets of 109 compounds. In combination of the limit of detection of the UPLC-QQQ-MS/MS, 12 compounds were identified as active components. The enriched disease modules of circulatory system, mouth, respiratory system, skin and connective tissue showed close relationship with traditional efficacy of Rhodiola species. It also revealed that salidroside, rhodiosin, epicatechin gallate, catechin gallate, and 6′-O-galloylsalidroside changed significantly in different species to be served as potential quality markers for Rhodiola. Among the 9 species, Rhodiola kirilowii (Regel) Maxim (R. kirilowii) and Rhodiola sacra (Prain ex Hamet) S.H. Fu (R. sacra) could probably be used as an alternative species of R. crenulata. Overall, this study established a solid foundation and provided theoretical guidance for the chemical profiling, prediction of clinical application, and quality control of different Rhodiola species.

Valeriana jatamansi Jones (V. jatamansi) is an ethnomedicinal herb widely used worldwide for its excellent anti-inflammatory activity. However, the quality markers that were correlated with the anti-inflammatory activity in V. jatamansi remained unclear. This study aimed to identify combinatorial quality markers based on the anti-inflammatory effects of V. jatamansi for enhancing its quality control. The chemical constituents of V. jatamansi were identify by UPLC-Q-TOF-MS, and 36 compounds were identified. The anti-inflammatory mechanism was predicted by network pharmacology, STAT3 and JAK1 were selected as key targets. The anti-inflammatory activity and predicted targets were validated by using lipopolysaccharide (LPS)-stimulated RAW264.7 cells in vitro. A combination of bivariate correlation analysis (BCA) and grey correlation analysis (GCA) was applied to evaluate the correlation between the chemical spectrum and the anti-inflammatory activities. Hesperidin, acevaltrate and valtrate were screened as potential quality markers. These three compounds exhibited anti-inflammatory effects by inhibiting the production of inflammatory mediators (NO, IL-6, IL-1β and TNF-α) and suppressing the expression of the two key targets (JAK1 and STAT3). The quantitative analysis results showed that the contents of the combinatorial quality markers were positively correlated with the anti-inflammatory activity of V. jatamansi. The contribution rate of the combinatorial quality markers reached 45.0 %. This study provides a comprehensive strategy for the quality control of V. jatamansi based on combinatorial quality markers.

Weichang'an pill (WCAP) is a traditional Chinese patent medicine, which is clinically used for the treatment of bowel syndrome and functional dyspepsia such as diarrhea, abdominal distension, and enteritis. So far, quality control studies of WCAP have mainly focused on the determination of chemical composition content, which has little relevance to biological activity and clinical effects. With the aim of identifying the multi-index ingredients with NF-κB inhibitory activities related to WCAP clinical effect, this present work described the chemical profile of WCAP by UHPLC-QE-MS, established the correlated relationship between chromatographic fingerprints and the NF-κB inhibitory activities based on multivariate statistical analysis, including hierarchical clustering analysis (HCA), Pearson correlation analysis, and Partial least squares regression analysis (PLSR). The spectrum-effect relationship analysis indicated 10 compounds, which were ferulic acid, naringin, narirutin, hesperidin, neohesperidin, aloe emodin, emodin, honokiol, magnolol, and physcion, might be the potential NF-κB inhibitory constituents in the pill. The NF-κB inhibitory effects of the ten compounds were verified by in vitro dual luciferase reporting detection system. Considering that the detection index should be representative of more medicinal materials, a rapid and efficient UPLC-DAD method was eventually developed to determine the content of the 13 components. Our findings will provide data support for WCAP quality control and advance the understanding of the quality assessment of traditional Chinese patent medicines.

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as “Guang Dilong” and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 “decoction pieces” samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

In terrestrial medicinal plants, polyacetylenoids have been isolated from almost 110 species belonging to 11 families including Compositae, Apiaceae, Araliaceae, Campanulaceae, Annonaceae, and Meliaceae. They are a class of natural products derived from fatty acids with carbon chain lengths of C8–19, C21, C23–25, C27, C29, and C33, possessing a pleiotropic profile of bioactivities such as anti-tumor, anti-inflammatory activities. Herein, this review aims at summarizing the inventory of polyacetylenoids occurring in terrestrial medicinal plants during the last two decades from 2000 to 2023, the NMR characteristics, and the progress on analytical methods and pharmacological investigation of the well-known plant polyacetylenoids.

As a Traditional Chinese Medicine prescription, Qingjin Yiqi Granules (QJYQ) provides an effective treatment for patients recovering from COVID-19. However, the pharmacokinetics characteristics of the main components of QJYQ in vivo are still unknown. An efficacious ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated for the simultaneous determination of 33 components in rat plasma after oral administration of QJYQ. The plasma samples were precipitated with 400 µL methanol/acetonitrile (1/1, v/v) and analyzed in scheduled multiple reaction monitoring mode. The linear relationship of the 33 components was good (r > 0.9928). The lower limit of quantification for 33 components ranged from 0.4–60.5 ng/mL. The average recoveries and matrix effects of the analytes ranged from 72.9% to 115.0% with RSD of 1.4%–15.0%. All inter-day and intra-day RSDs were within 15.0%. After oral administration (3.15 g/kg), the validated approach was effectively applied to the pharmacokinetics of main components of QJYQ. Finally, fifteen main constituents of QJYQ with large plasma exposure were obtained, including baicalin, wogonoside, wogonin, apigenin-7-O-glucuronide, verbenalin, isoferulic acid, hesperidin, liquiritin, harpagide, protocatechuic acid, p-Coumaric acid, ferulic acid, sinapic acid, liquiritin apioside and glycyrrhizic acid. The present research lays a foundation for clarifying the therapeutic material basis of QJYQ and provides a reference for further scientific research and clinical application of QJYQ.

Veratrum nigrum L. (VN) is a well-known herbal medicine and rich in chemical components with multiple pharmacological activities including antihypertensive, anticancer, and antifungal effects. In the current experiment, the quality of VN from different habitats was evaluated based on combinative method of fingerprint, multi-component quantification and chemical pattern recognition. Fifteen batches of VN were collected, and intrinsic chemical composition were identified using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, which is a method for analyzing the similarity between samples, coupled with fingerprint of traditional Chinese medicine. The fingerprint similarity model show that 22 common peaks were selected covering 15 batches of and the similarity > 0.963. The total of 22 joint components were tentatively identified by comparison with standard substances or literature. A ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry method for simultaneous determination of 8 compounds was established to evaluate the contents of raw and processed Veratrum nigrum L. Multivariate analysis was then applied to compare different batches of herbs based on ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry data. All raw and processed samples were classified by partial least squares discriminant analysis based on the 8 analyzed compounds. The findings suggested that veratramine and polydatin with a variable importance for the project (VIP) > 1 were identified as significant constituents, the presence of which can be used to differentiate between raw and processed Veratrum nigrum L. samples. These results indicate that processing methods show important effects on the composition of Veratrum nigrum L..

A simple and efficient vortex-assisted matrix solid phase dispersion with a ultra-high-performance liquid chromatography-triple quadrupole mass spectrometer (VA-MSPD-UHPLC-MS/MS) was applied for simultaneous extraction and determination of seven alkaloids and three organic acids from Uncariae Ramulas Cum Unicis. The optimal extraction conditions of the target components were obtained by Box-Behnken design (BBD) combined with response surface methodology (RSM). The results of the method validation showed that this analytical method displayed good linearity with a correlation coefficient (r) no lower than 0.9990. The recoveries of ten active ingredients from Uncariae Ramulas Cum Unicis ranged from 95.9% to 103% (RSD ≤ 2.77%). The RSDs of intra-day and inter-day precisions were all below 2.97%. The present method exhibited not only lower solvent and sample usage, but also shorter sample processing and analysis time. Consequently, the developed VA-MSPD-UHPLC-MS/MS method could be successfully and effectively used for the extraction and analysis of ten active components from Uncariae Ramulas Cum Unicis.

Terminalia chebula Retz. (TC) is a well-known Chinese herbal medicine and rich in chemical components with multiple pharmacological effects. In this study, an ultra-performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) method was developed and used to determine the blood concentrations of nine active compounds (chebulic acid, gallic acid, protocatechuic acid, corilagin, chebulagic acid, chebulinic acid, 1,2,3,4,6-O-pentagalloylglucose, ellagic acid and ethyl gallate) after oral administration of TC extracts in rats. Pretreatment of plasma samples with protein precipitate with methanol was carried out, and caffeic acid was used as the internal standard (IS). Compounds precisions of intra- and inter-day were less than 14.6%, and the accuracy ranged from −11.7% to 13.5%. The extraction recoveries of compounds were between 84.9% and 108.4%, while matrix effects occurred between 86.4% and 115.9%. Stability tests showed that all nine analytes had been stable under four storage conditions, and statistically significant the relative standard deviations were under 13.7%. The validated UPLC-MS/MS method was applied with great success to plasma pharmacokinetics analysis of the TC extracts, and the pharmacokinetic results showed that among the nine components, the area under the concentration-time curve (AUC(0-tn), 231112.38 ± 64555.20 h ng/mL) and maximum concentration (Cmax, 4,983.57 ± 1721.53 ng/mL) of chebulagic acid were relatively large, which indicated that it had a higher level of plasma exposure. The half-life of elimination (T1/2) of chebulinic acid, corilagin and chebulagic acid were 43.30, 26.39 and 19.98 h, respectively, suggesting that these analytes showed prolonged retention and metabolize more slowly in vivo. This study would deliver a theoretical foundation for the further application of TC in clinical practice.

Prunus mume fructus (MF) is used in traditional Chinese medicine and food, as it exerts pharmacological effects, such as antibacterial, antioxidant, antitumour, thirst-relieving, and antidiarrheal effects. In the present study, a reliable and sensitive ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of 16 prototype components (L-(-)-malic acid, 3,4-dihydroxybenzaldehyde, protocatechuic acid, vanillic acid, caffeic acid, D-(-)-quinic acid, citric acid, ferulic acid, syringic acid, cryptochlorogenic acid, neochlorogenic acid, chlorogenic acid, amygdalin, maslinic acid, corosolic acid, and rutin) in rat plasma after oral administration of the MF extract. Plasma samples were prepared via protein precipitation using acetonitrile. The 16 components were separated on an ACQUITY UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm) with a gradient mobile phase system of methanol and 0.1% (v/v) formic acid aqueous solution at a flow rate of 0.3 ml/min. All components were quantitated using Agilent Jet Stream electrospray ionisation in negative ion mode. The intra-day and inter-day accuracies ranged from-9.4 to 9.4%, and the precision of the analytes was less than 14.8%. The extraction recovery rate of the analytes ranged from 63.59 to 109.44% and the matrix effects ranged from 49.25 to 109.28%. Stability studies proved that the analytes were stable under the tested conditions, with a relative standard deviation lower than 13.7%. Hence, the developed method was successfully applied to evaluate the pharmacokinetics of 16 components in the MF extract after oral administration in rats using UPLC-MS/MS.

The root of Panax notoginseng, a highly valued medicine and functional food, is the main part used for medicinal purposes. However, the stems and leaves are also used in practice. To provide a chemical basis for various uses, a quantitative comparison of 18 saponins using a non-targeted metabolomics approach was established, so as to investigate the chemical profiles of the different parts of P. notoginseng. The established strategy revealed that roots and stems, with their similar chemical characteristics, consisted mainly of protopanaxatriol-type saponins, whereas protopanaxadiol-type saponins were principally present in the leaves. Multivariate analysis further suggested that the quality of the stems and leaves of P. notoginseng was significantly affected by its geographical origin. Furthermore, 52 constituents (26 non-volatile and 26 volatile) were identified as potential markers for discriminating between different parts of the plant. Taken together, the study provides comprehensive chemical evidence for the rational application and exploitation of different parts of P. notoginseng.

Pheretima, one of the animal-derived traditional Chinese medicines, has been wildly used in various cardiovascular and cerebrovascular diseases, including stroke, coronary heart disease, hyperlipidemia, and hyperglycemia. However, it was still a big challenge to select the quality markers for Pheretima quality control. The fingerprint and network pharmacology-based strategy was proposed to screen the efficiency related quality markers (Q-Markers) of Pheretima. The ratio of sample to liquid, ultrasonic-extraction time, temperature, and power were optimized by orthogonal design, respectively. The chemical fingerprint of forty batches of Pheretima was established, and six common peaks were screened. The network pharmacology was used to construct the Pheretima-Components-Targets-Pathways-Stroke network. It was found that six potential efficacy Q-markers in Pheretima could exert the relaxing meridians effect to treat stroke through acting on multiple targets and regulating various pathways. A simple HPLC-DAD method was developed and validated to determine the efficacy Q-markers. Grey relational analysis was used to further verify the relation of potential efficiency related quality markers with the anticoagulation activity of Pheretima, which indicated that the contents of these markers exhibited high relationship with the anticoagulation activity. It was concluded that hypoxanthine, uridine, phenylalanine, inosine, guanosine, and tryptophan were selected as quality markers related to relaxing meridians to evaluate the quality of Pheretima. The fingerprint and network pharmacology-based strategy was proved to be a powerful strategy for the discovery of efficiency related Q-markers of Pheretima.

Abstract Cardiac hypertrophy is a pivotal pathophysiological step of various cardiovascular diseases, which eventually leads to heart failure and death. Extracts of Rhodiola species (Ext.R), a class of commonly used medicinal herbs in Europe and East Asia, can attenuate cardiac hypertrophy both in vitro and in vivo. Serum/glucocorticoid regulated kinase 1 (SGK1) is identified as a potential target of Ext. R. By mass spectrometry‐based kinase inhibitory assay, herbacetin (HBT) from Ext.R is identified as a novel SGK1 inhibitor with IC50 of 752 nmol. Thermal shift assay, KINOMEscan in vitro assay combined with molecular docking proves a direct binding between HBT and SGK1. Site‐specific mutation of Asp177 in SGK1 completely ablates the inhibitory activity of HBT. The presence of OH groups at the C‐3, C‐8, C‐4’ positions of flavonoids is suggested to be favorable for the inhibition of SGK1 activity. Finally, HBT significantly suppresses cardiomyocyte hypertrophy in vitro and in vivo, reduces reactive oxygen species (ROS) synthesis and calcium accumulation. HBT decreases phosphorylation of SGK1 and regulates its downstream forkhead box protein O1 (FoxO1) signaling pathway. Taken together, the findings suggest that a panel of flavonoids structurally related to HBT may be novel leads for developing new therapeutics against cardiac hypertrophy.

Geo-authentic herbs refer to medicinal materials produced in a specific region with superior quality. Stephania tetrandra S. Moore (S. tetrandra) is cultivated in many provinces of China, including Anhui, Zhejiang, Fujian, Jiangxi, Hunan, Guangxi, Guangdong, Hainan, and Taiwan, among which Jiangxi is the geo-authentic origin. To explore habitat-related chemical markers of herbal medicine, an integrated chromatographic technique including gas chromatography-mass spectrometry (GC-MS), ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) and ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) combined with chemometric analysis was established. The established methods manifested that they were clearly divided into two groups according to non-authentic origins and geo-authentic origins, suggesting that the metabolites were closely related to their producing areas. A total of 70 volatile compounds and 50 non-volatile compounds were identified in S. tetrandra. Meanwhile, tetrandrine, fangchinoline, isocorydine, magnocurarine, magnoflorine, boldine, and higenamine as chemical markers were accurately quantified and suggested importance in grouping non-authentic origins and geo-authentic origins samples. The discriminatory analysis also indicated well prediction performance with an accuracy of 80%. The results showed that the multiple chromatographic and chemometric analysis technique could be used as an effective approach for discovering the chemical markers of herbal medicine to fulfill the evaluation of overall chemical consistency among samples from different producing areas.

Sanye Tablet (SYT) is a patent prescription widely used in treating T2D and pre-diabetes, especially T2D comorbid with hypertriglyceridemia, for many years in China. However, the underlying mechanism that accounts for the anti-diabetic potential of SYT by regulating lipid-related intermediates remains to be elucidated. This study aimed to investigate the mechanism of SYT on lipid metabolism and insulin sensitivity in high-fat diet (HFD)-induced obese mice by means of combining lipidomics and proteomics. The obese mice models were developed via HFD feeding for 20 consecutive weeks. Mice in the treatment group were given metformin and SYT respectively, and the effects of SYT on body weight, blood glucose, insulin sensitivity, fat accumulation in the organs, and pathological changes in the liver were monitored. Lipid metabolism was examined by lipidomics. Further determination of signaling pathways was detected by proteomics. The biological contributions of the compounds detected in SYT’s chemical fingerprint were predicted by network pharmacology. SYT treatment reduced body weight, inhibited viscera and hepatic steatosis lipid accumulation, and prevented insulin resistance. Furthermore, it was found that circulatory inflammatory cytokines were reduced by SYT treatment. In addition, lipidomics analysis indicated that SYT targets lipid intermediates, including diacylglycerol (DAG) and Ceramide (Cer). Mechanistically, SYT positively affected these lipid intermediates by suppressing liver lipogenesis via downregulation of SREBP1/ACC and the JAK/STAT signaling pathway. Our results predicted that astragalin and rosmarinic acid might regulate the JAK-STAT pathway by targeting PIM2 and STAT1, respectively, while paeoniflorin and rosmarinic acid were likely to regulate inflammatory responses by targeting TNFα, IL-6, and IL-4 during T2D. Overall, our study provides supportive evidence for the mechanism of SYT’s therapeutic effect on dysregulated lipid metabolism in diabesity.

Mume Fructus is a well-known herbal medicine and food with a long history of processing and application. Different processing methods impact the intrinsic quality of Mume Fructus. Thus, it is of great significance to investigate the changes in chemical components during processing (i.e., raw compared to the pulp and charcoal forms). In this study, plant metabolomics methods based on mass spectrometry detection were established to analyze the chemical ingredients of Mume Fructus comprehensively. Chemometric strategies were combined to analyze the profile differences of Mume Fructus after different processing methods. The established strategy identified 98 volatile and 89 non-volatile compounds of Mume Fructus by gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS/MS), respectively. Moreover, the orthogonal partial least squares discriminant analysis (OPLS-DA) indicated that raw Mume Fructus and the Mume Fructus pulp and charcoal were distributed in three regions. Subsequently, 19 volatile and 16 non-volatile components were selected as potential chemical component markers with variable importance in the projection using (VIP) >1 as the criterion, and the accuracy was verified by a Back Propagation Neural Network (BP-NN). To further understand the difference in the content of Mume Fructus before and after processing, 16 non-volatile chemical component markers were quantitatively determined by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS). The results revealed that, compared with raw Mume Fructus, the total content of 16 components in the pulp of Mume Fructus increased while it decreased in the charcoal. Therefore, this study used GC-MS, UHPLC-Q-TOF-MS/MS and UHPLC-MS/MS modern technology to analyze the differences in chemical components before and after the processing of Mume Fructus and provided a material basis for further research on the quality evaluation and efficacy of Mume Fructus.

Asarum (Aristolochiaceae) is one of the common herbs used to relieve exterior syndromes. Some volatile components of Asarum which have toxic effect may cause adverse reactions such as headache, general tension, unconsciousness, and respiratory paralysis. Therefore, Asarum is normally processed to reduce such toxicity and adverse effects. The bioactive ingredients contained in different Asarum herbs vary significantly; this variation may be attributed to their differences in species, origins, or processing methods. In this study, 16 batches of Asarum herbs were collected, and their species were identified using DNA barcoding, which is a method for distinguishing plant species, coupled with microscopy. A gas chromatography-mass spectrometry (GC-MS) method for simultaneous determination of 10 compounds was established to evaluate the contents of raw and processed Asarum herbs. Multivariate analysis was then applied to compare different batches of herbs based on the GC-MS data. DNA barcoding identified the herbs as being derived from four sources, and herbs from different origins showed different microscopic features. The results demonstrated that most of the samples were clearly clustered into distinct groups that corresponded to species types. All raw and processed samples were classified by partial least squares discriminant analysis (PLS-DA) based on the 10 analyzed compounds. The findings suggested that safrole and methyleugenol with a variable importance in the project (VIP) > 1 are unique compounds that can be used to differentiate between Asarum species. Safrole, methyleugenol, and 2,6,6-trimethylcyclohepta-2,4-dien-1-one were identified as significant constituents, the presence of which can be used to differentiate between raw and processed Asarum samples. These results indicate that species and processing methods show important effects on the composition of Asarum herbs.

Quality assessment of Cortex Periplocae remains a challenge, due to its complex chemical profile. This study aims to investigate the chemical components of Cortex Periplocae, including its non-volatile and volatile constituents, via liquid chromatograph−mass spectrometry (LC-MS/MS) and gas chromatography−mass spectrometry (GC-MS) assays. The established strategy manifested that Cortex Periplocae from different producing areas was determined by identifying 27 chemical markers with ultra-high-performance liquid chromatography, coupled with quadrupole tandem time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS), including four main groups of cardiac glycosides, organic acids, aldehydes, and oligosaccharides. These groups’ variable importance in the projection (VIP) were greater than 1. Simultaneously, the samples were divided into four categories, combined with multivariate statistical analysis. In addition, in order to further understand the difference in the content of samples from different producing areas, nine chemical markers of Cortex Periplocae from 14 different producing areas were determined by high performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS), and results indicated that the main effective constituents of Cortex Periplocae varied with places of origin. Furthermore, in GC-MS analysis, samples were divided into three groups with multivariate statistical analysis; in addition, 22 differential components whose VIP were greater than 1 were identified, which were principally volatile oils and fatty acids. Finally, the relative contents of seven main volatile constituents were obtained, which varied extremely with the producing areas. The results showed that the LC-MS/MS and GC-MS assays, combined with multivariate statistical analysis for Cortex Periplocae, provided a comprehensive and effective means for its quality evaluation.

An efficient microwave-assisted extraction (MAE) combined with in-capillary [Fe(ferrozine)3]2+-capillary electrophoresis-Diode Array Detector (in-capillary [Fe(ferrozine)3]2+-CE-DAD) was developed to screen active components with the ability to chelate ferrous ions and determine the total antioxidant activity. The MAE conditions, including methanol concentration, extraction power, extraction time, and the ratio of material to liquid, were optimized by an L9(34) orthogonal experiment. Background buffer, voltage, and cartridge temperature that affect the separation of six compounds were optimized. It was found that rutin and quercetin were the main components chelating ferrous ions in Flos Sophorae Immaturus (Flos Sophorae) by the in-capillary [Fe(ferrozine)3]2+-CE-DAD. The recoveries were ranged from 95.2% to 104%. It was concluded that the MAE combined with in-capillary [Fe(ferrozine)3]2+-CE-DAD method was a simple, reliable, and efficient tool for screening active components from the complex traditional Chinese medicine samples and evaluating their ability to chelate ferrous ions.

In order to enrich and separate three coumarins (columbianetin acetate, osthole and columbianadin) from Angelicae Pubescentis Radix (APR), an efficient method was established by combining macroporous resins (MARs) with preparative high-performance liquid chromatography (PHPLC). Five different macroporous resins (D101, AB-8, DA-201, HP-20 and GDX-201) were used to assess the adsorption and desorption characteristics of three coumarins. The result demonstrated that HP-20 resin possessed the best adsorption and desorption capacities for these three coumarins. Moreover, the adsorption dynamics profiles of three coumarins were well fitted to the pseudo second order equation (R2 > 0.99) for the HP-20 resin. The adsorption process was described by the three isotherms models including Langmuir (R2 > 0.98, 0.046 ≤ RL ≤ 0.103), Freundlich (R2 > 0.99, 0.2748 ≤ 1/n ≤ 0.3103) and Dubinin Radushkevich (R2 > 0.97). The contents of columbianetin acetate, osthole and columbianadin in the product were increased 10.69-fold, 19.98-fold and 19.68-fold after enrichment, respectively. Three coumarins were further purified by PHPLC and the purities of them reached above 98%. Additionally, the anti-inflammatory effects of these three coumarins were assessed by Lipopolysaccharide (LPS)-induced RAW 264.7 cells. It was found that the production of NO and MCP-1 was obviously inhibited by three coumarins. Columbianetin acetate, osthole and columbianadin could be used as potentially natural anti-inflammatory ingredients in pharmaceutical products. It was concluded that the new method combining MARs with PHPLC was efficient and economical for enlarging scale separation and enrichment of columbianetin acetate, osthole and columbianadin with anti-inflammatory effect from the APR extract.

A sensitive and effective method was developed for clarifying the pharmacokinetic properties of six compounds (including hyperin, chlorogenic acid, neochlorogenic acid, p-coumaric acid, astragalin, and isoquercitrin) in two processed Cuscutae Semen samples by high performance liquid chromatography mass spectrometry (HPLC-MS/MS). The six compounds were separated by acetonitrile and 0.1% formic acid-water on an Agilent Eclipse plus C18 column (4.6 mm × 100 mm, 1.8 μm). All compounds were analyzed with negative ion mode in multiple reaction monitoring (MRM). The lower limits of quantification (LLOQ) of hyperin, astragalin, neochlorogenic acid, chlorogenic acid, isoquercitrin, and p-coumaric acid were 1, 0.1, 4, 0.1, 2, and 4 ng·mL−1, respectively. The validated approach was effectively used for the pharmacokinetics of six compounds of two processed Cuscutae Semen samples after oral administration to rat. The results indicated that salt processing could improve the adsorption and bioavailability of astragalin in Cuscutae Semen.

Epimedium herb is one of the most vital traditional Chinese medicines (TCMs), which is used for “nourishing the kidney and reinforcing the Yang”. In the guidance of TCM theory, Epimedium herb is usually processed with lamb oil to increase its efficacy. The contents of active ingredients in different Epimedium are significantly varied, which may derive from their different species, regions and processing methods. In this research, 13 batches of raw Epimedium collected from 6 provinces were identified. After optimization of the processing method of Epimedium, a liquid chromatography–mass spectrometry (LC–MS/MS) method for simultaneous determination of 16 compounds was established to evaluate the quality of raw and processed. Then the multivariate statistical technique was applied to compare different batches of Epimedium based on the LC–MS/MS data. As a conclusion, the herbs collected from 6 areas were ascribed to 5 species by microscopic and appearance features. Meanwhile, all of the raw and processed samples were classified by partial least squares discriminant analysis (PLS-DA) based on the 16 analyzed compounds. The comparison results indicate that processing and species both have important influences on Epimedium compositions contents.

A rapid and reliable HPLC-MS/MS method has been developed and validated for the simultaneous quantification of twelve bioactive compounds (baohuoside II, baohuoside I, sagittatoside A, sagittatoside B, magnoflorine, epimedin A, epimedin B, epimedin C, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid and icariin) in rat plasma. The collected plasma samples were prepared by protein precipitate with acetonitrile. The twelve compounds were separated on a CORTECS®C18 column (4.6 mm × 150 mm, 2.7 μm) with a gradient mobile phase system of 0.1% (v/v) formic acid and acetonitrile at a flow rate of 0.3 mL/min. All of the analytes were quantitated using electrospray ionization (ESI) in negative ion mode with selected reaction monitoring (SRM). The intra- and inter-day accuracy ranged from −5.6% to 13.0%, and the precisions of the analytes were less than 10.9%. The mean recoveries of the analytes were in the range of 60.66% to 99.77% and the matrix effect ranged from 93.08% to 119.84%. Stability studies proved that the analytes were stable under the tested conditions, with a relative standard deviation (RSD) lower than 11.7%. The developed method was successfully applied to evaluating the pharmacokinetic study of twelve bioactive compounds after oral administration of Epimedium extract in rat.