Online preconcentration and determination of five alkaloids in Yangxinshi tablet by large-volume sample stacking and micelle to solvent stacking in cyclodextrin-modified electrokinetic chromatography.

The two-step preconcentration technique consisting of large-volume sample stacking (LVSS) and micelle to solvent stacking (MSS) in cyclodextrin-modified electrokinetic chromatography (CDEKC) was developed for the analysis of five cationic alkaloids in complex Chinese herbal prescriptions. Relevant parameters affecting separation and stacking performance were optimized separately. Under the optimal LVSS-MSS-CDEKC conditions, less analysis time and organic solvent were required, and the enhancement factors of analytes ranged from 12 to 15 compared with the normal CDEKC separation mode. Further, all validation results demonstrated good applicability and multiple alkaloids (epiberberine, dehydrocorydaline, jatrorrhizine, coptisine and berberine) in Yangxinshi tablet (YXST) have been simultaneously determined. This approach presents powerful potential for the determination of multiple components in complex preparations of Chinese medicine.

Combination of large-volume sample stacking with polarity switching and micelle electrokinetic chromatography for the analysis of anion compounds in Yangxinshi tablets.

INTRODUCTION: Yangxinshi tablet (YXST) is a traditional Chinese medicine preparation characterized by its high efficacy and safety for the treatment of cardiovascular diseases. Anionic compounds have been revealed as potential active components. However, there is currently limited research regarding its quality control. OBJECTIVE: We aimed to establish a strategy for the simultaneous separation and determination of five key anionic compounds in YXST. METHOD: A sensitive and efficient analytical method was developed and applied for the simultaneous separation and determination of five key compounds in YXST using large-volume sample stacking with polarity switching and micelle electrokinetic chromatography (LVSS-PS-MEKC) coupled with diode array detection. Crucial parameters, including sample volume, applied voltage, composition and pH of the running buffer, concentration of organic modifier, and switching time of the polarity, were systematically evaluated and optimized using a single variable method to enhance separation performance. Furthermore, the impact of cyclodextrin and sodium dodecyl sulfate as electrolyte modifiers was also investigated. RESULTS: Under the optimal conditions, baseline separation of the five compounds (daidzein, puerarin, glycyrrhiztinic acid, chlorogenic acid, and salvianolic acid B) was achieved within 20 min. In comparison to the conventional MEKC mode, the constructed LVSS-PS-MEKC method exhibited a more than sixfold increase in the enrichment factor. The method was validated in terms of linearity, precision, accuracy, 24 h stability, and recovery and successfully applied to analyze YXST samples. CONCLUSION: A sensitive strategy was developed for the simultaneous separation and determination of five key anionic components in YXST, offering a robust and efficient strategy for pharmaceutical analysis.

Quantification and discovery of quality markers from Toddalia asiatica by UHPLC-MS/MS coupled with chemometrics.

INTRODUCTION: Toddalia asiatica (TA) is a classical traditional Chinese medicine used to treat rheumatoid arthritis and contusions. However, research regarding TA quality control is currently limited. OBJECTIVE: We aimed to establish a strategy for identifying quality markers that can be used for the evaluation of the quality of TA. METHOD: A rapid and efficient ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitative determination of 19 compounds in TA from different regions. Then, the extraction process of TA was successively optimized by single-factor optimization and response surface methodology. Moreover, chemometrics was employed to confirm the correlation between quality and target compounds. RESULTS: Utilizing the UHPLC-MS/MS method, separation of the 19 bioactive compounds was achieved within 14 min. The method was validated in terms of linearity (r2 > 0.9982), precision (0.08%-3.70%), repeatability (0.50%-2.54%), stability (2.26%-5.46%), and recovery (95.8%-113%). The optimal extraction process (extraction solvent, 65% ethanol aqueous solution; solid-liquid ratio, 1:20; extraction time, 25 min) was determined with the total content of 19 bioactive compounds as indicator. Significant disparities were observed in the contents of target compounds across different batches of TA. Besides, all samples could be categorized into two distinct groups, and magnoflorine, (-)-lyoniresinol, nitidine chloride, norbraylin, skimmianine, and decarine were identified as quality markers. CONCLUSION: In the present study, we developed a strategy to improve the quality control of TA. In consideration of the pharmacodynamic activity and statistical differences, six compounds are proposed as quality markers for TA.

A strategy of accuracy quantification by extending the concentration monitoring coverage based on online double collision energy of ultra-high performance liquid chromatography tandem mass spectrometry: The pharmacokinetics of Toddalia asiatica as a case study.

To facilitate the safety, efficacy and rationality of clinical application of traditional Chinese medicines (TCMs), pharmacokinetic research played an indispensable role. The key challenge during pharmacokinetic investigation lied at the substantial fluctuation of compound concentrations in the plasma over the course of absorption. Taking the pharmacokinetics of six compounds after administration of Toddalia asiatica (TA) as an example, an efficient strategy was established by introducing the online double collision energy (ODCE) into the quantification process applying ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). During the analytical program, double collision energy (DCE) was optimized to establish the dual calibration curve (DCC) with large concentration monitoring coverage (CMC) for meeting the wide content range of certain target compounds. Method validation test was performed in terms of linearity, precision, sensitivity, matrix effect, recovery, etc. The results displayed that the CMC of todarolactone with high exposure in plasma was extended from 1.25-2,500 ng/mL to 1.25-125,000 ng/mL. Furthermore, a rapid UHPLC-MS/MS method integrated with ODCE was successfully applied to the determination of six compounds in rat plasma, revealing an extremely high plasma concentration of todarolactone (16,662 ng/mL). This strategy could expand the range of quantification while retaining extraordinary sensitivity. Consequently, it could be a fit-for-purpose strategy to quantify compounds over a wide concentration range for in vivo process monitoring.

A comprehensive review of the botany, ethnopharmacology, phytochemistry, pharmacology, quality control and other applications of Ligustici Rhizoma et Radix.

ETHNOPHARMACOLOGICAL RELEVANCE: Ligusticum sinense Oliv. and L. jeholense Nakai et Kitag. are globally recognized as medicinal botanical species, specifically the rhizomes and roots. These plant parts are collectively referred to as Ligustici Rhizoma et Radix (LReR), which is recorded in the Pharmacopoeia of the People's Republic of China (Ch. P). LReR enjoys widespread recognition in many countries such as China, Russia, Vietnam, and Korea. It is an herbal remedy traditionally employed for dispelling wind and cold, eliminating dampness, and alleviating pain. Numerous bioactive compounds have been successfully isolated and identified, displaying a diverse array of pharmacological activities and medicinal value. THE AIM OF THE REVIEW: This review aims to primarily center on the botanical aspects, ethnopharmacology, phytochemistry, pharmacology, toxicity, quality control, and other applications of LReR to furnish a comprehensive and multidimensional foundation for future exploration and utilization. MATERIALS AND METHODS: Relevant information about LReR was acquired from ancient books, doctoral and master's dissertations, Google Scholar, Web of Science, PubMed, China National Knowledge Infrastructure (CNKI), ScienceDirect, classical literature, and clinical reports. Several electronic databases were also incorporated. RESULTS: In traditional usage, LReR had been traditionally employed for the treatment of anemofrigid headaches, colds, and joint pain. It possessed therapeutic properties for facial skin disorders, thereby facilitating skin regeneration. It has been subjected to comprehensive chemical analysis, resulting in the identification and isolation of 190 compounds, including phthalides, phenylpropanoids, flavonoids, phenolic acids, triterpenes, steroids, volatile oil, fatty acids, and other constituents. The pharmacological activities have been in-depth explored through modern in vivo and in vitro studies, confirming its anti-inflammatory, analgesic, and anti-melanin effects. Furthermore, it exhibited pharmacological activities such as antioxidant, anticancer, antibacterial, and vasodilatory properties. This study provides a basic to contribute to the advancement of research, medicinal applications and product development related to LReR. CONCLUSIONS: Considering its traditional and contemporary applications, phytochemical composition, and pharmacological properties, LReR was regarded as a valuable botanical resource for pharmaceutical and pest control purposes. While certain constituents had demonstrated diverse pharmacological activities and application potential, further elucidation was required to fully understand their specific actions and underlying mechanisms. Hence, there was a need to conduct additional investigations to uncover its material foundation and mode of action.

A global profiling strategy for identification of the total constituents in Chinese herbal medicine based on online comprehensive two-dimensional liquid chromatography-quadrupole time-of-flight mass spectrometry combined with intelligentized chemical classification guidance.

A comprehensive strategy for effective identification of total constituents in Chinese patent medicine has been advanced applying full scan-preferred parent ions capture-static and active exclusion (FS-PIC-SAE) acquisition coupled with intelligent deep-learning supported mass defect filter (MDF) process, with Naoxintong capsule (NXT) as a case. Online comprehensive two-dimensional liquid chromatography (2DLC) coupled with Q-TOF-MS/MS system was established for obtaining the excellent separation and detection performance of total components, which could exhibit excellent peak capacity with 1052 and orthogonality with 0.69. In addition, a total of 901 unknown compounds could be classified into nine chemical classes rapidly and effectively, based on the intelligent deep-learning algorithm supported MDF model with 96.4% accuracy. Consequently, 276 compounds were successfully identified from NXT, especially including 44 flavonoids, 27 phenolic acids, 25 fatty acids, 17 saponins, 21 phthalocyanines, 20 triterpenes, 10 monoterpenes, 13 diterpenoid ketones, 14 amino acids, and others. It is concluded that the proposed program is an effective and practical strategy enabling the in-depth chemical profiling of complex herbal and biological samples.

Simultaneous Separation and Determination of Nine Active Ingredients in Sanyetangzhiqing by Cyclodextrin-Modified Micellar Electrokinetic Capillary Electrophoresis-Diode Array Detector.

A simple and sensitive strategy using cyclodextrin-modified micellar electrokinetic chromatography with diode array detector was developed and applied for the simultaneous separation and determination of nine components in Sanyetangzhiqing (SYTZQ), a hypoglycemic and hypolipidemic agent. Several important parameters affecting separation performance were evaluated and optimized using single variable methods. Under the optimal conditions, baseline separation of the nine components, including four flavonoids (hyperoside, isoquercitrin, quercetin-3-O-glucuronoside, and astragalin), four phenolic acids (chlorogenic acid, rosmarinic acid, salvianolic acid B, and lithospermic acid), and a monoterpenoids (paeoniflorin), were achieved in less than 16 min. The correlation coefficients of the calibration curves were over 0.9996 for all the analytes. Intraday and interday precisions ranged from 0.4% to 4.8% and 1.7% to 5.0%, respectively. Recoveries of analytes varied from 95.3% to 105%. Validation results as well as the application to analyse SYTZQ samples demonstrated the applicability of the proposed method and thus provided an effective tool for the quality control of SYTZQ. Moreover, with the advantages of short time consuming, low energy consumption, high efficiency, and low cost, this method has laid a foundation for the determination and quality evaluation of multicomponents in Chinese herbal compounds.

Simultaneous determination of multiple components in rat plasma by UHPLC-sMRM for pharmacokinetic studies after oral administration of Qingjin Yiqi Granules.

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.

An efficient method for qualitation and quantitation of multi-components of the herbal medicine Qingjin Yiqi Granules.

Qingjin Yiqi Granules (QJYQ) is a Traditional Chinese Medicines (TCMs) prescription for the patients with post-COVID-19 condition. It is essential to carry out the quality evaluation of QJYQ. A comprehensive investigation was conducted by establishing deep-learning assisted mass defect filter (deep-learning MDF) mode for qualitative analysis, ultra-high performance liquid chromatography and scheduled multiple reaction monitoring method (UHPLC-sMRM) for precise quantitation to evaluate the quality of QJYQ. Firstly, a deep-learning MDF was used to classify and characterize the whole phytochemical components of QJYQ based on the mass spectrum (MS) data of ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Secondly, the highly sensitive UHPLC-sMRM data-acquisition method was established to quantify the multi-ingredients of QJYQ. Totally, nine major types of phytochemical compounds in QJYQ were intelligently classified and 163 phytochemicals were initially identified. Furthermore, fifty components were rapidly quantified. The comprehensive evaluation strategy established in this study would provide an effective tool for accurately evaluating the quality of QJYQ as a whole.

A comprehensive review of the botany, ethnopharmacology, phytochemistry, pharmacology, toxicity and quality control of Perillae Fructus.

ETHNOPHARMACOLOGICAL RELEVANCE: Perilla frutescens (Linnaeus) Britton, Mem. Torrey Bot. Club 5: 277. 1894., is famous as a worldwide plant with multiple medical parts, including leaves, stems, fruits, etc. Perillae Fructus, the desiccative ripe fruit of P. frutescens, is locally called Zisuzi in Chinese Pharmacopoeia. It is a popularly used herb for relieving cough and asthma, dissipating phlegm and treating constipation in some Asian countries, such as China, Japan, India, South Korea, etc. Various chemical compounds were isolated and identified from Perillae Fructus. THE AIM OF THE REVIEW: This review aims to summarize the botany, ethnopharmacological applications, phytochemistry, pharmacology, toxicity and quality control of Perillae Fructus to provide scientific evidence for development and utilization Perillae Fructus. MATERIALS AND METHODS: Relevant information about Perillae Fructus was collected from ScienceDirect, PubMed, Web of science, CNKI, WanFang data, ancient classics and clinical reports. Some electronic databases were also retrieved. RESULTS: Perillae Fructus was exerted to treat cough and asthma in traditional application. It also had the effect on moistening intestine to relieve constipation for tremendous lipid substances. Up to now, 193 compounds have been isolated and identified from Perillae Fructus, mainly including fatty acids, flavonoids, phenolic acids, phytosterols, triterpenoids and volatile oils. As for its pharmacological activities, prevalent traditional applications of Perillae Fructus have been supported by modern pharmacological experiments in vivo or in vitro, such as anti-inflammatory and anti-oxidant effects. Besides, Perillae Fructus also has hypolipidemic, anti-tumor, antibacterial effects, etc. This review will provide a scientific basis for further studies and rational applications of Perillae Fructus in the future. CONCLUSIONS: According to its traditional applications, phytochemicals and pharmacological activities, Perillae Fructus was regarded as a valuable herb for application in medicine and food fields. Although some ingredients have been confirmed to have multiple pharmacological activities, their mechanisms of action are still unclear. Further studies on the material basis and mechanism of action are clearly warranted.

A green ultrasonic-assisted micellar extraction coupled with ultra-high performance liquid chromatography with photodiode array method for quantitative analysis of active ingredients in Yangxinshi Tablet.

An efficient and green ultrasonic-assisted micellar extraction method coupled with ultra-high performance liquid chromatography with photodiode array detection (UHPLC-PDA) was developed for the multi-ingredients quantitative analysis of Yangxinshi Tablet (YXST). The active ingredients were extracted from YXST using trehalose lipid biosurfactant solution as an environmentally friendly extraction solution. The response surface methodology (RSM) based on Box-Behnken design (BBD) was utilized to seek for the optimum extraction conditions of target analytes. When the concentration of trehalose lipid solution was 7 mg/mL, the liquid to solid was 125:1 (mL:g) and the extraction time was 40 min, the total extraction yield of eleven compounds (including puerarin, daidzin, ferulic acid, calycosin-7-O-ß-D-glucoside, tetrahydropalmatine, coptisine, epiberberine, jatrorrhizine, berberine, palmatine chloride and icariin) obtained the maximum value. The relative standard deviations (RSD) of intra-day and inter-day precision were all less than 5.0%. The recoveries of all analytes were in the range of 95.1%-104% with the RSDs were all below 3.0%. Consequently, the ultrasonic-assisted micellar extraction coupled with UHPLC-PDA method could be successfully and efficiently applied to the extraction and quantitative analysis of target components in YXST.

A comprehensive strategy integrating metabolomics with multiple chemometric for discovery of function related active markers for assessment of foodstuffs: A case of hawthorn (Crataegus cuneata) fruits.

There is a growing demand for functional foodstuffs naturally rich in nutrients and bioactive compounds associated with putative health benefits. This study followed a strategy integrating metabolomic with chemometric to identify quality markers that can be used to distinguish function related bioactivity of fruits, using hawthorn (Crataegus cuneata) fruit as an example. The integrated strategy was reduced to characterization of metabolic processes and discovered potential bioactive ingredients that were further investigated for functionalities. This comprehensive process provided guidance for discovery of function-related active markers for assessment of foodstuffs.

An integration strategy combined progressive multivariate statistics with anticoagulant activity evaluation for screening anticoagulant quality markers in Chinese patent medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: The cardiovascular and cerebrovascular diseases affect human health globally. Naoxintong capsules (NXTs), a famous Chinese Patent Medicine, has been especially applied to treat cerebral infarction and coronary heart disease in clinical practice. The anticoagulant activity of this prescription plays an important role in this course of treatment. AIM OF THE STUDY: Thrombin and factor Xa (FXa) are two key targets considering the anticoagulant activity. The purpose of this investigation is to screen the quanlity markers as key thrombin and FXa inhibitors for the anticoagulant activity oriented quality control of Chinese patent medicine. MATERIALS AND METHODS: Simple multi-polar solvent extraction processes using various proportions of solvents were conducted and their thrombin/FXa inhibitory activities were evaluated in vitro. Bivariate correlation analysis (BCA), grey correlation analysis (GCA), and orthogonal partial least squares discriminate analysis (OPLS-DA) were adopted for screening the potential active markers related to the anticoagulant activity. The chemical structures of these active compounds were identified by UHPLC-Q-TOF-MS/MS and their thrombin/FXa inhibitory activity was determined. The molecular docking technology was applied to explore the interaction between the compounds and targets. The contribution of these anticoagulant active ingredients in NXT was also investigated. Last but not the least, the contents of these markers in NXT were determined by liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (HPLC-ESI-MS/MS) method. RESULTS: The results showed that the NXT extract exhibited great activity against thrombin and FXa, especially extracted by 75% methanol (v/v). Six marker compounds with potential anticoagulant activity were screened out. Therein, four of the active compounds owing thrombin inhibitory activity (paeoniflorin, lithospermic acid, salvianolic acid B, Z-ligustilide) and five of the active compounds owing FXa inhibitory activity (3,5-dicaffeoylquinic acid, rosmarinic acid, lithospermic acid, salvianolic acid B and Z-ligustilide). In addition, these active compounds accounted for a large proportion of thrombin/FXa inhibitory activity of NXTs. The binding energy also showed the strong interaction formed by close connection of the compounds to the residues of targets. CONCLUSIONS: The proposed integrated stategy could be an efficient strategy to screen potential thrombin/FXa inhibitors for the bioactivity related quanlity control of Chinese patent medicine.

Immunosuppressive effect of Columbianadin on maturation, migration, allogenic T cell stimulation and phagocytosis capacity of TNF-α induced dendritic cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelicae pubescentis radix (APR) has a long history in the treatment of rheumatoid arthritis (RA) in China. It has the effects of dispelling wind to eliminate dampness, removing arthralgia and stopping pain in the Chinese Pharmacopeia, but its mechanisms was unclear. Columbianadin (CBN) was one of the main bioactive compounds of APR, and has many pharmacological effects. But the immunosuppressive effect of CBN on DCs and the potential mechanism needed to be explored. AIM OF THE STUDY: The study was aimed to clarify the immunosuppressive effect of CBN on maturation, migration, allogenic T cell stimulation and phagocytosis capacity of TNF-α induced DCs. MATERIALS AND METHODS: Bone marrow-derived DCs were obtained and cultured from C57BL/6 mice in accordance with protocol. The phenotypic study (CD11c, CD40, CD80, CD86 and MHC Ⅱ) were measured by flow cytometry. FITC-dextran were uptaked by DCs and the change of endocytosis activity were mediated by acquired mannose receptor. Transwell chambers were used to detect the migration ability of DCs. Mixed leukocyte reaction (MLR) assay was used to detect the allostimulatory ability of CBN on TNF-α stimulated DCs. The secretion of cytokines and chemokines was measured by ELISA Kit. TLRs gene and MAPKs/NF-κB protein expression were checked by qRT-PCR and Western blot. RESULTS: CBN inhibited the maturation of TNF-α-induced DCs while maintaining phagocytosis capabilities. Additionally, CBN inhibited the migration of TNF-α stimulated DCs, which related to reduce the production of chemokines (MCP-1, MIP-1α). Notably, CBN could suppress the proliferation of CD4+T cells by inhibiting DCs maturation, and decrease the proinflammatory cytokines IL-6 production. Furthermore, CBN inhibited mRNA expression of TLR2, TLR7 and TLR9 in TNF-α-activated DCs. Meanwhile, the phosphorylation of p38, JNK1/2 and NF-κB protein were significantly inhibited in CBN treated DCs. CONCLUSIONS: These findings provided novel insights into the pharmacological activity of CBN. They also indicated that inhibition DCs maturation owning to the immunosuppressive effect of CBN. CBN was expected as a potential immunosuppressant and TLRs/MAPKs/NF-κB pathway may be an important mechanism for CBN's immunosuppressive activity.

Traditional herbal medicine: Therapeutic potential in rheumatoid arthritis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease influenced by diverse endogenous and exogenous factors. It is characterized by cartilage and bone destruction. The current conventional allopathic therapy is expensive and carries adverse side effects. Recently, there were some ethnopharmacological studies on RA including anti-RA effects and therapeutic targets of distinct dosage forms of traditional herbal medicines (THMs). AIM OF THE REVIEW: This review provides a brief overview of the current understanding of the potential pharmacological mechanisms of THMs (active constituents, extracts and prescriptions) in RA. This study is intended to provide comprehensive information and reference for exploring new therapeutic strategies of THMs in the RA treatment. MATERIALS AND METHODS: This review captured scientific literatures invivo and vitro experiments on effects of anti-RA THMs published between 2016 and 2021 from journals and electronic databases (e.g. PubMed, Elsevier, Science Direct, Web of Science and Google Scholar). Relevant literatures were searched and analyzed by using keywords such as 'rheumatoid arthritis AND traditional herbal medicines', 'rheumatoid arthritis AND immune cells', 'rheumatoid arthritis AND inflammation', 'rheumatoid arthritis AND miRNA', 'rheumatoid arthritis AND Angiogenesis', 'rheumatoid arthritis AND oxidative stress', 'rheumatoid arthritis AND osteoclasts', 'rheumatoid arthritis AND CIA model', 'rheumatoid arthritis AND AA model' AND 'rheumatoid arthritis herbal prescription'. RESULTS: Experiments in vitro and in vivo jointly demonstrated the potential of THMs in the RA treatment. There are plentiful therapeutic targets in RA. THMs and active ingredients could alleviate RA symptoms through different therapeutic targets, such as immunoregulation, inflammation, fibroblast-like synoviocytes (FLSs), microRNAs (miRNAs), angiogenesis, oxidative stress, osteoclasts and multiple targets interaction. Anti-RA THMs, active ingredients and prescriptions through corresponding therapeutic targets were summarized and classified. CONCLUSIONS: Flavonoids, phenolic acids, alkaloids and triterpenes of THMs are identified as the main components to ameliorate RA. Regulation of different and multiple related therapeutic targets by THMs and their active ingredients were associated with greater therapeutic benefits, among which inflammation is the main therapeutic target. Nonetheless, further studies are required to unravel the complexities and in-depth mechanisms of THMs in alleviating RA.

Biosurfactant trehalose lipid-enhanced ultrasound-assisted micellar extraction and determination of the main antioxidant compounds from functional plant tea.

Hydrosoluble trehalose lipid (a biosurfactant) was employed for the first time as a green extraction solution to extract the main antioxidant compounds (geniposidic acid, chlorogenic acid, caffeic acid, and rutin) from functional plant tea (Eucommia ulmoides leaves). Single-factor tests and response surface methodology were employed to optimize the extraction conditions for ultrasound-assisted micellar extraction combined with ultra-high-performance liquid chromatography in succession. A Box-Behnken design (three-level, three-factorial) was used to determine the effects of extraction solvent concentration (1-5 mg/mL), extraction solvent volume (5-15 mL), and extraction time (20-40 min) at a uniform ultrasonic power and temperature. In consequence, the best analyte extraction yields could be attained when the trehalose lipid solution concentration was prepared at 3 mg/mL, the trehalose lipid solution volume was 10 mL and the extraction time was set to 35 min. In addition, the recoveries of the antioxidants from Eucommia ulmoides leaves analyzed by this analytical method ranged from 98.2 to 102%. These results indicated that biosurfactant-enhanced ultrasound-assisted micellar extraction coupled with a simple ultra-high-performance liquid chromatography method could be effectively applied in the extraction and analysis of antioxidants from Eucommia ulmoides leaf samples.

Determination of antioxidant ingredients in Mori Fructus employing ionic liquid-assisted miniaturized matrix solid-phase dispersion extraction via ultra-performance liquid chromatography.

A simple miniaturized matrix solid-phase dispersion extraction was developed to analyze multiple antioxidant ingredients (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, and astragalin) in Mori Fructus by ultra performance liquid chromatography coupled with an ultraviolet detector. 1-ethyl-3-methylimidazolium bromide was employed in the elution process as eco-friendly elution solvent. The effects of some factors on the extraction efficiency of antioxidants in Mori Fructus were optimized in detail, such as the type of adsorbent, the type of elution solvent, the concentration and volume of elution solvent, sample to adsorbent ratio, and the grinding time. The intra-day and inter-day precisions (RSD) were both below 5.0% and the recoveries of all analytes ranged from 93.5% to 98.3%. Compared with the traditional method, it was efficient, simple and environment-friendly for extracting the analytes by ionic liquid assisted trace ß-CD matrix solid-phase dispersion extraction method, due to shorter extraction time, less reagent and less sample consumption. The developed method was successfully used for extracting and determining the active compounds in Mori Fructus sample. PRACTICAL APPLICATIONS: The antioxidant ingredients play important roles in Mori Fructus because of their main pharmacological activities. Nowadays, a few analytical methods could be applied for extracting and analyzing these target compounds. But these methods not only required much extracting and analyzing time, but also need a great deal of organic reagent, which were not environmentally friendly. Thus, a green and simple miniaturized matrix solid phase dispersion extraction method was proposed for the analysis of these antioxidants in Mori Fructus, which was efficient to be employed for evaluating the quality of Traditional Chinese Medicine.

Quantification of eight active ingredients in crude and processed radix polygoni multiflori applying miniaturized matrix solid-phase dispersion microextraction followed by UHPLC.

A rapid, efficient, and green sample preparation method has been developed to extract eight active ingredients (gallic acid, catechins, epicatechin, polydatin, 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucoside, resveratrol, emodin, and physcion) in radix polygoni multiflori by miniaturized matrix solid-phase dispersion microextraction. Simple and sensitive ultra high performance liquid chromatography combined with ultraviolet detection has been applied to analyze the multiple compounds. The best results were obtained by adding 25 mg sample into 25 mg adsorbent and grinding for 2 min with disorganized silica as adsorbent and 1 mL 150 mM 1-dodecyl-3-methylimidazolium bromide as a green eluting solvent. Good linearity (r2  > 0.998) for each analyte was obtained by this method. The intra-day and inter-day precision (RSD) were both below 5.31%, and the recoveries of the analytes ranged from 93.3 to 100.0%. This simple miniaturized matrix solid-phase dispersion microextraction method for analyzing the compounds in radix polygoni multiflori needs a short time and requires little sample and reagent. Thus, this method is far more eco-friendly and efficient than traditional extraction methods (reflux and ultrasound-assisted extraction). The present investigation provided a promising method for the fast preparation and discrimination of chemical differences in crude and processed radix polygoni multiflori.

Non-ionic detergent Triton X-114 Based vortex- synchronized matrix solid-phase dispersion method for the simultaneous determination of six compounds with various polarities from Forsythiae Fructus by ultra high-performance liquid chromatography.

A simple nonionic detergent - based vortex- synchronized matrix solid-phase dispersion (ND-VSMSPD) method was developed to extract bioactive compounds in Forsythiae Fructus coupled with ultra high-performance liquid chromatography (UHPLC). Nonionic detergent Triton 114 was firstly used as a green elution reagent in vortex- synchronized MSPD procedure. The optimum parameters were investigated to attain the best results, including Florisil as sorbent, 2mL 10% (v/v) nonionic detergent Triton X-114 as the elution reagent, 1:1 of sample/sorbent ratio, grinding for 3min, and whirling for 2min. The recoveries of the six compounds in Forsythiae Fructus were in the range of 95-104% (RSD <4.6%) and the method displayed a good linearity within the range of 0.08-20µgmL-1 for caffeic acid, 0.6-150µgmL-1 for forsythoside A, 0.4-100µgmL-1 phillyrin, 0.2-50µgmL-1 for quercetin, isorhamnetin and arctigenin (r≥0.999). It was proved that the extraction yields of almost all compounds attained by the established vortex- synchronized MSPD, which required lower sample, reagent and time, were higher than the normal MSPD and the traditional ultrasonic-assisted extraction. Consequently, this developed vortex- synchronized MSPD coupled with simple UHPLC method could be efficiently applies to extract and analyze the target compounds in real Forsythiae Fructus samples.