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.

Xuebijing injection inhibited neutrophil extracellular traps to reverse lung injury in sepsis mice via reducing Gasdermin D.

The mortality of sepsis and septic shock remains high worldwide. Neutrophil extracellular traps (NETs) release is a major cause of organ failure and mortality in sepsis. Targeting Gasdermin D (GSDMD) can restrain NETs formation, which is promising for sepsis management. However, no medicine is identified without severe safety concerns for this purpose. Xuebijing injection (XBJ) has been demonstrated to alleviate the clinical symptoms of COVID-19 and sepsis patients, but there are not enough animal studies to reveal its mechanisms in depth. Therefore, we wondered whether XBJ relieved pulmonary damage in sepsis by suppressing NETs formation and adopted a clinically relevant polymicrobial infection model to test this hypothesis. Firstly, XBJ effectively reversed lung injury caused by sepsis and restrained neutrophils recruitment to lung by down-regulating proinflammatory chemokines, such as CSF-3, CXCL-2, and CXCR-2. Strikingly, we found that XBJ significantly reduced the expressions of NETs component proteins, including citrullinated histone H3 (CitH3), myeloperoxidase (MPO), and neutrophil elastase (NE). GSDMD contributes to the production of NETs in sepsis. Notably, XBJ exhibited a reduced effect on the expressions of GSDMD and its upstream regulators. Besides, we also revealed that XBJ reversed NETs formation by inhibiting the expressions of GSDMD-related genes. Collectively, we demonstrated XBJ protected against sepsis-induced lung injury by reversing GSDMD-related pathway to inhibit NETs formation.

Stereoselective enrichment and determination of citalopram enantiomers by cation-selective exhaustive injection and sweeping coupled with cyclodextrin modified electrokinetic chromatography.

An highly sensitive, rapid and enantioselective method was developed for the enantioseparation and determination of citalopram enantiomers by cation selective exhaustive injection-sweeping-cyclodextrin modified electrokinetic chromatography (CSEI-sweeping-CDEKC). The optimized conditions were: 50 mM pH 3.0 phosphate solution with 25 mg·mL-1 S-ß-CD used as background buffer, 50 mbar 300 s hydrodynamical injection of 150 mM pH 3.0 NaH2PO4 buffer followed with 5 s water plug, 10 kV 600 s electrokinetic sample injection, -20 kV CDEKC run. Under the optimized conditions, the resolution was Rs=8.04, the enrichment factor as up to 2163 folds, the LOD values were: 3.6 ng·mL-1 for R-citalopram, 4.1 ng·mL-1 for S-citalopram, and 3 ng·mL-1 for both enantiomers in plasma samples. This new method showed good precision, repeatability and stability, which had been successfully applied to the impurity inspection of escitalopram oxalate and the stereoselective pharmacokinetic study of citalopram enantiomers.

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.

A review of the ethnopharmacology, phytochemistry and pharmacology of Cynanchumatratum.

ETHNOPHARMACOLOGICAL RELEVANCE: The dried roots and rhizomes of Cynanchum atratum Bunge is named 'Baiwei' according to traditional Chinese medicine theory. It is also named Cynanchi atrati Radix in Chinese Pharmacopoeia. Cynanchi atrati Radix is famous for its medicinal value of clearing away heat, relieving drenching, detoxifying and treating abscesses. It was commonly used in some Asian countries for the treatment of fever, vasoconstrictive syncope, lymphangitis and other diseases, obviously due to the effect of C21 steroidal glycosides. THE AIM OF THE REVIEW: The review concentrates on the botany, ethnopharmacology, phytochemistry, pharmacology and toxicology of Cynanchum atratum. We also discuss expectations for prospective research and implementation of this herb. MATERIALS AND METHODS: Relevant information about C. atratum was gained from ancient books and records, Doctoral and master's Theses, Science Direct, Pubmed, Wiley, CNKI, WanFang DATA, Google Scholar and other domestic and foreign literature. Some electronic databases have been included. RESULTS: As a member of the Apocynaceae family, C. atratum possesses its up-and-coming biological characteristics. It is widely reported for treating of postpartum fatigue, vomiting, urine drops, nephritis, urinary tract infection, edema, bronchitis and rheumatic low back pain. By now, over 100 compounds have been identified from C. atratum, including C21 steroidal glycosides, acetophenones, alkaloids, volatile oil and other ingredients. Activities such as anti-inflammatory, anti-tumor, anti-virus, antibacterial, anti-forgetful and others have been corroborated in vivo and in vitro. In addition, many of the active ingredients, such as Cynatratoside A, Cynanversicoside A, B, D, G, p-hydroxyacetophenone, 2,4-dihydroxyacetophenone and some volatile oils have been used as quality markers. CONCLUSION: All kinds of research conducted on C.atratum, especially in field of ethnopharmacological use, phytochemicals and pharmacology have been reviewed. The herb has been used over the years in treating nephritis, urinary tract infection, bronchitis and rheumatic lumbocrural pain. Many studies have been carried out to identify compounds that play a leading role in drug activity. However, the mechanism of drug therapy remains unclear. The evidence used to prove the quality standard of medicinal materials is obviously inadequate. Besides, safety evaluation is necessary for clinical medication. Similarly, the separation of steroidal saponins and the development of new drugs will also need further discussion.

Binary Eluent Based Vortex-Assisted Matrix Solid-Phase Dispersion for the Extraction and Determination of Multicomponent from Musk by Gas Chromatography-Mass Spectrometry.

A green, flexible, and effective strategy was proposed to quantify four target compounds (muscone, ethyl palmitate, ethyl oleate, and ethylparaben) from musk by binary eluent based vortex-assisted matrix solid-phase dispersion (MSPD) extraction coupled with GC/MS. Single-factor tests and orthogonal design were employed to optimize the MSPD parameters. In addition, the binary eluent system, methanol, and ethyl acetate 3 : 7 (v/v) were used to extract the target analytes. Finally, C 18 was applied as the easily available dispersant and the sample powder was ground for 2 min. Thereafter, the mixture was rapidly extracted with the binary eluents under whirling for 3 min. Eventually, the analysis of the samples was completed within 12 min by GC/MS. All correlation coefficients (r) of four targets were more than 0.9991. The recoveries of all target compounds ranged from 92.8% to 101% while their RSDs were less than 6.94%. There was no significant matrix interference for the analysis. Thus, the combination of vortex-assisted MSPD with GC/MS was considered as a novel, rapid, and environmentally friendly quantitative approach for musk samples.

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.

The network pharmacology integrated with pharmacokinetics to clarify the pharmacological mechanism of absorbed components from Viticis fructus extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Viticis fructus (VF) has been widely used in alleviating the swelling and pain, owning to its pharmacologically active components including agnuside, 10-O-vanilloylaucubin, luteolin and casticin. AIM OF THE STUDY: The pharmacokinetic profiles of the absorbed components from aqueous and ethanolic extracts of VF in rat plasma were performed, and explored the molecular mechanisms of absorbed components via network pharmacology. MATERIALS AND METHODS: Ultra-performance liquid chromatography-tandem mass spectroscopy (UHPLC-MS/MS) was employed to identify the absorbed components from rat plasma. Liquid-liquid extraction with ethyl acetate was used to purify the plasma samples. Plasma pharmacokinetics parameters of the components absorbed were analyzed after oral administration of both extracts. Network pharmacology was used to predict the biological functions and potential signaling pathways of VF. The anti-cancer effects of VF extract and absorbed components have been confirmed by in vitro experiments. RESULTS: The method was very sensitive with lower limit of quantification (LLOQ) of 1.0, 2.5, 0.2 and 0.5 ng/mL for agnuside, 10-O-vanilloylaucubin, luteolin and casticin, respectively. With the exception of 10-O-vanilloylaucubin which was not detected in the ethanolic extract of VF, all other components were detected in both extracts in plasma. The pharmacokinetic parameters of the four components from rat plasma were significantly different between the two extracts. According to the results of network pharmacology, the absorption components of VF are enriched in 32 key pathways, and 15 pathways are related to cancer. Ultimately, the anti-cancer effects, as well as the signaling pathways of VF ethanolic extract and absorbed components were verified by in vitro experiments. CONCLUSION: The optimized, sensitive and validated UHPLC-MS/MS method was successfully applied for the plasma pharmacokinetics comparison analysis of the two VF extracts. The combination of network pharmacology and pharmacokinetics provides a useful method to elucidate the biological effects and molecular mechanism of the absorbed components of VF.

A strategy for intelligent chemical profiling-guided precise quantitation of multi-components in traditional Chinese medicine formulae-QiangHuoShengShi decoction.

Due to the tremendous clinical value, more and more Traditional Chinese Medicines (TCMs) and their formulae are attracted by world's attention. QiangHuoShengShi (QHSS) decoction is one of classic TCM formulae, which is clinically used for treating various rheumatic diseases. However, the phytochemical constituents of QHSS have rarely been reported. A simple, intelligent, and comprehensive strategy was developed to characterize the phytochemical-fingerprint and quantify the chemical-markers for precise quality evaluation of QHSS. Firstly, a new deep-learning assisted mass defect filter (MDF) method was built for rapid and accurate classification of mass spectrum (MS) ions acquired by ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Subsequently, herb species-specific chemical-category and characteristic identification were used for further characterization of multi-components. As the result, seven major types of compounds in QHSS were intelligently differentiated and 183 phytochemical compounds were tentatively identified. Finally, a sensitive scheduled multiple reaction monitoring (sMRM) detection method was applied to precisely quantify 37 target analytes in QHSS decoction. This integrated strategy would provide an alternative method for chemical-material basis study of more herbal medicine or natural products.

Corrigendum to "Monitoring unbound warfarin in drug combination therapy by pharmacokinetics and fluorospectrometry" [Chinese Herbal Medicines 11 (2019) 92-97].

[This corrects the article DOI: 10.1016/j.chmed.2018.10.002.].

Buyang Huanwu Decoction ameliorates atherosclerosis by regulating TGF-ß/Smad2 pathway to promote the differentiation of regulatory T cells.

ETHNOPHARMACOLOGICAL RELEVANCE: As a classic prescription of Traditional Chinese Medicine in Correction on the Errors of Medical Works, Buyang Huanwu Decoction (BYHWD) has a good curative effect on prevention of atherosclerosis (AS). AIM OF THE STUDY: This study aims to elucidate the anti-atherosclerosis mechanism of BYHWD, which may promote the differentiation of regulatory T cells by regulating the TGF-ß/Smad2 pathway. MATERIALS AND METHODS: ApoE-/- mice were fed a high-fat diet for 12 weeks, then drugs group were given BYHWD with intragastric administration once a day for 4 weeks. The effect of BYHWD on lipid content in peripheral blood and plaque was evaluated by blood lipid test and oil red O staining. The number of Tregs in peripheral blood was tested by flow cytometry, and that in the spleen was evaluated by immunohistochemistry methods. Gene and protein expression relating with Tregs differentiation pathway in mice were checked by RT-PCR and Western blot experiments. CD4+T cells were isolated and interfered by BYHWD drug-loaded serum. The proportion of Tregs was evaluated by flow cytometry. The chemical compositions of BYHWD and rat drug-loaded serum were analyzed by ultra-high performance liquid chromatograph and liquid chromatography-tandem mass spectrometry. RESULTS: BYHWD significantly reduced plaque area and cholesterol accumulation, increased the number of Tregs in spleen and peripheral blood of ApoE-/- AS mice, raised the proportion of Tregs in CD4+T cells, and regulated the levels of inflammatory factors. It also increased the TGF-ß and Smad2 mRNA and protein levels relating with Tregs differentiation pathway in vivo. The mRNA levels of Foxp3/TGF-ß/Smad2 were enhanced via BYHWD in vitro. CONCLUSIONS: BYHWD regulates TGF-ß/Smad2 signaling pathway to promotes the peripheral differentiation of Tregs, increases the number of Tregs, restores the immune balance between CD4+T cells, regulates lipid metabolism, inhibits inflammatory reaction and possesses the potential of enhancing plaque stability.

A Rapid High Throughput Vibration and Vortex-Assisted Matrix Solid Phase Dispersion for Simultaneous Extraction of Four Isoflavones for Quality Evaluation of Semen Sojae Praeparatum.

Isoflavones (daidzein, daidzin, genistein and genistin) were main chemical components and usually selected as markers for quality control of Traditional Chinese Medicine semen sojae praeparatum (SSP). High throughput vibration and vortex-assisted matrix solid phase dispersion and high performance liquid chromatography with diode array detection were developed to simultaneously extract and quantify four isoflavones in SSP. Some parameters influencing extraction efficiency of isoflavones by vortex-assisted matrix solid phase dispersion such as sorbent type, ratio of sample to sorbent, crushing time, vibration frequency, methanol concentration, eluting solvent volume and vortex time were optimized. It was found that the best extraction yields of four isoflavones were obtained when the sample (20 mg) and SBA-3 (40 mg) was crushed by ball mill machine for 2 min at vibration frequency of 800 times per minute. Methanol/water (1.5 ml, 8:2, v/v) solution was dropped into the treated sample and vortexed for 3 min. The recoveries of the four isoflavones ranged from 86.1 to 94.8% and all relative standard deviations were less than 5%. A good linearity (r > 0.9994) was achieved within the range 0.5-125 µg/ml. It was concluded that the high throughput vibration and vortex-assisted matrix solid-phase dispersion coupled with high performance liquid chromatography was user-friendly extraction and quantification method of multiple isoflavones for quality evaluation of SSP.

Simultaneous Determination of Phenolic Acids, Anthraquinones, Flavonoids, and Triterpenes of Cynomorii Herba in Different Harvest Times by LC-MS/MS.

A sensitive and accurate method was developed for the simultaneous determination of twelve components including phenolic acids (gallic acid, protocatechuic aldehyde, protocatechuic acid, and ferulic acid), flavonoids (catechin, epicatechin, rutin, luteolin, luteolin-7-glucoside, and epicatechin gallate), anthraquinones (emodin), and triterpenes (ursolic acid) in Cynomorii herba in different harvest times by liquid chromatography tandem mass spectrometry (LC-MS/MS). The chromatographic separation was achieved on an Eclipse plus C18 (3.0 mm × 50 mm, 1.8 µm) column at 40°C. The mobile phase consisted of acetonitrile and 0.05% formic acid with a gradient elution at a flow rate of 0.4 mL·min -1. Under the optimized conditions, there was good linear relation (r ≥ 0.9944) and satisfactory precision (RSD values less than 5.65%). The recoveries of the twelve components were in the range of 93.5-105%. Moreover, the limits of detection (LOD) ranged from 0.003 to 21 ng mL-1 for the twelve analytes. In conclusion, the validated method was successfully applied to analyze the change regularity of the twelve components of Cynomorii herba in different harvest times. It provides a theoretical basis for choosing the suitable harvesting time of Cynomorii herba.

A Metabolomics Coupled With Chemometrics Strategy to Filter Combinatorial Discriminatory Quality Markers of Crude and Salt-Fired Eucommiae Cortex.

Eucommiae Cortex is commonly used for treating various diseases in a form of the crude and salt-fired products. Generally, it is empirical to distinguish the difference between two types of Eucommiae Cortex. The metabolomics coupled with chemometrics strategy was proposed to filter the combinatorial discriminatory quality markers for precise distinction and further quality control of the crude and salt-fired Eucommiae Cortex. The metabolomics data of multiple batches of Eucommiae Cortex samples was obtained by ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). Orthogonal partial least-squares discriminant analysis was utilized to filter candidate markers for characterizing the obvious difference of the crude and salt-fired Eucommiae Cortex. The accuracy of combinatorial markers was validated by random forest and partial least squares regression. Finally, eleven combinatorial discriminatory quality markers from 67 identified compounds were rapidly screened, identified, and determined for distinguishing the difference between crude and salt-fired Eucommiae Cortex. It was demonstrated that UHPLC-MS based metabolomics with chemometrics was a powerful strategy to screen the combinatorial discriminatory quality markers for distinguishing the crude and salt-fired Eucommiae Cortex and to provide the reference for precise quality control of Eucommiae Cortex.

Recent advances of green pretreatment techniques for quality control of natural products.

A few advancing technologies for natural product analysis have been widely proposed, which focus on decreasing energy consumption and developing an environmentally sustainable manner. These green sample pretreatment and analysis methods following the green Analytical Chemistry (GAC) criteria have the advantage of improving the strategy of chemical analyses, promoting sustainable development to analytical laboratories, and reducing the negative effects of analysis experiments on the environment. A few minimized extraction methodologies have been proposed for replacing the traditional methods in the quality evaluation of natural products, mainly including solid-phase microextraction (SPME) and liquid phase microextraction (LPME). These procedures not only have no need for large numbers of samples and toxic reagent, but also spend a small amount of extraction and analytical time. This overview aims to list out the main green strategies on the application of quality evaluation and control for natural products in the past 3 years.

Network pharmacology strategy for revealing the pharmacological mechanism of pharmacokinetic target components of San-Ye-Tang-Zhi-Qing formula for the treatment of type 2 diabetes mellitus.

ETHNOPHARMACOLOGICAL RELEVANCE: San-Ye-Tang-Zhi-Qing formula (SYTZQ) is an effective prescription for the treatment of pre-diabetes disorders of glycolipid metabolism in type 2 diabetes mellitus (T2DM). It consists of five Chinese herbs including Mori Folium, Nelumbinis Folium, Crataegi Folium, Salviae Miltiorrhizae Radix et Rhizoma and Paeoniae Radix Rubra. AIM OF THE STUDY: This study was aimed to reveal the pharmacological mechanism of pharmacokinetic target components of SYTZQ for the treatment of T2DM. MATERIALS AND METHODS: A rapid, precise and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to quantify simultaneously nuciferin, vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, paeoniflorin and rosmarinic acid in rat plasma after oral administration of SYTZQ. The network pharmacology was used to analyze the effect of the compounds absorbed into the blood of SYTZQ on T2DM. The effects of paeoniflorin, nuciferine and rosmarinic acid on adipogenic differentiation were validated in vitro experiments. RESULTS: The separation was performed on an ACQUITY UHPLC HSS T3 column (2.1 mm × 100 mm, 1.7 µm) using acetonitrile and 0.1% (v/v) formic acid in water as the mobile phase in gradient elution. The calibration curves of five analytes showed good linearity (r ≥ 0.9991) with the lower limits of quantification (LLOQ) between 0.3 and 5.0 ng/mL. The recoveries and matrix effects of five analytes ranged from 81.1% to 113%. The RSDs of inter-day and intra-day precision were all within 13.7%. The validated method was successfully applied to the pharmacokinetic study of five ingredients after oral administration of SYTZQ to rat. 39 major targets and 22 candidate pathways of five compounds absorbed into the blood of rats after administration of SYTZQ were identified and successfully constructed a compound-target-disease-pathway network. It was confirmed that paeniforin, nuciferine and rosmarinic acid could decrease the adipogenicity differentiation in vitro experiments. CONCLUSIONS: The pharmacokinetic parameters indicated that the five components (nuciferin, vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, paeoniflorin and rosmarinic acid) were absorbed and eliminated quickly in vivo. These five absorbed components were associated with 22 pathways, including insulin resistance, regulation of lipolysis in adipocytes, PI3k/AKT-, TNF-, cAMP- and cGMP-PKG-signaling pathway. Paeoniflorin, nuciferine and rosmarinic acid have the effect of inhibiting adipocyte differentiation. This study could provide more reference for quality control, and provide a firm basis for evaluating the clinical efficiency of SYTZQ.

Thrombin-based discovery strategy of bioactive-chemical quality marker combination for pollen of Typha orientalis by metabolomics coupled with chemometrics.

BACKGROUND: It is of utmost significance to choose the bioactive components as quality markers for ensuring the effectiveness of traditional Chinese medicine (TCM). Nonetheless, some markers are able to assess effectively the quality of TCM without considering the pharmacological mechanisms and intrinsic chemical complexities. OBJECTIVE: This underscores the need to discover new and efficient markers which can assess both quality and mechanism of action. Herein, a strategy of bioactive-chemical quality marker combination was proposed to improve the level of the quality control of TCM by metabolomics coupled with chemometrics. METHODS: A four-step plan was followed. Firstly, acquisition of metabolic features and component characterization of different batches of pollen of Typha orientalis C.Presl were performed using UHPLC-Q-TOF/MS. Secondly, the direct inhibitory effects of pollen of T. orientalis on thrombin was assessed by using chromogenic substrate method together with HPLC. Thereafter, bioactive-chemical marker combination associated with anti-thrombin segregation was screened using supervised classifiers. Finally, quantitative assay and prediction-model of selected markers were established for guarantying the quality of pollen of T. orientalis. RESULTS: A total of 22 compounds were annotated based on comparison with previous work from pollen of T. orientalis by UHPLC-Q-TOF/MS. Citric acid and linolenic acid inhibited the thrombin activity with IC50 values, 0.52 ± 0.02 and 0.51 ± 0.02 mg/mL, respectively. A bioactive-chemical marker combination including citric acid, linolenic acid, typhaneoside, and isorhamnetin-3-O-neohesperidoside were discovered and selected as quality markers for evaluation of pollen of T. orientalis according to their capacity for inhibiting thrombin. CONCLUSION: The thrombin-based discovery strategy of bioactive-chemical marker combination was a powerful tool for screening the quality markers for evaluation of pollen of T. orientalis.

An environmentally friendly sodium dodecyl sulfate-synergistic microwave-assisted extraction followed by ultra-high-performance liquid chromatography with photodiode array method for the simultaneous extraction and determination of iridoids, phenylpropanoids, and lignans from Eucommiae Cortex.

A simple and green sodium dodecyl sulfate-synergistic microwave-assisted extraction method was developed to extract and determine the iridoids, phenylpropanoids, and lignans in Eucommiae Cortex followed by ultra-high-performance liquid chromatography with photodiode array detection. The biodegradable solution (sodium dodecyl sulfate) was used as a promising alternative to organic solvents. The response surface methodology provided the optimum extraction conditions (2 mg/mL sodium dodecyl sulfate, 1100 W microwave power, and 6 min extraction time). The recoveries of three types of components ranged from 95.0 to 105% (RSDs < 5%). The intra- and inter-day precision and accuracy were less than 3.40% and within the range of 97.1-105%, respectively. Compared with other extraction methods, this newly established method was more efficient and environmental friendly. The results demonstrated that sodium dodecyl sulfate-synergistic microwave-assisted extraction followed by ultra-high-performance liquid chromatography with photodiode array method was applicable for the simultaneous extraction and determination of these three types of compounds for quality evaluation of Eucommiae Cortex.

Comparative Pharmacokinetics of Geniposidic Acid, Genipin-1-ß-Gentiobioside, Geniposide, Genipin, and Crocetin in Rats after Oral Administration of Crude Gardeniae Fructus and Its Three Processed Products Using LC-MS/MS.

The extract of Gardeniae Fructus (GF) with different processing methods processed the different medicinal properties and efficacy. Crude GF (CGF) could be processed into stir-frying GF (SGF), gancao mix-frying GF (GCGF), and ginger mix-frying GF (GIGF) in practice. An LC-MS/MS method was established for simultaneous quantification of geniposidic acid, geniposide, genipin-1-ß-gentiobioside, genipin, and crocetin in the rat plasma. The LLOQs for determination of all five components were 10 ng/mL. The accuracies of intraday and interday were in the range of 91%-105%. The recoveries of 5 analytes ranged from 81.0% to 114% with RSD less than 14%. The results showed that the AUCs (area under the plasma concentration-time curve) and C max (maximum plasma concentration) of geniposidic acid, genipin-1-ß-gentiobioside, and geniposide after oral administration of the CGF extract were apparently higher than those after oral administration of other processed extracts. C max of geniposide in plasma after administration of GIGF significantly decreased (p < 0.01). Genipin was not detected in rat plasma after administration of the GIGF extract, but it can be detected in plasma after administration of CGF, SGF, and GCGF extract. Furthermore, crocin I and crocin II were not detected in plasma samples. Crocetin had higher concentration in rat plasma versus lower contents in extract. It was demonstrated that the different processing methods might influence the pharmacokinetics of geniposidic acid, genipin-1-ß-gentiobioside, geniposide, genipin, and crocetin.