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.

Tunable internal structure carbon sphere synthesis driven by water-solubility and its application in gas separation.

A method for synthesizing carbon spheres with a tunable particle size and internal structure from polyfurfuryl alcohol (PFA) was developed. By tuning the concentration of a structure directing agent (polypropylene glycol, PPG), we found a mechanism to tune the inner architecture of carbon spheres driven by water-solubility. A mixture of PFA and PPG transferred from the "water-in-oil" phase to an "oil-in-water" phase with an increasing content of PPG because of a difference in water-solubility between furfuryl alcohol (FA), PFA, and PPG. As a result, the internal morphology of the carbon sphere evolved from a "cheese-like" to a "pomegranate-like" structure, which was accompanied by an increasing specific surface area and pore volume. Furthermore, the separation of C2H2 and C2H3Cl was tested on the 25%-FACS (furfuryl alcohol-based carbon sphere) sample under different activation treatments with CO2 or CO2-NH3, with the coexisting "cheese-like" and "pomegranate-like" inner structures, owing to its moderate pore volume and mechanical strength. The maximum adsorption capacity of C2H3Cl reached 0.77 mmol g-1, while C2H2 was adsorbed in significantly lower quantities. It is believed that the high polarizability and high dipole moment of the C2H3Cl molecule primarily contribute to the excellent performance of C2H2 and C2H3Cl separation, and the introduction of polar N-containing groups on the carbon skeleton further promotes C2H3Cl adsorption.

Rapid classification and identification of chemical components in three different Zanthoxylum species by ultra-high-performance-liquid chromatography quadrupole-orbitrap-mass spectrometry.

Zanthoxylum, as a medicinal and edible herbal medicine, has a long history and complex chemical composition. There are many varieties of Zanthoxylum, and there are differences in composition between varieties. In this study, a rapid classification and identification method for the main components of Zanthoxylum was established using ultra-high-performance-liquid chromatography quadrupole-orbitrap-mass spectrometry. The components of Shandong Zanthoxylum bungeanum, Wudu Zanthoxylum bungeanum, and Zanthoxylum schinifolium were identified by studying the characteristic fragmentations and neutral losses of characteristic components. A total of 48 common components and 24 different components were identified and the fragmentation patterns of the main components, such as flavonoids, alkaloids, and organic acids were summarized. These findings provided a reference for the study of pharmacodynamic substance basis and quality control of different varieties of Zanthoxylum.

Systematic characterization of the chemical constituents in vitro and in vivo of Qianghuo by UPLC-Q-TOF-MS/MS.

The Chinese herb Qianghuo is an antiphlogistic herb with many effects and complex components. In this study, the chemical composition of Qianghuo and its components in rat plasma after oral administration were investigated using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). The extracts, blank plasma, and plasma containing the drug were analyzed by mass spectrometry, and data collected in both positive and negative ion modes were analyzed using Masslynx software, and the structures were confirmed by combining the compound fragment ions and mass spectrometry cleavage pathways. A total of 62 in vitro chemical components were identified, including 27 coumarins, 18 organic acids, 5 amino acids, 5 glycosides, 2 flavonoids, 4 nucleotides, and 1 ester, which were summarized from the obtained compounds in terms of their possible cleavage patterns. Among the identified 31 compounds in rat plasma, 21 were prototypes, mostly coumarins, organic acids, and flavonoids, and 10 were metabolites, which were mainly generated via hydroxylation and methylation pathways. Based on these, this study provides a theoretical foundation for quality control and basic research on Qianghuo medicinal substances.

Phenotypic flexibility in metabolic adjustments and digestive function in white-shouldered starlings: responses to short-term temperature acclimation.

Changing the intrinsic rate of metabolic heat production is the main adaptive strategy for small birds to cope with different ambient temperatures. In this study, we tested the hypothesis that the small passerine the white-shouldered starling (Sturnus sinensis) can modulate basal metabolism under temperature acclimation by changing the morphological, physiological and biochemical state of its tissues and organs. We measured the effects of temperature on body mass, basal metabolic rate (BMR), wet mass of various internal organs, state 4 respiration (S4R) and cytochrome c oxidase (CCO) activity in the pectoral muscle and organs, metabolites in the pectoral muscle, energy intake, histological dynamics and the activity of duodenal digestive enzymes. Warm acclimation decreased BMR to a greater extent than cold acclimation. At the organ level, birds in the cold-acclimated group had significantly heavier intestines but significantly lighter pectoral muscles. At the cellular level, birds in the cold-acclimated group showed significantly higher S4R in the liver and heart and CCO activity in the liver and kidney at both the mass-specific and whole-organ levels. A metabolomic analysis of the pectoral tissue revealed significantly higher lipid decomposition, amino acid degradation, ATP hydrolysis, and GTP and biotin synthesis in cold-acclimated birds. Acclimation to cold significantly increased the gross energy intake (GEI), feces energy (FE) and digestive energy intake (DEI) but significantly decreased the digestive efficiency of these birds. Furthermore, cold-acclimated birds had a higher maltase activity and longer villi in the duodenum. Taken together, these data show that white-shouldered starlings exhibit high phenotypic flexibility in metabolic adjustments and digestive function under temperature acclimation, consistent with the notion that small birds cope with the energy challenges presented by a cold environment by modulating tissue function in a way that would affect BMR.

A Study on the Regularity of Acupoint Match Based on Association Rules with SP6 as the Main Acupoint and Its Clinical Application.

Objective: This study aims to explore formula patterns and application rules for SP6 as the main acupoint in prescriptions, utilizing association rules. Methods: We conducted an extensive search in databases including China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform (CDDB), China Science and Technology Journal Database (CSTJ), PubMed and Web of Science databases for literature published between January 2013 and June 2023, focusing on acupuncture prescriptions with SP6 as the main acupoint for various diseases. Inclusion and exclusion criteria were applied for literature screening. Relevant data was extracted, creating a database. Acupoints in conjunction with SP6 were analyzed using SPSS Modeler 18.0 and Cytoscape 3.7.2 software for acupoints appearing ≥15 times. Gephi software constructed a complex network model. The frequency of acupuncture points was analyzed to summarize the composition rules and clinical application rules of acupuncture points. Results: A total of 902 articles met inclusion criteria, yielding 672 prescriptions with SP6 as the main acupoint, paired with 197 different acupoints including ST36, CV4, and LI4. Neurological, obstetric, and gynecologic, as well as urological diseases, were predominantly treated. Among them, the predominant diseases include insomnia, primary dysmenorrhea, sequelae of stroke, and others, totaling 42 types. Conclusion: SP6-based prescriptions exhibit diverse applications, effectively treating insomnia, post-stroke sequelae, and primary dysmenorrhea. Commonly paired acupoints belong to Conception Vessel, Stomach meridian of foot-yangming, and Governor Vessel, and there are certain rules in their composition.

Rapid classification and identification of chemical constituents in Leonurus japonicus Houtt based on UPLC-Q-Orbitrap-MS combined with data post-processing techniques.

Leonurus japonicus Houtt (LJH) is a bulk medicinal material commonly used in clinical practice, but its complex constituents have not been completely understood, posing challenges to pharmacology, pharmacokinetic research, and scientific and rational drug use. As a result, it is critical to develop an efficient and accurate method for classifying and identifying the chemical composition of LJH. In this study, ultra-performance liquid chromatography-quadrupole electrostatic field-orbital trap high resolution mass spectrometry (UPLC-Q-Orbitrap-MS) was successfully established, along with two data post-processing techniques, characteristic fragmentations (CFs) and neutral losses (NLs), to quickly classify and identify the chemical constituents in LJH. As a result, 44 constituents of LJH were identified, including four alkaloids, 20 flavonoids, two phenylpropanoids, 17 organic acids, and one amino acid. The method in this paper enables classification and identification of chemical compositions rapidly, providing a scientific foundation for further research on the effective and toxic substances of LJH.

Analysis of chemical components of Jianwei Xiaoshi Tablets based on UPLC-Q-Orbitrap-MS technology.

As a compound preparation of traditional Chinese medicine, Jianwei Xiaoshi Tablets (JXT) is made from five Chinese herbs: Taizishen (Pseudostellariae Radix), Chenpi (Citri Reticulatae Pericarpium), Shanyao (Dioscoreae Rhizoma), Maiya (Hordei Fructus Germinatus) and Shanzha (Crataegi Fructus). It is mainly used to treat dyspepsia. However, the chemical composition of JXT is complex and unclear. In this study, ultra performance liquid chromatography-quadrupole-orbitrap-mass spectrometry and data post-processing technologies were used to analyse the samples of JXT. Firstly, the mass spectrometric information of the main components of five traditional Chinese herbs in JXT was summarised and a compound database was established. Then, the mass spectrometric data detected by the prepared samples was compared with the database. Finally, 93 chemical components were successfully identified, including 6 amino acids, 34 flavonoids, 18 alkaloids, 15 organic acids, 9 cyclic peptides and 11 other components, and the rapid classification and identification of chemical components of JXT were realised.

Comparative studies of immobilized polysaccharide derivatives chiral stationary phases for enantioseparation of furanocoumarins and dihydroflavones and discussion on chiral recognition mechanism.

Enantiomeric separation of furanocoumarins and dihydroflavones compounds were systematically studied in the normal-phase mode using four different polysaccharide-type chiral stationary phases, namely, Chiralpak IA, Chiralpak IC, Chiralpak IG, and Chiralpak IK-3 by high-performance liquid chromatography. The effect of alcohol modifiers and alcohol content on enantiomeric separation was evaluated for the separation of furanocoumarins and dihydroflavones. All the eight compounds have achieved baseline separation with the resolutions ranging between 1.52 and 23.11. For a better insight into the enantiorecognition mechanisms, thermodynamic analysis was carried out. The mechanisms of chiral recognition have been discussed. Among four chiral columns, Chiralpak IG exhibited the most universal and the best enantioseparation ability toward furanocoumarins and dihydroflavones when used n-hexane-isopropanol and n-hexane-ethanol as mobile phase, respectively. The steric hindrance, hydrogen bonding, and π-π interaction played major roles in chiral recognition on Chiralpak IG. By comparing four chiral columns, this work systematically analyzed the separation methods of furanocoumarins and dihydroflavones for the first time and reported some active chiral ingredients of traditional Chinese medicine that have never been separated, which provided a further insight into the enantioseparation of furanocoumarins and dihydroflavones on chiral stationary phases.

Rapid identification of chemical constituents in Hugan tablets by ultra-performance liquid chromatography-quadrupole-exactive orbitrap mass spectrometry.

Hugan tablet is a Chinese medicine preparation. It is composed of Bupleuri Radix, Artemisiae Scopariae Herba, Isatidis Radix, Schisandrae Chinensis Fructus, Suis Fellis Pulvis, and Vigna radiata L. It has the effects of dispersing stagnated liver qi, strengthening the spleen and eliminating food to be used for the treatment of chronic hepatitis and early cirrhosis. However, the chemical composition of Hugan tablet is complex and not fully understood, which hampers the research in pharmacology. In this study, a reliable method for the rapid analysis and identification of the chemical components in Hugan tablet by their characteristic fragments and neutral losses using ultra-performance liquid chromatography-quadrupole-exactive orbitrap mass spectrometry was developed. A total of 144 chemical components were tentatively identified, including 57 organic acids, 19 flavonoids, 23 alkaloids, 18 lignans, 7 saponins, and 20 others. These components may be the active ingredients of Hugan tablet. The established method can systematically and rapidly analyze the chemical components in Hugan tablet, which provides a basis for the pharmacodynamic substance study and is meaningful for the quality control of Hugan tablet.

A practical method for rapid discrimination of constituents in Psoraleae Fructus by UPLC-Q-Orbitrap MS.

Psoraleae Fructus (PF) is one of the most frequently used traditional Chinese medicine, which has good efficacy in warming kidney to activate yang, promoting inspiration to relieve asthma and warming spleen to stop diarrhea. However, the chemical composition of PF is complex, which makes it difficult to determine its active and toxic components. In order to rapidly classify and identify the chemical components of the extracts from PF, this research was processed with CNKI, PubMed, and PubChem databases and data post-processing technique basing on ultrahigh performance liquid chromatography quadrupole orbitrap mass spectrometry (UPLC-Q-Orbitrap MS) technique. Finally, 73 chemical components were discriminated, including 44 flavonoids, 18 coumarins, and 11 terpenoids, with the cleavage rules of each chemical component summarized. This study established a UPLC-Q-Orbitrap MS method for the separation and discrimination of the chemical constituents of PF, which can lay a foundation for the further study of its medicinal substances and quality control.

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.

In-capillary screening and quantifying the antioxidants of Yangxinshi Tablet using field-enhanced sample injection-micellar electrokinetic chromatography.

An in-capillary 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxyradicals combining field-enhanced sample injection with micellar electrokinetic chromatography was developed for screening and determination of the major antioxidants in Yangxinshi Tablet. To obtain simultaneous separation and detection of radicals and analytes, relevant factors were optimized separately. Under the optimum conditions, four compounds including salvianolic acid B, hyperoside, puerarin, and caffeic acid were identified as the major antioxidants. All validation results covering recovery, precision, and stability demonstrated good applicability of the method. On this basis, the total antioxidant activity was successfully evaluated in terms of the decreased peak area of radicals. There was a correlation coefficient of 0.8974 between the total contents of major antioxidants and the total antioxidative activity of the sample. Therefore, these four compounds were selected as combinatorial markers for the quality evaluation of Yangxinshi Tablet. It was concluded that the established method presented a powerful potential to screen and quantify active ingredients in the complex preparation of Chinese medicine.

An integrated strategy combining metabolomics and machine learning for the evaluation of bioactive markers that differentiate various bile.

Animal bile is an important component of natural medicine and is widely used in clinical treatment. However, it is easy to cause mixed applications during processing, resulting in uneven quality, which seriously affects and harms the interests and health of consumers. Bile acids are the major bioactive constituents of bile and contain a variety of isomeric constituents. Although the components are structurally similar, they exhibit different pharmacological activities. Identifying the characteristics of each animal bile is particularly important for processing and reuse. It is necessary to establish an accurate analysis method to distinguish different types of animal bile. We evaluated the biological activity of key feature markers from various animal bile samples. In this study, a strategy combining metabolomics and machine learning was used to compare the bile of three different animals, and four key markers were screened. Quantitative analysis of the key markers showed that the levels of Glycochenodeoxycholic acid (GCDCA) and Taurodeoxycholic acid (TDCA) were highest in pig bile; Glycocholic acid (GCA) and Cholic acid (CA) were the most abundant in bovine and sheep bile, respectively. In addition, four key feature markers significantly inhibited the production of NO in LPS-stimulated RAW264.7 macrophage cells. These findings will contribute to the targeted development of bile in various animals and provide a basis for its rational application.

Enantioseparation and determination of oxypeucedanin and its application to a stereoselective analysis in Angelica Dahuricae Radix and pharmacokinetic study of rats.

In this study, a new enantioseparation method was established for the quantitative analysis of the oxypeucedanin enantiomers by using cellulose tris(3,5-dichlorophenyl carbamate) stationary phase column Chiralpak IC. For this method, enantiomeric separation of oxypeucedanin was achieved with the mobile phase consisting of acetonitrile-water (60:40, v/v) at a flow rate of 0.5 mL/min by changing the type and proportion of mobile phase. And the quantitative determination of racemic oxypeucedanin in Angelica Dahuricae Radix (in vitro) and rat plasma (in vivo) were performed on above-mentioned condition by High PerformanceLiquid Chromatography combined with diode arrangement detector (HPLC-DAD) and mass spectrometry (HPLC-MS/MS). The precision, repeatability, stability, recovery were within the acceptance criteria. And the method was validated in the concentration range of 1-400 µg/mL for the two enantiomers in vitro and 0.2-600 ng/mL in vivo. After validation, the established method was successfully applied to the stereoselective analysis of racemic oxypeucedanin in Angelica dahurica from different regions and the stereoselective pharmacokinetic investigation in rat. Results showed that the (+)-oxypeucedanin was at a relative high level in Angelica dahuricae Radix and (-)-oxypeucedanin performed a higher plasma concentration, which demonstrated the difference of oxypeucedanin enantiomers both in vitro and in vivo.

Rapid classification and identification of chemical components of Astragali radix by UPLC-Q-TOF-MS.

INTRODUCTION: Pharmacological studies indicate that Astragalus (AR) has various bioactivities, including anticancer, antiaging, anti-inflammatory, antiviral, and antioxidant activities. Flavonoids, saponins, amino acids, and polysaccharides are the main active components in AR. However, its complex chemical compositions bring certain difficulties to the analysis of this traditional Chinese medicine (TCM). Therefore, there is an urgent need to establish a method for rapid classification and identification of the chemical constituents in AR. OBJECTIVE: To establish a method for rapid classification and identification of the main components of flavonoids, saponins, and amino acids in AR. METHODS: The samples were analysed with ultra-high-performance liquid chromatography time-of-flight quadrupole mass spectrometry (UPLC-Q-TOF-MS) and data post-processing techniques. Firstly, fragmentation information was obtained in the positive and negative ion modes. Then, to realize the rapid classification and identification of AR components, the characteristic fragmentations (CFs) and neutral losses (NLs) were compared with information described in the literature. RESULTS: A total of 45 chemical constituents were successfully screened out, including 22 flavonoids, 13 saponins, and 10 amino acids. CONCLUSION: The established method realised the efficient classification and identification of flavonoids, saponins, and amino acid compounds in AR, which provided a basis for further study on AR.

A novel reverse migration micellar electrokinetic chromatography method for in-capillary screening and quantifying of antioxidant components in Sanyetangzhiqing using 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxidation-free radical.

A novel method of reverse migration micellar electrokinetic chromatography (RM-MEKC) using 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxidation-free radical was developed to screen the major antioxidants from Sanyetangzhiqing (SYTZQ), which is a new patent drug for diabetes. For simultaneous detection and separation of 2,2'-bis (3-ethylbenzothiazoline-6-sulfonic acid) ammonium salt (ABTS+ ) and chemical components of SYTZQ, the key factors were optimized and the method was validated under the optimal conditions. All the relative standard deviations of recovery, precision, and stability were below 6.6%. Based on these, the RM-MEKC method has been successfully used to screen the main antioxidants from SYTZQ tablets. Five compounds, including rosmarinic acid, salvianolic acid B, lithospermic acid, hyperoside, and isoquercetin, were separated and identified as the major antioxidants. There was a linear relationship with correlation coefficient of 0.923 between the total amount of major antioxidants and total antioxidative activity of SYTZQ. Consequently, these five ingredients could be selected as combinatorial markers for quality control of SYTZQ, and the established method was concluded to be a simple, reliable, and powerful tool to screen and quantify active compounds for quality evaluation of SYTZQ.

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.