Discovery of antitussive material basis and mechanisms in Citri Sarcodactylis Fructus by coupling UHPLC-Q/Orbitrap HRMS combined spectrum-effect relationship and metabolomics analyses.

Citri Sarcodactylis Fructus (CSF) is widely used as food raw material and traditional Chinese medicine. Fingerprints of different fractions of CSF were established for spectrum-effect relationship analysis, and the main compounds were identified by UHPLC Quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q/Orbitrap HRMS). The antitussive effect was evaluated using a classical mouse model of cough induced by ammonia water. One-way ANOVA was used to determine differences in efficacy. The potential active compounds were screened by spectrum-effect relationship with grey relational degree analysis (GRA), Pearson bivariate correlation analysis (Pearson's), and partial least squares analysis (PLS) analyses. Differential metabolites associated with cough in serum were screened and identified using orthogonal partial least squares-discriminant analysis, HMDB database, and UHPLC-Q/Orbitrap HRMS. Metabolic pathway analysis was performed using MetaboAnalyst 5.0. Results indicate that 70 % ethanol elution fraction (70 % EF) is the major active fraction, and 8 components were identified to possess antitussive effects. Metabolomic analysis showed that 19 metabolites are potential biomarkers related to cough, and 70 % EF can remarkable restore 13 of them to normal levels (P < 0.05). These biomarkers are mainly involved in glycerophospholipid metabolism and sphingolipid metabolism. This study aims to reveal the main pharmacodynamic active sites and potential active ingredients of CSF's antitussive effect. In addition, metabolomics was used to preliminarily elucidate the in-vivo regulatory mechanism of the antitussive effect of the 70 % EF of CSF.

Harnessing the Carrier Solvent Complexity of Crop Biostimulant Liquid Formulations Using Locally Available Transesterified Waste Cooking Oil: Economic Recycling, Solvent Performance, and Bioefficacy Evaluation.

Locally sourced waste cooking oil (WCO) was successfully base-catalyzed and transesterified with methanol into biodiesel to produce biostimulant (nitrobenzene) formulations and replace high-risk carrier solvents. Ideal synthesis conditions were composed of 1% NaOH, MeOH/oil molar ratio (6:1), reaction temperature (65 °C), a 3 h mixing rate, and 97-98% yields. Gas chromatography-mass spectrometry (GC-MS) analysis identified five fatty acid methyl esters (FAMEs) including palmitic, linoleic, oleic, stearic, and eicosenoic acids with high solubilization and olfactory characteristics. Using anionic and nonionic emulsifiers in conjunction with recycled biodiesel, a stable emulsifiable concentrate (NB 35% EC) was created with greater storage stability, wettability, and spreading capabilities than those of organic solvent-based ones. The highest counts of fruits per plant (35.80), flowers per plant (60.00), yield per plant (3.56 kg), and yield per hectare (143.7 quintals) were recorded in treatments with 4 mL/L biodiesel-based EC in field bioassays. In addition to having superior biosafety, FAME-based EC exhibits minimal phytotoxicity and is less harmful to aquatic creatures. It was discovered that the average cost-effectiveness was 5.49 times less expensive than solvent-based EC. In order to utilize waste oils as a locally obtained, sustainable alternative solvent with a wide solubilization range, low ecotax profile, circular economy, and high renewable carbon index, this integrative technique was expanded.

Mechanism of Essential Oil of Mongolian Medicine Zhenbaowan in the Treatment of Rheumatoid Arthritis Based on GC-MS and Bioinformatics Technology / 医药导报

Objective To explore the antioxidant capacity of the volatile oil components of the Mongolian medicine Zhenbaowan and the mechanism of action in the treatment of rheumatoid arthritis(RA)by using bioinformatics technology,GC-MS technology,and antioxidant activity experiment.Methods The volatile oil in Zhenbaowan was extracted using water vapor distillation and the volatile oil components were qualitatively analyzed by GC-MS.Based on the results of GC-MS analysis,bioinfor-matics analysis was used to investigate the main components,key targets,and pathways of the volatile oil of Zhenbaowan in pre-venting and treating RA.Meanwhile,the antioxidant activity of the volatile oil was determined and analyzed using the 2,2-Diphen-yl-1-picrylhydrazyl(DPPH)method.Results A total of 115 volatile oil components were identified in Zhenbaowan,of which the top 48 components accounted for 97.66%of the total volatile oil content.Bioinformatics analysis revealed that Zhenbaowan can regulate signal transduction,inflammation,and protein phosphorylation through 147 RA-related targets,and intervene in signaling pathways such as PI3K-Akt,MAPK,and RAS.Molecular docking results showed that the seven main components of Zhenbaowan volatile oil can spontaneously bind to the five core targets.Antioxidant activity tests have proved that the volatile oil of Zhenbaowan has a more significant antioxidant capacity.Conclusion Using GC-MS high-throughput analysis technology combined with bioinformatic analysis and antioxidant activity test,the pharmacodynamic substances and mechanism of action of the volatile oil components of Zhenbaowan,a Mongolian medicine,in the treatment of RA are hypothesized.It provides a theoretical basis for the further study of the volatile oil components of Zhenbaowan.

Species Differentiation and Quality Evaluation for Atractylodes Medicinal Plants by GC/MS Coupled with Chemometric Analysis.

The utilization of rhizomes from the genus Atractylodes has been challenging due to their closely related origins. In this study, we developed an analytical strategy to differentiate Atractylodes lancea (A. lancea), Atractylodes chinensis (A. chinensis), Atractylodes japonica (A. japonica), and Atractylodes macrocephala (A. macrocephala), and compared their volatile compositions. Gas chromatography-mass spectrometry (GC/MS) was used to analyze the volatile profiles of essential oils extracted from 59 batches of samples. Chemometric methods enabled a better understanding of the differences in volatile oils between the four species and identified significant components affecting their classification and quality. A total of 50 volatile components were identified from the essential oils by GC/MS. Unsupervised and supervised chemometric analyses accurately distinguished A. lancea, A. chinensis, A. japonica, and A. macrocephala. Furthermore, five characteristic chemical markers, namely hinesol, ß-eudesmol, atractylon, atractylodin and atractylenolide I, were obtained, and their respective percentage contents in individual species and samples were determined. This study provides a valuable reference for the quality evaluation of medicinal plants with essential oils and holds significance for species differentiation and the rational clinical application of Atractylodes herbs.

Concentrations of polycyclic aromatic hydrocarbons in Vietnamese takeaway coffee: effects of coffee variety, roasting temperature and time.

The research goal was to estimate the level of risk to human health posed by polycyclic aromatic hydrocarbons (PAHs) in Vietnamese takeaway coffee. A variety of roasted coffee beans were collected and tested for the presence of PAHs in various takeaway locations throughout Vietnam. Furthermore, the effect of roasting conditions on PAH concentrations in Vietnamese Robusta coffee was also studied and demonstrated. Gas chromatography-mass spectrometry, a modern, accurate, and fast method, was used to determine the research results. Six PAHs, namely naphthalene, anthracene, pyrene, fluorene, phenanthrene, and benz[a]anthracene, were found in the 100 collected samples, with average concentrations (µg/kg dry weight) of 943.7 ± 40.3, 195.1 ± 4.9, 36.1 ± 1.1, 33.3 ± 2.2, 28.2 ± 1.7, and 2.0 ± 0.1, respectively. It was found that the tested samples were almost free of PAH4 contamination. The research showed that the total value of PAH quantifications in Robusta coffee increased with increasing roasting temperature and decreased with increasing roasting time. In addition, the calculated value of the total hazard quotient (THQ) was less than 1, and the obtained value of the incremental lifetime carcinogenic risk (ILCR) did not exceed 1·10-5, meaning that coffee consumers in Vietnam are safe from exposure to PAHs present in the investigated coffee beans.

Genomics for the characterization of the mechanisms of microbial strains in degrading petroleum pollutants.

Four petroleum-tolerant bacteria, namely, Pseudomonas hibiscicola, Enterobacter hormaechei, Rhizobium pusense and Pseudomonas japonica were isolated. These strains showed excellent performance in the remediation of petroleum contamination with degradation percentages of 26.13%, 26.47%, 32.27%, and 18.74% for petroleum hydrocarbons, 29.63%, 70.11%, 88.38%, and 67.03% for n-docosane, and 61.00%, 96.36%, 98.00%, and 67.01% for fluorene. Accordingly, the strain of Rhizobium pusense was used to further study its underlying degradation mechanism. N-docosane could be metabolised through the pathway of subterminal oxidation by Rhizobium pusense, while the main pathway for fluorene metabolism is the meta-cleavage. R. pusense contains 10 genes that are involved in the synthetic of biosurfactants and 71 genes that are involved in the metabolism of petroleum hydrocarbons and organic pollutants, such as hydrocarbons, toluene, xylene, ethylbenzene and naphthalene. This study was the first to determine that concerning the metabolism ability and metabolic genes of R. pusense for petroleum pollutant degradation, which is important for understanding the bioremediation mechanism of petroleum pollution.

Characterization of active peptides derived from three leeches and comparison of their anti-thrombotic mechanisms using the tail vein thrombosis model in mice and metabonomics.

Background and aims: The increasing incidence of cardiovascular diseases has created an urgent need for safe and effective anti-thrombotic agents. Leech, as a traditional Chinese medicine, has the effect of promoting blood circulation and removing blood stasis, but its real material basis and mechanism of action for the treatment of diseases such as blood stasis and thrombosis have not been reported. Methods: In this study, Whitmania Pigra Whitman (WPW), Hirudo nipponica Whitman (HNW) and Whitmania acranutata Whitman (WAW) were hydrolyzed by biomimetic enzymatic hydrolysis to obtain the active peptides of WPW (APP), the active peptides of HNW (APH) and the active peptides of WAW (APA), respectively. Then their structures were characterized by sykam amino acid analyzer, fourier transform infrared spectrometer (FT-IR), circular dichroism (CD) spectrometer and LC-MS. Next, the anti-thrombotic activities of APP, APH and APA were determined by carrageenan-induced tail vein thrombosis model in mice, and the anti-thrombotic mechanisms of high-dose APP group (HAPP), high-dose APH group (HAPH) and high-dose APA group (HAPA) were explored based on UHPLC-Q-Exactive Orbitrap mass spectrometry. Results: The results showed that the amino acid composition of APP, APH and APA was consistent, and the proportion of each amino acid was few different. The results of FT-IR and CD showed that there were no significant differences in the proportion of secondary structures (such as ß-sheet and random coil) and infrared absorption peaks between APP, APH and APA. Mass spectrometry data showed that there were 43 common peptides in APP, APH and APA, indicating that the three have common material basis. APP, APH and APA could significantly inhibit platelet aggregation, reduce black-tail length, whole blood viscosity (WBV), plasma viscosity (PV), and Fibrinogen (FIB), and prolong coagulation time, including activated partial thrombin time (APTT), prothrombin time (PT) and thrombin time (TT). In addition, 24 metabolites were identified as potential biomarkers associated with thrombosis development. Among these, 19, 23, and 20 metabolites were significantly normalized after administration of HAPP, HAPH, and HAPA in the mice, respectively. Furthermore, the intervention mechanism of HAPP, HAPH and HAPA on tail vein thrombosis mainly involved in linoleic acid metabolism, primary bile acid biosynthesis and ether lipid metabolism. Conclusion: Our findings suggest that APP, APH and APA can exert their anti-blood stasis and anti-thrombotic activities by interfering with disordered metabolic pathways in vivo, and there is no significant difference in their efficacies.

Exploring the Potential of Sago Caterpillars as Cooking Oils: Extraction, Purifying and Characterization.

<b>Background and Objective:</b> The dual demand for palm oil, both as a cooking oil and as a raw material for biodiesel, gives rise to concerns regarding potential shortages. Hence, it is crucial to explore alternative sources of cooking oil, with one such alternative being the oil extracted from sago caterpillars. This study aims to extract and determine the characteristics of sago caterpillar oil and its potential as cooking oil. <b>Materials and Methods:</b> Sago caterpillar oil extraction was done using pressing, Soxhlet extraction and Folch's lipid extraction. The yield of sago caterpillar oil obtained by pressing, Soxhlet and Folch's lipid extraction were 20, 16 and 2.2%, respectively. Oil purifying was done using degumming, neutralization and bleaching. Furthermore, the resulting sago caterpillar oil was characterized physically, chemically and organoleptic. <b>Results:</b> The sago caterpillar oil met the requirements as cooking oil based on the Indonesian National Standard for cooking oil and other chemical parameters. The results of the analysis of sago caterpillar oil with gas chromatography-mass spectrometer showed that the sago caterpillar oil contained 0.15% lauric acid, 2.06% myristic acid, 5.92% palmitoleic acid, 55.05% palmitic acid, 0.84% linoleic acid, 34.00% oleic acid and 1.43% stearic acid. The main peak positions from the fourier transform infrared spectrophotometer are at 725, 1118, 1165, 1234, 1373, 1458, 1743, 2854 and 2924 cm¹. The results of the analysis of sago caterpillar oil showed that the lipid profile of sago caterpillar oil was similar to commercial palm oil. <b>Conclusion:</b> Based on the results of extraction, purifying and characterization, it was concluded that sago caterpillar oil has the potential to be used as cooking oil.

Microscale supercritical fluid extraction combined with supercritical fluid chromatography and proton-transfer-reaction ionization time-of-flight mass spectrometry for a magnitude lower limit of quantitation of lipophilic compounds.

The application of proton transfer ionization reaction mass spectrometry (PTR MS) combined with microscale supercritical fluid extraction (SFE) and supercritical fluid chromatography (SFC) aiming to quantitate single-cell fatty acid analysis levels was investigated. Using a microscale extraction vessel, the obtained low limits of quantitation (LLOQs) of arachidonic acid and arachidic acid were 1.2 and 2.7 fmol, respectively, by using less than 1 µL of sample on stainless steel frit. A series of phthalate, vitamin K1, and α-tocopherol were also tested, and the LLOQ was less than one femtomole for phthalate and 35 and 13 fmol for vitamin K1 and α-tocopherol, respectively. A microliter portion of SFE extracts was introduced into the SFC column by split injection, improving the reproducibility of the chromatography and separation efficiency. The method in the present study has great potential to quantitate lipophilic molecules on the nanogram scale of a sample without complex preparation procedures.

Development and evaluation of a HILIC-MS method for the determination of amino acid and non-amino acid impurities in histidine.

Developing analytical methods to assure and control the quality of amino acids has long been a challenge for food ingredient, dietary supplement, and pharmaceutical industries due to the high polarity and the absence of chromophores in many amino acids; the situation worsens further by the lack of information of impurities that could potentially be introduced during the manufacturing processes. Herein we utilize a four-step strategy including impurity identification, method development, sample analysis, and targeted impurity detection and quantitation to demystify the impurity profiles of amino acids. The effectiveness of the approach is highlighted using histidine as an example. Analysis of histidine manufacturing and degradation processes led to the identification of 12 potential impurities of histidine, including amino acids (arginine, lysine, asparagine, aspartic acid, alanine, and glycine) and non-amino acid impurities (histamine, histidinol, 4-imidazoleacrylic acid, 4-imidazoleacetic acid, ß-imidazolelactic acid, and urea). A HILIC method using Poroshell 120 HILIC-Z column (2.1 × 100 mm, 2.7 µm) and a mobile phase system consisting of ammonium formate buffer at pH 3.2 in water and 0.1% formic acid in acetonitrile coupled with a single quadrupole mass spectrometer was developed for the detection and quantitation of the proposed impurities. Evaluation of 11 commercial histidine samples using the developed method revealed distinct impurity profiles, as a fingerprint for each sample; seven of the 12 proposed impurities were detected in histidine samples tested. The developed method was evaluated in terms of specificity, linearity, range, accuracy, precision, and sensitivity (LOQ: 2.5-60.6 ng/mL) for its suitability for compendial applications. Given the high degree of overlap between the proposed and the detected impurities, the approach could be utilized to strengthen USP standards for controlling the quality of histidine. Extension of the strategy to the analysis of other amino acids is currently underway.

Echinodorus macrophyllus fraction with a high level of flavonoid inhibits peripheral and central mechanisms of nociception.

BACKGROUND AND AIM: Echinodorus macrophyllus (Kunth.) Micheli is popularly used for acute and chronic inflammatory conditions. The anti-inflammatory activity was previously demonstrated for its flavonoid-enriched fractions. The aim of this work assessed the antinociceptive properties of both aqueous extract and its fractions. EXPERIMENTAL PROCEDURE: The antinociceptive activity was determined by acetic acid-induced writhing, formalin test, tail immersion test, hot-plate test, xylene-induced ear edema methods, and the evaluation of its mechanism was performed in the writhing model. The aqueous extract of Echinodorus macrophyllus (AEEm) was fractionated, yielding Fr20, and Fr40. Fr40 composition was determined by HPLC-DAD-ESI-MS. RESULTS AND CONCLUSION: Fr20 (all doses) and Fr40 (100 mg/kg) reduced the nociception in the tail-flick model. Both fractions increased the percentage of maximum possible effect with 25 mg/kg, in the hot-plate assay, at 60 min, while AEEm reduced pain only with 50 and 100 mg/kg. There was a reduction in xylene-edema index, with Fr40 (25 mg/kg), AEEm (50 mg/kg) and Fr20 (50 mg/kg). All doses of AEEm, Fr20, and Fr40 reduced both phases of the formalin model. In the abdominal contortion model, Fr40 presented the highest activity, reducing 96% of contortions and its antinociceptive mechanism was evaluated. The results indicated the involvement of NO and adrenergic activation pathways. The main components of Fr40 are swertisin, swertiajaponin, isoorientin 7,3'-dimethyl ether, swertisin-O-rhamnoside, isoorientin, isovitexin, isovitexin-Orhamnoside, and isovitexin-7-O-glucoside. The aqueous extract of E. macrophyllus leaves and its fractions exhibited significant analgesic effect, mediated through both peripheral and central mechanisms being considered a potentially antinociceptive drug.

DHA turnover in pregnant women using the natural abundance variation of 13C: a pilot study.

The importance of DHA intake to support fetal development and maternal health is well established. In this pilot study we applied the natural abundance approach to determine the contribution of 200 mg/day of DHA supplement to the plasma DHA pool in 19 healthy pregnant women on a free diet.Women received DHA, from pregnancy week 20 until delivery, from an algal source (N=13, Algae group) or from fish oil (N=6, Fish group) with slightly different content of 13C.We measured plasma phospholipids DHA 13C:12C ratio (reported as δ13C) prior to supplementation (T0), after 10 (T1) and 90 days (T2) and prior to delivery (T3).The δ13C of DHA in algae and fish supplements were -15.8±0.2 mUr and -25.3±0.2 mUr (p<0.001).DHA δ13C in the Algae group increased from -27.7±1.6 mUr (T0) to -21.9±2.2 mUr (T3) (p<0.001), whereas there were not significant changes in the Fish group (-27.8±0.9 mUr at T0 and -27.3±1.1 mUr at T3, p=0.09).In the Algae group 200 mg/day of DHA contributed to the plasma phospholipid pool by a median value of 53% (31-75% minimum and maximum). This estimation was not possible in the fish group.Our results demonstrate the feasibility of assessing the contribution of DHA from an algal source to the plasma DHA pool in pregnant women by the natural abundance approach. Plasma δ13C DHA did not change when consuming DHA of fish origin, with almost the same δ13C value of that of the pre-supplementation plasma δ13C DHA.

UPLC-MS/MS method for the determination of the herb composition of Tangshen formula and the in vivo pharmacokinetics of its metabolites in rat plasma.

INTRODUCTION: Tangshen formula (TSF) is a traditional Chinese medicine composed of seven medicinal herbs including Astragalus membranaceus, Rehmannia glutinosa Libosch, Citrus aurantium L., etc. which is used to treat diabetic nephropathy III, IV qi and yin deficiency and stasis syndrome. Most of the studies on TSF are pharmacological and pharmacodynamic experiments. There are few basic studies on its chemical substances, and the effective constituents are not clear. OBJECTIVE: To analyse the main chemical components of TSF and the absorbed components in rat plasma following oral administration based on liquid chromatography tandem mass spectrometry (LC-MS/MS). Moreover, providing a rapid and valid analytical strategy for simultaneous determination of six components in rat plasma and use it in pharmacokinetic studies. RESULTS: A total of 132 components were identified in TSF, and 44 components were identified in rat plasma after oral TSF, 35 of which were prototype components and nine were metabolic components. A sensitive and reliable LC-MS/MS method was developed for simultaneous determination of six components in rat plasma. The intra-day and inter-day precision relative standard deviation (RSD) was lower than 15%; the accuracy of low, medium and high concentrations ranged from 80% to 120%. The recovery met the requirements and the RSD of the recoveries was less than 15%. CONCLUSION: A total of 132 components were identified in TSF. The LC-MS/MS quantitative method for the simultaneous determination of morroniside, loganin, notoginsenoside R1 , ginsenoside Re, ginsenoside Rb1 and astragaloside IV in rat plasma was established for the first time. The pharmacokinetic parameters are clarified, which can guide the clinical medication of TSF.

Comprehensive Identification of Astilbin Metabolites in Rats Based on Multiple Metabolite Templates Combined with UHPLC-Q-Exactive Mass Spectrometry.

BACKGROUND: Astilbin, a dihydroflavonoid compound widely found in plants, exhibits a variety of pharmacological activities and biological effects. However, little is known about the metabolism of this active compound in vivo, which is very helpful for elucidating the pharmacodynamic material basis and application of astilbin. OBJECTIVE: To establish a rapid profiling and identification method for metabolites in rat urine, faeces and plasma using a UHPLC-Q-Exactive mass spectrometer in negative ion mode. METHODS: In this study, a simple and rapid systematic strategy and 7 metabolite templates, which were established based on previous reports, were utilized to screen and identify astilbin metabolites. RESULTS: As a result, a total of 71 metabolites were detected and characterized, among which 32 metabolites were found in rat urine, while 27 and 38 metabolites were characterized from rat plasma and faeces, respectively. These metabolites were presumed to be generated through ring cleavage, sulfation, dehydrogenation, methylation, hydroxylation, glucuronidation, dehydroxylation and their composite reactions. CONCLUSION: This study illustrated the capacity of the sensitive UHPLC-Q-Exactive mass spectrometer analytical system combined with the data-mining methods to rapidly elucidate the unknown metabolism. Moreover, the comprehensive metabolism study of astilbin provided an overall metabolic profile, which will be of great help in predicting the in vivo pharmacokinetic profiles and understanding the action mechanism of this active ingredient.

How to identify "Material basis-Quality markers" more accurately in Chinese herbal medicines from modern chromatography-mass spectrometry data-sets: Opportunities and challenges of chemometric tools.

Modern chromatography - mass spectrometer (MS) technology is an essential weapon in the exploration of traditional Chinese medicines (TCMs) which is based on the "effectiveness-material basis-quality markers (Q-markers)". Nevertheless, the hardware bottleneck and irregular operation will limit the accuracy and comprehensiveness of test results. Chemometrics was thereby used to solve the existing problems: 1) The method of 'design-modeling-optimization' can be adopted to solve the multi-factor and multi-level problems in sample preparation/ parameter setting; 2) The approaches of signal processing can be used to calibrate the deviation from retention time (rt) dimension and mass-to-charge ratio (m/z) dimension in different types of instruments; 3) The methods of multivariate calibration and multivariate resolution can be utilized to analyze the co-eluting peaks in complex samples. When the researchers need to capture essential information on raw data sets extracting the higher level of information on essential features, 1) The significant components which affects the drug properties/efficacy can be find by the pattern recognition and variable selection; 2) Fingerprint-efficacy modeling is explored to clarify the material basis, or to screen out the Q-markers of biological significance; 3) Chemometric tools can apply to integrate chemical (metabolic) fingerprints with network pharmacology, bioinformatics, omics and others from a multi-level perspective. Under these programs, the qualitative/quantitative works will achieve in chemical (metabolic) fingerprint and metabolic trajectories, which leads to an accurate reflection of "material basis and Q-markers" in TCMs. Likewise, an in-depth hidden information can be disclosed, so that the components of drug properties/efficacy will be found. More importantly, multidimensional data can be integrated with fingerprints to acquire more hidden information.

The Study on the Active Ingredients and Potential Targets of Rice Bran Petroleum Ether Extracts for Treating Diabetes Based on Network Pharmacology.

AIM AND OBJECTIVE: In ancient China, rice bran was used to treat diabetes and hyperlipidemia. The aim of this paper is to explore the active compounds and underlying mechanism of Rice Bran Petroleum Ether extracts (RBPE) against diabetes using network pharmacology. MATERIALS AND METHODS: Gas chromatography-mass spectrometer analysis was performed to identify the chemical composition in RBPE. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, Swiss Target Prediction database, BATMAN-TCM, comprehensive database of human genes and gene phenotypes, therapeutic target database, DurgBank and GeneCards database were used to screen targets. The "component-target-disease" interactive network was constructed by Cytoscape software. Gene ontology and pathways related to the targets were analyzed by ClueGO, and core targets were screened by the MCODE, and Autodock vina was used for molecular docking. RESULTS: The compounds with a percentage greater than 1.0% were selected for subsequent analysis. The RBPE contains oleic acid, (E)-9-Octadecenoic acid ethyl ester, and other chemical components that can regulate insulin, mitogen-activated protein kinase 3, epidermal growth factor receptor, mitogen-activated protein kinase 1, and other genes, which were mainly related to Pathways in cancer, Human cytomegalovirus infection and AGE-RAGE signaling pathway in diabetic complications, etc. The affinity of the core compounds and the corresponding protein of the gene targets was good. CONCLUSION: The results of network pharmacology analysis indicate that the RBPE has multiple anti- diabetic ingredients, and RBPE exert anti-diabetic activity through multiple targets and signaling pathways. The present study can provide a scientific basis for further elucidating the mechanism of RBPE against diabetes.

Comparative analysis of chemical constituents and bioactivities of the extracts from leaves, seed coats and embryoids of Ginkgo biloba L.

A total of 25 compounds including terpenoids, flavonoids, biflavonoids and ginkgolic acids were identified and quantified with a reliable, simple, and simultaneous method from Ginkgo leaves, seed coats and embryoids with different tree ages (approximately identified as 25, 500, 1000 and 2000 years). Leaves had the highest amount of total bioactive compounds. Seed coats had moderate contents of flavonoids, which was 15 times higher than embryoids. Furthermore, the effects of tree ages on bioactive compounds differ in three parts. The contents of bilobalide, ginkgolide J, ginkgolide C, ginkgolide B, ginkgolide A in embryoids and seed coats were highest from 500-year-old tree, while in leaves were highest from 25-year-old tree. This work first investigated the extensive bioactive compounds in ginkgo leaves, seed coats and embryoids from Ginkgo trees older than 500-year, it gives good reference for making better use of Ginkgo products.

How to identify “Material basis–Quality markers” more accurately in Chinese herbal medicines from modern chromatography-mass spectrometry data-sets: Opportunities and challenges of chemometric tools / 中草药·英文版

Modern chromatography - mass spectrometer (MS) technology is an essential weapon in the exploration of traditional Chinese medicines (TCMs) which is based on the “effectiveness-material basis-quality markers (Q-markers)”. Nevertheless, the hardware bottleneck and irregular operation will limit the accuracy and comprehensiveness of test results. Chemometrics was thereby used to solve the existing problems: 1) The method of ‘design-modeling-optimization’ can be adopted to solve the multi-factor and multi-level problems in sample preparation/ parameter setting; 2) The approaches of signal processing can be used to calibrate the deviation from retention time (rt) dimension and mass-to-charge ratio (m/z) dimension in different types of instruments; 3) The methods of multivariate calibration and multivariate resolution can be utilized to analyze the co-eluting peaks in complex samples. When the researchers need to capture essential information on raw data sets extracting the higher level of information on essential features, 1) The significant components which affects the drug properties/efficacy can be find by the pattern recognition and variable selection; 2) Fingerprint-efficacy modeling is explored to clarify the material basis, or to screen out the Q-markers of biological significance; 3) Chemometric tools can apply to integrate chemical (metabolic) fingerprints with network pharmacology, bioinformatics, omics and others from a multi-level perspective. Under these programs, the qualitative/quantitative works will achieve in chemical (metabolic) fingerprint and metabolic trajectories, which leads to an accurate reflection of “material basis and Q-markers” in TCMs. Likewise, an in-depth hidden information can be disclosed, so that the components of drug properties/efficacy will be found. More importantly, multidimensional data can be integrated with fingerprints to acquire more hidden information.

[Investigation of chemical profile of bile acids and determination of them in cultured Bovis Calculus with UPLC-QDA].

In this study, 10 batches of samples of cultured Bovis Calculus(cow-bezoar) were determined for the investigation of chemical profile of bile acids with the UPLC-QDA method established. The results showed that nine common bile acids, cholic acid(CA), deoxycholic acid(DCA), chenodeoxycholic acid(CDCA), taurocholic acid(TCA), taurodeoxycholic acid(TDCA), taurochenodeoxycholic acid(TCDCA), glycocholic acid(GCA), glycodeoxycholic acid(GDCA) and glycochenodeoxycholic acid(GCDCA), were founded in samples and 7 bile acids were quantified except GDCA and GCDCA. In these samples, unconjugated bile acids, the major type of bile acids, accounted for more than 97% of all types of bile acids. As for unconjugated bile acids, CA was the most major bile acid in cultured cow-bezoar and it was about twice as much as DCA. The relative low-cost method established in the current study is accurate, rapid and sensitive, which is suitable for the studies of other drugs from animal bile.

Systematic analysis of the metabolites of Roukou Wuwei pills by ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry after oral administration to rats.

Roukou Wuwei pill is one of the most commonly used Mongolian medicinal prescriptions and historically used for the treatment of depression. This research aimed to illustrate the metabolic characteristic of Roukou Wuwei pills in vivo. To address this objective, an ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry method based metabonomics approach was used to detect and analyze the metabolites of Roukou Wuwei pills. The chromatographic separation was completed on an Agilent SB-C18 column (1.8 µm, 2.1 × 50 mm) with a gradient elution system (acetonitrile and 0.1% formic acid-water). Electrospray ionization was operated in a full-scan mode at m/z 100-1000. The data were collected in positive and negative ion modes. The Masslynx 4.1 and SIMCA-P 13.0 software were used to analyze the mass spectrometry data and select the potential metabolites by using an orthogonal partial least-squares discriminant analysis, which was applied to investigate the differences between the blank and drug groups in biosamples of rats. Finally, totally 87 metabolites were detected based on their tandem mass spectrometry data. Among them, 69 metabolites are potential new compounds.