Exploring anti-inflammatory and antioxidant-related quality markers of Artemisia absinthium L. based on spectrum-effect relationship.

INTRODUCTION: Artemisia absinthium L. is a well-known medicinal, aromatic, and edible plant with important medicinal and economic properties and a long history of use in treating liver inflammation and other diseases; however, there has been insufficient progress in quality control. OBJECTIVE: This study aimed to investigate the quality markers for the anti-inflammatory and antioxidant activities of A. absinthium based on spectrum-effect relationship analysis. MATERIALS AND METHODS: Eighteen batches of A. absinthium from different origins were used. Chemical fingerprints were obtained by ultra-performance liquid chromatography (UPLC). The chemical compositions were identified by quadrupole-Orbitrap high-resolution mass spectrometry. Anti-inflammatory activity was assessed by inhibition of cyclooxygenase-2 and 15-lipoxygenase in vitro and inhibition of nitric oxide release in lipopolysaccharide-induced BV-2 cells. Antioxidant activity was assessed by DPPH and ABTS radical scavenging assays. The relationship between bioactivity and chemical fingerprints was then analyzed using chemometrics including gray relational analysis, bivariate correlation analysis, and orthogonal partial least squares analysis. RESULTS: Different batches of A. absinthium extracts possessed significant anti-inflammatory and antioxidant activities to varying degrees. Eighty compounds were identified from A. absinthium, and 12 main common peaks were obtained from the UPLC fingerprints. P3 (chlorogenic acid), P5 (isochlorogenic acid A), and P6 (isochlorogenic acid C) were screened as the most promising active compounds by correlation analysis and further validated for their remarkable anti-inflammatory effects. CONCLUSION: This is the first study to screen the quality markers of A. absinthium by establishing the spectrum-effect relationship, which can provide a reference for the development of quality standards and further research on A. absinthium.

Systematic qualitative analysis of terpenes in mastic (Pistacia lentiscus L.) extract and their fragmentations by UHPLC-Q-Orbitrap-HRMS.

INTRODUCTION: Mastic is a natural resin produced by Pistacia lentiscus L. (Anacardiaceae). The beneficial properties of this resin are attributed to its triterpenes and volatile compounds. OBJECTIVE: This study was conducted to screen and characterize the terpenes in mastic ethyl acetate extract (M-Ex). METHODS: An ultrahigh-performance liquid chromatography coupled to quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) method was developed for the qualitative analysis of terpenes in M-Ex. We utilized in-house-isolated compounds as reference substance (Rs), including monoterpenes (A) with α-pinane structures, tetracyclic triterpene (B) containing tirucallane skeletons, and pentacyclic triterpene (C) belonging to olean, moronic, amyrone, and lupane types. Based on the mass spectrometric characteristics of the above compounds, and the difference in characteristic diagnostic fragment ions (DFIs) in isomeric compounds, the terpene compounds were further identified in M-Ex. RESULTS: Out of a total of 70 compounds, including monoterpenes and tetra-, and pentacyclic triterpenes, 20 were accurately determined by Rs, retention time (RT), and DFIs. Based on the cleavage patterns summarized from the above 20 compounds and with reference to the reported literature, another 50 compounds were putatively identified. Based on our discovery, six terpenic acids with A-seco-tirucallane types and one monoterpene dimer were identified for the first time in mastic. CONCLUSION: Our research serves not only as a foundation for the rapid identification and screening of terpene compounds in mastic but also as a supplementary basis for the identification of such compounds in other types of resins.

Spectrum-effect relationship between ultra-performance liquid chromatography fingerprints and melanogenic effect of Vernonia anthelmintica effective part.

The fingerprint of Vernonia anthelmintica effective part (VAEP) from 15 different producing areas was established, followed by cluster analysis and principal component analysis. The relationship between the fingerprint and the melanogenesis-promoting activity of VAEP was then analyzed using the grey correlation degree and the orthogonal partial least square method. The characteristic peaks reflecting the pharmacodynamic effect of VAEP were identified as vernodalin, 3,5-O-dicaffeoyl quinic acid (3,5-diCQA), and butin. Based on the distribution characteristics of these components in plants from different habitats and the verification of results from the spectrum-effect relationship, vernodalin and 3,5-diCQA can be used as characteristic components for quality control and pharmacodynamic assessment of V. anthelmintica products. This research establishes a theoretical foundation for planting areas and provides a scientific evaluation of the melanogenesis-promoting effect of V. anthelmintica.

Characterisation and identification of chemical constituents in aqueous extract of Fomes officinalis Ames based on ultrahigh-performance liquid chromatography tandem quadrupole-Orbitrap high-resolution mass spectrometry.

INTRODUCTION: Fungal species are an attractive resource for physiologically functional food and drug precursor. Fomes officinalis Ames, a medicinal fungus, is traditionally used as a folk medicine in traditional Chinese medicine prescription for the therapy of cough and asthma. The water-soluble substances in Chinese herbal medicines are likely to play an important physiological function. However, information on probing and identifying chemical components of the aqueous extract of Fomes officinalis Ames (AFO) remains unknown. OBJECTIVE: This study was conducted to screen and characterise the chemical components of AFO. MATERIAL AND METHODS: An effective and sensitive ultrahigh-performance liquid chromatography tandem quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) method with the Full MS/PIL/dd-MS2 acquisition approach was applied for the profiling of chemical components in AFO. An HSS T3 column was used for component separation, and a strategy of simultaneous targeted and untargeted multicomponent characterisation was implemented. Multiple identification approaches were used, including accurate molecular mass and elemental composition matching, literature and database searching, and fragmentation rules elucidation. RESULTS: A total of 115 components, including 20 amino acids and derivatives, six nucleobases, nine nucleosides, 75 dipeptides, two tripeptides, and three other components, were tentatively identified. Among them, the targeted exploring method screened six nucleobases and nine nucleosides including modified nucleosides. To our best knowledge, this is the first time a report has been done on the presence of the 115 compounds in AFO. CONCLUSION: Profiling and characterisation compounds of AFO enriched its material basis, which would lay the foundation for improving potential medicinal and nutritional values and effecting comprehensive quality control of Fomes officinalis Ames.

Rapid identification of chemical components in Xuelian granule by UHPLC-Q-orbitrap-HRMS based on enzyme activity in vitro.

BACKGROUND: Xuelian granule (XL), a traditional Chinese medicine (TCM) formula, has been used for the treatment of diabetic nephropathy for a long time as a hospital preparation. Because the active ingredients in the XL that can help to treat diabetic nephropathy are still unclear, which limits the interpretation for its pharmacological mechanism, further development and subsequent study on the material basis of its efficacy. METHODS: In this study, a screening method based on inhibition activity against aldose reductase (AR) was employed for activity-directed chemical analysis of XL using ultra-high performance liquid chromatography combined with quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-orbitrap-HRMS) technique. RESULTS: A total of 178 compounds, including 46 terpenes, 47 organic acids, 25 flavonoids, 29 phenylethanoid glycosides, and 31 other types, were tentatively identified from XL which might responsible for its AR inhibition activity. CONCLUSION: This is the first study for a systematic, rapid, and accurate qualitative analysis of XL. This research provides a scientific and experimental basis for further researches on pharmacodynamics material basis and quality control of XL.

Profiling of chemical constituents of Matricarla chamomilla L. by UHPLC-Q-Orbitrap-HRMS and in vivo evaluation its anti-asthmatic activity.

Matricarla chamomilla L. is native to European countries and widely cultivated in China, especially in Xinjiang. It has been used in Uygur medicine for the treatment of cough caused by asthma. In this study, UHPLC-Q-Orbitrap-MS was used to detect and identify the components from the active fraction of M. Chamomile, 64 compounds were identified by combining the standards, related literatures and mass spectrometry fragments, including 10 caffeoyl quinic acids, 38 flavonoids, 8 coumarins, 5 alkaloids and 3 other compounds. Furtherly, the anti-asthma activity of active fraction of M. Chamomile was investigated in OVA-induced allergic asthma rat model. The results showed that the number of EOS in Penh and bronchoalveolar lavage fluid (BALF) in the group of the active fraction of M. Chamomile was significantly lower than that in the model group. Besides, the active fraction of M. Chamomile can significantly reduce the IgE level and increased glutathione peroxidase (GSH-Px) in the serum of OVA-induced rats, and ameliorated OVA-induced lung injury. Hence, M. Chamomile could be used to treat asthma through their in vivo antioxidant and anti-inflammatory effects. This study explored the potential material basis of M. Chamomile for the treatment of asthma.

Pharmacokinetic Study and Metabolite Identification of CAM106 in Rats by Validated UHPLC-MS/MS.

Given the limitations of existing antiviral drugs and vaccines, there is still an urgent need for new anti-influenza drugs. CAM106, a rupestonic acid derivative, was studied for its potent antiviral activity and showed a favorable inhibitory effect on influenza virus replication. However, many gaps exist in preclinical studies of CAM106. This study focused on the pharmacokinetic profile and metabolites of CAM106 in vivo. An efficient and fast bioanalytical method was successfully developed and validated for the quantitation of CAM106 in rat plasma. A mobile phase aqueous solution (A, containing 0.1% formic acid) and acetonitrile (B) worked within 0-3.5 min, with 60% B. The mass spectrum scanning mode was the parallel reaction monitoring (PRM) with a resolution of 17,500. The linear range of the method was 2.13-1063.83 ng/mL. The validated method was applied to a pharmacokinetic study in rats. The matrix effects ranged from 93.99% to 100.08% and the recovery ranged from 86.72% to 92.87%. The intra- and inter-day precisions were less than 10.24% and the relative error (RE) ranged from -8.92% to 7.1%. The oral bioavailability of CAM106 was 1.6%. Thereafter, its metabolites in rats were characterized using high-resolution mass spectrometry. The isomers M7-A, M7-B, M7-C, and M7-D were well separated. As a result, a total of 11 metabolites were identified in the feces, urine, and plasma of rats. The main metabolic pathways of CAM106 were oxidation, reduction, desaturation, and methylation. The assay was reliable and provided useful information for further clinical studies of CAM106.

Spectrum-effect relationship between UPLC fingerprints and melanogenic effect of Ruta graveolens L.

A total of 29 batches of R. graveolens were used in this study, their fingerprints were obtained by ultra-performance liquid chromatography (UPLC) and their melanogenesis activities were evaluated. The common peaks were identified by quadrupole-orbitrap high-resolution mass spectrometry (Q-Orbitrap-HRMS). Eleven coumarins, six alkaloids, three flavonoids, three phenolic acids, and four other compounds were found. The spectrum-effect relationships between R. graveolens' chemical fingerprints, the melanin synthesis, and tyrosine's activation activities were established through chemometrics methods which in detail principal component analysis (PCA), gray correlation analysis (GRA), bivariate correlation analysis (BCA) and orthogonal partial least squares analysis (OPLS). The results showed that P18 (bergapten), P22 (isoimperatorin), P15 (kokusaginine), P7 (rutin), P12 (psoralen), and P13 (graveolinine) were relevant to intracellular melanin synthesis activity and tyrosinase activity. Among them, P18 (bergapten), P15 (kokusaginine), and P12 (psoralen) were validated with good melanogenesis activities. This study provides a research basis for future quality control and medicinal application of R. graveolens.

Comprehensive Identification of Chemical Fingerprint and Screening of Potential Quality Markers of Aloe vera (L.) Burm. f. from Different Geographical Origins via Ultra-High-Performance Liquid Chromatography Hyphenated with Quadrupole-Orbitrap-High-Resolution Mass Spectrometry Combined with Chemometrics.

An integrated strategy was developed for the systematic chemical fingerprint and chemometrics analysis for the quality assessment of Aloe vera (L.) Burm. f. The ultra-performance liquid chromatography fingerprint was established, and all common peaks were tentatively identified by using ultra-high-performance liquid chromatography hyphenated with quadrupole-orbitrap-high-resolution mass spectrometry. Afterwards, the datasets of common peaks were subjected to hierarchical cluster analysis, principal component analysis and partial least squares discriminant analysis to holistically compare the differences. The results revealed that the samples were predicted to fall into four clusters, which were related to four different geographical locations. Using the proposed strategy, aloesin, aloin A, aloin B, aloeresin D and 7-O-methylaloeresin A were rapidly determined to be the potential characteristic quality markers. Finally, five screened compounds in 20 batches of samples were simultaneously quantified, and their total contents were ranked as follows: Sichuan province > Hainan province > Guangdong province > Guangxi province, which suggests that geographical origins may be an important factor affecting the quality of A. vera (L.) Burm. f. This new strategy can not only be used to explore possibly the latent active substance candidates for pharmacodynamic studies, but it is also an efficient analytical strategy for other complex traditional Chinese medicine systems.

Pharmacokinetic Study and Metabolite Identification of 1-(3'-bromophenyl)-heliamine in Rats.

Tetrahydroisoquinolines have been widely investigated for the treatment of arrhythmias. 1-(3'-bromophenyl)-heliamine (BH), an anti-arrhythmias agent, is a synthetic tetrahydroisoquinoline. This study focuses on the pharmacokinetic characterization of BH, as well as the identification of its metabolites, both in vitro and in vivo. A UHPLC-MS/MS method was developed and validated to quantify BH in rat plasma with a linear range of 1-1000 ng/mL. The validated method was applied to a pharmacokinetic study in rats. The maximum concentration Cmax (568.65 ± 122.14 ng/mL) reached 1.00 ± 0.45 h after oral administration. The main metabolic pathways appeared to be phase-I of demethylation, dehydrogenation, and epoxidation, and phase II of glucuronide and sulfate metabolites. Finally, a total of 18 metabolites were characterized, including 10 phase I metabolites and 8 phase II metabolites. Through the above studies, we have gained a better understanding of the absorption and metabolism of BH in vitro and in vivo, which will provide us with guidance for future in-depth studies on this compound.

Mass Defect Filtering-Oriented Identification of Resin Glycosides from Root of Convolvulus scammonia Based on Quadrupole-Orbitrap Mass Spectrometer.

This work aimed to develop and evaluate a post-acquisition data processing strategy, referred to as a mass defect filter (MDF), for rapid target the resin glycosides in root of Convolvulus scammonia by setting mass rang and mass defect range from high-resolution MS data. The full-scan mass data were acquired by high-performance liquid chromatography coupled with Q Exactive Plus hybrid quadrupole-orbitrap mass spectrometer that featured high resolution, mass accuracy, and sensitivity. To screen resin glycosides, three parent filter m/z 871, m/z 853, and m/z 869 combined with diagnostic fragment ions (DFIs) approach were applied to remove the interference from complex herbal extract. The targeted components were characterized based on detailed fragment ions. Using this approach, 80 targeted components, including 22 glycosidic acids and 58 resin glycosides were tentatively identified. The present results suggested that the proposed MDF strategy would be adaptable to the analysis of complex system in relevant filed.

Chemical profiling of composite prescription caraway and quantification of three pairs isomeric components in caraway administered rat plasma by tandem mass spectrometry.

Caraway, a well-known traditional Uyghur medicine, has been used to treat vitiligo for centuries. Its biological effects on melanin synthesis of caraway have been investigated. However, beyond psoralen and isopsoralen alone, no further chemical component of caraway has been revealed. In this study, ultra-high performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry was employed to comprehensively characterize the chemical components present in caraway. Based on accurate mass measurements, key fragmental ions and comparison with reference standards, 75 chemical components were identified in caraway. Moreover, a tandem mass spectrometry method was developed and validated for quantitative analysis of three pairs isomeric components, namely psoralen/isopsoralen, bavachin/isobavachalcone and bavachromene/isobavachromene in rat plasma. Psoralen, isopsoralen, bavachin, and isobavachalcone showed linearity with concentration ranging of 1.0-500.0 ng/ml. The linear ranges for bavachromene and isobavachromene were 0.2-500.0 ng/ml. The accuracies were in ranges of 85%-115% with coefficient of variation errors of less than 15%. Furthermore, the method was applied to quantify the three pairs isomeric components in rats after oral administration of caraway.

Chemical Profiling of Kaliziri Injection and Quantification of Six Caffeoyl Quinic Acids in Beagle Plasma by LC-MS/MS.

Vitiligo is a stubborn multifactorial skin disease with a prevalence of approximately 1% in the global population. Kaliziri, the seeds of Vernonia anthelmintica (L.) Willd., is a well-known traditional Uyghur medicine for the treatment of vitiligo. Kaliziri injections is a Chinese-marketed treatment approved by the China Food and Drug Administration for the treatment of vitiligo. The significant effects of Kaliziri injection have been thoroughly studied. However, chemical components studies and plasma quantification studies are lacking for Kaliziri injection. Ultra-high-performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry was employed to comprehensively characterize the caffeoyl quinic acid derivatives present in Kaliziri injection. Based on accurate mass measurements, key fragmental ions and comparisons with reference standards, 60 caffeoyl quinic acid derivatives were identified in Kaliziri injections, including caffeoyl quinic acids, coumaroyl caffeoyl quinic acids, dicaffeoyl quinic acids, feruloyl caffeoyl quinic acids, and dicaffeoyl quinic acid hexosides. Moreover, an HPLC-MS/MS method was developed and validated for the quantitative analysis of 5-caffeoyl quinic acid, 4-caffeoyl quinic acid, 1,3-dicaffeoyl quinic acid, 3,4-dicaffeoyl quinic acid, 3,5-dicaffeoyl quinic acid and 4,5-dicaffeoyl quinic acid in beagle plasma. The quantitative HPLC-MS/MS method was applied to quantify these six major caffeoyl quinic acids in beagle plasma after the subcutaneous administration of Kaliziri injection. All of the six analytes reached their peak plasma of concentrations within 30 min.

Comprehensive chemical profile and quantitative analysis of the Shabyar tablet, a traditional ethnic medicine prescription, by ultra-high-performance liquid chromatography with quadrupole-orbitrap high-resolution mass spectrometry.

The Shabyar tablet is commonly used as a traditional ethnic medicine prescription for the treatment of night blindness, poor vision, and headaches. However, the chemical components of the Shabyar tablet have not been holistically explored, which seriously hinders the discovery of the activity. This study qualitatively and quantitatively investigated the overall chemical profile of the Shabyar tablet using ultra-high-performance liquid chromatography hyphenated with quadrupole-orbitrap high-resolution mass spectrometry. Altogether, 170 chemical components, including 59 flavonoids, 78 organic acids, 12 anthranones, three anthraquinones, one naphthalene, and 17 other compounds were tentatively identified and attributed, with 40 among these being unambiguously characterized in comparison with their corresponding authentic standards. To further determine the major representative constituents of the Shabyar tablet, a quantitative method was used for the simultaneous analysis of 33 characteristic components in Shabyar samples. The results were validated in terms of linearity, precision, repeatability, stability, and recovery. This newly developed approach could be successfully employed for evaluating the holistic quality of crude extracts and Chinese medicines in the Shabyar compound tablet and provide a solid chemical foundation for additional investigations on in vivo pharmacodynamics and therapeutic mechanisms to identify the potential effective components of traditional medicines.

Phenolic Compounds and Bioactivities from Pomegranate (Punica granatum L.) Peels.

Pomegranate peels, which are normally processed as the main byproduct of pomegranate juice production, are worthy of being researched and utilized for the aim of economic and environmental benefits. In a phytochemical investigation of the peels of Punica granatum L., 10 phenolic compounds containing a common hexahydroxy diphenol moiety were isolated. Three of them were identified for the first time and named as pomegranatins A-C, and from the other seven known ones, two of them were obtained from pomegranate peels for the first time. Their structures were determined via extensive spectroscopic analysis. Besides, for the sake of preliminarily comprehending their biological activities, in vitro antimicrobial, antioxidant, as well as antitumor assays were detected. In the DPPH antioxidant assay, six compounds presented significant free radical scavenging ability. Two compounds exhibited moderate antimicrobial activities against Candida albicans; one compound could inhibit the proliferation of both C. albicans and Escherichia coli within limits. Four compounds possessed weak antitumor activity toward the Hela cell line without taking into account the bioavailability of ellagitannins. Overall, these results provided further information on the structural diversity of bioactive compounds present in pomegranate peels, as well as on their biological activities.

Simultaneous Quantification of Four Compounds in Rat Plasma by HPLC-MS/MS and Its Application to Pharmacokinetic Study after Oral Administration of Pomegranate Flowers.

Pomegranate flowers (PFs) were reported to possess various biological activities such as antidiabetic, anti-inflammatory and hepatoprotective activities, and using to treat diabetes. Although chemical constituents and pharmacological activities of PFs have been studied, unfortunately, there was no report on the pharmacokinetic profile of PFs in vivo. In this study, a selective high-performance liquid chromatography triple quadrupole tandem mass spectrometry (HPLC-QQQ-MS/MS) method was developed and validated for simultaneous quantification of four compounds (corilagin, ellagic acid, gallic acid and brevifolincarboxylic acid) in rat plasma after oral administration of PFs. The good linearity concentration ranges for the four analytes were from 2.5 to 3000 ng/mL with coefficient value R2 > 0.99 in calibration curves. The intra- and inter-day accuracy of the four analytes was in the range of 85.33-102.50%, with relative standard deviation (RSD) of <14.81%. The stability results showed that accuracy of the four analytes was in the range of 81.88-104.74%, with RSD of <14.86%. The validation method was successfully applied to pharmacokinetic profiles of the four analytes in rats after oral administration of PFs extract. This pharmacokinetic study can provide better understanding to clarify in vivo mechanisms of PFs and may facilitate its further development as therapeutic agent.

Spectrum-effect relationship between UPLC fingerprints and antidiabetic and antioxidant activities of Rosa rugosa.

In this study, the antidiabetic and antioxidant properties of the chemical constituents of Rosa rugosa Thunb. (R. rugosa) was evaluated through analysis of spectrum-effect relationship. The ultra-performance liquid chromatography (UPLC) fingerprints of 21 batches of R. rugosa were evaluated by similarity analysis (SA) and hierarchical clustering analysis (HCA). The 28 common components were identified by ultra-high-performance liquid chromatography coupled to quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-orbitrap-HRMS/MS). Meanwhile, the antidiabetic activities and antioxidant activities of 21 batches of R. rugosa were estimated in vitro. Besides, four chemometrics named principal component analysis (PCA), grey correlation analysis (GRA), partial least squares regression (PLSR) and the bivariate correlations analysis (BCA) were applied to construct spectrum-effect relationship between the UPLC fingerprints and biological activities of R. rugosa. The spectrum-effect relationship study revealed that di-O-galloyl-HHDP-glucoside, galloyl-HHDP-glucoside and avicularin were more relevant to antidiabetic activity. Di-O-galloyl-HHDP-glucoside, galloyl-HHDP-glucoside and ellagic acid were the main antioxidant components of R. rugosa. The current bioassay and spectrum-effect relationships are proper for associating sample quality with the active ingredient, and our finding would provide foundation and further understanding of the quality evaluation and quality control of R. rugosa.

Chemical Components and Hepatoprotective Mechanism of Xwak Granule in Mice Treated with Acute Alcohol.

OBJECTIVE: To evaluate the hepatoprotective mechanism of Xwak granule (Xwak) in treatment of mice with alcoholic liver injury via activating ERK/NF-κB and Nrf/HO-1 signaling pathways. METHODS: The chemical composition of Xwak was tested by liquid chromatography coupled with mass spectrometry (LC-MS). Herein, 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical tests were performed in vitro. The hepatoprotective effect of Xwak was assessed at different concentrations (1.5, 3, and 6 g/kg) in a mouse model of alcoholic liver injury. RESULTS: Totally, 48 compounds, including 16 flavonoids, 8 tannins, 9 chlorogenic acids, and 15 other compounds, were identified from Xwak. Xwak showed to have a satisfactory antioxidant activity in vitro. In a group of Xwak-treated mice, the serum levels of alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) were decreased compared with a group of the mouse model of alcoholic liver injury. In addition, the levels of antioxidant enzymes, such as glutathione peroxidase (GSH-PX), total superoxide dismutase (T-SOD), and catalase (CAT), were noticeably increased and the levels of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), transforming growth factor-ß (TGF-ß), and interleukin-6 (IL-6) were markedly reduced in the liver of mice. The state of oxidative stress in the mouse model of alcoholic liver injury was improved after treatment with Xwak. The improvement of inflammation-mediated disruption may conducive to the Xwak activity in the control of liver injury. The signals of p-ERK1/2, p-NF-κB, COX-2, iNOS, CYP2E1, Nrf, and HO-1 were significantly induced in the liver of mice after treatment with Xwak. CONCLUSIONS: The abovementioned findings indicated that the hepatoprotective mechanism of Xwak could be achieved by activating ERK/NF-κB and Nrf/HO-1 signaling pathways to alleviate oxidative stress and inflammatory.

Comprehensive characterisation of phenolics from Nitraria sibirica leaf extracts by UHPLC-quadrupole-orbitrap- MS and evaluation of their anti-hypertensive activity.

ETHNOPHARMACOLOGICAL RELEVANCE: For more than ten scores years, the leaves and fruits of Nitraria sibirica have been used as a natural remedy for indigestion, irregular manes, and hypertension in the Middle East and Central Asia, especially, are recommended for hypertension treatment in the northwest region, China. AIM OF THE STUDY: we aimed to support the traditional usage of N. sibirica leaves as pharmaceuticals or dietary supplements in treatment of hypertension by investigating their chemical constituents and anti-hypertensive activity. METHODS: We identified the chemical composition of N. sibirica leaves ethanolic purified extract (NSL-EPE) using UHPLC-quadrupole-orbitrap-MS, and quantified the main chemical constituents by an analytical method established and validated. We also evaluated anti-hypertensive activity of NSL-EPE using spontaneously hypertensive rats (SHR): blood pressure was measured weekly by non-invasive blood pressure (NIBP) measurements; hemodynamic parameters, biochemical and clinical chemistry variables in plasma, serum and kidney tissue were measured after 10 weeks of treatment with NSL-EPE as well. RESULTS: UHPLC-quadrupole-orbitrap-MS analysis identified 52 compounds, of which 40 compounds were reported for the first time in N. sibirica. 11 phenolic compounds further quantitatively analyzed, among which the most abundant compound was found to be clovin (8.8%). Systolic blood pressure decreased progressively from the second treatment week compared to that in non-treated SHRs. The plasma endothelin, aldosterone, angiotensin II levels were significantly increased, while the level of NOX was significantly decreased; glutathione to oxidized glutathione ratio, superoxide dismutase and total catalase levels in the kidney tissue were markedly accelerated, while malondialdehyde level was significantly reduced in NSL-EPE treated SHRs. Moreover, the serum cholesterol, triglyceride, blood uria nitrogen and creatinine were attenuated in NSL-EPE treated SHRs (P < 0.05), but in sharp contrast to those values in the water-treated SHRs. CONCLUSION: This study screened out leading compounds from N. sibirica and offered a new understanding of the antihypertensive properties of N. sibirica leaves, by which inhibit oxidative stress-induced endothelial dysfunction and improve lipid profiles.

Simultaneous determination of loading capacity and selectivity in preparative off-line two-dimensional separation: An application for purification of corilagin from Pomegranate flower extracts.

This work describes the development of capacity orthogonal chromatography (COC), a new technique for simultaneously determining the loading capacity and orthogonality during the construction of two-dimensional (2D) separations. Three steps were required for the construction of a COC based on the correlation between the selectivity factor (α) and both orthogonality and loading capacity. (1) α values of the impurities-target compound were used to normalize the retention of the impurities around the target compound. (2) α values were input into four quadrants of a coordinate system to identify correlations between orthogonality and loading capacity. α values of the impurities must be greater in the first dimension than the second dimension, with iterated analyses performed until an αmax is obtained for the two purification methods. (3) Touch-peak separation using the first-dimensional αmax was performed and the target compound was collected. The co-eluted impurities are further separated in the second dimension. To test the efficiency of this technique, a COC using two methods on a standard C18 column was developed to purify corilagin from pomegranate flower extract. Despite its low abundance, 288 mg of corilagin was obtained by COC and further purified by LH-20 gel chromatography to obtain the compound with an 80.0% recovery and 98.4% purity. Compared to COC, the purity of corilagin independently obtained using the same purification methods and identical loading capacity was poor (60.1% and 61.6%). These results indicate that COC is a useful tool for extending loading capacity in the development of preparative 2D separations.