Multi-omics and chemical profiling approaches to understand the material foundation and pharmacological mechanism of sophorae tonkinensis radix et rhizome-induced liver injury in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Sophorae tonkinensis Radix et Rhizoma (STR) is an extensively applied traditional Chinese medicine (TCM) in southwest China. However, its clinical application is relatively limited due to its hepatotoxicity effects. AIM OF THE STUDY: To understand the material foundation and liver injury mechanism of STR. MATERIALS AND METHODS: Chemical compositions in STR and its prototypes in mice were profiled by ultra-performance liquid chromatography coupled quadrupole-time of flight mass spectrometry (UPLC-Q/TOF MS). STR-induced liver injury (SILI) was comprehensively evaluated by STR-treated mice mode. The histopathologic and biochemical analyses were performed to evaluate liver injury levels. Subsequently, network pharmacology and multi-omics were used to analyze the potential mechanism of SILI in vivo. And the target genes were further verified by Western blot. RESULTS: A total of 152 compounds were identified or tentatively characterized in STR, including 29 alkaloids, 21 organic acids, 75 flavonoids, 1 quinone, and 26 other types. Among them, 19 components were presented in STR-medicated serum. The histopathologic and biochemical analysis revealed that hepatic injury occurred after 4 weeks of intragastric administration of STR. Network pharmacology analysis revealed that IL6, TNF, STAT3, etc. were the main core targets, and the bile secretion might play a key role in SILI. The metabolic pathways such as taurine and hypotaurine metabolism, purine metabolism, and vitamin B6 metabolism were identified in the STR exposed groups. Among them, taurine, hypotaurine, hypoxanthine, pyridoxal, and 4-pyridoxate were selected based on their high impact value and potential biological function in the process of liver injury post STR treatment. CONCLUSIONS: The mechanism and material foundation of SILI were revealed and profiled by a multi-omics strategy combined with network pharmacology and chemical profiling. Meanwhile, new insights were taken into understand the pathological mechanism of SILI.

Mass spectrometry-based ginsenoside profiling: Recent applications, limitations, and perspectives.

Ginseng, the roots of Panax species, is an important medicinal herb used as a tonic. As ginsenosides are key bioactive components of ginseng, holistic chemical profiling of them has provided many insights into understanding ginseng. Mass spectrometry has been a major methodology for profiling, which has been applied to realize numerous goals in ginseng research, such as the discrimination of different species, geographical origins, and ages, and the monitoring of processing and biotransformation. This review summarizes the various applications of ginsenoside profiling in ginseng research over the last three decades that have contributed to expanding our understanding of ginseng. However, we also note that most of the studies overlooked a crucial factor that influences the levels of ginsenosides: genetic variation. To highlight the effects of genetic variation on the chemical contents, we present our results of untargeted and targeted ginsenoside profiling of different genotypes cultivated under identical conditions, in addition to data regarding genome-level genetic diversity. Additionally, we analyze the other limitations of previous studies, such as imperfect variable control, deficient metadata, and lack of additional effort to validate causation. We conclude that the values of ginsenoside profiling studies can be enhanced by overcoming such limitations, as well as by integrating with other -omics techniques.

Determination of anthocyanins, organic acids, and phenolic acids in hibiscus market products using LC/UV/MS.

Hibiscus sabdariffa has gained increasing attention from consumers as a natural, healthy food ingredient, leading to a myriad of available products, yet there is a lack of understanding of the quality and chemical diversity among commercially available hibiscus products. Here, we conducted a survey on the chemistry of 29 hibiscus products (calyces, beverages, and extracts). UHPLC-DAD and UHPLC-QQQ/MS methods with high sensitivity and selectivity were developed to evaluate the chemical profiles pertaining to the sensory attributes (color and taste). Two major anthocyanins (delphinidin-3-sambubioside and cyanindin-3-sambubioside), eight organic acids, and 23 phenolic acids were identified and quantified in hibiscus market products. The results showed that hibiscus samples contained < 0.001-2.372% of total anthocyanins, 0.073-78.002% of total organic acids, and 0.001-1.041% of total phenolic acids, and demonstrated significant variations in market products. This is the first time that an in-depth organic acid profiling was conducted on hibiscus products using UHPLC-QQQ/MS. This method can also be extended to chemical profiling, sensory analysis, quality control, authentication, and standardization of other natural products.

Neuroprotective potential of Erigeron bonariensis ethanolic extract against ovariectomized/D-galactose-induced memory impairments in female rats in relation to its metabolite fingerprint as revealed using UPLC/MS.

Erigeron bonariensis is widely distributed throughout the world's tropics and subtropics. In folk medicine, E. bonariensis has historically been used to treat head and brain diseases. Alzheimer's disease (AD) is the most widespread form of dementia initiated via disturbances in brain function. Herein, the neuroprotective effect of the chemically characterized E. bonariensis ethanolic extract is reported for the first time in an AD animal model. Chemical profiling was conducted using UPLC-ESI-MS analysis. Female rats underwent ovariectomy (OVX) followed by 42 days of D-galactose (D-Gal) administration (150 mg/kg/day, i.p) to induce AD. The OVX/D-Gal-subjected rats received either donepezil (5 mg/kg/day) or E. bonariensis at 50, 100, and 200 mg/kg/day, given 1 h prior to D-Gal. UPLC-ESI-MS analysis identified 42 chemicals, including flavonoids, phenolic acids, terpenes, and nitrogenous constituents. Several metabolites, such as isoschaftoside, casticin, velutin, pantothenic acid, xanthurenic acid, C18-sphingosine, linoleamide, and erucamide, were reported herein for the first time in Erigeron genus. Treatment with E. bonariensis extract mitigated the cognitive decline in the Morris Water Maze test and the histopathological alterations in cortical and hippocampal tissues of OVX/D-Gal-subjected rats. Moreover, E. bonariensis extract mitigated OVX/D-Gal-induced Aß aggregation, Tau hyperphosphorylation, AChE activity, neuroinflammation (NF-κBp65, TNF-α, IL-1ß), and apoptosis (Cytc, BAX). Additionally, E. bonariensis extract ameliorated AD by increasing α7-nAChRs expression, down-regulating GSK-3ß and FOXO3a expression, and modulating Jak2/STAT3/NF-ĸB p65 and PI3K/AKT signaling cascades. These findings demonstrate the neuroprotective and memory-enhancing effects of E. bonariensis extract in the OVX/D-Gal rat model, highlighting its potential as a promising candidate for AD management.

Data on LC-MS profile of Brucea javanica (L.) Merr. and the NMR data of its major indole alkaloids.

This article presents two types of phytochemical data obtained from Brucea javanica (L.) Merr. roots, a medicinal plant belonging to the Simaroubaceae family. The high-resolution LC-MS dataset comprised the chemical profile of dichloromethane extract, which was utilised to annotate 35 chemical constituents. For annotations, the measured spectral data were compared with the in-silico spectral data generated from 920 molecular structures previously reported in Simaroubaceae. Indole alkaloids, quassinoids, aliphatics and lignan were the chemical groups identified in the root extract. The second dataset provides NMR spectra (1H, 13C, COSY, HMQC and HMBC) for the six indole alkaloids previously detected in LC-MS analysis and isolated through centrifugal partition chromatography. The chemical structures of all compounds were confirmed based on NMR data as bruceolline J (compound 7), canthin-6-one-N-oxide (compound 10), bruceolline E (compound 15), 5-methoxycanthin-6-one (compound 16), canthin-6-one (compound 20), and 1­hydroxy-11-methoxycanthin-6-one (compound 22). This phytochemical data was generated to support an ongoing anti-cancer and anti-dengue study.

Chemical profiling of botanical extracts obtained in NADES systems using centrifugal partition chromatography combined with 13 C NMR dereplication-Hypericum perforatum as a case study.

INTRODUCTION: Natural deep eutectic solvents (NADES) have emerged as interesting extractants to develop botanical ingredients. They are nontoxic and biodegradable, nonflammable, easy to prepare, and able to solubilize a wide range of molecules. However, NADES extracts remain difficult to analyze because the metabolites of interest stay highly diluted in the nonvolatile viscous NADES matrix. OBJECTIVE: This study presents a robust analytical workflow for the chemical profiling of NADES extracts. It is applied to Hypericum perforatum aerial parts extracted with the neutral mixture fructose/glycerol/water (3/1/1, w/w/w), and compared to the chemical profiling of a classical dry methanol extract. METHODS: Exploiting polarity differences between metabolites, the H. perforatum NADES extract was partitioned in a liquid-liquid solvent system to trap the hydrophilic NADES constituents in the lower phase. The upper phase, containing a diversity of secondary metabolites from H. perforatum, was fractionated by centrifugal partition chromatography. All fractions were chemically investigated using a 13 C NMR dereplication method which involves hierarchical clustering analysis of the whole NMR dataset, a natural metabolite database for metabolite identification, and 2D NMR analyses for validation. Liquid chromatography-mass spectrometry (LC-MS) analyses were also performed to complete the identification process. RESULTS: A range of 21 metabolites were unambiguously identified, including glycosylated flavonols, lactones, catechins, phenolic acids, lipids, and simple sugars, and 15 additional minor extract constituents were annotated by LC-MS based on exact mass measurements. CONCLUSION: The proposed identification process is rapid and nondestructive and provides good prospects to deeply characterize botanical extracts obtained in nonvolatile and viscous NADES systems.

Antioxidant and anti-Alzheimer activities of Clivia miniata (Lindl) roots, bulbs, and aerial parts: In-vitro and in-silico studies.

Clivia miniata (Lindl) is a member of the family Amaryllidaceae known for its chemically diverse alkaloids with a wide range of biological activities. Many reports revealed a direct role of oxidative stress in the early stage of Alzheimer's disease (AD). Meanwhile, ß-site amyloid precursor protein cleavage enzyme 1 (BACE-1) is a molecular target for the treatment of AD. We aimed to investigate C. miniata root, bulb, and aerial part chemical profiling, antioxidant, BACE-1, and AChE enzyme inhibitory activities. Results showed that the total root had the most potent radical scavenging activity as compared to the total bulb and aerial part, respectively. Ethanol root extract had the most potent BACE-1 inhibitory activity (IC50 = 0.02 ± 0.001 µg/mL) as compared to the bulb and aerial part (IC50 = 0.93 ± 0.13, 1.80 ± 0.24 µg/mL), respectively. Moreover, the total root extract mitigated AChE enzyme activity more than total bulb and aerial fractions with IC50 values of (0.06 ± 0.02, 0.58 ± 0.3, and 1.89 ± 0.42 µg/mL, respectively. Bioassay-guided acid-base fractionation confirmed superior BACE-1 inhibitory activity of the root fractions particularly, methylene chloride and ethyl acetate fractions with (IC50 values of 0.21 ± 0.60 and 0.01 ± 0.001 µg/mL), respectively. UPLC-MS analysis of ethyl acetate and methylene chloride fractions of C. miniata root led to the identification of eight phenolics and thirteen alkaloids, respectively. Molecular docking studies against BACE-1 protein revealed that lycorine di-hexoside, miniatine, and cliviaaline were the most promising hits. Further investigation of anti-AD potential of the aforementioned small molecules is required.

LC-MS based metabolic profiling and wound healing activity of a chitosan nanoparticle-loaded formula of Teucrium polium in diabetic rats.

Healing of wounds is the most deteriorating diabetic experience. Felty germander (Teucrium polium) possesses antioxidant, anti-inflammatory and antimicrobial activities that could accelerate wound healing. Further, nanohydrogels help quicken healing and are ideal biomaterials for drug delivery. In the current study, the chemical profiling, and standardization of T. polium methanolic extract by LC-ESI/TOF/MS/MS and quantitative HPLC-DAD analyses were achieved. The wound healing enhancement in diabetic rats by T. polium nanopreparation (TP-NP) as chitosan nanogel (CS-NG) and investigating the potential mechanisms were investigated. The prepared hydrogel-based TP-NP were characterized with respect to particle size, zeta potential, pH, viscosity, and release of major components. LC-ESI/TOF/MS/MS metabolomic profiling of T. polium revealed the richness of the plant with phenolic compounds, particularly flavonoids. In addition, several terpenoids were detected. Kaempferol content of T. polium was estimated to be 7.85 ± 0.022 mg/ g of dry extract. The wound healing activity of TP-NP was explored in streptozotocin-induced diabetic rats. Diabetic animals were subjected to surgical wounding (1 cm diameter). Then they were divided in 5 groups (10 each). These included Group 1 (untreated control rats), Group 2 received the vehicle of CS-NG; Group 3 (0.5 g of TP prepared in hydrogel), Group 4 (0.5 g of TP-NP), Group 5 represented a positive control treated with 0.5 g of a commercial product. All treatments were applied topically for 21 days. Application of TP-NP on skin wounds of diabetic animals accelerated the healing process as evidenced by epithelium regeneration, formation of granulation tissue followed by epidermal proliferation, along with keratinization as verified by H&E. This was confirmed through enhanced collagen synthesis, as shown by raised hydroxyproline content and Col1A1 gene expression. Moreover, TP-NP significantly alleviated wound oxidative burst and diminished the expressions of inflammatory biomarkers. Meanwhile, TP-NP could enhance the expressions of transforming growth factor beta1 (TGF-ß1), in addition to the angiogenic markers; vascular endothelia growth factor A (VEGFA) and platelet-derived growth factor receptor alpha (PDGFRα). Collectively, chitosan nanogel of T. polium accelerates wound healing in diabetic rats, which could be explained - at least partly - through alleviating oxidative stress and inflammation coupled with pro-angiogenic capabilities.

Unveiling the bioactive potential of Pimpinella anisum L. leaf extracts: Chromatographic profiling, antimicrobial efficiency, and cytotoxicity analysis.

In various countries, Pimpinella has been used to cure several diseases for centuries. Therefore, we focus on one of its potent species in this research. The aim of this experimental study was to document the various extracts derived from Pimpinella anisum that can effectively eradicate oral pathogens. In addition, the presence of antioxidants, antimicrobials, and cytotoxicity was determined using chromatographic testing methods. The alkaloid range was from 22.34 ± 043 mg/g, and the saponin range was from 15.1 ± 1.07 mg/g. HPLC analysis showed that the samples contained eight identified phenolic compounds. The antibacterial activity of ethanolic extract exhibited the highest inhibition region against Streptococcus iniae (43 ± 0.6 mm) and the lowest inhibition region against Staphylococcus haemolyticus (19 ± 0.2 mm) in 200 mg/mL of leaf ethanolic extracts. The antifungal activity revealed that ethanol showed the maximum inhibition zone against Aspergillus luchuensis (42.5 ± 0.19 mm) and the minimum inhibition zone against Aspergillus kawachii (15 ± 0.13 mm) in 200 mg/mL. The current study suggested that, after the isolation of individual components, P. anisum be investigated for assessing biological activity. The mixture and various combinations of these compounds may indicate a truly potent agent that is novel in its ability to combat a wide range of bacteria and oral pathogens.

Background and Perspectives on the Utilization of Canes' and Bunch Stems' Residues from Wine Industry as Sources of Bioactive Phenolic Compounds.

Viticulture activity produces a significant amount of grapevine woody byproducts, such as bunch stems and canes, which constitute potential sources of a wide range of phenolic compounds (PCs) with purported applications. Recently, the study of these byproducts has been increased as a source of health-promoting phytochemicals. Antioxidant, antimicrobial, antifungal, and antiaging properties have been reported, with most of these effects being linked to the high content of PCs with antioxidant properties. This Review summarizes the data related to the qualitative and quantitative composition of PCs recovered from canes and bunch stems side streams of the wine industry, the influence that the different environmental and storage conditions have on the final concentration of PCs, and the current reported applications in specific technological fields. The objective is to give a complete valuation of the key factors to consider, starting from the field to the final extracts, to attain the most suitable and stable characterized product.

A Perspective on Missing Aspects in Ongoing Purification Research towards Melissa officinalis.

Melissa officinalis L. is a medicinal plant used worldwide for ethno-medical purposes. Today, it is grown everywhere; while it is known to originate from Southern Europe, it is now found around the world, from North America to New Zealand. The biological properties of this medicinal plant are mainly related to its high content of phytochemical (bioactive) compounds, such as flavonoids, polyphenolic compounds, aldehydes, glycosides and terpenes, among many other groups of substances. Among the main biological activities associated with this plant are antimicrobial activity (against fungi and bacteria), and antispasmodic, antioxidant and insomnia properties. Today, this plant is still used by society (as a natural medicine) to alleviate many other illnesses and symptoms. Therefore, in this perspective, we provide an update on the phytochemical profiling analysis of this plant, as well as the relationships of specific biological and pharmacological effects of specific phytochemicals. Currently, among the organic solvents, ethanol reveals the highest effectiveness for the solvent extraction of precious components (mainly rosmarinic acid). Additionally, our attention is devoted to current developments in the extraction and fractionation of the phytochemicals of M. officinalis, highlighting the ongoing progress of the main strategies that the research community has employed. Finally, after analyzing the literature, we suggest potential perspectives in the field of sustainable extraction and purification of the phytochemical present in the plant. For instance, some research gaps concern the application of cavitation-assisted extraction processes, which can effectively enhance mass transfer while reducing the particle size of the extracted material in situ. Meanwhile, membrane-assisted processes could be useful in the fractionation and purification of obtained extracts. On the other hand, further studies should include the application of ionic liquids and deep eutectic solvents (DES), including DESs of natural origin (NADES) and hydrophobic DESs (hDES), as extraction or fractionating solvents, along with new possibilities for effective extraction related to DESs formed in situ, assisted by mechanical mixing (mechanochemistry-based approach).

Antibacterial and anti-Toxoplasma activities of Aspergillus niger endophytic fungus isolated from Ficus retusa: in vitro and in vivo approach.

Emergent records propose that Aspergillus niger endophytic fungus is a vital source for various bioactive molecules possessing many biological properties. The current study was designed to inspect the antibacterial and anti-Toxoplasma potentials of Ficus retusa-derived endophytic fungi. After isolation and identification (using 18S rRNA gene sequencing) of A. niger endophytic fungus, LC/MS was utilized for identification and authentication of the chemical profile of the A. niger endophyte extract. Then, the fungal extract was assessed for its antibacterial and antibiofilm activities against Klebsiella pneumoniae clinical isolates. Additionally, its efficacy against Toxoplasma gondii was elucidated in vivo. The fungal extract displayed antibacterial activity against K. pneumoniae isolates with minimum inhibitory concentration values of 64-512 µg/mL. It also possessed a membrane potential dissipating effect using flow cytometry. Moreover, it formed distorted cells with rough surfaces and deformed shapes using a scanning electron microscope (SEM). Regarding its antibiofilm activity, it resulted in a dysregulation of the genes encoding biofilm formation (fimH, mrkA and mrkD) using qRT-PCR in nine K. pneumoniae isolates. The in vivo anti-Toxoplasma potential was demonstrated by decreasing the mortality rate of mice and reducing the tachyzoites' count in the peritoneal fluids and liver impression smears of mice. In addition, the deformities of the parasite decreased, as revealed by SEM and the inflammation in tissues diminished. Thus, A. niger endophytic fungi could be a valuable source of antibacterial and anti-Toxoplasma compounds.

Comprehensive chemical profiling and quantitative analysis of ethnicYi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS.

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.

An integrated approach of high-performance liquid chromatography-mass spectrometry-based chemical profiling, network pharmacology, and molecular docking to reveal the potential mechanisms of Qishen Gubiao granules for treating coronavirus disease 2019.

Qishen Gubiao granules, a traditional Chinese medicine preparation composed of nine herbs, have been widely used to prevent and treat coronavirus disease 2019 with good clinical efficacy. In the present study, an integrated strategy based on chemical profiling followed by network pharmacology and molecular docking was employed, to explore the active components and potential molecular mechanisms of Qishen Gubiao granules in the therapy of coronavirus disease 2019. Using the ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry technique, a total of 186 ingredients corresponding to eight structure types in Qishen Gubiao preparation were identified or structurally annotated with the elucidation of the fragmentation pathways in the typical compounds. The network pharmacology analysis screened 28 key compounds including quercetin, apigenin, scutellarein, luteolin and naringenin acting on 31 key targets, which possibly modulated signal pathways associated with immune and inflammatory responses in the treatment of coronavirus disease 2019. The molecular docking results observed that the top 5 core compounds had a high affinity for angiotensin-converting enzyme 2 and 3-chymotrypsin-like protease. This study proposed a reliable and feasible approach for elucidating the multi-components, multi-targets, and multi-pathways intervention mechanism of Qishen Gubiao granules against coronavirus disease 2019, providing a scientific basis for its further quality evaluation and clinical application.

Integrated transcriptomics and lipidomics investigation of the mechanism underlying the gastrointestinal mucosa damage of Loropetalum chinense (R.Br.) and its representative component.

BACKGROUND: Loropetalum chinensis (R.Br) Oliv (Bhjm), a Chinese folk herbal medicine, was traditionally used in the treatment of wound bleeding and skin ulcers. A new drug named JIMUSAN granules used for gastrosia was developed by our group, and clinical trials have been approved. However, as the principal herb, the material basis and underlying mechanisms of Bhjm in attenuating gastrointestinal mucosa damage (GMD) remain to be systemically illuminated. PURPOSE: An integrated strategy was used to explore the therapeutic effects and mechanisms of Bhjm and ellagic acid (EA) on GMD zebrafish, using network pharmacology, transcriptomics, lipidomics, and real-time quantitative PCR (RT-qPCR) verification. METHODS: First, network pharmacological analysis was used to infer the major effective constituents and targets of Bhjm. Ultra high performance liquid chromatography-linear ion trap/orbitrap high resolution mass spectrometry (UHPLC-LTQ-Orbitrap HRMS) and ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) were employed to identify the chemical constituents and quantify the different types of constituents. Second, zebrafish model of GMD was established by using 2,4,6-trinitrobenzenesulfonic acid (TNBS) to evaluate the efficacy of Bhjm and EA. The potential mechanism was examined by integrated transcriptomics and lipidomics analysis. Finally, validation tests were implemented using RT-qPCR. RESULTS: In this study, targets indentified by network pharmacology were related to inflammation and mucosal damage. Ten representative components that interacted with these targets were simultaneously determined by UHPLC-MS/MS. Sixty four compounds were identified or tentatively characterized, most of which were flavonoids and polyphenols. Bhjm and EA alleviated mucosal damage and reduced inflammation in a TNBS-induced zebrafish GMD model, indicating that EA was the main active compounds. Eight common differentially expressed genes were downregulated by Bhjm and EA, as determined by transcriptomics analysis. Lipidomics analysis confirmed 12 differential lipids, including phosphatidylcholine (PC) and triglyceride (TG). Further network enrichment analysis demonstrated that differential lipid metabolism was regulated by klf4 and hist1h2ba, and were validated by RT-qPCR. CONCLUSION: In our study, the chemical profile of Bhjm was clarified. Moreover, the GMD repair effect and the mechanism of Bhjm and EA was comprehensively analyzed for the first time, involving inflammation and lipid metabolism. Collectively, these findings will be significantly helpful for deeply exploring the clinical application value of Bhjm.

Identification of compounds that enhance bitterness of coffee brew.

The bitterness perception of coffee is a key attribute that impacts consumer acceptance. Nontargeted liquid chromatography/mass spectrometry (LC/MS) flavoromics analysis was applied to identify compounds that enhance the bitter perception of roasted coffee brew. Orthogonal partial least squares (OPLS) analysis was used to model the comprehensive chemical profiles and sensory bitter intensity ratings of fourteen coffee brews with good fit and predictivity. Five compounds that were highly predictive and positively correlated to bitter intensity were selected from the OPLS model, further isolated, and purified using preparative LC fractionation. Sensory recombination testing demonstrated that five compounds significantly enhanced the bitter perception of coffee when presented as a mixture, but not when presented individually. In addition, a set of roasting experiments revealed the five compounds were generated during the coffee roasting process.

Morphology and mass spectrometry-based chemical profiling of peltate glandular trichomes on Mentha haplocalyx Briq leaves.

Mentha haplocalyx Briq (M. haplocalyx) is a herbaceous plant that has long been used as a food, medicinal spice, and flavoring agent in traditional Chinese medicine. Its secondary metabolites, having high commercial values, are mainly produced in tiny specialized structures called glandular trichomes (GTs). The primary purpose of this study was to examine the morphology and metabolites of peltate GTs in M. haplocalyx.Peltate GTs possessed globular dome shapes and intense auto-fluorescence on the surfaces of M. haplocalyx leaves. Structure subsidence and cuticle rupture were found throughout the aging stage of peltate GTs. According to histochemical staining results, the secretion of peltate GTs contained anthraquinone, flavonoids, phenolic acid and terpenoids. In M. haplocalyx peltate GTs and leaf tissues without peltate glandular trichomes, ten and two volatile compounds were identified respectively. Peltate GTs contained 42 non-volatile chemicals with a variety of structural types, including 20 flavonoids, 17 phenolic acids,1 diterpene, 3 anthraquinone and 1 alkane. Meanwhile, 15 non-volatile compounds were discovered in leaf tissues without peltate glandular trichomes, and they were all included in the list of peltate GTs' 41 components. Therefore, Peltate GTs were shown to be the primary site of not just volatile compounds but also non-volatile chemicals in M. haplocalyx. This study provides an important theoretical basis and technical approach for clarifying the bio-active metabolite biosynthesis in M. haplocalyx.

Chemical profiling and quantification analysis of flavonoids in different varieties of Euryales Semen by ultra-high-performance liquid chromatography with tandem mass spectrometry.

Euryales Semen was a traditional Chinese medicine, which has been commonly used to treat spermatorrhea, enuresis, and frequent urination. Flavonoids were a critical ingredient in determining the function and quality of Euryales Semen. At present, no effective method has been established for the qualitative of Euryales Semen flavonoids. In this study, an ultra-high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry method was established for flavonoids. By comparison with standard or literature data, 32 flavonoid compounds have been identified in Euryales Semen. Based on the qualitative results, an ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectroscopy method was developed for the main components, and the linearity, the limit of detection, limit of quantification, repeatability, precision, stability, and recovery of the method were verified. The principal component analysis and the hierarchical clustering heatmaps analysis showed that the 30 batches of samples were distinctly separated into the North Gordon Euryale and South Gordon Euryale, and the measured contents of the six flavonoids in North Gordon Euryale were more abundant than in South Gordon Euryale, especially isoquercitrin, hesperetin, and quercetin. It provided a scientific basis for the quality control of Euryales Semen and a theoretical basis for the rational utilization of Euryales Semen resources.

Diversity of Citrullus colocynthis (L.) Schrad Seeds Extracts: Detailed Chemical Profiling and Evaluation of Their Medicinal Properties.

Seeds and fruits of Citrullus colocynthis have been reported to possess huge potential for the development of phytopharmaceuticals with a wide range of biological activities. Thus, in the current study, we are reporting the potential antimicrobial and anticancer properties of C. colocynthis seeds extracted with solvents of different polarities, including methanol (M.E.), hexane (H.E.), and chloroform (C.E.). Antimicrobial properties of C. colocynthis seeds extracts were evaluated on Gram-positive and Gram-negative bacteria, whereas, anticancer properties were tested on four different cell lines, including HepG2, DU145, Hela, and A549. All the extracts have demonstrated noteworthy antimicrobial activities with a minimum inhibitory concentration (MIC) ranging from 0.9-62.5 µg/mL against Klebsiella planticola and Staphylococcus aureus; meanwhile, they were found to be moderately active (MIC 62.5-250 µg/mL) against Escherichia coli and Micrococcus luteus strains. Hexane extracts have demonstrated the highest antimicrobial activity against K. planticola with an MIC value of 0.9 µg/mL, equivalent to that of the standard drug ciprofloxacin used as positive control in this study. For anticancer activity, all the extracts of C. colocynthis seeds were found to be active against all the tested cell lines (IC50 48.49-197.96 µg/mL) except for the chloroform extracts, which were found to be inactive against the HepG2 cell line. The hexane extract was found to possess the most prominent anticancer activity when compared to other extracts and has demonstrated the highest anticancer activity against the DU145 cell line with an IC50 value of 48.49 µg/mL. Furthermore, a detailed phytoconstituents analysis of all the extracts of C. colocynthis seeds were performed using GC-MS and GC-FID techniques. Altogether, 43 phytoconstituents were identified from the extracts of C. colocynthis seeds, among which 21, 12, and 16 components were identified from the H.E., C.E., and M.E. extracts, respectively. Monoterpenes (40.4%) and oxygenated monoterpenes (41.1%) were the most dominating chemical class of compounds from the hexane and chloroform extracts, respectively; whereas, in the methanolic extract, oxygenated aliphatic hydrocarbons (77.2%) were found to be the most dominating chemical class of compounds. To the best of our knowledge, all the phytoconstituents identified in this study are being reported for the first time from the C. colocynthis.

Comprehensive chemical profiling and quantitative analysis of ethnicYi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS / 中国天然药物

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.