Metabolomic study of the estrogenic and anti-osteoporotic potential of Erythrina bidwillii leaf.

Erythrina bidwillii Lindl., Leguminosae, constitutes a valuable crop for horticulture and medicine; however, it is rarely investigated. Menopause is a crucial transitional period in women's health. Women worldwide consider the use of phytoestrogens as a safe hormone replacement therapy to alleviate detrimental menopausal symptoms. Thus, the discovery of novel phytoestrogens is highly demanded. The present study aimed to investigate, for the first time, the metabolomic profile and the estrogenic potential of E. bidwillii Lindl. leaf. Ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry and gas chromatography-mass spectrometry metabolite profiling revealed the prevalence of alkaloids, flavonoids, isoflavonoids and fatty acids. Additionally, five erythrinan alkaloids, cristanine A (1), 8-oxoerythraline (2), (+)-erythrinine (3), (+)-erythraline (4) and 8-oxoerythrinine (5), along with the isoflavonoid genistin (6), were isolated. Erythrina bidwillii leaf extract exhibited significant in vivo estrogenic, anti-osteoporotic, anti-hyperlipidemic, hepatoprotective, and nephroprotective activities, utilizing ovariectomized rat model. Moreover, ethyl acetate and hexane fractions possessed significant in vitro estrogeic potential on MCF-7 cell lines. An in silico study of the isolated metabolites revealed that (+)-erythrinine (3) and 8-oxoerythrinine (5) exhibited the highest affinity for ERα and ERß, respectively, modeling them as potential estrogenic lead metabolites. Therefore, E. bidwillii leaf could be employed as promising hormone replacement therapy for postmenopausal women after thorough clinical trials.

Comparative metabolic profiling of olive leaf extracts from twelve different cultivars collected in both fruiting and flowering seasons.

Olea europaea is an economically significant crop native to Mediterranean countries. Its leaves exhibit several biological properties associated to their chemical composition. The aqueous ethanolic extracts of olive leaves from twelve different cultivars were analyzed by high performance liquid chromatography coupled to photodiode array and electrospray ionization mass spectrometry (HPLC/PDA/ESI-MS/MS). A total of 49 phytochemicals were identified in both positive and negative ionization modes. The identified compounds belonged to four classes of secondary metabolites including secoiridoids, flavonoids, pentacyclic triterpenoids and various phenolic compounds. Seasonal variation in chemical composition among the studied cultivars was apparent in autumn and spring. Secologanoside, oleuropein, hydroxy-oleuropein, demethyl oleuropein, gallocatechin, luteolin-O-hexoside, diosmetin, oleanolic acid and maslinic acid were detected in all cultivars in both seasons. Oleuropein-O-deoxyhexoside was tentatively identified for the first time in olive leaf extracts; detected only in the Spanish cultivar Picual (PIC) collected in spring. Also, dihydroxy-oxooleanenoic acid and hydroxy-oxooleanenoic acid, two bioactive pentacyclic triterpenes, were identified. Principle component analysis (PCA) showed good discrimination among the studied cultivars in terms of their botanical origin. This study is considered the first study for non-targeted metabolic profiling of different olive leaf cultivars cultivated in Egypt.

Comprehensive metabolite profiling of Phoenix rupicola pulp and seeds using UPLC-ESI-MS/MS and evaluation of their estrogenic activity in ovariectomized rat model.

Dates have been consumed since ancient times as functional foods which beside their high nutritional value possess various biological activities. Phoenix rupicola T. Anderson (Cliff date palm) produces non-conventional edible dates, however, due to low natural abundance, these dates aren't commercially important as the dates of Phoenix dactylifera L. The present study was designed to evaluate the phytochemical constituents as well as the estrogenic activity of P. rupicola dates. UPLC-ESI-MS/MS approach was used to study the metabolite profile of the 70% aqueous methanol extracts of P. rupicola dates (pulps and seeds) for the first time. A total of fifty-five metabolites were tentatively identified in both extracts, belonging to different classes, chiefly phenolic compounds viz. procyanidins, flavonoid glycosides, hydroxycinnamic acid derivatives, as well as, fatty acids, organic acids and sphingolipids. Acute toxicity studies revealed that the studied extracts were safe at oral doses up to 2 g/kg. Besides, they possessed significant (P < 0.05) estrogenic activity in ovariectomized rat model, as compared to ovariectomized (OVX) and reference standard (17ß-estradiol; OVX-E) groups. Moreover, the extracts showed significant improvement on bone metabolism, lipid profile, liver and kidney functions. In silico docking study revealed that various metabolites possessed high binding affinities to both ERs, where 2-palmitoyl glycerol (-10.28 Kcal/mol) and aminotetradecanetriol (-9.61 Kcal/mol) showed the strongest affinities to Erα and Erß, respectively. Thus, it can be concluded that P. rupicola pulp and seeds possess bioactive phytoconstituents comparable to those in P. dactylifera and can be used as a safe and efficient natural estrogen substitute in postmenopausal women.

Protective effects of Brownea grandiceps (Jacq.) against ϒ-radiation-induced enteritis in rats in relation to its secondary metabolome fingerprint.

Radiation enteritis is the most common complication of radiotherapy in patients with pelvic malignancies. Thus, the radioprotective activity of the total hydro-alcoholic extract (BGE) and the ethyl acetate soluble fraction (EAF) of Brownea grandiceps leaves was evaluated against ϒ-radiation-induced enteritis in rats. (BGE) and (EAF) were characterized using HPLC-PDA-ESI-MS/MS analysis. The total phenolic and flavonoid contents were also quantified. In vivo administration of (BGE) (400 mg/kg) and (EAF) (200 & 400 mg/kg) prevented intestinal injury and maintained the mucosal integrity of irradiated rats through increasing villi length and promoting crypt regeneration. Also, (EAF) showed more potent antioxidant activity than (BGE) through reduction of MDA level and enhancement of GSH content and catalase enzyme activity. (BGE) and (EAF) down-regulated intestinal NF-κB expression leading to diminished expression of downstream inflammatory cytokine TNF-α. Moreover, (EAF) markedly reduced the expression of profibrotic marker TGF-ß1. Seventy-nine compounds were tentatively identified, including flavonoids, proanthocyanidins, polar lipids and phenolic acids. (EAF) showed significantly higher total phenolic and flavonoid contents, as compared to (BGE). Results revealed remarkable radioprotective activity of (BGE) and (EAF), with significantly higher activity for (EAF). The chemical constituents of (BGE) and (EAF) strongly supported their radioprotective activity. To the best of our knowledge, the present study describes for the first time the radioprotective activity of B. grandiceps leaves in relation to its secondary metabolome fingerprint; emphasizing the great promise of B. grandiceps leaves, especially (EAF), to be used as natural radio-protective agent.

Advances in the quality control of fenugreek seeds using chromatographic, spectroscopic and DNA-based techniques: A comprehensive review.

INTRODUCTION: Fenugreek has been used in traditional remedies since ancient times. It has a long history of use against medical ailments as an antidiabetic, anticarcinogenic, hypocholesterolemic, antioxidant, antibacterial, hypoglycemic, gastric stimulant, and anti-anorexia agent. The major active constituents include alkaloids, fibres, saponins, proteins, and amino acids. OBJECTIVES: To provide a comprehensive overview of the application of chromatographic and spectroscopic methods, in addition to DNA-profiling methods to assess the quality of fenugreek. Also, to highlight the recent application of chemometrics combined with quality control methods during the last two decades. METHODOLOGY: A literature search conducted from January 2000 up to December 2020 using various scientific databases (e.g., Scopus, Medline, PubMed, EBSCO, JSTOR, ScienceDirect, Google Scholar, Web of Science and Egyptian Knowledge Bank, Academic Journals, and Springer Link); general web searches were also undertaken using Google applying some related search terms. Studies involving the application of quality control analyses were classified into three categories according to the conducted analysis method including chromatographic [high-performance liquid chromatography (HPLC), high-performance thin-layer chromatography (HPTLC), and gas chromatography (GC)], spectroscopic [ultraviolet (UV), infrared (IR), and nuclear magnetic resonance (NMR)], and DNA-based markers. RESULTS: This review shed the light on relevant studies covering the past two decades, presenting the application of spectroscopic and chromatographic methods and DNA profiling in the quality control of fenugreek. CONCLUSION: The reviewed chromatographic and spectroscopic methods combined with chemometrics provide a powerful tool that could be applied widely for the quality control of fenugreek.

Quality assessment of leaf extracts of 12 olive cultivars and impact of seasonal variation based on UV spectroscopy and phytochemcial content using multivariate analyses.

INTRODUCTION: Recently, focus has been made on the health-oriented uses of olive leaves, a byproduct of olive production, as a potential source of antioxidants. Oleuropein is one of the phenolic components in olive leaves known for its high antioxidant value. OBJECTIVE: The main aim of the current study was constructing a model for the quality assessment of olive leaves and their potential phytochemical content and hence biological value as well. The phytochemical variation in olive leaves in both flowering (spring) and fruiting seasons (autumn) was also investigated. METHODS: In this study, the leaves of 12 different olive cultivars from different geographical origins growing in Egypt were assessed for their oleuropein content, total flavonoid (TF) content and total polyphenol (Pph) content in spring and autumn via ultraviolet (UV) spectroscopy and high-performance liquid chromatography (HPLC) coupled to multivariate data analyses. The antioxidant activity of olive leaf extracts was assessed using 2,2'-diphenyl-1-picrylhydrazyl (DPPH) assay. RESULTS: Higher levels of oleuropein, TF and Pph content were found in spring with the highest oleuropein content in the Spanish cultivar; Manzanillo, followed by the Italian cultivar Coratina and the Egyptian Agizi Okasi (218.94, 151.58 and 122.18 mg/100 g of dried leaf extract, respectively). UV spectra was also measured and the collected data were coupled to multivariate analyses showing clustering of cultivars with common geographical origin. CONCLUSION: Our findings emphasised the influence of collection time and type of cultivar on the chemical profile of olive leaves. The model presented herein, serves for the quality assessment of olive leaves based on their phytochemical profile.

Quality control and long-term stability study of ginger from different geographical origins using chemometrics.

BACKGROUND: Ginger rhizome (Zingiber officinale) is a well-known spice and medicinal plant that is widely used in the Egyptian market as a spice, flavor and medicinal herb for different diseases. Since it is not cultivated as rhizomes in Egypt, ginger is imported from other countries, which may impact its quality. In this study, UV spectroscopy and high-performance liquid chromatography (HPLC) were applied as efficient available techniques for the discrimination and quality control of ginger collected from different geographical origins in combination with chemometrics. In addition, HPLC was applied to investigate the stability of ginger samples upon storage for 3 years to trace the changes in their main active constituents. RESULTS: Data obtained from both UV and HPLC in combination with Principal Component Analysis (PCA) displayed proper discrimination of the samples according to their geographical origins. Regarding the stability study, ginger samples showed a significant decrease in quality after storage for 3 years, in which significant variation in the main pungent principles (6-, 8-, 10-gingerols and 6-shogaol) were observed. PCA failed to discriminate between ginger samples after long-time storage, so the applied model could discriminate between ginger samples before and after storage. CONCLUSION: UV and HPLC in combination with chemometrics can be applied as a successful tool for the study of quality, stability and geographical discrimination of ginger. © 2020 Society of Chemical Industry.

Comparative metabolome-based classification of Senna drugs: a prospect for phyto-equivalency of its different commercial products.

INTRODUCTION: The demand to develop efficient and reliable analytical methods for the quality control of nutraceuticals is on the rise, together with an increase in the legal requirements for safe and consistent levels of its active principles. OBJECTIVE: To establish a reliable model for the quality control of widely used Senna preparations used as laxatives and assess its phyto-equivalency. METHODS: A comparative metabolomics approach via NMR and MS analyses was employed for the comprehensive measurement of metabolites and analyzed using chemometrics. RESULTS: Under optimized conditions, 30 metabolites were simultaneously identified and quantified including anthraquinones, bianthrones, acetophenones, flavonoid conjugates, naphthalenes, phenolics, and fatty acids. Principal component analysis (PCA) was used to define relative metabolite differences among Senna preparations. Furthermore, quantitative 1H NMR (qHNMR) was employed to assess absolute metabolites levels in preparations. Results revealed that 6-hydroxy musizin or tinnevellin were correlated with active metabolites levels, suggesting the use of either of these naphthalene glycosides as markers for official Senna drugs authentication. CONCLUSION: This study provides the first comparative metabolomics approach utilizing NMR and UPLC-MS to reveal for secondary metabolite compositional differences in Senna preparations that could readily be applied as a reliable quality control model for its analysis.

Phytoconstituents from Polyscias guilfoylei leaves with histamine-release inhibition activity.

Phytochemical investigation of Polyscias guilfoylei leaves extract (PGE) led to the isolation of nine compounds, that is, ent-labda-8(17),13-diene-15,18-diol (1), stigmasterol (2), spinasterol (3), N-(1,3-dihydroxyoctadecan-2-yl) palmitamide (4), panaxydiol (5), 3-O-ß-d-glucopyranosylstigmasta-5,22-diene-3-ß-ol (6), (8Z)-2-(2 hydroxypentacosanoylamino) octadeca-8-ene-1,3,4-triol (7), 4-hydroxybenzoic acid (8), and tamarixetin 3,7-di-O-α-L-rhamnopyranoside (9). Compound 4 is reported in this study for the first time in nature whereas compound 9 is reported for the second time. Structural elucidation of the compounds was carried out using Nuclear Magnetic Resonance and Electrospray Ionization coupled with Mass Spectrometry spectroscopic analyses. PGE and compounds 4 and 9 exhibited weak cytotoxicity against both MCF-7 and HCT-116 cell lines using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay. The antimicrobial activity of PGE and compounds 4 and 9 was evaluated using the agar diffusion method. Escherichia coli was the most susceptible Gram-negative bacteria toward PGE with a minimum inhibitory concentration value of 9.76 µg/mL, whereas compounds 4 and 9 did not show any antimicrobial activity. Compound 4 exhibited promising inhibition of histamine release using U937 human monocytes with an IC50 value of 38.65 µg/mL.

Curcumin or bisdemethoxycurcumin for nose-to-brain treatment of Alzheimer disease? A bio/chemo-informatics case study.

The current study introduces a new idea of utilising several bio/chemoinformatics tools in comparing two bio-similar natural molecules viz. curcumin and bisdemethoxycurcumin (BDMC) in order to select a potential nose-to-brain remedy for Alzheimer disease. The comparison comprised several bio/chemo informatics tools. It encompassed all levels starting from loading the drug in a certain carrier; PLGA nanoparticles, to the biopharmaceutical level investigating the interaction with mucin and inhibition of P-gp blood-brain barrier efflux pumps. Finally, the therapeutic level was investigated by studying the interaction with pharmacological targets such as amyloid peptide plaques and cyclooxygenase2 enzyme responsible for the inflammatory reactions of the studied disease. The comparison revealed the superiority of curcumin over BDMC. Five new analogues were also hypothesised where diethoxybisdemethoxycurcumin was  recommended as a superior molecule. This work introduced the virtual utilisation of bio/chemo informatics tools as a reliable and economic alternative to the exhausting and resources-consuming wet-lab experimentation.

Selecting optimum protein nano-carriers for natural polyphenols using chemoinformatics tools.

BACKGROUND: The normal fate of any natural product with a therapeutic potential is to be formulated into an effective medicine. However, the conventional methods of selecting the suitable formulations or carriers based on the formulator experiences, trials and errors as well as materials availability do not usually yield the optimal results. HYPOTHESIS: We hypothesize the possibility of the virtual optimum selection of a protein carrier for two polyphenolic compounds widely investigated for their chemopreventive effects; resveratrol and curcumin using a combination of some chemoinformatics tools. METHODS: Two protein-based nanoparticles namely; albumin and gelatin nanoparticles were compared as carriers for the two selected phytochemicals; resveratrol and curcumin. Resveratrol-albumin, resveratrol-gelatin and curcumin-albumin results were gathered from the literature. While, a new combination (formulation), comprising curcumin as the cargo and gelatin nanoparticles as the carrier, was prepared and evaluated as a potential medicine for breast cancer. Combined chemoinformatics tools, namely; molecular dynamics and molecular docking were used to determine the optimum carrier for each of the two chemopreventive agents. RESULTS: A new curcumin-gelatin nanoparticulate formulation was prepared and proven cytotoxic after an application period of 48h on MCF-7 breast cancer cell-lines scoring an IC50 value of 64.8µg/ml. The utilized chemoinformatics tools comprising the molecular dynamics simulations of the protein nano-particulate drug-carriers followed by the molecular docking of phytochemical drugs on these carriers could capture the optimum protein carrier for each of the tested phytochemical and hence propose a successful formulation. CONCLUSION: This study presents one in a series that proves the novel addressed concept of the utilization of computational tools rather than wet-lab experimentation in providing better selection of drug-carrier pairs aiming for better formulations and the subsequent successful therapeutic effects.

Anti-infective and cytotoxic properties of Bupleurum marginatum.

BACKGROUND: Bupleurum marginatum Wall. ex DC (Apiaceae) is a perennial herb widely used in traditional Chinese and Kampo medicine for the treatment of various infectious diseases. The biological activities of B. marginatum have not been fully investigated. This study aims to investigate the antitrypanosomal, antimicrobial and antiviral activities of methanol (ME) and dichloromethane (DCM) extracts of B. marginatum aerial parts and the ability of both extracts to inhibit the growth of different cancer cell lines. METHODS: Phytochemical characterization of the extracts was performed by LC-MS profiling. The antitrypanosomal activity was evaluated using the resazurin method. The antimicrobial activity was assessed using agar diffusion and microdilution methods, and the minimum inhibitory concentration (MIC) values were determined. The antiviral activity was determined for 6.25, 12.5, and 50 µg/mL doses using a plaque reduction assay. Cytotoxicity was investigated in eight cancer cell lines (Caco-2, CCL-81, CCRF-CEM, COS-7, HL-60, MIA PaCa-2, MCF-7, and PANC-1) using the MTT assay and the caspase 3/7 activity was determined over the range of 62.5-1000 µg/mL. RESULTS: Phytochemical analyses resulted in the characterization of 15 components, mainly flavonoids and lignans. The DCM extract showed significant antitrypanosomal activity (IC50: 36.21 µg/mL) and moderate activity against Streptococcus pyogenes (MIC value: 0.25 mg/mL). At a dose of 12.5 µg/mL, the DCM extract inhibited 73.6% of the plaque production by hepatitis A virus. CCRF-CEM cells were the most sensitive to both extracts (IC50: 12.5-22.7 µg/mL). The cytotoxicity was mediated by induction of apoptosis (19-fold increase in the cellular caspase 3/7 level after treatment with the DCM extract at 1 mg/mL). CONCLUSIONS: ME and DCM extract of B. marginatum showed anti-infective and antiproliferative effects.

Metabolomics driven analysis of artichoke leaf and its commercial products via UHPLC-q-TOF-MS and chemometrics.

The demand to develop efficient and reliable analytical methods for the quality control of herbal medicines and nutraceuticals is on the rise, together with an increase in the legal requirements for safe and consistent levels of active principles. Here, we describe an ultra-high performance liquid chromatography method (UHPLC) coupled with quadrupole high resolution time of flight mass spectrometry (qTOF-MS) analysis for the comprehensive measurement of metabolites from three Cynara scolymus (artichoke) cultivars: American Green Globe, French Hyrious, and Egyptian Baladi. Under optimized conditions, 50 metabolites were simultaneously quantified and identified including: eight caffeic acid derivatives, six saponins, 12 flavonoids and 10 fatty acids. Principal component analysis (PCA) was used to define both similarities and differences among the three artichoke leaf cultivars. In addition, batches from seven commercially available artichoke market products were analysed and showed variable quality, particularly in caffeic acid derivatives, flavonoid and fatty acid contents. PCA analysis was able to discriminate between various preparations, including differentiation between various batches from the same supplier. To the best of our knowledge, this study provides the first approach utilizing UHPLC-MS based metabolite fingerprinting to reveal secondary metabolite compositional differences in artichoke leaf extracts.

A modern approach to the authentication and quality assessment of thyme using UV spectroscopy and chemometric analysis.

INTRODUCTION: Recently, the fields of chemometrics and multivariate analysis have been widely implemented in the quality control of herbal drugs to produce precise results, which is crucial in the field of medicine. Thyme represents an essential medicinal herb that is constantly adulterated due to its resemblance to many other plants with similar organoleptic properties. OBJECTIVE: To establish a simple model for the quality assessment of Thymus species using UV spectroscopy together with known chemometric techniques. The success of this model may also serve as a technique for the quality control of other herbal drugs. MATERIALS AND METHODS: The model was constructed using 30 samples of authenticated Thymus vulgaris and challenged with 20 samples of different botanical origins. The methanolic extracts of all samples were assessed using UV spectroscopy together with chemometric techniques: principal component analysis (PCA), soft independent modeling of class analogy (SIMCA) and hierarchical cluster analysis (HCA). RESULTS: The model was able to discriminate T. vulgaris from other Thymus, Satureja, Origanum, Plectranthus and Eriocephalus species, all traded in the Egyptian market as different types of thyme. The model was also able to classify closely related species in clusters using PCA and HCA. The model was finally used to classify 12 commercial thyme varieties into clusters of species incorporated in the model as thyme or non-thyme. CONCLUSION: The model constructed is highly recommended as a simple and efficient method for distinguishing T. vulgaris from other related species as well as the classification of marketed herbs as thyme or non-thyme.

Application of chemometrics in authentication of herbal medicines: a review.

INTRODUCTION: Herbal medicines (HM) and their preparations have been widely used for hundreds of years all over the world. However, they have not been officially recognised due to a lack of adequate or accepted research methodology for their evaluation. OBJECTIVE: To present a concise overview of the recent applications of chemometrics in solving the ambiguity of herbal medicine authentication during the last two decades. METHODOLOGY: Studies involving the applications of chemometric analysis in combination with different analytical methods were classified according to the method of analysis used including chromatographic (HPLC, GC and CE), spectroscopic (NMR, IR, UV and ICP) and genetic analysis (RAPD). The purpose of each of these studies was classified into one of three main categories: taxonomic discrimination, quality assessment or classification between plants of different geographic origins. RESULTS: This review comprises over 150 studies, covering the past two decades, emphasising the significance of chemometric methods in the discrimination of many herbs from closely related species and from adulterants, based on the principal bioactive components and phytochemical diversity. Furthermore, the differentiation between varieties and hybrids was achieved in addition to the prediction of the active components by quantitative methods of analysis. Discrimination according to geographical origin and localities, processing methods, DNA profiling and metabolomics were also efficiently investigated. CONCLUSION: Chemometric methods have provided an efficient and powerful tool for the quality control and authentication of different herbs.