GC-MS ANALYSIS OF BIOACTIVE COMPOUNDS IN ETHANOL LEAVES EXTRACT OF SPHENOCENTRUM JOLLYANUM AND THEIR BIOLOGICAL ACTIVITIES.

Sphenocentrum jollyanum is a plant genus of the family Menispermaceae. It has high medicinal importance as it is used traditionally to treat various diseases such as jaundice, breast engorgement related to the menstrual cycle, tumour, fibroids and improve the health of people. The present investigation was carried out to analyze the bioactive compounds present in ethanol crude extract of Sphenocentrum jollyanum leaves using GC-MS analysis. GC-MS analysis of ethanol extract Sphenocentrum jollyanum was done using a 7890A GC system (Agilent Technologies), coupled with 5977B MSD (Agilent Technologies) while the mass spectra of the compounds found in the extract was matched with the National Institute of Standards and Technology (NIST) library. A total of 45 bioactive compounds representing 99.98% of the total extract based on the retention time, peak area, molecular formula, molecular weight, and biological activities were identified by GC-MS which ranges from high molecular weight to low molecular weight compounds. The major compounds identified with their peak area percentages were 2,4-Di-tertbutylphenol, (21.05%), Z-8-Methyl-9-tetradecenoic acid (19.12), Hexadecanoic acid, ethyl ester (7.86%), Diisooctyl phthalate (7.13%), Phytol, Oleic Acid (7.03), 6,9,12- Octadecatrien-1-ol (6.65%), 3-Eicosene, (E)-(4.63%), 2-Methyl-Z, Z-3,13-octadecadienol (4.24%), n- Hexadecanoic acid (4.09%), trans-13-Octadecenoic acid (3.81%), Cyclohexene, 4-(4-ethylcyclohexyl) -1- pentyl- (3.74%), Dibutyl phthalate (3.20%), and 9-Oxabicyclo (6.1.0) nonane, cis-(3.18%). The presence of these major phytoconstituents in the leaf extract provides various biological activities including antifungal, antibacterial, antioxidant, anti-inflammatory, and anti-tumour which supports the ethno-medicinal uses of the plant in curing diseases. We recommend

Effects of Delivering Guanidinoacetic Acid or Its Prodrug to the Neural Tissue: Possible Relevance for Creatine Transporter Deficiency

The creatine precursor guanidinoacetate (GAA) was used as a dietary supplement in humans with no adverse events. Nevertheless, it has been suggested that GAA is epileptogenic or toxic to the nervous system. However, increased GAA content in rodents affected by guanidinoacetate methyltransferase (GAMT) deficiency might be responsible for their spared muscle function. Given these conflicting data, and lacking experimental evidence, we investigated whether GAA affected synaptic transmission in brain hippocampal slices. Incubation with 11.5 μM GAA (the highest concentration in the cerebrospinal fluid of GAMT-deficient patients) did not change the postsynaptic compound action potential. Even 1 or 2 mM had no effect, while 4 mM caused a reversible decrease in the potential. Guanidinoacetate increased creatine and phosphocreatine, but not after blocking the creatine transporter (also used by GAA). In an attempt to allow the brain delivery of GAA when there was a creatine transporter deficiency, we synthesized diacetyl guanidinoacetic acid ethyl ester (diacetyl-GAAE), a lipophilic derivative. In brain slices, 0.1 mM did not cause electrophysiological changes and improved tissue viability after blockage of the creatine transporter. However, diacetyl-GAAE did not increase creatine nor phosphocreatine in brain slices after blockage of the creatine transporter. We conclude that: (1) upon acute administration, GAA is neither epileptogenic nor neurotoxic; (2) Diacetyl-GAAE improves tissue viability after blockage of the creatine transporter but not through an increase in creatine or phosphocreatine. Diacetyl-GAAE might give rise to a GAA–phosphoGAA system that vicariates the missing creatine–phosphocreatine system. Our in vitro data show that GAA supplementation may be safe in the short term, and that a lipophilic GAA prodrug may be useful in creatine transporter deficiency.

(E)-9-Octadecenoic Acid Ethyl Ester Derived from Lotus Seedpod Ameliorates Inflammatory Responses by Regulating MAPKs and NF-κB Signalling Pathways in LPS-Induced RAW264.7 Macrophages

Inflammation is generally considered a key risk factor in the progress of several chronic diseases, such as arthritis, gastritis, and hepatitis. Natural products with anti-inflammatory ability have played a great role in the process of overcoming these inflammatory diseases. In this study, we evaluated the anti-inflammatory activities of ten natural compounds derived from lotus seedpod and discovered (E)-9-octadecenoic acid ethyl ester (E9OAEE) inhibited the production of nitric oxide (NO) optimally in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Furthermore, we explored the effects of E9OAEE on inflammatory responses and the underlying mechanisms in LPS-induced RAW264.7 macrophages. The results indicated that E9OAEE significantly suppressed the production of NO, prostaglandin E2 (PGE2), and tumour necrosis factor-α (TNFα) in a dose-dependent manner. The protein expression and mRNA levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2) were inhibited by pretreatment of E9OAEE. Furthermore, E9OAEE restrained the phosphorylation of mitogen-activated protein kinase (MAPKs) family members, ERK, P38, and JNK stimulated by LPS-treated for 30 min and prevented the nuclear translocation of nuclear factor-kappa B (NF-κB) prompted by LPS-treated for 6 h in RAW264.7 macrophages. Taken together, we discovered an anti-inflammatory component from lotus seedpod and identified E9OAEE attenuated the inflammatory response in LPS-induced RAW264.7 macrophages probably by regulating the activation of MAPKs and NF-κB signalling pathways, which would provide some base for the development of new anti-inflammatory drugs.

GC-MS Analysis and Biomedical Therapy of Oil from n-Hexane Fraction of Scutellaria edelbergii Rech. f.: In Vitro, In Vivo, and In Silico Approach

The current study aimed to explore the crude oils obtained from the n-hexane fraction of Scutellaria edelbergii and further analyzed, for the first time, for their chemical composition, in vitro antibacterial, antifungal, antioxidant, antidiabetic, and in vivo anti-inflammatory, and analgesic activities. For the phytochemical composition, the oils proceeded to gas chromatography-mass spectrometry (GC-MS) analysis and from the resultant chromatogram, 42 bioactive constituents were identified. Among them, the major components were linoleic acid ethyl ester (19.67%) followed by ethyl oleate (18.45%), linolenic acid methyl ester (11.67%), and palmitic acid ethyl ester (11.01%). Tetrazolium 96-well plate MTT assay and agar-well diffusion methods were used to evaluate the isolated oil for its minimum inhibitory concentrations (MIC), minimum bactericidal concentration (MBC), half-maximal inhibitory concentrations (IC50), and zone of inhibitions that could determine the potential antimicrobial efficacy’s. Substantial antibacterial activities were observed against the clinical isolates comprising of three Gram-negative bacteria, viz., Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa, and one Gram-positive bacterial strain, Enterococcus faecalis. The oils were also effective against Candida albicans and Fusarium oxysporum when evaluated for their antifungal potential. Moreover, significant antioxidant potential with IC50 values of 136.4 and 161.5 µg/mL for extracted oil was evaluated through DPPH (1,1-Diphenyl-2-picryl-hydrazyl) and ABTS assays compared with standard ascorbic acid where the IC50 values were 44.49 and 67.78 µg/mL, respectively, against the tested free radicals. The oils was also potent, inhibiting the α-glucosidase (IC50 5.45 ± 0.42 µg/mL) enzyme compared to the standard. Anti-glucosidase potential was visualized through molecular docking simulations where ten compounds of the oil were found to be the leading inhibitors of the selected enzyme based on interactions, binding energy, and binding affinity. The oil was found to be an effective anti-inflammatory (61%) agent compared with diclofenac sodium (70.92%) via the carrageenan-induced assay. An appreciable (48.28%) analgesic activity in correlation with the standard aspirin was observed through the acetic acid-induced writhing bioassay. The oil from the n-hexane fraction of S. edelbergii contained valuable bioactive constituents that can act as in vitro biological and in vivo pharmacological agents. However, further studies are needed to uncover individual responsible compounds of the observed biological potentials which would be helpful in devising novel drugs.

Chemoenzymatic Stereodivergent Synthesis of All the Possible Stereoisomers of the 2,3-Dimethylglyceric Acid Ethyl Ester

2,3-dihydroxy-2-methylbutyric acid, also known as 2,3-dimethylglyceric acid, constitutes the acyl and/or the alcoholic moiety of many bioactive natural esters. Herein, we describe a chemoenzymatic methodology which gives access to all the four possible stereoisomers of the 2,3-dimethylglyceric acid ethyl ester. The racemic ethyl α-acetolactate, produced by the N-heterocycle carbene (NHC)-catalyzed coupling of ethyl pyruvate and methylacetoin was employed as the starting material. The racemic mixture was resolved through (S)-selective reductions, promoted by the acetylacetoin reductase (AAR) affording the resulting ethyl (2R,3S)-2,3-dimethylglycerate; the isolated remaining (S)-ethyl α-acetolactate was successively treated with baker’s yeast to obtain the corresponding (2S,3S) stereoisomer. syn-2,3-Dimethylgliceric acid ethyl ester afforded by reducing the rac-α-acetolactate with NaBH4 in the presence of ZnCl2 was kinetically resolved through selective acetylation with lipase B from Candida antarctica (CAL-B) and vinyl acetate to access to (2S,3R) stereoisomer. Finally, the (2R,3R) stereoisomer, was prepared by C3 epimerization of the (2R,3S) stereoisomer recovered from the above kinetic resolution, achieved through the TEMPO-mediated oxidation, followed by the reduction of the produced ketone with NaBH4. The resulting 2,3-dimethylglycertate enriched in the (2R,3R) stereoisomer was submitted to stereospecicific acetylation with vinyl acetate and CAL-B in order to separate the major stereoisomer. The entire procedure enabled conversion of the racemic α-acetolactate into the four enantiopure stereoisomers of the ethyl 2,3-dihydroxy-2-methylbutyrate with the following overall yields: 42% for the (2R,3S), 40% for the (2S,3S), 42% for the (2S,3R) and 20% for the (2R,3R).

Dietary Amylose/Amylopectin Ratio Modulates Cecal Microbiota and Metabolites in Weaned Goats

Increasing the ratio of amylose in the diet can increase the quantity of starch that flows to the large intestine for microbial fermentation. This leads to the alteration of microbiota and metabolite of the hindgut, where the underlying mechanism is not clearly understood. The present study used a combination of 16S amplicon sequencing technology and metabolomics technique to reveal the effects of increasing ratios of amylose/amylopectin on cecal mucosa- and digesta-associated microbiota and their metabolites in young goats. Twenty-seven Xiangdong black female goats with average body weights (9.00 ± 1.12 kg) were used in this study. The goats were randomly allocated to one of the three diets containing starch with 0% amylose corn (T1), 50% high amylose corn (T2), and 100% high amylose corn (T3) for 35 days. Results showed that cecal valerate concentration was higher (P < 0.05) in the T2 group than those in the T1 and T3 groups. The levels of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were decreased (P < 0.05) in cecal tissue while IL-10 was increased (P < 0.05) in the T2 group when compared with T1 or T3 groups. At the phylum level, the proportion of mucosa-associated Spirochaetes was increased (P < 0.05), while Proteobacteria was deceased by feeding high amylose ratios (P < 0.05). The abundance of Verrucomicrobia was decreased (P < 0.05) in the T3 group compared with the T1 and T2 groups. The abundance of digesta-associated Firmicutes was increased (P < 0.05) while Verrucomicrobia and Tenericutes were deceased (P < 0.05) with the increment of amylose/amylopectin ratios. The LEfSe analysis showed that a diet with 50% high amylose enriched the abundance of beneficial bacteria such as Faecalibacterium and Lactobacillus in the digesta and Akkermansia in the mucosa compared with the T1 diet. The metabolomics results revealed that feeding a diet containing 50% high amylose decreased the concentration of fatty acyls-related metabolites, including dodecanedioic acid, heptadecanoic acid, and stearidonic acid ethyl ester compared with the T1 diet. The results suggested that a diet consisting of 50% high amylose could maintain a better cecal microbiota composition and host immune function.

Exploring the mechanism of Yixinyin for myocardial infarction by weighted co-expression network and molecular docking

Yixinyin, the traditional Chinese medicine, has the effects of replenishing righteous qi, and promoting blood circulation to eliminate blood stagnation. It is often used to treat patients with acute myocardial infarction (MI). The purpose of our study is to explore the key components and targets of Yixinyin in the treatment of MI. In this study, we analyzed gene expression data and clinical information from 248 samples of MI patients with the GSE34198, GSE29111 and GSE66360 data sets. By constructing a weighted gene co-expression network, gene modules related to myocardial infarction are obtained. These modules can be mapped in Yixinyin PPI network. By integrating differential genes of healthy/MI and unstable angina/MI, key targets of Yixinyin for the treatment of myocardial infarction were screened. We validated the key objectives with external data sets. GSEA analysis is used to identify the biological processes involved in key targets. Through molecular docking screening, active components that can combine with key targets in Yixinyin were obtained. In the treatment of myocardial infarction, we have obtained key targets of Yixinyin, which are ALDH2, C5AR1, FOS, IL1B, TLR2, TXNRD1. External data sets prove that they behave differently in the healthy and MI (P < 0.05). GSEA enrichment analysis revealed that they are mainly involved in pathways associated with myocardial infarction, such as viral myocarditis, VEGF signaling pathway and type I diabetes mellitus. The docking results showed that the components that can be combined with key targets in YixinYin are Supraene, Prostaglandin B1, isomucronulatol-7,2′-di-O-glucosiole, angusifolin B, Linolenic acid ethyl ester, and Mandenol. For that matter, they may be active ingredients of Yixinyin in treating MI. These findings provide a basis for the preliminary research of myocardial infarction therapy in traditional Chinese medicine and provide ideas for the design of related drugs.

Phytochemical Screening and Identification of Bioactive Constituents of the Chloroform Extract of Icacina trichantha Tuber Peel Oliv

This study was carried out to identify the bioactive constituents of the tuber peels of Icacina trichanthalinna of the Icacinales family. This plant is basically selected due to its widespread use in herbal treatment. Nine compounds were identified in the chloroform extract of the tuber peel. Undecane (43.254%),2-hexanone (23.299%), Dodecanoic acid ethyl ester (6.244%) and 9-octadecynoic acid (5.915%) appeared to be the most prevailing compounds and they are known to possess antibacterial, antiviral, antioxidant, antimycobacterial, hypercholesterolemic activities. Also identified was triarachine which has been reported to have some biological activity.

Evaluation of the bioactive compounds of Vernonia amygdalina Delile extracts and their antibacterial potentials on water-related bacteria

Background This study focused on the evaluation of the bioactive compounds of Vernonia amygdalina Del. leaf extracts and their antibacterial potential on some water-related bacterial isolates. The bacterial isolates were confirmed using standard microbiological test. The leaves of V. amygdalina were subjected to extraction using the Maceration method with water and ethanol as the extraction solvents. Gas chromatography–mass spectroscopy (GC–MS) was carried out on extracts. Antibacterial susceptibility test of V. amygdalina extracts on isolates was carried out. Results Aqueous extract of V. amygdalina had a higher percentage yield (11.89%) than the ethanol extract (5.37%). The GC–MS carried out revealed the presence of butanoic acid, squalene, palmitaldehyde, octadecanoic acid, Z-hexadecanoic acid ethyl ester, oxirane, tetradecyl, 3- methyl-2-phenylindole, n-heneicosane, phytol, methyl-2-O-benzyl-d-arabinofuranoside, cholest-5-en-3-ol acetate; with hexadecanoic acid ethyl ester and 1,1-diethoxy-3methylbutane having the highest percentage composition of 24.37% and 13.42% in aqueous and ethanol extract, respectively, aqueous extract highly inhibited Escherichia coli with an inhibition zone of 10.333 ± 0.882 and 36.667 ± 0.882 for 25 mg/ml and 100 mg/ml, respectively, while the ethanol extracts inhibited most of the isolates with an inhibition range of 7.000 ± 1.155 to 30.333 ± 0.882. The minimum inhibitory concentration for both extracts on the isolates varies from 25 to 50 mg/ml. Conclusions The ethanol extract of V. amygdalina had a higher inhibitory activity on the bacterial isolates than water. These findings indicate the potential of ethanol extract of V. amygdalina leaf in the treatment of water borne infections.

Sorghum extract: Phytochemical, proximate, and GC-MS analyses

Introduction. Sorghum is available cereal seeds of African origin belonging to the Poaceae family. However, its metabolites and proximate composition have not studied well, which led to the under-utilization of this cereal. This research aimed to investigate the classes of phytochemical and proximate compositions of sorghum extract in order to assess its nutraceutical potential for food chemistry and dietary formulations. Study objects and methods. We studied the sorghum seed oil extract obtained with the help of a Soxhlet extractor. Sorghum was purchased in Ota, Nigeria. The bioactive compounds were identified by standard methods of phytochemical screening, the nutritional content was investigated with proximate analysis, and the secondary metabolites in the sorghum extract were determined using gas chromatography – mass spectrometry (GC-MS). Result and discussion. The phytochemical screening showed the presence of steroids, saponins, terpenoids, alkaloids, cardiac glycosides, and quinones in the sorghum extract. The oil yield obtained was 11.00 ± 0.18%. The proximate analysis revealed 5.94% moisture content, 3.05% ash, 0.20% crude fiber, 11.00% fat, 5.54% protein, and 74.27% carbohydrates. The selected physicochemical parameters measured in the sorghum extract included cloud point (0.40°C), specific gravity at 25°C (0.81), and refractive index (1.46). The GC-MS analysis revealed the presence of 9,12-octadecadienoic acid (Z,Z)-, stigmasterol, 8-dodecen-1-ol, acetate, (Z)-, vitamin E, linoleic acid ethyl ester, and 9,12-octadecadienoic acid, methyl ester, which accounted for about 85% in the sorghum composition. Other constituents, presented at lower amounts, included 12-heptadecyn-1-ol, 1H-Imidazole-5-ethanamine, 1-methyl-, and cyclononene. Conclusion. The findings of this study revealed high nutritive potential of sorghum, which make it a rich source of energy for humans and animals.