Crude Extract Of Stem Bark Research Articles

Molecular Networking-Based Metabolome, In Vitro Antidiabetic and Anti-Inflammatory Effects of Breonadia salicina (Vahl) Hepper & J.R.I. Wood.

Breonadia salicina (Vahl) Hepper & J.R.I. Wood is widely distributed throughout Africa. It is used ethnobotanically to treat various diseases. However, the metabolic profile of the Breonadia species is not well characterized and the metabolites that are responsible for the bioactivity of this plant remain unknown. Therefore, there is a need to determine the phytochemical and bioactivity profile to identify metabolites that contribute to the antidiabetic, anti-inflammatory and antiproliferation activity, including the genotoxicity and cytotoxic effects, of Breonadia salicina. The study is aimed at exploring the metabolomic profile antidiabetic, anti-inflammatory and antiproliferation activity, as well as the genotoxicity and cytotoxicity effects, of constituents of B. salicina. The compounds in the B. salicina extract were analyzed by ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS), and the resultant data were further analyzed using a molecular networking approach. The crude stem bark and root extracts showed the highest antidiabetic activity against α-amylase at the lowest test concentration of 62.5 µg/mL, with 74.53 ± 0.74% and 79.1 ± 1.5% inhibition, respectively. However, the crude stem bark and root extracts showed the highest antidiabetic activity against α-glucosidase at the lowest test concentration of 31.3 µg/mL, with 98.20 ± 0.15% and 97.98 ± 0.22% inhibition, respectively. The crude methanol leaf extract showed a decrease in the nitrite concentration at the highest concentration of 200 µg/mL, with cell viability of 90.34 ± 2.21%, thus showing anti-inflammatory activity. No samples showed significant cytotoxic effects at a concentration of 10 µg/mL against HeLa cells. Furthermore, a molecular network of Breonadia species using UPLC-QTOF-MS with negative mode electrospray ionization showed the presence of organic oxygen compounds, lipids, benzenoids, phenylpropanoids and polyketides. These compound classes were differentially distributed in the three different plant parts, indicating the chemical differences between the stem bark, root and leaf extracts of B. salicina. Therefore, the identified compounds may contribute to the antidiabetic and anti-inflammatory activity of Breonadia salicina. The stem bark, root and leaf extracts of B. salicina yielded thirteen compounds identified for the first time in this plant, offering a promising avenue for the discovery of new lead drugs for the treatment of diabetes and inflammation. The use of molecular networking produced a detailed phytochemical overview of this Breonadia species. The results reported in this study show the importance of searching for bioactive compounds from Breonadia salicina and provide new insights into the phytochemical characterization and bioactivity of different plant parts of Breonadia salicina.

Evaluation of Alchornea cordifolia Extracts for Larvicidal Activity against Anopheles Mosquitoes: Implications for Malaria Control

Alchornea cordifolia is a botanical specimen renowned for its medicinal benefits. The phytochemicals present in this plant boast antioxidant, anti-inflammatory, antimicrobial, and analgesic properties, making it a versatile and effective remedy. This plant has been utilized for several decades in treating various ailments like; malaria, diarrhea, and respiratory infections. To evaluate its efficacy in eradicating mosquito larvae, extractions from both the plant's bark and leaves were obtained using various methods, including crude methods, hot water, and ethanol. The plant was macerated in alcohol and hot water, while the bioassay was carried out using a static non-renewal test. The results of the bioassay indicated that the LC50 values for the various extracts of the plant. The ethanol extract was the most active against the mosquito larvae, with an LC50 value of 8.37 ppm. It was followed by the crude stem-bark extract, which had an LC50 value of 13.18 ppm, and the hot water extract, which had an LC50 value of 110.00 ppm. The plant's diverse range of phytochemicals may have played a role in its potency in eliminating mosquito larvae. This activity of the plant strongly suggests that Alchornea cordifolia has insecticidal properties as a larvicide against malaria.

Acute toxicity profile of crude methanolic stem bark extract of Parkia biglobosa in West African Dwarf (WAD) goats

Despite the high demand for herbal medicines for treatment of health disorder, there are still concerns associated with not only their use, but their safety. This study was aimed at providing information on the potential toxicity profile of the stem bark extracts of P. biglobosa in goats. For this purpose, the stem bark parts of P. biglobosa were collected and processed. The acute toxicity, biochemical and histopathological analysis in goats were determined. The results of phytochemical constituents revealed the presence of alkaloids, anthraquinones, cardiac glycosides, glycosides, oils, phlobatannins, reducing sugar, saponins and tannins (condensed and hydrolysable). The results of acute toxicity studies showed no mortality with up to 5000 mg/ kg of the body weight. The blood collected revealed that, CMSBE has no significant (p>0.05) effect on biochemical parameters of WAD goats. However, there were slight changes in glucose concentration, cholesterol and triglyceride level, though they were no statistically significant (p>0.05). The extracts did not also have deleterious effect on morphology of the tissues observed during histopathology. The study has scientifically established that, stem bark extract of P. biglobosa have LD50 that is greater than 5,000 mg/kg and it can be inferred that the plant might have a wide margin of safety typical of treatment of some diseases. It is therefore recommended that despite the non-toxicity as demonstrated in this present study, detailed toxic and pharmacological effects of the extracts of P. biglobosa stem bark in other animals should be investigated.

Effects of seasonal variation on phytochemicals contributing to the antimalarial and antitrypanosomal activities of Breonadia salicina using a metabolomic approach

This study involves the investigation of various plant parts of Breonadia salicina (Vahl) Hepper and J.R.I. Wood across multiple consecutive seasons. It aims to delve into the phytochemistry of these different plant parts and establish connections between the findings and their biological activities. This comprehensive approach employs metabolomics techniques, with the ultimate goal of exploring the potential for drug development. Samples were collected in Fondwe, a village in Limpopo (South Africa), based on local reports of the efficacy of this plant used by traditional healers in the area. The antimalarial and antitrypanosomal activities of samples collected over the seasons were determined with the parasite lactate dehydrogenase (pLDH) and specific Trypanosoma brucei assays, respectively. Consequently, a total of 24 compounds were tentatively identified through ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Chemical profiles of the different plant parts of Breonadia salicina collected in different seasons produced contrasting metabolic profiles. Chemometric analysis of the UPLC-QTOF-MS data enabled us to determine the chemical variability of the crude stem bark, root and leaf extracts (n = 48) collected over four consecutive seasons by evaluating the metabolomics fingerprinting of the samples using an untargeted approach. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and partial least squares discriminant analysis (PLS-DA) indicated the existence of two key clusters that are linked to the root, stem bark, and leaves. The stem and root chemistry differed from that of the leaves. Seasonal variations were noted in each plant part, with autumn and winter samples closely grouped compared to spring and summer samples in the methanol leaf extracts. Biochemometric analysis could not relate specific compounds to the antimalarial and antitrypanosomal activities of the active extracts, underscoring the intricate interactions among the secondary metabolites. This study further confirms the optimal plant parts to collect in each season for the most effective antimalarial and antitrypanosomal activities.

In Vitro Trypanocidal Effect of the Extract and Isolated Compound of Corymbia torelliana Stem Bark against Trypanosoma brucei brucei (Federe Strain)

African animal trypanosomosis (AAT) is a severe debilitating protozoan disease of livestock and some wild animal species. It is caused by the pathogenic trypanosome species, and transmitted by Tsetse flies. It affects about 60 million cattle in Africa, with about US$1.5 billion in annual losses. Eradication of the disease through chemotherapy and vaccine development has been hindered by parasite antigenic variation and its resistance to drugs. In this study, the crude ethanol stem bark extract of Corymbia torelliana was prepared by cold maceration in 70% Ethanol. It was further purified using the modified Kupchan partition method into n-hexane, Ethyl acetate, N-butanol and Aqueous pure fractions. A pure compound was isolated from the ethyl acetate purified fraction using the column and thin layer chromatographic techniques. The isolated compound was named Makoshi_Mak1. The in vitro antitrypanosomal effects of the crude extract, the purified fractions and the isolated compound were then tested against the blood stream forms of Trypanosoma brucei brucei (Federe strain). 0.2 ml of prepared trypanosome suspension containing about 2x104 parasites was dispensed into each of seven clean test tubes using micropipettes. 0.2ml of prepared Ringer’s solution was also added to stabilize the parasites. Five varying concentrations of the crude extract, the extract fractions and the isolated compound (Makoshi_Mak1) were added to test tubes 1-5. Test tube 6 and 7 served as the negative and positive controls respectively. Aliquots of the contents of each test tube were examined under the microscope at x400 magnification for parasites motility at different time interval for 2 hours. The crude extract was effective at the concentrations of 12 mg/ml, 1.2 mg/ml,0.12 mg/ml and 0.012 mg/ml. the n-hexane fraction had efficacy at 0.12mg/ml and 0.096mg/ml. The Eethyl acetate fractions were effective at 0.12mg/ml, 0.096 mg/ml, 0.048mg/ml and 0.024mg/ml, the N-butanol fraction had efficacy at 0.12mg/ml and 0.096mg/ml, while the Aqueous fraction was effective at 0.2mg/ml, 0.16mg/ml and 0.08mg/ml. Makoshi_Mak1 was effective in vitro against T. brucei beucei at all the tested concentrations of 0.219mg/ml, 0.109mg/ml, 0.055mg/ml and 0.027mg/ml. Efficacy of the extracts and isolated compound was determined by parasite mortality, and was seen to be concentration dependent. Dead parasites appeared to float freely on the fluid environment, while the lysed parasites were seen as floating debris. The results of the study showed that both the crude extract, the purified extract and the pure isolated compound of Corymbia torelliana had in vitro activity against Trypanosoma brucei brucei in a concentration dependent manner. Corymbia torelliana can then be considered as a potential herbal antitrypanosomal drug candidate.

Antibacterial Efficacy of Adamsonia Digitata Stem Bark Crude Extracts Against Staphylococcus Aureus and Shigella Dysenteriae

This research work on the antibacterial efficacy of Adamsonia digitata crude stem bark extracts against Staphylococcus aureus and Shigella dysenteriae, was conducted using Disc diffusion method, on the ethanol and aqueous crude extracts of Adamsonia digitata stem bark against Staphylococcus aureus and Shigella dysenteriae. The crude extracts were effective at all the concentrations (10, 30, 50 and 70 mg/ml) used on the test bacteria. The ethanol crude extract indicates high antibacterial efficacy of 6.0, 12.0, 16,0, 16.0mm and 10.0, 16.0, 16.0 and 16.0mm with all the concentrations used against Staphylococcus aureus and Shigella dysenteriae respectively while the aqueous crude extract also indicates activity of 6.0, 12.0, 16,0, 20.0mm and 12.0, 16.0, 16.0 and 16.0mm all all the concentrations used against Staphylococcus aureus and Shigella dysenteriae. Ciprofloxacin impregnanted antibiotic disc was used as positive control which indicates activity of 18.0 and 20.0 against Staphylococcus aureus and Shigella dysenteriae. The minimum inhibitory concentration (MIC) and the minimum bactericidal study of ethanol and aqueous stem bark crude extracts of Adamsonia digitata against Staphylococcus aureus and Shigella dysenteriae indicates that the plant part used is potent against the test bacteria under study. The phytochhemical screening of the stem bark crude extracts of the plant showed the present of alkanoids, saponins, tannins, flavonoids, terpeniods, glycoside and Anthraquinose. This research work is pointer of the possible used of Adamsonia digitata stem bark crude extracts as remedy for the treatment of infections or disorders cause by these bacteria under study.

Toxicity Studies and in vivo Antiplasmodial Evaluation of Leaf and Stem Bark Extracts of Terminalia avicennioides

Malaria is one of the most serious infectious diseases found in the tropics and subtropics. P. falcifarum resistance to almost all anti-malarial drugs has necessitated the search for anti-malarial compounds. The aim of this study is to investigate the antiplasmodial effects of crude leaf and stem bark extracts of T. avicennioides, a plant utilized by traditional healers in Northern Nigeria to treat malaria and other ailments. Brine shrimp lethality (cytotoxicity) assay and acute toxicity were also evaluated, which are considered a useful tool for preliminary assessment of toxicity in plant extracts. The median lethal dosage (LD50) was calculated by administering different doses of the extract (100–4000 mg/kg) intraperitoneally to three groups of three mice each for 24 hours and observing the animals for physical sign of toxicity. Three models were employed to assess the extracts antiplasmodial effectiveness in vivo against the rodent malaria parasite Plasmodium berghei: suppressive, curative, and repository. The leaf and stem bark extracts were found to show significant toxicity to brine shrimp larvae of Artemia salina in the brine shrimp lethality test with LC50 values of >1000 and 29.6 respectively. Excitation, paw licking, and death were symptoms of acute toxicity of the extracts based on physical and behavioral observations. Death was recorded at 1000 mg/Kg of the stem bark extract and the LD50 is ≤100 mg/Kg body weight while for the leaf extract it was estimated to be ≤600 mg/Kg with no death record. In vivo evaluation revealed that the extracts significantly (P <0.05) reduced parasite count in all models (suppressive, curative and prophylactic), with high average percentage inhibition of parasitaemia (54.68, 58.67 and 65.61%) seen in both T. avicennioides leaf and stem bark extracts, which was comparable to that produced by chloroquine and artesunate, respectively (64.31&93.1; 70.08&80.67; 63.40&75.20%). Both extracts had moderate antiplasmodial activity and were non-toxic to mice and brine shrimps. This finding validates the plant's traditional use in malaria treatment.

Cognitive enhancing effects and anticholinesterase activity of stem bark and leaf extracts of Prunus africana

Alzheimer's disease is ranked among the top five causes of death for old people. Globally, it is approximated that there are 7.7 million new cases of Alzheimer's disease per annum and it is expected that by the year 2050, as many as 1.5% of people will be victims of Alzheimers or other types of dementia. Currently there is no cure for Alzheimer's disease and the conventional therapeutics agents available either have low efficacy or are associated with serious side effects. In the current study, in vivo cognitive advancing and anticholinesterase effects of crude methanol extracts of stem bark and leaf of Prunus africana were investigated in scopolamine treated mice. Passive avoidance task was used to evaluate cognitive enhancing effects of the two plant extracts. Donepezil was used as the standard drug. Scopolamine butylbromide (5 mg/kg bw) was administered intraperitoneally to induce Alzheimer's disease in mice during the study. A completely controlled randomised experimental design was employed in the current study. The two extracts displayed significant anticholinesterase activities and improved cognition in a dose dependent fashion as indicated by escape latency trends. From the current study, it is concluded that methanol extracts of stem bark and leaf of P. africana contain phytochemicals with anticholinesterase activity and cognitive enhancing effects in scopolamine treated mice. The study therefore supports use of leaf and stem bark extracts of P. africana for management of dementia by traditional herbal practitioners.

In vitro uterotonic activities of cold aqueous stem bark extract of Erythrophleum suaveolens on estrogenized uterus of Nulliparous Guinea Pig

Medicinal plants have been used for different purposes. Interests in pharmacological and other scientific screening for the authentication of folkloric claims and drug development has become imperative. This study was aimed at investigating the effect of cold aqueous crude stem-bark extract of Erythrophleum suaveolens (CASBEEs) on estrogenized uterus of nonpregnant Guinea pig. Stem-bark of E. suaveolens were harvested, gently rinsed with clean water and dried under shade in a well-ventilated environment, ground into powder and cold aqueous crude extract of the processed plant material was obtained via maceration. Guinea pigs were estrogenized with 0.2 mg/kg diethylstilbestrol intraperitoneally 24 hours prior to in vitro drug administration. The uterine muscle of the animal was isolated and contractile responses were obtained using different graded volumes with CASBEEs (1 x 10-2 g/ml), oxytocin (1 x 10-4 g/ml), ergometrine (1 x 10-4 g/ml), and Acetylcholine (1 x 10-4 g/ml). The CASBEEs, caused contraction of the isolated estrogenized uterine muscles of the guinea pig in a dose-dependent manner. Maximum contraction (100%) for CASBEEs is at 0.6ml with a final bath concentration of 1.2 x 10-4 g/ml. comparatively, maximum contractile responses for the standard drugs was oxytocin 0.8 (1.6 x 10-6 g/ml) and ergometrine 0.8ml (1.6 x 10-6 g/ml) and Acetylcholine 0.4ml (8 x 10-7 g/ml). Aqueous crude extract of the stem-bark of Erythrophleum suaveolens exerted a dose-dependent and sustained uterotonic effect on estrogenized isolated uterine muscle of nulliparous Guinea pigs, thus authenticating the folkloric claim for its use to induce labour. Further studies on its mechanism of action and its use for fertility control should be explored.

Ex vivo inhibition of PGE2 formation in human blood by four bicyclico [3.2.1] octane neolignans isolated from Aniba firmula bark, two with unusual structural pattern

The phytochemical investigation of the stem bark crude extract of Aniba firmula (Lauraceae) led to the isolation of undescribed bicyclic [3.2.1] octane neolignans, 1 and 2, characterized by unusual bicyclic patterns and two other known bicyclic neolignans 3 and 4. Anti-inflammatory bicyclic [3.2.1] octane neolignans metabolites were previously reported in the literature, and the A. firmula stands out in the Lauraceae family as a source of potentially bioactive compounds. Thus, herein the anti-inflammatory potential of four isolated compounds from A. firmula was accessed via an ex vivo anti-inflammatory model that included plasmatic quantification of the prostaglandin E2 (PGE2) inflammatory mediator. Compounds 2 and 3 exhibited significant anti-inflammatory activity by inhibiting the production of PGE2 in plasma samples, thus by interference with the cyclooxygenase (COX) inflammatory pathway. Therefore, these findings demonstrate that the bicyclic octane neolignan classes [3.2.1] can present anti-inflammatory potential.

Assessment of phytotoxic, genotoxic and enzyme inhibition potential of.

To evaluate Sterculia diversifolia stem bark and leaves for phytotoxic, genotoxic and enzymes inhibition potential. Phytotoxic activity of both stem bark and leaves were screened using Lemna minor. The genotoxic activity of Sterculia diversifolia stem bark and leaves extracts were tested using comet assay protocol while enzyme inhibition activity of crude extract and various fractions of both stem bark and leaves were evaluated using acetyl cholinesterase, lipoxygenase, β-glu-curonidase, urease, xanthine oxidase and carbonic anhydrase. Phytotoxic activity showed significant results in dose dependant manner in both stem bark (ethyl acetate and n-butanol) and leaves (ethyl acetate, n-butanol and n-hexane) fractions. In genotoxic activity, dichloromethane fraction showed significant activity followed by ethyl acetate fraction. Acetyl cholinesterease inhibitory activity showed significant results in both stem bark and leaves fractions, while significant lipoxygenase inhibition was shown by ethyl acetate, dichloromethane, crude extract and n-hexane fractions of both stem bark and leaves. β-glucuronidase, urease and carbonic anhydrase inhibitory activity showed highly significant results in ethyl acetate fraction of both stem bark and leaves, while xanthine oxidase inhibition was shown by dichloromethane fraction of stem bark and leaves extracts. This study emphasizes the important phytotoxic, genotoxic and enzyme inhibition effects of Sterculia diversifolia stem bark and leaves. Hence, it is clear that Sterculia diversifolia stem bark and leaves possess phytotoxic, genotoxic and enzyme inhibitory agents.

Hepatoprotective Effects of Crude Stem Bark Extracts and Solvent Fractions of Cordia africana against Acetaminophen-Induced Liver Injury in Rats.

Cordia africana Lam (Boraginaceae) is widely used in Ethiopian folk medicine for the treatment of different types of liver disorders. Thus, this study aimed to investigate the hepatoprotective effects of an aqueous (CAAE), 80% methanol extracts of C. africana stem bark (CAME), and the solvent fractions of the methanol extract against acetaminophen (APAP)-induced liver injury in rats. Acute toxicity test and APAP-induced lethality test were done on mice of either sex, while APAP dose selection test was done on female rats. Male rats were used for hepatoprotective experiments and the liver injury was induced using 2 g/kg APAP given orally. Serum levels of the liver enzymes and total bilirubin (TB), as well as lipid profiles, were determined. Histopathological examination of the liver tissues was also conducted to confirm the findings of biochemical analysis. Intraperitoneal (i.p.) sodium pentobarbital (SPB)-induced sleeping duration was also used to determine the protective effect of the test substances. Oral administration of APAP resulted in a significant increase in serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), TB, low-density lipoprotein (LDL), total cholesterol (TC), and triglycerides (TGs) and decrease in serum high-density lipoprotein (HDL). Administration of the standard drug, silymarin 100 mg/kg, extracts at doses of 100, 200, and 400 mg/kg and fractions at the dose of 400 mg/kg reversed the serum levels of all parameters to normal. CAME exerted a significant dose-dependent hepatoprotective effect in terms of ALT and AST, while CAAE significant dose-dependent hepatoprotective effect was in terms of AST, ALP, and TGs. The protective effect of the extracts and fractions was also confirmed by histopathological investigations and SPB-induced sleeping time. From the results of the present study, it can be concluded that C. africana stem bark aqueous, 80% methanol crude extracts, and solvent fractions of the methanol extract showed hepatoprotective effects.

Ursane-type triterpenoids, steroids and phenolics from the stem bark and leaves of Nauclea orientalis (L.) L. (Rubiaceae)

Phytochemical investigation of the stem bark and leaf extract of Nauclea orientalis led to the isolation and identification of the three ursane-type triterpenoids rotundic acid, (3β)-3,19,23,24-tetrahydroxyurs-12-en-28-oic acid, 3-oxo-19,23,24-trihydroxyurs-12-en-28-oic acid, the two steroids β-sitosterol, β-sitostenone, the two megastigmane derivatives blumenol A and blumenol A glucoside, the isocoumarine trans-4-hydroxymellein, as well as the flavonole glycoside rutin. From the crude stem bark extract, we performed an anti-bacterial assay against four Gram positive and six Gram negative bacterial strains. An acetylcholinesterase inhibitory assay was performed from the crude extracts and rotundic acid, (3ß)-3,19,23,24-tetrahydroxyurs-12-en-28-oic acid and β-sitostenone. Both assays showed only weak effects. Contact toxicity determined via topical application revealed increased mortality of Spodoptera exigua caterpillars. The chemophenetic significance of ursane-type triterpenoids for the genus Nauclea is discussed briefly.

Effect of Cinnamomum cassia Stem-Bark Extracts on the Synthesis and Secretion of Insulin in RIN-m5F Cells

Cinnamomum cassia (C. cassia) crude stem-bark extracts possess some hypoglycemic properties through inhibiting the uptake of glucose by the intestines and/or enhancing insulin sensitivity. To further investigate the mechanism through which the hypoglycemic effect is exerted, this research explored the influence of the extracts of the crude stem-bark of C. cassia, with various polarities, on particular aspects of glucose consumption as well as glucose-stimulated insulin synthesis and secretion by RIN-m5F cells. All C. cassia extracts did not enhance glucose consumption when compared to the untreated cells (p˂0.001). However, lower concentrations of the extracts significantly increased mRNA levels of Glucose Transporter 2 (GLUT2) and glucokinase genes. Furthermore, all C. cassia extracts significantly stimulated/increased total insulin synthesis (intracellular and secreted) when compared with the control cells, though the effect was not dose-dependent except for water extracts at higher concentration (p<0.001). The results of this investigation revealed that generally, the extracts upregulated the expression of Musculoaponeurotic fibrosarcoma homolog A (MafA), the transcription factor involved in insulin gene expression, while that of the Pancreatic Duodenal Homeobox-1 (PDX-1) gene was mostly suppressed. In concluding, the results of this study proposes that C. cassia stem-bark extracts may exert hypoglycemic effects through stimulating insulin synthesis and secretion in the RIN-m5F pancreatic beta cells.

Effect of castration and testosterone administration on the analgesic efficacies of methanolic extract of Annona Senegalensis and Piroxicam in Wistar rats

Introduction: Analgesic potency of Annona senegalensis (crude stem-bark methanolic extract) and piroxicam on castrated and testosterone-treated male Wistar rats were evaluated. Methods: Twenty-five (25) mature male Wistar rats with average body weight of 184 ± 4.1 g were randomly allocated into five groups of five (n = 5) rats each. Groups I and II were non-castrated (NC) and treated as follows: group I (NC + Annona senegalensis {AS; 910mg/kg}), while group II (NC + piroxicam {PRX; 10mg//kg}). Rats in groups III, IV and V were castrated and treated as follows: castrated (CS) + distilled water {DW; 0.5ml/rat}); (CS + Annona senegalensis {AS; 910mg/kg}); (CS + AS + Testosterone 400µg/rat) respectively. The latent period of pain sensation (analgesic time) was determined using hot plate thermal method, and recorded in seconds. Results: Latent period of analgesia in rats was recorded at 0, 30-, 60-, 90-, and 120-mins post treatment with A. Senegalensis and piroxicam. The serum concentration of testosterone post experiment showed higher levels in group II rats (NC+PRX; {4.3ng/ml}) while group III and V rats had the lowest concentration (CS+DW; CS+AS+TES {2.9ng/ml}) and group IV rats all died after the experiment (CS+AS). The findings of the study indicated that the analgesia effect of A. senegalensis is higher than the piroxicam-induced analgesia (3.8±1.2 vs 1.0±0.0sec, 2.0±1.4 vs 1.0±0.0sec) at 30mins and 60mins in groups I and II respectively. Significance: Castration and testosterone replacement did not affect the Analgesic effect of Annona senegalensis while castration enhanced the latent period of thermal pain sensation.

28 Days Sub Acute Toxicity Studies of the Methanol Stem Bark Extract of Combretum hypopilinum Diels (Combretacea) in Rats

The aim of this study was to evaluate the safety profile of Combretum hypopilinum stem bark extracted with Methanol (70%v/v). Preliminary Phytochemical screening of the crude methanol stem bark extract was carried out, and revealed the presence of secondary metabolites such as steroids, flavonoids and alkaloids. Initial oral acute toxicity test was carried out using the Limit Dose Test to ascertain the safety of the extract in rats. Sub-acute toxicity testing was conducted by 28 days oral administration of 400 mg/kg, 800 mg/kg and 1600 mg/kg body weight to three groups of ten rats. The fourth group was administered distilled water 10 ml/kg. No major changes were observed in body weight of the animals following 28 days of daily oral administration. Biochemical parameters such as Total Protein, Total Bilirubin, Creatinine, Aspartate Transaminase (AST) and Alanine Transaminase (ALT), were found to be within normal ranges. The levels of marker enzymes in the vital organs did not show any significant changes between control and treated groups. Histopathological examination of the major vital organs (liver, brain, and kidney) revealed no significant pathological changes in the treated groups of rats. The results of the present work suggested that the methanol stem bark extract of Combretum hypopilinum is relatively safe for use at the tested doses.

In vitro inhibition of multi-drug resistant Pseudomonas efflux pump by Xylopia aethiopica (Dunal) A. Rich

Global health is under constant threat due to antimicrobial drug resistance. Bacterial Infections caused by Pseudomonas aeruginosa are of importance because of their antibiotics resistance. This study aimed at evaluating the effects of extracts of Xylopia aethiopica (XA) on multidrug-resistant (MDR) Pseudomonas isolates. Fresh samples of XA leaf, stem bark and roots were collected from the botanical garden, University of Ibadan, Nigeria. Dried and pulverized samples were extracted with methanol and partitioned into n-hexane, dichloromethane and ethyl acetate. Phytochemical screening of the extracts was performed by standard methods. Antimicrobial activity and synergistic interaction were determined using microdilution and checkerboard broth dilution methods, respectively. The results revealed that crude methanol extracts of XA leaf, stem bark and root significantly (p&lt;0.05) inhibited the growth of all tested MDR Pseudomonas isolates at 10 mg/mL. At 1 mg/mL, the ethyl acetate fraction of the leaf, and dichloromethane fraction of the roots produced clear zones of inhibition of 12 – 20 mm, and minimum inhibitory concentrations (MICs) of 1 µg/mL and 0.5 mg/mL, respectively. The modulation factor (MF) of ciprofloxacin, dichloromethane fraction of XA roots and ethyl acetate fraction of XA leaf were 4, 8, and 4 on MDR isolates E01006, OAU058 and P. aeruginosa ATCC 27853, respectively. In all tested isolates, but not E01006 and E01024, the fractional MICs of ciprofloxacin/ethylacetate fraction of XA leaf extract combination was not significantly different (p&gt;0.05) compared with ciprofloxacin/verapamil combination. In conclusion, the root and leaf fractions Xylopia aethiopica that demonstrated antimicrobial activity against MDR P. aeruginosa and synergised with ciprofloxacin have the potential to rejuvenate the antimicrobial activity of ciprofloxacin in MDR P. aeruginosa.

Isolation and Characterization of Chemical Constituents from the Crude mExtracts of Crossopteryx Febrifuga (Afzel ex g. don) benth (Rubiaceae) and Evaluation of Their Phytotoxicity and Antioxidant Activity

The aim of this work is to isolate, characterize, and evaluate the antioxidant activity as well as the phytotoxicity of crude extracts and compounds obtained from C. febrifuga. Nine compounds were isolated from leaves, stem bark and roots (methylene chloride/methanol:1/1) crude extracts after repeated silica gel column chromatography corresponding to shanzhiside methyl ester (1), D-mannitol (2), a mixture of two isomeric pentacyclic triterpenes: oleanolic acid (3a) and ursolic acid (3b), 11-methylixoside (4), 3β-urs-12, 20(30)-diene-27,28-dioic acid (5), quinovin glycoside C (6), 3-O-β-D-3-oxo-glucopyranosyl-ursa-12,20(30)-diene-27,28-dioic acid (7) and 3β-(-α-Lrhamnopyranosyloxy)-ursa-12,20(30)-diene-27,28-dioic acid (8). The compound chemical structures were established by studying their 1H and 13C NMR (1D and 2D) and Mass Spectrometry (MS) spectra. The antioxidant capacity of extracts were evaluated using a recently developed electrochemical method consisting in interacting with the superoxide anion radical (O2.-) generated in situ by reduction of air molecular oxygen (O2) in aprotic medium (anhydrous DMF). The antioxidant index is expressed as the amount of extract needed to consume 30% (AI30) of electrogenerated O2.-. The lower is the antioxidant index, the more active the substrate is against O2.-. The stem bark crude extract is the most active with an AI30 of 79.50mg/L while the leaves and root crude extracts showed a moderate activity against O2.- with AI30 of 303.70mg/L and 450.00mg/L respectively. The phytotoxicity study were carried out using a germination test on Lepidium sativum seeds in absence (0mg/L) and in presence of increasing concentrations (10, 100, 500, 1000mg/L) of crude extracts and the isolated compound (1). The following physiological parameters of the seedlings were evaluated 72 hours after the experimentation: germination percentage (G %), germination index (GI), vigor index (VI), relative root growth (RRG %). Root extracts are the most phytotoxic on seed germination with G% ranging from 66.66 to 100% and RRG% from 8.20 to 88.45% while the isolated compound (1) stimulated growth between 10-500mg/L but showed a moderate phytotoxic effect on seedlings above 500mg/L.

Phytochemical and Biological Studies of Bark Extract of Miliusa velutina (Dunal) Hook. f. &amp; Thomson

The main objective of the current research was phytochemical and biological studies of the stem bark of Miliusa velutina (Dunal) Hook. f. &amp; Thomson (Annonaceae). Four purified compounds i.e., friedelin, lupeol, β- sitosterone and caffeic acid were isolated by repeated chromatographic separation and purification of M. velutina. The compounds were identified by analysis of NMR spectral data. The crude dichloromethane extract of stem bark of M. velutina (DEMV) along with its Kupchan partitionates i.e., petroleum ether (PESF), ethyl acetate (EASF), chloroform (CSF) and aqueous (AQSF) soluble fraction were screened for antioxidant, cytotoxic, thrombolytic and antibacterial activities. During the antioxidant activity assay, the AQSF revealed maximum activity with IC50 value of 71.67 μg/ml. The cytotoxicity of plant samples was determined by brine shrimp lethality bioassay, where the maximum cytotoxic activity has been observed for EASF (LC90 = 9.01 μg/ml). In the thrombolytic activity test, the crude dichloromethane extract demonstrated significant efficacy with 46.27% inhibition of clot lysis. In antibacterial screening, the CSF exhibited noticeable inhibitory activity against Shigella boydii with the zone of inhibition 15 mm compared to the standard ciprofloxacin (zone of inhibition = 47 mm).&#x0D; Dhaka Univ. J. Pharm. Sci. 19(2): 125-131, 2020 (December)

GC-MS Analysis and Antimicrobial Spectrum of Stem Bark Extracts of Ficus sycomorus

Aim: This work investigated the gas chromatography-mass spectroscopy of methanolic extract and antimicrobial spectrum of acetone and aqueous crude extracts of Ficus sycomorus stem bark.&#x0D; Place and Duration of the Study: Department chemistry research laboratory and microbiology laboratory, Yobe State University, Damaturu between April 2019 and August, 2019.&#x0D; Methodology: Pure isolates of Staphylococcus aureus, Salmonella typhi, Shigella dysentariae, proteus spp, Pseudomonas auruginosa, Escherichia coli, Bacillus subtilis, Klebsiella pneumonia and a fungi, Candida albicans were collected from National Veterinary Research Institute (NVRI), Vom, Plateau State. The plant was collected and the extract was used in preliminary phytochemical screening, GC-MS analysis and reconstituted for antimicrobial activities testing.&#x0D; Results: The preliminary phytochemical screening revealed presence of saponin, flavonoids, alkaloids, carbohydrates, cardiac glycoside and tannin. GC-MS analysis revealed the presence of heterocyclic members of pyrazole, imidazole, triazole, thiazole and 1, 3-Benzenedicarboxylic acid, bis (2-hethylhexyl) ester. Acetone extract showed the greatest antimicrobial activity against Bacillus subtilis, Pseudomonas aeruginosa, Proteus mirabilis and Shigella spp while the aqueous extract produced antibacterial activity against Bacillus subtilis, Staphylococcus aureus, Klebsiella pneumonia and Candida albicans. Both acetone and aqueous extracts of both stem bark extracts did not exhibit any activity against Escherichia coli. All the extracts were tested at 100mg/ml concentration. The Minimum Inhibitory Concentration (MIC) and the Minimum Bactericidal Concentration (MBC) (125 – 250 mg/ml) of both extracts were higher than those of ciprofloxacin (5.0 mg/ml).&#x0D; Conclusion: There is scientific basis for the uses of F. sycomorus in traditional medicine for the treatment of various diseases as prescribed earlier on and stronger basis for recommendation for its cautious use for the presence of 2-ethylhexyl which could be Carcinogenic. This is first study to identify 2-ethylhexyl from F. sycomorus stem bark extract and recommend cautious use.