Chemical profiling of bioactive compounds in the methanolic extract of wild leaf and callus of Vitex negundo using gas chromatographymass spectrometry

BACKGROUND The investigation of plant-based therapeutic agents in medicinal plants has revealed their presence in the extracts and provides the vision to formulate novel techniques for drug therapy.Vitex negundo(V.negundo),a perennial herb belonging to the Varbanaceae family,is extensively used in conventional medication.AIM To determine the existence of therapeutic components in leaf and callus extracts from wild V.negundo plants using gas chromatography-mass spectrometry(GCMS).METHODS In this study,we conducted GC-MS on wild plant leaf extracts and correlated the presence of constituents with those in callus extracts.Various growth regulators such as 6-benzylaminopurine(BAP),2,4-dichlorophenoxyacetic acid(2,4-D),α-naphthylacetic acid(NAA),and di-phenylurea(DPU)were added to plant leaves and in-vitro callus and grown on MS medium.RESULTS The results clearly indicated that the addition of BAP(2.0 mg/L),2,4-D(0.2 mg/mL),DPU(2.0 mg/L)and 2,4-D(0.2 mg/mL)in MS medium resulted in rapid callus development.The plant profile of Vitex extracts by GC-MS analysis showed that 24,10,and 14 bioactive constituents were detected in the methanolic extract of leaf,green callus and the methanolic extract of white loose callus,respectively.CONCLUSION Octadecadienoic acid,hexadecanoic acid and methyl ester were the major constituents in the leaf and callus methanolic extract.Octadecadienoic acid was the most common constituent in all samples.The maximum concentration of octadecadienoic acid in leaves,green callus and white loose callus was 21.93%,47.79%and 40.38%,respectively.These findings demonstrate that the concentration of octadecadienoic acid doubles in-vitro compared to in-vivo.In addition to octadecadienoic acid;butyric acid,benzene,1-methoxy-4-(1-propenyl),dospan,tridecanedialdehyde,methylcyclohexenylbutanol,chlorpyrifos,n-secondary terpene diester,anflunine and other important active compounds were also detected.All these components were only available in callus formed in-vitro.This study showed that the callus contained additional botanical characteristics compared with wild plants.Due to the presence of numerous bioactive compounds,the medical use of Vitex for various diseases has been accepted and the plant is considered an important source of therapeutics for research and development.

Extraction of valuable metals from low-grade nickeliferous laterite ore by reduction roasting-ammonia leaching method

Nickel and cobalt were extracted from low-grade nickeliferous laterite ore using a reduction roasting-ammonia leaching method.The reduction roasting-ammonia leaching experimental tests were chiefly introduced,by which fine coal was used as a reductant.The results show that the optimum process conditions are confirmed as follows:in reduction roasting process,the mass fraction of reductant in the ore is 10%,roasting time is 120 min,roasting temperature is 1 023-1 073 K;in ammonia leaching process,the liquid-to-solid ratio is 4:1(mL/g),leaching temperature is 313 K,leaching time is 120 min,and concentration ratio of NH3 to CO2 is 90 g/L:60 g/L.Under the optimum conditions,leaching efficiencies of nickel and cobalt are 86.25% and 60.84%,respectively.Therefore,nickel and cobalt can be effectively reclaimed,and the leaching agent can be also recycled at room temperature and normal pressure.

Optimization of extraction method for quantitative analysis of Si/Al in soil phytoliths

Elemental information carried by phytoliths plays a crucial indicative role in geochemical research.For instance,it serves as an indicator of the carbon pool effect in phytoliths and aids in the elucidation of silicon sources.However,early extraction methods for soil phytoliths primarily focused on obtaining their morphological and quantitative information,lacking efficient techniques for quantitative elemental analysis.In this study,we aimed to extract Si/Al information from soil phytoliths.Considering the need for complete extraction of phytolith,six extraction methods were developed and further by alkaline dissolution to determine Si/Al.Six methods were compared in terms of enrichment capacity,the weight of extracted phytoliths,and Si/Al differences.The results indicated that the addition of Ammonia-Catechol in the commonly used heavy liquid flotation method effectively improved phytolith extraction capability and the accuracy of Si/Al results.Additionally,the inclusion of an acetic acid step before alkaline dissolution further removed surface-adsorbed impurities and enhanced the analytical quality of Si/Al in phytolith.The comparison of the data in this study with other published data shows that our method is relatively robust.The improved method proposed in this study can provide a new idea for the quantitative analysis of other elements in soil phytoliths.

Effects of Methanolic and Aqueous Extracts of Griffonia simplicifolia (Fabaceae) on the Inhibition of Falciformation of Human Hb SS Erythrocytes

Sickle cell disease (SCD) is a genetic blood disorder that affects the shape and transportation of red blood cells (RBCs) in blood vessels, leading to various clinical complications. Sickle cell disease is a widespread genetic disease in Black Africa. The objective of this study was to evaluate the sickling inhibition activity of Griffonia simplicifolia. Quantitative and qualitative tests were used to determine the major groups of secondary metabolites present in the Griffonia simplicifolia leaves extracts and the modified Emmel test was used to perform the study of sickling inhibition activity. The OECD 423 toxicity study showed that at a single dose of 2000 mg/kg bw, Griffonia simplicifolia leaves extracts are not toxic. All tested substances inhibit erythrocyte falciformation in a dose-dependent manner. The percentages of inhibition were 50.35%, 73% and 94.23% for aqueous extract, hydromethanolic extract and phenylalanine respectively at the concentration of 15 mg/mL. The methanolic extract (70%) had higher activity compared to the aqueous extract. These results suggest that the Griffonia simplicifolia extracts have some potential to be used as alternative antisickling therapy in SCD management.

Process and kinetics of the selective extraction of cobalt from high-silicon low-grade cobalt ores using ammonia leaching

An ammonia-based system was used to selectively leach cobalt(Co)from an African high-silicon low-grade Co ore,and the other elemental impurities were inhibited from leaching in this process.This process was simple and environmentally friendly.The results revealed that the leaching ratio of Co can reach up to 95.61%using(NH_(4))_(2)SO_(4)as a leaching agent under the following materials and conditions:(NH_(4))_(2)SO_(4)concentration 300 g/L,reductant dosage 0.7 g,leaching temperature 353 K,reaction time 4 h,and liquid-solid ratio 6 mL/g.The leaching kinetics of Co showed that the apparent activation energy of Co leaching was 76.07 kJ/mol(i.e.,in the range of 40-300 kJ/mol).This indicated that the leaching of Co from the Co ore was controlled by an interfacial chemical reaction,and then the developed leaching kinetics model of the Co can be expressed as 1-(1-α)^(1/3)=28.01×10~3×r_0^(-1)×C_((NH_(4))_(2)SO_(4))^(1.5)×exp(-76073/8.314 T)×t,whereαis the leaching ratio(%)of Co,r_0 is the average radius(m)of the Co ore particles,T is the temperature(K),and C_((NH_(4))_(2)SO_(4))is the initial reactant concentration(kg/m^(3)).

Solvent Extraction of Nickel and Cobalt from Ammonia-Ammonium Carbonate Solution by Using LIX 84-ICNS

In the present paper, separation of nickel and cobalt in ammonia-ammonium carbonate solution that simulates pregnant leach solution of Caron Process by solvent extraction using LIX 84-ICNS was studied. LIX 84-ICNS is a novel extractant which is still being studied, especially for nickel and cobalt separation in ammonia-ammonium carbonate solution. A series of solvent extraction tests were performed at various equilibrium pH, temperature, extractant concentration, and volume ratio of organic to aqueous solution (O/A ratio). The investigation results show that the highest nickel and cobalt extraction percentages of 99.8% and 90.3% were obtained from the extraction test at equilibrium pH of 8.75, temperature of 55°C, extractant concentration of 40% (v/v) and O/A ratio of 1/1, respectively. Oxidation of cobalt in aqueous solution prior to extraction is needed to minimize co-extraction of cobalt. Co-extracted cobalt can be decreased from 90.3% to 30.3% by mixing 1% (v/v) H2O22 in aqueous solution prior to the extraction stage. It was found that nickel and cobalt extractions by LIX 84-ICNS are endothermic processes with enthalpy changes of +171.03 and +7.64 kJ/mole, respectively. Based on constructed McCabe-Thiele Diagram, nickel extraction level of more than 99.9% can be obtained in 2 stages at O/A ratio of 0.5. The highest stripping percentages of nickel and cobalt of 98.82% and 3.16% respectively were obtained at 200 g/l H2SO4 stripping agent.

Study on Pu-erh Tea Methanol Extracts on Antibacterial Activity of Several Plant Pathogens

[Objective] The aim was to study Pu-erh tea methanol extracts on antibacterial activity of several plant pathogens and provide reference for application of Pu-erh tea in agriculture disease prevention and control.[Method]The inhibitive effect of Pu-erh tea methanol extracts on several plant pathogens was studied by adopting mycelium growth rate method.[Result] The results showed that Pu-erh tea methanol extracts had inhibitive effects on Thanatephorus cucomeris(Frank)Donk,Fusarium oxysporum f.sp.niveum,Fusa...

Anti-diabetic and anti-cholesterolemic activity of methanol extracts of three species of Amaranthus

Objective:To investigate the anti-diabetic and anti-cholesterolemic activity of methanol extracts of leaves of Amaranthus caudatus,Amaranthus spinosus and Amaranthus viridis in normal and streptozotocin(STZ) induced diabetic rats.Methods:In this study,the anti-diabetic and anti-cholesterolemic activity of methanol extracts of leaves of all three plants was evaluated by using normal and STZ induced diabetic rots at a dose of 200 mg/kg and 400 mg/kg p.o.daily for 21 days.Blood glucose levels and body weight were monitored at specific intervals,and different biochemical parameters,serum cholesterol,serum triglyceride,high density lipoprotein, low density lipoprotein and very low density lipoprotein were also assessed in the experimental animals.Histology of pancreas was performed.Results:It was found that all the three plants at 400 mg/kg dose showed significant anti-diabetic and anti-cholesterolemic activity(P<0.01),while at 200 mg/kg dose less significant anti-diabetic activity(P<0.05) was observed.Concluslons: Methanol extracts of Amaranthus caudatus,Amaranlhus spinosus and Amaranthus viridis showed significant anti-diabetic and anti-cholesterolemic activity,which provides the scientific proof for their traditional claims.

Antidiabetic activity of methanolic bark extract of Alhizia odoratissima Benth.in alloxan induced diabetic albino mice

Objective:To evaluate the antidiabetic potential of methanolic extract of Albizia odoratissima Benth.bark in alloxan induced diabetic mice.Methods:Group-Ⅰ(normal control) mice received only basal diet without any treatment.In Group-Ⅱ(Diabetic control) mice,diabetes was induced by alloxan(150 mg/kg i.p.) and received only Tween 80.5%v/v in normal saline. Group-Ⅲand Group-Ⅳmice received metformin(10 mg/kg) and gliclazide(10 mg/kg) as standard drugs.Group-ⅤandⅥmice received methanolic bark extract of Albizia odoratissima at doses of 250 and 500 mg/kg body weight p.o.,respectively.Results:The results of the study indicates that Albizia odoratissima bark extract significantly(P【0.01) reduced the blood sugar level.The bark extract also significantly reduced the levels of serum cholesterol,triglycerides, serum glutamic-oxaloacetic transaminase,serum glutamic-pyruvic transaminase,alkaline phosphatase and decreases level of total proteins in alloxan induced diabetic mice.Conclusions: Methanolic extract of Albizia odoratissima has protective effects on the protection of vital tissues(pancreas,kidney,liver,heart and spleen),thereby reducing the causation of diabetes in experimental animals.

Seasonal variation for the antidiabetic activity of Loranthus micranthus methanol extract

Objective:To determine the season in which the Eastern Nigeria mistletoe,Loranthus micranthus,parasitic on Persea americana possesses optimum antidiabetic activity and to determine the seasonal variation in the constituents.Methods:The antidiabetic activities of the aqueous methanol extracts of the leaves of Eastern Nigeria mistletoe,Loranthus micranthus, harvested in two seasons of the year,the onset of rainy season(April) and the peak of rainy season(July) were compared.The tests were carried out on six(6) groups(A-F) of alloxaninduced diabetic rats.Groups A and B received 200 mg/kg and 400 mg/kg of the April sample extracts respectively while groups C and D received same doses of the July sample extracts. Group E and F which were the positive and negative controls received 10 mg/kg of glibenclamide and 2 ml/kg of 3%tween 20 respectively.The blood glucose levels of the animals were monitored hourly with a glucometer for six hours.The phytochemical analysis of the plant extracts were also carried out by standard procedures.Results:The results showed that group A and B exhibited significant(P【0.05) percentage reduction in the fasting blood sugar(FBS) level of the animals(38.9%and 39.2%respectively) with maximum reduction observed at the 5th and 6th hour respectively compared to glibenclamide(71.3%).Group C showed no significant (P】0.05) FBS reduction(15.9%) while group D exhibited highly significant(P【0.01) reduction (47.5%) with the maximum reduction occurring after 6 hours.The phytochemical analysis of the crude methanol extracts revealed the presence of carbohydrates,glycosides,saponins, tannins,flavonoids,steroids,terpenoids,acidic compounds,resins and oils.These were present in different proportions in both seasons.Conclusions:This study shows that there is a seasonal, dose-dependent variation in the chemical composition viz-a-viz the antidiabetic activity of the plant under study.This activity is highest at the peak of the rainy season.

Extractive Distillation of Methyl Acetate-Methanol Azeotrope Using [DMIM]DMP as Solvent

In order to separate methyl acetate and methanol azeotrope, the ionic liquid(IL) 1,3-dimethylimidazolium dimethylphosphate([DMIM]DMP) was used as the solvent. The Aspen Plus software was used to design and optimize the extractive distillation process. Under the optimized conditions, the mass fractions of methyl acetate and methanol were both above 99.5%. Compared with the dimethyl sulfoxide(DMSO) process, the [DMIM]DMP process has the advantages of saving energy and lower equipment investment cost. The result shows that using [DMIM]DMP as the solvent to separate a methyl acetate and methanol azeotrope has better prospect in industrial application.

In vitro evaluation of antimicrobial and antioxidant activities of methanolic extract of Jasminum humile Leaves

Objective:To evaluate in vitro antimicrobial and antioxidant activities of methanolic extract of Jasminum humile（J.humile） leaves extract.Method:Methanolic extract of J.humile was evaluated for its antimicrobial activity by using agar well diffusion method & their possible antioxidant assay by two complementary test systems,namely DPPH and hydrogen peroxide scavenging activity.These various antioxidant activities were compared to standard antioxidants such as ascorbic acid for both the tests.Results:In the DPPH & hydrogen peroxide scavenging activity,the IC50 value of methanol extract was 70.43μg/mL & 60.79μg/mL respectively. Further,the extract showed inhibitory activity for Gram-positive and negative bacteria at different concentrations.The maximum antibacterial activity of extract was exhibited against Staphylococcus aureus（S.aureus） at concentration 50 mg/mL when compared with ciprofloxacin Conclusions:These results clearly indicate that J.humile is effective in scavenging free radicals and has the potential to be a powerful antioxidant.Thus,the results obtained in the present study indicate that J.humile leaves extract could be considered as a potential source of natural antioxidants and that could be used as an effective source against bacterial diseases.

Anticancer activity of Mahonia leschenaultii methanolic root extract and berberine on Dalton’s ascitic lymphoma in mice

Objective: To evaluate the effects of methanol root extract of Mahonia leschenaultii and berberine of Mahonia leschenaultii on Dalton’s ascitic lymphoma in Swiss Albino mice. Methods: The methanol root extracts of Mahonia leschenaultii(200 and 400 mg/kg) were given orally, and berberines(10 and 20 mg/kg) were injected intra-peritoneally for 14 successive days in tumor bearing mice. Hematological parameters(white and red blood cells, haemoglobin level, granulocytes, and agranulocytes), lipid parameters(total cholesterol and triglycerides), serum enzymes(serum glutamate pyruvate transaminases, serum glutamate oxaloacetate transaminases, and alkaline phosphatise) and mean survival and solid tumor growth were determined and compared with untreated mice. 5-fluorouracil(20 mg/kg) was used as a reference standard drug. Results: Mahonia leschenaultii and berberine reduced the hematocrit significantly. Furthermore, Mahonia leschenaultii and berberine improved the survival of mice significantly and restored the affected hematological and lipid parameters similar to the normal levels. Conclusions: These observations show a strong anticancer effect of methanol root extract of Mahonia leschenaultii and berberine in suppressing Dalton’s ascitic lymphoma cancer cell growth in a mouse model by controlling haematological, lipid, serum enzymes, and other derived parameters effectively.

Antibacterial activity of plant methanolic extracts on a field isolate of <i>Pseudomonas syringae</i>pvtomato from the Casablanca region (Morocco)

A bacterial field isolate recovered from infected tomato plants in a green-house at Sidi Rehal, a region near Casablanca city (Morocco), was identified as the gammaproteobacterium Pseudomonas syringae pv. tomato DC3000 strain, the causal agent of bacterial speck. The bacterial isolate was characterized by morphological, biochemical and molecular biological tests, its growth curves carried out in various culture media, and its phytopathogenicity verified by infection tests. A screening was performed to evaluate the antibacterial activity of methanolic extracts of 12 selected Moroccan plants against the P. syringae pv. tomato DC3000 isolate, and Agar-well diffusion and Broth microdilution methods were used to determine minimum inhibitory and minimum bactericidal concentrations. Among the methanolic extracts tested, only those of Nigella sativa, Geranuim robertianum, Aizoon canariense and Rubia peregrine showed clear inhibitory and bactericidal activities, although the highest values were achieved with N. sativa, a plant used in Morocco as a spice, condiment and medicinal treatment.

Analgesic Activities of Methanol Extract of <i>Terminalia chebula</i>Fruit

Pain is a sensation which in many cases represents the only symptom for the diagnosis of several disorders. Throughout ages man has used many different remedies for pain relief, among which medicinal herbs are most common due to their efficacy and relative safety due to their natural origin. In present study methanol extract of Terminalia chebula fruit was evaluated for its analgesic effects in albino mice. The extract was prepared using whole dried powdered fruit with seeds in absolute methanol. It was then subjected to rotary evaporator under reduced pressure to get concentrated extract;different doses (300, 500 and 1000 mg/kg) were assessed for analgesic activity by tail immersion technique and acetic acid induced writhing test. Similar doses of Aspirin were used as standard drug (positive control) for comparison. Results of both tail flick method and acetic acid induced writhing test revealed that T. chebula fruit extract possessed varying degree of analgesic activity significant at 300 mg/kg and highly significant at 500 and 1000 mg/kg in comparison to control. The results were almost similar to standard drug. In acetic acid induced writhing test, maximum inhibition of writhing was observed at 1000 mg/kg where the number of writhes decreased from 14.1 to 5.2 indicating 63.1% inhibition. Above findings suggest that T. chebula fruit extract possesses significant analgesic activity in albino mice with reference to positive and negative control groups;however further studies on a large number of animals with clinical trials are required to confirm safe and effective use of this fruit extract in humans for analgesic purpose.

Evaluation of antidepressant activity of methanolic and hydroalcoholic extracts of Acorus calamus L.rhizome through tail suspension test and forced swimming test of mice

Objective:Acorus calamus(AC)L.(Araceae)is an annual semi-aquatic and aromatic plant found in Europe,North America and Asia.Its rhizomes are often used by Native Americans,Americans,and Chinese as well as by other cultures.Ethnobotanical studies and documents have shown their use in various disease treatments,such as insomnia,mental disorders,diabetes mellitus,epilepsy,inflammation,asthma,neuropathic pain,and diarrhea.In this study,the antidepressant activity of methanolic and hydroalcoholic extracts of the AC rhizome part in mice was investigated.Methods:Three doses of methanolic extract of AC rhizome(MEACR)(25,50 and 100 mg/kg b.wt),three doses of hydroalcoholic extract of AC rhizome(HAACR)(100,200 and 400 mg/kg b.wt),and standards(imipramine,15 mg/kg b.wt and fluoxetine,20 mg/kg b.wt)was daily oral administration to the mice for consecutive 14 days.The extract effect on the immobility time was monitored by a tail suspension test(TST)and a forced swimming test(FST).Monoamine oxidase(MAO)levels were also analyzed using standard methods.Results:The optimum antidepressant activity was viewed at 100 mg/kg b.wt of MEACR extract and400 mg/kg b.wt of HAACR extract with 23.82%and 20.59%immobility period reduction,respectively.Besides,the extracts weakened the FST-induced elevation of MAO activity significantly and returned to near-normal levels of neurotransmitters in the brain.100 mg/kg b.wt or above of MEACR extract significantly prevented the MAO-A and MAO-B activities in mice brain at a dose-dependent fashion.But,just 400 mg/kg b.wt of HAACR extract prevented the activity of MAO-A and MAO-B.Fluoxetine and imipramine showed a tendency to prevent the activity of MAO-A and MAO-B.Conclusion:This study suggests that AC rhizome extract mediated antidepressant activity by modulating the central neurochemical and hypothalamic-pituitary-adrenal(HPA)axis in response to FST and TSTinduced stress.Therefore,AC rhizome extract can be used as a valuable plant supplement to treat depressive disorders.

Characterization and performance of Cu/ZnO/Al_2O_3 catalysts prepared via decomposition of M(Cu,Zn)-ammonia complexes under sub-atmospheric pressure for methanol synthesis from H_2 and CO_2

Methanol synthesis from hydrogenation of CO2 is investigated over Cu/ZnO/Al2O3 catalysts prepared by decomposition of M（Cu,Zn）-ammonia complexes （DMAC） at various temperatures.The catalysts were characterized in detail,including X-ray diffraction,N2 adsorption-desorption,N2O chemisorption,temperature-programmed reduction and evolved gas analyses.The influences of DMAC temperature,reaction temperature and specific Cu surface area on catalytic performance are investigated.It is considered that the aurichalcite phase in the precursor plays a key role in improving the physiochemical properties and activities of the final catalysts.The catalyst from rich-aurichalcite precursor exhibits large specific Cu surface area and high space time yield of methanol （212 g/（Lcat·h）;T=513 K,p=3MPa,SV=12000 h-1）.

Assessments of Immunological Activity of Achillea Millefolium Methanolic Extract on Albino Male Mice

Achillea millefolium （Asteraceae） is a permanent herb highly recognized in traditional medicine for its anti-oxidant and anti-inflammation properties. However, studies on phytochemical constituents of A. millefolium underlying these properties are scarce. The present work focuses on examining the effect of methanol extract of A. millefolium L. on total and differential blood cells account on albino male mice. The results showed the methanol extract increased the account of lymphocyte, and monocyte cells, and total account as well as this extract showed high decrease in the oxidative stress of MTX after the interfere between the extract and MTX due to increase in the leucocyte cells compared with controls. Concluded from these results that methanol extract of A. millefolium has ability enhancement in leucocyte cells in the blood and it has detoxification effect of MTX.

Larvicidal activity of essential oil and methanol extract of Nepeta menthoides against malaria vector Anopheles stephensi

Objective:To investigate the larvicidai activity of essential oil and methanol extract of the Nepeta menthoides（N.menthoides） against main malaria vector,Anopheles stephensi（An. stephensi）.Methods:The essential oil of planl was obtained by Clevenger type apparatus and the methanol extract was supplied with Percolation method,larvicidai activity was tested by WHO method.Twenty five fourth-instar larvae of An.stephensi were used in the larvicidai assay and four replicates were tested for each concentration.Five different concentrations of the oil and extract were tested for calculation of LC50 and LC50 values.Results:The LC50 and LC90 values were determined by probit analysis.IC50 was 69.5 and 234.3 ppm and LC50 was 175.5 and 419.9 ppm for the extract and essential oil respectively.Conclusions:According to the results of this study methanolic extract of plant exhibited more larvicidai activity than essential oil.This could be useful for investigation of new natural larvicidai compounds.

The effects of methanol extract of cleistopholis patens on the reproductive system of female Wistar rats

Objective:In Port Harcourt and its environs,extracts of Cleistopholis patens are used by traditional medicine healers for the treatment of menstrual irregularities and other gynaecological disorders.The objective of this study was therefore to determine the effects of orally administered methanol extract of the stem bark of Cleistopholis patens on the reproductive organs of non-pregnant albino(Wistar) rats.Methods:3g/kg(low dose) and 6g/kg(high dose) of the extract were administered orally,daily to two different groups of animals,respectively, over a period of 28 days.A third(control) group of animals received distilled water only,orally over the same period.Five animals from each of the groups were sacrificed on day 8,15 and 29.Venous blood samples and reproductive organs respectively were taken from each group of sacrificed animals for hormonal and histopathological analysis.Results:Results of the hormonal assay revealed a general increase in the levels of Follicle Stimulating Hormone(FSH),Luteinizing Hormone(LH),progesterone,and estrogen.The highest levels were noticed in the animals sacrificed on the 29th day(LH =5.48±0.04 IU/L;FSH =3.80±0.00 IU/ L;Progesterone =7.14±0.15 nmol/L;Estrogen = 0.168±0.002 nmol/L).These increases were statistically significant compared to those of the control animals(LH =2.90±0.00 IU /L;FSH = 1.28±0.02 IU/L; Progesterone = 3.80±0.00 nmol/L;Estrogen = 0.130±0.002 nmol/L;P【0.05),and were also dose dependent. Results of the histopathological studies showed presence of chronic inflammatory cells in the tissues of the fallopian tubes and uterus on the 29th day.However,no changes were observed in the ovaries.Conclusion: The administration of the extract produced a dose and time-dependent increase in FSH,LH,progesterone and estrogen levels.We postulate that these observed effects may have been induced by the phytoestrogens (known to have 1/1 000 th of the efficacy of natural oestrogens) in the extract.The hormonal and histopathological changes may explain the effects described by patients following ingestion of extracts of this plant in traditional medical practice.However,it remains to be determined if these effects are harmful or beneficial in disease conditions.