Oral toxicity of elaeis guineensis jacq Essay
2.0 LITERATURE REVIEW
Toxicity is an look of being toxicant, bespeaking the province of inauspicious effects led by the interaction between poisons and cells. This interaction may change depending on the chemical belongingss of the poisons and the cell membrane, as it may happen on the cell surface, within the cell organic structure, or in the tissues beneath every bit good as at the extracellular matrix. The toxic effects may take topographic point prior to the binding of the poisons to the critical variety meats such as liver and kidneys. Hence, rating of toxic belongingss of a substance is important when sing for public wellness protection because exposure to chemicals can be risky and consequences to inauspicious effects on human being. In pattern, the rating typically includes ague, chronic, carcinogenic and generative effects ( Asante-Duah, 2002 ) .
As defined by the Organization for Economic Cooperation and Development ( OECD ) , acute toxicity is the happening of inauspicious effects prior to an unwritten disposal of a individual dosage of a substance in a short period clip or of multiple doses given within 24 hours ( Hayes, 2001 ) . The inauspicious effects refer to operational disablement or impairment and biochemical lesions that may interrupt the whole public presentation of an being or restricting the map of an organ to response under critical status. As substance that come ining the being through the unwritten path under a restricted clip and therefore ensuing in inauspicious consequence is known to be orally and acutely toxic, nevertheless the term acute unwritten toxicity is really frequently connected to deadliness and the finding of LD50 ( Walum, 1998 ) .
3.0 MATERIALS AND METHODS
3.1 Methanol infusions readying
3.1.1 Plant stuff sample
Fresh sample of Elaeis guineensis leaves was obtained from Kampung Lekir, Sitiawan, Perak, Malaysia in August 2009 and was authenticated by Mr. Shunmugam A/C Vellosamy from Herbarium Unit, School of Biological Science, Universiti Sains Malaysia. The dried parts of the works including foliages, fruits and flowers were deposited as verifier specimens ( with herbarium figure 11036 ) at the Herbarium Unit, School of Biological Science.
3.1.2 Preparation of the petroleum infusions
The midvein of the Elaeis guineensis foliages were removed before cutting the cusps into pieces. The sample was so washed exhaustively and rinsed with tap H2O and dried in oven at 60 A°C for two to four yearss. The foliages sample was consecutive extracted with methyl alcohol by about adding 100 g of the dried sample ( in all right pulverization signifier ) to 400 milliliter methyl alcohol. The extraction was carried out at room temperature and soaked for four yearss with intermittent stirring during the first twenty-four hours. The infusions were filtered and the procedure of extraction was repeated once more for a 2nd clip by adding another 400 milliliter to the sample residue. The filtrate from each extraction was combined and concentrated under vacuity by rotary evaporator until dark green methyl alcohol infusions produced. The infusions were freeze dried and kept at 4 A°C until usage.
3.2 Acute unwritten toxicity survey of Elaeis guineensis methyl alcohol infusions in mice
The experiment was conducted on 40 healthy Swiss albino mice ( males and females ) weighing 25 to 35 g and aged 8 to 10 hebdomads, acquired from the Animal House, Universiti Sains Malaysia. Those mice were distributed into four groups i.e. both two treated groups and two control groups of opposite sex. The experimental processs associating to the animate beings were authorized by — — – [ Ethical figure: — — – ] before get downing the survey and were conducted under the internationally recognized rules for research lab animate being usage and attention ( EEC Directive of 1986 ; 86/609/EEC ) .
3.2.2 Procedure of acute unwritten toxicity
The mice used in the experiment were selected at random and marked at the dress suits for single designation. Each 10 mice of the same sex were kept in a matte plastic coop, with dimension of 17 A- 27 A- 14 centimeter. All of the coops were located in a room at temperature about 23 A°C with changeless humidness. The room is regulated with rhythms of 12 H of visible radiation and 12 H of darkness. The mice were acclimated to the research lab environment for a hebdomad earlier before get downing the experiment. Drinking H2O and nutrient were provided ad libitum through the experiment except for the short fasting period where the imbibing H2O was still in free entree but no nutrient supply within 12 h prior to intervention. The acute unwritten toxicity of Elaeis guineensis methyl alcohol petroleum infusions was evaluated in mice harmonizing to the process outlined by the Organization for Economic Co-operation and Development ( OECD ) . A individual high dosage of 5000 mg/kg of petroleum infusions was administered to both 10 male mice and ten female mice through unwritten path. The infusions were suspended in a vehicle ( Tween-20 in distilled H2O ) . Following the fasting period, organic structure weight of the mice was determined and the dosage was calculated in mention to the organic structure weight as the volume of the infusions solution given to the mice is 10 mL/kg. Another 10 male mice and ten female mice were allotted with distilled H2O and were regarded as the control groups. Food was provided to the mice about an hr after intervention. The mice were observed in item for any indicants of toxicity consequence within the first six hours after the intervention period, and daily farther for a period of 14 yearss. Surviving animate beings were weighed and ocular observations for mortality, behavioural form, alterations in physical visual aspect, hurt, hurting and marks of unwellness were conducted daily during the period.
3.3 Clinical analysis
3.3.1 Organs and organic structure weight statistical analysis
Completing the 14 yearss period, the whole mice were gently sacrificed. Critical variety meats such as bosom, kidneys, liver, lung and lien, and besides a fragment of the rib coop were isolated and examined for any lesions. All of the single variety meats were weighed and their characteristics were compared between both treated and command groups. Statistical analysis to measure the important difference between both groups was conducted by running a T trial utilizing Microsoft Excel spreadsheet application. The degree of significance used in this analysis is 5 % .
3.3.2 Histopathology of bosom, kidneys, liver, lung, lien, and ribcage
All the critical variety meats and the rib coops isolated from each person were fixed in 10 % buffered formol, routinely processed and embedded in paraffin wax. Paraffin subdivisions ( 5 Aµm ) were cut on glass slides and stained with haematoxylin and eosin. The slides were examined under a light microscope and the exaggerated images of the tissues construction were captured for farther survey.
3.4 Brine runt deadliness trial
3.4.1 Hatching runt
Brine runt eggs, Artemia Salina were hatched in a vas incorporating unfertile unreal saltwater prepared by fade outing 38 g tabular array salt in 1 L distilled H2O. The vas was kept under an inflorescent bulb and facilitated with good aeration for 48 H at room temperature. After hatching, active larvae ( nauplii ) released from the egg shells were collected at the bright side of the vas ( near the light beginning ) by utilizing micropipette. The larvae were isolated from the eggs by aliquoting them in little beaker incorporating the saltwater.
3.4.2 Brine runt trial
The bioactivity of the infusions was monitored by the seawater runt deadliness trial ( Meyer, et al. , 1982 ) to foretell the presence of cytotoxic activity in the compound. The infusions was dissolved in methyl alcohol and diluted with unreal saltwater. The check system was conducted by fixing 10 bijoux bottles filled with 2 milliliters of saltwater each and a double dilution was set up to give a series of concentrations from 100 to 0.195 mg/mL. Potassium bichromate was dissolved in unreal saltwater and functioned as a positive control with concentration runing from 0.1 to 0.9 mg/mL. An aliquot ( 0.1 milliliter ) incorporating approximately 10 to 15 larvae was introduced to each bottle and the apparatus was allowed to go on for 24 h. the bottles were observed, and the dead larvae from each bottles were counted after 6 and 24 h. Based on the per centum of the mortality, the concentration that led 50 % deadliness ( LC50 ) to the larvae was determined by utilizing the graph of average per centum mortality versus the logarithm of concentration ( Latha, et al. , 1998 ) .
3.4.3 Data analysis
The average consequences of mortality per centum of the seawater runt versus the logarithms of concentrations were plotted utilizing the Microsoft Excel spreadsheet application, which besides formulated the arrested development equations. These equations were subsequently used to cipher LC50 values for the samples tested with consideration of value greater than 1.0 mg/mL proposing that the compound is atoxic.
3.5 General sterile techniques
Aseptic techniques involve patterns to minimise the debut of taint when carry oning a laboratorial work. All of the setup and equipments were sterilized before used and environing country is made certain to be clean. Sterilization for the equipments and tools was carried out utilizing autoclave at 120 A°C for 20 proceedingss while the work bench was swept with 70 % intoxicant to forestall taint on working surface.
3.6 Materials and chemicals
Table 3.1 List of stuffs and chemicals used during the survey
3.7 Apparatus and equipments
Table 3.2 List of setup and equipments used during the survey
Distilled H2O machine
4.1 Preparation of methanol infusions of Elaeis guineensis
The Elaeis guineensis foliages were dried and land before extracted with methyl alcohol. Table 4.1 shows the output of the merchandises in weight and the per centum calculated for the ratio of dried and ground stuff per fresh samples and methanol infusions per dried and ground stuff, in respective.
4.2 Lethality and behavioural analysis
The deadliness and toxicity consequence of the methanol infusions of Elaeis guineensis on the mice visual aspect and behavioural form are severally shown in Table 4.2 and table 4.3. There was no decease among the animate being during the observation as besides no important alterations in general visual aspect or behavioural form reported. Furthermore, all the variety meats either of the control or the trial groups are in good form and conditions.
4.3 Organs and organic structure weight statistical analysis
The organic structure weight every bit good as the weights of the critical variety meats of the animate beings were calculated and recorded in Table 4.4. There were no important differences in the alterations of each weight.
4.4 Histopathology analysis of bosom, kidneys, liver, lung, lien, and ribcage
The microscopic constructions of the variety meats depicted through Figure 4.1 to Figure 4.11 show unobtrusive differences between the control and trial group. There were besides no cell debasement or any unfavourable effects observed when viewed under the light microscope utilizing multiple magnification power.
4.5 Brine runt deadliness trial
Brine shrimp deadliness of the methanolic petroleum infusions of Elaeis guineensis are shown in Figure 4.12 and 4.13 and the LC50 values calculated are recorded in Table 4.5. The methanolic petroleum infusions show positive consequence, bespeaking that the samples are biologically active. Crude extracts ensuing in LC50 values of less than 1 mg/mL are considered as significantly active while this suggests that the Elaeis guineensis petroleum infusions have a really low toxicity consequence giving the values of LC50 9.00 and 3.87 mg/mL at 6 and 24 hr severally. Ploting of mortality per centum versus log of concentration for all trials ( Figure 4.12 to 4.14 ) demonstrates an approximative additive correlativity. Furthermore, there is a direct relative relation between the concentration of the infusions and the grade of deadliness. This is shown as the maximal mortalities occurred at a concentration of 100 mg/mL whilst concentration of 0.195 mg/mL merely caused really minor mortalities. As a positive control, K bichromate has proven a important toxicity consequence to the runt as its LC50 reached lower than 1.0 mg/mL. Figure 5.15 depicts the morphology of an Artemia Salina tested by the petroleum infusions, demoing no physical harm occurred to the runt.
Application of Elaeis guineensis leaves as a medical specialty for alternate therapy brings possible to bring around peculiar unwellnesss. Therefore, this survey is successfully performed to guarantee and back up that this works is atoxic and hence is safe to use to human. No acute toxicity exhibited when applied orally at concentration of 5000 mg/kg organic structure weight. This has been demonstrated by the normal behaviour and histological observations in reaction to the petroleum infusion when applied to the animate beings. Another trial to foretell the toxic potency of this works is the seawater runt deadliness trial. The consequence based on this trial besides supports that Elaeis guineensis infusion is atoxic and moreover, it reveals that this works has responded actively in bioassay activity. These consequences are of import as they could function as a footing for farther surveies related to pharmacological research of Elaeis guineensis.
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