PHARMACOGNOSTIC AND PHYTOCHEMICAL EVALUATION AND ANTIPYRETIC ACTIVITY OF LEAVES OF LANTANA CAMARA LINN .

Lantana camara is a heavily branched shrub that can grow in compact clumps, dense thickets or as a climbing vine. The plant is Antidiabetic, Antiseptic, Anti-inflammatory, Antipyretic, Carminative, Anti-spasmodic, Diphoretic and are useful for cuts, wounds, ulcers, swelling, tumours and rheumatism. Evaluation of fresh, powdered and anatomical sections of the leaves was carried out to determine the morphological, microscopical and phytochemical profiles. Leaves of L . camara Linn. are dark green with characteristic odour, opposite, blade ovate, 4-10cm long, with coarse surfaces and toothed margins etc. Microscopy shows covering trichomes, Sclerenchyma, collenchyma, xylem, phloem etc. Phytochemical evaluation revealed the presence of alkaloids, triterpenoid, essential oil, flavanoids, carbohydrates, tannins and saponins. The investigations also included evaluation of physical parameter such as the moisture content, ash values, and extractive value. Concerning on the Antipyretic activity, Lantana camara ethanolic and ethyl acetate extract start lowering the body temperature from 1.5 th hour, while between 2 nd and 3 rd hour both the extract showed significant (P<0.01) Antipyretic activity when compared with the negative control group by one way ANOVA followed by dunnett’s t-test.


Introduction
Any plant and its parts containing any substance which can be used therapeutically, or can be used as raw material for Chemical/Pharmaceutical synthesis is categorized as herbal medicine. 1,2. Lantana camara L. (Verbanaceae), universally known as wild or red sage is the most widespread species of this genus and regarded both as a notorious weed and a popular ornamental garden plant. However, it is listed as one of the important medicinal plants of the world. L. camara whole plant and plant parts viz.,leaves, flowers, and essential oils have been thoroughly studied for their chemical compositions. 3,4,5,6,16,17. The methanolic extract of Lantana camara leaves shown healing of gastric ulcers and also prevents development of duodenal ulcers in rats. Extracts of the fresh leaves are antibacterial and are traditionally used in Brazil as an antipyretic, carminative and in the treatment of respiratory system infection . Anti Diabetic, Anti spasmodic, Diaphoretic, Carminative, Tonic and useful in the treatment of titanus, vitiated of fresh roots is a good gargle for odontalgia and it is used by hill tribes for all types of dysentery. Powdered leaves are useful for cuts wounds ulcers and swelling and infusion of the leaves is good for bilious fever, eczema and eruption. the fruits are useful in fistula, pastules, tumours and reuhmatism. 18 A small quantity of powder was kept on a slide and after mounting on glycerine, 10min were provided as spared out time. Finally, it was observed for powder microscopical characters. 12

Physiochemical studies
Physicochemical studies include ash value and extractive value to determine the quality and purity of the powder of plant of Lantana camara Linn.

Determination of Ash-value 2.3.1.1. Total ash:
Weigh accurately previously weighed and tarred crucible. Add 2-4gm of ground material. Ignite the material by increasing the heat to 500-600 o C. Cool in desiccators and weigh. If carbon free ash cannot be obtained in this manner, cool the crucible and moisten the residue with 2ml. of water or a saturated solution of ammonium nitrate R. Dry on a water bath or hot plate and ignite. Allow the residue to cool for 30min. and weigh. Calculate content of total. 10 2.3.1.2. Acid-insoluble ash: To the crucible containing total ash, add 25 ml of hydrochloric acid, cover with a watch glass and boil gently for 5 minutes. Rinse the watch glass with 5ml of hot water and add this liquid to crucible. Collect the insoluble matter on an ash less filter paper and wash with hot water until the filtrate is neutral. Transfer the filter paper containing the insoluble matter to the original crucible, and then ignite at 450-500 0 C to constant weight. Cool

Determination of solvent Extractive value:
Weigh accurately, about 4g of coarsely powdered air-dried material to a glass stoppered conical flask. Macerate with 100ml of the required solvent for the given plant material for 6 hour, shaking frequently then allow to stand for 18 hour. Filter rapidly taking care not to lose any solvent, transfer 25 ml of the filtrate to a treated fiat-bottomed dish and evaporate to dryness an a water bath. Dry at 10 o Celsius for 6 hour, cool in a desiccators for 30 minutes and weigh without delay. Calculate the contents of extractable matter in mg per g of air-dried material. 15

Extraction of plant leaf material:
The powdered plant leaf material was subjected to successive solvent extraction taking from polar to nonpolar solvents like water, ethanol, methanol, acetone, chloroform, benzene and petroleum ether. 20gms of powdered plant material was subjected to soxhlet extraction for 8 hrs with 250ml of the various solvents. The extracts obtained were later kept for evaporation to remove the excessive solvents. These extracts were stored in a cool dry place for the analysis for the presence of preliminary pytochemicals. 15

Phytochemical screening
The aqueous and alcoholic extract of the powdered drug were subjected to various qualitative tests for the identification of various plants constituents present in that species.

Thin layer chromatography:
TLC is most widely used technique in biological, chemical and pharmaceutical laboratories.In TLC methods, the separation of the components in the material is based principally on adsorption, or partition, or combination of both, depending on the adsorbent and the solvent used. TLC is prepared on Glass plates of good quality, slurry is prepared which is a mixture of silicagel G(stationary phase) and water and spread on the plates, normally a thickness of 0.25mm is used for analytical purpose, after setting plates are activated by keeping in an oven at 100 degree for 1 hour. After activation spot of the sample is placed above 2cm on the base of the plate, and the plate is kept in the development tank contaning mobile phase of different solvents. After the development of TLC plates the spots are visualized. for detecting,colourless spots are observed by keeping the plates in Iodine chamber. 7,8 The ethanolic extract is referred to produce TLC because these extract only having more active constituents. The ethanolic extract showed resolution of spot with following solvent system. 7 Ethyl acetate: Methanol= (80:20) 2.7. Antipyretic activity 2.7.1. Animal care and handling: The experiment was carried out on albino rabbits of 5-8 months, of both sexes, weighing between 1.5 to 1.6 kg. They were provided from Sapience Bioanalytical Research Lab, Bhopal, (M.P.) and were kept in mesh bottom iron cages to avoid caprophagy. The animals were acclimatized to the standard laboratory conditions in cross ventilated animal house at temperature 25±2°C relative humidity 44 -56% and light and dark cycles of 11 and 13 hours respectively fed with cauliflower, cabbage, carrot and tap water for 10 days before the experiment. Food and water were withdrawn 14 hours prior to the experiment. The experiment was approved by the institutional ethics committee and as per CPCSEA guidelines (approval no. 1413/a/11/CPCSEA).

2.7.2.
Grouping and treatments of experimental animals: Four animals in each group were taken and total four groups were grouped. Group I (-ve control, vehicle treated), Group II (standard, paracetamol 100mg/kg treated), Group III (ethanolic extract 300mg/kg treated), Group IV (ethyl acetate extract 300mg/kg treated).

Preparation of extract dose:
2% acacia suspension was prepared by suspending 2 gram of accurately weighed acacia powder in 100 ml of 0.9% saline. 20 ml of vehicle was taken separately to which 2 gram of dried extract was added and sonicated, this produce suspension of 100 mg/ml strength. Both ethanolic extract and ethyl acetate extract suspension were prepared in such manner. 19  • The rectal temperature each animal was recorded 15 minute before and at the interval of 30 minute after treatment using telethermometer upto 4 hours. 19

Statistical analysis
All the values of Body temperature were expressed as mean±standard error of mean (S.E.M.) and analyzed by ANOVA followed by Dunnett's test. Differences between groups were considered significant at P < 0.01 and P < 0.05 levels. Lantana camara Linn leaves powder shows the presence of fibres, xylem vessel, covering trichomes, calcium oxalate crystal, collenchymas, and parasitic stomata shown in Figure No.

3.
The leaves of Lantana camara Linn was collected and analysed the various standardization parameters.
Physiochemical parameters of the leaves of lantana camara Linn are tabulated in Concerning on the Antipyretic activity which was carried out on albino rabbits of either sex, Typhoid vaccine induce "pyrexia" in rabbits due to presence of lipopolysaccrides in it which acts as pyrogens and elevate the body temperature which was treated by paracetamol 100mg/kg, Lantana camara ethanolic and ethyl acetate extracts 300mg/kg. LC IJBAR (2011) 02(08) www.ijbar.ssjournals.com ethanolic and ethyl acetate extract start lowering the body temperature from 1.5 th hour, while between 2 nd and 3 rd hour both the extract showed significant (P<0.01) Antipyretic activity when compared with the negative control group by one way ANOVA followed by dunnett's t-test.

Conclusion and Future Prospects
Preliminary phytochemical as well as various aspects of the leaves sample were studied and described along with physico chemical and TLC studies in authentification adultration for quality control of raw drugs. The leaves of Lantana camara Linn exhibits a set of diagonistic characters which will help to identify the drug in dried condition. It has been concluded from this study that estimation is highly essential for raw drugs or plant parts used for the preparation of compound formulation drug. The periodic esessment is essential for quality assurance and safer use of herbal drugs.
The antipyretic activity of Lantana camara could be at least in part due to COX-1, COX-2 enzyme inhibition and free radical-scavenging activities which may be attributed to the presence of flavonoids and other polyphenols in the extracts. The results of this study provided a scientific support for the use of Lantana camara in the treatment of Pyrexia.