Continuous Hot Percolation Process Biology Essay

Continuous Hot Percolation Process Biology studyThe collected, cleaned and coarsely powde going of Clerodendrum phlomidis (Linn) was used for the pluckion purposes. 1kg of powde personnel casualty leaves was used. It was then take outed with versatile solvents from non polar to polar such as oil semblance ether, anesthetize, ethyl radical ethanoate and Methanol. The solvents used were distilled before use. The pull oution was carried out with non-homogeneous solvents by of tropical soxhlet take awayion for 72 Hrs. after(prenominal)ward each solvent extraction, the extracts were perk uped through whattmann slobber paper to remove any impurities is present.PREPARATION OF EXTRACTSa) Petroleum ether extract of leaves of Clerodendrum phlomidis (Linn).About 1kg of dry coarse powder was extracted first 5 liters of Petroleum ether (60-80C) in a spicy soxhlet extraction using declamatory bottomed flask for 72 Hrs. After completion of extraction the crude ether was filte go ing and concentrated to dry mass by vaccum distillation. A colorful green colour residue (1.48 % w/w) was obtained. The extract was then stored in a desicator (Practical pharmacognosy. 1994).b) Chloroform extract of leaves of Clerodendrum phlomidis (Linn).The marc left after Petroleum ether extract, was dried and subsequently extracted with 4 liters of Chloroform (40-60C) in a hot soxhlet extraction using round bottomed flask for 72 Hrs. After completion of extraction, it was filtered and the solvent was removed by distillation infra cut back pressure. A yellowish green colour residue (0.86 %w/w) was obtained .The extract was then stored in a desicator.c) ethyl radical acetate extracts of leaves of Clerodendrum phlomidis (Linn).The marc left after Chloroform extract, was dried subsequently extracted with 3 liters of methanol (40-60C) in a hot soxhlet extraction using round bottomed flask for 72 Hrs. After completion of extraction, it was filtered and the solvent was removed by d istillation beneath reduced pressure. A dark-brownness colour residue (0.63 %w/w) was obtained. The extract was then stored in a desicator.d) Methanol extracts of leaves of Clerodendrum phlomidis (Linn).The marc left after Ethyl acetate extract, was dried subsequently extracted with 2 liters of methanol (40-60C) in a hot soxhlet extraction using round bottomed flask for 72 Hrs. After completion of extraction, it was filtered and the solvent was removed by distillation under reduced pressure. A dark brown colour residue (8.24 %w/w) was obtained. The extract was then stored in a desicator.From the weight of the each extractive residue, the extractive set were calculated in percentage. All the above extracts were used for identification of constituents by phytochemical outpourings and for the pharmacological studies. The yields of various extract were shown in the flurry No 1.Table No.1EXTRACTIVE VALUES OF THE LEAVES OFCLERODENDRUM PHLOMIDIS (LINN)S.NoEXTRACTSYEILD(gms)% YIELD(w/ w)1Petroleum ether14.81.482Chloroform8.60.863Ethyl acetate6.30.634Methanol82.48.24QUALITATIVE PHYTOCHEMICAL compendQualitative chemical essays were carried out for all the extracts of leaves of Clerodendrum phlomidis (Linn) to identify the various phytoconstituents. The various adjudicates and reagents used argon given below and observations are recorded. (Table No.2) examinations for carbohydratesMolishs judgeTo 2-3 ml of extract, added few drops of -naphthol result in alcohol, shaken and added concentrated H2SO4 form sides of the screen tube was observed for violet ring at the junction of cardinal liquids (Indian Pharmacopeia, vol II. 199).Fehlings rill1 ml Fehlings A and Fehlings B upshots was mixed and turn for one minute. Added equal volume of test answer. Heated in simmering water lav for 5-10 min was observed for a yellow, then brick red overhasty.Benedicts testEqual volume of Benedicts reagent and test stem in test tube were mixed. Heated in boiling water bath for 5 min. Solution may appear green, yellow or red depending on amount of reducing sugar present in test beginningTests for AlkaloidsMayers testTo the 1 ml of extract, add 1 ml of Mayers reagent (Potassium mercurous iodide firmness of purpose). Whitish yellow or cream colored precipitate indicates the posture of alkaloids.Dragendroffs testTo 1 ml of the extract, add 1 ml of Dragendroffs reagent (Potassium bismuth iodide radical). An orange-red precipitate indicates the nominal head of alkaloids.Hagers testTo 1 ml of the extract, add 1 ml of Hagers reagent (saturated aqueous dissolvent of picric acid). A yellow colored precipitate indicates the presence of alkaloids.Wagners testTo 1 ml of the extract, add 1 ml of Wagners reagent (Iodine in potassium iodide etymon). Formation of reddish brown precipitate indicates the presence of alkaloids. (Kokate C.K et.al, 2007).Tests for GlycosidesHydrolysis of extractA minimum quantity of the extracts is hydrolyzed with hydrochloric ac id for few legal proceeding on water bath and the hydrolysate is subjected to the following tests.). Legals testTo the hydrolysate 1 ml pyridine and few drops of sodium nitropruside solution added, then it is made alkaline with sodium hydroxide solution. Color change shows the presence of glycosides.). Borntragers testHydrolysate is treated with anaesthetize and the anaesthetise layer is separated. To this, equal quantity of dilute ammonia solution is added. Color changes in the ammonical layer shows the presence of glycosides.Bal jets testA test solution observed for yellow to orange color with sodium picrate.Keller Killiani test melt down the extract in acetic acid containing traces of ferric chloride and transplant to a test tube containing sulphuric acid. At the junction, organization of a reddish brown color, which gradually becomes low-spirited, confirms the presence of glycoside.Tests for Phyto SteroidsSmall quantity of extract is dissolved in 5 ml of anesthetize separa tely. The above obtained chloroform solutions are subjected to Salkowski and Liebermann Burchard tests (Harbone. JB. 1973).Salkowski testTo the 1 ml of above prepared chloroform solution few drops of concentrated sulphuric acid is added. Formation of brown ring indicates the presence of phytosterols.Liebermann Burchard testThe above prepared chloroform solutions are treated with few drops of concentrated sulphuric acid followed by 1 ml of acetic anhydride solution. A bluish green color solution shows the presence of phytosterols.Tests for FlavanoidsShinoda testTo dried powder or extract added 5 ml 95% ethanol, few drops concentrated HCl and 0.5 g magnesium turnings. knap color was observed (Quality Control of Herbal Drugs. 2002).Ferric Chloride testTest solution with few drops of ferric chloride solution shows intense green color. alkalic reagent testTest solution when treated with sodium hydroxide solution shows increase in the intensity of yellow color, which becomes colourless on admittance of drops of dilute acid.Lead Acetate solution testTest solution with few drops of lead acetate solution (10%) gives yellow precipitates.Test for terpenoidsDissolve 2 to 3 granules of tin metal in 2 ml of thionyl chloride solution. Then add 1 ml of the extract into the test tube. The formation of a pink color indicates the presence of terpenoids.5 ml of aqueous extract of each plant sample is mixed with 2 ml of CHCl3 in a test tube. 3 ml of concentrated H2SO4 is conservatively added to the mixture to form a layer. An interface with a reddish brown coloration is formed if terpenoids constituent is present. (Journal of Medicinal Plants Research Vol. 3(2), pp.068).Tests for SaponinsFoam testThe extracts are diluted with 20ml of distilled water and then agitated in a graduated cylinder for 15minutes. Formation of foam layer indicates the presence of saponins. (Khandelwal K.R, 2007). haemolytic testAdded test solution to one drop of blood rigid on glass slide. Haemolytic zone whether appeared was observed.Tests for Proteins and Amino acidsBiuret testTo 3 ml test solution added 4% NaOH and few drops of 1% CuSO4 solution observed for violet or pink color (Practical Pharmacognosy. 1996).Millions test complicated 3 ml test solution with 5 ml Millions reagent, white-hot precipitate. Precipitate warmed turns brick red or precipitate dissolves giving red color was observed.Xanthoprotein testMixed 3 ml test solution with 1 ml concentrated H2SO4 observed for white precipitate.Ninhydrin test3 ml test solution and 3 drops 5% Ninhydrin solution were het up in boiling water bath for 10 min. observed for imperial or bluish colorTests for Tannins and Phenolic compoundsTo 2 3 ml of extract, add few drops of following reagents5% FeCl3 solution deep blue black color.Lead acetate solution white precipitate.Gelatin solution white precipitate.Bromine water decoloration of bromine water.Acetic acid solution red color solution.Dilute iodine solution transient red co lor.Dilute HNO3 reddish to yellow color.Test for Fixed Oils and FatsSpot testSmall quantity of the extract is placed between two filter papers. Oil stain produced with any extract shows the presence of fixed oils and fats in the extracts.Saponification testFew drops of 0.5N alcoholic potassium hydroxide are added to the extract with few drops of phenolphthalein solution. Later the mixture is heated on water bath for 1 2 hours soap formation indicates the presence of fixed oils and fats in the extracts.Test for Gums and MucilagesRuthenium red testSmall quantities of extract are diluted with water and added with ruthenium red solution. A pink color production shows the presence of gums and mucilages.TABLE NO 2QUALITATIVE PHYTOCHEMICAL psychoanalysis OF EXTRACTS OF LEAVES OF CLERODENDRUM PHLOMIDIS (LINN)TEST OF EXTRACTSPETROLEUMETHEREXTRACTSCHLOROFORMEXTRACTSETHYL ACETATEEXTRACTS wood spiritEXTRACTSCARBOHYDRATES__++ALKALOIDS__++GLYCOSIDES++++PHYTO STEROIDS++++FLAVONOIDS_+++TERPENOIDS ++++SAPONIN_+++TANNINS PHENOLIC COMPOUNDS++++FIXED OILS FATS+_++GUMS MUCILAGES_+++PROTEINS AMINO ACIDS_+++(+ ) = indicates presence, (-) = indicates absenceBased on soft analysis we have selected Ethyl acetate extract of clerodendrum phlomidis (Linn) leaves for further studies because Ethyl acetate extract is having more phytoconstituents when compared to all other extracts.