The collected, cleaned and coarsely powdered of Clerodendrum phlomidis ( Linn ) was used for the extraction intents. 1kg of powdery foliages was used. It was so extracted with assorted dissolvers from non polar to polar such as Petroleum quintessence, Chloroform, Ethyl ethanoate and Methanol. The dissolvers used were distilled earlier usage. The extraction was carried out with assorted dissolvers by of hot soxhlet extraction for 72 Hrs. After each dissolver extraction, the infusions were filtered through whattmann filter paper to take any drosss is present.
Preparation OF EXTRACTS
Petroleum ether infusion of foliages of Clerodendrum phlomidis ( Linn )
About 1kg of dry coarse pulverization was extracted foremost 5 litres of Petroleum quintessence ( 60-80A°C ) in a hot soxhlet extraction utilizing unit of ammunition bottomed flask for 72 Hrs. After completion of extraction the crude oil quintessence was filtered and concentrated to dry mass by vaccum distillment. A dark green coloring material residue ( 1.48 % w/w ) was obtained. The infusion was so stored in a desicator ( Practical pharmacognosy. 1994 ) .
Chloroform infusion of foliages of Clerodendrum phlomidis ( Linn )
The marc left after Petroleum ether infusion, was dried and later extracted with 4 litres of Chloroform ( 40-60A°C ) in a hot soxhlet extraction utilizing unit of ammunition bottomed flask for 72 Hrs. After completion of extraction, it was filtered and the dissolver was removed by distillment under decreased force per unit area. A xanthous green coloring material residue ( 0.86 % w/w ) was obtained.The infusion was so stored in a desicator.
Ethyl acetate infusions of foliages of Clerodendrum phlomidis ( Linn )
The marc left after Chloroform infusion, was dried later extracted with 3 litres of methyl alcohol ( 40-60A°C ) in a hot soxhlet extraction utilizing unit of ammunition bottomed flask for 72 Hrs. After completion of extraction, it was filtered and the dissolver was removed by distillment under decreased force per unit area. A brown coloring material residue ( 0.63 % w/w ) was obtained. The infusion was so stored in a desicator.
Methanol infusions of foliages of Clerodendrum phlomidis ( Linn )
The marc left after Ethyl ethanoate infusion, was dried later extracted with 2 litres of methyl alcohol ( 40-60A°C ) in a hot soxhlet extraction utilizing unit of ammunition bottomed flask for 72 Hrs. After completion of extraction, it was filtered and the dissolver was removed by distillment under decreased force per unit area. A dark brown coloring material residue ( 8.24 % w/w ) was obtained. The infusion was so stored in a desicator.
From the weight of the each extractive residue, the extractive values were calculated in per centum. All the above infusions were used for designation of components by phytochemical trials and for the pharmacological surveies. The outputs of assorted infusion were shown in the
QUALITATIVE PHYTOCHEMICAL ANALYSIS
Qualitative chemical trials were carried out for all the infusions of foliages of Clerodendrum phlomidis ( Linn ) to place the assorted phytoconstituents. The assorted trials and reagents used are given below and observations are recorded.
Trials for saccharides:
- Molish ‘s trial: To 2-3 milliliter of infusion, added few beads of I±-naphthol solution in intoxicant, shaken and added concentrated H2SO4 signifier sides of the trial tubing was observed for violet ring at the junction of two liquids ( Indian Pharmacopeia, vol II. 199 ) .
- Fehling ‘s trial: 1 milliliter Fehling ‘s A and Fehling ‘s B solutions was assorted and boiled for one minute. Added equal volume of trial solution. Heated in boiling H2O bath for 5-10 min was observed for a yellow, so brick ruddy precipitate.
- Benedict ‘s trial: Equal volume of Benedict ‘s reagent and trial solution in trial tubing were assorted. Heated in boiling H2O bath for 5 min. Solution may look green, xanthous or ruddy depending on sum of cut downing sugar nowadays in trial solution
Trials for Alkaloids
- Mayer ‘s trial: To the 1 milliliter of infusion, add 1 milliliter of Mayer ‘s reagent ( Potassium mercuric iodide solution ) . Milky yellow or pick colored precipitate indicates the presence of alkaloids.
- Dragendroff ‘s trial: To 1 milliliter of the infusion, add 1 milliliter of Dragendroff ‘s reagent ( Potassium bismuth iodide solution ) . An orangish-red precipitate indicates the presence of alkaloids.
- Hager ‘s trial: To 1 milliliter of the infusion, add 1 milliliter of Hager ‘s reagent ( saturated aqueous solution of picric acid ) . A xanthous coloured precipitate indicates the presence of alkaloids.
- Wagner ‘s trial: To 1 milliliter of the infusion, add 1 milliliter of Wagner ‘s reagent ( Iodine in K iodide solution ) . Formation of ruddy brown precipitate indicates the presence of alkaloids. ( Kokate C.K et.al, 2007 ) .
Trials for Glycosides
Hydrolysis of infusion:
A minimal measure of the infusions is hydrolyzed with hydrochloric acid for few proceedingss on H2O bath and the hydrolysate is subjected to the undermentioned trials.
- Legal ‘s trial: To the hydrolysate 1 milliliter pyridine and few beads of Na nitropruside solution added, so it is made alkaline with Na hydrated oxide solution. Color alteration shows the presence of glycosides.
- Borntrager ‘s trial: Hydrolysate is treated with trichloromethane and the trichloromethane bed is separated. To this, equal measure of dilute ammonium hydroxide solution is added. Color alterations in the ammonical bed shows the presence of glycosides.
- Bal jet ‘s trial: A trial solution observed for yellow to orange colour with Na picrate.
- Keller Killiani trial: Dissolve the infusion in acetic acid incorporating hints of ferrous chloride and transportation to a trial tubing incorporating sulfuric acid. At the junction, formation of a ruddy brown colour, which bit by bit becomes bluish, confirms the presence of glycoside.
Trials for Phyto Steroids
Small measure of infusion is dissolved in 5 milliliter of trichloromethane individually. The above obtained chloroform solutions are subjected to Salkowski and Liebermann – Burchard trials ( Harbone. JB. 1973 ) .
- Salkowski trial: To the 1 milliliter of above prepared trichloromethane solution few beads of concentrated sulfuric acid is added. Formation of brown ring indicates the presence of phytosterols.
- Liebermann – Burchard trial: The above prepared chloroform solutions are treated with few beads of concentrated sulfuric acid followed by 1 milliliters of acetic anhydride solution. A blue green colour solution shows the presence of phytosterols.
Trials for Flavanoids
- Shinoda trial: To dried pulverization or infusion added 5 ml 95 % ethyl alcohol, few beads concentrated HCl and 0.5 g Mg turnings. Pink colour was observed ( Quality Control of Herbal Drugs. 2002 ) .
- Ferric Chloride trial: Test solution with few beads of ferrous chloride solution shows intense green colour.
- Alkaline reagent trial: Test solution when treated with Na hydrated oxide solution shows addition in the strength of xanthous colour, which becomes colourless on add-on of beads of dilute acid.
- Lead Acetate solution trial: Test solution with few beads of lead acetate solution ( 10 % ) gives xanthous precipitates.
Trial for terpenoids
Dissolve 2 to 3 granules of Sn metal in 2 milliliter of thionyl chloride solution. Then add 1 milliliter of the infusion into the trial tubing. The formation of a pink colour indicates the presence of terpenoids.
5 milliliter of aqueous infusion of each works sample is assorted with 2 milliliters of CHCl3 in a trial tubing. 3 milliliter of concentrated H2SO4 is carefully added to the mixture to organize a bed. An interface with a ruddy brown colour is formed if terpenoids component is present. ( Journal of Medicinal Plants Research Vol. 3 ( 2 ) , pp.068 ) .
Trials for Saponins
- Foam trial: The infusions are diluted with 20ml of distilled H2O and so agitated in a calibrated cylinder for 15minutes. Formation of froth bed indicates the presence of saponins. ( Khandelwal K.R, 2007 ) .
- Hemolytic trial: Added trial solution to one bead of blood placed on glass slide. Haemolytic zone whether appeared was observed.
Trials for Proteins and Amino acids
To 3 milliliters test solution added 4 % NaOH and few beads of 1 % CuSO4 solution observed for violet or tap colour ( Practical Pharmacognosy. 1996 ) .
Million ‘s trial:
Assorted 3 ml trial solution with 5 milliliters Million ‘s reagent, white precipitate. Precipitate warmed bends brick ruddy or hasty dissolves giving ruddy colour was observed.
Assorted 3 ml trial solution with 1 milliliters concentrated H2SO4 observed for white precipitate.
3 ml trial solution and 3 beads 5 % Ninhydrin solution were heated in boiling H2O bath for 10 min. observed for purple or blue colour
Trials for Tannins and Phenolic compounds
To 2 – 3 milliliter of infusion, add few beads of following reagents:
5 % FeCl3 solution: deep blue – black colour.
Lead acetate solution: white precipitate.
Gelatin solution: white precipitate.
Bromine H2O: decoloration of bromine H2O.
Acetic acerb solution: ruddy colour solution.
Dilute iodine solution: transient ruddy colour.
Dilute HNO3: reddish to yellow colour.
Trial for Fixed Oils and Fats
- Topographic point trial: Small measure of the infusion is placed between two filter documents. Oil discoloration produced with any infusion shows the presence of fixed oils and fats in the infusions.
- Saponification trial: Few beads of 0.5N alcoholic K hydrated oxide are added to the infusion with few beads of phenolphthalein solution. Subsequently the mixture is heated on H2O bath for 1 – 2 hours soap formation indicates the presence of fixed oils and fats in the infusions.