Pharmacognosy Practical Lab Manual PDF

PRACTICAL LAB MANUAL PHARMACOGNOSY D. Pharm Ist Year Sr. No Name of Experiment 1. To study the compound microscope 2. To study the macroscopic and microscopic study of the Senna Leaf. 3. To study the macroscopic and microscopic st...

PRACTICAL LAB MANUAL PHARMACOGNOSY D. Pharm Ist Year Sr. No Name of Experiment 1. To study the compound microscope 2. To study the macroscopic and microscopic study of the Senna Leaf. 3. To study the macroscopic and microscopic study of the Datura Leaf. 4. To study the macroscopic and microscopic study of the ISAPGOL 5. To study the macroscopic and microscopic study of the CARDAMOM 6. To study the macroscopic and microscopic study of the FENNEL 7. To study the macroscopic and microscopic study of the DILL 8. To study the macroscopic and microscopic study of the CORIANDER 9. To study the macroscopic and microscopic study of the Clove 10. To study the macroscopic and microscopic study of the CINNAMOM 11. To study the macroscopic and microscopic study of the GINGER 12. To study the test of the AGAR: 13. To study the test of the acacia 14. To study the test of the TRAGACANTH 15. To study the test of the GELATIN 16. To study the test of the STARCH 17. To study the test of the HONEY 18. To study the test of the CASTOR OIL  EXPERIMENT NUMBER -1 AIM: To study the compound microscope REQUIREMENTS: Compound Microscope, Permanent slide (or any other specimen). Body tube Gourse adjustment l‹nob Nose piece Objective Fine lens adjustment screw Stage Condens Pillar er s Bas e 1.1: Compound microscope Theory: The microscope is one of the most commonly used instruments in medical, paramedical and clinical laboratories. It is used to study Cell Morphology, Histology, Histopathology and Microbiology. A microscope helps us to see microscopic objects that are too small and invisible to the naked eye. Description of Compound Microscope: The Compound microscope has the following main parts 1. The supporting system. 2. The focusing system. 3. The optical or magnifying system. 4. The illumination system including: a) Source of light b) Mirror c) Condenser 1. The support system: It is a framework to which various functional units are attached. It consists of the following: a) Base: it is a heavy metallic, ’U’ shaped or ’horseshoe’ shaped base with supports the microscope on work table and provides maximum stability. b) Pillars: There are two upright pillar that project up from the base and are attached to the ’C’ shaped handle. This allows the microscope to be tilted at a suitable angle for comfortable observation. c) Body tube: It is 16-17 cm long cylindrical tube fitted at the upper end of handle which is vertical or at an angle through which light passes via the eye piece to the observer's eye visualizing the image d) The stage: The stage is a square platform with an aperture in its centre and fitted to the limb below the objective lenses. When the slide is placed on it, converging rays of light emerging from the condenser passes through slide and then objective lens into the body tube. It can be either the fixed stage or the mechanical stage. The fixed stage has two clips that hold the slide in position. The mechanical stage has a calibrated metal frame fitted on right side of stage. It has a spring mounted clip to hold the slide and two screw heads to move the slide from side to side, forward and backward. The Vernier scale is also attached to indicate degree of movement The focusing system: The focusing system consists of coarse and fine adjustment and screw heads are used for raising and lowering body tube for proper focusing the slide. The coarse adjustment moves the focusing system up or down through a large distance via a rack. The fine adjustment works in same way which requires several rotations to move the tube through a small distance. It is employed for accurate focusing. The optical or magnifying system: It consists of body tube, eyepiece and the nosepiece. The body tube is the present between upper end of objective and eyepiece. The eyepiece fits into top of body tube. They can be 5X, 6X, 8X, 10X or 15X. Each eyepiece has two lenses; the eye lens at the top and field lens at the bottom. The field lens collects divergent rays, passes through eye lens to further magnify the image. The Nosepiece has two parts; the fixed nosepiece and the revolving nosepiece. The fixed nosepiece holds the revolving nosepiece that carries interchangeable objective lenses. The objective lenses are spring loaded objectives of different magnifying powers. Different types of the objective lenses are low power objective or 10X, high power objective or 45X, oil İmmersion objectives Of 100X and scanning objective 3X. Magnification: Magnification is the ability to make small objects seen larger, such as making a microscopic organism visible. The objective lenses magnify the images as stated below: Low power objective 10X = 10 x 10 = 100 times. High power objective 45X = 45 x 10 = 450 times. oil immersion objective (100X) = 100 x 10 = 1000 times. oil immersion (100X) objective has very small aperture and deep focusing position i.e. 1 mm from the slide. The light rays coming from the slide (denser medium) are refracted by thin layer of air (rarer medium) away from the small aperture of the objective and results in faint image. If some other medium like cedar wood oil, paraffin or glycerin having same refractive index as that of glass is added on the slide, it removes the thin layer of air and forms a continuous medium. This avoids the refraction of light rays and results in sharp image. 2. The illumination system: The microscope will function only when proper illumination or lightning is provided. The illumination system is to provide uniform, soft and bright illumination. The illumination system consists of: (a)Source of light: It may be external (natural day light, electric lamp or tube light) or internal (electric in- built light source). (b)A condenser: It is a system of lenses filled as short cylinder mounted below the stage. (r) Mirror: A double sided mirror with one flat side and other concave is located below condenser and can be rotated in all directions. It focuses light rays into a solid cone of light onto the material under study and helps in resolving image. (d) Iris diaphragm: Iris diaphragm is a thin opaque membranous structure fitted within condenser with a small lever on the side. The lever can adjust size of aperture of diaphragm and allows less or more light falling on slide, Procedure: 1. Examine the permanent slide/blood film/specimen first with naked eye. 2. Place the microscope on working table in an upright position, and raise the body tube approximately 7- 8 cm above the stage. Put the slide on the stage and using the mechanical stage, bring the specimen over the central aperture. 3. Select the low magnification objective (10X). ‘ 4. Select and adjust the mirror (plane or concave) so that the light shines on the specimen 5.Adjust the condenser well down, and partly close the diaphragm to cut down excess light. 6. Looking from the side, and using the coarse adjustment, brings the body tube down so that the low power lens is about 1 cm above the slide. Look into eyepiece and gently raise the tube till the slide tomes into focus. 7. Then choose the area of interest for viewing it under higher magnifications. 8. For focusing under high magnification, simply rotate the nosepiece so that the high magnification objective (45X) ’clicks’ into position. Raise the condenser to mid position and open the diaphragm to admit enough light. Use fine adjustment as required. 9. For focusing under oil immersion objective (1O0X}, raise the body tube 8-10 cm above De slide. Place a drop of cedar wood oil, paraffin or glycerin on &e slide. Looking from the side bring down the objective till it just enters the oil drop. Use other adjustment as required.  EXPERIMENT NUMBER -2 AIM: To study the macroscopic and microscopic study of the Senna Leaf. SYNONYMS: Senai ki patti, Sonamukhi, Tinnevelly senna. BIOLOGICAL SOURCE: It consists of dried leaflet of cassia angustifolia Vahl. Belonging to family Leguminosae and contained not less than 2.0% of glycosides calculated as Sennoside B. Macroscopy: Organoleptic CHARECTERS: Colour: Yellowish, Odour: slight, Taste: Mucilaginous, slightly bitter. EXTRA FEATURES: Surface: Isobilateral, thin, pubescent (hairy) with trichromes on both surfaces. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL REAGENTS OBSERVATIO CHARECTERISTI NO N CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle),sclerenchyma (pericycle) 2 Ruthenium red RED/PINK Mucilaginous cells of epidermis 3. Sudan red III pink Cutin/cuticle 4. Acetic acid Crystal insoluble Calcium oxalate 5. Hydrochloric acid Crystal soluble Calcium oxalate 6 Sulphuric acid (60%) Crystal soluble Calcium sulphate needles MICROSCOPIC CHARECTERISTICS OF THE POWDER DRUGS: CHEMICAL TEST: BRONTRAGER TEST FOR ANTHRAQUINONE: boil the leaves with dil.sulphuric acid. Filter and cool the filtrate. Add immiscible organic solvent layer in another test tube. add strong ammonia solution, shake slightly and keep the test tube aside , lower ammonical layer shows pink/ red colour. Chemical constituents: Mainly anthraquinone glycosides: Sennosides A, B, C, D. Uses: Irritant purgative.  EXPERIMENT NUMBER -3 AIM: To study the macroscopic and microscopic study of the Datura Leaf. Synonym: Datura herb Biological Source: Datura mainly consists of the dried leaves an dflowering tops of the Datura metel Family : Solanaceae. It contains not less than 0.2% of total alkaloids calculated as Hyoscyamine. ORGANOLEPTIC CHARECTERISTIC: Colour: Pale green, odour: disagreeable characteristic Taste: unpleasant bitter. EXTRA FEATURES: Texture: Thin and minutely hairy upper epidermis darker than lower, midrib prominent on lower surface.  STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL REAGENTS OBSERVATIO CHARECTERISTI NO N CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2. Dill Acetic acid insoluble Calcium oxalate 3. Sulphuric acid (60%) Crystal soluble Calcium sulphate needles CHEMICAL TESTS: 1. Vitali Morin reaction: The tropane alkaloid is treated with fuming nitric acid, followed by evaporation to dryness. Addition of methanolic pottassiu hydroxide solution to an acetone solution of nitrated residue, Violet colour is developed. 2. On addition of silver nitrate solution to solution of hyoscine hydrobrobide, yellowish white precipitate is formed which is insoluble on nitric acid, but soluble in dil. Ammonia solution. CHEMICAL CONSTITUENTS: Upto 0.5% of total alkaloids Scopolamine (hyoscine) is the main alkaloid. Hyoscyamine and atropine are present in minor quantities. USES: Parasympatholytic with anticholinergic and CNS depressant effect. Also used as mydriatic , antispasmodic and cerebral sedative. Hyoscine hydrobromide is used in motion sickness, gastric or duodenal ulcer.  EXPERIMENT NUMBER -4 AIM: To study the macroscopic and microscopic study of the ISAPGOL Synonym: Isapghula, Isapgol, Indian Psyllium Biological Source: Dried seeds of Planago ovate Forsk Family: Plantaginnaceae. ORGANOLEPTIC CHARECTERS: Colour: Pinkish grey to brown, odour : none, taste : Mucilaginous. EXTRA FEATURES: Testa is hard, translucent and smooth, Weight of 100 seeds = 0.15-0.19 gms, Swelling factor= 10.25-13.50 STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATI CHARECTERISTI ON CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2. Ruthenium Red Red Mucilage present in epidermis 3. Alcoholic Picric acid Yellow Aleurone grains present in the cells of endosperm and embryo 4 Sudan Red III Red Oil globules present in the cells of endosperm and embryo CHEMICAL TESTS: TEST OBSERVATION INFERENCE Seeds on slide + water Zone of mucilage is formed Mucilage present around each seed Powder+ Water Forms jelly like Mucilage present mass CHEMICAL CONSTITUENTS: Mucilage 10%, carbohydrates, fixed oil, proteins. USES: demulcent, laxative in chronic constipation. Mucilage is used as a coating materials binding agent etc.  EXPERIMENT NUMBER -5 AIM: To study the macroscopic and microscopic study of the CARDAMOM Synonym: Chotti - Ilaychi Biological Source: It consists of dried, nearly ripe fruit of Elettaria cardamomum Var. Family: Zingiberaceae. Seeds contain of less than 4% of volatile oil. ORGANOLEPTIC CHARECTERS: Colour: Pericarp,: Green to pale buff, Seeds: pale to reddish brown , odour: strongly aromatic and characteristic, taste: strongly aromatic. EXTRA FEATURES: fruits contain three chambers, each consists of two rows of seeds. Seeds are enclosed in membranous arillus. Each chamber contains six to ten seeds in two rows. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERISTI CS 1. Phloroglucinol+ Conc. HCl(1:1) RED/PINK Lignified tissues: xylem(vascular bundle)) 2. SULPHURIC ACID (60%) RED Calcium oxalate 3. Dil. Iodine solution blue Starch present in perisperm 4 Sudan Red III red Oil globule 5 Alcoholic picric acid yellow Aleurone grains CHEMICAL CONSTITUENTS: Volatile oils: Cineol, limonene, borneol, L-terpenol, starch, fixed oil, silica. USES: carminative, stimulant, aromatic, flavouring agent  EXPERIMENT NUMBER -6 AIM: To study the macroscopic and microscopic study of the FENNEL Synonym: Bari sauf Biological Source: Consists of dried ripe fruit of cultivated spices Foeniculum vulgare. It contains not less than 1.4% of volatile oil. Family: Umbelliferae. ORGANOLEPTIC CHARECTERS: Colour: Greenish or yellowish brown Odour: sweet aromatic and characteristic, Taste: Sweet, mucilaginous, agreeable, aromatic and characteristic. EXTRA FEATURES: fennel exhibits cremocarp. It consists of two equal portions called as Mericarps , connected by central stalks called as carpophores. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERISTI CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2 Sudan Red III red Oil globule in the cells of endosperm and cuticle 3 Alcoholic picric acid yellow Aleurone grains CHEMICAL CONSTITUENTS: Volatile oils: (4-6%), Anethole 50-60% of vol. oil, d- fenchone 10% of volatile oil, Fixed oil 12-18%, Proteins (14-22%) Uses: carminative, respiratory stimulant, aromatic.  EXPERIMENT NUMBER -7 AIM: To study the macroscopic and microscopic study of the DILL Synonym: Fructus anethi, European dill, Anethum Biological Source: Consists of dried ripe fruit of Anethum graveolens. It contains not less than 2.5% of volatile oil. Family: Umbelliferae. ORGANOLEPTIC CHARECTERS: Colour: chocolate brown, Odour: Aromatic, spicy and characteristic, Taste; Spicy, aromatic, and characteristic. EXTRA FEATURES: normally separated mericarps, dorsally compressed. Orthospermous fruits. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERISTI CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2 Sudan Red III red Oil globule in the cells of endosperm and cuticle 3 Alcoholic picric acid yellow Aleurone grains CHEMICAL CONSTITUENTS: volatile oils 3-4%, 50-60% carvone , 5% dihydrocarvone , D- limonene and phellandrene etc, fixed oil protein. USES: Aromatic, stimulant, Antiseptic.  EXPERIMENT NUMBER -8 AIM: To study the macroscopic and microscopic study of the CORIANDER Synonym: Dhania (hindi) Biological Source: Consists of dried ripe fruit of Coriandrum sativum. It contains not less than 0.3% of volatile oil. Famil : Umbelliferae. ORGANOLEPTIC CHARECTERS: Colour: Brownish yellow, Odour: Aromatic, taste: Spicy and characteristic. EXTRA FEATURES: Cremocarps, caelospermous fruit. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERISTI CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2 Sudan Red III red Oil globule in the cells of endosperm and cuticle 3 Alcoholic picric acid yellow Aleurone grains MICROSCOPICAL CHARECTERISTICS OF POWDERED DRUG: CHEMICAL CONSTITUENTS: Volatile oil 0.2-1%: coriandrol (D-Linalool 60-70%). Terpene 20%, Fixed oil 13-20%, Proteins 17%. USES: carminative, aromatic, stimulant, spice, flavouring agent.  EXPERIMENT NUMBER -9 AIM: To study the macroscopic and microscopic study of the Clove Synonym: Lavang (Hindi) Biological Source: It consists of dried flower buds of Eugenia caryophyllum. It contains not less than16% of Clove oil. Family: Myrtaceae. ORGANOLEPTIC CHARECTERS: Colour: Dark brown or crimson red, Odour: Aromatic, Taste; Spicy pungent followed by numbness. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERIST ICS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2 Sudan Red III red Oil globule in the cells of endosperm and cuticle 3 Strong KOH solution Needle shaped Eugenol of Volatile potassium eugenate oil crystals 4 Dil hydrochloric acid Crystals soluble Calcium Oxalate crystals 5 Sulphuric acid Soluble , needles of Calcium calcium sulphate on oxalate crystals standing CHEMICAL TEST: SL NO TESTS OBSERVATION INTERFERENCE 1 Aqueous extract+ Lead acetate White ppt Tannins solution 2 Clove oil + alcohol+ ferric chloride Blue colouration Eugenol 5% solution 3 Aqueous extract + Ferric chloride Dark colour tannins solution5% CHEMICAL CONSTITUENTS: volatile oil: Eugenol, isoeugenol, methyl and dimethyl furfural, alpha and beta caryophylline , hydrolysable tannins. USES: Carminative, aromatic, antiseptic, stimulant, flavouring agent, dental analgesic oil, in microscopic work , for isolation of eugenol.  EXPERIMENT NUMBER -10 AIM: To study the macroscopic and microscopic study of the CINNAMOM Synonym: Cinnamon bark, kalmi-dalchini Biological Source: It Consists of dried inner bark of the shoots of coppied trees of Cinnamomum zelanicum,It contains not less than 0.1% volatile oil. Family: Lauraceae. ORGANOLEPTIC CHARECTERS: Colour: outer surface dull yellowish brown, inner surface darker in colour, Odour: Fragrant, Taste: Warm, sweet. EXTRA FEATURES: Bark is free of cork, single or double quillsor or single closely packed compound quill. Fracture: splintery. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERISTI CS 1. Phloroglucinol+ Conc. HCl(1:1) RED/PINK Lignified tissues: xylem(vascular bundle)) 2 Iodine blue strach 3 Ruthanium red pink Mucilage cells 4 Dil hydrochloric acid Crystals soluble Calcium Oxalate crystals 5 Accetic acid insoluble Calcium oxalate crystals 6 1% solution of osmic acid Brown or pale Volatile oil 7 Dil. Tincture alkana Red on standing Volatile oil 30mins CHEMICAL TEST: SL NO TESTS OBSERVATION INTERFERENCE 1 Volatile oil+ 5ml of alcohol+ one Green colour Cinnamic aldehyde + drop of ferric chloride eugenol present in volatile oil 2 Chloroform extract or volatile oil Red shaped crystals Cinnamic aldehyde on slide+ drop of 10% aqeous phenyl hydrazine hydrochloride solution 3 Aq. Solution + FeCl3 solution Dark colour Tannin 4 Aq. Extract + lead accetate White ppt Tannin 5 AQ. Extract + Decolourisation Tannin potassium permanganate solution CHEMICAL CONSTITUENTS: volatile oil 0.5-1%:Cinnamic aldehyde (55-65%) , Eugenol, tannins, starch ,, calcium oxalate. USES: Carminative, aromatic, mild astringent, powerful germicide.  EXPERIMENT NUMBER -11 AIM: To study the macroscopic and microscopic study of the GINGER Synonym: Sonth, zinger, Jamica ginger Biological Source: It Consists of dried rhizome of Zingiber officinale , scrapped to remove the dark outer skin and dried in the sun. Family: Zingiberaceae. ORGANOLEPTIC CHARECTERS: Colour: externally buff colour, odour: agreeable and aromatic, Taste: agreeable, pungent, and characteristic. EXTRA FEATURE: Sympodial branching, horizontal rhizome, transversely cut surface shows well marked endodermis and stele. Fracture: short and fibrous. STRAINING/ DIAGNOSIS/MACRO-CHEMICAL TESTS: SL NO REAGENTS OBSERVATION CHARECTERISTI CS 1. Phloroglucinol+ Conc. RED/PINK Lignified tissues: HCl(1:1) xylem(vascular bundle)) 2 Iodine blue starch CHEMICAL TEST: Boil the drug with 5% potassium hydroxide or alkali – pungency of the ginger destroyed. CHEMICAL CONSTITUENTS: volatile oil, Terpenes, cineol, citral, borneol, pungent principle: gingerol, shogaol, zingerone. USES: Carminative, stimulant, flavouring agent  EXPERIMENT NUMBER -12 AIM: To study the test of the AGAR: CHEMICAL REQUIREMENTS: agar, ruthenium red, barium chloride, fehling solution, iodine solution. Test of Agar: Sl. NO Experiments Observation Inference 1 Agar is boiled with water Stiff jelly on Agar may cooling be present 2 Agar solution is treated Pink color Agar may with ruthenium red be present 3 Hot agar solution is White PPT Agar may treated with barium be present chloride reagent 4 Hot agar solution is Red PPT Agar may treated with fehling’s be present solution and heated 5 Agar solution is treated Crimson to Agar may with iodine solution brown color be present 6 Agar ash taken on a slide Spongy Agar may and add two drops HCl spicules of be present and observed under diatoms are microscopes observed  EXPERIMENT NUMBER -13 AIM: To study the test of the acacia CHEMICAL REQUIREMENTS: Acacia, Strong lead acetate, Sulphuric acid, Benzedrine, Iodine solution, HCl, Fehling solution Test for Acacia: SL NO EXPERIMENTS OBSERVATION INFERENCE 1 5ml of a 2%W/V Flocculent white Acacia may solution of acacia is PPT is produced be present treated with 1 ml of strong lead sub acetate solution 2 Dissolve 0.25gm of Unstable deep Acacia may acacia in 5 ml of water blue color is be present by shaking in cold. add noticed (due to 0.5 ml of H2SO4 and 0.5 enzyme oxidase) ml1% solution of benzidine in alcohol, shake and allowed to stand 3 10ml of 2%W/V No ppt is Acacia may solution of acacia, add produced be present 0.2 ml of a 20%w/v solution of lead acetate 4 0.1 gm of acacia powder The mixture does Acacia may add 1 ml of N/50 iodine not acquire be present solution crimson color (distinction from agar and  tragacanth) 5 To 1 ml of acacia solution Red ppt (due to Acacia may add 4 ml of water and cuprous oxide) be present dilute HCl acid and boiled for few minutes. Add Fehling’s solution and heat  EXPERIMENT NUMBER -14 AIM: To study the test of the TRAGACANTH CHEMICAL REQUIREMENTS: Tragacanth, Ferric chloride solution, Copper oxide, conc. Ammonium hydroxide, Lead accetate, Fehling solution, Caustic potash. Test for Tragacanth: Sl no Experiment Observation Inferences 1 Drug is boiled with Deep yellow ppt Tragacanth may freshly prepared 10% be present aqueous ferric chloride solution 2 Dissolve tragacanth and Stringy ppt Tragacanth may precipitated copper be present oxide in conc. ammonium hydroxide 3 Tragacanth solution is Red ppt Tragacanth may treated with Fehling’s be present solution and heated 4 Drug solution treated White ppt Tragacanth may with lead acetate be present solution 5 Tragacanth is treated Canary yellow Tragacanth may with 5%aqueous caustic color be present potash  EXPERIMENT NUMBER -15 AIM: To study the test of the GELATIN Chemical requirements:  Gelatin  Copper sulphate  Conc. Nitric acid  Mercuric nitrate  Ninhydrin solution  Nitrous acid Sl. No Test Observation Inference 1 Biuret test: To alkaline A red or violet color solution of a protein (2ml), is obtained with a dilute solution of copper peptide containing sulphate is added at least two peptide linkage 2 Xanthoproteic Give yellow fever The color reaction:Protein+ warmed becomes orange with conc. Nitric acid color when solution made alkaline 3 Millon’s reagent:(Mercuric Give white PPT PPT turn on red nitrate in Nitric acid color on heating. containing a trace of nitrous acid) 4 Aq. Solution of protein + Red to violet color alcoholic solution of Ninhydrin and heated  EXPERIMENT NUMBER -16 AIM: To study the test of the STARCH Chemical requirements:  Starch  Iodine solution Sl. No Test Observation 1 Boil 1 gm of starch with 15 ml of The transcalent viscous water and cool jelly is produced 2 Adding iodine solution on the above Jelly turns deep blue in jelly colour. The blue colour disappears on warming and reappear on colling  EXPERIMENT NUMBER -17 AIM: To study the test of the HONEY Chemical requirements:  Honey  Fehling’s solution A and B  Molisch reagent (α- Naphthol dissolved in Ethanol)  Conc. H2SO4 Sl. No Test Observation Inference 1 Honey + Fehling Red PPT Honey is present solution A and B 2 Sugar solution + Purple ring form Honey is present Molisch reagent +conc. H2SO4  EXPERIMENT NUMBER -18 AIM: To study the test of the CASTOR OIL Chemical requirements:  Petroleum Ether Sl. No Test Observation 1 Castor oil treated with the half of its Completely soluble in volume Petroleum Ether (50-60ͦ) petroleum ether 2 Oil +equal volume of alcohol and cool to Castor oil is present 0C for 3 hrs.