OF COMMERCIAL CRUDE 3 DRUGS INTENDED FOR USE * Definition of Crude drug Crude drugs are plant, animal or their parts which after collection are subjected to only drying or making them in to transverse/ longitudinal slices piece or peeling them in some cases. © Crude drug occurrence Crude drug are generally obtained by plant, animal and mineral origin. 1. Plant origin: whole plant or part of plant like leaves flowers, seeds and barks. Or vegetable saps, extracts and secretions. 2, Animal origin: whole animals, glands or organs, extracts and secretions. 3, Mineral origin: ferrous sulfate, magnesium, zinc, gold etc. Definition of drug evaluation Drug evaluation may be defined as the determination of identity, purity and quality of a drug. > Identity: identification of biological source of the drug: ry: the quantity of the active constituents present. « the extent of foreign organic material present in a crude drug. + Importance of evaluation of crude drugs > Determination of Biochemical variation in the drugs. > Identification of deterioration due treatment and storage. > Reporting Substitution and adulteration, as result of carelessness, ignorance and fraud. The evaluation of a drug is done by following methods. Organoleptic evaluation Morphological evaluation Microscopic evaluation Physical evaluation Chemical evaluation Analytical evaluation Biological evaluation Neageene3.2.4. ORGANOLEPTIC EVALUATION + This refers to drug evaluation by means of organs of sense and includes other sensory organs like color, odour, taste, size, shape and texture. © Itincludes the study of morphology and other sensory characters, A) Odour i. Distinet ii, Indistinet iii, aromatic B) Taste i, Acidic (sour) ji, Saccharine (sweet): indicates sugar or sugar like substances e.g, liquorice. iii, Saline (salty) iv. Alkaline v. Bitter: indicates presence of substances such as bitter principle eg., glycoside, alkaloids. vi. Tasteless vii, Distinctive sensations to the tongue: > Mucilaginous and oily (soft feeling) e.g., linseed. > Astringent indicates presence of tannin. > Pungent (warm biting sensation) e.g., ginger. > Acrid (irritant sensation) e.g., Aconite, coca. > Nauseous (those tending to excite vomiting) e.g. Ipecac. ©) Colour i. White: eg,, starch. ii, Pale yellow: e.g., ginger, squill, white pepper. iii, Deep yellow: eg,, peeled liquorice iv. Light pale brown: e.g., nux-vomica, fennel. y, Dark brown: e.g,, cloves bud. vi. Dark reddish brown: cinchona. vii. Red: (brick red): e.g., cinnamon bark inner portion, viii, Pale green: e.g,, lobelia. ix. Greenish brown: most of the leaf herbs. 3.2.2. MORPHOLOGICAL EVALUATION * Study of morphology includes visual examination of drug like study of shape and size of various parts of crude drugs. A) Flowers Floral parts: stigmas, corollas, anther, ovary, and receptacle.I FU A Tex! Book of Quality Control and Standardization of Herbals B) Leaves and leaflets Length, width, apex, margin, base, venation, the texture of the leaf and the hairs in upper and lower surface. The feel of the surface described as soft, hairy smooth. C) Bark The barks occur in three shapes: i. Flat or curved pieces. ii. Single quill. iii, Double quills. Barks have two surfaces, an outer and inner. The inner surface is usually lighter in color than the outer surface. 3.2.3, MICROSCOPIC EVALUATION * This method allows a more detailed examination of a crude drug and it can be used to identify organized drugs by their known histological characters. * Before examination through a microscope the material must be suitably prepared and can be done by powdering, cutting thin sections of the drug or preparing a macerate. i. Palisade Ratio Stomatal Number Stomatal Index Stomata v. Vein-islet Number vi. Vein-termination Number vii, Trichomes or plant hairs viii, Calcium oxalate crystals ix. Quantitative Microscopy: Lycopodium spore method L. Palisade ratio © It represents the average number of palisade cells beneath one epidermal cell, using four continuous epidermal cells for the count. * _ Itis determined from powdered drugs with the help of camera-Lucida. Examples: >» Adhatodavasica: 55-65 > Cassia angustifolia: 5.5-10.0 >» Digitalis lanata: 25-65 Il. Stomatal Number © The average number of stomata present per square millimeter of the epidermis is known as stomatal number.—— i 23 Examples: > Atropa belladonna: Upper epidermis: 07-10 Lower epidermis: 77-115 > Datura metel: Upper epidermis:147-160 Lower epidermis: 200-209 > Ocimum sanctum: Upper epidermis: 64-72 Lower epidermis: 175-250 IIL. Stomatal Index It is the percentage proportion of the number of stomata to the total number of epidermal cells. Stomatal number varies considerably with the age of the leaf but stomatal index is relatively constant for a given species. Stomatal index calculated by: S.1 = S/E+S Where, $.1 = Stomatal index Number of stomata per unit area E=Number of epidermal cells in the same unit area Examples: > Atropa belladonna: Upper epidermis: Nil Lower epidermis: 20.2-23.0 IV. Stomata: (primary and important function is gaseous exchange) A minute epidermal opening present on Arial parts of plants, Stomata congjsts of central pore, two kidney shaped similar cells (guard cells) & varying number of subsidiary cells. Epidermis of leaf shows different characteristics e.g. cuticle, stomata, trichomes. ‘Types of stomata i. Moss type Gymnospermous type i. Gramineous type iv. Dicotyledonous—it is having diagnostic significance and classified based on form of arrangement of subsidiary cells. Paracytic or rubiaceous or parallel-cell stomata: in this stomata two guard cells covered by two subsidiary cells, e.g. Senna . Diacytic or caryophyllaceous or cross-celled stomata: In these stomata the guard cells are covered by two subsidiary cells on right angle to that of stomata. e.g. peppermint Anisocytic or cruciferous or unequal-celled stomata: in this stomata number of guard cells is two but covered by three subsidiary cells and in that one is small in size with other two e.g. DaturaA Text Book of Quallty Control and Standardization of Herbals 4. Anomocytic or ranunculaceous or irregular- celled: In this type stoma is surrounded by varying number of subsidiary cells. e.g. digitalis e. Actinocytic or radiate-celled stomata: two guard cells are surrounded by radiating subsidiary cells. V. Vein-islet Namber Vein-islet number is defined as the number of vein-islets per sq.mm. of leaf surface. ‘Table 3.1: Examples of crude drug for vein-islet number VL. Vein-termination Number * It is defined as the number of veinlet terminations per. sq. mm of the leaf surface between midrib and margin. VII. Trichomes or plant hairs © Trichomes are the tubular elongated or glandular outgrowth of the epidermal cells Trichomes are also called as plant hairs. Trichomes consist of two parts root and body. Trichomes present in most of plant parts and are function less but sometimes perform secretory function. Depending up on the structure and the number of cells present in trichomes, they are classified in to following. i, Covering Trichomes ; ii, Glandular Trichomes . % iii, Hydathode or special Trichomes VIII. Calcium oxalate crystals © Several cell contents present in vegetable drugs. The inorganic crystalline compounds by virtue of their specific shapes can be utilized for the identification of herbal drugs. Due to this reason they are called as diagnostic characters of the plant. .. Cubical (cube shape) e.g., Senna, Glycyrrhiza. Rhombic (diamond shape) - . Tetragonal e.g,, Onion. iv. Mono clinic (all three axes are un-equal) e.g,, Gall. v. Acicular (long slender, pointed, bundles) e.g., Squill, Cinnamon. vi. Rosettes clusters (aggregation of crystals) e.g, Clove, Arjuna.25 vii, Microsphenoidal (minute in structures) e.g., Henbane. IX. Quantitative microscopy * Lycopodium spore method: it is used when especially chemical and other methods of evaluation of drugs fail to determine quality. Lycopodium spores are much characterized in shape and appearance and uniform in size (25 pm) on average, 94000 Spores present/mg of Lycopodium powder. © It consists of: i. Well defined particles which may be counted. i, Single layered cells or tissues the area of which may be traced under suitable magnification and actual area calculated iii, The objects of uniform thickness, the length of which can be measured, and actual area calculated. 3.2.4, PHYSICAL EVALUATION © Physical contents such as elasticity in fibers, viscosity of drugs containing gums, swelling factor for mucilage containing materials, froth number of saponin drugs, congealing point of volatile and fixed oils, melting and boiling points and water contents are some important parameters used in the evaluation of drugs. © Ultraviolet light is also used for determining the fluorescence of extracts of some drugs. +. Physical constants are extensively applied to the active principles of drugs, such as alkaloids, volatile oils, fixed oils etc. A few of them are: i. Moisture Content ii, Viscosity ili, Melting point iv. Solubility y. Optical Rotation vi. Refractive Index vii, Ash values viii, Extractive values ix. Volatile oil Content x. Foreign organic matter xi, swelling factor I. Moisture Content © Presence of moisture in a crude drug can lead to its deterioration due to either activation of certain enzymes or growth of microbes. © Moisture content can be determined by heating the drug at 105°C in an oven to a constant weight and calculating the loss of weight.Table 3.2: Examples of crude drug for Moisture content lo. | Drugs | Moisture Content (%w/w) 1, Aloes: Not more than 10 2. | Digitalis | Not more than 5 a Starch Not more than 15 IL. Viscosity * Viscosity of a liquid is constant at a given temperature and is an index of its composition. * Hence, it is used as a means of standardizing liquid drugs. 4 * Liquid paraffin-kinematic viscosity not less than 64-centistokes at 37.8°. * Pyroxylin-kinematic viscosity, 1100-2450 centistokes. IIL. Melting Point * It is one of the parameters to judge the purity of crude drugs containing lipids as constituents. They may of animal or plant origin and contain fixed oils, fats and waxes. The purity of the following crude drugs can be ascertained by determining their melting, points in the range shown against each of them. Table 3.3: Examples of crude drug for Melting point Sr.No.| Drugs Melting point (°C) 1. | COLOPHONY 75-85 i BEES WAX 62-65 WOOL FAT aad IV. Solubility * The presence of adulterant in a drug could be indicated by solubility studies. Table 3.4: Crude drugs and their solubility St. No. Drugs Solubility Castor oil Soluble in 3 volumes of alcohol Balsam of Peru | Soluble in chloral hydrate solution Asafoetida | Soluble in carbon disulphide Alkaloid bases | Soluble in chloroform a Colophony | Solubie in light petroleum el} e}ele}nES 27 pe v. Optical Rotation Many substances of biological origin, having a chiral centre, can rotate the plane of polarized light either to right (dextro rotatory) or to the left (laevo) The extent of rotation is expressed in degrees, plus (+) indicating rotation to the right and minus (-) indication rotation in the left. Such compounds are optically active and hence called optical rotation. ‘Table 3.5: Examples of crude drug for Optical rotation VI. Refractive Index When a ray of light passes from one medium to another medium of different density, it is bent from its original path. Thus, the ration of velocity of light in vacuum to its velocity in the substance is said to the Refractive index of the second medium. It is measured by means of refractometer. RI of a compound varies with the wavelength of the incident light, temperature and pressure. Table 3.6: Examples of crude drug for Refractive index ‘VIL. Ash values The residue remaining after incineration is the ash content of the drug. (Inorganic salts of carbonates, phosphates, silicates of sodium, potassium, calcium and magnesium) is known as ash content. Ash value is a criterion to judge the identity OR purity of the crude drug.FY A Toxt Book of Qualty Control and Standardization of Herbals] Table 3.7; Crude drugs along their Ash and acid insoluble ash value ‘Types of ash values value: Useful for detecting low grade products, exhausted products, excess of sandy and earthy matter with drug. ii Acid insoluble ash value: Used for the determination of earthy matter present on roots, rhizomes, and also on the leaves, Crude drugs contain calcium oxalate crystals the amount may vary depending on the environmental conditions. iii, Sulphated ash value: Used for the detection of low grade products. iv. Water soluble ash value: Water soluble ash value Used to detect either material exhausted by water or not (Tea leaves, Ginger rhizomes). Determination Total ash value |. Weigh accurately about 3gms of the powdered drug in a tared silica crucible. ‘i, Incinerate the powdered drug by gradually increasing the heat until free from carbon and cool. Keep it in desiccators. iii, Weigh the ash and calculate the % of the total ash with reference to the air dried sample. Determination Acid insoluble ash value i. Boil the total ash obtained as above for 5 minutes with 25ml of dilute HCL. 4 Filter and collect the insoluble matter on the ash less filter paper, wash the filter paper with hot water, ignite in tared crucible, cool and Kept in desiccators. iii Weigh the residue and calculate the acid insoluble ash of the drug. VIII. Extractive values «= In crude drugs, sometimes the active chemical constitutes cannot be determined by normal procedures. «In such cases, water, alcohol or ether soluble extractive values are determined for evaluation of such drugs. © Significances ) Useful for the evaluation especially when the constituents of the drugs cannot be readily estimated by any other means. )It also helps to indicate the nature of chemical constituents present in the drug. | ' | |©) Also helps in the identification of adulterants. «Types of extractive values ‘A. Water soluble extractive values B. Alcohol soluble extractive values C. Ether soluble extractive values 1, Water soluble extractive value © Water soluble extractive value is applied for the drugs which contain water soluble constituents such as tannins, sugars, plant acids and mucilage. Procedure i. Macerate about 5gm of the accurately weighed coarse powder with 100m of chloroform water in a 100ml volumetric flask for 24 hours. . Shake frequently for first 6 hours. . Filter rapidly through filter paper and evaporate 25ml of water. iv. Extract to dryness in a tared flat-bottomed shallow dish. v. Dry the residue at 105°C and weigh. Keep it in desiccators vi. Dry the extract to constant weight, finally, calculate the % W/W of Water soluble extractive value with reference to the air dried drug. rn Table 3.8: Crude drug and their water soluble extractive value 2. Alcohol soluble extractive values * Alcohol soluble extractive value is applied for the drugs which contain alcohol soluble constituents such as tannins, resins and alkaloids .Official method for the assay of myrth & asafoetida. © Generally, 95% ethyl alcohol is used for determination of Alcohol soluble extractive. Procedure i. Macerate about 5gm of the accurately weighed coarse powder with 100ml of 90% alcohol ina 100mI stoppered flask for 24 hours. fi. Shake frequently for first 6 hoursiii, Filter rapidly through filter paper and collect the filtrate evaporate 25ml of alcohol extract to dryness in a tared flat-bottomed shallow dish. iv. Dry the residue at 105°C and weigh. Keep it ina desiccators v. Dry the extract to constant weight; finally, calculate the % w/w of alcohol soluble extractive value with reference to the air dried drug. Table 3.9: Crude drug and their alcohol soluble extractive value 3. Ether soluble extractive value: Ether soluble extractive value is applied for the extraction of volatile oils, fixed oils and resins. * Volatile ether soluble extractive value (volatile oil). * Non volatile ether soluble extractive value (resin, fixed oils, coloring matter). Table 3.9: Crude drug and their ether soluble extractive value IX. Volatile oil content * Efficiency of several drugs is due to their odorous principle (volatile oils).Such crude drugs are standardized on the basis of their volatile oil contents. Weighed quantity of the drug is boiled with water in a round bottomed flask fitted with Clevenger apparatus. The distillate collected is graduated into volatile oil. The amount thus obtained is recorded from the tube.Es 3 Table 3.10: Volatile oil content of some crude drugs X. Foreign organic matter «The parts of the organ or organs other than those named in the definition and description of the drug are defined as foreign organic matter. «The maximum limit for the foreign organic matter is defined in the monograph of crude drug. If it exceeds the limits, deterioration in quality of the drug takes place. © The physical or Physico-chemical parameters are useful in quality profile of a crude drug evaluation. XI. Swelling Factor « — Significances i. Useful in the evaluation of crude drugs containing mucilage ii, Useful for the detection of purity of the crude drug, * Determination i, Transfer 1 gm of the seeds to a 25ml stoppered cylinder. Fill up to the 20ml mark on the cylinder with water. iii, Agitate gently and occasionally during 24 hours and allowed to stand. iv, Measure the volume occupied by the swollen seeds. 3.2.5. CHEMICAL EVALUATION Determination of the active constituent in a drug by chemical tests is referred to as chemical evaluation. «The following are various methods of chemical evaluation. > Instrumental methods > Chemical constant tests > Individual constituent chemical tests > > Micro chemical tests Preliminary chemical test © Instrumental methods: They make use of various instruments for evaluation like colorimetry, flourimetry, spectrophotometry etc. © Chemical constants tests: These are like acid value; iodine value and ester value etc are used for the identification of fixed oils and fats.22 ES * Individual chemical tests: These are the tests which are used for identifying particular drugs. * Microchemical tests: These are the tests which are carried on slides. Example: Euginol in clove oil is precipitated as potassium euginate crystals. © Preliminary chemical test: Extract obtained ising petroleum ether, chloroform, ethanol and water was prepared using the respective solvent. These extracts along with positive and negative controls were tested for the presence of active phytochemicals viz: tannins, alkaloids, phytosterols, triterpenoids, falvonoids, cardiac glycosides, anthroquinone glycosides, saponins, carbohydrates, proteins, amino acids and fixed oils & fats following standard methods. Tannins 1. Ferric chloride Test: Added a few drops of 5% ferric chloride solution to 2 ml of the test solution. Formation of blue color indicated the presence of hydrolysable tannins. 2. Gelatin Test: Added five drops of 1% gelatin containing 10% sodium chloride to 1 ml of the test solution. Formation of white precipitates confirmed the test. Alkaloids Approximately 50 mg-of extract was dissolved in 5 ml of distilled water. Further 2M hydrochloric acid was added until an acid reaction occurred and filtered. The filtrate was tested for the presence of alkaloids as detailed below: i. Dragendorff’s Test: To 2 ml of the filtrate was added 1 ml of Dragendorff’s reagent. Formation of orange or reddish brown precipitate indicated the test as positive. ii, Mayer's Test: To 1 ml of test solution or filtrate was added a drop or two of the Mayer's reagent. White or a creamy precipitate confirmed the test as positive. iii, Hager’s Test: To 1 ml of test solution or filtrate, a drop or two of Hager’s reagent formation of yellow precipitate indicated the test as positive. iv. Wagner's Test: Two drops of Wagner's reagent was added to Iml of the test solution, The formation of yellow or brown precipitate confirmed the test as positive for alkaloids. ©) Phytosterols i. Liebermann-Burchard’s Test: The extract (2 mg) was dissolved in 2 ml of acetic anhydride, heated to boiling, cooled and then 1 ml of concentrated sulfuric acid was added. A brown ring formation at the junction and the turning of the upper layer to dark green color confirmed the test for the presence of phytosterols. D) ‘Titerpenoids Salkowski Test: Approximately 2 mg of dry extract was shaken with 1 ml of chloroform and a few drops of concentrated sulfuric acid were added. A red brown color formed at the interface indicated the test as positive for Triterpenoids. AE) Flavonoids i, Shinoda test: A few magnesium turnings and 5 drops of concentrated hydrochloric acid was added drop wise to 1 ml of test solution. A pink, scarlet, crimson red or occasionally green to blue color appeared after few minutes confirmed the test. ii, Alkaline reagent test: Addition of 5 drops of 5% sodium hydroxide to 1 ml of the test solution results in increased intensity of the yellow color which become colorless on addition of a few drops of 2 M hydrochloric acid which indicated the presence of Flavonoids. iii, Lead acetate test: A few drops of 10% lead acetate added to 1m1 of the test solution resulted in the formation of yellow precipitate confirmed the presence of Flavonoids. F) Saponins i, Foam Test: 5 ml of the test solution taken in a test tube was shaken well for five minutes. Formation of stable foam confirmed the test. fi, Olive oil test: Added a few drops of olive oil to 2ml of the test solution and shaken well. The formation of a soluble emulsion confirmed the test. G) Cardiac glycasides i. Keller -Killiani test: Added 0.4 ml of glacial acetic acid and a few drops of 5% ferric chloride solution to a little of dry extract. Further 0.5 ml of concentrated sulfuric acid was added .The formation of blue color in acetic acid layer confirmed the test H) Test for carbohydrates i. Molisch’s test: To 1 ml of test solution added a few drops of 1 % alpha-napthol and 2-3 ml concentrated sulfuric acid. The reddish violet or purple ring formed at the junction of two liquids confirmed the test. ii, Barfoed’s test: 2ml of reagent was added to 2 ml of the test solution, mixed & kept in a boiling water bath for 1 min. Red precipitate formed indicates the presence of monosaccharide. Seliwanoff’s test: To 3 ml of Seliwanoff’s reagent was added to 1 ml of the test sample and heated on a water bath for one minute. The formation of rose red color confirmed carbohydrates. iv. Fehling’s test: Dissolved 2 mg dry extract in 1 ml of distilled water and added Iml of Fehling’s(A+B) solution, shooked and heated on a water bath for 10 minutes. The brick red precipitate formed confirmed the test. 1) Anthraquinone glycosides i. Hydroxyanthraquinone Test: To 1 mi of the extract, added a few drops of 10% potassium hydroxide solution. The Formation of red color confirmed the test. J) Test for proteins i, Biuret test: To 2 ml of the test solution added 5 drops of 1% copper sulphate solution and 2 ml of 10% NaOH .Mix thoroughly. Formation of purple or violet color confirmed proteins. iitK) Test for amino acids llon’s reagent to 1 ml of test solution and heated i. Millon’s test: Added 5 drops of mill on a water bath for 10 min, cooled and added 1% sodium nitrite solution. Appearance of red color confirmed the test. L) Fats and fixed oils Je were added 1 ml of 1% copper sulphate solution and a few To 5 drops of the sampl drops of 10% sodium hydroxide. The formation of a clear blue solution confirmed the test. 3.2.6. ANALYTICAL EVALUATION A) Chromatographic techniques i. TLC-Thin layer chromatography i. HPTLC-High performance thin layer chromatography \ii, -HPLC-High performance/pressure liquid chromatography iy. GLC-Gas chromatography v. CC-column chromatography vi. Gel permeation chromatography vil. Affinity chromatography B) Spectrophptometric methods i. UV- Ultra violet /visible spectroscopy ji, TRnfra Red spectroscopy iii, Fluorescence analysis iv. NMR-nuclear magnetic resonance spectroscopy v. MS-Mass spectroscopy vi, Xeray diffraction vii, RIA-radio immune-assay 3.2.7. BIOLOGICAL EVALUATION «It is employed when the drug cann physical methods. .. inthis method, the response produced by the test drug on a living system is compared with that of the stranded preparation. «Such an activity is represented in units as International units (TU). = Example > Digitalis 11U=76mg of standard > Vit-A 11U-0.344 of standard > Vit/D 11U=0.0250f standard ot be evaluated satisfactorily by chemical and International Units (LU). Dose is termed as| Evalustion of Commercia! Crude Drugs Intended for Use a * SIGNIFICANCE i. The method is generally used when standardization is not done satisfactory by chemical or physical methods. li, When the quantities of the drug /sample are very less, then the drugs are evaluated by biological methods. ili, These methods are performed on living animals, isolating living organ and tissue, animal preparation, and micro-organism (Bioassay). 9 REVIEW QUESTIONS MULTIPLE CHOICE QUESTIONS 1. Palisade ration is the average number of palisade cells beneath ___ epidermal cell, using four continuous epidermal cells for the count. A) One B) Three ©) Five D) None of these 2. Palisade ratio is determined by using ___. A) Simple Microscope B) Camera-Lucida. ©) Both A&B D) None of these 3. Average number of stomata present per __of the epidermis is known as stomatal number. ‘A)Sq.mm B)Sq.cm ©) Sq. pm D) None of these 4. Melting point of colophony is A) 75-85°C B) 45-55 °C ° ©) 65-75°C 1D) 95-105°C 5. Vein-islet Number of Andrographis paniculata is__. A) 17-20 B)9-12 1547 D) None of these 6, Moisture content can be determined by heating the drug at 150°C in an oven to a constant * weight and calculating the loss of weight. A) 150°C B) 105°C. ©) 95°C D) None of these 7. Generally. ethyl alcohol is used for determination of Alcohol soluble extractive. A) 60% B) 80% ©) 95% D) None of these1 ‘ ‘ t ‘ i SHORT ANSWER QUESTIONS 1. Define crude drug? Ans: crude drugs are plant, animal or their parts which after collection are subjected to only drying or making them in to transverse/ longitudinal slices piece or peeling them in some cases, 2. Give the sources of crude drugs. Ans: * Plant origin: whole plant or part of plant like leaves, flowers, seeds and barks. Or vegetable saps, extracts and secretions. * Animal origin: whole animals, glands or organs, extracts and secretions. * Mineral origin: ferrous sulfate, magnesium, zinc, gold etc. 3. Define the term Drug evaluation. ‘Ans: Drug evaluation may be defined as the determination of identity, purity and quality of a drug. 4, Enlist the methods of drug evaluation. Ans: * Organoleptic evaluation * Morphological evaluation * Microscopic evaluation * Physical evaluation * Chemical evaluation * Analytical evaluation * Biological evaluation 5. Define Vein-termination Number. Ans: It is defined as the number of veinlet terminations per. sq. mm of the leaf surface between midrib and margin. °[ Evaluation of Commercial Crude Drugs Intended for Use 6. What do you mean by Refractive index? ‘Ans: When a ray of light passes from one medium to another medium of different density, it is bent from its original path. Thus, the ration of velocity of light in vacuum to its velocity in the substance is said to the Refractive index of the second medium. LONG ANSWERS QUESTION 1. Explain brief about organoleptic evaluation. ‘Ans: Refer Point 32.1. 2... Explain brief about microscopic evaluation. Ans: Refer Point 3.2.3. 3. Explain brief about physical evaluation ‘Ans: Refer Point 32.4. 4. Explain brief about chemical evaluation. ‘Ans: Refer Point 325. VERY LONG ANSWERS QUESTION 1. Explain in detail about Methods of drug evaluation. Ans: Refer Point 3.2.