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Introduction of pharmacognosy & Phytochemistry

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Introduction of pharmacognosy & Phytochemistry

  1. 1. Introduction to Pharmacognosy UNIT-I Mr. Vishal S. Bagul Assistant Professor Department of Pharmacognosy H. R. Patel Institute of Pharmaceutical Education and Research, Shirpur (Maharashtra) Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  2. 2. Content Mr. Vishal S Bagul, Assistant Professor., HRPIPER Definition, History, Scope and Development of Pharmacognosy Source of Drug Organized Drug and unorganized Drug Classification of Drug Quality Control of Drugs of Natural Origin
  3. 3. Definition • The word Pharmacognosy is derived from the Greek "Pharmakon", meaning a drug or poison and “gignosis" meaning to acquire knowledge of and literally meaning “the entire knowledge of drug”. • The scientific study of the structural, physical, chemical and sensory characters of crude drugs of vegetable, animal and mineral origin. • includes also their history, cultivation and collection of the medicinal plants. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  4. 4. History Mr. Vishal S Bagul, Assistant Professor., HRPIPER Hippocrates a Greek scientist, is known as the father of medicine. He worked on human anatomy and Physiology, particularly circulatory system and nervous system. He prepared famous oath for physicians, which is still taken by them. Aristotle and Theophrastus well known philosopher and scientist are known for their writing animal and plant kingdom respectively. • The practice of using herbs to cure disease is as old as human civilization but a drastic change is observed in its practice, as compared to that era. • It is solely possible due to the contributions of many known and unknown scientists. • Plants were used medicinally in India, China, Egypt and reporting or documenting the proof of herbs using effectively.
  5. 5. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Charak complied a group of ten herbs that related to a certain disease. He made 50 such groups which cover almost all the drugs required by physicians for treatment. Then he wrote the book Charak Samhita. Sushruta arranged a large number of drugs into distinct sets depending on their properties. He was one who introduced the practice of surgery. He arranged all his meticulous work into his book Sushruta Samhita
  6. 6. Modern Pharmacognosy • The development of modern Pharmacognosy began during the period of 1930-1960 by the application of a broad spectrum of biological and socio-scientific subjects, including botany, ethno botany, medical anthropology, marine biology, microbiology, herbal medicine, chemistry, biotechnology, phytochemistry, pharmacology, pharmaceutics, clinical pharmacy and pharmacy practice along with modern analytical techniques like paper and thin layer chromatography (TLC), gas chromatography (GC), High performance liquid chromatography (HPLC), Extreme and ultra-pressure liquid chromatography (XLC, UPLC), high pressure thin layer chromatography (HPTLC), Mass spectroscopy, Liquid chromatography combined with mass spectroscopy (LC/ MS), High Resolution Mass Spectroscopy (HRMS). Examples like isolation of penicillin in 1928 by Alexander Fleming from microorganisms and later on commercial production of the same in 1941 by Florey and Chain.
  7. 7. Name of Scientist and their contributions SCIENTIST INVENTION YEAR Friedrich Adam Isolation and Documentation of Mesonic acid from Opium 1805 Karl L Reimann Isolation Nicotine from Tobacco 1828 Albert Niemann Isolation Cocaine from Coca Species 1860 Hardy Isolation Pilocarpine from Pilocarpus 1875 Nagayoshi Nagai Isolation Ephedrine from Ephedra 1885 Kiersten Isolation Podophyllotoxin from Podophyllum 1891 Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  8. 8. • Scope of Pharmacognosy Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  9. 9. • Pharmacognosy is critical in development of different disciplines of science. A pharmacognosist should possess a sound knowledge of the terms used to describe the vegetable and animal drugs as covered under botany and zoology, respectively. The knowledge of plant taxonomy, plant breeding, and plant pathology and plant genetics is helpful in the development of cultivation technology for medicinal and aromatic plants. Plant - chemistry (phytochemistry) has undergone significant development in recent years as a distinct discipline. It is concerned with the enormous variety of substances that are synthesized and accumulated by plants and the structural elucidation of these substances. Extraction, isolation, purification and characterization of phytochemicals from natural sources are important for advancement of medicine system. • Many of drugs today are being derived from herbs molecules. • Many new molecules every year are being introduced through this field alone, so it is great source of lead molecules. • Marine sources possess about 5 Lakh species with lots of possibilities of developing medical purpose. • Source of natural drug improves the importance of plants and animals. • Beneficial against allopathic medicine because of less chances of toxicity issue to human being • These are less cost effective formulations. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  10. 10. Development of Pharmacognosy • Modern Pharmacognosy occurred during 1934 - 1960. • This development was mainly due to some events like • Discovery of Penicillin in 1928 by Alexander Fleming • Isolation of Reserpine 1952 • Anticancer properties of Vinca rosea • Discovery of Dioscorea was utilized to prepare semisynthetic • Steroidal hormones. • Pharmacognosy as an applied science. • It played a crucial role in the development of different disciplines of science. • Some isolated components from plants were made into different formulations and used as drugs Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  11. 11. Applications of Pharmacognosy Plant bioactive Extraction and Isolation • This type of approach is useful for the investigation of alkaloids, antioxidants, bioflavonoids, terpenoids and other compounds as promising therapeutics from an immense pool of plant biodiversity. Pharmacognosy in drug development • Pharmacognosy is used by pharmaceutical companies to screen, characterize and produce new drugs for the treatment of human disease. • naturally occurring drugs cannot be mass-produced, so they must be studied in order to develop synthetic biosimilars. Development of nano fertilizers and nano medicines: • Nanotechnology in drug delivery is exemplified by nanocrystals, liposomes, nanoparticle-protein conjugates, magnetic nanoparticles, nanogels and biodegradable nanoparticles. Fertilizer particles can be coated with nano membranes that facilitate slow and steady release of nutrients thereby reducing loss of nutrients and enhancing its use efficiency of medicinal plants
  12. 12. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  13. 13. • PLANT SOURCE • Plant have always been a rich source of lead compounds (e.g. Alkaloids, Morphine, digitalis, Quinine and nicotine). • Almost all parts of the plants are used i.e. leaves, stem, bark, fruits and roots. 1. Leaves: • The leaves of Digitalis Purpurea are the source of Digitoxin and Digoxin, which are cardiac glycosides. • Leaves of Eucalyptus give oil of Eucalyptus, which is important component of cold & cough syrup. • Tobacco leaves give nicotine. • Atropa belladonna gives atropine. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  14. 14. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Flowers: Poppy Papaver somniferum gives morphine (opoid) Rose gives rose water used as tonic. Fruits: Senna pod gives anthracine, which is a purgative. fennel is a source of fenacone and it having carminative action. Seeds: Seeds of Nux Vomica give strychnine, which is a CNS stimulant. Castor seeds give castor oil. Roots: Ipecacuanha root gives Emetine, used to induce vomiting as in accidental poisoning. It also has amoebicidal properties. Bark: Cinchona bark gives quinine and quinidine, which are antimalarial drugs. Quinidine also has anti-arrythmic properties. Stem: Ephedra stem contain ephedrine it use for appetite suppressant
  15. 15. ANIMAL SOURCE • Pancreas is a source of Insulin, used in treatment of Diabetes. • Sheep thyroid is a source of thyroxin, used in hypertension. • Cod liver is used as a source of vitamin A and D. • Blood of animals is used in preparation of vaccines. • Stomach tissue contains pepsin and trypsin, which are digestive juices used in treatment of peptic diseases in the past. MARINE SOURCES • The drug obtained from marine organisms which are being conventionally used like shark and cod-liver oils, sodium alginate, agar-agar, chitin etc. • Over 70% of the earth's surface is covered by oceans which contain 95% of the earth's biosphere.
  16. 16. TISSUE CULTURE Is the growth of tissues or cells separate from the organism. This is typically facilitated via use of a liquid, semi-solid, or solid growth medium, such as broth or agar. Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants. The plant tissue culture technique is important because isolation of bioactive compounds from the medium is very easy, rare and endangered plant species are micro propagated and cultivated in mass scale, production of immobilized plant cell for future use and even biochemical conversion is easy.
  17. 17. • Organized drugs consist of the cellular organization in the form of anatomical features. These are mostly the crude drugs from plant sources. • Microscopical and anatomical studies are preeminent for such crude drugs. These can be used directly in medicine or can be used by modifying or by extracting the active ingredient from it. Examples • Bark: Cascara, Cassia, Cinchona • Fruit: Anise, Capsicum, Caraway • Seeds: Black Mustard, Cardamom, Nux-vomica • Roots and Rhizomes: Aconite, Garlic, Ginger, Gingsing Mr. Vishal S Bagul, Assistant Professor., HRPIPER • The unorganized drugs do not have the morphological or anatomical organization as such. These are the products which come directly in the market but their ultimate source remains the plants, animals or minerals. • These products may be solid, semisolid or liquid and the physical, chemical and analytical standards may be applied for testing their quality and purity. Examples • Dried Extract: Agar, Black catachu, Pale catachu, Pectin • Waxes: Beeswax, Spermaceti • Gums: Acacia, Gaur gum • Volatile oil: Coriander, peppermint, Sandalwood ORGANIZED DRUG UNORGANIZED DRUG
  18. 18. CLASSIFICATION OFDRUGS Alphabetical classification Morphological classification Pharmacological classification Chemical classification Taxonomical classification Chemotaxonomic classification Sero-taxonomical classification
  19. 19. Alphabetical classification • Crude drugs are arranged in alphabetical order of their Latin and English names (common names) or sometimes local language names (vernacular names). • Some of the pharmacopoeias, dictionaries and reference books which classify crude drugs according to this system are as follows: 1. Indian Pharmacopoeia 2. British Pharmacopoeia 3. British Herbal Pharmacopoeia 4. United States Pharmacopoeia and National Formulary 5. British Pharmaceutical Codex • In European Pharmacopoeia these are arranged according to their names in Latin where in United States Pharmacopoeia (U.S.P.) and British Pharmaceutical Codex (B.P.C.), these are arranged in English. Example: Acacia, Bees wax, Castor oil, Datura …….etc Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  20. 20. Advantages • It is very simple to use and operate. • It is very easy to find out the specific data and go to a specific entry. • There is no repetition of single entry. Disadvantages • It does not give any idea regarding chemical constituents, uses and other information regarding plants.
  21. 21. Morphological classification • Plants are made up of different parts. Different plants can be grouped according to the part which is being utilized . This system is truly useful, as it gives guidance regarding the harvesting of plants. E.g. Leaves: Vasaka, Senna, Digitalis Bark : Cinchona, Arjuna, Cinnamon Roots : Rauwolfia, Ashwagandha Fruits : Coriander, Fennel Whole plant: Tulsi, Vinca Latex : Opium
  22. 22. Pharmacological classification: In this system of classification, the plants are arranged accordance with their pharmacological or biological activity. Different plants give certain medicinal effects E.g. Anti-diabetic: Fenugreek, Neem Anti-cancer: Taxus, Vinca Anti-malarial: Cinchona, Artemisia Advantages: It prepare a list of substitutes which can be utilized for treating same condition. Disadvantages : 1.Some plant possess more than one activity; in such cases the plant name is repeated. 2 .Pharmacological classification is unable to give data regarding parts of plants being used especially in case of unorganized drug, it create problems.
  23. 23. Chemical classification: • Depending upon the active constituents, the crude drugs are classified. The plants contain various constituents in them like alkaloids, glycosides, tannins, carbohydrates, saponins, etc. Irrespective of the morphological or taxonomical characters, the drugs with similar chemical constituents are grouped into the same group. • Therapeutics effects observed in plants, only depends on the chemical constituents. The presence of one of the chemical constituents can be correlated to certain specific biological effects.
  24. 24. • Advantages: a. It arranges the plant according to their chemicals, which after flexibility to study further subtypes. b. For studying the plants, this is the best method. c. Due to similar chemical classes many times similar extraction and isolation procedure can be employed. E.g. Stas and Otto • Disadvantages It does not resemble to any pharmacological benefits of plants. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  25. 25. Taxonomical classification This is also referred to as Biological classification. It is an old system initiated by Linax. In this method plants are mentioned with various ranked levels. The levels contain plants with some similar morphological properties. This approach of classification allows us to understand various correlations among the various plant species. Kingdom: Plantae-It contains all sort of plants, green algae, ferns etc. Division: Angiospermae- Flowering plants Class: Dicotyledons- Seeds can be divided in two equal parts. Order: Rosales- Flower producing order Family: Leguminosae- Nitrogen fixing bacteria in roots Genus : Trigonella- Leaves appear in groups of three Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  26. 26. Mr. Vishal S Bagul, Assistant Professor., HRPIPER 1. We can study the different drugs with same family at a time due to its resemble characteristics of that particular family. 2. Detail botanical information and evolutionary development of plant can be studied. Advantages Disadvantages 1. It fails to recognise organised and unorganised drugs………………..…………. 2. Many drugs are not entire plants and represent part of plant that have been processed systematically that are not classified by this system 3. It fails to give the data of chemical nature of active constituent and therapeutic significance of crude drug.
  27. 27. Chemotaxonomic classification: • This system of classification on the chemical similarity of a taxon, i.e. it is based on the existence of relationship between constituents in various plants. • There are certain types of chemical constituents that characterize certain classes of plants. This gives birth to entirely a new concept of chemotaxonomy that utilizes chemical facts/characters for understanding the taxonomical status, relationships and the evolution of the plants. • For example, tropane alkaloids generally occur among the members of Solanaceae, thereby, serving as a chemotaxonomic marker. Similarly, other secondary plant metabolites can serve as the basis of classification of crude drugs.
  28. 28. Advantages • It gives more scope for understanding the relationship between chemical constituents, • their biosynthesis and their possible action. • The characters most often studied are secondary metabolites of pharmaceutical • significance such as alkaloids, glycosides, flavonoids, DNA hybridization, amino acid • sequencing in proteins etc. • It provides degree of hybridization and breeding analysis Disadvantages: • This system is fails to identify the organized and unorganized crude drugs in their morphological studies. • This system fails to understand the therapeutic nature of the crude drugs. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  29. 29. Serotaxonomical classification The study of antigen-antibody reaction is called serology. The application of serology in solving taxonomic problems is called Serotaxonomy. The technique is based on the highly specific relationship between antigens and antibodies produced in response to them. It expresses the similarities and the dissimilarities among different taxa, and these data are helpful in taxonomy. It determines the degree of similarity between species, genera, family, etc., by comparing the reaction with antigens from various plant taxa with antibodies present against a given taxon.
  30. 30. Gum and Mucilage Gum • is a sap or other resinous material associated with certain species of the plant kingdom. This material is often polysaccharide-based and is most frequently associated with woody plants, particularly under the bark or as a seed coating. The polysaccharide material is typically of high molecular weight and most often highly hydrophilic or hydro colloidal. Mucilage • a polysaccharide substance extracted as a viscous or gelatinous solution from plant roots, seeds, etc., and used in medicines and adhesives. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  31. 31. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  32. 32. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Dried Latex • is a stable dispersion (emulsion) of polymer micro particles in an aqueous medium. Resins • Resin is a solid or highly viscous substance of plant or synthetic origin that is typically convertible into polymers. Oleo resin- • A natural or artificial mixture of essential oils and a resin, e.g. balsam. Oleo gum resin- • a solid plant exudation (as asafetida) consisting of a mixture of volatile oil, gum, and resin
  33. 33. Quality control of Drugs of Natural Origin • Adulteration of drugs of natural origin. • Evaluation by organoleptic, microscopic, physical, chemical and biological methods and properties. • Quantitative microscopy of crude drugs including lycopodium spore method, leaf constants, • Camera lucida and diagrams of microscopic objects to scale with camera lucida Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  34. 34. Quality Control Mr. Vishal S Bagul, Assistant Professor., HRPIPER As per Pharmacopeial definition, the quality control of herbal drugs are based on three terms likely Identity, Purity and Assay. Identity 2. Purity 3. Assay Quality of herbal drugs are defined as the status of a drug that is determined by its evaluation in terms of identity, purity, content and other physical, chemical and biological properties and manufacturing processes for formulated drugs.
  35. 35. Adulteration • It is a practice of substituting original crude drug partially or wholly with similar looking substances. Reason For Adulteration • To get more profit. • Due to shortage of drugs. • In case of costlier drug. • For banned drugs Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  36. 36. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  37. 37. Mr. Vishal S Bagul, Assistant Professor., HRPIPER 1. Substitution with artificially manufactured substances Such a type of adulteration is found in case of costlier drugs. Some plants or parts of plants rarely available because of the season or climatic conditions. Some artificial substances with similar looks are available at cheaper rate. Example: Artificial invert sugar for Honey 2. Uncertain identity of the drug Sometimes due to the confusion of the authenticity of the plant, drug identity becomes uncertain. Like Chichona bark it has different species like Cinchona calisaya, C. officinalis, C. ledgeriana etc. are considered.
  38. 38. 3 Seasonal availability of drugs: Some drugs are available in a specific season so other drugs can be introduced in their absence which has same action. For example, Trianthema portulacastrum can be used in seasonal absence of Boerhavia diffusa 4 Harmful adulterants: This is happened with unorganized and liquid drugs. Wastes materials are collected and mixed with original drugs. Eg. Pieces of amber colour glass with colophony. 5. Substitution with totally different drugs: This practice is generally done in case of oil. Like cotton seed oil in place of olive oil. Sometimes barks also substitute original bark. 6. Substitution of species in the same family: This practice is done in case of dried leaves, roots or stems. Example: Dog senna in place of Indian seena, but both have same family (Leguminosae)
  39. 39. 7. Substitution with exhausted drugs- Volatile oil containing drugs are generally adulterated by method same drug are being used as an adulterants but they do not contain medicinally active constituents as they are already extracted out. Some time additives are added to impart colour as well as taste. Original tea powder and filtered tea powder after use look similar but chemical constituent from the filtered tea powder have been extracted and missing Eg – Fennel, Coriander etc Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  40. 40. Quality Control a system of maintaining standards in manufactured products by testing a sample of the output against the specification. Evaluation Confirmation of its identity and determination of its Quality and Purity and Detection of nature of adulterant by various parameters like Morphological, Microscopical, Physical, Chemical and Biological observations
  41. 41. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  42. 42. MORPHOLOGICAL / ORGANOLEPTIC • Organoleptic evaluation of crude drugs refers to the evaluation of a drug by colour, odour, taste, size and shape, occasionally the sound or snap of fracture and special fetures including touch, texture, etc.. • It is a technique of qualitative evaluation based on study of morphological and sensory profiles of whole drugs. Advantage: It provides simplest and quickest means to establish the identity & purity and thereby ensure quality of particular sample Disadvantage: Judgement may vary from person to person and time to time. Example: • Fractures of barks like cinchona, cascara. • Sweet taste of liquorice
  43. 43. BOTANICAL MACROSCOPIC • Examine the untreated sample. If necessary, a magnifying lens (6x to 10x) may be used. Wetting with water or reagents, as required, may be necessary to observed the characteristics of a cut surface. • The texture is best examined by taking a small quantity of material and rubbing it between the thumb and forefinger, it is usually described as ‘smooth’, ‘rough’ a) Shape and size b) Colour and external marking c) Fracture and internal colour d) Odour and taste
  44. 44. MICROSCOPICAL • Leaf constants or diagnostic characters of leaf- • Stomatal numbers- • Is average numbers of stomata per.sq.mm of epidermis of leaf. • Stomatal Index- • Is the percentage which number of stomata form to the total number of epidermal cells, each stoma being counted as one cell. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Epidermal cell
  45. 45. Types of stomata
  46. 46. Vein –Islet number- • It is defined as number of vein-islet present per square mm area of leaf surface Vein termination number- • It is number of vein terminating at particular end per square mm area of leaf surface Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  47. 47. Calcium Oxalate Crystal: • Calcium oxalate crystals are present in almost all parts of the plant. The Calcium oxalate crystals are either the monohydrate whewellite form or the dihydrate weddellite form. They are identified by X-ray diffraction; Raman microprobe analysis and infrared spectroscopy are the most accurate. Under microscope they are visible as colorless when treated with chloral hydrate solution. Functions: • They give protection to the plant against environmental stress. • They help in identification of crude drugs. • They help in detection of adulterants. • They help in identification of plants in same family.
  48. 48. Trichomes • Trichomes is the term used to refer to tiny outgrowths from the plant epidermis. Although the term "trichomes" generally refers to outgrowths ranging from small hairs to larger outgrowths like thorns, it is typically used to refer to the tiny hairs that can be seen emerging from the surfaces of leaves and other epidermal surfaces of plants. Trichomes are classified in main three types on the basis of structure and number cells present 1) Covering or non-globular or clothing trichomes- eg., Nux-vomica, Tea, Cannabis. 2) Glandular trichomes- eg., Vasaka, Piper, Digitalis. 3) Hydathodes or special type of trichomes- eg., Piper betal, London pride. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Functions of Trichomes: 1. Plant defense against insects 2. Generally a dense covering of woolly trichomes controls the rate of transpiration. 3. They also reduce the heating effect of sunlight. 4. They aid in the protection of plant body from outer injurious agencies. 5. Chemicals produced in the glandular tip can deter feeding or the trichome can physically prevent the insect from reaching and feeding on the leaf.
  49. 49. PHYSICAL Qualitative 1. Refractive Index- • When a ray of light passes from one medium to another of different density, it is bent from original path. Thus, the ratio of the velocity of light in vacuum to its velocity in the substance is termed as refractive index of the second medium. Depending upon purity it is constant for a liquid. Eg- Castor oil 1.4758-1.4798 Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  50. 50. 2. Viscosity: • Viscosity is a measure of the resistance of a fluid which is being deformed by either shear or tensile stress. The most common method of determining kinematic viscosity in the lab utilizes the capillary tube viscometer. Examples: Clove oil: 0° to – 1.5°, Cinnamon oil: 0° to – 2°. 3. Foreign Organic Matters: • Anything extra present in the drug which is not complying with the authentic drug may be considered as a foreign matter. The foreign matter can be present in the drug due to improper harvesting. The source of foreign organic matter can be animal excreta, insect or mould and is determined by sedimentation or floatation method. • It determines presence of organic as well as inorganic foreign matter. • It gives an idea weather drug is pure or adulterated Examples: Curcumin – not more than 2.0%; Neem – not more than 1.5% Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  51. 51. Mr. Vishal S Bagul, Assistant Professor., HRPIPER 4. Solubility- • The presence of adulterants can be determined by solubility studies. • Examples: Castor oil is soluble in 3 volumes of 90 percent alcohol, while the adulterated form may shows good solubility in alcohol. 5. Extractive Values: • All the chemical constituents are soluble either in polar, semi polar or organic solvents. Total soluble constituents of the drug in any particular solvent or mixture of solvents may be called its extractive value or percent extractive. Significances of Extractive Values: • This method is important when the constituents of drugs can’t be readily estimated by any other means. • It indicates the nature of chemical constituents present in drugs. • It helps in identification of adulterants.
  52. 52. 6. Ash value: Ash value is useful in determining authenticity and purity of sample and also these values are important qualitative standards. • Acid insoluble ash value indicates siliceous impurities. • Water soluble ash value gives an estimation of inorganic contents Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  53. 53. 7. Moisture content Moisture content refers to the number of water molecules that become incorporated into a Crude and food product. 8. Swelling index The swelling index is the volume in ml taken up by the swelling of 1g of plant material under specified condition. Its determination is based on the addition of water or a swelling agent as specified in the test procedure for each individual plant material. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  54. 54. • Plant materials containing saponins cause persistent foam when the decoction is shaken. Foaming ability of plant material and their extracts is measured by foaming index Mr. Vishal S Bagul, Assistant Professor., HRPIPER 9. Foaming Index:
  55. 55. CHEMICAL • It is qualitative as well as quantitative type of evaluation. • Chemical tests and chemical assays are prime important. • Isolation, purification, and identification of active constituents . • Preliminary phytochemical screening is qualitative evaluation. • Qualitative chemical tests are useful in detection of adulteration. • eg., Copper acetate is used for detection of balsams and waxes. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Qualitative
  56. 56. • Test for alkaloid Dragendoff’s test Mayers test Hagars test Wagnar test • Test for carbohydrates Molisch test Barfoed’s test
  57. 57. QUANTITATIVE • Colorimetric Method: It is a method of determining the concentration of a chemical element or chemical compound in a solution with the aid of a color reagent. It is applicable to both organic and inorganic compounds and may be used with or without an enzymatic stage. The method is widely used in medical laboratories and for industrial purposes, e.g., the analysis of water samples in connection with industrial water treatment. • UV-Visible Spectroscopy
  58. 58. • Volumetric Method: It is a quantitative analysis of liquids or solutions by comparing the volumes that react with known volumes of standard reagents, usually by titration. A reagent is prepared as a standard solution, acts as titrator. A known concentration and volume of titrant reacts with a solution of analyte or titrand to determine concentration. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  59. 59. • Photometric Method: It is the set of methods of quantitative chemical analysis based on the relationship between the concentration of a substance in a solution or gas and the absorption of radiation. In this method the intensities of the monochromatic components of transmitted radiation are scanned Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  60. 60. • Gravimetric Method: It is the quantitative determination of a substance by the precipitation method of gravimetric analysis involving isolation of an ion in solution by precipitation reaction, filtering, washing the precipitate, conversion of precipitate to a product of known composition, and finally weighing the precipitate and determining its mass by difference. There are four fundamental types of gravimetric analysis: physical gravimetry, thermogravimetry, precipitative gravimetric analysis, and electrodeposition. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  61. 61. BIOLOGICAL • Measurement of biological effect and Pharmacological activity of new or chemically undefined substance. • Determination of the side effect profile, including the degree of the drug toxicity. • Measurement of the concentration of know substance Cardiac glycoside are evaluated by this method using frogs, cats, pigeons
  62. 62. Microbiological assay • The method of measuring compounds such as vitamins and amino acids using microorganism. • As per IP, two methods are used for the assay • Cylinder plate method or Cup Plate methods Cylinder plate method: based upon the diffusion of the antibiotic throughout solid culture media. Cup Plate methods: antibiotic containing cylinder is diffused into the agar layer containing the microorganisms
  63. 63. Analytical Methods • In this method various chromatographic technique as well as spectroscopic methods are used for the sample. Thin Layer chromatography (TLC): used for separation of Mixture with the principal of adsorption. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  64. 64. High Performance Thin Layer Chromatography (HPTLC) • This system is a versatile modern analytical technique with reference to its excellent automation, optimization, multidimensional application. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  65. 65. High performance Liquid chromatography (HPLC) • It is highly improved column chromatography. By this methods solvent allowed passing though the stationary column chromatography. Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  66. 66. Gas Chromatography (GC) • It is used for separation and analysis of vaporized compound which are non decomposable. • Nitrogen gas are mostly used for this chromatography Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  67. 67. UV-Visible Spectroscopy • This method refer to absorption spectroscopy in the visible spectral region. • UV range id 800-200nm • Morphine is determined at 286nm Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  68. 68. Infra-red Spectroscopy (IR) • It is used for identification of functional group and structure elucidation, identification of substance, detection of impurities. • (14000-4000 cm-1) Mr. Vishal S Bagul, Assistant Professor., HRPIPER
  69. 69. Lycopodium spore method • This method is used to identify the crude drugs when the chemical and physical methods are inapplicable. • This method is also useful to detect the adulteration present in the crude drugs containing starch grains. • Lycopodium spore method was performed on ingredients of Shatavar churna, an ayurvedic formulation used as immunomodulator • Examples: Adulterated drug containing starch can be determined by counting the number of starch grains per mg and calculating the amount from the known number of starch grains per mg of the pure starch. The percentage purity of an authentic powdered ginger is calculated using the following equation:
  70. 70. Mr. Vishal S Bagul, Assistant Professor., HRPIPER Significance: • Determination of foreign organic matter. • Determination of percentage purity of drugs. • Detection of adulterant
  71. 71. Leaf constants • Important quantitative Microscopic evaluations which can be used to identify and distinguish between some closely related species. • Stomata • Stomatal numbers • Stomatal Index • Vein –Islet number • Vein termination number- • Calcium Oxalate Crystal • Trichomes • Palisade ratio
  72. 72. CAMERA LUCIDA • It is an optical device or instrument in which rays of light are reflected by a prism to produce an image on a sheet of paper, from which a drawing is made. It works on simple optical principle reflecting beam of light through a prism and a plane mirror.
  73. 73. • Camera Lucida, when attached with a compound microscope, helps drawing microscope images of objects on paper. • It works on simple optical principle reflecting beam of light through a prism and a plane mirror. The microscopic image of the object is reflected by the prism on to the plane mirror and there from the image is reflected on to the plane paper. • The observer moves the pencil on the lines of the image and draws a correct and faithful figure of the object on the paper. There are three main parts of a camera lucida the attachment ring, the prism, and the mirror. • The attachment ring attaches the camera lucida with the body tube of the microscope. The prism rests just above the eyepiece when the instrument is attached with the microscope.
  74. 74. Thank You

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