STANDARDIZATION AND QUALITY CONTROL OF HERBAL DRUGS.pptx

METHODOLOGY AND QUALITY CONTROL, STANDARDIZATION OF HERBAL DRUGS- PHARMACOGNOSY AND PHYTOCHEMISTRY

1. STANDARDIZATION AND QUALITY CONTROL OF HERBAL DRUGS

2. Introduction of Herbal drugs Herbal products have been used since long times as medicine/drug for the treatment of a numerous diseases. The basic resources of medicine come from nature and they are used as medicaments since from ancient time till today. Each plants is like bio-factory which is capable of synthesizing infinite number of highly complex and unusual chemical substances which posses pharmacological or other excipients aid which is called as metabolites. There are at least more than 120 different chemical substances originated from plants that are considered as important drugs currently in use in the world. WHO has delivered some terms related to herbal drugs, according to their definitions. Herbal medicines include herbs, herbal materials, herbal preparations and finished herbal products.

3. During the ancient time traditional medicine as include diverse health practices, knowledge and beliefs, spiritual therapies, manual technique and exercises practice are applicable singularly or in combination to maintain well being as well as diagnosis or prevent illness. As according to ancient civilization and development this herbal drugs are utilize for health care applied in indigenous system of medicine like in – Ayuveda , Siddha and Unani , Naturopathy, C hinese med system etc. Herbal drugs which are use in cure of illness are utilize or given in crude or raw state, which is pure in nature and directly intake without formulating or in some times formulated in simple dosage form. The un processed herbal drugs which are obtain from the source lower or higher plants are known as crude drugs and it is divided on the basis of nature and appearance.

4. Types of crude d rug’s Organized crude drugs Unorganized crude drugs It consist of a direct parts of the plant which are made of cell or definite structure. like leaves, barks wood, root, rhizome, seed, fruits, flowers, stem, hair and fibers etc They are appearances in solid nature. Botanical or zoological terminology can be used to describe these drugs. Microscopic characteristic are one of the important criteria for identification. Digitalis leaf, cinchona bark, clove etc. It consist of derived product obtain from plants and animal by process of extraction and purification. Like latex, juice, gum, resins, oil and fat. These are solid, semisolid or liquid in nature. Botanical or zoological terminology are also used for it but physical character are mainly applicable like Solubility, density, optical rotation, refractive index etc. Chemical test and physical standards are confirmatory test for identification. Aloe- vera flesh , agar, opium, castor oil, bees wax, enzymes etc.

5. Cultivation Drying Natural (sun drying) Artificial (Oven, tray drying, vacuum drying and spray drying) Harvesting Storage and Packaging Market sale and dispensing Garbling (dressing) Cultivation, collection and preparation of herbal crude drugs Sale to herbal preparation Industry

6. What are h erbal drugs ? The term “herbal drugs” are denotes that plants and its parts that have been converted into phyto -pharmaceutical, for therapeutic role by means of simple processes involving harvesting, drying and storage.

7. Definition of herbal drug according to WHO Are – A finished and labelled products that contains an active ingredients which are responsible for therapeutic features and it consist of aerial or underground parts of plant or other plant material, in some time it contains combination, whether in crude state or a formulation. Are not – Medicines that containing the combination of plant material along with chemically defined active substance. (As defined by World Health Organization)

8. Classification of h erbal Drug’s Classification of the herbal drugs are based on the formulation and its dosage state that are :- Phytomedicines or phyto pharmaceuticals sold as over the counter (OTC) product in modern dosage forms such as tablets, capsules, liquids for oral use etc. Dietary supplements containing herbal products, also called as Neutraceutical available in modern dosage forms which is use for health and fitness. Herbal medicines consisting of either crude, semi processed or processed medicinal product. Ayurvedic formulation and dosage form. More over herbal drugs are also divided on the basis of nature and number of drug composition contain within a formulation.

9. HERBAL DRUGS Mono herbal Formulation (Single or crude drugs) Poly herbal Formulation (multi herbal drugs) TYPES OF HERBAL DRUGS Mono herbal formulation or (Single or crude drugs ) – A formulation which consist of only single drug which is called as main active ingredient which posses therapeutic feature. It is simple called as crude drug but some time it is formulated in dosage form as per need. Poly herbal formulation or (multi herbal drugs) – A formulation which consist of multiple number of herbs drugs and have different pharmacological feature as according to it requirement for particular treatment.

10. TYPES OF HERBAL DRUGS Mainly whole, fragmented or cut plants, parts of plant in dried form, some times is taken fresh. It also include algae, fungi, lichens other microbes. Like – S pirulina . It can be obtain by simple process like decoction, maceration. Features of Mono herbal Formulation (Single or crude drugs )

11. TYPES OF HERBAL DRUGS Features of Poly herbal Formulation (Multi herbs drugs ) It consist of different medicinal plants and natural excipients, which have multiple constituents. It ability to posses a pharmacological response as per need along with multiple function. Poly herbal Formulation anti dandruff herbal Shampoo

12. What is Quality control ? Quality control (QC) is a procedure or set of procedures which evaluate to ensure that a manufactured or a finished product, performed services adheres to defined set of quality criteria. Quality control (QC) is defined as a procedure or set of procedure or activities intended to ensure that a quality in products. The activities focus on identifying the defect in the actual products by specific evaluation parameter’s.

13. Identify the defect Safety Efficacy Purity Content Good Laboratory Practices Good Manufacturing Practices Standard as following Factor’s Effecting Quality Control

14. Role of q uality control in h erbal drugs The role of quality control in herbal drugs is to know the efficacy and safety of herbal products. It can be define as status of a drug that is determined by identity, purity, content, and other chemical, physical or biological properties or by the manufacturing processes. It is use to find out the qualitative estimation of adulterate and substitute with in product. Quality control is a term that refers to process which involve in maintaining the quality and validity of manufactured product. In general all the medicines whether they are of synthetic or plant origin, should fulfill the basic requirements of being efficacious and safe, and this can be archived by suitable clinical trials.

15. What is standardization ? Standardization means “ Standardization is a framework of agreements to which all relevant parties in an industry or organization must adhere to ensure that all processes associated with the creation of a good or performance of a service are performed within set guidelines .” Standardization of herbal drugs refers to “Confirmation of its identity and determination of its quality, purity and detection of nature of adulterants by various parameter like- morphological, microscopical , Physical, chemical and biological observation.

16. Standardization and quality evaluation of herbal drugs Standardization of herbal drugs means to confirmation of its identity, quality and purity. Standardization of herbal formulations is essential in order to assess of quality drugs, based on the concentration of their active principle, physical, chemical, physcio -chemical standardization and in vitro, in-vivo parameters. It is necessary to maintain reproducible efficacy and safety of phyto -pharmaceutical therefore, if phyto -pharmaceutical have to regards as rational drug should be standardized and pharmaceutical quality must be approved. WHO (World Health Organization), ICH (International Council of Harmonization) and EU (European union) these are research g uidelines for Evaluating the Safety and Efficacy of Herbal Medicines, which design a protocol and standard guidelines for evaluation studies.

17. Classification of herbal drug s tandardization

18. Standardization and Quality control o f Herbal drugs Botanical Method Physical Method Chemical Method Instrumental Method Biological Method Macroscopical (Morphological studies) Microscopical (Histological studies) The qualitative chemical tests are useful in identification of chemical constituents and detection of adulteration. Moisture contain Ash value Extractive value Viscosity Density Solubility Bitterness value Swelling index Foaming index Specific gravity Spectroscopical analysis Chromatography analysis I solation organs of living animal Animal modal studies Microbial modal

19. Macroscopic method evaluation It initial examination of the drugs which can be used to identify the physical appearance of organized and unorganized drugs by sensory characters. Its refer to evaluation of drugs by colour , odour , taste, shape and size, special characteristic like touch, texture, fracture, sound, aroma etc . It also classify the organized or unorganized nature of herbal drug This method is very common which is also called as morphological evaluation . It is use to evaluate the adulteration and substitution of crude drug by sensory examination. It also give the information regarding the agriculture practices of crude drug and factor which influences the cultivation and collection. Eg . Wood fractured surfaces in cinchona, quillaia and cascara barks and quassia Aromatic odour of umbelliferous fruits and sweet taste of liquorice . The wavy shape of rauwolfia , pungent taste of capsicum and ginger, brown colour of cinnamon, odour and taste of spice-drugs like, asafoetida , black pepper, nutmeg, caraway, cumin etc. are important diagnostic organoleptic characteristics.

20. Macroscopic method evaluation Asafoetida ( Ferula foetida ) Synonyms – Asafoetida , devil drug Bio source – olea -gum resin obtain by making incision in rhizome or root of ferula foetida Family - Umbelliferae Organoleptic (Morphological) feature :- Morphological feature Description Colour Yellowish –white changing to reddish brown Odour Aromatic and persistent Taste Bitter and acidic Shape Tears are 0.5 to 3 cm Size Lumpy mass, round and irregular

21. Microscopical method evaluation It involves detailed examination of the drugs and it can be used to identify the organized drugs by their known histological characters . It is mostly used for qualitative evaluation of organized crude drugs in entire which help for detecting various cellular tissues like trichome’s , stomata, starch granules, calcium oxalate crystals and aleuronic grains etc. Starch and hemicelluloses is identified by blue color with iodine solution , lignified tissues give pink strain with phloroglucinol and HCl etc . mucilage is stained pink with ruthenium red can be used to distinguish cellular structure. Quantitative aspects of microscopy includes study of stomata number and index, palisade ratio, vein islet number, size of starch grains, length of fibers etc. W hich plays a very important role in the identification of drug.

22. Microscopical method evaluation T ransverse section of datura leaf  Transverse section of Fennel fruit Longitudinal of Cinnamon bark

23. Microscopical method evaluation Microscopical method also consist of microscopic linear measurement and quantitative microscopy are also covered under this technique of evaluation. The following microscopical measurement are :- Stomatal number – It is the average number of stomata present in per square mm of the epidermis. Stomatal index- It is a percentage which the number of stomata form to the total no. of epidermal cells, each stoma counted as one cell. It can be calculated by formula. I = S x 100 (E+S) I – stomatal index, S – No. of stoma per unit area, E - epidemal cells in same area c. Vein islet number- The no. of vein islet present in per sq mm of leaf surface. d. Palisade ratio- The Avg no. of palisade cells, present on one epidermal cell.

24. Microscopical method evaluation Stomatal number:- Datura stramonium – 87 stomatal no. Hyoscyamus niger – 125 stomatal no. Stomatal index:- Atropa belladonna – 20.2 to 23.0 Indian senna – 17.0 to 20.2 Vein islet number:- Digitalis purpurea – 02 to 5.5 Cassia angustifolia – 19 to 23 Palisade ratio:- Atropa belladonna – 06 to 10 Digitalis purpurea – 3.7 to 4.5

25. Chemical method e valuation e. Identification test for tannin:- - ferric chloride test - Gold beater skin test - Gelatin sol and sodium chloride solution test Identification test for Carbohydrate:- - Molish reagent test – α napthnol solution test - Fehling solution test

26. Chemical method e valuation Most of drugs have definite chemical constituents to which their biological or pharmacological activity is attributed. Qualitative chemical test are used to identify certain drug or to test their purity. Isolation , purification, identification of active constituents is based on chemical methods of evaluation. Evaluation test of resins : acid value, sulphated ash Evaluation test of balsams: acid value, saponification value, bester values. Evaluation test of volatile oils : acetyl and ester values This covers screening, isolation, identification and purification of the chemical components. Chemical analysis of the drug is done to assess the potency of vegetable material in terms of its active principles. The chemical screening or tests may include colour reaction test, which help to determine the identity of the drug substance and possible adulteration.

27. Chemical method e valuation Identification test for alkaloids:- - Mayer’s reagent (Potassium mercuric iodide sol) - Dragendroff’s reagent (Potassium bismuth iodide sol) - Wagner’s reagent (iodine and potassium iodide sol) - Hager’s reagent (Sat sol of picric acid) Identification test for glycoside:- - Saponin test - Cyanophoric test - keller Killiani test Identification test for Volatile oil:- - Alcoholic sol of Sudan III dye test - Alkana tincture test d. Identification test for fixed oil and fats:- - Sodium hydroxide test - Sodium hydrogen sulphate test - Alcoholic solubility test

28. Active constituents which are present in herb or herbal product

29. Metabolites of plants

30. Physical method evaluation Physical constants are sometimes taken into consideration to evaluate certain drugs. These include moisture content, specific gravity, optical rotation, refractive, melting point, viscosity and solubility in different solvents. All these physical properties are useful in identification and detecting of physical nature of constituents present in plants. Moisture content- The moisture content of a drug will be responsible for decomposition of crude drugs either producing chemical change or microbial growth. So, the moisture content of a drug should be determined and controlled. The moisture content is determined by heating a drug at 105o c in an oven to a constant weight. E.g . The moisture content of digitalis and ergot should not be more than 5%W/W, respectively.

31. 2) S olubility- Drug constituents have specific behaviour towards solvents are taken into consideration. Eg . Solubility of colophony of colophony in light petroleum, the solubility of balsam of Peru in solution of chloral hydrate. 3) optical rotation- An isotropic crystalline solids and samples containing an excess of one enantiomer of a chiral molecule can rotate the orientation of planepolarized light. Such substances are said to be optically active, and this property is known as optical rotation. Eg . Eucalyptus oil (0o c to +10o c), honey (+3o c to -15o c ) 4) Refractive index- It is defined as the property of a material that changes the speed of light, computed as the ratio of the speed of light in a vacuum to the speed of light through the material. when light travels at an angle between two different materials, their refractive indices determine the angle of transmission refraction of the light beam E.g. castor oil 1.4758-1.527

32. 5) Specific gravity- It is also known as relative density. The ratio of the mass of a solid or liquid to the mass of an equal volume of distilled water at 4o c(39o F) or of a gas to an equal volume of air or hydrogen under prescribed conditions of temperature and pressure. Eg . Specific gravity of drugs are cottonseed oil 0.88-0.93, coconut oil 0.925, castor oil o.95,etc. 6) Viscosity- Viscosity of a liquid is constant at a given temperature and is an index of its composition. Eg . pyroxylin kinematic viscosity, 1100-2450 centistokes. 7) Melting point- Plant constituents have very sharp and constant melting points. As far as crude drugs are concerned, melting point range has been fixed due to the mixed chemicals. Eg . Beeswax 62-65o c,wool fat 34-44o c 8) Ultraviolet light Certain drugs fluorescence when the cut surface or the powder is exposed to ultraviolet radiation, and it is useful in the identification of those drugs . Eg . Some pieces of rhapontic , Indian and Chinese rhubarb are very difficult to distinguish, and it is very difficult in powdered form, but examination in ultraviolet light gives such marked differences in fluorescence that the varieties can be easily distinguished from each other.

33. 9.) Ash value- The residue remaining after incineration is the ash content of the drug. Significance- Ash value is an important parameter to prove acceptability and purity in case of drugs that are collected or stored by incorrect way. High ash value is indicative of contamination, substitution, adulteration in crude drug . E.g . Inorganic salts, naturally occurring in drug in the form of adulteration. Ash value is determinant of identity or purity of drug a. Acid insoluble ash : It determines amount of silica present, especially as sand siliceous earth b. Water soluble ash : It determines the amount of water soluble constituents in crude drug like tannin, sugar, plant acids, mucilage, glycoside etc.

34. 10) Extractive values - The extracts obtained by exhausting crude drugs with different solvents are approximate measures of their chemical constituents. Various solvents are used according to the type of the constituents to be analysed . - “Water soluble extractive” is used for crude drugs containing water-soluble constituents like glycosides, tannins, mucilage etc ; - “Alcohol- soluble extractive” is used for crude drugs containing tannins, glycosides, resins , etc . - “Ether-soluble extractives” are used for drugs containing volatile constituents and fats. 11) Foreign organic Matters- The parts of the organ or organs other than those parts of drugs mentioned in the definition and description of the drug are known as foreign organic matters. They may be insect, moulds , earthy material, animal excreta, etc. E.g . Garlic should not contain more than 2%, saffron should not contain more than 2%.

35. Biological method e valuation When the estimation of potency of crude drug or its preparation is done by means of its effect on living organisms like bacteria, fungal growth or animal tissue or entire animal, it is known as bioassay. Some drugs have specific biological and pharmacological activity which is utilized for their evaluation. Actually this activity is due to specific type of constituents present in the plant extract. For evaluation the experiments were carried out on both intact and isolation organs of living animals. With the help of bioassays, strength of drug in its preparation can be evaluated. Biological evaluation or bioassay can be carried out on following living creature’s like- microbial strain (bacteria, fungus, protozoa or virus), aquatic animal (zebra fish), developed mammals ( swiss albino mice, wistar albino rat, guniea pigs, himalayan white rabbit, cat, dogs and monkeys), bird (Pigeon).

36. Living models use for biological e valuation Fungal strain Bacterial strain Guniea pig Swiss albino mice W istar albino mice Albino Rabbit Z ebrafish  ( Danio rerio ) Pigeon

37. Instrumental method e valuation Instrumental method of analysis is one of the important and advance part of quality control and standardization of herbal drug in analytical pharmacognosy . It consist to find identity, purity, content percentage and concentration of the product with respect of quality. This method is consist of various instrumental tools and technique to check the quality control and standardization of herbal drugs. This method is use when the all the evaluation parameter gets fail to predict the quality report the instrumental use for it to know the chemical and physical parameters and quality check Instrumental analytical technique using instrumental techniques such as thin layer chromatography, HPLC, GC–MS, LC–MS, Infrared (IR), and spectrophotometer , etc.

38. Chromatography technique Thin layer chromatography (TLC) High performance thin layer chromatography (HPTLC) High performance liquid chromatography (HPLC) Liquid chromatography – mass spectroscopy (LC-MS) Gas chromatography – mass spectroscopy (GC-MS) Spectroscopy technique Ultra voilet spectroscopy (UV-visible) Infra red Spectroscopy (IR) Mass Spectroscopy (MS) Nuclear Magnetic resonance (NMR) Instrumental method of quality control

39. SPECTROSCOPICAL ANALYSIS :- Uv -visible spectroscopy – UV/Vis spectroscopy is routinely used in analytical for the quantitative determination of different analytes , such as transition metal ions, highly conjugated organic compounds, and biological macromolecules. Spectroscopic analysis is commonly carried out in solutions but solids and gases may also be studied . M ass spectroscopy - can be used to classify unknown substances by molecular weight measurement, to measure known compounds, and to determine the structure and chemical properties of molecules. Due to its capability to distinguish between substances, Mass spectrometry is used to determine unknown substances . Infrared spectroscopy ( IR spectroscopy or vibrational spectroscopy) is the measurement of the interaction of infrared radiation with matter by absorption, emission, or reflection. It is used to study and identify chemical substances or functional groups in solid, liquid, or gaseous forms. NMR spectroscopy has many applications in modern science. As we’ve explored, its primary function is analysing molecule structure and shape. However, it is also used for the following purposes like d etermining protein folding, drug screening and design.finding out how molecules interact in chemical reactions, determining the proportion of solids and liquids in lipids .

40. Example’s of herbal drug represent Spectroscopic response Examples of drugs with their uv - visible wavelength :- Herbal drugs UV range (nm) Morphine 284 Aloe- emodin 225,258,279,287,430 Caffeine 243,326 Scopholine 227,250,288,339 Amines - 3300-3500 cm -1 Alkanes- 2940-2860 cm -1 Carboxylic acid- 3520 cm -1 Cynide - 2225 cm -1 Hydroxyl- 3400-3500 cm -1 Examples of drugs with their IR frequency :-

41. CHROMATOGRAPICAL ANALYSIS :- 1 Thin Layer Chromatography TLC is one of the most important tool for separation of compound. It is widely used technique of chromatography. It is based on principle of adsorption . In this method stationary phase is a finely divided solid and it is applied as a thin layer on supporting plate and the mobile phase is a liquid which is allowed to flow on the surface of the plate by capillary action. Common adsorbent material used- Silica gel, Alumina , Kieselguhr . Drug Adsorbent Solvent system Rauwolfia alkaloids Silica gel 60 F254 Ethyl acetate: Methanol: Water (100:13.5:10) Colchicum alkaloids Silica gel 60 F254 Ethyl acetate: Methanol: Water (100:13.5:10) Founiculum valgare Silica gel 60 Tolune:Ethyl acetate(93:7)

42. . 2 HPTLC: In High Performance Thin Layer Chromatography a layer thickness of 100-150 micron is used to achieve separation. HPTLC uses open layers of adsorbents on plates or foils to separate component of samples but the quantification is observe under the UV visible chamber to observe the spot of absorbance. Significance of HPTLC: Identification and detection of adulterants in herbal product and it is also important in identification of pesticide content, myco -toxins and in quality control of herbs and health foods Drug Example - Flavonol Glycosides . Stationary Phase: Silica gel Mobile Phase: Chloroform : Benzene: Ethanol: Acetic acid : Water (11:4:2:1:2 ) Detection : Spraying with 8% AlCl 3 in ethanol. Quantification : UV absorbance 370nm.