Quality control & Standardization of herbal drugs

1. Quality control & Standardization of herbal drugs P. Pushpangadan National Botanical Research Institute (Council of Scientific &Industrial Research), Rana Pratap Marg, Lucknow-226001 (19th July 2003)

3. 21st Century 21st century is the century of Biology powered and propelled by scientific knowledge and technological expertise • Three technologies namely • Biotechnology • Herbal technology • Information technology (Bioinformatics) • are going to be the most powerful elements that are crucial for prosperity and welfare for the people of nations.

4. Herbal technology All technologies for the manufacture of value added plant products can be called as herbal technology • Herbal drugs and pharmaceuticals, • Nutraceuticals, • Functional foods, designer foods or health foods and health drinks • Cosmaceuticals • Biocontrol agents • Biopesticides

5. Some features of international health care • Health care policies largely market driven by the pharmaceutical industry diverting attention from health preservation to illness cure • Prevention and eradication of diseases undermines the economic basis of this industry • No satisfactory drugs available for most of the degenerative disorders characteristic of graying population and for re-emerging resistant infections • Many currently used modern drugs do not have valid proven clinical utility. • USA has among the highest per capita annual expenditure on health care ($3600) but still about 15% population is denied even basic care facilities (the best Indian state like Kerala have an annual per capita expenditure ~ $15)

6. Herbal drugs in international health care • Economic aspects: Global market of herbal drugs, Nutraceuticals ~$60 billion with 6% annual growth rate. Major share of Chinese and Koreans. Indian share variously estimated at 0.35-3.0%. Chinese production increased 200% between 1995-1999. • Local acceptance: • Developed • USA: 42% use CAM spending over 29 billion US$ and 629 million visits in 1998. • UK: 28% use, spent 1.6 billion pounds and 127 million visits in 1998. • Australia: 60% use, A$ 620 billion in 1999. • Developing • Malaysia: Per capita consumption of traditional drugs, more than double of modern pharmaceuticals. • S.Korea: Per capita consumption of traditional drugs 36% more than modern drugs. • African countries: 9 to 10 patients attending hospital OPD have first consulted a traditional healer.

7. Advantages of herbal drugs • Modern drugs can produce serious side effects • Latrogenic diseases fourth leading cause of death in USA and other developed nations (JAMA, April 1998). • Side effects of drugs kill more Americans annually than the world war II and Vietnam war combined (M. Rath N. Y. Times 28.2.2003). • Around 2600 persons died in the Twin Tower tragedy on 11th September 2001 causing global repercussions. It is, however, not recognized that about the same number die in USA from side effects of prescription drugs every 10 days (JAMA, April 1998).

8. Herbal drugs are best suited for: • PRIMARY HEALTH CARE • INFECTIOUS DISEASES • AIDS and other viral infections • Opportunistic infections • MDR infections (e.g. T.B., Malaria) • DEGENERATIVE & GERONTOLOGICAL CONDITIONS • Osteoporosis • Chronic arthritis like osteoarthritis and rheumatoid arthritis • Neurological like Alzheimer, Parkinsonism • Anti-aging • Metabolic disorders • Diabetes • Dyslipidemias • Other conditions • Microcirculatory disorders • Liver diseases • Immunostimulants • Anti-cancer • Drugs affecting male libido

9. Herbal Drugs • Inclusions in Pharmacopoeia: • Chinese Pharmacopoeia: 1997 edition has 647 traditional drugs. • European Pharmacopoeia: 2000 edition contains monographs on 152 crude drugs. • Indian Pharmacopoeia: 1996 edition number shrinked to 57 including only 12 crude drugs. • Inclusion in essential drug list: • None in India. • In Shanghai hospitals: • 500 herbal drugs in essential drugs list of about 1000. • Expenditure on drugs decreased from 67% of hospital budget in 1992 to 51% in 1996. • Growth rate of drug expenditure decreased from 23.4% to 0.3% for outdoor and from 28.2% to 2.4% for indoor patients. • Approximate annual saving of 600 million US$ in 7 hospitals. (Hu, 2001)

10. The global flora, an unfathomed ocean • GLOBAL FLORA: • About 2 million flora & fauna scientifically named. Estimates up to -10 million • Over 50% plants. -45000 species of angiosperms only • Only 5-15% of plants properly studied for biological activity (Cragg et al, 1997) • One in 125 plant species contains useful pharmaceutical (Callahan, 1996) • MEDICINAL PLANTS: • WHO compiled an inventory of 21,000 plants used for medicinal purposes in 91 countries (Penso, 1983) • Less than 10,000 species have been investigated (Bhatt, 1997) • THE INDIAN SCENE: • Over 17 ,500 species, many endemic • Traditional systems of medicine use 2,000; Ayurvedic medicines alone need 800 species • Folklore use of 8,000 plants reported. Includes most plants used in traditional systems also CSIR has screened about 4000 species but few other broad based studies

11. Traditional heritage India is a mega-diversity country rich in all three levels of biodiversity species, genetic and ecosystem/ habitat. India is also rich in cultural diversity with a history of over 6000 years. India’s medical heritage is most important heritage. • Organized, codified and systematically arranged written traditions with conceptual philosophy and rationales like Ayurveda, Siddha, Unani and Amchi use almost 2000 plant species • Oral traditions – practiced by village physicians, folk healers, tribal healers – called as local health tradition use over 8000 plant species

12. India’s strength in Herbal Technology 8000 Medicinal Total 10000 species 325 3500 Edible 425 Pesticides 1000 Others 550 Fibre Gums, Resins & Dyes

13. 900 sp. Ayurveda 700 sp. INDIAN SYSTEMS OF MEDICINE Unani 600 sp. Siddha 250 sp. Amchi 30 sp. Modern 8000 species India’s strength in Herbal Technology THE INDIAN FLORA (ca 17500 species)

14. Bioprospecting • Gene Prospecting • Genetic engineering • Crop development • Fermentation • Cell culture • Chemical Prospecting • Drugs and pharmaceuticals • Pesticides • Cosmetics • Food additives • Other industrially valuable • Chemical products • Bionic Prospecting • Designs • Sensor technologies • Architecture • Bioengineering • Biomodeling

15. Bioprospecting: Linkages and leads Biodiversity & IK/TK Biotechnology Bioprospecting Information technology • Drug development • Pharmaceuticals • Agrochemistry • Cosmetics • Proteins • Enzymes • New crop varieties • GMOs • GM foods • Designs etc. Herbal technology Conservation Sustainable use Benefit sharing Bioinformatics IPR

16. Standardization of Herbal drugsRaw Drugs • Passport data of Raw Plant Drugs (Crude drugs) • Correct taxonomic identification & authentication • Study on the medicinal part: root, stem, bark, leaves, flowers, fruits,nuts, gum, resins etc. • Collection details: Location, stage & development/ growth of the plants, time, pre-processing storage etc. • Organoleptic examination of raw drug: • Evaluation by means of sensory organs: touch, odour taste • Microscopic & molecular examination • Chemical composition (TLC, GLC, HPLC, DNA fingerprinting) • Biological activity of the whole plant • Shelf life of raw drugs

17. Standardization of Herbal drugs-Herbal Formulation • Follow defined Good Manufacturing Practices (GMP) • Scientific Verification • Toxicity evaluation • Chemical profiling • Pharmacodynamics – effect of drug in the body • Pharmacokinetics – absorption, distribution, metabolism, mechanism of action and execution • Dosage • Stability and shelf life • Presentation and Packing • Therapeutic merits – Compared with other drugs

18. Good Practices/Techniques in Herbal Products • Good Survey of literature (Ancient & Modern) • Develop and Observe Norms of: • Good Agricultural Practices (GAP) • Good Collection/Harvesting and Post Harvest Handling Practices (GCP/ GHP & GPHP) • Good Laboratory Practices (GLP) • Good Clinical Practices (GCP) • Good Manufacturing Practices (GMP) • Good Marketing Techniques (GMT)

19. Physico-chemical value Parameters required for quality evaluation of herbal drugs

20. Standardization & Quality Evaluation of Herbal drugs • Shape • External • Marking • Qualitative • Quantitative • SEM Studies • Powder Studies • Colour • Odour • Taste • Texture • Fracture Macroscopic Microscopic BOTANICAL ORGANOLEPTIC • Moist. Cont. • Extrac. Values • Ash Values • Fluores. Analy. QUALITY EVALUATION OF HERBAL DRUGS PHYSICAL Microbial Contamination BIOLOGICAL CHEMICAL • Toxicological • Pharmacological • Other specific activities HPTLC Finger printing Sec. Metabolites DNA Finger printing • Qualitative • Quantitative • Chromatography • Heavy metal • Pesticide residue • Mycotoxin Antagonistic HPTLC GLC HPLC • Bacterial • Fungal

21. THE AYURVEDIC THERAPEUTIC STRATERGY • Determine PRAKRUTI (Constitution) by -history taking -observations • NIDANA (Diagnosis) Nature, degree and extent of imbalance of Tridoshas. Library of 5800 clinical signs and symptoms in Ayurvedic texts • CHRONOBIOLOGY: Impact of season, time and environment on Tridoshas. • SWASTHAVRUTA: Life style modification • AHARA: Dietary modifications • PANCHAKARMA: Purification of the body • AUSHADHI: "Designer Medicine" unique for the particular patient prepared from a Pharmacopoeia utilising 1200 plants, 100 minerals and 100 animal products in numerous formulations.

22. OBJECTIVES OF RESEARCH ON AYURVEDIC DRUGS • Improved formulations and reduced number of Ayurvedic drugs • Use of GMP procedures and QC • Certified shelf life and improved dosage form • Validated indications and contraindications • Deletion of obsolete or toxic formulations • Use of Ayurvedic drugs in modern clinical practice • Inclusion in essential list of drugs • Adjunct to existing drugs • Treatment of diseases where modern drugs not available or unsatisfactory • Development of suitable formulations, standardized extracts or active constituents • IPR protection wherever feasible • Inclusion in Pharmacopoeias.

23. OBJECTIVES OF RESEARCH ON AYURVEDIC DRUGS • New indications for Ayurvedic drugs • Development of new drugs for Ayurvedic practice • Utilizing leads from other countries • Study of unscreened flora, specially endemic or threatened species • Studies on Ayurvedic drugs for veterinary use

24. Development of new drug/ novel uses for Ayurvedic drugs • Utilization of new leads on natural products from other countries. Several of these plants or related species are found in India • The vast unscreened flora of the country and published activity data on some of these • Study of further accessions from families yielding active plants

25. Raw drug standardisation (Example) Maramanjal (Root and Stem) - Coscinium fenestratum(Gaertn) Colebr. Maramanjal consists of the dried root and stem of Coscinium fenestratum(Gaertn) Colebr. (Fam. Menispermaceae); a large woody climber with stout stem and branches, occuring in the hills of Malabar region, particularly Western Ghats. a) Macroscopic : Root -5 to 30 cm or more in length, 2 to 5 cm. in diameter, somewhat longitudinally grooved, transversely cut surface smooth, yellow; texture rough and fibrous; acrid in taste; no particular odour. Stem -15 to 30 cm. or more in length, 2 to 8 cm. in diameter, straight or occasionally slightly twisted, pale grey or greyish yellow with a fairly smooth surface, marked with longitudinal striations spaced about a mm apart, cut surface yellowish-green to yellow in colour showing wedge shaped areas, fissured with shallow vertical slits of varying length; texture hard, acrid in taste.

26. b) Microscopic : Root -Transection almost circular in outline; cork cream coloured, 20 to 30 or more rows of uniform rectangular cells with 1 to 2 stone cells; outer cortical tissue characterized by the presence of very prominent yellowish band almost in the form of ring of thick walled, pitted stone cells; prismatic crystals of calcium oxalate found in the thick walled cells; phloem in groups, sieve tubes with oblique pore and simple perforation plate; narrow radiating wedge shaped xylem strips; medullary rays multiseriate with thick walled cells and wedge shaped, extending to the centre, cells filled with rod shaped crystals of calcium oxalate and starch grains which are circular, appearing lenticular on edge view, simple, 30-45 m in diameter hilum indistinct or dot-like centrally placed if present, lamellae indistinct; vessels filled with tyloses and in mature root these tyloses become thick walled giving the appearance of stone cells; fibres long, lignified. Stem - The transverse section circular in outline, shallowly crenate; cork 20 to 40 cells thick; cortex 5 to 8 layers of tangentially elongated parenchymatous cells having very conspicuous yellowish crenate bands of hard tissue or stone cells with radiating canals and filled with dark yellow content and almost capping the wedge shaped medullary rays and phloem; sclerotic elements cubical to oval with very thick pitted walls filled with prismatic crystals of calcium oxalate; phloem distinct; xylem narrow, radiating, wedge shaped as in root, vessels 70 to160 m in diameter, solitary, transversely elongated, pitting reticulated with small lenticular orifices, occluded with thick walled tyloses; fibres septate to nonseptate, septate fibres having 2 to5 septa, 270 to 400 m long and 12 m in diameter; medullary rays extend from pith to periphery, broad, multiseriate, interfasicular,15 to many cells high and 2 to many cells wide; pith consist of two regins (i) parenchymatous cells circular to polyhedral in shape with intercelular spaces, cells larger towards the centre (ii) 4 to 6 layers of smaller collenchymatous cells towards periphery.

27. Starch grain Powder -Powder of both root and stem yellow with greenish tinge, bitter and odourless. Microscopical examination shows the presence of fibres, tyloses, stone cells, prismatic crystals of calcium oxalate, starch grains circular appearing lenticular shaped on edge view, simple, 30-45 m in diameter hilum indistinct or dot like centrally placed if present, lamellae indistinct and fragments of vessels, tracheids and parenchymatous cells; when treated with 1N NaOH aqueous and mounted in nitrocellulose in amylacetate emits very characteristic canary yellow colour under UV-365 nm.

28. IDENTITY, PURITY AND STRENGTH – Root– Foreign matter : Not more than 1% Moisture content : Not more than 7% Total ash :Not more than 2% Acid insoluble ash : Not more than 0.4% Alcohol soluble extractive:Not less than 11% Water soluble extractive : Not less than 10 % Total alkaloid as berberine chloride :Not less than 2% Stem – Foreign matter : Not more than 1% Moisture content : Not more than 6% Total ash : Not more than 3% Acid insoluble ash : Not more than 2% Alcohol soluble extractive : Not less than 3% Water solubleextractive :Not less than 8% Total alkaloid as berberine chloride :Not less then 1% CONSTITUENTS – Alkaloids - Berberine, Palmitine, Jatrorrhizine, Proto-berberine, Barbarrubin, N, N-di-lindacarpine, Thalifendine and Columbamine.

29. T.L.C. – T.L.C. of alcoholic extract (methanolic) of drug on Silica Gel G plates (precoated aluminium plate) using Isopropanol : Formic acid : Water (45 : 0.1 : 0.4) shows nine fluorescent zones under UV 254 at Rf. 0.10 (blue), 0.17 (blue), 0.24 (greenish yellow), 0.34 (greenish yellow) 0.39 (green), 0.5 (blue), 0.56 (bluish green), 0.78 (blue) and nine fluorescent spots of blue colour under UV 366 nm at similar Rfs. On spraying with modified Dragendroff's reagent 4 spots appear at Rf. 0.10 (brown), 0.24 (brown), 0.34 (reddish brown-Berberin), 0.83 (brown) and 0.89 (brown). T.L.C. Finger-print profile of Methanolic extract of ‘Maramajal’ (Coscinium fenestratum) Dectetion : (a) Under UV 366nm (b) Under UV 254 nm. (c) Under visible light after spraying with detecting reagent.

30. PROPERTIES AND ACTION – Rasa : Kasaya, Tikta Guna : Laghu, Ruksa Virya : Usna Vipaka : Katu Karma : Shothara, Vednasthapana, Vernsodhan, Chakshoya IMPORTANT FORMULATIONS –Rasaut or ras, Syrups Coscinii fenestrati, Radicis Liquidum. THERAPEUTIC USES – Tikta-Usna, Dipan, Pachan, Anuloman, Pitsarak and Krimighna, Raktavardhak, Rakta shodak, Javarghan and Katupostic. DOSE -1 – 3 g in powdered from; Infusion 4-12 drams; Tincture ½ - 1 drachm; Decoction ½-1ounce. MARKET SAMPLES – Mumbai sample– I - Identified as stem of C. fenestratum Mumbai sample – II - Identified as stem of Berberis aristata Coimbatore sample - Identified as stem of C. fenestratum Delhi sample - Identified as stem of B. aristata Haridwar sample - Identified as stem of C. fenestratum Kanpur sample - Identified as Rubacious stem Lucknow sample - Identified as stem of B. aristata Note -The only two samples of Maramanjal were studied because this plant is restricted in Malabar region of the country. The second sample was procured from Sri Lanka, as this is common in Sri Lanka.

31. Important Aspects involved in Utilization of Medicinal Plants • 1.Survey, inventory, documentation ( digitization) of the Medicinal plant wealth and its associated knowledge system • Database • 2. Conservation and sustainable utilization • Conservation • Insitu • Exsitu • Seed Bank, gene/DNA bank • Tissue repository • Cryo bank • Field Gene Bank • 3.Bioprospecting • Gene prospecting • Drug prospecting • Herbal drugs • Biomolecules

32. 4. Economic Evaluation of Medicinal Plants • 5. Domestication and cultivation of Medicinal Plants • 6. Development of protocols for Agrotechniques and post harvest managements • 7. Standardization of Raw Drugs and finished products • 8. Product Development • GPC, GCP, GMP and GQC • 9. Trade • Domestic • International • 10. Developing Industrial government producer • Partnership

33. Thank You