Standardizatio n of herbal drugs

Pandure Dinesh R. Department of Pharmacology and Toxicology Mumbai Veterinary College

Pandure Dinesh R. M.V.Sc.(Sr.) Department of Pharmacology & Toxicology Mumbai Veterinary College,Parel

Introduction What is standardization? WHO Guidelines Different Tests Regulatory Bodies Conclusion References

Ayurveda
Originated in India Ayurveda
Ayus (life) Veda (knowledge or science)

AYURVEDA
PAST  Raw material collected by vaidya  Formulated the drug  Dispensed the drug and supervised the use of drugs PRESENT  Raw material supplied by suppliers  Number of manufacturers  Dispensed by hundreds of practioners

Importance of Herbals
 Limitations of Allopathy : 1.Degenerative disorders 2.Re-emerging resistant infections
 Herbal drug avenues:  Primary health care  Diabetes  Dyslipidemias  Liver diseases  Anti-aging  Osteoporosis
Immunostimulants Anti-cancer Drugs affecting male libido Chronic arthritis Neurological like alzheimer,

Pushpangadan(2003)

World Health Organization

 Identified medicinal plants: 21,000 in 91 countries
(Penso, 1983)

 Investigated: <10,000
(Bhatt, 1997)

 Economic aspects:
Global market:$60 billion (6% annual growth rate.) Chinese and Koreans Indian share: 0.35-3.0%.
Pushpangadan(2003)

Indias strength in Herbal Technology

unities (ora mm l) Co al
900 sp. 700 sp. 600 sp. 250 sp. 30 sp.

INDIAN SYSTEMS OF MEDICINE

e used b ts ar yT lan r ib lP

Ayurveda Unani Siddha Amchi Modern

8000 species

Me d ici n

What is standardization..?
 Definition of the content of a constituent or a group of substances with known therapeutic activity OR  Ensuring that every packet of medicine has correct ingredients in the correct amount and will induce the intended therapeutic effect.

Why we follow this?

 Quality control  Safety for Human consumption  World market  WHOs, (GMP) ISO 9000  Environmental safety- ISO 14000
(Ecological damage) Joy et al.(1998)

LEGAL REQUIREMENTS

USA : Proof of quality, safety and efficacy Europe: Harmonization by use of the European
Scientific Cooperative on Phytotherapy (ESCOP), since 1989

India : GMP norms should be

followed

    

Botanical: Physicochemical TLC:Pharmacological: Toxicological: Microbial contamination: -

WHO GUIDELINES WHO Guidelines

Panchawat et al.(2009)

QUALITY EVALUATION OF HERBAL DRUGS

Authentication (Schedule T of the Drugs and Cosmetics Act ,1940)

 Institute for Authentication
Central Council For Research In Ayurvedic Medicine Central Council For Research In Unani Medicine(CCRUM) Pharmacopeic Data

 Parameters
Parts of plants - leaf, flower, root, stolen Regional status Family Biological source Chemical constituents
Gautam et al.(2010)

BOTANICAL EVALUATION
Example :Nux vomica

Macroscopy parameter Colour- Greenish -brown Odour-none Taste-intensely bitter

Microscopy parameter Epidermis-strongly thickened, pitted, lignified Collapsed cells- present Endosperm- unlignified, plasmodesma, aleurone graines Calcium oxalate crystalspresent

Size-10-30mm in diameter 4-6mm in thickness

Parts of the Plant: Qualities

Alkaloid rich, often toxic, often medicinal Leaves: Nutrient rich Flowers: Some nutrients, energetically powerful Seeds: Nutrient dense, often toxic, toxins often medicinal Roots:

Under ground parts Freed from soil

Insect attacked plant parts rejected

Environmental variation
 Energy moves with the season  Roots in spring and fall when plant is storing constituents Ex...  Datura (Alkaloid): lower in cloudy/rainy weather  Volatile oils: warmer weather

Annual

Biennials Roots Flowers

Perennials Seeds

Leaves Flower
Seeds

Physical evaluation
 VISCOSITY Viscosity of a liquid is constant at a given temperature & is an index of its composition.

 MELTING POINT  Pure phytochemical: Constant MP  Their purity can be checked E.g. Colophony- 75-80c Cocoa butter- 30-33c

Contd.  SOLUBILTY : Adulteration E.g. Pure Asafoetida -soluble in carbon disulphide  MOISTURE CONTENT Decomposition (chemical change or microbial contamination) Moisture:105c in an oven E.g. Aloe 10% (w/w)  VOLATILE MATTER Toulene distillation aromatic drugs

Drugs Lemon Lemon clove

Volatile oil content (%w/w) Not less than 2.5 Not less than 2.5 Not less than 15

Contd  OPTICAL ROTATION Property of rotating the plane of polarized light(optical rotation) Normally, the optical rotation is determined at 25c using sodium lamp as the source of light. E.g. castor oil - 3.5to +6 REFRACTIVE INDEX When a ray of light passes from one medium to another of different density, then the ratio of velocity of light in vaccum to its velocity in substance is termed as refractive index of second Drugs Refractive index medium. Arachis oil 1.4678-1.470 E.g. Caraway oil 1.4838-1-4858
Castor oil Clove oil 1.4758-1.527 1.527-1.535

Contd  ASH VALUES AND EXTRACTIVES The residue remaining after incineration is the ash content of drug
 Total ash method  Acid insoluble ash  Water soluble ash

 BITTERNESS VALUE
Appetizing agents quinine hydrochloride

 WATER AND VOLATILE MATTER: Azeotropic method is used to directly measure the water present in a material Loss on drying In order to measure volatile matter, plant is diluted with water and distillate is collected in a graduated tube. The aqueous portion separates and returns to distillation flask. A solvent of low mass density with a suitable boiling point may be added to measuring tube to easily separate the volatile oil.

Chemical evaluation Qualitative chemical evaluation

Tests
Alkaloids Mayers tests Wagners tests Hagers tests Amino acids Millons tests Ninhydrine tests Carbohydrates Molischs tests Barfoeds tests Selivanoffs tests

Reagents used
Potassium mercuric iodide solution Iodine potassium solution Saturated solution of picric acid Millon reagents Ninhydrin solution Alcoholic a-naphthol+sulphuric acid Barfoed reagents Selivanoffs reagents

Color formed
Cream precipitate Brown precipitate Yellow colour White precipitate Violet colour

Purple to violet colour Rings Red colour Rose colour(keton)

Quantitative chemical evaluation

 Chemotaxonomy: Classification of plants and organisms by their chemical constituents  Techniques: Phytochemical Screening  Thin layer chromatography (TLC)  High performance liquid chromatography (HPLC)  Gas chromatography (GC)  Ultraviolet/Visible spectroscopy (UV/VIS)  Mass spectrometry (MS)  Combination of GC and MS (GC/MS)  Over-pressured layer chromatography (OPLC)

DNA fingerprinting
Identification of medicinal plants Identification of germplasms of important or endangered plants for future cultivation or conservation (Vasudevan 2004)

Biological evaluation
Bacterial and mold contents in a plant are influenced by the environment Harvesting and production practices
Permissible Limit As per WHO

Microbes

For further processing 104 Cfu/g -

for Topical use

For intravenous

E. coli Enterobacteria Salmonella Total aerobic bacteria

102 Cfu/g 104 Cfu/g 107 Cfu/g

101 Cfu/g 10+ Cfu/g 105 Cfu/g

REGULATORY BODIES (AN ORGANISATIONAL SET UP)

CENTRAL LEVEL
Central Department under Ministry of Health & Family Welfare. Govt of India Central Research Councils. Central Regulatory Bodies. National Institutes. National Medicinal Plant Board. Central Pharmacopoeia Laboratories Manufacturing Unit Hospitals & dispensaries

STATE LEVEL
AYUSH Department or Ministry. State Directorates State Boards/Councils for registration of practitioners. State Licensing/Drug Control Authorities. State colleges/institutions. State Pharmacies State Drug Testing Laboratories. State Medicinal Plant Boards

TOXICOLOGICAL STANDARDIZATION
 Determination of Pesticides. Heavy metals Radioactive contamination Aflatoxins.

Determination of pesticides.
Qualitative

: Quantitative :
1. 2. 3. 4. 5. 6. 7. 8. 9.

Partition and/or adsorption, GC, MS, or GC/MS.

Limit as per FDA/ EP

S.No Name of . Pesticides/insecticides

Quinolphos DDE Alderin Dieldrin DDT DDD HCH (Hexa chlorocyclohexane) Malathion Parathion

0.01 1.00 0.05 0.05 1.00 1.00 0.30

ppm ppm ppm ppm ppm ppm ppm

0.10 ppm 0.30 ppm MRL level ,WHO

Trace amounts dangerous Amount is estimated by matching the depth of colour with of standard stain

Determination of arsenic and heavy metals

MRL level ,WHO Arsenic Cadmium Mercury Lead 10.0 ppm 0.30 ppm 1.00 ppm 10.0 ppm

Radioactive Contamination
 Source:
Radionuclides Nuclear accident (Eg..Nuclear accident in Chernobyl)

 Effect depends on:

Level of contamination Quantity of the contaminant consumed Quantity of herbal medicine normally consumed by an individual
Shrikumer et. al.(2006)

Fungus(Aflatoxins)
Dangerous in minute amounts Faulty storage Eg.-Nuts,Cereals TLC : for confirmation(After clean-up ) Toxin produce cancer in human beings Stored nuts and cereals are contaminated by the fungus They should therefore be determined after using a suitable clean up procedure.

PHARMACOPOEIA FORMULARIES Ayurvedic Pharmacopoeia: 418 monographs Ayurvedic Formulary: 636 formulations. Pharmacopoeial laboratories of Indian medicines(PLIM) for SOPs and pharmacopoeial standards.

Conclusion
There have been various guidelines issued on the standards of the herbal drugs by the concerned governing bodies like CDSCO, US-FDA etc. While developing an herbal drug formulation, producer must have all the related knowledge. There are still many drugs which are not included in these official guidelines and books. Stringent regulation have been made to determine the conformity of these new chemicals

References
Archana Gautam, Shiv Jee Kashyap, Pramod Kumar Sharma, Vipin and Kumar Garg(2010) Identification, evaluation and standardization of herbal drugs,Der Pharmacia Lettre, 2(6): 302315 P. P. Joy, j. Thomas samuel, mathew baby, p. Skaria (1998) Medicinal plants Kerala Agricultural University WHO (2002) Traditional medicine strategy World health organization, Geneva. S Shrikumer; U Maheshwari; A Sughanti; TK Ravi(2006) WHO guidelines for herbal drugs standardization. Sunita Panchawat, Kamal Singh Rathore,.Dr.S.S.Sisodia, Dr. R.K.Nema(2009) Standardisation And Evaluation Of Herbal Drug Formulations,Articlebase

 P mukharji. Quality control methods of herbal drugs, 1st ed., Business horizon pharmaceutical, Kolkata, 2001; 578-80. CK Kokate; AP Purohit; SB(2006) Gokhale. Pharmacogognosy , 35th ed., Nirali Prakashan, Pune, 98-114. M Ali (2009)Pharmacognosy and phytochemistry 3rd ed., CBS publishers & distributors, Delhi, 181-182. PM Patel; NM Patel; RK Goyal. The Pharma Review, 2006, (22), 113. PS Sagar Bhanu; R Zafar; R Panwar. The Indian Pharmacist, 2005, 4(35), 2005, 19-22. Yoganarashimhan SN(1996) Medicinal Plants of India, Karnal for interline Publishing Pvt Ltd,Bangalore. 4. PM Patel; NM Patel; RK Goyal(2006) The Indian Pharmacist, 5(45), 26-30.

 Vasudevan H. 2004. DNA fingerprinting in the standardization of herbs and nutraceuticals. The Science Creative Quarterly. (http://www.scq.ubc.ca/?p=286) P. Pushpangadan (2003)Quality control & Standardization of herbal drugs, National Botanical Research Institute,Lukhnow

Thank you