THE INDUSTRIAL ASPECTS OF MANUFACTURING

& STANDARDIZATION OF BOTANICALS

ROOPAK KUMAR, PH. D.VICE-PRESIDENT

M/S MULTANI PHARMACEUTICALS LIMITED (DELHI, ROORKEE AND PARTNERS)MULTANI RESEARCH FOUNDATION

ANALYTICAL DIVISION (GOVT. APPROVED & NABL ACCREDITED TESTING LABORATORYEXPERT MEMBER- CHEMISTRY (AYURVEDIC PHARMACOPOEIA COMMITTEE)

OBJECTIVE OF AYURVEDIC DRUG MANUFACTURERS

Safe, Efficacious, affordable and easily available drug products to needed ones without threatening the survival of wild fauna and flaura & complying to the regulatory requirements.

AN INTRODUCTION TO AYURVEDIC MEDICINESClassical Text Books•Charaka-Samhita

•Sushruta-Samhita

•Sarngadhara samhita

•Bhavaprakasa

•Ayurveda Sar Sangraha

•Bhaisajya Ratnavali

•& many more (58+)

Proprietary•Concept derived from Indian Classical Text Books or National/International Research based knowledge.

•Formulations (Tablet/Capsules/Syrup etc.) are being development according to the need of modern life style.

QUALITY OF BOTANICALS• Authenticated Herbs up to species level preferably cultivated source.• Specified from Specified Plant Parts only.• Free from Foreign matter (Plant parts other than recommended ones. Free from

visible Fungi & algae)• Free from excessive moisture.• Specified soluble extractive (Water and Alcohol).• Volatile oil, Tannins, Saponins, Flavonoids, Alkaloids etc.,• Total ash & Acid Insoluble..• Chemical constituents should be well within specified limits.• Free from Heavy metals (e.g., Pb, Cd, Hg and As)• Free from Microbiological contaminations (TBC, TYMC and Pathogens)• Free from Aflatoxins.• Free from Pesticides Residues or within permissible limits.

SOURCING Vendors should be educated for specific requirements. Vendor’s audit. Shade drying or Direct Sun Drying should be specified. Collection of Herbs in specified seasons only. Geographical variance should be recorded for the study of chemical constituents. Storage should be according to the nature (Physical and Chemical characteristics)

of Herb. De-dusting and removal of Foreign matters. Reduction of Size to specific application without loosing characteristic useful

constituents.

CHALLENGESNational Commitment to “The CITES”

Several Herbs under CITES

Appendices I

Most endangered , threatened with extinction, International trade is banned.

Appendices II

Not necessarily endangered now but likely to be so, if trade is not closely controlled. Therefore export permit is required.

Appendices III

On the request of Party/ies, who regulates its trade. Allowed only on presentation of the appropriate permits or certificates

Expectations according to Allopathic systems (CIS countries)

WHO GMP vs Schedule T

Composition confirmation

Validation (Analytical method and Process), Stability Studies and degradation products..

QUALITY CONTROL• Authentication of Herbs by Expert/Botanist.• Certified Reference (e.g., NISCAIR, New Delhi) Standard should be used for comparison.• Identification (Chemical reactions, TLC, HPLC, UV/VIS spectroscopy, AAS/ICP, HPLC, GC, DNA Finger Print Profile

etc.,)• Markers (Characteristic phyto-constituents or Bio-active phyto-constituents) should be monitored.• Soluble Extractives• Ash content (Total ash and Acid Insoluble ash)• Assay for Chemical groups by Titrimetric/Gravimetric/Colorimetric methods, • Assay Markers based..• Enzymatic Assay• Contaminations

a) Foreign matter; should be controlled through strict Quality Control.b) Heavy metals (e.g., Pb, Cd, Hg and As) must be controlled. High HM Contamination is a Challenge to

remove except rejection..c) Microbiological contaminations (TBC, TYMC and Pathogens); if observed should not used for Churna and

Prakshep or solid dosage forms.d) Aflatoxins containing herbs should be avoided.e) Pesticides Residues should be controlled.

MANUFACTURING TECHNIQUES CONTD….

Solid Preparationsa) Tablets and Capsules

Reduction of Particle Size through Pulverizer/Chipper. Preparation of Extracts (Aqueous or Alcoholic or Hydro-alcoholic etc.,) and drying

(Validation of Process through Standardization techniques). Controlled environmental conditions (Filtered Air, Humidity and Temperature) Addition of permitted additives as per current guidelines. Lubrication and granulations. Compression or Filling, according to requirements. Packing and storage.

b) Churna or other powders Reduction of particle size and packing.Challenges: •Microbiological contaminations. Short shelf life. Preservatives normally don’t work.

MANUFACTURING TECHNIQUES CONTD….

Liquid Preparations•Syrup

Reduction of Particle Size through Pulverizer/Chipper. Preparation of Extracts (Aqueous or Alcoholic or Hydro-alcoholic etc.,) and

drying (Validation of Process through Standardization techniques). Sugar syrup preparation. Addition of permitted preservatives and coloring matter. Filtration through specified filters (Sparkler etc.,) Filling through manual or Automatic machines under controlled environmental

condition. Packing and storage.

Advantages •Better process. No Microbiological contamination. Shelf life Better.

MANUFACTURING TECHNIQUES CONTD….

Liquid Preparations•Asava/Arishtas (A wide range of effective Ayurvedic medicine)

Reduction of Particle Size through Pulverization/chipping. Jaggery powder syrup preparation. Addition of fermentation triggering aids (Dhataki puspa) Woodfordia fruticosa

Kurz flowers Optimization of Process (6-10 days) at optimized temperature. Control of Alcohol formation. Termination of reaction. Clarification, Filtration, Packing and storage.

Advantages •Better process. Homogeneity better. Microbiological contamination well controlled. Older is better.

MANUFACTURING TECHNIQUES CONTD….

Semi-solid Preparations•Avleha (A mega product “Chyawanprash” a boon of Ayurveda or Majoon in Unani)

Reduction of Particle Size through Pulverization/Chipping. Herbal Aqueous extraction. Sugar & Honey syrup preparation. Mixing of Amla Pishti/Dry Fruit pulp, Herbal Extraction and Sugar solution. Addition of Prakshep dravya (Dry Herbal powder for aroma and other curative

effects) Filtration to remove any unwanted material. Packing and storage.

Advantages •Better process. No Microbiological contamination. Best product.

NEED OF STANDARDIZATION OF BOTANICALS• Authentication through Chemical & Instrumental methods.• Monitoring of Characteristic constituents/Marker compounds.• Investigation of adulteration.• Development of specifications to release/reject the materials.• Assigning the storage conditions and period for storage of Raw materials/Finished

Goods.• Support in the optimization of process parameters (Drying, Pulverization,

extraction etc.)• Support in Stability Studies of Final Products to assign the shelf life.• Consistent Quality of final products.• Investigation of complaints.

TECHNIQUES FOR STANDARDIZATION• Potentiometric titrator (Organic acid e.g., Total Boswellic acids, Phenolic

compounds, Alkaloids etc.)• Thin Layer chromatography/High Performance Thin Layer chromatography.• High Performance Liquid Chromatography (UV/VIS, PDA, ELSD, RI, MS)• Gas Chromatography (FID, ECD, MS)• Spectroscopy Atomic Absorption Spectroscopy. Inductively Coupled Plasma- Optical Emission/Mass Spectroscopy.• Polymerase Chain Reaction (PCR)/Real Time-PCR for DNA based investigation

CRITERION OF SELECTION OF METHODS

• Compendia/Pharmacopoeia Products No compromise. Either invest or get outsourced.

• Proprietary products Evaluation of Characteristic constituents of Botanicals/Marker compounds

• Contaminations (A mandatory requirement for all)Inorganic (Heavy metals)

Organic (Aflatoxins, Pesticide residues)

Microbiological (Bacteria, Fungi and Pathogens)

Note: Poly Aromatic Hydrocarbons (PAH) and Poly Chlorinated Biphenyl (PCBs) are also in check list of Export.

MINIMUM REQUIREMENTS FOR A LABORATORY

Equipment Tests RemarksBinocular Microscopic, Dissecting microscopes, Microtome,

Microscopic Identification

Mandatory requirements

Analytical Balance, Electric Hot air oven, pH meter, Polarimeter, Melting point apparatus, Karl Fischer apparatus, Muffle furnace, Refractometer, Tablet Disintegration apparatus, Magnetic stirrer, Water bath, Desiccators, Heating mantle, Hot plate, Dean & Stark apparatus, Volatile oil apparatus (Light & Heavy), Soxhlet extraction apparatus, Alcohol determination apparatus, Boiling point apparatus, Burette, Pipette, Volumetric flasks, TLC applicator or Capillary tube (Marked for µl), TLC chambers

Chemical tests Mandatory requirements

Laminar Air Flow, Incubator, Serological water bath, Hot Air Oven, Autoclave, Microscope (High power) , Colony counter, pH meter, Analytical Balances

Microbiological contaminations

If arrangement is not available, it can be outsourced

TLC applicator, Saturation Chamber, Syringes or capillary tubes Aflatoxins or HPLC equipped with Fluorescence detector

If arrangement is not available, it can be outsourced

Atomic Absorption spectrophotometer with Hydride vapor generation Kit, Flame and Graphite Furnace

Heavy metals (Pb, Cd, Hg and As)

If arrangement is not available, it can be outsourced

Gas chromatograph equipped with Mass spectrophotometer Pesticides Residues If arrangement is not available, it can be outsourced.

MARKERS

• Marker; a characteristic Phyto-constituent may be a bioactive compound or may not be!

• Shouldn’t be common between/among two or more Herbs in a formulation.

• Bioactive Phyto-constituents should be preferred. • Should be preferably stable compound.

STANDARDIZATIONSelection of marker compound/s. Selection/Development of (preferably) Chromatographic method/s.Optimization of Extraction medium and methodologies.Chromatographic (HPTLC, HPLC and GC) methods are most suited/preferred.Method validation (Specificity, Dilution Stability, Linearity, Range, Accuracy, Ruggedness & Robustness)Analysis of RM samples.Analysis of Finished Products.Data interpretation and further work up if required.Finger print profile in case of markers are not available.

METHOD DEVELOPMENT• Chemistry of Target compound.• Molecular weight, Solubility, Polarity, Functional group, Chromophoric or Non

Chromophoric • Selection of technique/method.• Detection/Quantitation Range and accuracy level required.• Chromatographic (GC/HPLC and detection FID/ECD/MS, UV/VIS, RI/ELSD/Mass.• Column selection.• Mobile phase.• Diluent and Extraction media.• Optimization of Extraction techniques (Solvent/Solvent, SPE etc.).• Removal of interfering compounds (Filtration, Centrifugation, Adsorption etc.)• Analyte’s response (Linear response if not Linear use of Mathematical transformation)• Spiking and recovery.

SELECTION OF TECHNIQUEType of Compounds Techniques Concentrati

onAccuracy

Organic, Volatile, Non-Chromophoric, stable at higher temperature

Gas chromatograph with Flame Ionization Detector, HPTLC

>0.5% RSD < 2%

RSD >2%Organic, Highly chlorinated compounds Gas chromatograph with Electron

Capture Detector, HPTLC

>0.5% RSD < 2%

RSD >2%Organic, Non-volatile, Chromophoric, Stable or unstable at higher temperature

HPLC with UU /VIS Detector, HPTLC

>0.5% RSD < 2%RSD >2%

Organic, Non-volatile, Non-Chromophoric, unstable or stable at higher temperature

HPLC with RI/ELSD Detector/MSHPTLC

>0.5% RSD < 2%RSD >2%

Organic, Non-volatile, Chromophoric or Non- Chromophoric, unstable or stable at higher temperature

LC/MS µg level RSD < 2%

Organic, Volatile, Non-Chromophoric or Chromophoric , stable at higher temperature

GC/MS µg level RSD ≤/≥ 2%

Inorganic elements Titration/Flame Photometer/AAS/ICP/Voltametery

µg level to % Subjective to technique

Chromophoric functional group based molecules

UV/VIS Spectroscopy µg level to % RSD < 2%

REFERENCESThe Ayurvedic Pharmacopoeia of India Part-I Volume-I Standards for Single Drugs 1990

The Ayurvedic Pharmacopoeia of India Part-I Volume-II Standards for Single Drugs 1999

The Ayurvedic Pharmacopoeia of India Part-I Volume-III Standards for Single Drugs 2001

The Ayurvedic Pharmacopoeia of India Part-I Volume-IV Standards for Single Drugs Edition-I 1st Jan 2004

The Ayurvedic Pharmacopoeia of India Part-I Volume-V Standards for Single Drugs Edition-I 1st Jan 2006

The Ayurvedic Pharmacopoeia of India Part-I Volume-VI Standards for Single Drugs Edition-I 1st Jan 2009

The Ayurvedic Pharmacopoeia of India Part-I Volume-VII Minerals and Metals Edition-I 1st Jan 2009

The Ayurvedic Pharmacopoeia of India Part-I Volume-VIII Standards for Single Drugs Edition-I 1st Dec 2011

The Ayurvedic Pharmacopoeia of India Part-II Volume-I Formulations (Avleha, Curna, Ghrta, Guggulu, Gutika, Ksara/Lavana, Taila, Lepa)

Edition-I 1st Jan 2008)

The Ayurvedic Pharmacopoeia of India Part-II Volume-II Formulations(Asava & Arista, Avaleha, Curna, Ghrita, Guggulu, Taila)

Edition-I 1st Jan 2009

The Ayurvedic Pharmacopoeia of India Part-II Volume-III Formulations(Arka, Avaleha, Curna, Ghrta, Guggulu, Taila, Vati)

Edition-I 1st Jan 2011

Thin Layer Chromatographic Atlas of Ayurvedic Pharmacopoeial Drugs

Part-I Volume-I Edition-I

Macroscopic and Microscopic Atlas of Ayurvedic Pharmacopoeial Drugs(The Ayurvedic Pharmacopoeia of India)

Part-I Volume-V 2009

Protocol for Testing of Ayurvedic, Siddha & Unani Medicines

March 2007

Quality control Manual for Ayurvedic, Siddha & Unani Medicine

June 2008

REFERENCES (OTHERS)Indian Pharmacopeia Single Herbs & Extracts.

British Pharmacopoeia Monographs on Herbs, Extract and Formulations.

United State Pharmacopoeia Monographs on Herbs, Extract and Formulations.

American Herbal Pharmacopoeia More than 31 monographs. Specimen

Indian Herbal Pharmacopeia Monographs; was best presentation.

Standardization of Botanicals Good guidance.

Plant Dug Analysis TLC/HPTLC techniques.

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POTENTIOMETRIC TITRATOR

POTENTIOMETRIC TITRATOR

VISCOMETER-BROOKFIELD

HPLC WITH UV/VIS DETECTOR

ATOMIC ABSORPTION SPECTROPHOTOMETER EQUIPPED WITH FLAME, GRAPHITE FURNACE AND HYDRIDE VAPOR GENERATION ASSEMBLY

GAS CHROMATOGRAPH WITH HEAD SPACE ASSEMBLY.

Multani Pharmaceuticals Limited(Analytical Division)

Thank youQ?