The Industrial aspects of manufacturing and standardization of botanicals
1. 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 LABORATORY
EXPERT MEMBER- CHEMISTRY (AYURVEDIC PHARMACOPOEIA COMMITTEE)
2. 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.
3. AN INTRODUCTION TO AYURVEDIC MEDICINES
Classical 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.
4. 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.
5. 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.
6. CHALLENGES
National 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..
7. 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.
8. MANUFACTURING TECHNIQUES CONTD….
Solid Preparations
a) 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.
9. 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.
10. 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.
11. 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.
12. 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.
13. 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
14. 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.
15. MINIMUM REQUIREMENTS FOR A
LABORATORYEquipment Tests Remarks
Binocular 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.
16. 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.
17. STANDARDIZATION
Selection 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.
18. 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.
19. SELECTION OF TECHNIQUE
Type of Compounds Techniques Concentratio
n
Accuracy
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/MS
HPTLC
>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%
20. REFERENCES
The 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
21. 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.
