m.pharm analysis, pharmaceutical analysis, food analysis, cosmetic analysis and evaluation, modern bio analytical techniques, advanced pharmaceutical analysis, phamaceutical validation

1. SUBJECT: PHARMACEUTICAL VALIDATION
KARNATAKA COLLEGE OF PHARMACY
TOPIC : QUALIFICATION OF HPTLC
SUBMITTED BY: SUBMITTED TO:
ABHISHEK DR. C. SREEDHAR
M. PHARM 1ST SEMESTER PROF. AND HOD
DEPT. OF PHARMACEUTICAL ANALYSIS DEPT. OF PHARMACEUTICAL ANALYSIS
KARNATAKA COLLEGE OF PHARMACY KARNATAKA COLLEGE OF PHARMCY
BANGALURU BANGALURU
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2. CONTENTS
• INTRODUCTION
• Principle of HPTLC
• Qualification of HPTLC
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3. INTRODUCTION
 HPTLC is an advanced form of thin-layer chromatography (TLC) used for the
separation and analysis of complex mixtures of organic and inorganic compounds.
 HPTLC offers several advantages over traditional TLC, including higher resolution,
better reproducibility, and faster analysis times.
 HPTLC has a wide range of applications in various industries, including
pharmaceuticals, food and beverages, cosmetics, and environmental monitoring
 Now a day, HPTLC is more useful than TLC and HPLC. Because HPTLC is independent
of sample application, chromatogram development, detection, etc..
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4. Principle of
HPTLC
The HPTLC works on the same principles
as TLC such as the principle of
separation is adsorption. The mobile
phase or solvent flows through the
capillary action. The analytes move
according to their affinities towards the
stationary phase (adsorbent).
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5. SCHEMATIC PROCEDURE FOR HPTLC METHOD
DEVELOPMENT
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6. Qualification of HPTLC
• Qualification is a part of validation. It is action of proving and
documenting the equipment which are properly installed.
• Steps involved in qualification are:
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7. A. Design Qualification (DQ)
• This qualification verifies that the rigorous specifications and design
review methods defined in the Quality Management System of the
manufacturer have been followed.
• ISO 9001 Certified Quality Management System ascertains planned
testing procedures, error reporting and controlled updating of
documents.
• Compliance is documented e.g.by the "Declaration of System
Validation" and “ Declaration of Conformity" supplied with specific
products.
• The Design Qualification is sometimes used in a different meaning.
One common misunderstanding is to use DQ for "suitability of the
laboratory equipment".
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8. B. Installation qualification (IQ)
• The qualification is performed at the site and time of installation
• It documents that all key aspects of the installation comply with the
manufacturer's specifications, codes, safety and design parameters.
• In order to qualify for an IQ Certificate, this procedure is to be
performed by a Product Specialist by CAMAG.
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9. C. Operation Qualification (OQ)
• This qualification is performed after installation and is repeated at
certain intervals recommended by the manufacturer or defined by
the customer.
• It documents that all modules of the equipment perform consistently
throughout the specified operating ranges.
• The initial OQ is performed by the person responsible for the IQ at
installation.
• In order to qualify for an OQ Certificate, this procedure is to be
performed by a Product Specialist, approved by manufacturer.
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10. • Repetitive OQ's can be performed by a system user well aquatinted
with the system, following guidelines issued by manufacturer.
• On request of the customer, such OQ's can be performed by a
Product Specialist or Service Engineer, against a fee or within a
service contract.
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11. D. Performance Qualification (PQ)
• PQ is performed to ascertain that the instrument (system) is suitable to
perform a specific analytical task as part of the manufacturing process.
• PQ is an on-going task with the customers samples and procedures
including preventive maintenance and regular tests, such as system
suitability and quality control analyses with creation of QC-charts.
• For computer systems it also includes regular data backup, virus checks
and change control procedures.
• PQ can thus only be performed by the user himself who also has to .
create the SOP's based on the analytical task, procedure, the different
instrument manuals and the customer's QC requirements
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12. Check the performance of HPTLC in terms of
• Linearity of spotting
• Reproducibility of Spotting
• Detection capacity
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13. A. Linearity of spotting
• Apply 2ul, 4ul, 6ul, 8ul and 10 µl of solution on HPTLC plate with
spotter.
• Allow the plate to run in mobile phase.
• Dry the plate with drier.
• Scan the plate with scanner.
• Check the linearity and correlation coefficient between the spots.
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14. B. Reproducibility of Spotting
• Apply 10µl solution on HPTLC plate for six times in sequence.
• Allow the plate to run in mobile phase.
• Dry the plate with drier.
• Scan the plate with scanner.
• Calculate the Relative Standard Deviation(RSD) for six tracks.
Acceptance limit
• Documentation of RSD,
• RSD Limit: NMT 3.0%
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15. C. Detection capacity
REQUIREMENTS
I. Alumina glass plate.
II. Sodium salicylate.
III. 96% v/v alcohol
• Preparation of stock solution
Stock solution-1:-
Weigh 500mg of Sodium salicylate and transfer it into 250ml
volumetric flask dissolve and dilute with 96% v/v alcohol.
Stock solution-2:-
Weigh 100mg of Sodium salicylate and transfer it into 250ml
volumetric flask dissolve and dilute with 96% v/v alcohol.
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16. • Procedure:-
Spot 5 micro liter of each solution observe at 254nm and 366nm.
• Acceptance:-
1. The spot shall be comparable intensity wise.
2. Spot due to stock solution-2 shall be visible at 254nm.
3. Spot due to stock solution-1 shall be visible at 366nm.
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17. REFERENCE
1. Textbook of pharmaceutical analysis 4th edition Dr. Ravi Shankar
2. Google source https://www.pharmaguideline.com/2011/09/sop-for-
calibration-of-high-performance.html
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18. THANK YOU
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