This presentation explains about principle involved in HPTLC ,salient features and application of HPTLC ,Qualification of HPTLC

1. QUALIFICATION OF HPTLC
PRESENTED BY
SURIYAPRIYA.K
1st yr. M. PHARM
DEPARTMENT OF PHARMACEUTICAL ANALYSIS
KMCH COLLEGE OF PHARMACY
COIMBATORE

2.  QUALIFICATION:
Qualification is part of validation ; Action of providing and documenting
that equipment or ancillary systems are properly installed , work correctly and
actually lead to the expected results.
The entire qualification consists of four parts;
1.Design qualification (DQ)
2.Installation qualification (IQ)
3.Operational qualification (OQ)
4.performance qualification(PQ)

3. PRINCIPLE
HPTLC have the principle of separation is adsorption.
 The mobile phase solvent flows through because of capillary action.
 The components move according to their affinities towards the adsorbent.
 The component with more affinity towards the stationary phase travels slower.
 The component with lesser affinity towards the stationary phase travels faster.
 Thus the components are separated on a chromatographic plate.

5. SALIENT FEATURES OF HPTLC
• It is simple to learn and operate.
• Accuracy and precision of quantification is high.
• Samples rarely require cleanup.
• Low maintenance cost.

6. APPLICATONS OF HPTLC
 Herbal fingerprinting
 Herbal Analysis – Quantification
 Pharmaceutical Science
 Determination of purity of sample and Identification of compounds
 Identification of adulterants
 Forensic science
 Determination of mercury in water
 Analysis of environmental pollution levels
 Determination of ß-blockers like Metoprolol, Alprenolol, Atenolol

8. QUALIFICATIONS
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.
• CAMAG’s ISO 9001 (DE) / ISO 9001 (EN) certified Quality Management
System ascertains planned testing procedures, error reporting and controlled
updating of documents.

9. To be continued,
• Compliance is documented, e.g. "Declaration of System Validation" and
"Declaration of Conformity" supplied with specific products.
• To make sure the laboratory is equipped with the necessary supporting
equipment etc.

10. Installation Qualification (IQ)
• This 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, approved by CAMAG.

11. Operation Qualification (OQ)
• This qualification is performed subsequent to 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 CAMAG.

12. • Repetitive OQ’s can be performed by a system user well aquatinted with
the system, following guidelines issued by CAMAG.
• On request of the customer, such OQ’s can be performed by a Product
Specialist or Service Engineer approved by CAMAG, against a fee or
within a service contract.
TO BE CONTINUED,

13. 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, the CAMAG OQ procedure, the different
instrument manuals and the customer's QC requirements.

14. Check the performance of HPTLC in terms of
A. Linearity of spotting
B. Reproducibility of Spotting
Requirements for calibration of HPTLC
1. Mobile phase
2. Sample solution
3. HPTLC plates

15. A .LINEARITY OF SPOTTING
• Apply 2 µl, 4µl, 6µl, 8µl and 10 µl of solution on HPTLC plate with spotter.
• Allow the plate to run in mobile phase.
• Dry the plate with dryer.
• Scan the plate with scanner.
• Check the linearity and correlation coefficient between the spots.

16. Calculation of correlation coefficient:
It is a number that gives you a good idea about how closely one variable
(spot) is related to another variable.
Acceptance limit
Correlation coefficient is not less than 0.9900

17. 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.
 RSD Limit: not more than 3.0%

18. REFERENCE
 Ph.Eur.2.2.35 chromatography; High performance liquid
chromatography
 Guidance on equipment qualification of analytical instruments
 Journal of Perkin Elmer life & analytical science
 https://www.slideshare.net/huzaifanaaz/qualification-of-high-
performance-liquid-chromatography