RIPE

ANALYTICAL METHOD R
AUTONOM

VALIDATION USED IN CLEANING OUS

NAAC &
NBA(UG)
A seminar presented as a part of SIRO - DSIR

1st Year M. Pharmacy 2nd semester requirement

ACY 2019-2020
By
T. Reshma
M. Pharmacy 1 – 2 semester
Reg. No: 19L81S0407
Pharmaceutical Quality Assurance

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CONTENTS R
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1.Cleaning validation NAAC &
NBA(UG)
2.Protocols SIRO - DSIR

3.Levels of Cleaning
4.Process of Cleaning validation
5.Analytical method validation
6.What is validation
7.Objectives of validation
8.Parameters assessed during analytical validation
9.References

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R
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NAAC &
NBA(UG)
SIRO - DSIR

CLEANING
VALIDATIO
N

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Why cleaning validation is so important AUTONOM
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NAAC &
NBA(UG)
Pharmaceuticals can be contaminated by SIRO - DSIR

potentially dangerous substances.

Particular attention should be accorded to the
validation of cleaning procedures.
Cleaning validation should be performed in
order to confirm the effectiveness of a cleaning
procedure.

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Possible Contaminants R
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• Product residues OUS
NAAC &
NBA(UG)
• Cleaning agent residues and breakdown SIRO - DSIR

• Airborne matter

• Lubricants, ancillary material

• Decomposition residues

• Bacteria, mould and pyrogens

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CLEANING VALIDATION PROTOCOLS R
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1. Approved by QC or QA and to cover, e.g. NAAC &
NBA(UG)
SIRO - DSIR

– Disassembly of system
– Pre-cleaning
– Cleaning agent
– Time and Temperature
– Flow rate, Pressure and Rinsing
– Complexity and Design of the equipment
– Training of operators and
– Size of the system.

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2. The cleaning validation protocol: R
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– Objectives, responsible people NAAC &
NBA(UG)
– Description of the equipment including the model, SIRO - DSIR
serial number or other unique code
– Time intervals, cleaning procedures
– Equipment used for routine monitoring (e.g.
conductivity meters, pH meters and total organic carbon
analysers)
– Number of cleaning cycles; sampling procedures
(e.g. direct sampling, rinse sampling, in process
monitoring and sampling locations) and the rationale for
their use .

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3. The cleaning validation protocol R
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– Data on recovery studies OUS
NAAC &
NBA(UG)
SIRO - DSIR
– Analytical methods

– Acceptance criteria

– Cleaning agent to be used

– Revalidation requirements.

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4. Cleaning agent used: AUTONOM
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NAAC &
– The solubility of the materials to be NBA(UG)
SIRO - DSIR
removed
– The design and construction of the
equipment and surface materials to be cleaned
– The safety of the cleaning agent
– The ease of removal and detection
– The product attributes
– The minimum temperature and volume of
cleaning agent and rinse solution and
– The manufacturer's recommendation

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LEVELS OF CLEANING R
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There are 4 levels of cleaning, they are: NAAC &
NBA(UG)
A.Level 1 SIRO - DSIR

B.Level 2
C.Level 3
D.Level 4

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Level 1 cleaning: R
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NAAC &
It is used only between steps in the same NBA(UG)
manufacturing process SIRO - DSIR

Level 2 cleaning:
Level 2 cleaning would be used if step B was to
be performed immediately after step A for the
same product line.

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Level 3 cleaning: R
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NAAC &
It would be performed when cleaning after an NBA(UG)
intermediate or final product step or one product SIRO - DSIR

in preparation of an intermediate step of another

product.

Level 4 cleaning:
It would be used after final product is ready.

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 Level 1 & Level 2 cleaning : AUTONOM
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NAAC &
• Lowest risk NBA(UG)
SIRO - DSIR
• Higher limits
• Less extensive cleaning
• Visual verification of clean

 Level 3 & Level 4 cleaning:

• Highest risk
• Lower limits
• More extensive cleaning
• Analytical method

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The cleaning process validation takes the R
following into account: AUTONOM
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NAAC &
 Validation of Cleaning Processes NBA(UG)
SIRO - DSIR

 Equipment and Personnel

 Microbiological Considerations

 Documentation

 Sampling, Rinsing, Rinse Samples and Detergents

 Establishment of Limits

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VALIDATION OF CLEANING PROCESSES R
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This concept involves cleaning, sampling, and OUS
NAAC &
testing with repetition of this sequence until an NBA(UG)
SIRO - DSIR
acceptable residue limit is attained.

If the automated procedures are utilized, consideration

should be given to monitoring the critical control points

During process, cleaning between the batches may be

reduced.

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EQUIPMENT AND PERSONNEL R
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All processing equipment should be specifically OUS
NAAC &
designed to facilitate cleanability and permit NBA(UG)
SIRO - DSIR
visual inspection and whenever possible, the
equipment should be made of smooth surfaces
of non-reactive materials.
It is difficult to validate a manual cleaning procedure (i.e.
an inherently variable/cleaning procedure). Therefore,
operators carrying out manual cleaning procedures should
be adequately trained, monitored, and periodically
assessed.

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MICROBIOLOGICAL CONSIDERATIONS R
The existence of conditions favourable to reproduction
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of micro-organisms (e.g. moisture, temperature, NAACand&
NBA(UG)
rough surfaces) and the time of storage should be
SIRO - DSIR
considered.
Equipment should be dried before storage, and under no
circumstances should stagnant water be allowed to remain
in equipment subsequent to cleaning operations.
Conditions of storage of equipment before cleaning and
the time between cleaning and equipment reuse is a part of
the validation of cleaning procedures.
This is to provide confidence that does not allow
microbial proliferation.

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DOCUMENTATION R
Detailed cleaning procedure(s) are to be AUTONOM OUS
documented in SOPs NAAC &
NBA(UG)
 A cleaning validation protocol should include the SIRO - DSIR
following:
• The objective of the validation process.
• Responsibilities for performing and approving the
validation study.
• Description of the equipment to be used.
• The interval between the end of production and the
beginning of the cleaning procedure.
• Detailed cleaning procedures to be used for each product.

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• Sampling procedures, including the rationale for R
why a certain sampling method is used. AUTONOM
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• Clearly defined sampling locations. NAAC &
NBA(UG)
SIRO - DSIR

• The number of cleaning cycles to be performed

consecutively.

• A Final Validation Report should be prepared. The

conclusions of this report should state that cleaning
process has been validated successfully.

• The report should be approved by the Plant

Management.
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SAMPLING, RINSING, RINSE SAMPLES R
AND DETERGENTS AUTONOM
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NAAC &
Sampling: NBA(UG)
SIRO - DSIR

There are two general types of sampling that are

considered to be acceptable they are:
1. Direct surface sampling (swab method)
2. Indirect sampling (use of rinse
solutions)
 A combination of the two methods is generally the most
desirable.

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Detergents: R
Detergents should be easily removable, being AUTONOM
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used to facilitate the cleaning during the NAAC &
NBA(UG)
cleaning process. SIRO - DSIR

When detergents are used in the cleaning process, their

composition should be known to the user and their
removal should be demonstrated.

Acceptable limits should be defined for detergent

residues after cleaning

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Last Rinse: R
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Water for injection should be used as the last NAAC &
NBA(UG)
rinse for product-contact equipment to be SIRO - DSIR
utilized in the fabrication of sterile products.

Purified water is considered acceptable as the last rinse

for product-contact equipment used in the fabrication of
non-sterile products or sterile products for ophthalmic
use.

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ESTABLISHMENT OF LIMITS NAAC &
NBA(UG)
SIRO - DSIR

The pharmaceutical company's rationale for selecting

limits for product residues should be logically based on a
consideration of the materials involved and their
therapeutic dose. The limits should be practical,
achievable and verifiable.

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R
AUTONOM
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NAAC &
NBA(UG)
SIRO - DSIR

ANALYTICAL
METHOD
VALIDATION

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VALIDATION R
AUTONOM
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NAAC &
Validation is defined as the “Finding or testing NBA(UG)
SIRO - DSIR
the truth of some thing.”

Method validation is the process used to confirm that the

analytical procedure employed for a specific test is
suitable for its intended use.

Results from method validation can be used to judge the

quality, reliability and consistency of analytical results, it
is an integral part of any good analytical practice.

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OBJECTIVES R
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• It reduces risk of regulatory non-compliance. NAAC &
NBA(UG)
SIRO - DSIR
• Reduction of time to the market for the new products.

• Eliminates the scrap & reduces the defect cost.

• Reduces the chances of product re-call from market.

• It requires less in-process control & end process testing.

• Parametric release of batch can be achieved in validation.

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Parameters evaluate during analytical R
AUTONOM
method validation: OUS
NAAC &
NBA(UG)
SIRO - DSIR
1. Linearity
2. Range
3. Specificity
4. Precision
5. Accuracy
6. Limit of Detection
7. Limit of Quantitation
8. Robustness
9. System suitability

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Linearity R
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Linearity is the ability of a method, to measure the OUS
NAAC &
analyte accurately in the presence of all potential NBA(UG)
sample components. SIRO - DSIR

For establishment of linearity,

minimum 5 concentrations are
recommended.

Linearity results should be

established by appropriate
statistical methods.

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LINEARITY FOR CONC vs RESPONSE R
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NAAC &
NBA(UG)
SIRO - DSIR

The following parameters should be determined:

• correlation coefficient
• y-intercept
• slope of the regression line

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R
AUTONOM
Range OUS
NAAC &
NBA(UG)
SIRO - DSIR
 The range of the procedure is validated by verifying that
the analytical procedure provides acceptable precision,
accuracy, and linearity when applied to samples
containing analyte at the extremes of the range as well
as within the range.

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Specificity R
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The ability to detect the analyte in the presence of OUS
NAAC &
interfering substances like impurities, degradants and NBA(UG)
matrix is called as specificity. SIRO - DSIR

Identification:
• Suitable identification tests should be able to discriminate between
compounds of closely related structures which are likely to be
present.
• The discrimination of a procedure may be confirmed by obtaining
positive results from samples containing the analyte.
• The identification test may be applied to materials structurally
similar to or closely related to the analyte to confirm that a positive
response is not obtained.

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Assay and impurity test: R
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A. Impurities are available: OUS
 For the assay, this should involve demonstration of the NAAC &
NBA(UG)
discrimination of the analyte in the presence of impurities SIRO - DSIR
and/or excipients.
 This can be done by spiking pure substances with
appropriate levels of impurities and/or excipients and
demonstrating that the assay result is unaffected by the
presence of these materials.
 For the impurity test, the discrimination may be established
by spiking drug substance or drug product with appropriate
levels of impurities and demonstrating the separation of these
impurities individually and/or from other components in the
sample matrix.
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B. Impurities are not available: R
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 If impurity or degradation product standards are NAAC &
unavailable, specificity may be demonstrated by NBA(UG)
SIRO - DSIR
comparing the test results of
samples containing impurities or degradation products to
a second well characterized procedure
e.g. pharmacopeial method or other validated
analytical procedure.
 As appropriate, this should include samples stored under
relevant stress conditions: light, heat, humidity, acid/base
hydrolysis and oxidation

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Precision R
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 The precision of an analytical procedure OUS
NAAC &
expresses the closeness of agreement (degree NBA(UG)
of scatter) between a series of measurements SIRO - DSIR

obtained from multiple sampling of the same

homogeneous sample under the prescribed
conditions.
 Less variation in the absorbance shows High precision

 In the same way High variation in the absorbance

shows Less precision

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R
AUTONOM
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NAAC &
NBA(UG)
SIRO - DSIR

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 Precision may be considered at three levels: R
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Precision
NAAC &
NBA(UG)
SIRO - DSIR

Repeatability

Intermediate
precision
Reproducibility

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Repeatability: R
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Repeatability expresses the precision under the OUS
NAAC &
same operating conditions over a short interval NBA(UG)
SIRO - DSIR
of time.
Repeatability should be assessed using a minimum of 9
determinations covering the specified range.
Intermediate Precision:
Intermediate precision expresses variations within
laboratories, such as different days, different analysts,
different equipment, and so forth

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R
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Reproducibility: NAAC &
NBA(UG)
SIRO - DSIR
Reproducibility expresses the precision between
laboratories. It is assessed by means of an interlaboratory
trial.

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Accuracy R
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 The closeness of agreement between the OUS
NAAC &
conventional true value or an accepted reference NBA(UG)
value and the value found. SIRO - DSIR

 High accuracy shows Less precision and Less accuracy

shows High precision.
Evaluation of Accuracy:
 Accuracy should be assessed using a minimum of 9

determinations over a minimum of 3 concentration levels

covering the specified range
Accuracy should be reported as percent as the difference
between the mean and the accepted true value.
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R
AUTONOM
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NAAC &
NBA(UG)
SIRO - DSIR

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Limit of Detection R
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It is the lowest amount of analyte in a sample OUS
NAAC &
which can be detected but not necessarily NBA(UG)
SIRO - DSIR
quantitated.

Limit of Quantitation
It is the lowest amount of analyte in a sample which can
be quantitatively determined with suitable precision and
accuracy.

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Determination of LOD & LOQ AUTONOM
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NAAC &
NBA(UG)
Limit of Detection Limit of Quantification SIRO - DSIR

 Based on visual evaluation  Based on visual evaluation

 Based on standard deviation  Based on standard deviation

of response and slope of response and slope

LOD = 3.3 σ / Slope LOD = 10 σ / Slope

 Signal to noise ratio 2:1 or 3:1  Signal to noise ratio 10:1

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Robustness R
AUTONOM
The robustness of an analytical procedure is a measure OUS
NAAC &
of its capacity to remain unaffected by small changes in NBA(UG)
the parameters of a method. SIRO - DSIR

But, deliberate variations in method parameters and

provides an indication of its reliability during normal
usage.
If measurements are susceptible to variations in analytical
conditions, the analytical conditions should be suitably
controlled or a precautionary statement should be included
in the procedure, such as:
• Use solution within 24 hours

• Maintain temperature below 25 degrees

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R
AUTONOM
System Suitability OUS
NAAC &
NBA(UG)
SIRO - DSIR
System suitability testing is an integral part of many
analytical procedures. The tests are based on the concept
that the equipment, electronics, analytical operations and
samples to be analysed constitute an integral system that
can be evaluated as such.

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REFERENCES R
AUTONOM
1. S.W. Harder, "The Validation of Cleaning OUS
NAAC &
Procedures," Pharm. Technol. 8 (5), 29-34 (1984) NBA(UG)
SIRO - DSIR
2. J. Rodehamel, "Cleaning and Maintenance," Pgs 82-87,
University of Wisconsin's Control Procedures in Drug
Production Seminar, July 17-22, 1966, William Blockstein,
Editor, Published by the University of Wisconsin, L.O.C.#66-
64234.
3. Rodehamel, "Cleaning and Maintenance," Pgs 82-87,
University of Wisconsin's Control Procedures in Drug
Production Seminar, July 17-22, 1966, William Blockstein,
Editor, Published by the University of Wisconsin, L.O.C.#66-
64234.
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R
AUTONOM
OUS
NAAC &
NBA(UG)
SIRO - DSIR

THANK
YOU…….!

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