CLEANING VALIDATION & ANALYTICA METHOD VALIDATION.

GUIDED BY presented by RENUKADr. K. KISHORE KUMAR 256213886008

Cleaning Validation

Cleaning ValidationPrinciple• The objectives of GMP include prevention of possible

contamination and cross-contamination• Contamination by a variety of substances

– contaminants (e.g. microbes, previous products (both API and excipient residues), residues of cleaning agents, airborne materials (e.g. dust and particulate matter), lubricants and ancillary material, such as disinfectants

• Also decomposition residues from product or detergentsReference :- [1 ]

Scope• Guidelines: General aspects of cleaning validation• Excluding specialized cleaning or inactivation

– e.g. for removal of viral or mycoplasmal contaminants in the biological manufacturing industry.

• Normally cleaning validation needed for critical cleaning, e.g.

– between manufacturing of one product and another

– contact surfaces (products, drug products and API).

Reference :- [1 ]

Cleaning validation protocols (1)• Approved by QC or QA and to cover, e.g.

– disassembly of system;– pre-cleaning;– cleaning agent, concentration, solution volume,

water quality;– time and temperature;– flow rate, pressure and rinsing;– complexity and design of the equipment;– training of operators; and– size of the system.

Reference :- [2 ]

Cleaning validation protocols (2)• The cleaning validation protocol should include:

– objectives, responsible people; – description of the equipment including the make,

model, serial number or other unique code; – time intervals; bioburden; 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 .

Reference :- [3]

Cleaning validation protocols (3)• The cleaning validation protocol should include (2):

– data on recovery studies (efficiency of the recovery of the sampling technique should be established);

– analytical methods; – acceptance criteria (with rationale for setting the

specific limits) including a margin for error and for sampling efficiency;

– cleaning agent to be used;– revalidation requirements.

Reference :- [3]

Cleaning validation protocols (4)• Cleaning agent used, scientifically justified and based on:

– the solubility of the materials to be 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 recommendations

Reference :- [3]

Cleaning validation protocols (5)Bracketing:• Very similar cleaning procedures for products and

processes - no need for individual validation. “Worst case” may be acceptable and should be justified.

• Consider type of products and equipment; allowed only where products are similar in nature or property and processed on the same equipment; and identical cleaning procedures used.

Reference :- [3 ]

Cleaning validation protocols (6)Bracketing:• Representative product - most difficult to clean.• Equipment - only when it is similar or the same equipment in

different sizes (e.g. 300 l, 500 l and 1000 l tanks).

– Alternative approach may be to validate the smallest and the largest sizes separately.

Reference :- [3]

Cleaning validation reports• The relevant cleaning records – (signed by the operator,

checked by production and reviewed by quality assurance) – and source data (original results) should be kept.

• The results of the cleaning validation should be presented in cleaning validation reports stating the outcome and conclusion.

Reference :- [4 ]

Equipment • If one SOP for cleaning a piece of equipment, review:

– products being produced, – cleaning in a large campaign, – cleaning between batches of different

products.• The design of equipment may influence the effectiveness of the

cleaning process.• Consider design, e.g. V-blenders, transfer pumps or filling lines.

Reference :- [5 ]

Detergents• Released by quality control and meet food standards or

regulations• Composition known• Easily removed with rinsing - demonstrated - with

acceptable limits defined• If persistent residues (e.g. cationic detergents) - avoided• Consider also detergent breakdown

Reference :- [3 ]

Microbiology

• Prevent microbial growth and remove contamination• Documented evidence

– routine cleaning– storage of equipment

• The period and conditions

– storage of unclean equipment before cleaning– between cleaning and equipment reuse

• Equipment stored in a dry condition after cleaning (no stagnant water)

• Control of bioburden important

Reference :- [5 ]

Sampling• Clean as soon as possible after use

– especially topical products, suspensions and bulk drug or

– where the drying of residues will directly affect the efficiency of a cleaning procedure

• Two methods of sampling:

– direct surface sampling and – rinse samples

• Combination of the two - most desirable

Reference :- [4 ]

Sampling

• Re-sampling:

– not to be done before or during cleaning• Constant re-testing and re-sampling:

– can show that the cleaning process is not validated

– may indicate presence of unacceptable residue and contaminants resulting from an ineffective cleaning process.

Reference :- [4 ]

Direct surface sampling (direct method)• Most commonly used method• Use “swabs” (inert material) - type of sampling material should not

interfere with the test• Factors to be considered include:

– supplier of the swab,– area swabbed, number of swabs used,

whether they are wet or dry swabs,– swab handling and swabbing technique

Reference :- [4 ]

Direct surface sampling (direct method) (2)• Other factors include:

– location from which the sample is taken (including worst case locations, identified in the protocol)

– composition of the equipment (e.g. glass or steel)

• Critical areas (hardest to clean)

– e.g. in semi-automatic/fully automatic clean-in-place systems

• Use appropriate sampling medium and solvent

Reference :- [4 ]

Rinse samples (indirect method)• Allows sampling of:

– a large surface – areas that are inaccessible or that cannot be

routinely disassembled• Provides an "overall picture"• Useful for checking for residues of cleaning agents• In combination with other sampling methods such as surface

sampling

Reference :- [4 ]

Rinse samples (indirect method) (2)

The manufacturer has to provide evidence that samples are accurately recovered

What is considered acceptable in terms of recovery?

Reference :- [4 ]

Levels of cleaning• LEVEL USED WHEN CLEANING VALIDATION REQUIRED• LEVEL 2 • Product changeover of equipment used in final step • Intermediates of one batch to final step of another• LEVEL 1 • Intermediates or final Step of one product to intermediate of another • Early Step to intermediates in a product sequence• progression between level 0 and 2 depending on process and nature of

contaminant based on scientific rational• LEVEL 0- in-campaign, batch to batch changeover• no validation required• NB: ALL PROCESSES MUST BE EVALUATED INDIVIDUALLY

Reference :- [5 ]

Cleaning validation processstep-1

DETERMINE THE MOST APPROPRIATE CLEANING PROCEDURE FOR THE EQUIPMENT -

• 1. Generate acceptance criteria data for the contaminant. 2. The cleaning method will be determined by the process, the equipment the cleaning agents and the cleaning techniques available. 3. All aspects of the cleaning procedure should be clearly defined in SOPs be they manual / CIP or COP

Reference :- [5 ]

DEVELOP AND VALIDATE THE SAMPLING AND CHOSEN ANALYTICAL METHODS FOR THE COMPOUND(S) BEING CLEANED

• 1. Swab 2. Rinse• (determine % recovery, limit of detection, limit of

quantitation, accuracy of method, reproducibility, stability over time ...etc.)

Reference :- [5 ]

EVALUATE EQUIPMENT SURFACES AND DETERMINE• 1. Worst case locations to sample (swab sampling)• 1. Volume and type of rinse solvent to be employed (rinse

sampling)• 1. Equipment surface area (necessary to calculate carryover

into subsequent batches)

Reference :- [5 ]

Step-2

DEVELOP A CLEANING VALIDATION PROTOCOL FOR THE PRODUCT AND THE EQUIPMENT BEING CLEANED

• That should encompass for example: 1. Introduction 2. Scope 3. Equipment 4. Cleaning procedure 5. Sampling procedures 6. Analytical testing procedure 7. Acceptance/Cleaning limits. 8. Acceptance criteria for the validation.

Reference :- [5 ]

Step-3

INTERIM REPORT: GENERATE INTERIM CLEANING VALIDATION REPORTS ON A CLEAN BY CLEAN BASIS DETAILING THE ACCEPTABILITY OF THE CLEANING PROCEDURE FOR THE EQUIPMENT AND THE PRODUCT.

• This is only required where there is a long period of time between manufacture of the validation runs (see stage 4 for reporting requirements).

Reference :- [5 ]

Step-4 FOR THE PRODUCT AND THE EQUIPMENT BEING CLEANED• That should encompass for example: 1. Introduction 2. Scope 3.

Equipment 4. Cleaning procedure 5. Sampling procedures 6. Analytical testing procedure 7. Acceptance/Cleaning limits. 8. Acceptance criteria for the validation.

• GENERATE A CLEANING VALIDATION REPORT DETAILING THE ACCEPTABILITY OF THE CLEANING PROCEDURE FOR THE EQUIPMENT AND THE PRODUCT

• The report should give a full detailed background and introduction to the cleaning Validation study and should evaluate all data generated with respect to the acceptance criteria employed for the study. The report should also indicate the requirement if any for revalidation (period of time /change control etc.)

Reference :- [5 ]

Validation

Recovery

> 80% is considered good

Reference :- [6 ]

Validation

Recovery

> 80% is considered good > 50% is considered

reasonable

Reference :- [6 ]

Validation

Recovery

< 50% is consideredquestionable

> 80% is considered good > 50% is considered

reasonable

Reference :- [6 ]

Establishing acceptable limits

• Limits: Practical, achievable and verifiable• Rationale: Logical, based on knowledge of materials• Each situation assessed individually• Principal reactant and other chemical variations• Screening (thin-layer chromatography) in addition to

chemical analyses where necessary

Reference :- [6 ]

Establishing acceptable limits (2)

There should be no residue from:• Previous product• Reaction by-products and degradants• Cleaning process itself (e.g. detergents or solvents)

Reference :- [6 ]

Establishing acceptable limits (3) The limit-setting approach can:

– be product-specific– group products into families and choose a

worst case product– group products into groups according to risk,

e.g. very soluble products, products with similar potency, highly toxic, or difficult to detect products

– use different safety factors for different dosage forms based on physiological response (this method is essential for potent materials)

Reference :- [6 ]

Establishing acceptable limits (4) The three most commonly used criteria are:

– Visually clean No residue visible on equipment after cleaning. Spiking studies to determine the concentration at which most active ingredients are visible. (May not be suitable5 for high potency, low-dosage drugs.)

– No more than 10 ppm of one product will appear in another product (basis for heavy metals in starting materials).

– No more than 0.1% of the normal therapeutic dose of one product will appear in the maximum daily dose of a subsequent product.

Reference :- [6 ]

Analytical Method Validation

Introduction

• 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.

• Analytical methods need to be validated or revalidated

Reference :- [7 ]

Once the method has been developed and validated, a validation report should be prepared that includes the following:• Objective and scope of the method (applicability, type).• Summary of methodology.• Type of compounds and matrix.• All chemicals, reagents, reference standards, QC samples with purity, grade, their source or detailed instructions on their preparation. • Procedures for quality checks of standards and chemicals used.• Safety precautions.• A plan and procedure for method implementation from the method development lab to routine analysis.• Method parameters.• Critical parameters taken from robustness testing.• Listing of equipment and its functional and performance requirements, e.g., cell dimensions, baseline noise and column temperature range. For complex equipment, a picture or schematic diagram may be useful.

Reference :- [7 ]

• Detailed conditions on how the experiments were conducted, including sample preparation. The report must be detailed enough to ensure that it can be reproduced by a competent technician with comparable equipment. • Statistical procedures and representative calculations.• Procedures for QC in routine analyses, e.g., system suitability tests.• Representative plots, e.g., chromatograms, spectra and calibration curves.• Method acceptance limit performance data.•The expected uncertainty of measurement results.•Criteria for revalidation.•The person(s) who developed and validated the method.•References (if any).•Summary and conclusions.•Approval with names, titles, date and signature of those responsible for the review and approval of the analytical test procedure.

Reference :- [8]

Parameters for analytical methid validation

Reference :- [8 ]

Selectivity $ specificity• It is a method that produces a response for a single analyte

only, while the term selective refers to a method that provides responses for a number of chemical entities that may or may not be distinguished from each other. If the response is distinguished from all other responses, the method is said to be selective

• Selectivity studies should also assess interferences that may be caused by the matrix, e.g., urine, blood, soil, water or food. Optimized sample preparation can eliminate most of the matrix components. The absence of matrix interferences for a quantitative method should be demonstrated by the analysis of at least five independent sources of control matrix.

Reference :- [8 ]

Precision and Reproducibility

• The precision of a method (Table 4) is the extent to which the individual test results of multiple injections of a series of standards agree. The measured standard deviation can be subdivided into 3 categories: repeatability, intermediate precision and reproducibility (4, 5). Repeatability is obtained when the analysis is carried out in a laboratory by an operator using a piece of equipment over a relatively short time span. At least 6 determinations of 3 different matrices at 2 or 3 different concentrations should be performed, and the RSD calculated.

Reference :- [9 ]

Accuracy and Recovery

• The accuracy of an analytical method is the extent to which test results generated by the method and the true value agree. Accuracy can also be described as the closeness of agreement between the value that is adopted, either as a conventional, true or accepted reference value, and the value found.

Reference :- [10 ]

Linearity and Calibration Curve

• The linearity of an analytical method is its ability to elicit test results that are directly proportional to the concentration of analytes in samples within a given range or proportional by means of well-defined mathematical transformations. Linearity may be demonstrated directly on the test substance (by dilution of a standard stock solution) and/or by using separate weighings of synthetic mixtures of the test product components, using the proposed procedure.

• Linearity is determined by a series of 3 to 6 injections of 5 or more standards

whose concentrations span 80–120 percent of the expected concentration range. The response should be directly proportional to the concentrations of the analytes or proportional by means of a well-defined mathematical calculation. A linear regression equation applied to the results should have an intercept not significantly different from 0. If a significant nonzero intercept is obtained, it should be demonstrated that this has no effect on the accuracy of the method.

Reference :- [10 ]

Range

• The range of an analytical method is the interval between the upper

and lower levels (including these levels) that have been demonstrated to be determined with precision, accuracy and linearity using the method as written. The range is normally expressed in the same units as the test results (e.g., percentage, parts per million) obtained by the analytical method.

• For assay tests, the ICH (5) requires the minimum specified range to be 80 to 120 percent of the test concentration, and for the determination of an impurity, the range to extend from the limit of quantitation, or from 50 percent of the specification of each impurity, whichever is greater, to 120 percent of the specification.Reference :- [10 ]

Limit of Detection

• The limit of detection is the point at which a measured value is larger than the uncertainty associated with it. It is the lowest concentration of analyte in a sample that can be detected but not necessarily quantified. The limit of detection is frequently confused with the sensitivity of the method. The sensitivity of an analytical method is the capability of the method to discriminate small differences in concentration or mass of the test analyte. In practical terms, sensitivity is the slope of the calibration curve that is obtained by plotting the response against the analyte concentration or mass.

• In chromatography, the detection limit is the injected amount that results in a peak with a height at least two or three times as high as the baseline noise level. Besides this signal/noise method, the ICH (4) describes three more methods:

• Visual inspection: The detection limit is determined by the analysis of samples with known concentrations of analyte and by establishing the minimum level at which the analyte can be reliably detected.

Reference :- [10 ]

•Standard deviation of the response based on the slope of the calibration curve: A specific calibration curve is studied using samples containing an analyte in the range of the limit of detection. The residual standard deviation of a regression line, or the standard deviation of y-intercepts of regression lines, may be used as the standard deviation.

Reference :- [10 ]

•Standard deviation of the response based on the standard deviation of the blank: Measurement of the magnitude of analytical background response is performed by analyzing an appropriate number of blank samples and calculating the standard deviation of these responses.

Ruggedness

• Ruggedness is a measure of reproducibility of test results under normal, expected operational conditions from laboratory to laboratory and from analyst to analyst. Ruggedness is determined by the analysis of aliquots from homogeneous lots in different laboratories.

Reference :- [10 ]

Robustness

• Robustness tests examine the effect that operational parameters have on the analysis results. For the determination of a method’s robustness, a number of method parameters, for example, pH, flow rate, column temperature, injection volume, detection wavelength or mobile phase composition, are varied within a realistic range, and the quantitative influence of the variables is determined. If the influence of the parameter is within a previously specified tolerance, the parameter is said to be within the method’s robustness range.

Reference :- [8 ]

References1. ICH Good Manufacturing Practice Guideline for Active

Pharmaceutical Ingredients. (July 23 1999).2. Principles of Qualification and Validation in Pharmaceutical

Manufacture - Recommendations on Cleaning Validation. (ref. Document PR 1/ 99 March 1999.

3. Zeller, ’Cleaning Validation and residue limits: a contribution to current discussions’, pharmaceutical technology Europe. (November 1993).

4. S.W. Harder, ‘The validation of cleaning processes’, pharmaceutical technology. (1984)

5. Mc Arthur, Vasilevsky, ‘Cleaning validation for biological products: case study’, pharmaceutical engineering. (November / December 1995).

6. James Agalloco, ‘Points to consider in the validation of equipment cleaning procedures’, Journal of parenteral science and technology. (October 1992).

7. ISO/IEC 17025.international standad. Eurachem. guidance document no.1/WELAC guidance document no.WGD 2.

8. ICH of technical requirements for the regestration of pharmaceuticals for human use.

9. Green, J.M.A practical guide to analytical method validation.

10. Eurachem. guidance document no.1/WELAC guidance document no.WGD 2:accreditation for chemical lads & ISO/IEC guide 25 (1993).