CLEANING VALIDATION & ANALYTICA METHOD VALIDATION.

GUIDED BY presented by RENUKA

Dr. K. KISHORE KUMAR 256213886008

Cleaning Validation

Principle• The objectives of GMP include prevention of possible

contamination and cross-contamination

• Contamination by a variety of substances

– contaminants (e.g. microbes, previousproducts (both API and excipient residues),residues of cleaning agents, airbornematerials (e.g. dust and particulate matter),lubricants and ancillary material, such asdisinfectants

• Also decomposition residues from product or detergents

Reference :- [1 ]

Scope• Guidelines: General aspects of cleaning validation

• Excluding specialized cleaning or inactivation

– e.g. for removal of viral or mycoplasmalcontaminants in the biological manufacturingindustry.

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

– between manufacturing of one product andanother

– contact surfaces (products, drug products andAPI).

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 equipmentand 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 ofcleaning agent and rinse solution; and

– the manufacturer's recommendations

Reference :- [3]

Cleaning validation protocols (5)Bracketing:

• Very similar cleaning procedures for products andprocesses - no need for individual validation. “Worst case”may be acceptable and should be justified.

• Consider type of products and equipment; allowed onlywhere products are similar in nature or property andprocessed on the same equipment; and identical cleaningprocedures used.

Reference :- [3 ]

Cleaning validation protocols (6)Bracketing:

• Representative product - most difficult to clean.

• Equipment - only when it is similar or the same equipment indifferent sizes (e.g. 300 l, 500 l and 1000 l tanks).

– Alternative approach may be to validate thesmallest 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 presentedin cleaning validation reports stating the outcome andconclusion.

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 differentproducts.

• The design of equipment may influence the effectiveness of thecleaning 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 - withacceptable 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 andbulk drug or

– where the drying of residues will directlyaffect 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 notvalidated

– may indicate presence of unacceptableresidue and contaminants resulting from anineffective cleaning process.

Reference :- [4 ]

Direct surface sampling (direct method)

• Most commonly used method

• Use “swabs” (inert material) - type of sampling material should notinterfere 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 inthe protocol)

– composition of the equipment (e.g. glass orsteel)

• 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 beroutinely disassembled

• Provides an "overall picture"

• Useful for checking for residues of cleaning agents

• In combination with other sampling methods such as surfacesampling

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 ofcontaminant 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 PROCEDUREFOR THE EQUIPMENT -

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

Reference :- [5 ]

DEVELOP AND VALIDATE THE SAMPLING AND CHOSENANALYTICAL METHODS FOR THE COMPOUND(S) BEINGCLEANED

• 1. Swab 2. Rinse

• (determine % recovery, limit of detection, limit ofquantitation, accuracy of method, reproducibility, stabilityover 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 (rinsesampling)

• 1. Equipment surface area (necessary to calculate carryoverinto subsequent batches)

Reference :- [5 ]

Step-2

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

• That should encompass for example: 1. Introduction 2. Scope3. 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 VALIDATIONREPORTS ON A CLEAN BY CLEAN BASIS DETAILING THEACCEPTABILITY OF THE CLEANING PROCEDURE FOR THEEQUIPMENT AND THE PRODUCT.

• This is only required where there is a long period of timebetween manufacture of the validation runs (see stage 4 forreporting 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. Analyticaltesting procedure 7. Acceptance/Cleaning limits. 8. Acceptance criteria forthe validation.

• GENERATE A CLEANING VALIDATION REPORT DETAILING THEACCEPTABILITY OF THE CLEANING PROCEDURE FOR THE EQUIPMENT ANDTHE PRODUCT

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

Reference :- [5 ]

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 tochemical 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 differentdosage forms based on physiologicalresponse (this method is essential for potentmaterials)

Reference :- [6 ]

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

– Visually clean No residue visible on equipmentafter cleaning. Spiking studies to determine theconcentration at which most active ingredientsare visible. (May not be suitable5 for highpotency, low-dosage drugs.)

– No more than 10 ppm of one product will appearin another product (basis for heavy metals instarting materials).

– No more than 0.1% of the normal therapeuticdose of one product will appear in the maximumdaily dose of a subsequent product.

Reference :- [6 ]

Introduction

• Method validation is the process used to confirm that theanalytical procedure employed for a specific test is suitablefor its intended use. Results from method validation can beused to judge the quality, reliability and consistency ofanalytical results; it is an integral part of any good analyticalpractice.

• 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 methidvalidation

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 whichtest results generated by the method and the true valueagree. Accuracy can also be described as the closeness ofagreement between the value that is adopted, either as aconventional, true or accepted reference value, and the valuefound.

Reference :- [10 ]

Linearity and Calibration Curve

• The linearity of an analytical method is its ability to elicit test results thatare directly proportional to the concentration of analytes in sampleswithin a given range or proportional by means of well-definedmathematical transformations. Linearity may be demonstrated directly onthe test substance (by dilution of a standard stock solution) and/or byusing separate weighings of synthetic mixtures of the test productcomponents, using the proposed procedure.

• Linearity is determined by a series of 3 to 6 injections of 5 or morestandards whose concentrations span 80–120 percent of the expectedconcentration range. The response should be directly proportional to theconcentrations of the analytes or proportional by means of a well-definedmathematical calculation. A linear regression equation applied to theresults should have an intercept not significantly different from 0. If asignificant nonzero intercept is obtained, it should be demonstrated thatthis has no effect on the accuracy of the method.

Reference :- [10 ]

Range

• The range of an analytical method is the interval between theupper and lower levels (including these levels) that have beendemonstrated to be determined with precision, accuracy andlinearity using the method as written. The range is normallyexpressed 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 tobe 80 to 120 percent of the test concentration, and for thedetermination of an impurity, the range to extend from the limit ofquantitation, or from 50 percent of the specification of eachimpurity, 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 valueis larger than the uncertainty associated with it. It is thelowest concentration of analyte in a sample that can bedetected but not necessarily quantified. The limit of detectionis frequently confused with the sensitivity of the method. Thesensitivity of an analytical method is the capability of themethod to discriminate small differences in concentration ormass of the test analyte. In practical terms, sensitivity is theslope of the calibration curve that is obtained by plotting theresponse against the analyte concentration or mass.

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

• Visual inspection: The detection limit is determined by theanalysis of samples with known concentrations of analyte andby establishing the minimum level at which the analyte can bereliably detected.

Reference :- [10 ]

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

Reference :- [10 ]

•Standard deviation of the response based on the standarddeviation of the blank: Measurement of the magnitude ofanalytical background response is performed by analyzingan appropriate number of blank samples and calculating thestandard deviation of these responses.

Ruggedness

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

Reference :- [10 ]

Robustness

• Robustness tests examine the effect that operationalparameters have on the analysis results. For thedetermination of a method’s robustness, a number ofmethod parameters, for example, pH, flow rate, columntemperature, injection volume, detection wavelength ormobile phase composition, are varied within a realisticrange, and the quantitative influence of the variables isdetermined. If the influence of the parameter is within apreviously specified tolerance, the parameter is said tobe 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 PharmaceuticalManufacture - Recommendations on Cleaning Validation. (ref.Document PR 1/ 99 March 1999.

3. Zeller, ’Cleaning Validation and residue limits: a contributionto 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 biologicalproducts: case study’, pharmaceutical engineering.(November / December 1995).

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

7. ISO/IEC 17025.international standad. Eurachem. guidancedocument 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).