Title: Procedure For Cleaning Validation
Purpose
To describe procedures to be followed during the cleaning validation of processes and
equipments.
Scope
This procedure is to be followed by personnel involved in cleaning validation activities, typically
the following departments are involved, Validation, Quality Assurance, Laboratory, Project
Engineering and Operations.
Definitions
Cleaning Validation: cleaning validation is a validation program to verify that the
processes and procedures used to clean product residue from process equipment and
components, will consistently and significantly reduce the amount of active and/or
excipient(s) and cleaning agent(s) to a concentration within calculated acceptance limits
Actives: The main or primary ingredient in a product, which is intended to induce a change
in the recipient
Acceptance Limit: The quantity of compound(s) permitted to carry-over in subsequent
product without any adverse effects to the product or consumer.
Excipient(s): The ingredients in a product, other than the Active ingredient
Hot Spot: A difficult-to-clean location, which if improperly cleaned would lead to
contamination, which would be uniformly distributed in subsequent batches
(e.g. inside the bulk tank).
Limit of Detection: Limit Of Detection (LOD) represents the lowest detectable mass or
concentration that can be observed by an analytical method.
Limit of Quantitation: The Limit Of Quantitation (LOQ) represents the lowest quantifiable
mass or concentration that can be reliably measured repeatedly and accurately.
NOEL: No Observable Effect Limit. This is the limit at which no effects to the subsequent
product or recipient can be detected.
Residual Carry-Over: The concentration of primary or secondary ingredients from the
previous lot or batch, of solid or liquid product, which is transferred to the current
manufactured lot or batch.
Safety Factor: A value incorporated into the allowable carry-over that ensures the
residual concentration of the Active or excipient ingredients in subsequent batches has
absolutely no effect on consumers or the product, (e.g. 1/10, 1/100, 1/1000, 1/10000, etc.).
Title: Procedure For Cleaning Validation
Scientific Rationale: Theoretical justification for allowing an established concentration of
carry-over into subsequent batches.
Swab: A small utensil with an inert absorbent material at one end used for sampling the pre-
determined surface area of equipment. The material must not affect the sample
Toxicity: A value representing the amount of a chemical substance which causes death in
50% of a test population, commonly referred to as an LD50 value.
The lower the toxicity or LD50 value the more toxic is the drug or chemical
Unadulterated Sample A swab or rinse water sample, which has not been corrupted by
outside influences from materials or mishandling by Operators
Worst Case Product: The product, which presents the greatest challenge to the cleaning
process and the highest probability of potential adverse affects on subsequent production
batches or recipients of the unintended substance.
TOC Total Organic Carbon: TOC Analysis Performed by a Total Organic Carbon
analyser, which quantify the amount of Total Organic Carbon present in a given aqueous
sample
1. Responsibilities
1.1. Departments
The responsibility for validation activities is shared between members of a multi-disciplinary
group as identified in the Validation Plan for a particular project.
1.1.1. Validation
This area is responsible for training in the use of this SOP, document preparation and testing
requirements for particular projects. Provide instruction on specific test procedures and written
test protocols. Develop and implement relevant testing templates and calculation spreadsheets.
For large validation projects provide a testing and documentation resource to complete the
validation activities.
1.1.2. Quality Assurance (QA Manager or Validation Manager)
Review and authorisation of documentation associated with cleaning validation.
1.1.3. Engineering (Projects)
Review and checking documentation associated with cleaning validation. Engineering is
responsible for design, installation, and commission and in some projects validation of new and
modified cleaning equipment processes. Systems include but are not limited to: product transfer
Title: Procedure For Cleaning Validation
pipework, mixing vessels. Initiating changes to current cleaning processes and procedures by
initiation of
change requests.
1.1.4. Operations
Initiating changes to current cleaning processes and procedures by initiation of change requests.
Review of validation plans and validation test protocols. Provide resource assistance to the
specific cleaning validation tasks such as running collecting swab and rinse samples, removal of
complex equipment components.
1.1.5. Laboratory
Provide validated Analytical test methods for accurate product residue detection, including swab
and rinse surface recovery data. Perform Analytical testing of swab and rinse samples collected
during validation using validated procedures. Review and approve Analytical test methods and
results, provide documented test results to relevant departments. Review sampling procedures
and acceptance criteria for bioburden sampling after cleaning. Perform analysis of bioburden
samples and report results
2. Types of Cleaning Process and Cleaning Agents
2.1. Manual Cleaning
Effective manual cleaning practices must be established by focusing on the following
two areas:
2.1.1. Standard Operating Procedures (SOP)
SOPs will be developed during the Operational Qualification phase of the project. This
will be outlined as part of the Validation Plan. If consistently unacceptable or erratic
results are obtained the SOP should be considered one of the possible problems and
modifications to the procedure may be required. Procedures must be written in a manner,
which prevents variation between operators.
2.1.2. Operator Training
Operators must be suitably trained in the use of the manual cleaning SOP.
2.2. Automatic Cleaning In Place (CIP)
CIP is the procedure by which flush and rinse solutions are brought into immediate
contact with all internal product soiled surfaces, without the intervention of operators,
and continuously re-circulated or flushing to drain for a pre-determined time. For an
automated procedure the SOP does not need to state precisely how the equipment is
Title: Procedure For Cleaning Validation
cleaned, however validation must document and verify that the automated process
functions according to its design requirements.
CIP Operating Criteria
1. Most CIP applications will use fresh Distilled Water discharged through fixed spray ball(s)
which provide complete coverage of the inside of vessels or discharged directly into product
transfer pipework via change piece.
2. The CIP flow velocity, temperature and required cleaning time must be verified and
documented for each cleaning validation test. Where CIP flow switches are installed verification
of correct alarm operation should be included.
2.3. Cleaning Agents
Cleaning Agents such as detergents and other chemical aids should only be used when the
cleaning process using water or solvents is not adequate. This may be due to insoluble materials,
equipment that is difficult to clean or environmental factors.
The selection of appropriate cleaning agents and their control should take into consideration
safety, product/equipment compatibility, vendor/material controls, and change control.
Only approved cleaning agents should be used which are of known composition, to permit
analytical measurement of residues and proven to be easily removable.
As there is usually no clinical data for detergent residues criteria of 10ppm is acceptable.
3. Cleaning Validation Procedure
3.1. Identify process, equipment and product type
Identify the process and the types of products being produced. Include these details in the
validation plan, validation protocol or change request, whichever is relevant for the work or
project being undertaken.
3.2. Check if Cleaning Validation is required
3.2.1. If the process or equipment is any of the following a cleaning validation assessment is
required:
· Holding vessel
· Mixing vessel
· Product transfer Line
· Other equipment, which performs an automated cleaning of product contact parts.
Title: Procedure For Cleaning Validation
3.2.2. New Processes/Equipment
Full validation testing is required in the majority of cases. This involves three tests of worst-case
product with the appropriate number of samples collected. Reduced testing is feasible if all of
the following conditions are met:
1. Identical process/equipment is installed.
2. Identical cleaning controls and procedures are achievable.
3. Worst-case product is the same as previously validated and acceptance criteria is the same
or higher than previously validated.
4. Cleaning validation data is available on the previously validated identical process.
An explanation for reduced testing should be described in the relevant Validation Plan, protocol
or change request for the process. Reduced testing may include reducing the number of tests and
reducing the sample locations to a few key critical areas known to be “critical sites” or “hot
spots”.
For changes to current cleaning processes and procedures the extent of retesting will in most
cases be reduced to a single test, this shall be stated and approved as part of the change request
process as in QMS SOP.
3.3. Select Worst-Case Product for Cleaning
For multi-product equipment it is not practical to validate cleaning of all products produced in a
particular process/equipment that has one cleaning process and where products are alike in
formulation and dosage form. In such cases, it is considered acceptable to select a worst-case
product to represent all products in the process for the purposes of cleaning validation. The
worst-case selection is based on a scientific justification i.e. the least soluble product produced
on a particular equipment or process. This product is then used to validate cleaning for that
process/equipment.
3.4. Select Product to use for acceptance criteria calculations
1. Calculation of acceptance criteria is to be based on the most toxic product within a group
of products produced in a given process.
2. Find the product within the product group, which has the lowest active toxicity value in
mg/kg, this is the most toxic product. This product will be used for the calculation of acceptance
criteria.
3.4.1. Example of selecting worst-case product and product for acceptance criteria calculations
Note - a low toxicity value means high toxicity, a high value means low toxicity.
Title: Procedure For Cleaning Validation
ActiveActive solubility in water (mg/mL)Active Toxicity (mg/kg)
16090
Active Active solubility in water Active Toxicity (mg/kg)
(mg/ml)
1 60 90
2 400 2800
3 70 0.30
From the above table of product data, Active 1 is the least soluble product in water since it has a
value of 60 mg/mL and is therefore the worst-case product in this group.
Active 3 is the most toxic product since it has the lowest toxicity value of 0.3 mg/kg. Therefore
the acceptance criteria is calculated using active 3 product data.
3.4.2. New Products Introduced to the Production Facility
If a new product is introduced into the facility the solubility and toxicity of the new product
should be compared against the current list of products in the same product type group and
following assessment should be made:
· If the new product is less soluble than the current least soluble product within the product
group then this product becomes the new worst-case product for that process. Cleaning
validation, including analytical method validation should be conducted for the new product.
· If the new product is more toxic than the most toxic product within the group then the
worst-case product does not change but the acceptance criteria to be applied to the worst-case
product must be recalculated according to the product data for the new product. If previous
validation data shows the results meet the new acceptance criteria no further cleaning validation
is required.
· If the solubility of the new product is not less and the toxicity not higher than the current
listed products then no cleaning validation is required for the new product.
3.5. Check if Analytical Method Validation is required
After selecting the worst-case product check if an analytical method is validated for that product.
If it is not, analytical method validation is required. Refer to Section 4 for procedure for
completing analytical method validation. If a method is validated proceed to step 3.6.
3.6. Calculate Acceptance Limits for Rinse and Swab Samples
Title: Procedure For Cleaning Validation
3.6.1. Basis of Calculations
The Lowest concentration of the most toxic active in a product group will be used in
combination with a safety factor of 1/1000th to calculate the acceptance limits as detailed below.
Once an agreed acceptance limit is established it is applied to an area/weight limit based on
factors such as batch size and calculated surface area of product contact. The final limits
calculated are then applied to the worst-case product in a product group.
3.6.2. Establishment of Acceptance Criteria
The most stringent of the following limits should be applied:
1. Absence of visible residues AND Maximum Allowable Carryover as calculated in section
3.6.3.
OR
2. Absence of visible residues AND No more than 10ppm carryover of the previous product will
appear in any subsequent batch.
Therefore option 2 is applied when the calculated Maximum Allowable Carryover (MAC) is
above 10ppm OR if there is no clinical data such as lowest concentration dose for the residue to
be calculated.
3.6.3. Calculating Maximum Allowable Carry Over (MAC)
When batch and single active concentration volumes for multi-product equipment
are known, the total MAC (in the entire processing system) is calculated as
follows:
MAC = (LC) x (SBS)
(SF) x (LVSD)
MAC = maximum allowable carry-over
LC = lowest concentration (in mg)
SBS = smallest batch size (in ml) made in the same equipment
LVSC = largest volume single concentration (active) of any product made in the same
equipment (in ml)
SF = safety factor = 1000. This is based on biological activity levels of 1/1000 of the
normal active concentration.
Example Calculation
Title: Procedure For Cleaning Validation
MAC = 0.25mg x 200,000mL = 25.0 mg
1000 x 2.0mL
Therefore the total quantity of residual product allowable in a subsequent production batch is
25.0 mg.
3.6.4. Calculating Acceptance Limits For Swabs
1. Take the calculated MAC for the product and divide this number by the total internal
surface area of the total product processing system, i.e. preparation and holding vessels +
pipework + filling machine. This figure is the amount of residue allowed throughout the entire
process, the assumption being that there is even distribution of product residue throughout the
process equipment.
2. Example calculation:
The allowed residue in the entire process = MAC/total surface area
= 25.0mg/56715 cm2
= 0.00044 mg/cm2 (MAC/cm2)
The overall surface area is derived from the following (only as example):
EQUIPMENT/S EQUIPMENT INTERNAL SURFACE AREA
(cm2)
Mixing tank 26745
Holding tank 26745
Transfer lines 965
Filling Machine 2260
Total 56715
This calculated value determines the amount of residual product allowed to remain on 1 square
centimetre of the equipment after cleaning. This value is then multiplied by the area to be
swabbed to give the allowed limit per swab sample.
If swabbing a 10 cm x10 cm (100 cm2) surface area and placing the swab in 25.0ml
of swabbing solution then the following applies:
Limit for swab sample = MAC/cm2 x Swab area
Volume of Swab Solution
Title: Procedure For Cleaning Validation
= 0.00044 mg/cm2 x 100 cm2 = 0.0088 mg/mL (8.8mg/ml or ppm per swab)
25.0 ml
In this case, swab sample results for 100 cm2 must be £ 8.8 mg/mL of active to prove that the
process is satisfactory. The value 0.00044 mg/cm2 was
derived from the MAC allowed per swab calculation above.
3.6.5. Calculating Acceptance Limits For Rinse Samples
1. Calculating required rinse volume:
Rinse recovery method validation tests have been conducted with a particular volume of solution
per surface area of the test material. This data is shown in the reference spreadsheet. From this
ratio of rinse volume to surface area apply this to the area of the process to be sampled. For
example, rinse recovery method validation records the ratio of rinse: surface area to be
20mL:100cm2, for a pipe of surface area 2500 cm2, the required volume is 20/100 X 2500 =
500mL (0.5L).
2. Calculating Limits
To calculate take the MAC and divide it by the total internal surface area of the processing
system. For example, using the MAC calculated on above, rinse sample limits are calculated as
follows:
Limit for rinse sample = (MAC/cm2) x Pipe or Vessel Surface Area *
Rinse Water Volume
= 0.00044 mg/cm2 x 965 cm2 = 0.00085 mg/mL or 0.85 ppm
500 mL
*actual surface area of the specific pipe or vessel being rinsed.
In this example a 500 mL rinse water sample of the transfer line, with a surface area of 965 cm2,
must be £ 0.85 ppm of residual active to prove the cleaning process is satisfactory.
3.6.6. LOD and LOQ Check
After calculating the final swab and rinse limits, check that the Limit of Quantitation (LOQ) for
the worst-case product is below the calculated acceptance criteria. If accurate analysis is
restricted by the limit of quantitation (LOQ), the limit of detection (LOD) can be used to provide
a pass/fail result.
Title: Procedure For Cleaning Validation
For example, if the LOQ is 700 parts per billion (ppb) and the LOD is 200 ppb, for a swab
sample with a failed result of 190 ppb, the laboratory would record a “FAIL”, as opposed to a
numeric value.
3.7. Flowchart – Cleaning Validation as annexure
4. Analytical Method Validation for Cleaning
4.1. Active Residue Analytical Testing
Complete protocol template “Cleaning, Active analysis method validation”.
The template is a test protocol that provides full details of the test objective, acceptance criteria
and the test method to be followed. This protocol should be used as a draft, any relevant changes
should be made prior to use and the final test protocol checked and authorised.
This test is conducted to determine the accuracy of measuring the active at concentrations above
and below the calculated acceptance criteria levels for cleaning. This is done simply by
preparing the relevant concentrations in volumetric flasks and analysing the samples by
Chromatographic procedures or by Total Organic Carbon. This tests the accuracy and precision
of the analytical method, without any effects of sampling recovery from surfaces by swabbing or
rinsing.
4.2. Rinsing Recovery Studies
Complete protocol template “Cleaning, Rinsing Method Validation.
The template is a test protocol that provides full details of the test objective, acceptance criteria
and the test method to be followed. This protocol should be used as a draft, any relevant changes
should be made prior to use and the final test protocol checked and authorised.
4.2.1. Purpose
Rinse recovery studies must be conducted for the specific product to be tested on the production
equipment. This must be completed before rinse samples can be taken from equipment surfaces.
This will ensure that the product can be recovered from the equipment surface with an adequate
recovery level.
Method validation studies shall determine the repeatability, reproducibility, and recovery of the
rinsing analysis from the equipment surfaces. If recovery results do not meet the acceptance
criteria a different solvent, or a larger rinsing volume may need to be used.
4.2.2. Rinsing Procedure
Rinse recovery studies are performed by spiking 316L stainless steel plates (or other production
material if more relevant for a given process). Product is evenly distributed onto the plate at
Title: Procedure For Cleaning Validation
concentrations above and below the acceptance criteria calculated in step 3.6, “Calculating limits
for Rinse and swab samples”.
The stainless steel plates must be large enough to allow a 10 cm x 10 cm surface area to be
rinsed. Allow the product to dry on the sample surface before rinsing. Rinse the plate with a
specified and accurate amount of water, i.e. by using a pipette. Collect the rinse water into a
beaker and analyse the rinsate as per analytical procedure using High Performance Liquid
Chromatography or by Total Organic Carbon.
TOC samples should always be analysed as soon as possible after sample collection. i.e. within
12 hours.
4.3. Swabbing Recovery Studies
Complete protocol template “Cleaning, Swabbing Method Validation”.
The template is a test protocol that provides full details of the test objective, acceptance criteria
and the test method to be followed. This protocol should be used as a draft, any relevant changes
should be made prior to use and the final test protocol checked and authorised.
4.3.1. Purpose
Swab recovery studies must be conducted for the specific product to be tested on the production
equipment. This must be completed before swab samples can be taken from equipment surfaces.
This will ensure that the product is adequately recovered from the equipment surface based on
the appropriate selection of swab material and solvent.
Method Validation studies shall determine the repeatability, reproducibility, and recovery of the
swabbing analysis from the equipment surfaces. If recovery results do not meet the acceptance
criteria, a different swab type, different solvent, or different swabbing method may need to be
used.
4.3.2. Swabbing Procedure
Swab recovery studies are performed by spiking 316L stainless steel plates (or other production
material if more relevant for a particular process). Product is evenly distributed onto the plate at
concentrations above and below the acceptance criteria calculated in step 3.6, “Calculating limits
for Rinse and swab samples”.
The stainless steel plates must be large enough to allow a 10cm x 10cm (100cm) surface area to
be swabbed. Allow the product to dry on the sample surface before swabbing.
Following is a recommended procedure to follow which has been shown to work well for
Method Validation tests:
Title: Procedure For Cleaning Validation
Place 2 swabs into 1 clean 40mL Total Organic Carbon (TOC) vial containing the required
volume of extraction solvent (usually 25mL). The 2 swabs are to be used for the same 10x10cm
surface and placed back into a single vial.
The swab surface is pressed on the side of the glassware to express excess water prior to use.
The swabbing is conducted covering the area in one direction then using the flip side of swab
surface swab in a perpendicular direction. Firmly press down on the swab handles to ensure
proper surface contact. The second swab is removed from the solvent and the first swab placed
back into the solvent.
The procedure is repeated with the second swab ensuring that the exact same 10cm x 10cm area
is swabbed; the swab is placed back into the solvent. The swab handles are cut with a clean pair
of scissors making certain that no foreign particles are introduced into the solution. The solution
is vortexed for 30 to 60 seconds.
TOC samples should be analysed as soon as possible after sample collection, i.e.
within 12 hours.
4.3.3. Why Two Swabs?
Two swabs are in most method validation cases necessary to give increased recovery from
equipment surfaces. The 1st swab may collect 70% of residue; the second clean swab is then
able to absorb the remainder more easily from the surface. Also when two swabs are then placed
inside the 40mL TOC sample vial this provides a greater mixing action than 1 swab when the
samples are vortexed.
4.4. Selection of Analytical Testing Instruments
For most applications Method Validation should be performed by using both TOC and HPLC
analysis. However if the product to be validated has no additional excipients containing organic
carbon other than the active ingredient then only HPLC needs to be used. If however it is
necessary to have the flexibility of using either TOC or HPLC analysis then both analytical tests
must be validated.
The detailed analytical test method used for quantifying the results must be recorded in the
analytical method validation files.
5. Cleaning Validation Test Protocols
To prepare the protocol use template “OQ Test Protocols”. The template provides an outline of a
test objective, acceptance criteria and the test method to be followed. This protocol should be
used as a draft version, any relevant changes for a particular project should be made prior to use
and the final test protocol checked and authorised by the relevant staff.
5.1. Test Objective
Title: Procedure For Cleaning Validation
Number of Tests Required
For new processes a minimum of three consecutive tests of the cleaning process must meet the
acceptance criteria for all swab and rinse samples in order to have a validated cleaning
process/procedure. Tests not meeting these criteria must be explained with a suitable
justification and corrective action i.e. Validation discrepancy.
5.2. Acceptance Criteria
Calculated swab and rinse limits for the product group as described in section 3.6.
5.3. Equipment Contamination
Whenever possible contamination of equipment should be under worst-case conditions i.e. leave
machine contaminated and drained overnight before cleaning. It is important that all product
contact areas are completely covered with the product contaminate.
5.4. Selection of Sample locations
Swab sampling locations should be selected for areas which are known to be difficult to clean
and/or theoretically maybe difficult to clean. The names of areas sampled must be clearly
indicated and defined in the results.
5.5. Sampling Procedures
Sampling procedures in most situations will involve rinse water and swab techniques. For
difficult to clean areas and when contact surfaces are physically accessible swabbing will be
used. Surfaces inaccessible to swab samples such as transfer pipes will be sampled using a pre-
determined volume of final rinse solution, usually water. Sampling procedures must be included
in individual Protocols. The procedure must indicate the sample materials required, the sample
solution (e.g. water type), the rinse volume or swab sample areas, and the sample locations.
5.5.1. Swab sampling
The swab method should be based on the procedure validated by the analytical laboratory. In
many cases the surface of production equipment will not be a flat stainless steel surface.
Therefore the swab must be done as close as practically possible to the validated swab procedure.
Non-standard swab areas
Where it is not possible to swab 100cm2 the actual area swabbed is recorded and an adjustment
to the acceptance limit is made. For example, if the swab area is only 50cm2 the limit is halved.
5.5.2. Rinse sampling
The rinse method should be based on the procedure validated by the Analytical laboratory.
Non-standard rinse volumes
Title: Procedure For Cleaning Validation
Where it is not possible to rinse to the required ratio of Rinse:Surface area, the actual volume
used is recorded and an adjustment to the acceptance limit is made. For example, if the rinse
volume calculated is 1L and 2L was required the limit is then halved.
5.6. Monitoring During Automated Cleaning Cycle
The main data required from any test is cleaning water flowrates, cleaning time and water
temperatures.
5.7. Collecting Rinse Samples
In some cases for each rinse sample both a chemical and microbiological sample is required, if
this is the case collect the microbiological sample 1st then aseptically transfer some of the
solution into a sample container for chemical testing.
For manual rinse samples the following precautions should be followed.
Containers for collecting samples i.e. sample jars, trays, buckets, etc. must be
clean and thoroughly rinsed with distilled water, especially when taking conductivity
measurements and for TOC analysis. For TOC testing use clean TOC vials or glass Schott
bottles. Any devices such as manual valves used to collect samples must be of a cleanable
design and always cleaned prior to use.
Containers used to pressure transfer water samples through product lines must also be clean and
rinsed thoroughly with Distilled water. For TOC testing it is important to collect a small sample
of the rinse water used as a blank sample to measure the background TOC.
5.8. Collecting Swab samples
The principles explained under rinse samples also apply to swab sampling. The swabbing
procedure must be based on the procedure validated as part of the analytical method validation.
The relevant file for method validation should be used as a basis for describing the swabbing
procedure in the test protocol.
5.9. Collecting Microbiological samples (bioburden)
The main requirements are that sample containers are pre-sterilised; the sample valves used are
clean and pre-sanitised by flushing with 80°C distilled water for 5 minutes.
5.10. Failed Results
Any failures of the rinse and swab samples must be dealt with by investigating the reasons for
the failure; making changes to procedures and then repeating the test. Sampling, testing, re-
sampling and re-testing the same equipment should not be conducted if test results continually
fail to meet the Acceptance Criteria. The “test until clean approach” or testing until the desired
results are obtained demonstrates the cleaning process is not in control. If this occurs an
Title: Procedure For Cleaning Validation
improvement to the cleaning procedure must be investigated. This may include alteration of
product contact materials such as flexible transfer tubing or improvement of the cleaning cycle,
extending the flushing time or removing potential dead-legs.
6. Change Control/Revalidation
6.1. Change Control
Before any changes to validated processes or process equipment are conducted the change
request form must be approved to determine whether it will impact on previous cleaning
validation studies (see SOP QMS-030.). If the change has the potential to decrease cleaning
effectiveness of product residuals, retesting will be required documenting the change and the
results from the new tests.
6.2. Revalidation
For automatic cleaning processes ongoing verification may not be required provided that the
process is validated such that it is proven to be in control and reproducible.
An assessment of any revalidation requirements will be made in the final Validation Report for a
given process.
