CLEANING VALIDATION

Arun Kumar Mathur

Pharma Consultant
Ph. 9833287686
BACKGROUND

• The cleaning of facilities and equipment is an important

measure to avoid contamination and cross
contamination.
• In compliance with the GMP regulations, cleaning is
performed and documented according to the described
procedures.
• In the past, cleaning effectiveness was often monitored
only visually. However, residuals of APIs and excipients
as well as of detergents are increasingly an issue in
inspections and audits.
• The success of cleaning procedures has to be validated.
In addition to the FDA “Guide to Inspection of Cleaning
Validation”, the PIC/S document PI 006 and Annex 15
address cleaning validation in a separate chapter.
Moreover, the ICH Guideline Q7 “GMP for APIs” also
requires cleaning validation.

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INTRODUCTION, PRINCIPLE AND OBJECTIVE

“Cleaning Validation is the methodology used to assure that a

cleaning process removes residues of the active pharmaceutical
ingredients of the product manufactured in a piece of equipment,
the cleaning aids utilized in the cleaning process and the
microbial attributes. All residues are removed to predetermined
levels to ensure the quality of the next product manufactured is
not compromised by waste from the previous product.”

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GENERAL REQUIREMENTS (FDA)

• FDA expects firms to have written procedures (SOP's) detailing the cleaning processes used
for various pieces of equipment. If firms have one cleaning process for cleaning between
different batches of the same product and use a different process for cleaning between
product changes, we expect the written procedures to address these different scenario.
Similarly, if firms have one process for removing water soluble residues and another process
for non-water soluble residues, the written procedure should address both scenarios and
make it clear when a given procedure is to be followed. Fluid bed dryer bags are another
example of equipment that is difficult to clean and is often dedicated to a specific product.
Any residues from the cleaning process itself (detergents, solvents, etc.) also have to be
removed from the equipment.
• FDA expects firms to have written general procedures on how cleaning processes will be
validated.
• FDA expects the general validation procedures to address who is responsible for performing
and approving the validation study, the acceptance criteria, and when revalidation will be
required.
• FDA expects firms to prepare specific written validation protocols in advance for the studies
to be performed on each manufacturing system or piece of equipment which should address
such issues as sampling procedures, and analytical methods to be used including the
sensitivity of those methods.
• FDA expects firms to conduct the validation studies in accordance with the protocols and to
document the results of studies.
• FDA expects a final validation report which is approved by management and which states
whether or not the cleaning process is valid. The data should support a conclusion that
residues have been reduced to an "acceptable level."
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 POSSIBLE CONTAMINANTS

• Pharmaceutical products can be

contaminated by a variety of
substances such as contaminants
associated with microbes, active
pharmaceutical ingredients (API) and
excipient residues of previous
products, residues of cleaning
agents, airborne materials such as
dust and particulate matter,
lubricants and ancillary material,
such as disinfectants, and
decomposition residues from:
• Product residue breakdown occasioned by. e.g. the use of
strong acids and alkalis during the cleaning process and
• Breakdown products of the detergents, acids and alkalis that
may be used as part of the cleaning process

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REGULATORY REQUIREMENTS FOR CLEANING VALIDATION:

21 CFR 211.67 that states: “Equipment and utensils be cleaned,

maintained and sanitized at appropriate intervals to prevent malfunctions
or contamination that would alter the safety, identity, strength, quality or
purity of the drug product”. According to this law each and every
pharmaceutical and food industry should follow the cleaning validation
program to avoid malfunctioning, contamination and cross contamination
of finished product.

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CLEANING VALIDATION PROGRAM:

Equipment cleaning validation may be

performed concurrently with actual production
steps during process development and
manufacturing. Validation program should be
continued through full scale commercial
production. The concept “Test-Until-Clean”
should be applied. This concept involves
cleaning, sampling and testing with repetition
of this sequence until an acceptable residue
limit is attained

A validation program generally encompasses at

least three consecutive successful replicate to
establish that the procedure is reproducibly
effective if the equipment of the similar size,
design and construction is cleaned by the same
procedure, studies need not be conducted on
each unit as long as a total of three successful
replicates are done on similar piece of
equipment; this concept is known as
equipment grouping.
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SAMPLING PROCEURE:

There are two methods of sampling.

1. Direct surface sampling (swab
method):
This method of sampling is the most
commonly used and involves taking
an inert material (e.g. cotton wool)
on the end of a probe (referred to as
“swab”) and rubbing it methodically
across a surface. The swabs are
added to the dilution solvent and
these solvents are analyzed by
suitable analytical instruments for
the presence of residue of previous
products per given area. i.e. 60-100
square inch

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SWAB SAMPLING
The location from which the sample is taken should take into consideration the
composition of the equipment (e.g. glass or steel) and the location (e.g. blades, tank
walls or fittings). Worst case location should be considered. Critical areas, i.e. those
hardest to clean, should be identified, particularly in large systems that employ semi
automatic or fully automatic clean-in-place systems.

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SWAB SAMPLING (MICRO)

For determination of the microbiological contamination on surfaces is to use sterile

cotton swabs moistened with sterile peptone water, water for injection, or phosphate
buffer. Using sterile forceps and aseptic technique, a predetermined area is wiped
with a sterile swab. The swab is then aseptically transferred to a sterile tube
containing a suitable diluent. The tube is then agitated to suspend any viable
microorganisms and aliquots are placed in a semisolid medium to obtain quantitative
results.

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RINSE SAMPLING

2. Rinse samples (Indirect method):

In this method, a measured area of clean surface is rinsed or washed with solvent
and the solvent is collected and tested for traces of contaminants. This method
allows sampling of a large surface, of areas that are inaccessible or that cannot be
routinely disassembled and provides an overall picture. It is also suitable for
checking the residue of cleaning agents, e.g. detergents.
Rinse sampling method should be used in combination with other sampling methods
such as surface sampling. There should be evidence that samples are accurately
recovered. For example a recovery of > 80% is considered good, > 50% reasonable
and < 50% questionable

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RINSE SAMPLING

Rinse Sampling involves passing a known volume of solvent over a

large area and analyzing the recovery solution.
• The solvent rinse occurs after cleaning has been completed.
• This method is not as direct as swabbing but will cover the
entire surface area (and parts inaccessible to swabs).
• It is important to ensure chosen solvent has appropriate
recovery for residues being quantified.
• This method allows much greater ease of sampling than
swabbing.
• A reduced no of samples are required to generate a carryover
figure.

• Sampling and testing of rinse samples for residual active ingredient is a commonly adopted method
to evaluate cleanliness. This is a fairly convenient method in many cases and requires control over
the solvent used for rinsing, the contact time and mixing involved. The solvent used should be
selected based on the solubility of the active ingredients and should either simulate a subsequent
batch of product or at least provide adequate solubility.

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DETERGENTS

The efficiency of cleaning procedures for the removal of detergent residues should be
evaluated. Acceptable limits should be defined for levels of detergent after cleaning.
Ideally, there should be no residues detected.
The possibility of detergent breakdown should be considered when validating
cleaning procedures. Detergents that have persistent residues such as cationic
detergents which adhere very strongly to glass and are difficult to remove, should be
avoided where possible.

The composition of detergents should be known to the manufacturer. If such

information is not available, alternative detergents should be selected whose
composition can be defined. The manufacturer should ensure that he is notified by
the detergent supplier of any critical changes in the formulation of the detergent

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ANALYTICAL METHODS
Determine the specificity and sensitivity of the analytical method used to detect
residuals or contaminants. With advances in analytical technology, residues from the
manufacturing and cleaning processes can be detected at very low levels. If levels of
contamination or residual are not detected, it does not mean that there is no residual
contaminant present after cleaning. It only means that levels of contaminant greater
than the sensitivity or detection limit of the analytical method are not present in the
sample. The firm should challenge the analytical method in combination with the
sampling method(s) used to show that contaminants can be recovered from the
equipment surface and at what level, i.e. 50% recovery, 90%, etc. This is necessary
before any conclusions can be made based on the sample results. A negative test
may also be the result of poor sampling technique.

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ANALYTICAL METHODS

The analytical methods should be validated before the cleaning validation is performed
and the methods chosen should detect residuals or contaminants specific for the
substances being assayed at an appropriate level of cleanliness (sensitivity). The
detection limit for each analytical method should be sufficiently sensitive to detect the
established acceptable level of the residue or contaminants.
Some of the analytical methods which can be used for the analysis of cleaning validation
samples include:

− HPLC
− GC
− HPTLC
− TOC
− UV spectroscopy
− pH
− Conductivity
− ELISA
These methods can be used alone or in combination depending upon the analysis
required.

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RECOVERY

Calculation of Recovery factor: (1 to 1.25)

% recovery shall not be less than 80% unless otherwise specified and justified in
individual
protocol of analytical method validation. Recovery factor shall be calculated as follows:
100
Recovery factor = ---------------
% Recovery

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ESTABLISHMENT OF LIMITS:

The rationale for selecting limits for product residues should be logically based on a
consideration of the materials involved and their therapeutic dose. The limit should
be practical, achievable and verifiable.
The approach for setting limits can be:

• Product specific cleaning validation for all products

• Grouping into product families and choosing a “worst case” product
• Grouping products according to risk, e.g. very soluble products, products with similar
potency, highly toxic or difficult to detect products.

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ACCEPTANCE CRITERIA:

S.N Testing Acceptance criteria

o. Parameter
1 Physical The equipment should be visually clean. i.e. no residue should be
determination visible on equipment after cleaning.
2 Chemical a) NMT 0.1% of the normal therapeutic dose of any product to
determination appear in the maximum daily dose of the subsequent product.
b) NMT 10 ppm of any product to appear in the next product
(basis for heavy metals in starting materials).
c) For certain allergic ingredients, penicillin, cephalosporin or
potent steroids and cyto-toxins, the limit should be below the limit
of detection by best available analytical methods
3 Microbial Total aerobic counts
contamination a) NMT 10 cfu/100 ml by rinse method.
b) NMT 5 cfu/25 cm
2
by swab method.

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NOEL AND MACO CALCULATIONS

NOEL = No Observed Effect Level

MACO = Maximum Allowable Carry Over

NOEL is determined to calculate MACO in Cleaning Validation. NOEL is the amount of

drug in mg that does not have any effect on human health. It is calculated by using
Lethal Dose 50 of the drug. LD50 is the amount of drug that kills the 50% population
of the animals used in the test.

NOEL = (LD50 X 70kg) / 2000

where,
LD 50 - Lethal Dose
70kg - Average adult weight
2000 - Constant
For Example, if any drug has LD50 331mg/kg then NOEL will be
NOEL = 331x70/2000
11.58 mg

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NOEL AND MACO CALCULATIONS

This value of NOEL (11.58mg) will be used to calculate MACO

MACO = (NOEL x MBS) / (SF x TDD)

Where,
MBS - Maximum Batch Size
SF - Safety Factor (1000 for oral drugs)
TDD - Total Daily Dose of next product
For Example, if TDD of next product is 500mg and batch size is 400kg, then MACO
will be calculated as:
MACO = (11.58 x 400000000 mg) / (1000 x 500 mg)
= 4632000000/500000
= 9264 mg
= 9.264 g

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MACO

MACO Calculation (Option 2)

by considering 0.1% safety factor

Daily therapeutic dose of product A (in mg) Minimum batch size of product B (in
mg)
Limit (in mg) = ----------------------------------------------------------X ----------------------------------------------------------
1000 Max daily therapeutic dose of product B (in
mg)

Where,
Product A = Product manufactured before cleaning
Product B = Next product after cleaning

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ACCEPTANCE CRITERIA FOR SWAB SAMPLES

For considering 10 ppm as acceptance criteria

The quantity equivalent to 10 mg/L of the batch size is considered as the acceptance
criteria of 10 ppm.
Calculation of acceptance criteria for Swab samples

Limit (PPM) = MACO x 1000/C x D/V

Where,
C = Cumulative surface area of the equipment used (in [Link])

V = Volume of solvent used to dispense swab

D = Swabbed surface area in [Link]
1000 is the multiplication factor to convert value in mcg from mg

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ACCEPTANCE CRITERIA FOR RINSE SOLUTION

Calculation of acceptance criteria for Rinse samples

1000 1
Limit (PPM) = MACO x ----------- x ------
C V

Where,
C = Cumulative surface area of the equipment used (in sq. cm)
V = Volume of solvent used for rinse of the same in mL/ [Link] of equipment
1000 is the multiplication factor to convert value in mcg from mg

For example for MACO value of 9264mg an area of 2500 sq./cm is washed with 350 ml of solvent

Limit (ppm) = 9264 x 1000/2500 x 1/350 = 10.58 ppm

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REVALIDATION

Revalidation of cleaning procedure is required if any of the following occur:

• Cleaning procedure is changed
• Raw materials are changed
• Change in formulation
• New detergent
• Change in analytical method for determination of residue
• Major non-traceable contamination occurrence
• Failure during cleaning verification/validation

Revalidation of cleaning procedure shall be performed on a minimum of three cleaning cycles.

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BIBLIOGRAPHY

REFERENCES:
1) The validation of cleaning processes, Pharmaceutical Technology – S.W. Harder
2) Cleaning Validation: An overall perspective – Jenkins M, Vanderweilen AJ
3) ICH: Good Manufacturing Practice guideline for Active Pharmaceutical Ingredients.
4) Pharmaceutical Quality Assurance: Cleaning Validation – Manohar A. Potdar
5) Determining cleaning validation limits for pharmaceutical manufacturing operations,
Pharmaceutical Technology – Fourman, G.L. and Mullen, M.V.
6) Points to consider in the validation of equipment cleaning procedures, Journal of Parenteral
science and Technology - James Agalloco
7) Process Analytical Technology and Cleaning – Verghese, G. and Lopolito, P.
8) Cleaning Technology for Pharmaceutical Manufacturing – LeBlanc, Destin A. et al.
9) Cleaning Validation in the Pharmaceutical Industry by Mowafak Nassani,Ph.D.

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 Thanks !

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