ISSN 0973 – 6921 ; E – ISSN 2319 – 5983 J. Environ. Res. Develop.

Journal of Environmental Research And Development Vol. 13 No. 03, January-March 2019
Review Paper (NS-I)
CLEANING ANALYTICAL METHOD VALIDATION
Pushpa Agrawal* and Raghu M.G.
Department of Biotechnology, C.M.R.T.U, R.V.College of Engineering Campus, Bangalore,
Karnataka (INDIA)

Received August 14, 2018 Accepted January 03, 2019

ABSTRACT
In pharmaceutical industry, the main purpose of cleaning validation is to enhance the efficiency
and consistency of cleaning process in pharmaceutical production equipment or to avoid cross
contamination due to incorrect cleaning of equipment, apparatus, processing area or the starting
material, this will result in severe risks for the consumers. So it is required to validate the
cleaning procedures to ensure safety, quality, efficacy of the next batches of drug product and
regulatory requirements in finished product manufacturing units. By proper cleaning of
equipment, apparatus as well as the processing area contamination can be minimized.
Pharmaceutical manufacturing companies need to validate their cleaning process to make sure
compliance with cGMP regulations. The collective data used to conclude the success of a
cleaning validation is built upon both the effective assessment of the manufacturing plant and the
robustness of the validated analytical method. In order to ensure the safety of the consumer,
there must be a high degree of assurance in the results obtained by analytical methods in order to
confirm the absence of residues at the predefined limits on the different equipment surfaces. This
review focused on the different types of parameters performed in cleaning analytical method
validation adapted by pharmaceutical industry.
Key Words : Cleaning validation, Analytical method, Swab, Rinse, Cleaning limit

INTRODUCTION In current situation Food and Drug

Administration (FDA) inspections shows
Cleaning means to make any article, piece of
equipment and area free from marks, dirt, or scrutiny and big interest in cleaning validation
any unwanted substances.1,2 Pharmaceutical in the pharmaceutical industry. The major
product will be contaminated by different requirement of current good manufacturing
materials such as residue of previously used practice (cGMP) during various steps of a
active pharmaceutical ingredient (API), manufacturing process is the adoption of
cleaning agents, excipients and dust validation of procedures used to clean the
particles3-5. The inappropriate cleaning of equipment7,8. During federal agency audits,
equipment’s can lead to contamination and auditors were expecting to see a functioning
cross contamination of pharmaceutical cleaning validation program with proper
products. So, there is a requirement of properly records in place during their inspections. The
cleaning of equipment apparatus and cleanliness of the equipment before using is
processing area in the manufacturing not a new concept, it’s a requirement of
pharmaceutical industries. Cleaning validation cGMP9. The equally important requirement
is a documented process that demonstrates the that it also be hygienic is many times confused
efficiency and consistency in cleaning a by the word, clean. In response to the question
pharmaceutical production equipment’s. Cross “what is clean,” the FDA published a guidance
contamination must be avoided within the document: the 2004 FDA “Guide to
pharmaceutical industry at all times and
Inspections Validation of Cleaning Processes.”
successful cleaning validation ensures that
The FDA’s guide to inspections, cleaning
patients safety due to cross contamination6.
validation helps in analytical investigation of a
*Author for correspondence cleaning procedure.
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ISSN 0973 – 6921 ; E – ISSN 2319 – 5983 J. Environ. Res. Develop.
Journal of Environmental Research And Development Vol. 13 No. 03, January-March 2019
The purpose of cleaning validation is to verify approved with acceptance residual limits. This
the cleaning methods for removal of residues of determines that the laboratory equipment and
previous product, cleaning agents, detergents, methods are capable of evaluating with
preservatives, or/and microbial contaminants. validation parameters, according to approved
Cleaning validation fulfils the necessities of validated analytical methods to quantify the
regulatory agencies and keeps product quality small amount of residual contaminant.
and safety of end user10. The success of cleaning By mimicking the same active product and the
procedures is evaluated by collecting samples same material surface of production equipment
from cleaned product contact surfaces using would be appropriate for the validation of the
suitable sampling methods11-13 (e.g., rinsing, analytical method. Preparation of different blank
swabbing). Before their use, it is expected that solutions, serial dilutions of active standard
these sampling procedures should be qualified to preparations could be used along with a placebo
confirm that they are acceptable for the meant preparation containing the additives and excipi-
purpose and don't result in false results14. The ents preparations would be suitable for mimic-
overall data obtained from validated analytical king the quantification of active residual during
methods is used as a supportive document for the validation of the analytical method. Before
successful completion of cleaning validation. It’s collection of samples from the cleaned equip-
essential that these obtained results are really ment, it is essential to verify the efficiency of the
representative as patient safety is based upon the swab materials used in the analytical method.
absent of residues from equipment’s15-17. Development of test method
With the manufacturing process of pharmace-
OBJECTIVE
utical product, the analytical method plays major
The objective of the cleaning validation is to significance when measuring the cleanliness of
validate the efficiency of the cleaning pharmaceutical manufacturing equipment. In
procedure for removal of drug residues, order to obtain the satisfactory results, the
preservatives, degradation products, cleaning developed analytical method should be
agents or excipients, as well as the control of consistent, accurate and robust. The analytical
potential microbial contaminants18,19. method developed should be specific to the
It is required to validate cleaning procedures analyte, and also able to quantify at the
for the following reasons: prescribed residue limits.
● Pharmaceutical products and drug The residual limits are set by the individual
substances can be contaminated by other pharmaceutical industries as per there prescribed
finished products, cleaning agent and standard operating procedure. The regulatory
microbial contamination.
authorities do not set specification limits for
● It is regulatory requirement in
specific products. While setting the residual limit
manufacturing of pharmaceutical product
with respect to a given pharmaceutical product,
that concern is the same-assurance that
few important things has to be taken care i.e.,
equipment is clean from contamination
residual limits should be logical, they should be
and that product quality and safety are
practical and achievable and verifiable
retained.
● To reuse the equipment Cleaning analytical method validation
● It is also compliance point of view and A validation protocol is necessary to define the
assure quality from an internal audits. specific items and activities that will constitute a
● To protect product reliability and integrity cleaning validation study. This procedure
describes the validation of analytical methods
DISCUSSION used in cleaning validation studies of
Analytical method manufacturing and packaging equipment. The
The analytical method should be validated for the validation of an analytical method is required to
determination of residual amount of active establish the performance characteristics of the
substance20. Prior starting of the cleaning method as it is intended to be used. Performance
validation, a proper analytical cleaning method characteristics are expressed in terms of
protocol should be prepared, reviewed and analytical parameters.
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Journal of Environmental Research And Development Vol. 13 No. 03, January-March 2019
Procedure Dry the stainless steel container. Prepare a stock
Analytical method : A complete description solution in solvent or solvent mixture. Disperse
of a rinse and swab preparation, test procedure, the appropriate volume of the stock solution
sufficiently detailed to enable qualified person to evenly on the inner surface of the stainless steel
replicate it. It includes all important operational vessel. Minimum of two different concentration
parameters and specific instructions for the levels, in triplicate are required. The
preparation of reagents, performance of system concentration levels should cover approximately
suitability test, precaution, and explicit formulas
lower to upper limit of the cleaning limit. Dry the
for calculation and reporting of test results.
vessel, after each time spiking, in air and the
Analytical performance parameters :
Analytical parameters defined by the current rinse with appropriate volume of diluent.
United States Pharmacopeia (USP) which Analyze the solutions according to the method.
identifies the performance characteristics of the Method precision (Swab recovery)
method based on its designated category. These Pre-wash the swabs specified in the method.
include, but are not limited to, instrument Prepare a stock solution in solvent or solvent
precision, linearity, accuracy (recovery) from mixture. Disperse the appropriate volume of the
rinse and swab, accuracy (recovery) from stock solution evenly on the head of the swab.
respective surface coupons, method range, Minimum of two different concentration levels,
method precision, quantitation limit, specificity in triplicate and 100% of cleaning limit for six
(if applicable), solution stability and surface times are required. The concentration levels
stability. should cover approximately lower to upper limit
Extraction solvent or solvent mixture : The of the cleaning limit. Add the specified volume
solvent or solvent mixture used to extract the of extraction solvent into each test tube and
residual drug substance from the swabs.
proceed as directed in the method. Analyze the
Product contact surfaces : Equipment surfaces
solutions according to the method.
that product(s) may contact during
manufacturing and packaging. Stainless steel Calculate the recovery of analyte using the
(SS) is a major contact surface. Product contact following equation:
surfaces other than stainless steel are dependent Amount of analyte
on manufacturing equipment train. recovered
% Recover : X 100
Cleaning limit : A calculated amount of residue Amount added
product A which can remain on the equipment Intermediate precision
and in turn, contaminate a maximum daily dose A second analyst repeats the recovery of the
of product B, fallows product A on production analyte from the spiked swabs for six times at
train. 100% cleaning limit and analyze the solutions
Validation Parameters using different HPLC system, a different column
Specificity on a different day.
Follow the individual methods to prepare Linearity
rinse/swab blank solution and rinse/swab blank Prepare linearity solutions at minimum five
solution after rinsing/swabbing surfaces using the different concentrations that cover the QL to
same number and type of swabs. Prepare a
upper limit of cleaning limit. When possible, the
placebo in extraction solvent at an appropriate
highest concentration should be based on the
concentration. Prepare the detergent solution at
an appropriate concentration. Each of the analyte maximum allowable carry over (MACO)
to be cleaned from the production/packaging calculations for drug residue and 10 µg/mL
equipment using detergent should pass the (default upper limit) for cleaning agent. Inject
specificity test updated detergent cleaning each of the linearity solution in duplicate into the
verification protocol. HPLC system and record the results. Plot the
Rinse recovery regression line of peak area versus analyte
Pre-wash the stainless steel container with water, concentration. Calculate the regression
acetonitrile and respective diluents for 3 times. coefficient and bias at 100% level.

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Journal of Environmental Research And Development Vol. 13 No. 03, January-March 2019
Detection limit (DL) and Quantitation limit If same HPLC system is used, calculate the
(QL) percent difference for standard and test
Approximately dilute the linearity solution solution as follows:
near to the QL and DL level concentration to Time zero peak area –
determine signal to noise ratio (s/n). Perform Stability time point
% Difference :
six replicate injections of the QL solution. peak area X 100
Surface accuracy Time zero peak area
A stainless steel plate with a rough surface is If different HPLC system is used, calculate the
used for the recovery study. Prepare a stock percent difference for test solution as follows
solution in solvent or solvent mixture. Disperse Time zero result –
the appropriate volume of the stock solution Stability time point
% Difference :
evenly onto the area of the stainless steel plate result X 100
specified in the method. Allow the plates to air Time zero result
dry and visually examine the plates for the Swab equivalency
evidence of residual material and document the Swab equivalency should be established by
results. Swab the area using the number of performing accuracy study at 100% target
swabs specifies in the method. Transfer the level on stainless steel surface in triplicate
swabs in to respective test tube. Add the using different swabs than specified in the
specified volume of extraction solvent into method.
each test tube and proceed as directed in the Recovery from other surfaces
method. Analyze the solutions according to the Validation of product contact surface other
method. Minimum of three different than stainless steel surface depends on the
concentration levels, in triplicate are required equipment train used in manufacturing.
for all types of surfaces for accuracy and CONCLUSION
ruggedness. The concentration levels should
The production equipment’s used in
cover from the lowest concentration of the
pharmaceutical industry should be free from
accuracy study from the stainless steel plate
individual contamination or cross
(QL) to approximately upper level of the
contamination and pharmaceutical products
cleaning limit.
should be safe for the consumer/end user. At
Calculate the recovery of analyte using the
following equation: the level of 100%, it is practically not possible
Amount of analyte to prove that manufacturing equipment is
% Recovery : recovered “clean”. However, it is likely to prove that the
X 100 traces of left over residue, over the equipment
Amount added
Solution stability parts, are within a predefined acceptable limit
This study is to demonstrate the stability of the and an analytical method is capable of
standard and test solutions stored at the room detecting and quantifying these trace levels.
temperature. Cleaning validation proves that the
Note: A recovery solution from a spiked swab contamination levels have been reduced below
and/or stainless steel plate can be used as the residual acceptance limits. Cleaning validation
test solution for this study. Prepare a standard activity covers active residue identification,
solution and test solution according to the residue detection, sampling method selection,
method. Store the working standard solution the establishment of residue acceptance
and test solution at a room temperature in a criteria, analytical method validation, recovery
closed container on bench top. Analyze the studies, and the identification of equipment
standard solution and test solution according to parts in direct contact with the pharmaceutical
the method at an initial time point. Analyze the finished product. With adopting these things in
same solution (minimum of 3 time points) at cleaning validation we can minimize any kind
intervals up to a maximum of prescribed days. of contamination or cross contamination.
Calculate the stability of the solution for each The final analytical technique is able to
time point investigated. completely cover the requirements of the
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Journal of Environmental Research And Development Vol. 13 No. 03, January-March 2019
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(2016).
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