Cleaning and Disinfection Validation

John Holah and David Childs, Holchem Laboratories Ltd, Lancashire, United Kingdom
© 2019 Elsevier Inc. All rights reserved.

Introduction 1
Validation 1
Monitoring 1
Verification 2
Validation Drivers 2
Which Cleaning Objectives Should Be Validated 2
Concurrent, Prospective or Retrospective Studies 3
Validation Prerequisites 4
Validation Team 4
Validation Scope 5
Qualification of the Food Processing Equipment to Be Cleaned 6
Qualification of the Cleaning Programme 7
Soiling Characteristics 10
Environment, Equipment and Product Sampling Techniques 10
Analytical Tests 11
Health and Safety, Legislative and Performance Requirements 13
Validation Process 13
Validation Protocol 13
Validation 14
Validation Interpretation 14
Monitoring, Verification and Cleaning Programme Target Setting 16
Review 16
References 17

Introduction

Food and drink should be safe to the consumer by being free from physical, chemical and biological contamination. Various
hazards can contaminate processed food, such as microorganisms and their toxins, the previous product (including allergens),
cleaning residues and machinery lubricants. To control such hazards and protect the food from contamination, the manufacturing
equipment and surrounding environment should be routinely and effectively cleaned. Cleaning and disinfection is thus crucial to
maintain and guarantee food safety and comply with legislative and due diligence systems requirements.
As cleaning and disinfection is so important in ensuring food safety, good manufacturing practice would dictate that there
should be evidence of the fundamental ability of the cleaning and disinfection programme to control identified food safety hazards,
and additional evidence or records that the programme is working or has worked on a day-to-day basis. Several terms have been
used (and misused) to describe this evidence and the following terms are proposed.

Validation
Documented evidence of capability – the designed and documented cleaning and disinfection programme is fit for the purpose of
meeting the programme’s objectives, primarily concerned with hazard control, when performed as documented and in all perceived
operational conditions.

Monitoring
Real-time assessment of effectiveness – at appropriate times during the cleaning and disinfection programme, observations and
measurements assess the performance of the programme in a time-frame that allows the next stages of the programme to commence
(e.g. whether equipment surfaces have been cleaned and rinsed sufficiently for disinfectants to be applied) or corrective actions to be
undertaken.

Reference Module in Food Sciences https://doi.org/10.1016/B978-0-08-100596-5.21207-9 1

2 Cleaning and Disinfection Validation

Verification
Determination of compliance – evidence that the cleaning and disinfection programme undertaken at a particular date and time,
had met all of its objectives, including the control of any identified hazards. Evidence for verification is not time critical and typically
comprises laboratory based tests for e.g. specific pathogens, allergens or DNA, that may require several days before a result is gener-
ated and communicated.
Thus, once a cleaning programme has been validated, it is then routinely applied, monitored and verified.

Validation Drivers
The evidence that cleaning and disinfection programmes, further shortened to cleaning programmes, can control identified food
safety hazards is thus provided by validation, but why has this term and requirement recently come to attention within the food
industry?
Historically, cleaning validation started in the pharmaceutical industry in the late 1980s/early 1990s, when the Food and Drug
Administration in the USA began to view cleaning as a ‘process’, and as such it needed to be validated similarly to process validation
(Walsh, 2011). The driver for pharmaceutical cleaning validation was to ensure that there was no cross-contamination of e.g. peni-
cillins, steroids or hormones between previous and post-cleaning pharmaceutical batches, and a number of pharmaceutical valida-
tion guidelines has been published (Active Pharmaceutical Ingredients Committee (APIC), 2014; Food and Drug Administration
(FDA), 2016).
As a Hazard Analysis Critical Control Point (HACCP) prerequisite (Alimentarius, 2007), cleaning and disinfection has always
been regarded as important and, as part of good manufacturing practice, its effectiveness has traditionally been actively assessed and
reviewed. Indeed, audit schemes based on the Global Food Safety initiative (GFSI) have traditionally recognised the concept of vali-
dation and current standards (at the time of writing), such as the British Retail Consortium (BRC) Global Standard for Food Safety
Issue 8, provide guidance on the need for validation. Clause 4.11.3 states that: Where cleaning procedures are part of a defined prereq-
uisite plan to control the risk of a specific hazard, the cleaning and disinfection procedures and their frequency shall be validated and records
maintained.
Cleaning and disinfection is a key part of a pathogen control programme and is responsible for freeing the process environment
from pathogens that had previously entered the manufacturing area via raw materials or breakdowns in high hygiene area barrier
controls e.g. entry changing procedures or packaging decontamination. It is now recognised that for many pathogens, e.g. Listeria in
ready-to-eat (RTE) products, failures in product decontamination processes controlled by CCPs are rare. Failures in prerequisite
controls prior to the decontamination stage, which create a high pathogen challenge to the decontamination process (e.g. prior
to the washing of fruit and fresh produce), or failures post-decontamination, causing cross-contamination to the decontaminated
product, e.g. cooked sliced meats, are the major causes of pathogen contamination of RTE products. The importance of prerequisites
in the control of pathogens has been recognised (Holah et al., 2011) and failures in cleaning programmes are often blamed for
pathogen contamination of RTE products. Validation of microbial control via cleaning and disinfection has thus received attention,
particularly as the cases of Listeria and other pathogens in the human population are increasing in Europe (European Food Safety
Authority (EFSA), 2016).
Perhaps the major focus on cleaning validation has come about because of the rise of importance of allergens in the food
industry (Arrowsmith and Brown, 2009). In factories in which allergen containing and allergen free products are manufactured,
cleaning is the major control to ensure that allergens are removed from the processing line and environment prior to the production
of allergen-free products. Also, analytically and in practical factory-based validation trials, allergens are measurable: there is an
obvious presence of allergens in the food soils prior to cleaning (they are a product ingredient) and thus their removal after cleaning
can be measured and demonstrated. This is not the case with pathogens, however, as the likelihood of practically detecting a path-
ogen on surfaces or in food products pre and post cleaning in a validation trial would be extremely rare.
Finally, there can also be a legal requirement for the control of some cleaning and disinfection chemicals in foodstuffs. For
example, legislation in Europe has established a series of maximum residue limits (MRLs) in foodstuffs for pesticides (EC 396/
2005), and two previously used pesticides, didecyldimethylammonium chloride (DDAC) and benzalkonium chloride (BAC),
which are primarily used in quaternary ammonium compound-based disinfectants, have had specific MRLs applied (EU No
1119/2014). If these disinfectants formed part of a cleaning programme, their presence in subsequent food products processed
on the line that had been cleaned and disinfected, at levels above their described MRLs, would not be legally acceptable.

Which Cleaning Objectives Should Be Validated

The objectives of a cleaning programme have been previously described (Holah, 2018) as follows, to:
(1) provide a safe working environment for employees;
(2) provide a clean working environment for employees;
(3) extend the life of, and prevent damage to, equipment and services;
(4) maintain plant operating parameters during food processing including heat transfer and flow parameters;
(5) remove materials that could lead to foreign body contamination or could provide food or shelter for pests;
(6) remove food soils that would be detrimental to the organoleptic quality of subsequent production runs.
 Cleaning and Disinfection Validation 3

(7) remove DNA to prevent e.g. one meat species (e.g. pork) contamination of subsequent production runs (e.g. beef) in a factory
processing multiple animal species;
(8) remove spoilage microorganisms to maintain or potentially extend product shelf-life;
(9) remove allergens to prevent contamination of subsequent production runs;
(10) remove pathogens that may contaminate food products.
Aligned to the above objectives, the need for validation is summarised in Table 1, which has three key outputs.
1. Where the cleaning programme is designed to control a hazard, it must always be validated.
2. Where the cleaning programme is designed to control a brand protection issue, validation is at the discretion of the food
manufacturer.
3. Where the cleaning programme is designed to control a quality or process control objective, validation is not generally advo-
cated. It could also be argued that where the end-point of the cleaning objective can be visualised, or the removal of food soil can
be rapidly assessed (e.g. by ATP or protein tests), validation is the equivalent to monitoring for these cleaning programmes.

Concurrent, Prospective or Retrospective Studies

Concurrent validation is a planned event, on existing process lines and products, to generate experimental data that can be recorded
as evidence of the satisfactory performance of the cleaning programme. It is undertaken with the overriding principle that validation
should cover all known or anticipated worst case scenarios, such as:

• the most difficult part of the equipment to clean

• the most difficult to remove food soil
• the worst-case production schedule resulting in a final soil that is difficult to remove
• the lowest acceptable range within the prescribed conditions of the cleaning programme, e.g. water temperature, chemical
strength, contact times
• the shortest cleaning window
• the longest wait between production finishing and cleaning commencing

Table 1 Validation, monitoring and verification requirements

Cleaning objective Validation/Monitoring/Verification requirements

9–10 Hazard Control Validation essential to meet food safety best practice and
• Microbiological pathogens (Listeria, E. coli, Salmonella etc.) GFSI requirements.
• Allergens Monitoring/verification essential
• Chemical residues (not a cleaning objective but a hazard arising
from poor cleaning)
7–8 Brand Protection Validation is not essential for food safety but could be
• Microbiological spoilage (Pseudomonas, C. estertheticum etc.) customer driven for brand protection. There may also be
• DNA (e.g. pork in a beef product) legislative implicationsa
• Vegetarian (e.g. meat in a product labelled ‘Suitable for Validation is subject to analytical techniques being
vegetarians’ available to assess all the specific brand protection
• Organic (e.g. non-organic components in a product labelled as analytes.
‘Organic’) Monitoring/verification essential
• GMO (e.g. GMO components in a product labelled as ‘Free from
GMO’s)
• Religious (Halal, Kosher) (e.g. non-Halal components in a product
labelled as ‘Halal’)
1–6 Quality & Process Control Unlikely to require validation, particularly for open
• Quality (prevention of carry-over of previous product into equipment where the endpoint of visual cleanliness is
subsequent product) easily observed.
• Organoleptic (strong taints or colours) Could be considered for closed process equipment where
• Foreign body and pest prevention a visual endpoint may not be observed.
• Process performance (length of time before loss of process Monitoring/Verification to be determined as required
performance e.g. heat exchangers, length of time before cleaning
is necessary)
• Process safety (e.g. prevention of fire)
• Equipment life extension
• General cleanliness for safe and effective staff performance
a
Local legislation may require that there is maximum percentage of a previous product (e.g. horse meat) in a subsequent product (e.g. beef) during mixed species processing on the
same production lines. For example, the Food Standards Agency set a threshold of 1% horse or pork in subsequent food products as an indicator of good manufacturing practices in
mixed species processing, which includes cleaning and disinfection (FSA, 2014) .
4 Cleaning and Disinfection Validation

Table 2 The pros and cons of prospective and retrospective validation

Validation type Retrospective Concurrent

Pros • Data already exists • Predictive of all potential operational circumstances as

• Minimal additional cost to analyse such data based on worst case scenarios
Cons • Historically, worst case scenarios may not have been • Additional costs
encountered or sampled • Limited data
• Time to completion

If validation is undertaken under the known and anticipated worst case scenarios, then theoretically, all subsequent, routine clean-
ing scenarios should equally be validated.
Prospective validation is similar to concurrent validation, except that it is undertaken on newly developed process lines, e.g.
following the construction of a factory or a refurbished area, as part of a commissioning process and, ideally, before food products
from these lines are placed on the market. The same principle that validation should cover all known or anticipated worst case
scenarios is adopted, however some information may not be well established, such as:

• the worst-case production schedule resulting in a final soil that is difficult to remove;
• the shortest cleaning window;
• the longest wait between production finishing and cleaning commencing.
Because of the likely unknown production variables, and thus the difficulty in determining worst-case scenarios, prospective vali-
dation may have to be initially reviewed more frequently than concurrent validation.
Retrospective validation is an evaluation of a cleaning programme’s performance historically, based on all available monitoring
and verification records. Consideration is given to the consistency of the cleaning programme in meeting its hazard objectives, often
described as key performance indicators (KPIs), and the variability of monitoring and verification results.
Prospective validation has to be undertaken as part of the commissioning of new builds/refurbishment projects. The pros and
cons of concurrent and retrospective validation are outlined in Table 2.
Concurrent validation, undertaken correctly, is the preferred option as, if carried out in defined worst case scenarios, it should be
representative of all likely production, soiling and cleaning programme variations. It is, however, an additional cost exercise and,
even if repeated a number of times, produces only a fraction of the dataset of historical verification records.
Retrospective validation may not always be reflective of worst case scenarios, particularly as routine verification sampling points
may not be the hardest areas to clean (which may be more difficult to access routinely). However, the robustness of the historical
data increases with the length of the time that the cleaning programme has been in operation, the knowledge around the perfor-
mance of the cleaning programme and how post cleaning verification samples were taken. If the food process and the cleaning pro-
gramme have been little changed over several years, it may be argued that ‘worst cases scenarios’ may well have occurred and been
sampled. This is much less likely to have happened in a cleaning programme that has been running for a few months where, for
example, the effect of any seasonal variations would be unknown. The quality of historical results increases with evidence, e.g. if
the hardest places to clean were consistently sampled; records exist of pre-cleaning conditions, e.g. length of production time
and products processed, and records of cleaning parameters, e.g. detergent concentration, temperature and cleaning times are avail-
able. Equally, when records identified poor cleaning programme results, the cause of such poor performance had been established.
If all such records are available, retrospective validation can be a powerful tool.

Validation Prerequisites

The European Hygienic Engineering Design Group (EHEDG), 2016 advocates a series of necessary prerequisites which have to be
undertaken prior to the validation process.
These activities include the need to ‘qualify’ the validation team, the process line or cleaning object to be validated and the clean-
ing programme. In addition, the worst-case soiling conditions need to be identified, sampling sites and sampling methods deter-
mined, and analytical tests confirmed.
Any legality issues surrounding the validation process (e.g. for the disinfectants used) and health and safety aspects in under-
taking the work, should also be considered.

Validation Team
Validation of cleaning programmes is the responsibility of the food manufacturer. Guidance can be sought from equipment manu-
facturers, chemical suppliers, cleaning contractors and other external consultants with the relevant experience and knowledge, where
required. Therefore, creating the right validation team is key and the approach is very similar to developing a HACCP team, with
members all having the correct levels of experience and qualifications. Indeed, and as cleaning and disinfection is only one of
 Cleaning and Disinfection Validation 5

a number of HACCP prerequisites that controls hazards, food manufacturers may have appointed a generic validation team as part
of their food safety programme.
A cleaning programme validation team should comprise of:

• Senior management support, who ultimately will sign-off the validation as complete and enforce it
• A Team Leader who will coordinate all necessary pre-meetings and practical validation exercises
• Representatives from all areas of the factory with ownership of the cleaning objectives, the cleaning window, the undertaking of
the cleaning programme and the maintenance of cleaning parameters (equipment, water temperatures, dosing equipment etc.).
Such representatives could include:
• Production
• Engineering
• Technical
• Hygiene
• Hazard specialist (microbiologist, chemist)
• External support from other stakeholders who have an interest in the execution of a successful validation programme
• Equipment manufacturers
• Cleaning chemical suppliers
• Contract cleaners (if used)
• Internal or external analytical testing laboratory
• Customers
• Consultant
• A scribe to take notes and write up the Validation Report.
The validation team should initially be trained in the concepts of validation and on-going monitoring and verification, and be
familiar with the design and outputs of the validation process.
The validation of cleaning programmes is not technically difficult and does not require specialist equipment. As such, it is invari-
ably undertaken in-house and is not contracted out to specialist third party consultants, e.g. as for thermal process validation.

Validation Scope
The first step of the validation process is to focus on the scope of the cleaning validation programme and the objectives it is seeking
to validate.
The scope may be simplistic, such as a cleaning programme performed on a process line to control a pathogen, or relatively
complex, such as the cleaning and disinfection of the process line, utensils, product containers and transport systems and operatives
personal protective equipment (PPE) associated with the changeover between an allergenic product run followed by an allergen free
product run.
As noted in Table 2, the primary objectives of the cleaning programme being validated will be the control of hazards, with
secondary objectives being the control of brand protection issues. There may be the need to validate cleaning programmes control-
ling quality and process control attributes where visual cleanliness cannot simply be verified. This is particularly the case in CIP
cleans where the most difficult to clean areas of the process equipment may not be easily accessed to inspect and where the potential
degree of soiling in these areas could adversely affect quality or processing objectives.
The validation scope should, therefore, consider the following:

• Define the product(s) and process line(s) that the validation will cover.
• Define the cleaning programme type
• Interim
• End-of-production
• Periodic
• Define the process/equipment/utensils to be cleaned
• Open surfaces
• CIP circuits
• Product contact surfaces
• Non-product contact surfaces
• Environmental surfaces
• Automated cleaning systems – e.g. tray wash, tote bin wash
• Others – e.g. on-site laundry
• Define the cleaning programme’s objectives
• Visual cleanliness
• Absence of pathogens
• Freedom of allergens below the lower detection limit of the allergen detection kit used
• Freedom of a chemical residue below any imposed maximum residue limit
6 Cleaning and Disinfection Validation

• Freedom of DNA below any legislative limits

• Desired TVC, ATP or protein levels determined from historical practices (if such information is available)
• Define the cleaning programmes hazards
• What surfaces are impacted by the product soil to be cleaned – process line, associated equipment e.g. weigh scales,
ingredient containers, product probes or sampling equipment, operative’s PPE?
• Where could hazard residues of the product being cleaned be transferred to e.g. adjacent production lines?
The majority of the cleaning programme’s objectives can be easily assessed, but for more specific objectives, such as a particular
allergen or disinfectant residue, it should be first checked if analytical tests are available to quantity these analytes before the vali-
dation scope is accepted.

Qualification of the Food Processing Equipment to Be Cleaned

The process of equipment validation has two prime purposes. Firstly, to assess whether the process equipment that the
cleaning programme is intended for is fundamentally cleanable. In other words, has it been appropriately hygienically designed
and is it fit for purpose. Secondly, to identify the features or geometries of the equipment that are likely the most difficult to
clean.
Hygienic design of equipment is a prerequisite for equipment fundamental cleanability. All food processing machinery sold into
the European Community is required by law to be hygienically designed and basic guidance as to what such hygienic design criteria
needs to be met is given by the Machinery Directive (2006/42/EC). The requirements of 2006/42/EC are met if the machine and
associated components comply with EN 1672 (2009)
For countries where hygienic design is not legislated, guidance is available from:

• Any national hygienic design standards

• The international hygienic design standard, ISO 14159 (2008)
• Guideline documents from the European Hygienic Engineering Design Group (EHEDG) www.ehedg.org
• Guideline documents from the 3-A Sanitary Standards Inc. www.3-a.org
• International trade associations such as the International Dairy Federation. https://www.fil-idf.org/publications/
• National trade associations such as the USA based North American Meat Association https://www.meatinstitute.org/ and
Grocery Manufacturers Association https://www.gmaonline.org/
Following assessment, if an item of equipment has identified poor areas of hygienic design, these may not effectively be cleanable,
with any cleaning programme undertaken. Poor areas of hygienic design can be mitigated by:

• Replacing the machine with one of a more hygienic design (an expensive option!)
• Refurbishing the machine to remove or modify the poor design
• Creating better access to the poorly designed area for cleaning chemicals. This may involve e.g. providing access platforms,
removing guards, further dismantling of the equipment
Hygienic design principles primarily cover the design of the equipment and its components as it is delivered to the customer by the
equipment manufacturer. Any modifications of the equipment by the food manufacturer, e.g. in linking the machine to a process
line or the replacement of parts and fluids during maintenance (seals, gaskets, lubricants and transfer fluids), becomes the respon-
sibility of the food manufacturer. Replacement parts and fluids should be food grade and be traceable.
The determination of the areas of the equipment that are most difficult to clean presents a worst-case scenario. It is assumed that
if these areas of the equipment are cleaned to the objectives of the validation, all other surfaces of the equipment will be similarly
cleaned.
Areas that may be difficult to clean can be assessed via examining plans of the equipment and by visual inspection. Ideally for
visual inspection, the equipment should be out of production, pre-cleaned and engineers should be available to undertake any
dismantling necessary or aid in access.
Identified areas that are likely to be the most difficult to clean then become the area(s) that will be assessed and sampled for
cleanability during the validation process. Records/photographs should be undertaken of these areas to be included in the valida-
tion protocol (see later in text).
If the areas deemed likely to be the hardest to clean are not capable of being accessed, or are unsafe to access, a compromise may
have to be made between the place(s) that is likely hardest to clean with places that are hard to clean but are practicably accessible.
At the same time as identifying the areas that are most likely to be difficult to clean, it is also worthwhile establishing the areas
that are to be used for future monitoring and verification purposes, which may not be the same. As part of the validation protocol, it
may be possible to determine the relationship between areas thought to be the hardest to clean but practically/routinely inacces-
sible, with those areas chosen for monitoring and verification purposes.
This will not be possible for pathogen or allergen analysis, as both these areas should be free from pathogens/below the limit of
allergen detection, but may be possible for quantifiable assessment techniques such as rapid Adenosine Triphosphate (ATP)
detection.
 Cleaning and Disinfection Validation 7

Qualification of the Cleaning Programme

Cleaning programmes are not universal and should be chosen and developed to meet the required cleaning objectives for the soil
and the equipment to be cleaned.
The parameters that need to be determined, for both open and closed equipment cleaning include:

• the water quality and pressure

• the soil characteristics of the products processed
• any chemical approval and legality requirements
• any chemical/material incompatibilities
• the basic cleaning parameters – manual, chemical, temperature, automation
For open surface cleaning, once these parameters have been determined, trials can begin to develop a best practice cleaning pro-
gramme. Such trials may manipulate chemical types, solution concentrations and temperatures and chemical application methods
to ostensibly meet the cleaning objectives, prior to validation. The cleaning programme should be characterised so that it is defined
and repeatable. This could include:

• Length of cleaning window

• Number of operatives
• Chemical concentrations
• Chemical solution temperatures
• Rinsing water pressures and temperatures
• Cleaning procedures – gross solids removal/rinse/detergent application/rinse/disinfectant application/rinse (if required)
• Chemical safety information
• Required personal protective equipment (PPE)
A typical way of expressing this information is a cleaning instruction card (CIC), or a sanitation standard operating procedure
(SSOP) a basic example of which is shown in Fig. 1. At his stage the CIC is in draft form and provides sufficient detail for the clean-
ing programme to be repeatable. Basic training of the cleaning operatives against this CIC can then be undertaken and recorded.
Prior to validation it is also essential to ensure that all cleaning equipment is serviced, calibrated and working correctly, including
dosing pumps, spray cleaning equipment and water boilers.
For the cleaning of closed plants, validation is more complicated as two parameters have to be first established. Firstly, the ability
of the CIP system to fundamentally be able to consistently clean all parts of the plant must be established. Secondly, as for open
surface cleaning, a suitable cleaning programme must be developed and demonstrated to work for the specific soil types to be
encountered.
The basic parameters of the CIP system that need to be established could include:

• Flowrate through pipework is
1.5 m/s

• Flowrate through vessels is  10 m3/h
• For vessels fitted with spray balls, ensure the scavenge pumps are set to prevent vessel pooling
• For vessels fitted with rotary spray jets, define the spray time necessary to completely cover all internal surfaces of the vessel
• Ensure there is no shadowing inside vessels from spray patterns
• All appropriate instrumentation is in place
• CIP solutions are filtered
• The CIP circuit is drainable
• It is possible to CIP key aspects of the CIP system, e.g. final rinse tanks
• If appropriate, process equipment surfaces can be raised to pasteurisation or sterilisation temperatures
• Separate CIP systems should be in place for pre and post pasteurised product flows
• CIP and product flows are absolutely separated. If separation between product and CIP flows is via valves, there are always two
valves between product and CIP flows, with a visual flow to drain between the valves
After it has been established that the CIP set is designed to the recommended best practice design and is likely to achieve the desired
cleaning target, the development of the cleaning programme can be undertaken, together with its operational parameters.
Cleaning and disinfection parameters include:

• Cleaning window
• Solution temperatures
• Chemical types and concentrations
• Circulation times of all CIP clean/rinse/disinfection cycles are recorded
• CIP programme (rinse, caustic clean, rinse, acid clean, rinse, disinfection, rinse).
• CIP programmes are documented
8 Cleaning and Disinfection Validation

With respect to operational parameters, it is necessary to determine the relationship between the SCADA displays and environ-
mental reality including that:

• Flow and line diagrams are correct

• All valves and probes/instruments etc. are identified and calibrated
• What is described by the CIP software is occurring on the CIP and process lines e.g. valves defined by the software as open are
actually open on the plant
• All parts of the CIP system are cleaned, e.g. lifting valve seats

Figure 1 Example of a cleaning instruction card (CIC).

 Cleaning and Disinfection Validation 9

Figure 1 (continued).

As for open plant cleaning, a draft CIP programme should be documented after it has been trialled and established to meet its clean-
ing objectives. Basic training of the CIP operatives against this programme can then be undertaken and recorded. It is also essential
to ensure that all CIP equipment is serviced, calibrated and working correctly, including circulation pumps, spray cleaning devices,
probes, instrumentation and heating devices.
For open plant cleaning, acceptable cleaning parameters may be stated as a range in the CIC documentation, e.g. detergent
should be dosed in the range 3%–5% v/v, rinse temperature should be 45–50  C. Validation should be undertaken at the worst
cases scenario, in this case with the detergent dosed at 3% and the rinse temperature at 45  C. This is not usually the case for
CIP validation as the cleaning parameters are usually more accurately defined and the CIP programme may not operate if these
defined parameters are not met – e.g. from feedback from temperature or conductivity probes.
10 Cleaning and Disinfection Validation

Soiling Characteristics
The purpose of this step is to determine the soiling worst-case scenario. Two aspects of soiling have to be considered; the difficulty of
the soil to be removed and the level of hazard in the soil. Soil adherence characteristics can be influenced by:

• Strongest adhering soil

• Longest production run time
• Highest process temperature
• Product scheduling – is there a scenario where several stock keeping units (sku’s) of the product likely to be the strongest
adhering are running consecutively?
• Time before cleaning – is there a soil that remains on equipment time for long periods before it is cleaned – e.g. over the weekend
The level of the hazard in the product applies to pathogens and allergens primarily. For example, in a sandwich factory, sandwiches
containing fully cooked ingredients are less likely to contain pathogens than those which contain ingredients that have been
washed, e.g. fresh produce ingredients. Therefore, validation for freedom of pathogens should be undertaken on products with
the highest pathogen risk. For allergens, it may simply be which product sku has the highest level of allergen as a product ingredient?
Again, product scheduling during production may have an influence, particularly if several sku’s containing the same allergen are
run consecutively with no interim cleaning between sku’s.
The choice of target soil will be site dependent. One site may have a product soil that is particularly difficult to remove, whilst
another site may have a product with a high allergen ingredient content and may be interested in the validation of many associated
cleaning events related to the production of that product (e.g. food contact surfaces of the process line, automated tray washing of
ingredient containers delivered to the line, operatives’ PPE laundering).
Determination of the worst-case soiling conditions has two purposes. Firstly, and most obviously, to validate that the cleaning
programme can meet its objectives against the most difficult to remove soiling. Secondly, to cut down on the number of validations
required. For example, if a food manufacturer has a number of allergens that are controlled by the same cleaning programme, vali-
dating the programme for the worst-case scenario for a single allergen (highest allergen presence, most difficult to clean soil), theo-
retically confers validation for all allergens used.

Environment, Equipment and Product Sampling Techniques

The type of sampling undertaken to assess whether the objective of the cleaning programme has been met will depend on the hazard
or analyte being assessed. The vast majority of the sampling techniques used are well established and universal in the food industry
and are routinely used for monitoring and verification purposes.
There may be the need to use some specialist sampling techniques or instruments such as borescopes (Fig. 2) which can be used
to view poorly accessible parts of open equipment or inside vessels or pipework of closed processing equipment.
It is likely that the primary objective of all cleaning validations is visual cleanliness. Indeed, it can be argued that if visual clean-
liness is not established, it is pointless doing any further surface or product testing. Techniques for visual cleanliness include:

• Visual cleanliness described as absence of soil particles visible to the naked eye
• Absence of odours which may be reflective of fresh or aged soiling
• Absence of surface films. These can be detected as stains if a clean cloth is wiped across the surface, or the way that rinse water
droplets are retained or otherwise on the surface

Figure 2 Examples of borescopes allowing a range of distances that they can penetrate into equipment or drainage systems to capture an image of
the internal surfaces.
 Cleaning and Disinfection Validation 11

• Use of visible light or UV light-based torches to assess equipment with poor natural lighting
• Use of angled mirrors for assessing difficult to reach areas (e.g. underneath equipment) or borescopes for assessing pipework and
vessels
Visual cleanliness assessment is usually directed to assessing whether all residues of natural food soils have been removed. It is
possible to artificially soil equipment with soils that e.g. fluoresce under UV light to establish whether equipment has been cleaned;
residues are then detected post cleaning via UV light torches. However, these techniques are more readily used to either determine
which areas of equipment are more difficult to clean (in establishing worst-case scenarios) or for the training of cleaning operatives
(to ensure all areas of the equipment are cleaned), rather than in validation studies.
If visual cleanliness has been achieved, the techniques that can be used to assess specific hazards or general hygiene indicators
include.

• Direct surface sampling

• Swabs – targeted towards difficult to clean, unhygienic features of small dimensions – for allergens, microorganisms and
DNA
• Wipes/sponges – targeted to maximise hazard detection over large surface areas – primarily for pathogens
• Contact plates/dipsticks – targeted to small, flat surface areas. Dipsticks may also be used to sample rinses
• Specific swabs – supplied by test kit manufactures for the detection of e.g. ATP or protein detection for the use with their
specific detection systems or instruments
• Lateral flow strips (LFD) – primarily for allergens
• Indirect surface sampling
• Final rinses – CIP systems for allergens, microorganisms, chemical residues, DNA
• Flushes (undertaken in the dry food industries using small volumes of cheap ingredient raw materials, often abrasive such as
salt) – used between product streams - for allergens, microorganisms, chemical residues, DNA
• Air sampling
• Open Petri dishes – collection devices such as Petri dishes can be placed close to where aerosolised hazards may be an issue,
e.g. adjacent process lines. Aerosols landing in the Petri dish from the cleaning activity can then be assessed directly or
indirectly from the dish surface for the desired hazard
Wherever possible, direct sampling of surfaces should be used for validation purposes. Indirect sampling via rinses or flushes may
give rise to an ease in collecting a sample but have inherent sampling problems. Flushes and rinses:

• May not be able to remove soils adhering in e.g. cracks and crevices within the surface that is flushed.
• The flush or rinse may only be able to remove some of the soil components and not necessarily all of those components. The
flush or rinse is thus a measure of what it can remove, not necessarily the cleanliness of the surface.
• Soils may not be evenly distributed in the rinse or flush – soils may be flushed off the surfaces in an ad-hoc pattern
• There is a natural dilution of hazards in the soil removed by the large volume of rinse or flush agent. This may dilute the hazard
below the limit of detection of the analytical technique used for its quantification.
For enclosed surfaces that are difficult to access directly (e.g. vessels and pipe systems), it is more accurate to dismantle the
equipment at the point likely to be hardest to clean, or temporarily fit appropriate sampling devices at these points, for the
validation process. If this is not possible, flushes and rinses may have to be used for validation purposes. Whenever it is
possible, direct sampling should be undertaken along with indirect flushes and rinses during the validation process, so that
a correlation between them can be established, and then flushes or rinses can be used subsequently for monitoring or verifi-
cation purposes.
For some validation exercises, e.g. with an objective to control chemical residues, allergens or DNA, sampling of the subsequent
product to be manufactured will be necessary. It is suggested that for open systems, the first product down the line will flush out any
hazards left after the cleaning programme and thus the first product down the line only should be tested. For closed systems,
however, experience with allergens has shown that there may be absorption into the first, middle and final product down the
line. This is because any soil residues left in the production equipment are likely to be in dead zones, where cleaning fluid or product
flow is minimal (Fig. 3). Residues from soils in these areas of poor hygienic design are likely to be slowly released throughout the
production process.

Analytical Tests
The prime purpose of the validation exercise is to establish that the cleaning programme can control the specific hazard or brand
protection issue that is the objective of the programme. In general, analytical tests to determine whether the hazard objective of the
cleaning programme has been met, in product and on environmental surfaces, should be specific, sensitive, representative, and
reproducible.

• Microbiological tests for pathogens should be pathogen specific. Indicator organisms e.g. the use of Enterobacteriaceae as an
indicator for Salmonella, may not be appropriate
12 Cleaning and Disinfection Validation

Figure 3 Product soil remaining in a poor hygienic design, dead zone, where fluid flow is minimal. Arrows indicate flow direction of product and
cleaning fluids. Soil will interact with subsequent product throughout the process run, or until the soil is removed.

Figure 4 A qualitive lateral flow device suitable for use in food processing environments, in which a liquid sample (from a swab) is added to the
sample area (left hand side) and flows to the right, to give an indication of the presence or absence of the allergen.

• Allergen tests can be qualitative or quantitative

• Quantitative laboratory based enzyme linked immunosorbent assay (ELISA) tests should be used wherever possible for
product validation
• Qualitative lateral flow devices (LFD) are preferred for environmental surface validation (Fig. 4) though laboratory based
ELISA tests can also be used
• DNA tests should be qualitative via PCR
• Advice on tests for cleaning/disinfectant residues on surfaces or final rinses should be sought from the cleaning chemical supplier
The secondary purpose of the validation exercise is to correlate the results from the specific hazard detection tests (which tend to be
expensive) to the more general analytical techniques that are going to be used for monitoring and verification purposes (which tend
to be cheaper). The validation exercise can also be used to set target levels for these monitoring and verification techniques.
In general, analytical tests to determine the performance of the cleaning programme via general hygiene indicators on environ-
mental surfaces should be easy to use and interpret, cost effective, sensitive, representative, and reproducible.

• Microbiological tests as a general hygiene indicator should ideally assess all microorganisms possible, e.g. a total viable count
(TVC) or a yeast and mould count
• More specific organisms can be chosen if they also may have an impact on the product, e.g. Pseudomonas spp. counts as these may
be indicative of potential spoilage issues post cleaning.
• Other organism groups such as Enterobacteriaceae are more usefully used in environmental sampling during production (rather
than post cleaning) to indicate changes in the food processing environment which may lead to the proliferation of pathogens
• Allergen lateral flow devices (LFD) are routinely used for monitoring and verifying cleaning performance for specific allergens
• Soil residues containing adenosine triphosphate (ATP) - Rapid hygiene tests detecting ATP are universal and are an excellent
quantitative way of detecting residues of most food product soils following cleaning (Fig. 5).
• Soil residues containing protein - Rapid hygiene tests detecting proteins are also universal and are an excellent qualitative way of
detecting residues of food product soils containing protein following cleaning.
• Soil residues containing protein - More sensitive protein detection techniques may have a correlation to the presence of allergens
or meat species post cleaning
• Soil residues containing meat species - A relatively small number of tests kits are available that can detect defined meat species by
e.g. ELISA
All analytical tests should have appropriate controls. For example:

• For allergen testing, positive controls should be established to ensure that the target allergen in the allergen containing food
product can be detected in that food product matrix.
 Cleaning and Disinfection Validation 13

Figure 5 An ATP luminometer which provides a rapid assessment of the amount of ATP present in the organic material removed from the sampled
surface by the swab, after the reagents within the swab have been activated to create luminescence.

• To a lesser extent this may also apply to the detection of the target allergen on food contact surfaces that the food product matrix
has contacted.
• For microbiological testing, controls should be undertaken for
• Neutralisation of any disinfectant residues present
• Demonstration that the transport media and transport/storage times will maintain the viability of any pathogens sampled
• The effect of any cleaning/disinfectant residues present in the sample matrix on the sensitivity of the analytical detection
technique should be established (for allergen, DNA and ATP tests). Note, wherever possible, analysis should be undertaken after
the detergent rinse and before disinfection, for all hazards for which the disinfection action is not relevant as a control.
• Selected contract laboratories should have the desired level of accreditation and undertake appropriate proficiency tests.

Health and Safety, Legislative and Performance Requirements

Prior to validation, all health and safety and legislative aspects of the cleaning programme should be undertaken including:-

• Consider the need for risk assessments for any dismantling or access activities of the equipment, to sample areas identified as the
most difficult to clean
• Have all risk assessments been completed for operatives to safely undertake the cleaning programme
• Do all chemicals meet required Health and Safety, chemical and food legislation? For example in Europe, are the disinfectants
formulated from (approved) active biocides listed in ECHA Article 95 (2018), which lists relevant substances and suppliers in
accordance with Article 95 of the Biocidal Products Regulation 334/2014
• Are all chemicals compatible with the plant materials of construction?
• Do chemical disinfectants meet all performance requirements with respect to disinfectant efficacy (e.g. approval to European
standard disinfectant test methods EN1276 or EN 13697), toxicity and tainting

Validation Process
Validation Protocol
The validation protocol describes the scope of the validation, the cleaning programme validated, the validation process, the results
of all analytical tests recording the cleaning programme undertaken (temperatures, chemical concentrations, circulation times etc.),
the results of all analytical tests recording the cleaning programme’s performance, the analysis of the validation and, if successful,
the sign-off to indicate that the validation was successful.
14 Cleaning and Disinfection Validation

The validation protocol should consider:

• Scope
• Cleaning and disinfection objectives
• Validation team and responsibilities
• Worst case scenarios
• Difficult to clean areas
• Minimum acceptable cleaning programme parameters
• Most difficult to remove product soil/soil with the highest level of hazard
• Risk assessments related to undertaking the cleaning and sampling
• Defined cleaning and disinfection procedure
• Sampling points and analytical methods
• Cleaning programme parameters achieved
• Sign-off sheets/photos/records
• Cleaning measurements (times, temperatures, concentrations, flow etc.)
• Number of cleaning cycles to be performed
• Corrective actions if objectives not met
• Records of analyses and interpretation
• Objective target setting
• Monitoring and verification methods to be adopted
• Sign-off statement
• Date of next review
Ideally a validation record template should be developed which can be used as an aide memoire, a record sheet for all recorded
information and tests results, and a signed-off record of the validation status of the clean/process line etc.

Validation
The number of occasions on which the validation exercise should be repeated should be established by risk assessment, but
a minimum of three occasions is recommended, according to the protocol as shown in Fig. 6:
Consideration should be given to the timings of the validation runs, which might include:

• The condition of the process line equipment and how quickly it becomes worn to an extent that affects cleanability
• Seasonal influences on e.g. food product ingredient quality, seasonal products or external temperatures
• Production influences with respect to production lengths, process conditions and cleaning windows
• Product scheduling influences
Following the satisfactory completion of the validation, the sign-off statement can be completed in the validation record template.

Validation Interpretation
The interpretation of the validation results for the hazard of concern may be relatively straightforward or quite complex, dependent
on the analyte measured.
For pathogen control, an absence of pathogens post cleaning is the only acceptable result. If a pathogen is detected post clean,
a revue of the cleaning programme must be undertaken.
However, given that the likelihood of a pathogen being present prior to cleaning is very low, the chance of a pathogen detection
due to the failure of a cleaning programme is extremely low. General microbiological counts are thus often used to give an indica-
tion of the success of the cleaning programme. Cleaning and disinfection together is a unit process which typical reduces the micro-
bial population on a surface by 5–6 log orders (Holah, 2014). Therefore, if the starting levels of microorganisms prior to cleaning
is <106 cfu, cleaning and disinfection should achieve a post clean count of <10 cfu per unit. If the starting levels are 107 cfu, clean-
ing and disinfection should achieve a count of 10–100 cfu etc.
Freedom of allergen in the subsequent batch down the line as measured by ELISA is a fundamental requirement of successful
validation. If an allergen is present in the subsequent batch, the cleaning method should be modified until freedom of allergen is
achieved. If this is not possible, the use of ‘may contain’ labelling should be considered.
Freedom of allergen on surfaces following cleaning by ELISA would be seen as the best possible scenario and provides high confi-
dence in the ability of the process line to be freed from allergens from the previous batch. Freedom of allergen on surfaces following
cleaning by LFD would be seen as an acceptable scenario and provides confidence in the ability of the process line to be freed from
allergens from the previous batch. It also provides confidence in the use of LFDs for subsequent monitoring and verification of
allergen cleaning procedures.
Detection of allergen on surfaces following cleaning by ELISA would require a risk assessment to be undertaken to ascertain if the
detectable allergen present on the surface is likely to be a significant risk to the subsequent batch. Detection of allergen on surfaces
 Cleaning and Disinfection Validation 15

Figure 6 Validation process schematic.

following cleaning by LFD requires the cleaning method to be modified until freedom of allergen is achieved (by LFD). If this is not
possible, and following a risk assessment, the use of ‘may contain’ labelling should be considered.
Meat species detection is generally undertaken to verify good manufacturing practices (i.e. <1% transfer of one species from the
previous batch to the subsequent batch). If batches are of 1000 kg, there needs to be <10 kg left from the proceeding product to
ensure <1% transfer. Cleaning to visual cleanliness between batches is probably sufficient to achieve this, though if assurance
beyond visual cleanliness is required, DNA testing is capable of some quantification in verifying residual DNA present.
To uphold ‘free-from’ claims for brand protection, however, is more difficult. Polymerase chain reaction (PCR) DNA tests have
a suggested limit of detection of 1–2 pg (109g). Is this sufficient? If no DNA is detected in a food product by this method, then there
is high confidence that any target DNA in the sample must be infinitesimally small. But if 10 pg was detected, is this of practical
concern?
Care must be taken when using such DNA techniques, however. To use the lower limit of detection (1–2 pg) as an objective of
a cleaning programme when cleaning between different meat species batches is inappropriate. No cleaning programme is currently
designed, or expected, to give a >9 log order removal of a soil from a surface.
16 Cleaning and Disinfection Validation

Monitoring, Verification and Cleaning Programme Target Setting

Following a successful validation, cleaning programmes should be routinely monitored and verified. Some of the monitoring and
verification targets will be compulsory, e.g. absence of pathogens post clean.
However, the generated data can be used to set target levels of general hygiene indicators e.g. ATP Relative Light Unit (RLU)
counts or microbiological Total Viable Counts (TVC) counts. Target levels for subsequent cleaning programmes could be set as
the average results of the validation plus any ‘comfort factor’. For example, if an average final rinse water ATP reading was 100
RLU, which was obtained under strictly applied and monitored validation cleaning conditions, a target ATP RLU value could be
set as 125 RLU for routine CIP operations. If many data points are available, for example in retrospective validation, a more statis-
tical approach could be adopted, e.g. a target set at three times the standard deviation of the data set, if the data was assumed to be
normally distributed.
For the verification of cleaning programmes designed to control allergens, it may be possible to routinely verify that the objective
of the clean had been met via ATP or protein measurements, rather than allergen LFDs. However, for this to be acceptable, it must be
demonstrated during the validation process that there is still detectable ATP or protein present on the surfaces when the validation
has established the absence of detectable allergen residues (via LFD or laboratory based ELISA). If this is possible, ATP or protein
could be used e.g. daily whilst allergen LFDs could be used e.g. monthly.
With respect to the (inaccessible) areas that are hardest to clean, as noted during equipment qualification, it may be possible to
determine the relationship between such areas and those areas chosen for monitoring and verification purposes. For example, vali-
dation protocol results may record ATP values of 100 RLU in the most difficult to clean areas and 50 RLU in the areas to be routinely
accessed for verification purposes. Post validation, if a target of <50 RLU is set for routine verification, then the non-accessible, most
difficult to clean areas of the equipment will likely have received an acceptable clean.
Ongoing monitoring of the cleaning programme can be undertaken by, for example:

• Visual cleanliness sign-off sheets

• Measurement of the critical cleaning parameters established in the validation, e.g.
• Chemical concentrations via titration/conductivity probes.
• Cleaning/circulation times
• Circulation temperatures – defined on the CIP return. Cleaning times commence on the achievement of the desired
temperature at the point of CIP return.
• Flow rates
• ATP of swabs or rinses. Readings can be taken prior to disinfection (has the plant been cleaned sufficiently for disinfection to
effectively take place) or after disinfection (the total cleanliness of the plant following cleaning and disinfection)
• LFDs for defined allergens
• Rapid protein analysis as a potential measure of allergen absence
Ongoing verification of the cleaning programme can be undertaken by, for example:

• Microbiological sampling of surfaces and rinse waters.

• Microbiological sampling of the first product through the line following the CIP programme.
• Allergen lateral flow strips
• Air sampling
• Auditing
• Visual cleanliness
• Cleaning and disinfection methods
• Hygiene Management System including Cleaning Instruction Cards
• Operative training records
• Trending and reviewing verification records

Review
The validity of the validation should be reviewed at least annually. Other triggers for review may include:

• Following any change in:

• Food ingredients
• Food products
• Process parameters
• Equipment
• Cleaning
• Disinfection
• Historical trends of the key monitoring and verification parameters (table or curves) indicating a reduction in cleaning efficiency
• An increase in deviations (alarms) that have occurred during the CIP cycle
 Cleaning and Disinfection Validation 17

• Any changes in the consumption of water and steam (the CIP station must be provided with flow meters), detergent and
disinfectant consumption, etc.
• To fit in with HACCP/prerequisite management schedules
• New knowledge (e.g. allergen thresholds)
• New more sensitive sampling device technology
Finally, a validation review would also be required following any cleaning optimisation studies. Optimisation is a process where
cleaning parameters can be manipulated to reduce the cost of the clean (lower temperatures or detergent concentrations) or improve
its efficiency (e.g. reduced rinse times), after the cleaning programme has initially been validated whilst maintaining the validated
objectives and target levels.

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