Cleaning

Validation

Prepared by: Ahmed Elsayed

LinkedIn :
https://www.linkedin.com/in/ahmedel
sayed92/

Email : [email protected]
Cleaning Validation
• Cleaning Validation is a critical process in the pharmaceutical, biotechnology, and
medical device industries to ensure that manufacturing equipment is adequately
cleaned and free from residues that could contaminate future batches.
• Definition:
Cleaning Validation is documented evidence that an approved cleaning
procedure removes residues of the product, cleaning agents, and microbial
contamination to pre-defined acceptable limits.
• Objectives of Cleaning Validation
 Prevent cross-contamination
 Ensure product quality and patient safety
 Comply with regulatory requirements (e.g., FDA, EMA, WHO)
 Maintain GMP (Good Manufacturing Practice)
What Needs to Be Validated in Cleaning Validation?
• Cleaning Validation must cover all product-contact surfaces and cleaning
processes that may directly or indirectly impact product quality or patient
safety. The scope depends on your facility’s processes, product types, and
cleaning method (manual, CIP, or COP).

Area Validation Requirement

Equipment Surface cleanliness, residue removal
Utilities (CIP, WFI, steam) Cleanliness, absence of biofilm or residues
Utensils and cleaning tools No microbial or chemical carryover
Cleaning methods (manual/CIP) Repeatability, effectiveness, coverage
Detergents and agents No toxic or interfering residues
Hidden and difficult areas Swab/rinse accessibility, no residue entrapment (
Dirty/Clean hold times Time limits validated, cleaning effectiveness maintained
Product changeovers Removal of previous product to within acceptable limits
1. Manufacturing Equipment
• These are all product-contact equipment that must be validated for effective cleaning
Common Examples:
Equipment Type Examples Key Risks
Blending/Mixing Ribbon blender, V-blender, tank mixers Powder residues, cross-contamination
Granulation Fluid bed granulator, high-shear mixer Sticky binders, microbial growth
Compression/Encapsulation Tablet press, capsule filler Powder residues, allergens, cross-contamination
Coating Equipment Film coater, pan coater Film coating residues, solvents
Filling Lines Liquid filling machines, nozzles Product residues, microbial contamination
Holding Tanks WIP tanks, storage tanks Product hold-up, difficult-to-clean areas
Packaging Machines Blister packers, tube fillers Product carryover from open systems

Why Validate? To ensure equipment surfaces do not retain residues that contaminate
subsequent products.
2. Cleaning Tools and Utensils
• Even non-product-contact cleaning tools can become a source of contamination.
Examples:
• Brushes, mops, scrapers
• Buckets and cloths
• Disassembled nozzles and spray balls
Risk: If not properly cleaned or sanitized themselves, they can introduce residues or
microbes during cleaning.
3. Product Contact Utilities
Even support utilities that contact the product must be considered.
• CIP (Clean-In-Place) systems
• Water systems (WFI, Purified Water)
• Steam systems (for SIP)
Validate their cleanliness and reliability (e.g., no microbial growth in water loops or
buildup in CIP lines).
• 4. Hard-to-Clean Parts and Hidden Areas
Areas difficult to inspect or reach are more prone to residue retention.
• Dead legs
• Weld seams and gaskets
• Sight glasses
• Spray balls
• Agitator seals
These may require special sampling or enhanced design controls during equipment
qualification.
5. Non-Dedicated Equipment
This is high priority. Equipment used for multiple products must be validated more
rigorously.
Risk of cross-contamination between active pharmaceutical ingredients (APIs), especially
for: Potent APIs, Hormones/steroids, Penicillins, Cytotoxics, Allergens
Use of campaign cleaning, bracketing, or worst-case product selection is typical in these
cases.
• 6. Cleaning Agents and Detergents
Validation must show that:
• Cleaning agents are fully removed
• They don’t interfere with product quality
• Their residues are within safe limits
Residual cleaning agents themselves may be toxic or reactive.

7. Manual vs. Automated Cleaning Methods

• Each cleaning method must be validated:
Type of Cleaning Validation Focus Areas
Manual Cleaning Operator variability, technique, accessibility
CIP Systems Flow patterns, spray coverage, dead-leg validation
COP Systems Contact time, detergent concentration, rinsing

Don’t assume that CIP systems clean everything equally- spray coverage studies may be
required.
 Spray Coverage Studies in Cleaning Validation
 Spray Coverage Studies are a critical part of Cleaning Validation, especially when
automated systems like CIP (Clean-in-Place) or spray balls/nozzles are used. They ensure
that all product-contact surfaces receive adequate contact with the cleaning solution -
no shadowing, blind spots, or “dry zones.”
Purpose
To verify and document that:
• Cleaning agents reach all internal surfaces
• There are no uncleaned zones due to spray pattern limitations
• The design of the equipment (including spray devices) allows complete coverage
👉 Especially important for:
CIP tanks, Fermenters/bioreactors, Vessels with spray balls, Isolators or closed RABS,
Equipment with complex geometry
When Are Spray Coverage Studies Required?
• New CIP systems or equipment installations
• Equipment modifications
• Change in spray ball design or cleaning strategy
• Validation of cleaning coverage for worst-case cleaning scenarios
Study Methodologies
1. Riboflavin Test (Fluorescent Dye Test) – Most Common
A simple, visual method using Riboflavin (Vitamin B2) solution that fluoresces under UV
light.
• Advantages:
• Easy to perform
• Visually clear results
• No special instruments needed
• Steps:
• Prepare solution (typically 0.2–0.5% riboflavin in water).
• Spray coat all internal surfaces of the equipment with the solution.
• Run a CIP cycle (without cleaning agent) using water only.
• Inspect under UV light (365 nm) for any residual fluorescence.
• Document results using photos and coverage maps.
No fluorescence = successful coverage
Typically done during equipment qualification (IQ/OQ)
8. Cleaning Hold Time and Dirty Hold Time
You must validate:
Dirty Hold Time: Maximum time equipment can sit uncleaned without residue becoming
harder to remove.
Clean Hold Time: How long cleaned equipment can be held before it must be re-cleaned
or re-sanitized.
9. Product Changeovers
If your facility manufactures multiple products, validate:
• Changeover cleaning effectiveness
• Worst-case product carryover
• Sequence of changeovers

Especially critical in multi-product or shared facilities.

 Types of Cleaning
1. Manual Cleaning
Performed by operators using physical actions (scrubbing, wiping) with cleaning agents.
• Done with brushes, cloths, or scrapers. Requires SOPs and training.
• Suitable For Small, hard-to-reach equipment; one-off cleanings
• Risks Operator-dependent, inconsistent, harder to validate

• Manual cleaning refers to the process where operators physically clean equipment or
parts by hand, using cleaning tools (brushes, cloths) and cleaning agents (detergents,
solvents).
• Unlike automated systems (CIP/COP), manual cleaning is operator-dependent, and that
makes validation more complex and risk-prone — but still crucial to ensure equipment
cleanliness, product safety, and regulatory compliance.
Objectives of Manual Cleaning Validation
• Prove that the manual cleaning process consistently removes residues of product, detergent, and
microbes.
• Ensure repeatability, despite human variability.
• Define validated parameters and limits for routine cleaning and inspection.
• Document that the process meets GMP and regulatory expectations.
Manual Cleaning Validation Process
1. Develop a Cleaning Procedure (SOP).
Cleaning steps and sequence, required tools (e.g., brushes, scrapers), type and concentration of
cleaning agent, Contact time and rinsing requirements, Inspection points and visual checks, limits
for hold times (dirty & clean).
2. Training and Qualification of Personnel.
Because the outcome depends on human actions, operator training is essential.
• Train all cleaning staff on the procedure
• Qualify them with mock runs and real cleans
• Record training and retraining (especially if deviation occurs)
3. Worst-case Selection
Manual cleaning validation should be based on worst-case scenarios:
• Equipment that’s hardest to clean
• Products that are stickiest, most potent, or least soluble
• Dirtiest allowable condition (max dirty hold time)
• Lowest skill operator (least experienced but trained).
4. Sampling Method
To assess cleanliness, you must collect samples from cleaned surfaces:
Type of Sampling Description When to Use
Swab Sampling Physical swab from specific hard-to-clean locations Preferred for manual cleaning
Rinse Sampling Final rinse collected and analyzed Supplementary, if accessible
Visual Inspection No visible residue is the minimum requirement Always required

Swab sites must represent: Worst-case surfaces (corners, valves, dead ends), product contact
surfaces, operator reach points
5. Analytical Methods
Use validated analytical methods to detect:
• Product residue (e.g., API or excipient)
• Cleaning agent (e.g., alkaline detergent)
• Bioburden/microbial contamination (especially in sterile areas)
Common methods:
• HPLC, TOC, UV, conductivity
• Microbial enumeration (CFU/mL or swab)
6. Acceptance Criteria
Establish scientific and risk-based limits, typically:
• No visible residue
• Quantitative limits, e.g.:
1. Not more than 10 ppm of previous product
2. Not more than 1/1000 of minimum daily dose
3. Not exceeding PDE (Permitted Daily Exposure)
4. Microbial limits (if applicable)
These must be justified, documented, and approved.
Revalidation Triggers
• Manual cleaning should be revalidated when:
• Procedure changes (different tools, agent, steps)
• New product or equipment introduced
• Cleaning failures/deviations
• Changes in personnel or layout
• Change in product classification (e.g., potent APIs)
Validated Parameters (Process Parameters)
These are the controllable elements of the cleaning process that must be defined and
maintained to ensure effectiveness.
1- Cleaning Agent Type : Must be suitable for the residue type (alkaline for organics, acid for scale,
enzyme for proteins).
2- Concentration : too low = ineffective; too high = residue risk (defined range, e.g., 1%).
3- Temperature : affects solubility and detergent action (often 40–70°C depending on agent).
4- Contact Time : time needed for the cleaning agent to act (15–30 min is typical, must be validated)
5- Flow Rate / Pressure : for CIP or spray-based systems to ensure turbulence (≥1.5 m/s for turbulent flow
(ASME BPE standard).
6- Rinse Volume : must be sufficient to remove both product and cleaning agent (Validated
through rinse sample testing)
7- Drying Method and Time : Prevents microbial growth or re-contamination (Air-dry or hot air filtered;
validated per equipment)
8- Dirty Hold Time : max time equipment can sit dirty before cleaning (Defined by study e.g., ≤6 hours)
9- Clean Hold Time : max time cleaned equipment remains before re-cleaning (Based on microbial or
dust risk (e.g., 24–72 hours)
Validated Acceptance Limits (Cleaning Outcome Limits)
These are the residue levels that must not be exceeded after cleaning. They ensure no
carryover and no safety risk.
A. Visual Cleanliness
"No visible residue" is the minimum requirement
Visual inspection is performed under:
• Defined lighting (≥ lux level)
• Fixed distance and angle
• Clean, dry equipment
• Training of inspectors is critical.
Note: Visual inspection is not sufficient alone for validation.
B. Chemical Residue Limits
Product Residues (API or Excipient)
• These are remnants of:
• API – the therapeutic component of the previous product
• Excipient – inactive ingredients (binders, coatings, stabilizers, etc.)
• Other process materials – e.g., lubricants, glidants, flavoring agents.

There are three main approaches:

1- Health-Based Limit (PDE approach)
Uses toxicological data (NOAEL, safety factors) to calculate Permitted Daily Exposure
(PDE).
 Resulting in a MACO (Maximum Allowable Carryover) based on:
• Patient body weight
• Dose of next product
• Equipment surface area
2- Dosage-Based Approach
Based on 1/1000 of the minimum daily dose of the previous product.
Common in older validation protocols, but less scientifically robust than PDE.

3- Analytical Detection Limits

E.g., NMT 10 ppm residue limit or based on TOC/UV/HPLC LOD/LOQ values.
1- Health-Based MACO Calculation (Using PDE)
Step 1: Gather Data
Parameter Value
PDE of previous product 2 mg/day
Batch size of next product 500,000 tablets
Max. daily dose of next product 1000 mg/day

Step 2: MACO Formula (Health-Based)

Step 3: Convert to Equipment Surface Area Limit

Assume shared equipment has surface area = 50,000 cm²

Each swab sample must detect ≤ 20 µg/cm² residue for acceptance.

Step 4: Check Method Suitability (Analytical Method)
Assume your HPLC method LOD = 5 µg/cm², LOQ = 10 µg/cm² → Method is acceptable
(below the MACO limit).

Notes:
• PDE values are usually obtained from toxicological reports or published literature.
• If PDE = 0.5 mg/day → residue limit becomes stricter (quarter of the above).
• EMA and WHO now recommend health-based over dose-based limits.
• Example
Parameter Value
API A (previous product) Atenolol
PDE of API A (from toxicology report) 3 mg/day
Next product (Product B) Paracetamol tablets
Maximum daily dose of Product B 2000 mg/day
Batch size of Product B 1,000,000 tablets (500 mg each)
Total equipment surface area 60,000 cm²

This means: 1.5 grams of Atenolol can remain on the equipment and still be safe for the
next product’s patients.
 Now calculate the residue per cm² allowed on the equipment:

 If swabs are taken over 25 cm²:

So, any swab sample collected from the equipment surface must not exceed 625 µg of API A.
Ensure the Analytical Method is Suitable
Parameter Value
Method LOD (Limit of Detection) 0.5 µg/cm²
Method LOQ (Limit of Quantitation) 1.0 µg/cm²
Limit required 25 µg/cm²

👉 your method is sensitive enough, as LOD and LOQ are well below the MACO limit.
Final Summary :

Item Value
API A PDE 3 mg/day
Next product dose 2000 mg/day
Batch size 1,000,000 tablets
Total MACO 1500 mg
Equipment surface area 60,000 cm²
Acceptance limit (µg/cm²) 25 µg/cm²
Acceptance per swab (25 cm²) 625 µg (0.625 mg)
B. Dose-Based Approach (1/1000 MDD)
➤ Formula

Then scaled based on:

• Example
Let’s say:
• Previous product (Product A): MDD = 500 mg/day
• Next product (Product B): MDD = 1000 mg/day
• Batch size of B = 100,000 g = 100,000,000 mg
• Shared surface area = 20,000 cm²
_______________________________________________________

👉 your residue limit: 250 µg/cm² for Product A in the equipment before making Product B
 Thanks
Prepared by: Ahmed Elsayed

LinkedIn : https://www.linkedin.com/in/ahmedelsayed92/

Email : [email protected]