The document discusses cleaning validation, including definitions, purposes, mechanisms, agents, methods, parameters, groupings, worst case considerations, acceptance criteria, and more. Cleaning validation is defined as the process of removing contaminants from equipment and monitoring such that equipment can be safely reused. It aims to ensure product integrity, prevent cross contamination, and meet regulatory requirements.

1. Know – How of an Effective Cleaning Program
Sambhujyoti Das, Quality Assurance
CLEANING VALIDATION

2. CLEANING VALIDATION…………………... AT A GLANCE
After completing this session we’ll come to know :
 Definition
 Purpose
 Cleaning mechanisms
 Cleaning agents
 Cleaning Methods
 Cleaning parameters
 Cleaning continuum
 Grouping strategies
 Worst Case considerations
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Quality Assurance
 Acceptance criteria
 Sampling Methods
 Analytical Methods
 Hold time studies
 USFDA 483 Citations

3. CLEANING VALIDATION……………… THE DEFINITION
The process of removing contaminants from process
equipment and monitoring the condition of equipment
such that the equipment can be safely used for
subsequent product manufacturing.
Dustin A. Leblanc.
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Quality Assurance

4. CLEANING VALIDATION…………………........... PURPOSE
 Product integrity
Cross contamination
Microbial integrity
Product impurity
Batch integrity
 Equipment reuse
 Regulatory issues
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Quality Assurance

5. CLEANING VALIDATION…………CLEANING MECHANISMS
The chemistry of contaminant removal :
 Solubility
 Wetting
 Emulsification
 Dispersion
 Hydrolysis
 Oxidation
 Physical removal
 Antimicrobial action
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Quality Assurance

6. CLEANING VALIDATION…………CLEANING MECHANISMS
Solubility :
Solubility involves the dissolution of one chemical (the
contaminant) in a liquid solvent.
For example, salts may be soluble in water, and certain organic
actives may be soluble in acetone or methanol.
One of the primary cleaning mechanisms to be considered
during design phase.
Rate of solubility, Insoluble form, Soluble – Insoluble species
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Quality Assurance

7. CLEANING VALIDATION…………CLEANING MECHANISMS
Wetting :
Wetting involves the displacement of one fluid from a solid
surface by another fluid. Wetting can be improved by the
addition of surfactants.
It improve penetration of the cleaning solution into cracks
and crevices, which are usually difficult-to clean locations.
6Courtesy: Validated Cleaning Technologies for Pharmaceutical Manufacturing, D. A. LeBlanc
Quality Assurance

8. CLEANING VALIDATION…………CLEANING MECHANISMS
Emulsification :
Breaking up an insoluble liquid residue into smaller droplets and
then suspending those droplets throughout the water.
Emulsion = Mechanical energy + Surfactants / Polymers.
Emulsions are thermodynamically unstable (say, 5 to 10 mins.).
Redeposition of the cleaned residue back onto the equipment
surfaces.
Agitation should be continued till the time to discharge the
cleaning solution to the drain.
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Quality Assurance

9. CLEANING VALIDATION…………CLEANING MECHANISMS
Dispersion :
Dispersion is similar to emulsification, except that it involves the
wetting and deaggregation of solid particles and then the
subsequent suspension of those particles in water.
More important in dry product manufacturing.
Hydrolysis :
This involves the cleavage of certain bonds in an organic molecule.
The resultant hydrolyzed residues must either be water
soluble or solubilized at the pH of the cleaning solution.
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Quality Assurance

10. CLEANING VALIDATION…………CLEANING MECHANISMS
Oxidation :
This involves the cleavage of various organic bonds, such as
carbon-carbon bonds, by the action of a strong oxidizing agent.
Large Non-polar Mol. Smaller more polar Mol.
Antimicrobial Action :
Mechanisms that may kill organisms but leave behind nonviable
microbial residues.
Special type of mechanism, sterilization, disinfection.
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Quality Assurance

11. CLEANING VALIDATION…………CLEANING MECHANISMS
Physical Removal:
Cleaning by some mechanical force. the objective is to physically
displace the residue.
Pressurized water + Scrubbing
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In real life situation, more than
one cleaning mechanisms are
being used.
Quality Assurance

12. CLEANING VALIDATION……………….CLEANING AGENTS
Cleaning Agents
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Aqueous Cleaning Organic Solvents
Water Surfactants
Chelants
Solvents (miscible)
Acids / Bases
Oxidants
Quality Assurance

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CLEANING VALIDATION……………….CLEANING AGENTS
Organic
Solvents
• Acetone
• Methanol
• Ethyl
Acetate
Surfactants
• SLS
• SDS
• Fatty acid
salts
Chelants
• EDTA
• NTA
• SHMP
Solvents
(miscible)
• Glycol
Ethers
Bases
• NaOH
• KOH
Acids
• Glycolic
Acid
• H3PO4
• Citric Acid
Oxidants
• NaOCl
• Peracetic
Acid
• H2O2
Quality Assurance

14. CLEANING VALIDATION…………….CLEANING METHODS
Automated Cleaning:
o Fixed CIP
o Portable CIP
o Parts Washer
o Ultrasonic
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Manual Cleaning:
Soak
Brush
Wipe
Spray
Extent of automation……………..Extent of disassembly
Quality Assurance

15. CLEANING VALIDATION…………….CLEANING METHODS
Fixed CIP :
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Quality Assurance

16. CLEANING VALIDATION…………….CLEANING METHODS
Portable CIP :
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Quality Assurance

17. CLEANING VALIDATION…………….CLEANING METHODS
Parts Washer :
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Ultrasonic Washer :
Quality Assurance

18. CLEANING VALIDATION…….……CLEANING PARAMETERS
 Time
 Action
 Cleaning chemistry
 Concentration
 Temperature
 Mixing / flow / turbulence
 Water quality
 Rinsing
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Quality Assurance

19. Parameter interactions :
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CLEANING VALIDATION…….……CLEANING PARAMETERS
Time vs Concentration :
Temp. vs Concentration :
Courtesy: Validated Cleaning Technologies for
Pharmaceutical Manufacturing, D. A. LeBlanc
Quality Assurance

20. Parameter interactions :
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CLEANING VALIDATION…….……CLEANING PARAMETERS
Time vs Temperature :
Time (min)
Courtesy: Validated Cleaning Technologies for
Pharmaceutical Manufacturing, D. A. LeBlanc
Quality Assurance

21. CLEANING VALIDATION…….……CLEANING CONTINUUM
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Continuum represent the extremes in the range of operating
differences found within the industry.
The continuum should be used during the initial phases of
defining a cleaning validation program or during new
product development.
Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Automated Cleaning
COP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..CIP
Dedicated Equipment . . . . . . . . . . . . . Non-Dedicated Equipment
Product Contact Surfaces . . . . . . . Non-Product Contact Surfaces
Non-Critical Site . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Critical Site
Minor Equipment . . . . . . . . . . . . . . . . . . . . . . . . Major Equipment
Quality Assurance

22. CLEANING VALIDATION…….……CLEANING CONTINUUM
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Low Risk Drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . High Risk Drugs
Highly Characterized . . . . . . . . . . . . . . . . . . . Poorly Characterized
Sterile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Non-Sterile
Solid Formulations . . . . . . . . . . . . . . . . . . . . . Liquid Formulations
Soluble . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Insoluble
Single Product Facility . . . . . . . . . . . . . . . Multiple Product Facility
Campaigned Production . . . . . . . . . Non-Campaigned Production
Simple Equipment Train . . . . . . . . . . . Complex Equipment Train
Quality Assurance

23. CLEANING VALIDATION…….……GROUPING STRATEGIES
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"Grouping" is the concept of demonstrating that certain elements
of cleaning are of a similar type, and selecting one (or more)
representative object(s) on which to conduct the Cleaning
Validation (Cleaning Process Qualification).
Product grouping :
 Same manufacturing equipments being used.
 Same cleaning SOPs being followed.
 Similar formulations.
 Similar risk / therapeutic group.
Equipment grouping, Cleaning method grouping, Cleaning
agent grouping, …………….., etc.
Quality Assurance

24. CLEANING VALIDATION…….……GROUPING STRATEGIES
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Sr. No. Name of product Formulation
Cleaning
methods
Equipment
train
Risk /
Therap. class
1 Product A Tablet (FC) Method 1 Train A General
2 Product B Tablet Method 1 Train B General
3 Product C Parenteral Method 2 Train C Cytotoxic
4 Product D Tablet Method 3 Train B General
5 Product E Tablet (EC) Method 4 Train A General
6 Product F Parenteral Method 2 Train C Cytotoxic
7 Product G Tablet (FC) Method 1 Train A Cytotoxic
8 Product H Tablet Method 3 Train B General
9 Product I Tablet (EC) Method 4 Train A General
10 Product J Parenteral Method 2 Train C Cytotoxic
All products in a facility (hypothetical):
Quality Assurance

25. CLEANING VALIDATION…….……GROUPING STRATEGIES
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Sr. No. Name of product Formulation
Cleaning
methods
Equipment
train
Risk /
Therap. class
1 Product A Tablet (FC) Method 1 Train A General
2 Product B Tablet Method 1 Train B General
3 Product C Parenteral Method 2 Train C Cytotoxic
4 Product D Tablet Method 3 Train B General
5 Product E Tablet (EC) Method 4 Train A General
6 Product F Parenteral Method 2 Train C Cytotoxic
7 Product G Tablet (FC) Method 1 Train A Cytotoxic
8 Product H Tablet Method 3 Train B General
9 Product I Tablet (EC) Method 4 Train A General
10 Product J Parenteral Method 2 Train C Cytotoxic
Before Grouping :
Quality Assurance

26. CLEANING VALIDATION…….……GROUPING STRATEGIES
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Sr. No. Name of product Formulation
Cleaning
methods
Equipment
train
Risk /
Therap. class
1 Product A Tablet (FC) Method 1 Train A General
2 Product B Tablet Method 1 Train B General
3 Product G Tablet (FC) Method 1 Train A Cytotoxic
4 Product C Parenteral Method 2 Train C Cytotoxic
5 Product F Parenteral Method 2 Train C Cytotoxic
6 Product J Parenteral Method 2 Train C Cytotoxic
7 Product D Tablet Method 3 Train B General
8 Product H Tablet Method 3 Train B General
9 Product E Tablet (EC) Method 4 Train A General
10 Product I Tablet (EC) Method 4 Train A General
After Grouping :
Quality Assurance

27. CLEANING VALIDATION…..WORST CASE CONSIDERATIONS
Once the product groups have been established, the next step is to
determine the so-called “worst case” representative of each group.
It is that member(s) who shows the highest challenge on
cleaning program.
Worst case product : Toxicity / solubility / highly characterized
/ difficult to clean ingredients.
Worst case eq. train : Longest train.
Worst case equipment : Larger size equipment (identical design).
Worst case acc. criteria: Stringent acceptance criteria.
Hold time studies : Longest possible duration.
Campaign Mfg. : Highest possible nos. of batches.
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Quality Assurance

28. CLEANING VALIDATION…..WORST CASE CONSIDERATIONS
There is no ‘hard & fast’ rule on worst case selection.
A good logic and science should always be used.
Grouping and worst case selection help to demonstrate
cleaning method robustness.
It smartly reduces the load from cleaning validation
program.
These philosophies should always be verified against the
actual capability of cleaning program.
The ultimate ‘cost – benefit’ ratio should be evaluated.
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Quality Assurance

29. CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
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How clean is clean ?
What are the bases of defining limits ?
What are the impacts of after cleaned residue ?
Human Drug CGMP Notes, 9:2, 2Q 2001 :
“Should equipment be as clean as the best possible method
of residue detection or quantification?”
Answer: “No,……absolute cleanliness is neither valuable
nor feasible…. It should be as clean as can be reasonably be
achieved, to a residue limit that is medically safe and that
causes no product quality concerns…………….”
Quality Assurance

30. Three criteria :
 It should be scientifically justifiable.
 Pacifically achievable.
 Methodically verifiable.
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Possible types of limits :
 Visual
 Chemical
 Microbiological
 Endotoxin
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Quality Assurance

31. Visual clean criteria :
GMPs require inspection for visual cleanness before manufacture.
Key items to consider :
o Angle of view
o Distance from equipment surface
o Lighting conditions
o Viewer’s knowledge
o Surface usually must be dry
Visual aids :
Additional lighting / Magnifying glass / Mirror / Fiber-optic
scope / UV light
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Quality Assurance

32. Application for visual limits :
A typical visual limit is NLT 4 μg / cm2.
“Visually clean” may not be enough by itself
 Potent drugs
 Microbial contamination
 Endotoxin
More suitable method for non-potent drug products and APIs.
PIC/S advocates spiked coupon study for determination of
visual inspection limits (and for training of inspectors).
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Quality Assurance

33. Chemical residue limits (Therapeutically or Toxicologically
safe criteria) :
 Therapeutic dose based criteria
Most suitable for drug product (finished product)
manufacturing facility.
 Toxicological criteria
Most suitable for active drug (API) manufacturing facility.
Where cleaning agents are used (other than water).
 10 PPM criteria
CGMP requirement widely applicable.
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Quality Assurance

34. Therapeutic dose based criteria :
Based on the assumption that 1/1000 part of therapeutic dose
does not have any clinical impact on human (animal) body.
Determination of MAC (Maximum Allowable Carryover) of
Product A (Previous) to Product B (Next)
SRDD (A) × BS (B) × SF
MAC = (unit of mass)
LRDD (B)
Where, SRDD = Smallest Recommended Daily Dose (Product A – ACTIVE CONTENT),
BS = batch size (Product B), SF = safety factor and LDD and LRDD = Largest
Recommended Daily Dose (Product B – DRUG PRODUCT)
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1
Quality Assurance

35. Therapeutic dose based criteria :
Determination of Surface contamination (Shared Equipment)
MAC
L1 = (mass / surface area)
SESA
Where, SESA = Shared Equipment Surface Area (for both products)
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 2
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 3
Therapeutic dose based criteria :
Determination of Sampled residue (for swab sample)
L2 = L1 × Swab Area (mass / swab)
 SRDD value represents the ACTIVE drug content only.
e.g. 5 mg or 10 mg, the dose strength.
 LRDD value represents the mass or volume of entire dose.
e.g. 250 mg or 20 mL (drug + excipients).
 Convert similar items into similar convenient unit of measure.
Quality Assurance

37. Safety Factors :
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Approach Approach Typically Applicable To
0.1 to 0.01 Topical products
0.01 to 0.001 Oral products
0.001 to 0.0001 Parenterals, opthalmic products
0.0001 to 0.00001 Research, investigational products
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1
Therapeutic dose based criteria (an example) :
Determination of Maximum Allowable Carryover
10 mg × 150 kg × 0.001 × 1000000
(250 mg × 3)
= 2000 mg (MAC value)
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39. 38
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 2
Therapeutic dose based criteria (an example) :
Determination of Surface contamination level
2000 mg
3170 cm2
= 0.63 mg / cm2 (L1 value)
Quality Assurance

40. 39
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 3
Therapeutic dose based criteria (an example) :
Determination of Swab residue
0.63 mg / cm2 × 25 cm2
= 15.75 mg / swab (L2 value)
Quality Assurance

41. Toxicological criteria :
Based on the toxicological information available in Material Safety
Data Sheets.
Determination of NOEL (No Observed Effect Level)
NOEL = LD50 × Emperical Factor
(unit of mass/kg of body weight)
Where, LD50 = lethal dose for 50% of animal population in study (mg/kg/day),
Emperical Factor = derived from animal model developed by Layton, et.al : 0.001*
* Used by expert panel of WHO (10-3).
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1A
Quality Assurance

42. Toxicological criteria :
Determination of ADI (Acceptable Daily Intake)
ADI = NOEL × AAW × SF
(unit of mass)
Where, AAW = average adult weight : 70 kg,
SF = safety factor (0.01)
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1B
 Consider average body weight of child where there is any
pediatric dose available.
 Use LD50 value of mice.
Quality Assurance

43. Toxicological criteria :
Determination of MAC (Maximum Allowable Carryover)
ADI × BS
MAC =
LRDD (any next product) (unit of mass)
Then use and to derive final swab residue
limit.
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1C
Step 2 Step 3
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1A
Toxicological criteria (an example) :
Determination of NOEL
(1750 mg /kg/day) × 0.001 = 1.75 mg/kg
(NOEL value)
Determination of ADI
(1.75 mg/kg) × 70 kg × 0.01 = 1.225 mg
(ADI value)
Step 1B
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1C
Toxicological criteria (an example) :
Determination of MAC
1.225 mg × 150 kg × 1000000
(250 mg × 3)
= 245000 mg
The final Swab residue (L2) :
245000 mg × 25 cm2
3170 cm2
= 1932 mg/swab
Quality Assurance

46. 10 PPM criteria :
Based on the hypothesis that 10 parts of previous product is
therapeutically ineffective if presents in million parts of next
product.
Determination of MAC
10 × BS
MAC = (unit of mass)
1000000
Where, BS = batch size (smallest available batch size)
Then use and to derive final swab residue
limit. 45
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1
Step 3Step 2
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47. 46
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Step 1
10 PPM criteria (an example) :
Determination of MAC
10 × 150 kg × 1000000
MAC = = 1500 mg
1000000
The final Swab residue (L2) :
1500 mg × 25 cm2
3170 cm2
= 11.83 mg/swab
Quality Assurance

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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
The most stringent acceptance criteria shall be chosen for
cleaning validation study (The worst case approach).
11.8315.75 1932
mg / swab
In real life cases, therapeutic or 10 PPM criteria become final
acceptance criterion for cleaning validation.
Quality Assurance

49. Microbiological criteria :
 Internal specifications
 Official specifications: e.g. USP <1111>, “Microbial
Examination of nonsterile Products: Acceptance criteria for
Pharmaceutical Preparations and Substances for Pharmaceutical
Use”
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Adminstration
route
Total aerobic count
(cfu/g or cfu/mL)
Total combined
yeasts/molds count
(cfu/g or cfu/mL)
Nonaqueous oral 103 102
Aqueous oral 102 10
Most topicals 102 10
Quality Assurance

50. Microbiological criteria :
 Environmental specifications: EU GMP, Annex – 1,
“Recommended limits for microbiological monitoring of clean
areas during operation”
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Grade Contact plates (diam. 55 mm), cfu/plate
A < 1
B 5
C 25
D 50
i.e. recommended limit for microbial contamination in grade D
area is : 50/{π × (5.5/2)2}= 2.10 cfu/cm2
Quality Assurance

51. Microbiological criteria from internal specifications:
 Driven by SOP.
 Must be backed up by justifiable scientific rationale.
Microbiological criteria from official specifications:
Spec. limit × factor × Wt. product
SESA
An example:
1000 cfu/g × 0.1 × 5 kg × 103
3170 cm2
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
× swab area
× 25 cm2 = 3943 cfu/swab
Quality Assurance

52. Microbiological criteria from environmental specifications:
50/{π × (5.5/2)2} × swab area
An example:
2.10 cfu/cm2 × 25 cm2 = 52 cfu/swab
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CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Quality Assurance

53. Determining acceptance criteria with more than one next
products (The Matrix approach):
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
NEXT PRODUCT Prod. A Prod. B Prod. C Prod. D Prod. E
(kg) B. Size 200.0 75.0 100.0 150.0 355.5
(cm2) S. Area 4525 3960 4015 3770 4008
(mg) SRDD LRDD GENERAL SOLID FACILITY
Product A 10.0 450.0 10.5 13.8 22.1 49.3
Product B 1.0 320.0 3.4 1.9 3.1 6.9
Product C 25.0 600.0 46.0 19.7 41.4 92.4
Product D 5.0 300.0 18.4 7.9 10.4 36.9
Product E 125.0 800.0 172.6 74.0 97.3 155.4
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PREVIOUS
PRODUCT
Quality Assurance

54. The sampling procedure refers to the method of collecting the
residues from the surface so that they can be measured.
CLEANING VALIDATION…………......SAMPLING METHODS
Types Advantages Limitations
Swabs & Wipes
Dissolves & physically removes
sample, adaptable to wide variety
of area
May introduce fibers,
technique dependent, hard-to-
reach areas
Rinse
Easy, quick, non-intrusive, large
surface area
Limited information about
actual surface cleanliness
Coupon
Non-technique dependent, reduces
variability in recovery
Invasive, might interfere with
cleaning process
Placebo
Placebo contacts the same surfaces
as the product
Difficult to determine
recovery
Direct Surface Rapid, non-invasive, economical
Some techniques not widely
developed
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Quality Assurance

55.  Swab sampling techniques:
(1) One of the most widely used technique for chemical and
microbial sampling.
(2) Swabs are being wet with solvent aiding solubilization and
physical removal of surface residues.
(3) Results are technique dependent.
CLEANING VALIDATION…………......SAMPLING METHODS
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Microbial swab (sterile) Chemical swabs (Texwipe) Cotton wipes
Quality Assurance

56.  Swab sampling techniques:
(5) Generally 1 swab sample per location is adequate.
(6) Multiple swabs can be taken to improve surface recovery.
(7) Typical swabbed per site varies from 25 cm2 to 100 cm2. There
is no “magic” number.
(8) PTFE (chemically inert) templates may be used for accurate
swabbing area.
(9) “Difficult to clean” equipment surfaces
shall be identified and sampled.
(10) Representative surfaces of different
materials (MOCs) should be sampled.
CLEANING VALIDATION…………......SAMPLING METHODS
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5 cm
5 cm
2.5
cm
10
cm
Swab area
templates
Quality Assurance

57.  Swab sampling techniques:
(11)Wiping should be unidirectional at a time. Parallel strokes
should be employed to cover entire swab area.
CLEANING VALIDATION…………......SAMPLING METHODS
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Courtesy: Validated Cleaning Technologies for Pharmaceutical Manufacturing, D. A. LeBlanc
Quality Assurance

58.  Swab sampling techniques:
Example of “Difficult to clean” locations of an RMG:
CLEANING VALIDATION…………......SAMPLING METHODS
57
Courtesy: Rapid mixer granulator, Kevin.
The design aspect of the equipment should be considered to
identify “difficult to clean” locations.
Quality Assurance

59.  Rinse sampling techniques:
Rinse sampling involves using a liquid to cover the surfaces to be
sampled.
(1) One of the easy and widely used sampling method.
(2) Most preferable liquid for rinsing is water.
(3) The rinse volume is an important factor that has to be
determined.
Rinse volume α (1/Residue conc. in rinse sample)
(4) Forced rinsing is advisable for collection of less soluble
residues.
CLEANING VALIDATION…………......SAMPLING METHODS
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Quality Assurance

60.  Determination rinse volume:
(1) Variability in magnitudes of surface areas gives rise of variable
residue concentrations in rinse samples (fixed rinse volume).
(2) Variable acceptance criteria for a single product creates
confusion.
(3) It is a good idea to chose variable rinse volumes to keep
constant residue concentration in rinse samples (fixed
acceptance criteria).
Formula :
L1 × ESA
Rinse vol. for Equipment A =
Anticipated rinse conc.
CLEANING VALIDATION…………......SAMPLING METHODS
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Quality Assurance

61.  Determination rinse volume:
Example :
0.63 mg / cm2 × 1760 cm2
Rinse vol. for Equipment A =
10 μg / mL
= 110.9 L (considering mg/L = PPM)
0.63 mg / cm2 × 810 cm2
Rinse vol. for Equipment B =
10 μg / mL
= 51.0 L
CLEANING VALIDATION…………......SAMPLING METHODS
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Quality Assurance

62.  Specific vs non-specific methods:
(1) A non-specific assay may detect a variety of residues.
(2) A specific assay may quantify any anticipated residue.
(3) It is essential to correlate the results from a specific method to
the results from other non-specific methods that might be
used for routine monitoring of cleaning effectiveness.
CLEANING VALIDATION…………ANALYTICAL METHODS
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HPLC pH meter
Quality Assurance

63. CLEANING VALIDATION…………ANALYTICAL METHODS
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Specific Test Methods Non-Specific Test Methods
UV/Visible Spectrophotometry
Near Infrared Spectrophotometry
(NIR)
High Performance Liquid
Chromatography (HPLC)
Mid Infrared Spectrophotometry (MIR)
Atomic Absorption
Capillary Zone Electrophoresis
Enzyme Linked Immunosorbant Assay
(ELISA)
Total Organic Carbon (TOC)
pH
Titration
Conductivity
Gravimetric
Quality Assurance

64. The analytical methods used for testing cleaning samples
must be validated for [ICH Q2 (R1)]:
 Limit of Detection (LOD)
 Limit of Quantification (LOQ)
 Specificity
 Accuracy
 Repeatability
 Precision
 Range
 Linearity
 Recovery
CLEANING VALIDATION…………ANALYTICAL METHODS
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Quality Assurance

65.  The analytical method used for evaluation of cleaning sample is
different that used for product assay.
 If the target limit in the analytical sample were 5.2 μg / mL,
and a method was only able to detect down to 7.0 μg / mL,
that method would not be useful for cleaning validation
purposes.
 The target value should be within the linearity range of the
specific method.
What if the calculated acceptance value is
less than the detectable level of an
analytical method?
There may be two options available……….
CLEANING VALIDATION…………ANALYTICAL METHODS
64
Quality Assurance

66. Choose more efficient analytical method !
Example:
Derived acceptance limit = NMT 4.0 μg / mL
Analytical LOQ = 5.5 μg / mL
Analytical Method = UV/Visible Spectrophotometry
New method adopted = Ion mobility spectrometry
New LOQ = 2.0 μg / mL
CLEANING VALIDATION…………ANALYTICAL METHODS
65
Quality Assurance

67. Increase the sampling area to achieve at least LOQ
value!
Example:
Derived acceptance limit = NMT 4.0 μg / mL
Analytical LOQ = 5.5 μg / mL
Swab area = 25 cm2
CLEANING VALIDATION…………ANALYTICAL METHODS
66
Revised swab area =
25 cm2
4.0 μg / mL
× 5.5 μg / mL
= 35 cm2 (7 cm × 5 cm)
Quality Assurance

68. Recovery studies :
Procedure :
o Spike coupon with known amount
o Allow to dry
o Remove in swab or simulated rinse procedure
o For swab, desorb
o Analyze sample
o Compare to expected 100% value
This is done at surface acceptance (or below) limit.
CLEANING VALIDATION…………ANALYTICAL METHODS
67
Quality Assurance

69. Swab recovery schematic :
CLEANING VALIDATION…………ANALYTICAL METHODS
68
1. Spike control diluent directly
Control
B μg/mL
Control
C μg/mL
Standard
solution
A μg/mL 2a. Spike
coupon
2b. Swab
coupon
2c. Extract swab
Quality Assurance

70. Recovery calculation 1 (Spiked against Standard):
(C μg/mL) × (mL)
% Recovery = × 100
(A μg/mL) × (mL)
CLEANING VALIDATION…………ANALYTICAL METHODS
69
 Recovery depends on spiked standard of known concentration.
 Disorbing solvent may be of any volume (mL).
 Recovery depends on material surface, sampling method and
some what on analytical method.
Quality Assurance

71. Recovery calculation 2 (Spiked against Positive control) :
(C μg/mL) × (mL)
% Recovery = × 100
(B μg/mL) × (mL)
CLEANING VALIDATION…………ANALYTICAL METHODS
70
 More useful if defined standard is not readily available.
Swab recover study with multiple analysts :
 Usually 3 replicates by one sampler.
 Use lowest value of any one run.
Quality Assurance

72. Rinse recovery schematic :
CLEANING VALIDATION…………ANALYTICAL METHODS
71
Pipette with
rinse solution
(known volume)
Spiked
coupon
Collection beaker
Spike bottom
of SS beaker
Lab sheker
Case 1 Case 2
Quality Assurance

73. Minimum acceptable recovery:
 Specify in cleaning validation master plan or master protocol.
 Minimum swab recovery of 70 % - 80 %.
 Minimum rinse recovery of 50 %.
 Carry out recovery study for different material surfaces
(Material Of Constructions).
 Chose right wetting solvent (soluble) and absorbent swab
material to improve recovery.
 May allow <50 % recovery with written justification.
CLEANING VALIDATION…………ANALYTICAL METHODS
72
Quality Assurance

74. o DEHT = Max. allowed time, between end of usage and
employing cleaning
o CEHT = Max. allowed time, between end of cleaning and
further usage
CLEANING VALIDATION………………………HOLD TIMES
73
Cleaning Hold Time studies
Cleaned Equipment
Hold Time (CEHT)
Dirty Equipment
Hold Time (DEHT)
Quality Assurance

75. Dirty equipment hold time study (DEHT) :
 Soils may become more difficult to clean over time.
 Maximum DEHT should be in SOPs.
 Maximum time shall be set in conjunction with production.
 Representative / worst case product can be selected for study.
 Equipments support wet processing can be selected.
 If extra cleaning is desirable, then it should be in SOP.
 May be expressed in days but preferably by hours.
 Three runs at maximum time……..safe harbor.
74
CLEANING VALIDATION………………………HOLD TIMES
Quality Assurance

76. Dirty equipment hold time study (DEHT) :
Method
 Carry out microbiological sampling at 24 hr., 48 hr., 36 hr., …...
from the dirty equipments.
 Clean the equipments as per SOPs.
 Carry out chemical sampling after cleaning.
 Compile all results (chemical and microbial).
 Successful results shall standardize the maximum DEHT.
 Failure of any results shall reduce the max. DEHT followed by
another 3 verification runs.
75
CLEANING VALIDATION………………………HOLD TIMES
Quality Assurance

77. Cleaned equipment hold time study (CEHT) :
 Microbiological evaluation is the key focus area.
 Maximum CEHT should be in SOPs.
 Representative / worst case product can be selected for study.
 Vitamins, nutritional supplements, product containing Starch or
Gelatin may represent worst cases.
 Avoid conducting study on antibiotic or antimicrobial products.
 Three runs at maximum time……..safe harbor.
 Protection during storage of cleaned equipments should be as
per SOPs.
76
CLEANING VALIDATION………………………HOLD TIMES
Quality Assurance

78. 77
Cleaned equipment hold time study (CEHT) :
Method
 Clean the equipments as per SOPs.
 Store under protection (as per routine procedure).
 Carry out microbiological sampling at 24 hr., 48 hr., 36 hr., …...
 Verify the results against limit (less than validation limit).
 Successful results shall standardize the maximum CEHT.
 Failure of any results shall reduce the max. CEHT followed by
another 3 verification runs.
 Do not set max. CEHT on “until failure” basis.
CLEANING VALIDATION………………………HOLD TIMES
Quality Assurance

79. 78
Campaign hold study (CHS) :
 Cleaning after production of definite number consecutive
batches.
 Negotiate with production related to number of batches.
 Simulate max. anticipated hours of campaign production.
 Cumulative deposition of residues may accelerate product
degredation.
 Perform cleaning and sampling at the end of campaign.
 Max. CHS (no. of batches + time) should be in SOPs.
 Batch to batch or lot to lot cleaning is advisable.
 More suitable for dedicated product equipments.
CLEANING VALIDATION………………………HOLD TIMES
Quality Assurance

80. CLEANING VALIDATION……………....ALL ASPECTS OF CV
79
Courtesy: Biopharm international
Quality Assurance

81. CLEANING VALIDATION…………USFDA 483 CITATIONS
Cleaning Parameters
“….. cleaning of ….. has not been validated, nor is the
spray temperature, volume or time defined.”
80
Quality Assurance

82. CLEANING VALIDATION…………USFDA 483 CITATIONS
Time of Cleaning
“Equipment cleaning is performed on a “clean until
clean” basis. There has been no determination of the
number of cleanings required to ensure the
cleanliness of the equipment.”
81
Quality Assurance

83. CLEANING VALIDATION…………USFDA 483 CITATIONS
Manual Cleaning
“Hands on training for equipment cleaning operations
is not provided and there is no program in place to
assure cleaning consistency between operators.”
82
Quality Assurance

84. CLEANING VALIDATION…………USFDA 483 CITATIONS
Cleaning Log
“There is no assurance that cleaning is conducted as
stated in their SOPs…… There are no cleaning
logs to indicate that this has been done.”
83
Quality Assurance

85. CLEANING VALIDATION…………USFDA 483 CITATIONS
Poor Cleaning
“….. we observed foreign material on the filter
grates….. Daily cleaning as per SOP…..failed to
remove the material. End of process cleaning as per
SOP…..failed to remove the material. Weekly
cleaning as per SOP……failed to remove the
material.”
84
Quality Assurance

86. CLEANING VALIDATION…………USFDA 483 CITATIONS
Detergent Concentration
“Detergent is dispensed into the ……Stopper Washer
reservoir every third cycle. No data has been
collected to determine the detergent concentration
each cycle…..”
85
Quality Assurance

87. CLEANING VALIDATION…………USFDA 483 CITATIONS
Cleaning Agent Labeling
“The firm’s control over IPA 100% used to clean
equipment in production is inadequate. …..bottles
are not labeled…..with date, expiration or who
dispensed, ….no scientific information….which
would establish an expiration date…..”
86
Quality Assurance

88. CLEANING VALIDATION…………USFDA 483 CITATIONS
Acceptance Limit
“….. acceptance criteria….shall not exceed…..μg/cm2.
There is no data to justify this limit.”
87
Quality Assurance

89. CLEANING VALIDATION…………USFDA 483 CITATIONS
Sampling Locations
“Swabbing was performed on general contact areas
without taking into consideration area such as edges
and crevices.”
88
Quality Assurance

90. CLEANING VALIDATION…………USFDA 483 CITATIONS
Sampling Locations
“Exact / precise swab locations are not identified.”
89
Quality Assurance

91. CLEANING VALIDATION…………USFDA 483 CITATIONS
Swab Sampling
“Swab samples collected…..from different locations
from each piece of equipment are combined into
one sample and tested such.”
90
Quality Assurance

92. CLEANING VALIDATION…………USFDA 483 CITATIONS
Rinse Sampling
“….firm’s validation…..is inadequate in that the rinse
solutions were not analyzed for the presence of the
active ingredient residues that might be present.”
91
Quality Assurance

93. CLEANING VALIDATION…………USFDA 483 CITATIONS
Rinse Sampling
“There is not an exact sampling procedure for the
collection of rinse water samples which takes into
account the surface area involved, time of
contact….., volume of rinse, and temperature of
rinse, along with a formula to calculate the amount
of possible contamination based on analysis.”
92
Quality Assurance

94. CLEANING VALIDATION…………USFDA 483 CITATIONS
Dirty Equipment Hold Time
“No time frames / limitations have been established
for production equipment from end of use to start
cleaning.”
93
Quality Assurance

95. CLEANING VALIDATION…………USFDA 483 CITATIONS
Cleaned Equipment Hold Time
“A time limit for the length of time allowed between
cleaning and the use of the manufacturing
equipment…..has not been established.”
94
Quality Assurance

96. CLEANING VALIDATION…………USFDA 483 CITATIONS
Recovery Studies
“Equipment cleaning validation studies for…..did not
include…..present recovery studies on rinse
samples.”
95
Quality Assurance

97. CLEANING VALIDATION…………USFDA 483 CITATIONS
Recovery Studies
“…..each drug’s recovery test was performed only
once, therefore there is no data to show
reproducibility.”
96
Quality Assurance

98. CLEANING VALIDATION…………USFDA 483 CITATIONS
Recovery Studies
“Your firm is using the average of recovery results
(from different amounts of spiked solution) instead
of the worst case result. Using a value that
represents the average does not ensure that
contamination is not higher than calculated.”
97
Quality Assurance

99. CLEANING VALIDATION…………USFDA 483 CITATIONS
Change Control
“The SOP has been revised twice. No review was
performed to determine if a re-validation was
necessary for the changes……”
98
Quality Assurance

100. CLEANING VALIDATION……………................................???
Quality Assurance

101. CLEANING VALIDATION……...SOURCES OF INFORMATION
 “Guide to Inspections Validation of Cleaning Processes”,
Inspection note by FDA (US).
 “Recommendation on VMP, IQ and OQ, non-sterile process
validation and cleaning validation”, (PIC/S).
 “GMP guide for API”, (ICH, Q7).
 “Guidance on Cleaning Validation”, Health Canada.
Technical sources :
 Points to Consider for Cleaning Validation, PDA 29.
 Points to Consider for Biotechnology Cleaning Validation,
PDA 49
Sambhujyoti Das, Quality Assurance
100

102. CLEANING VALIDATION