CLEANING VALIDATION

Know – How of an Effective Cleaning Program

Sambhujyoti Das, Quality Assurance

CLEANING VALIDATION…………………... AT A GLANCE

After completing this session we’ll come to know :

 Definition  Acceptance criteria

 Purpose  Sampling Methods
 Cleaning mechanisms  Analytical Methods
 Hold time studies
 Cleaning agents  USFDA 483 Citations
 Cleaning Methods
 Cleaning parameters
 Cleaning continuum
 Grouping strategies
 Worst Case considerations
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Quality Assurance
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.

Quality Assurance
CLEANING VALIDATION…………………........... PURPOSE

 Product integrity

Cross contamination
Microbial integrity
Product impurity
Batch integrity

 Equipment reuse

 Regulatory issues

Quality Assurance
CLEANING VALIDATION…………CLEANING MECHANISMS

The chemistry of contaminant removal :

 Solubility

 Wetting

 Emulsification

 Dispersion

 Hydrolysis

 Oxidation

 Physical removal

 Antimicrobial action

Quality Assurance
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

Quality Assurance
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.

Courtesy: Validated Cleaning Technologies for Pharmaceutical Manufacturing, D. A. LeBlanc

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Quality Assurance
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
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
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.

Quality Assurance
CLEANING VALIDATION…………CLEANING MECHANISMS

Physical Removal:
Cleaning by some mechanical force. the objective is to physically
displace the residue.
Pressurized water + Scrubbing

In real life situation, more than

one cleaning mechanisms are
being used.

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Quality Assurance
CLEANING VALIDATION……………….CLEANING AGENTS

Cleaning Agents

Aqueous Cleaning Organic Solvents

Water Surfactants

Chelants

Solvents (miscible)

Acids / Bases
Oxidants
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Quality Assurance
CLEANING VALIDATION……………….CLEANING AGENTS
Organic Solvents
Solvents Surfactants Chelants
(miscible)
• Acetone • SLS • EDTA • Glycol
• Methanol • SDS • NTA Ethers
• Ethyl • Fatty acid • SHMP
Acetate salts

Bases Acids Oxidants

• NaOH • Glycolic • NaOCl

• KOH Acid • Peracetic
• H3PO4 Acid
• Citric Acid • H2O2
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Quality Assurance
CLEANING VALIDATION…………….CLEANING METHODS

Automated Cleaning: Manual Cleaning:

o Fixed CIP Soak
o Portable CIP Brush
o Parts Washer Wipe
o Ultrasonic Spray

Extent of automation……………..Extent of disassembly

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Quality Assurance
CLEANING VALIDATION…………….CLEANING METHODS

Fixed CIP :

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Quality Assurance
CLEANING VALIDATION…………….CLEANING METHODS

Portable CIP :

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Quality Assurance
CLEANING VALIDATION…………….CLEANING METHODS

Parts Washer : Ultrasonic Washer :

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Quality Assurance
CLEANING VALIDATION…….……CLEANING PARAMETERS

 Time
 Action
 Cleaning chemistry
 Concentration
 Temperature
 Mixing / flow / turbulence
 Water quality
 Rinsing

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Quality Assurance
CLEANING VALIDATION…….……CLEANING PARAMETERS
Parameter interactions :
Time vs Concentration :

Temp. vs Concentration :

Courtesy: Validated Cleaning Technologies for

Pharmaceutical Manufacturing, D. A. LeBlanc

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Quality Assurance
CLEANING VALIDATION…….……CLEANING PARAMETERS
Parameter interactions :
Time vs Temperature :

Time (min)

Courtesy: Validated Cleaning Technologies for

Pharmaceutical Manufacturing, D. A. LeBlanc

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Quality Assurance
CLEANING VALIDATION…….……CLEANING CONTINUUM

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

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Quality Assurance
CLEANING VALIDATION…….……CLEANING CONTINUUM

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

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Quality Assurance
CLEANING VALIDATION…….……GROUPING STRATEGIES

"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.
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Quality Assurance
CLEANING VALIDATION…….……GROUPING STRATEGIES
All products in a facility (hypothetical):
Cleaning Equipment Risk /
Sr. No. Name of product Formulation
methods train 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

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Quality Assurance
CLEANING VALIDATION…….……GROUPING STRATEGIES
Before Grouping :
Cleaning Equipment Risk /
Sr. No. Name of product Formulation
methods train 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

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Quality Assurance
CLEANING VALIDATION…….……GROUPING STRATEGIES
After Grouping :
Cleaning Equipment Risk /
Sr. No. Name of product Formulation
methods train 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

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Quality Assurance
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
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
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

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…………….”
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

Three criteria :
 It should be scientifically justifiable.
 Pacifically achievable.
 Methodically verifiable.

Possible types of limits :

 Visual
 Chemical
 Microbiological
 Endotoxin

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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
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.
Step 1
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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Therapeutic dose based criteria :
Step 2
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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Therapeutic dose based criteria :
Step 3
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.

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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Safety Factors :
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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

Therapeutic dose based criteria (an example) :

Step 1
Determination of Maximum Allowable Carryover

10 mg × 150 kg × 0.001 × 1000000

(250 mg × 3)

= 2000 mg (MAC value)

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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

Therapeutic dose based criteria (an example) :

Step 2
Determination of Surface contamination level

2000 mg
3170 cm2

= 0.63 mg / cm2 (L1 value)

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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

Therapeutic dose based criteria (an example) :

Step 3
Determination of Swab residue

0.63 mg / cm2 × 25 cm2

= 15.75 mg / swab (L2 value)

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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Toxicological criteria :
Based on the toxicological information available in Material Safety
Data Sheets.
Step 1A
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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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Toxicological criteria :
Step 1B
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)

 Consider average body weight of child where there is any

pediatric dose available.
 Use LD50 value of mice.
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Toxicological criteria :
Step 1C
Determination of MAC (Maximum Allowable Carryover)

ADI × BS
MAC =
LRDD (any next product) (unit of mass)

Then use Step 2 and Step 3 to derive final swab residue

limit.

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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

Toxicological criteria (an example) :

Step 1A
Determination of NOEL

(1750 mg /kg/day) × 0.001 = 1.75 mg/kg

(NOEL value)
Step 1B

Determination of ADI

(1.75 mg/kg) × 70 kg × 0.01 = 1.225 mg

(ADI value)
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

Toxicological criteria (an example) :

Step 1C
Determination of MAC

1.225 mg × 150 kg × 1000000

(250 mg × 3)

= 245000 mg
The final Swab residue (L2) :
245000 mg × 25 cm2
= 1932 mg/swab
3170 cm2
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
10 PPM criteria :
Based on the hypothesis that 10 parts of previous product is
therapeutically ineffective if presents in million parts of next
product.
Step 1
Determination of MAC
10 × BS
MAC = (unit of mass)
1000000
Where, BS = batch size (smallest available batch size)

Then use Step 2 and Step 3 to derive final swab residue

limit. 45

Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

10 PPM criteria (an example) :

Step 1
Determination of MAC

10 × 150 kg × 1000000
MAC = = 1500 mg
1000000

The final Swab residue (L2) :

1500 mg × 25 cm2
= 11.83 mg/swab
3170 cm2
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA

The most stringent acceptance criteria shall be chosen for

cleaning validation study (The worst case approach).

15.75 11.83
1932
mg / swab

In real life cases, therapeutic or 10 PPM criteria become final

acceptance criterion for cleaning validation. 47

Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
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”
Total combined
Adminstration Total aerobic count
yeasts/molds count
route (cfu/g or cfu/mL)
(cfu/g or cfu/mL)
Nonaqueous oral 103 102
Aqueous oral 102 10
Most topicals 102 10
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Microbiological criteria :
 Environmental specifications: EU GMP, Annex – 1,
“Recommended limits for microbiological monitoring of clean
areas during operation”
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
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
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
× swab area
SESA

An example:

1000 cfu/g × 0.1 × 5 kg × 103

× 25 cm2 = 3943 cfu/swab
3170 cm2
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Quality Assurance
CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
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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Quality Assurance
 CLEANING VALIDATION…………...ACCEPTANCE CRITERIA
Determining acceptance criteria with more than one next
products (The Matrix approach):
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

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

Microbial swab (sterile) Chemical swabs (Texwipe) Cotton wipes

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Quality Assurance
CLEANING VALIDATION…………......SAMPLING METHODS
 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.
5 cm 2.5
(9) “Difficult to clean” equipment surfaces cm
shall be identified and sampled. 5 cm

(10) Representative surfaces of different Swab area

templates
materials (MOCs) should be sampled. 10
cm 55

Quality Assurance
CLEANING VALIDATION…………......SAMPLING METHODS
 Swab sampling techniques:
(11)Wiping should be unidirectional at a time. Parallel strokes
should be employed to cover entire swab area.

Courtesy: Validated Cleaning Technologies for Pharmaceutical Manufacturing, D. A. LeBlanc

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Quality Assurance
 CLEANING VALIDATION…………......SAMPLING METHODS
 Swab sampling techniques:
Example of “Difficult to clean” locations of an RMG:
Courtesy: Rapid mixer granulator, Kevin.

The design aspect of the equipment should be considered to

identify “difficult to clean” locations.
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Quality Assurance
CLEANING VALIDATION…………......SAMPLING METHODS
 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.
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Quality Assurance
CLEANING VALIDATION…………......SAMPLING METHODS
 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. 59

Quality Assurance
CLEANING VALIDATION…………......SAMPLING METHODS
 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
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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
 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.

HPLC pH meter

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Quality Assurance
 CLEANING VALIDATION…………ANALYTICAL METHODS

Specific Test Methods Non-Specific Test Methods

UV/Visible Spectrophotometry
Near Infrared Spectrophotometry
(NIR)
Total Organic Carbon (TOC)
High Performance Liquid
pH
Chromatography (HPLC)
Titration
Mid Infrared Spectrophotometry (MIR)
Conductivity
Atomic Absorption
Gravimetric
Capillary Zone Electrophoresis
Enzyme Linked Immunosorbant Assay
(ELISA)

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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
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
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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
 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……….
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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS

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

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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS

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

25 cm2
Revised swab area = × 5.5 μg / mL
4.0 μg / mL
= 35 cm2 (7 cm × 5 cm)
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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
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.

67

Quality Assurance
 CLEANING VALIDATION…………ANALYTICAL METHODS
Swab recovery schematic :
1. Spike control diluent directly

Control
Standard B μg/mL
solution
A μg/mL 2a. Spike
coupon

2b. Swab Control

coupon C μg/mL

2c. Extract swab

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Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
Recovery calculation 1 (Spiked against Standard):

(C μg/mL) × (mL)
% Recovery = × 100
(A μg/mL) × (mL)

 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.

69

Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
Recovery calculation 2 (Spiked against Positive control) :

(C μg/mL) × (mL)
% Recovery = × 100
(B μg/mL) × (mL)

 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.
70

Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
Rinse recovery schematic :

Case 1 Case 2
Pipette with
rinse solution
(known volume) Spike bottom
of SS beaker

Spiked
coupon
Lab sheker

Collection beaker 71

Quality Assurance
CLEANING VALIDATION…………ANALYTICAL METHODS
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.

72

Quality Assurance
CLEANING VALIDATION………………………HOLD TIMES

Cleaning Hold Time studies

Dirty Equipment Cleaned Equipment

Hold Time (DEHT) Hold Time (CEHT)

o DEHT = Max. allowed time, between end of usage and

employing cleaning
o CEHT = Max. allowed time, between end of cleaning and
further usage 73

Quality Assurance
CLEANING VALIDATION………………………HOLD TIMES
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

Quality Assurance
CLEANING VALIDATION………………………HOLD TIMES
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

Quality Assurance
CLEANING VALIDATION………………………HOLD TIMES
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

Quality Assurance
CLEANING VALIDATION………………………HOLD TIMES
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.
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Quality Assurance
CLEANING VALIDATION………………………HOLD TIMES
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.
78

Quality Assurance
CLEANING VALIDATION……………....ALL ASPECTS OF CV

Courtesy: Biopharm international

79

Quality Assurance
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
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
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
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
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
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
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
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
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
CLEANING VALIDATION…………USFDA 483 CITATIONS

Sampling Locations

“Exact / precise swab locations are not identified.”

89

Quality Assurance
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
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.”

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Quality Assurance
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
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
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
CLEANING VALIDATION…………USFDA 483 CITATIONS

Recovery Studies

“Equipment cleaning validation studies for…..did not

include…..present recovery studies on rinse
samples.”

95

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

Quality Assurance
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
100

Sambhujyoti Das, Quality Assurance

CLEANING VALIDATION