Sampling for

Cleaning Validation

Dawn Tavalsky
Outline
•Types of sampling
–Direct sampling
–Indirect sampling

•Case Studies in deviations dealing with sampling

What Types of
Sampling are we
Talking About for
Cleaning Validation?
 Cleaning Validation Sampling

Direct Sampling
To measure actual members of a population
in cleaning validation – to measure the actual
equipment being cleaning (e.g., to take a swab sample
of the equipment surface)
Indirect Sampling
To measure records or surrogates related to the actual
members of a population
In cleaning validation – to take a related measure rather
than sample the actual equipment (e.g., to look at the
contaminants in the rinse water)
Sampling the Baby or the Bathwater
Even though the “baby” is in the bathtub
– clean bathwater doesn’t necessarily
always indicate that the baby is clean – so
we typically sample both the baby (direct
sampling) and the bathwater (indirect
sampling)
Let’s Look at Direct Sampling First
 Types of Direct Sampling

Visual Inspection (qualitative)

 To physically look at and examine the direct surface of the
equipment to confirm that contaminants, impurities, residual
product, etc are removed
 Visual Inspection employs some or all of the following:
Naked Eye (prequalification eye exam may be required)
Boroscope
Mirrors
Extension poles
Bright lighting
White cloths
Black cloths
 Coupon studies are often preformed to quality the level to which a
soil can be seen
 Types of Direct Sampling

Swab Sampling (quantitative)

 To physically wipe the surface of the equipment using a swab to
confirm that contaminants, impurities, residual product,
bioburden, etc are removed
 Swab sampling employs some or all of the following:
Swab(s)
Solvent (the liquid material the swab is dipped into prior to
sampling)
Diluent (the liquid material the swab is placed into after
sampling)
Extension poles
Sample bottles
 Coupon studies are preformed to quality the level to which a soil
can be recovered utilizing the chosen swab and swab solvent
Distance Impact on Viewing
Detection and Light Intensity
Sometimes the Nose Knows
Sometimes You Just Feel
 Swab Samples

A swab sample is prepared by physically swabbing a specified area of

a piece of equipment with a swab with the intent to extract any
collected residue from that swab for analysis.

• Ideal for residues not easily removed with water rinsing

• Swabs can physically remove insoluble residues
• Chosen swabs must exhibit three important characteristics:
– Ability to recover desired residue from given surface
– Ability to release the residue into an extraction solvent
– Must not contribute excessive interference or
background during analysis
 Swab Samples

Specific Swab (supplier and part number)

Swab Solvent vs. Sample Diluent
Number of swabs
Dry vs. Wet Swabs
Desired Surface Area (Ex: 10x10 cm or 4x4 in)
Swabbing Patterns
 Swabbing Techniques
Examples of different swabbing techniques are shown below.
Various techniques should be evaluated to determine which will
provide the highest residue recovery.

• Continuous Strokes
(Zig-Zag)

Flip Swab
• Discrete Strokes

Flip Swab
 Swabbing Techniques

• Non-Flat surfaces

Flip Swab
Flip Swab
Let’s Look at Indirect Sampling Next
 Types of Indirect Sampling

Rinse Sampling (quantitative)

 To physically collect rinse water that has been passed over the
equipment in question. The absence of contaminants, impurities,
residual product, etc in the rinse water infers the absence on the
equipment surface
 Rinse sampling employs some or all of the following:
Qualified water source, often Water For Injection
Sample bottles
Catch basins (when sampling small parts)
 Coupon studies are sometimes preformed to quality the level to
which a soil can be recovered utilizing the chosen rinse water type
and temperature
Cleaning Validation Sampling
•Direct Surface Sampling
• We typically perform surface sampling for analytical samples such as
TOC and product specific markers. May also surface sample for
bioburden

•Indirect Rinse Sampling

• We typically perform rinse sampling for analytical samples such as TOC,
LAL, Bioburden, Conductivity, pH, etc
Let’s Take a Minute to Look at The
Tools Used for Sampling
Let’s Take a Minute to Look at The
Tools Used for Sampling

http://www.globepharma.com/html/remote_swabbing.html
 Let’s Take a Minute to Look at The
Tools Used for Sampling

http://www.ge-mcs.com/en/remote-visual-inspection/video-borescopes.html
Let’s Take a Minute to Look at The
Tools Used for Sampling
Let’s Take a Minute to Look at The
Tools Used for Sampling
Let’s Take a Minute to Look at The
Tools Used for Sampling

Charles River
Lonza
Let’s Take a Minute to Look at The
Tools Used for Sampling

GE Sievers
Shimadzu
Teledynetechmar
Let’s Take a Minute to Look at The
Tools Used for Sampling

Mettler Toledo
MANY Others
Let’s Take a Minute to Look at The
Tools Used for Sampling
 Which Types of Sampling
Should I use During My Cleaning Validation?
• Start with visual detection
• If visual detection alone proves insufficient (in most cases –
visual detection alone is not accepted as sufficient) – move to a
Quantitative analysis
• Quantitative methods provide greater specificity, accuracy and
sensitivity
• The sampling techniques selected must be capable of
quantitatively determining the amount of residual material on
the manufacturing equipment.
• The two most commonly used sampling techniques to choose
from are swab (directly sampling the surface) or rinse samples.
 Swab or Rinse Sampling?

Considerations:
Ease of access for sampling
Worst case locations
Equipment disassembly
Ability to sample safely
Size of equipment
Solubility characteristics of compound of interest
Physical removal
Chemical removal
 Swab Samples

Advantages:
“Direct” surface sampling
Focus on worst case locations
Physical removal of residues

Issues:
Access to sampling locations
Manual procedure – Training Personnel
Choosing appropriate swab
 Rinse Sampling

A rinse sample is a collected aliquot of the solvent used to

perform an equipment rinse to remove any remaining residue.
Applicable for:
•small surface areas
•difficult to reach areas (cannot disassemble equipment)
Solvent should reach all surface areas
Surfaces should be rinsed long enough to ensure complete
coverage and sufficient removal of the target residue
More simplistic than swabbing procedures
Sample is generally collected from final rinse of equipment
 Rinse Sampling

Advantages:
• Sample inaccessible locations
• May use different solution from process rinse
• Provides overall picture

Issues:
• Solubility of residue in rinse solution
• Need to relate amount in rinse sample to potential
contamination of next product
Deviations Related To Cleaning
Validation Sampling
Case #1 Cleaning Monitoring
Failure

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Case #2 Environmental Monitoring
Failure
Case #2 Insufficient Drain Line
Case #3 Rouge
Case #4 Water Spotting
Case #5 Heat
Scorching
Gravimetric Last to Leave Determination
Coupon Dissolution
Determine Hardest To Clean Soil
from each soil Group

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Gravimetric Last to Leave Determination

Coupon Preparation
Soil Preparation
Dissolution
Evaluation
Worst Case Soil Determination

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 Gravimetric Last to Leave Determination

Coupon Preparation
Select MOC that matches your process
Properly Label Coupons
Thoroughly Clean Coupons Prior to Analysis
Drying, Desiccation, and Storage of Coupons
Tare Weight

48
 Gravimetric Last to Leave Determination

Coupon Selection
MOC
In-Process Soils
Temperatures
Detergents
Size/weight
DOE: Analysis Desired Teflon 316SS
#8
316SS
#8
Viton Noryl

We Used 316SS #8 2”x2” Poly- 316SS H202 Valox Glass

propylene #7 Cured
Silicone

Aluminum 316SS Titanium Pt-Cured Buna-N

#4 Silicone Rubber

49
 Gravimetric Last to Leave Determination

Labeling
Should Identify MOC
Should Reference
Certificate of Manufacturing
Should have a unique
Identifier

We use Marking Methods Inc. Electro-Chemical Marking Equipment

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 Gravimetric Last to Leave Determination
Cleaning: Clean with a method and detergent compatible with the MOC
Coupon for Current Gravimetric Dissolution Studies are being:
Sonicated for 20min in 3%CP310
Rinsed 3X with Milli-Q H20
Dried under a Biosafety Hood
Visually inspected
Smudges are removed with Alcohol
Stains are passivated with 5% H2SO4
Dried at 125C for >20 minutes
Desiccated for >30 minutes

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Gravimetric Last to Leave Determination

Coupon Cleaning
Sonication in 3% CP310

Rinsed with Milli-Q H20

Dried under a Biosafety Hood

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 Gravimetric Last to Leave Determination

Coupon Cleaning
Drying

Desiccation

Storage

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 Gravimetric Last to Leave Determination

Weighing
Tare Weight of the MOC Coupon is the
Most Important Measurement
Make sure balance is level
Wear gloves or handle coupons with
forcepts
Make sure Coupon is dry and free from
debris
Accurately record coupon ID
Wait for mass measurement to stabilize
before recording weight
Record out as far as possible

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 Gravimetric Last to Leave Determination

Soil Preparation
Accurately measured (Volume, Mass,
Dose)
Reproducible Application
Pipette
Dry/Desiccate/Store
Proper Handling
Document Visual Observations

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 Gravimetric Last to Leave Determination

Dissolution (standardization)

Volume and Concentration of Detergent

Temperature

Turbulence

Time

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 Gravimetric Last to Leave Determination

Distek Model 2100C Water Bath

Dissolution Unit
200RPM
500ml Detergent
50°C
Time (+~30sec)

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 Gravimetric Last to Leave Determination

Distek 2100C Controls

Hydraulic Lift (Time)
Set Temperatures
Set RPMs
Set Paddle Height

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 Gravimetric Last to Leave Determination

Evaluation of Data
Dry Soil Weight Pre-Cleaning
Dry Soil Weight Post Cleaning
Percent Soil Loss

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Questions?
More Information?

Dawn.tavalsky@genzyme.com
Feel Free to Contact me
with Questions.

Thank you for your

attention