CONTAMINATION

CONTROL IN
CLEANROOMS

Shao-Min
Shao Min Yuan, Ph.D.
August 29, 2016

For those with allergies

Agenda

► What is cleaning validation?

► Cleaning
Cl i
► Cleaning activities
 Wiping

 Mopping

 Swabbing

► Sterilization & Disinfectants

► Swabbing & Validation

Cleaning Validation - Why

► US Food and Drug Administration (FDA) issued Guide to Inspections -

Validation of Cleaning Process in 1993

► Sources of Contamination of drug product or drug substance:

 can enter from the outside

 can be generated in the production environment during manufacturing

► Cross-contamination of drug
g either by:
y

 other active pharmaceutical ingredients (API) from previous batch runs

 by residues
es dues from
o the
e ccleaning
ea g age
agents
s used

► Can alter the strength, chemical identity of the drug

► Safety risk to patients consuming the drug

Regulatory Requirements

► 21 CFR- PART 211

 http://www.gpoaccess.gov/cfr/index.html

► § 211.67 Equipment cleaning and maintenance.

 (a)
( ) Equipment
q p and utensils shall be cleaned, maintained, and sanitized
and/or sterilized at appropriate intervals to prevent malfunctions or
contamination that would alter the safety, identity, strength, quality, or
purity of the drug product beyond the official or other established
requirements.
requirements

 (b) Written procedures shall be established and followed for cleaning

and maintenance of equipment, including utensils, used in the
manufacture,
f t processing,
i packing,
ki or h
holding
ldi off a d
drug product.
d t

Cleaning Validation - How

► Clean the equipment

 Develop
D l ththe method
th d ffor cleaning
l i

 Document this procedure

► Show that the equipment is clean

 Validation
• Develop procedure for recovering residue from process equipment
• Develop analytical method for sampling, detecting and quantifying residue
Goal of Cleaning Validation

► Confirm that the remaining residue in the production

equipment
q p of one product
p is below established acceptable
p
limits so that it does not impair the quality and safety of the
next product manufactured in the same equipment.

CLEANING

Where Clean Is Everything.™

Why Clean?

► Instrumental in a contamination control program

► Protect
P t t ththe product,
d t the
th process, and
d people
l
 Critical because the product is not processed further in its immediate container
and is vulnerable to contamination

 Airborne particles are significant because they can enter a product and
contaminate it physically, or, act as a vehicle for microorganisms and
contaminate it biologically

What to Clean and How Often?

ISO 5 Sample Cleaning Frequency Table

Surface Each shift Daily Weekly Monthly Quarterly
Trash X
Gowning room X
Floors X
Equipment X
Furniture X
D
Doors X
Windows X
Walls 2X
Ceiling X
Under raised floors X

The frequency is determined by the process needs!

Who Cleans?

► Operators (Associates, Technicians)

 Process
P – tools,
t l equipment
i t

 Worksurfaces

 Cleanroom

► Maintenance and Engineering staff

 Tools and equipment

 Worksurfaces and surrounding area

► Custodial staff

“Nobody wakes up in the morning excited about the prospect of cleaning surfaces at work”

Contamination

► What makes your

cleanroom dirty? Supply air

 Product and raw materials Adjacent

People
• Contamination is anything areas
showing where it does not
belong
 Process
• Lack of cleaning can result in Cleanroom
Product Materials
the process contributing to air
additional contamination of
your product
• Moving parts generate many
particles
Room Machines,
 People surfaces equipment

• We emit the most Water

contamination in a cleanroom source
Contaminants in the Cleanroom

Volatile
V l til Airborne
Organic Molecular
Contaminants Contaminants

Nonvolatile
Residue Microorganisms Particles
Adsorbed Ions
Molecules Metals
Fibers

Contaminants

► People generated microbial contamination

 Skin
Ski
• We shed 1,000 bacteria carrying skin particles per minute
• 1,000,000 bacteria can live on 1 cm2 skin

 Saliva
• Talking: projects droplets 2 – 3 feet
• Coughing: projects droplets 4 – 6 feet
• Sneezing: projects droplets 10 – 15 feet @ 200 mph
Good Practices

► Gowning
 Gown
G properly--
l garments
t fit,
fit snapped,
d tucked,
t k d masks,
k bouffant,
b ff t booties,
b ti etc.
t
• Check yourself in the mirror before entry
• Check co-workers
co workers

 Do not touch your face or goggles and do not blow your nose in a controlled
area – go into gowning room.

Sizes of Common Contaminants

Contaminant Size (µm)

Human hair 70 – 100
Human skin flakes 0.4 – 10
Pollen 5 – 100
Smoke 0 01 – 1
0.01
Household dust 0.05 – 100
Bacteria 0.25 – 10

► Depending on the person, the first visible size is ~40 µm

with the unaided eye
► The table shows that most of what you clean is not visible
Contaminant Generation Rate

Activity >0.3 µm particles per minute

M ti l
Motionless/Sitting/Standing
/Sitti /St di 100 000
100,000
Head, arm, neck, leg motion 500,000
Above adding foot motion 1 000 000
1,000,000
Standing to sitting or vice versa 2,500,000
g, 2 mph
Walking, p 5,000,000
, ,
Walking, 3.5 mph 7,500,000
Walking, 5 mph 10,000,000

► Limit as many fast motions as possible

 No running!

How is Cleanliness Maintained?

► Have procedures in place for the cleaning protocol

 Clean according to your company’s SOP only. Any changes may affect the
validation of the cleaning procedure.

► Be conscientious and mindful while cleaning

► Keep contamination to a minimum

 Work clean to allow easier clean-up later

► Have good cleaning practices for contamination

removal and control
FDA 483 Warning Letter Example

► Pharmaceutical manufacturer was cited for the following:

 “F
“Failure
il to
t clean
l andd maintain
i t i the
th packaging
k i equipment…
i t to
t preventt
contamination that would alter the safety, identity, strength, quality of the
products...

 FDA was concerned that the cleaning procedures and the line clearance
inspections were not able to detect that the affected equipment was missing
some of its parts.

FDA 483 Warning

g Letters are p
public information,, available on FDA’s website
http://http://www.fda.gov/ICECI/EnforcementActions/WarningLetters/2015/default.htm?Pag

How Particles Adhere to a Surface

► Van der Waals force

 Attractive
Att ti and d repulsive
l i fforces b
between
t molecules
l l
• Allows substances like hexane to form a solution

► Capillary
C ill adhesion
dh i force
f
 Attraction between unlike molecules
• Example: Water lifting in a straw

► Electrostatic forces
 Ionic interactions – opposite charges attract
• Example: Particles on TV screen, balloon on hair
How Particles Are Removed

► Wiping
 Physical action
 Use of a liquid decreases or removes the capillary adhesion force
 Surfactant – decreases capillary adhesion by reducing surface
tension
 Ionic surfactants modify surface and contaminant potentials and
reduce adhesion forces

Where are the tools used?

► Wipers
 Example surfaces to clean – small surfaces
• Worksurfaces
• Process equipment
• Associated equipment: carts and carousels
► Mops
 Example surfaces to clean – large surfaces
• Floors, walls, ceiling
► Swabs
 Example surfaces to clean – small surfaces
• Hard-to-reach (holes, slots)
• Irregular
► Frequency
q y
 According to process SOPs or WIs
Common Substrates

► Man-made fibers ►Natural fibers

 polyester – cotton
– cellulose
 nylon

 rayon

►Man-made
►Man made materials ►Blends
– foam
(combination of fibers)

– polypropylene – polyester/cellulose
►Composites
(sandwich of materials)
– polypropylene/cellulose
p yp py

Common Textures

►Knitted
►Hydroentangled
►Woven
►Composite
Man-Made: Continuous Filament Yarns

►Long, unbroken fibers

►Synthetic
– polyester
– nylon
►Range
► a ge o
of textures
e u es
– knitted
– hydroentangled
– woven
►Few particles and releasable
fibers

Man-Made: Continuous Filament Yarns

Knitted polyester Close-up of loop structure

Natural: Staple Yarns

►Short fibers
►Natural
– cotton
– cellulose
►Range of textures
– knitted
– hydroentangled
– woven
►Many particles and releasable
fibers
►Typically higher ion levels

Natural: Staple Yarns

Woven Nonwoven
Wiper Edge Treatments

►Particles and fibers are generated when

fabric is cut into wipers
►Three edge treatments are common
– cold-cut edge
– laser-cut edge
Laser-Cut Edge
– sealed border
►Most particles and fibers in the wiper
body are removed during laundering
►Overly rigorous washing can also
generate particles and fibers

Sealed Border

Substrate Summary

 Materials
– man-made
– natural fibers
– combinations
 Texture
– knitted
– woven
– nonwoven

 Edges
– cold-cut
– laser-cut
– sealed border
 WIPING

Where Clean Is Everything.™

Small Surface Cleaning and Disinfection - Wipers

Step 1 – Select the Proper Wiper

► Make sure your wipe is compatible with the cleaning or

disinfecting
g solution
 Cotton wipes are not compatible with quats

 Nylon wipes are not compatible with peroxides and quats

 Microdenier (100% polyester only) and polyester wipes work well with quats,
peroxides, bleach and IPA

► For sterile or aseptic areas, use sterile wipes and sterile

solutions

Step 2 – Preparing the Surface for Disinfection

► When using a one-step disinfectant, no pre-cleaning of the

surface is required
q before disinfectant application,
pp , if no visible
soils are present.
► Ap
pre-cleaning
g step
p is always
y required:
q
 When one-step disinfectants are not used (always before bleach, hydrogen
peroxide, some phenols and quats. Check the disinfectant label.)

 When the surface has visible soil, the soil must be removed before any
disinfectant use.

 When changing the type of disinfectant in a rotation

► Use different wipes for cleaning and disinfecting solutions

 IPA pre-wetted wipes are good for cleaning and removing residues
Step 3 – Spray the Wiper with the Solution

► Spraying the wipe, not the surface, is recommended

 If the
th di
disinfectant
i f t t solution
l ti isi sprayed
d di
directly
tl onto
t th
the surface,
f th
the surface
f
contaminants and solution droplets may be spread to adjacent areas

► Hold the nozzle close to the wipe for even coverage

coverage, but not
too far so as to spray the solution particles into the air
► Spray three to four times or until the wipe is at the appropriate
wetness level
 Damp
p for cleaning,
g wetter for disinfection

 Over-wet wiping is generally discouraged for cleaning

 Over-wet
Over wet wiping is required for disinfecting to leave surfaces wet for the
recommended contact time

Damp wiping

► Reduces capillary forces

► Solubilizes contaminants
from surface
► Transports contaminants
into the wiper capillaries as
a suspension in the cleaning
solvent
► Keeps dry contaminants
from being dispersed into
immediate environment
Over-wet wiping

► Negatives
 Redisperses contaminants
 Increases the chance of the
cleaning solvent acting as a
contamination source

► Positives
 Useful when soaking hardened
deposits
 Needed for disinfection

Step 4 – Wipe the Surface

► For the most efficient wiper use, quarter-fold the wiper by first
folding the wipe in half, and then in half again.

► Quarter-fold the wiper aids in applying a uniform pressure to the

surface
► Unfold
U f ld and
d refold
f ld th
the wiper
i tto expose a clean
l surface
f
Wiping Tips

► Wipe from a clean area to the

dirtiest, usually back to front (toward
you) on horizontal surfaces and from
top to bottom on vertical surfaces.

► Consistently
C i t tl move left-to-right
l ft t i ht or
1 2 3 4 right-to-left. Do not mix directions.

► Wiping should consist of parallel

parallel,
overlapping strokes. Each stroke
should overlap the previous stroke
by 20% for even coverage.
coverage Do not
use circular strokes or S-strokes as
these only spread contamination.

Step 5 – Disinfectant Tips

► Follow the disinfectant’s use instructions found on its label

 W
Wait
it the
th entire
ti exposure time.ti Th indicated
The i di t d exposure titime iis needed
d d tto
destroy the listed bacteria, viruses and fungi!

 If diluting a concentrate, be sure to follow the label use instructions and the site
cleaning and disinfecting protocols

► All disinfectants (phenol-, quat-, hydrogen peroxide plus peracetic

acid-, and bleach-based products) leave residues, which should be
removed according to site SOP
Step 6 – Remove Residues

► Use another wipe and residue removal agent.

► Spray the solution on the wipe or use a IPA pre-wetted

pre wetted wiper
► Repeat Step 4 wiping the surface to remove the residue

► In aseptic compounding areas, using sterile 70% IPA solutions is

required by USP<797> and using sterile wipers help maintain the
sterility of the IPA and the work area
► The residue removal agent could be 70% Isopropyl Alcohol (IPA) or
TexP (4% or 7.5% hydrogen peroxide solution)
► 70% IPA pre-wetted wipers are ideal for removing residues
 Higher IPA content improves drying rate
 Wetness level is optimized for efficient removal of residues

Wiping Technique Review

► Use parallel, overlapping strokes for effective cleaning and

disinfection
► Avoid introducing new contaminants while cleaning or
disinfecting
 Select
S l t the
th proper wiper
i substrate
b t t
 Watch chemical compatibility
 Control wiper migration (wipers approved for less clean areas ending up in
more clean areas)
► Do not over-saturate wiper with the cleaning solutions
► Change the wiping surface frequently to avoid re-deposition of
contamination
► Follow established cleanroom protocols
Wiping Guides

MOPPING

Where Clean Is Everything.™

Large Surface Cleaning and Disinfection - Mops

Step 1 and 2 – Select a Bucket System, Mop, and Disinfectant

► Choose a bucket system.

 Flat mops for walls and ceilings
 String or strip mops for floors.
► Confirm your mop material is
compatible with the disinfectant
class.
class
► Use sterile polyethylene bucket
liners (STX7099) to protect your
buckets from disinfectant
exposure and aid in easier clean
up.
► For sterile or aseptic areas, use
sterile
t il products
d t
 Wipers
 Disinfectants
 Equipment
Tips

► Ceilings and walls typically do not need a pre-cleaning step.

► Floors
Fl are typically
t i ll classified
l ifi d as “hi
“high
h ttraffic”
ffi ” and
d are lik
likely
l tto b
be
considered soiled, requiring a pre-cleaning step before disinfection.
► A pre-cleaning
l i ((rinse)
i ) step
t isi required
i d when
h changing
h i ththe ttype off
disinfectant in a rotation
► The pre-cleaning
pre cleaning step is not required when a one
one-step
step disinfectant
is used (for example, TexQ disinfectant), unless visible soil is
present.
► Use a cleaning solution for a pre-clean step. Some disinfectants like
TX6466 allow the use of the product for pre-cleaning, too, as
indicated on the product label.

Step 3 – Preparing the Disinfectant Solution in the Buckets

► Two-bucket system
 Make the disinfecting solution in Bucket 1.
 Fill Bucket 2 with the same solution (to avoid the dilution of the disinfectant in Bucket 1)
1).
Be sure to follow SOP.
► Three-bucket system
 Make the disinfecting solution in Bucket 11.
 Fill Bucket 2 with the same solution (to avoid the dilution of the disinfectant in
Bucket 1). Be sure to follow SOP.
 Leave Bucket 3 empty as it is used as the wring bucket to collect the dirty
solution. Refer to the disinfectant product label for correct solution preparation.
► Water should meet the quality of the water used in the
cleanroom
 Sterile water for sterile cleanroom or work area
► More is not better… Adding extra concentrate leaves more
residue that requires removal later
later.
Step 4 – Perform the Cleaning or Disinfectant Application Steps

A D B, E
► Single bucket system
Wringer
 Not recommended; however, it used

► Two bucket system (Life Science)

A Wet the mop with the disinfectant or
A.
cleaning solution from Bucket 1. 1 2
B. Wring the mop in Bucket 2.
C Mop
C. M th the surface.
f Disinfectant Disinfectant,
D. Rinse the mop in the Bucket 2. or water
E. Wring the mop in Bucket 2. (SOP)
F. Repeat Steps A through E until area is
completed. If, at any time, any solution Bucket 1 2
becomes cloudy or contaminated, the
solution
l ti mustt bbe replaced.
l d 2 Bucket 1

Step 4 – Perform the Cleaning or Disinfectant Application Steps

A D B,
B E
► Three bucket system Wringer
A. Wet the mop p with the disinfectant
solution in Bucket 1.
B. Wring the mop in empty Bucket
3.
1 2 3
C. Mop the surface.
D. Rinse the mop in the Bucket 2.
Disinfectant Disinfectant,
Disinfectant Empty
E Wring
E. W i th
the mop iin B
Bucket
k t33.
or water
F. Repeat Steps A through E until (SOP)
area is completed. If, at any time,
any solution
l ti bbecomes cloudy
l d or
contaminated, the solution must
be replaced. 3
2
Bucket 1
Tips

► The mopping solution (based on the two- or three-bucket methods)

must be changed
g after 600 square
q feet ((at a minimum)) for Class
100 and 1000 (ISO 5/6) and after 1,000 square feet (at a minimum)
for Class 10 000 and 100 000 (ISO 7/8). The mop head should be
re-wetted every eight linear feet
feet.
► Mop covers should be replaced according to your SOP, when visibly
dirty or no longer effectively applying or removing solution
solution.
► When disinfecting, the mop should be wet enough to leave a
solution layer on the surface to remain wet for the prescribed
contact time.
► When cleaning/using just a cleaning solution, the mop should be
well wrung out.

Mopping Technique – 1

Pull-lift method, like wiping

► For walls and ceilings
 Flat mop only
 Mop from a clean area to the dirtiest, usually
from top to bottom
1 2 3 4  Consistently move left-to-right or right-to-
left. Do not mix directions.
 Mopping should consist of parallel,
overlapping strokes. Each stroke should
overlap the previous stroke by 20% for even
coverage.
► For walls
 Do not touch ceiling or floor
► For ceilings
 Use four foot strokes, not over your head
 Do not contact or wet HEPA filters
Mopping Technique – 2 (Floors)

► Modified Figure 8 – AlphaMop or BetaMop

► Feels
F l more “natural”
“ t l”

About 4’ across

Start – from Bucket 1 Stroke 1

Stroke 2
Flip
Stroke 3

Finish – to Bucket 2 Stroke 4

Remember overlapping
strokes!

Step 5 – Disinfectant Tips

► Follow the disinfectant’s use instructions found on its label, which

include the contact time required
q to kill specific
p or described
microbes. If diluting a concentrate, be sure to follow the label use
instructions and the site cleaning and disinfecting protocols
► The indicated exposure time is needed to destroy the listed
bacteria, viruses and fungi
► All disinfectants (phenol-, quat-, hydrogen peroxide plus peracetic
acid-, and bleach-based products) leave residues, which should be
removed
Step 6 – Remove Residues

► Residues on floors need to be removed more frequently, perhaps

everyy day,
y, due to safetyy reasons (slipping
( pp g hazard),
), residue visibility
y
or possible product or process contamination.
► To remove residues, use the two- or three-bucket system
y with water
only, IPA solution (TX3290 sterile, TX117 nonsterile) or Hydrogen
peroxide solutions (TX684G and TX687G).
► Use the same water used in the room, e.g., sterile water for sterile
areas. Use the same cleaning steps and techniques as described in
Step 4
4.

Mopping Guide
Wiping and Mopping Protocol References

► “Standard Practice for Cleaning and Maintaining Controlled Areas and

Clean Rooms” ASTM E2042-04
► Cleanroom Housekeeping - Operating and Monitoring Procedures”, IEST-
RP-CC018.4
► D.
D Cooper
Cooper, “Basics
Basics of Effective Wiping
Wiping”, A2C2, Volume 4
4, Number 5
5, p
p. 47
47,
2001.
► L. Mainers, “How to Clean the Invisible”, Cleanrooms East 2000
Conference Proceedings .
► H. Siegerman, “Wiping Surfaces Clean”, Vicon Publishing, Amherst, NH,
2004.
► Anne Marie Dixon, “Cleaning of Non-Product Contact Surfaces” in
Cleaning and Cleaning Validation for the Pharmaceutical and Medical
Device Industries, Volume 1: Basics, Expectations
p and Principles”,
p Paul L.
Pluta, editor, DHI Publishing, River Grove, IL, 2009, pp 221 – 234.

STERILIZATION &
DISINFECTANTS

Where Clean Is Everything.™

Sterilized vs Sterile

滅菌 無菌
滅菌=無菌?

Why Disinfect?

► Prevent microbial contamination

 Protect the product being manufactured

 Minimize safety risk to patients consuming product

► Kill microbes that can cause infections

Types of Microbes

► Bacteria
 0.5-5 µm length
 Quickly replicate
► Viruses
 20-300 nm diameter
 Replicate inside living cells
► Fungi
 2-10 µm diameter
 Yeasts and molds
► Spores
 Most difficult to kill
 Require special claims - sporicide

Classes of Disinfectant

► Sanitizer
 Low
L performance
f disinfectant
di i f t t

 Not kill all bugs

► Disinfectant
 Higher performance in kill claims

► Sporicide (Sterilant)
 Highest
g p
performance in kill claims

 100% reduction of all microbes including spores

Kill Claims

► Determine what microbes are killed

► If do
d nott test,
t t dod nott know
k
 What microbes

 Contact time (how long to leave it on surface)

 Concentration of disinfectant

► Claims are based on directions of use

 Prescribed concentration

 Can be used “off label” but claims do not apply

FDA and Pharmaceuticals

► USP 34 General Chapter <1072> Disinfectants and Antiseptics

► USP <1072> is the most important information guide that the FDA would
point to for compliance

► FDA requires that end users must perform Disinfectant Qualification on

microbes found in their production areas

► Qualification = Efficacy Testing + Validation

► Improper, non-existent or inadequate procedures for disinfectant

qualification are one of the hottest topics with the FDA in 483 Warning
Letters

► Alcohol (IPA and Ethanol are NOT disinfectants)

Categories of Bugs to be Killed

Stronger
disinfectants

LESS powerful Source: USP<1072>

di i f t t
disinfectants

Qualification in user environment

 FDA enforces that end users must perform disinfectant efficacy studies on microbes
found in their production areas
 FDA requires end users to validate the use of the disinfectants in their local
production environment because their bugs may be different or unique, and
certainly more relevant and prevalent in their cleanroom

 EPA-registered kill claims provide end users with assurance that the selected
di i f t t will
disinfectant ill kill th
the prescribed
ib d b
bugs
 They also provide assurance that they may kill related or similar bugs found in their
environment,, compared
p to a product
p that has no EPA kill claims at all
 Kill claim testing and validation can be laborious and expensive, so end users typically
like to have the assurance that comes with EPA registered claims
Classes of Disinfectants

Sporicides

Bleach

Phenolics

Quaternary
Ammonium

Alcohol –
70% IPA

SWABBING & VALIDTION

Where Clean Is Everything.™

Swab uses

► Two major categories

 Cleaning, removing and applying substances
• Cracks and crevices
• Hard-to-reach locations
• Small
S ll surfaces
f (O
(Optics)
ti )
 Sampling – Cleaning Validation or Environmental Sampling (Sterile)
• Small locations on large surfaces
• Where cleaning is difficult
 Irregular surfaces
 Hard-to-reach areas
 Porous surfaces
 Heated surfaces
 Cracks and crevices

Cleanroom Swabs

► Sometimes Q-Tips just won't do

 Fabric
F b i options
ti ffor cleaning
l i validation
lid ti swabs
b

 Contamination characteristics of swab materials – selecting the optimum

material

 Cleaning Validation Fundamentals

 Sample the surface

surface, but don't
don t contaminate it in the process
Swab Materials

 Man-made fibers  Natural fibers

 Polyester  Cotton
 Nylon
N l  Cellulose
 Rayon

 Man-made materials  Blends (combination of fibers)

 Polyurethane foam  Polyester/cellulose
 Polypropylene  Composites (sandwich of materials
 Polypropylene/cellulose

Swab Functional Requirements

► Ability of the swab to recover the

residue from the surface
► Ability of the swab to release the residue
to an extracting solution for analysis
► Minimum contaminant release from the
swab to the surface
► Minimum
u blank
b a contribution
co t but o from
o the
t e
swab
Swab considerations

► Capture and Release residue of interest

► Particulate burden

► Chemical composition of the swab

► NVR (non-volatile residues)

Typical Swab Sampling Pattern

Swab 1 Side 1 Swab 1 Side 2

Step
p1 Step
p2

Flip swab
Fli b over
Change direction 90°

Swab 2 Side 1 Swab 2 Side 2

Step 3 Step 4 Step 5

Swab perimeter
using
i swabb titip
Flip swab over
Change direction 90°
Releasing Notched Swab Heads

► Rest notch on swab along top edge of vial

► Apply
A l pressure tto snap th
the swab
b
► Swab head falls into the vial

► Elute residues from swab head with solvent

► Make up solvent volume for analysis (TOC or HPLC)

► May need to filter or sonicate the solution before analysis

Analytical Methods

► Specific
 HPLC – UV-Vis,
UV Vi MS or CLND

 AA, ICP-MS, etc.

 LAL

 ELISA

► Non-Specific
 TOC

 pH

 Conductivity
Sampling – HPLC/UV-Vis or TOC

► Standard analytical techniques used for drug product residues - e.g.,

HPLC GC,
HPLC, GC etc.
t
► TOC often used for cleaning residues because of lack of organic
f
functional
ti l groups
► TOC not specific for particular substance

► Blank contribution from swab must be negligible - polyester is best

choice

How to Analyze the Sample

HPLC
HP C TOC

High Performance Liquid Chromatography Total Organic Carbon

Residue-specific Assay Non-specific Carbon Assay

A
Amount off each
h substance
b ppb
b ((parts per billion)
billi )

‘Picture of your sample’ Extremely Sensitive Assay

UV detection, typically, maybe MS Conductometric detection

HPLC

► Analytical Chemistry method, “High Performance Liquid

Chromatography”
g p y
► HPLC utilizes a column, a pump that moves the sample
g the column, and a detector that shows the retention
through
times of the molecules.
► Retention time varies depending on the molecules, compared
to a library

HPLC/UV-Vis

► Positives
 Good sensitivity to functional groups in drug actives
 Good specificity
 Good for APIs
 Technology well-established

► Negatives
 Can be expensive
 Prone
P to
t interferences
i t f
 Requires functional group for response – does not detect detergent residues
TOC

► Positives
 Simple to measure
 Minimal method development
 Relatively inexpensive
 Detects detergent residues which lack functional groups

► Negatives
 Non-specific – cannot provide characterization as to the type of residue detected
 Any organic carbon source is an interference
 Must have back-up selective method if validation fails

Swabbing Guide
Cleaning Validation Articles

"Applications of Total Organic Carbon Analysis to Cleaning Validation", Jenkins et

al PDA Journal of Pharmaceutical Science & Technology,
al., Technology VoVo. 50
50, No
No.1,
1 Jan
Jan- Feb
1996, p6.
 Used 2"x2" polyester wipers for sampling

"Total Organic Carbon Analysis of Swab Samples for the Cleaning Validation of
Bioprocess Fermentation Equipment", Strege (Lilly) et al., BioPharm, April 1996, p
42.
 Used wetted polyester swabs for sampling.
 Mean recoveryy of 82% for E. coli cells for samples
p spiked
p at 64 µg
µg/swab.

Searching for Nothing

Cleaning Validation:
“I have found nothing and that’s good news.”
http://www.texwipe.com/files/pdfs/published_papers/search_for_nothing.pdf
Swab is not just a swab

Cleaning Validation:
Why the swab matters
http://www.texwipe.com/files/pdfs/published_papers/swabs_matter_in_cv.pdf

Quiz Questions

1. Why does one need to do cleaning validation?

A. Government regulatory bodies require a show of proof that the production area is clean.
B. To prevent cross contamination that may alter the strength and/or the chemical identity of the
drug.
drug
C. All of the above

2. Why does the US FDA not recognize 70% alcohol as a disinfectant?

A. 70% alcohol does not kill any microbes.
B. 70% alcohol evaporates too fast to perform efficacy tests.
C. 70% alcohol does not leave any chemical residues after drying

3. What is the correct sequence/direction for cleaning?

A. Dirty to clean
B. Clean to dirty
C. Direction is not important as long as you wipe/mop/swab in overlapping strokes

4. What are the three main selection criteria when choosing a wiper?
A. Material, Texture, Edge treatment
B. Material, Size, Texture
C. Texture, Edge treatment, Size

5. Why is TOC a popular analytical method for cleaning validation?

A.
A Because it measures the amount of CO2 in the environment
B. Because the government requires one to perform the TOC test
C. Because TOC is most effective in identifying cleaning detergent and disinfectant residues.