This document would not be possible

without the generous support of the

following BPSA sponsors:
Published by:
Bio-Process Systems Alliance
BPSA c/o SOCMA
1850 M Street, NW
Suite 700
Washington, DC 20036
www.bpsalliance.org

AUTHORS
Ken Davis, Value Plastics, Inc.

Patrick Evrard, GSK Vaccines

Eric Isberg, Pall Life Sciences

Ernie Jenness, EMD Millipore

Mike Johnson, Entegris

Csilla Kollar, Dow Corning Corporation

Mark A. Petrich, Merck & Co., Inc.

Helene Pora, Pall Life Sciences

John Stover, AdvantaPure/New Age Industries

Kirsten Strahlendorf, Sanofi Pasteur

James Dean Vogel, The BioProcess Institute

Maureen Eustis, The BioProcess Institute

1
Table of Contents
Topic Page
Part I: Introduction 3
Part II: Particle Risk 4
Part III: Particle Characteristics & Quantification 6
Part IV: Particle Measurement Methods 9
Part V: Single-Use Technology Lifecycle 11
Part VI: Methods of Control for Suppliers 14
Part VII: Particulate Evaluation as Part of End User Manufacturing 17
Part VIII: Deviation Response/Mitigation Plans 18
Part IX: Summary and Conclusion 20
Part X: BPSA-recommended Next Steps 22
Part XI: Terms and Definitions 23
Part XII: References 25

Figures
Figure 1: Risk from particle contamination by category 5
Figure 2: Potential particle locations 6
Figure 3: Particle sources in SUT 7
Figure 4: Particle size 8
Figure 5: TAPPI size estimation chart 9
Figure 6: Fishbone of potential particle sources 11
Figure 7: Sources of possible particle contamination 11
Figure 8: Summary of sources of particles – BASE CASE 12
Figure 9: Summary of sources of particles – SUPPLIER improvements to particulate contributions 13
Figure 10: Summary of sources of particles – with END USER filtration step added 13
Figure 11: ISO 7 cleanroom particle counts by week 16
Figure 12: Use of controls over time to reduce particulates 21
Figure 13: Example of end user/supplier agreement for particulate acceptance criteria 22

2
Recommendations for Testing, Evaluation, and Control
of Particulates from Single-Use Process Equipment
Part I: Introduction
The Bio-Process Systems Alliance (BPSA) is an industry association whose mission is to facilitate, globally, the
development and manufacturing of biopharmaceuticals through the implementation of robust, safe and sustainable
Single-Use Technologies. Corporate members include plastic equipment suppliers, service providers and users in
the biopharmaceutical industry who share this mission. A key focus of BPSA’s core activities is to educate its
members and others through sharing of information and development of best practice guides that help suppliers,
users and regulators to safeguard the quality of drugs produced with single-use process technologies.

Intent and Scope
This document provides recommendations to suppliers and
end users in the single-use technology (SUT) industry
regarding particulate testing, evaluation, and control. The
practices presented here are intended to help characterize
levels and types of particles, as well as to provide methods
to assure minimal levels of particulate in SUT.
strive for fewer particles and fewer types of particles in the
SUT.
In this document, BPSA members provide a view to help the
SUT community navigate this complex topic throughout the
lifecycle of the SUT.
This document helps clarify:

Background  Why are particulates an issue with SUT?

The Bio-Process Systems Alliance (BPSA) recognized a need
in the SUT industry and started discussions about
particulates at its 2012 and 2013 International Single-Use
Summits. Attendees showed significant interest, and a
working group comprised of subject matter experts (SMEs)
from SUT supplier and end user companies was formed to
discuss the topic and recommend best practices. This
document captures those discussions with more specific
content and formal references in order to guide the SUT
community.
BPSA members recognize that achieving zero particles is not
physically possible for process equipment used in filling of
injectable drug products, and the goal is to get as close to no
particles as possible. We add single-use technology suppliers
to the goal stated by Stephen Langille in his paper entitled
Particulate Matter in Injectable Drug Products:
 We clarify why particles in SUT may be a contamination
risk to:
o the active ingredient and/or formulated product;
o the biological cells which produce the product; and
o the patient who is treated with the product.
 How does one control particulates during the manufacture
of SUT?
 How does one control particulates during the use of SUT?
 How does one address situations when particulates are
found in or attributed to SUT?
This paper's intended audience is a combination of those
who are new to SUT as well as those who are already
familiar with the technologies. It is anticipated that the
document will provide a base for both groups to gain a
common understanding of this complicated topic.

“The goal of end users, regulators, and standards-
setting organizations should be to minimize
particulates in drug products, without placing
unnecessary expectations on suppliers for minimal
safety gains. Improving the manufacturing quality will
reduce the risk of harm to patients from particle
contamination.”1
In summary, for SUT fluid paths, fewer particles/particulate
is better, and whereas zero is the goal, it's not an achievable
specification. This means the SUT industry must continue to
This document is also intended to help both the end user
and the supplier. It can help the end user convey their
specific requirements to the supplier. In turn, suppliers can
use the document to demonstrate what they are capable of
providing to the end user.
Definitions of terms are provided at the end of this
document to assist in the ongoing dialog among end users,
suppliers, and regulators.

3
Part II: Particle Risk
A particle is loose mobile matter or embedded matter
that is unintentionally present in/on the single-use
component/assembly and potentially may contact or may
end up in the process/product fluid.
When it comes to particulates in biopharmaceutical
processing, fewer is better! Both SUT suppliers and end
users are taking steps to decrease the levels of unwanted
particulates in their products. This is in order to minimize
the risk of contamination of the drug products, and
consequently, decrease the risk of harm to patients from
particulate contamination.
Reducing and controlling particles in SUT is important,
especially for those that are used for final formulation and
filling downstream of final filtration, or for aseptic
manufacturing of vaccines and therapies that cannot be
filtered.
Background
It is critical to understand the types and sources of
particulates and how their presence may affect the use of
SUT. Regulatory guidance very clearly spells out the
expectations for equipment, their surfaces, and any
particulate that may be contained within the equipment.
In the US FDA Drug cGMP, CFR 211.65 states:
“Equipment shall be constructed so that surfaces that
contact components, in-process materials, or drug
products shall not be reactive, additive, or absorptive
so as to alter the safety, identity, strength, quality, or
purity of the drug product beyond the official or other
established requirements.”
US FDA cGMP for Biologicals, CFR 600.11 similarly states:
“All surfaces that come in contact with products shall
be clean and free of surface solids, leachable
contaminants, and other materials that will hasten the
deterioration of the product or otherwise render it less
suitable for the intended use…”
Comparable statements can be found in regulations or
guidance from all regulatory authorities, relating to
injectable API (active pharmaceutical ingredient), bulk drug
substance and final drug or biological product containers.
The presence of particulates can potentially affect the
patient, as well as alter the release of drug substance, drug
product, and intermediates:
Patient – The clinical significance of particulate matter and
how foreign debris from medical products and devices can
cause or exacerbate patient complications is well
documented. The particles can cause mechanical obstruction
of micro blood vessels, leading to intravascular coagulation,
thrombosis, or distal embolization. The immune system can
be overtaxed by particles, and particles can affect the
efficacy of the drug (AAMI TIR42:2010).
Cell growth – Particulate matter can interfere with cell
growth through either viable (bioburden) particle
contamination or non-viable particle interference.
Drug product – Product quality can be affected by the
presence of particles. Particulates have affected drug
products by affecting their safety, purity, effectiveness, their
basic chemical structure and/or their bioavailability to the
patient. In 2012, there were over 37 particulate-related
recalls of drug product.2
Intermediate – The production process may also be
affected by particles. Similarly, process intermediate purity,
chemical structure, and reactivity may be influenced by the
presence of particles.
Particle Risk
Risk is usually defined as the probability of harm multiplied
by the severity of harm. Good pharmaceutical quality
represents an acceptably low risk that the product will fail to
achieve the desired clinical attributes.3
Risk knowledge helps to focus improvement efforts on the
most critical aspects of the process. Risk awareness is
important during process development, technology transfer,
monitoring and control. 4
SUTs are often employed to provide closed-system
processing with ready-to-use sterilized components and
assemblies. Traditional pharmaceutical multi-use equipment
is usually cleaned before sterilization. During these cleaning
processes, the surfaces are rinsed with cleaning chemicals
and high purity water. The objective of these cleaning
processes is to remove soil and environmental
contaminants. This includes reducing the levels of surface
particulates.
During manufacturing, SUTs usually do not undergo a final
rinsing step. Particulates that may be generated during their
manufacture or come from the manufacturing environment
may remain on the SUT’s fluid contact surfaces.
The characteristics and the potential risks presented by the
SUT can be viewed in terms of where particles are found,
and the types of particles present in the SUT.
4
Major classifications for SUT particle risk are:  Viable (bioburden) or non-viable;
 Size of particle: Visible (typically greater than 100
Process application (the application or use of the SUT): microns) or sub-visible;
 Storage;  Appearance of the particle: Shape, texture, color (usually
 Transfer; able to be captured by photography, optical microscopy or
 Mixing; scanning electron microscopy (SEM).
 Upstream; Chemical make-up of the particle:
 Filling; and  Characterization to the elemental level can be carried out
 Cell therapy. using a variety of different analytical techniques, including
Location of particle in or on the SUT: SEM/energy dispersive X-ray spectroscopy (EDS), micro-
 Product contact surface (usually the interior of the SUT); raman spectroscopy, micro-FTIR (Fourier transform
 Exterior of the SUT; and infrared spectroscopy) and PLM (polarized light
 Embedded in the SUT material. microscopy).
Particle characteristics: Count of the particles:
 Intrinsic or extrinsic;  Extent of particle contamination.

Figure 1 below summarizes the categories of particle risk and illustrates the risk continuum that should be assessed for each
specific SUT application.

Figure 1: Risk from particle contamination by category

LOW HIGH
RISK
Process Application Media Final Fill

LOW HIGH
Packaging RISK Process
Location of Particles Contact
Exterior Embedded Surface

LOW HIGH
RISK
Size of Particle Small Visible
Large

LOW HIGH
Chemical Make-up Native RISK
Toxic
of Particle (Material of Construction) USP Class VI

LOW HIGH
RISK
Quantity of Particle None Many

Process Application usually characterized by short exposure times, and the

downstream particle risk is reduced significantly by
The importance of the presence of particles varies with the
filtering. This is not always possible, and results are
application of the single-use technology. In general, the
dependent upon the appropriate choice of filter medium and
closer the process is to the patient, the greater the risk.
form.
Each process application has its own particle risk potential.
SUT handling practices should be assessed carefully. The
Time of exposure and the type of operation (e.g. storage,
impact of the presence of particles is significantly greater for
transfer, or mixing) also directly affect the potential risk of
processes that cannot be filtered (e.g. final fill, cell therapies,
an application.
and some vaccines). SUTs used for these applications
Many process steps employ filtration, resulting in the without pre-use rinsing need to meet higher particulate
removal of particulates. These filtration processes are quality requirements and lower overall particle content.

5
Location of Particles
Particles may be present on the surfaces of, or embedded
within, the materials of SUT components and assemblies.
This is illustrated in Figure 2 below.
Figure 2: Potential particle locations
Exterior
Process/Product
SUT Surface
Contact Surface
(interior)
embedded within
component
Process/product contact surface: Loose particles present
on the wetted contact surface form the most common
particulate exposure and the majority of particulate risk to
patients receiving injectable drugs. These particles are in
direct contact with drug products and usually on the interior
of the SUT. Some SUTs are inside other SUTs and their
exterior surface is also involved in process/product contact,
e.g., an impeller inside a bag.
Process/product contact particles, their reduction and
addressing their risks, form the primary focus of this
document.
Exterior SUT surface: Particles present on the outside of
the SUT are usually a low risk to most processes. Most SUT
processes are closed and routinely not exposed to the
exterior of the SUT. Exterior particles, however, may affect
other adjacent processes, cleanroom or isolator operations
and should be addressed appropriately.
Embedded: Embedded particles in SUT components and
assemblies may create a risk to the process depending on
their location and whether they are fully encapsulated or
exposed to the process contact surface. Embedded particles
have the potential to affect product performance.
Gels may be present in plastic film and sheet. According to
ASTM D883 Standard Terminology Relating to Plastics, gel in
the film or sheet is to be distinguished from contamination
such as particles of dirt, carbon, or lint.
Risk Summary
To summarize risk:
The goal of suppliers, end users, regulators, and
standards-setting organizations should be to minimize
particulates in drug products, without placing
unnecessary expectations for minimal safety/quality
gains.
Ultimately, this will reduce the risk to patients from
contaminating particles.
Part III: Particle Characteristics &
Quantification
Particle Definition
Particulates can be defined as small “pieces” or “parts” and
are synonymous with particles. Various formal definitions
have been provided in the literature (AAMI, ASME, etc.). In
the biopharmaceutical industry, where typical drug matrices
are aqueous-based, detectable particulates are solids or
immiscible liquids. The USP <788> Particulate Matter in
Injections definition of particulate matter says:
“Extraneous mobile particles, other than gas bubbles,
unintentionally present in solutions.”
The BPSA definition includes SUT-specific language:
A particle is loose mobile matter or embedded matter
that is unintentionally present in/on the single-use
component/assembly and potentially may contact or
may end up in the process/product fluid.
Type of Particle
Particles found in SUT are classified by their source and the
ability to identify them by the following categories:
Intrinsic (native) particles are created from the process
contact materials of the SUT or the drug process. Intrinsic
particles are part of the process/product formulation
ingredients, packaging materials, etc. They are reasonably
expected to appear and sometimes are generated with the
use of the SUT. These particles are all non-viable and do not
contain living organisms. Endogenous particulates are a type
of intrinsic particle and are derived from constituents of the
pharmaceutical process or product.
 Extrinsic (foreign) particles are not related to process
contact materials. Extrinsic particles typically include
rubber, metal, plastic, hair, fibers, insect parts and dust
coming from the manufacturing environment. They can be
introduced at various steps of the SUT manufacturing
process and the drug manufacturing process.
6
 Known – These can be identified and are usually tracked
to their source with trending along with intrinsic particles
and listed in the fishbone diagrams in Part V: Single-Use
Technology Lifecycle (page 4).
 Unknown – THESE ARE OF MOST CONCERN. They can
be difficult or impossible to identify. Their presence casts
doubts about quality systems and environmental controls,
because unidentified sources may not be resolvable.
Figure 3: Particle sources in SUT
INTRINSIC
(SUT)
EXTRINSIC
(Known & Unknown Sources)
Particle Characteristics
As they all pose different risks to patients, the key particle
characteristics to consider when evaluating particulates are:
 Particle size
o Visible or sub-visible
 Particle appearance
o Shape
 Spherical or angular
o Hardness
 Soft/deformable or hard/brittle
o Texture
 Smooth or rough
 Particle chemical composition.
Particle Size
Particles can be classified by their size, specifically whether
they are visible or not visible to the naked eye.
Depending on the size, there are various pharmacopeial
monographs in existence that provide standards for particle
testing in finished sterile dosage forms. In the absence of
specific guidance for SUT, these pharmacopeial references
have been applied to rinse effluents from SUT.
 Viable particulates (bioburden) also belong to the
extrinsic particle category and their viability can be
mitigated with gamma irradiation microbial control or
sterilization, provided the validated bioburden levels are
maintained within the controlled levels. Residuals from
the elevated bioburden may be toxic, e.g. endotoxin.
See Figure 3 below.
Methods Environment
Materials People
UNKNOWN
The USP <1> Injections “essentially free of visible particles”
concept for injectable drugs suggests that zero particulates
is the desired goal. Similar descriptions exist under the
European and Japanese Pharmacopeias using slightly
different language: “practically free” of particulate (EP
2.9.20) and “free from readily detectable” particles (JP 6.06)
with unaided eyes. There are two additional USP
monographs under development to address visible
particulates. USP <790> Visible Particulates in Injections will
be setting allowable limits for visible particles instead of the
“essentially free” particulates standard set by USP <1>. USP
<1790> Particulate Matter Determination: Visual Inspection
will offer methods for visual inspections.
The draft USP monographs have clearly defined visible
particles as being >100 microns, approximately the size of a
grain of sand. The BPSA agrees with this definition and
views it as the lower limit for visible particles. The shape
should also be considered when assessing the size of a
particle because they are not always a perfect sphere.
Therefore, detection methods may not accurately detect
asymmetrical particles, e.g. a fiber that is longer than 100
microns, but has a diameter of much less than 100 microns.
The BPSA also suggests the 100-micron size be the
benchmark for differentiating between visible and sub-
visible particles.
7
Figure 4 below shows the range of particle sizes.
Figure 4: Particle size
10 microns 25 microns
0.13 microns (USP <788>) (USP <788>)
0.3 microns
influenza
virus red
blood
animal
brev cell
cell
diminuta
bacterium
Particle Viability
Vegetative microbial cells or spores capable of germination
and generating live cells may be considered viable particles.
These are usually introduced to the SUT from the
environment and personnel. Even low levels of gamma
irradiation can provide a significant level of microbial
control and sterilization ensures absence of viable cells
(viable particles), but their presence and potential
byproducts, e.g. endotoxin, however, could still contribute
risk to the patient or pharmaceutical process from the SUT.
Particle Appearance
The appearance of the particle is important to characterize it
and identify its source. Observing and recording shape,
color, and texture help differentiate the types of particles.
Digital photographs with magnification or a microscope also
assist in documenting the particulates.
Particle Chemistry
The chemical make-up also assists in identifying the
particulate and its potential risk. Different materials
contribute particles in different ways. Materials of
construction of the SUT, and the end user’s other production
system parts, are often the source of particulates in SUT. In
addition, cleanroom supplies and clothing are common
sources of particles discovered in SUT.
100 microns
(USP monographs)
sub-visible
visible
grain of sand
The possible types of particulates are those from the SUT,
the machines and tools used to make them, and the
environments where they are made. These can include:
 Intrinsic (SUT materials, such as components, tubing, bags,
etc.):
o Dark-colored particles that may be embedded.
 Extrinsic:
o Known source:
 Environment (pollen, bacteria, etc.);
 People (skin flake, hair, etc.);
 Methods (manufacturing equipment, use, handling,
etc.);
 Materials (fibers, metals, ceramics, glass, paper,
adhesives, polymers, etc.) – gowning, machines,
facility, cellulosic (paper, packaging materials).
o Unknown source – AREA OF MOST CONCERN.
Analytical techniques including SEM, EDX, Micro-Raman
spectroscopy, and Micro-FTIR, are employed during
investigations to identify particle compositions.
Particle Count
Particle count is the number of particles per unit area or
particles per unit volume of the SUT. This indicates the level
of contamination. Particle counts are recorded by qualified
methods and trended.
8
Part IV: Particle Measurement
Methods
Particulate contaminant assessment methods must be
repeatable and capable of representing the actual level of
particulate in the SUT. Each measurement method must be
fully qualified and administered by properly trained
personnel with qualified instruments.
Photo by AdvantaPure/New Age Industries
Particulate measurement methods have evolved from
mostly non-intrusive observation methods, e.g. visual with
no magnification, to vision systems with magnification, to
more and more intrusive methods that analyze liquid
samples.
Visual Inspection
Visual inspection has been carried out by both the suppliers
and the end users. It primarily verifies the proper product
configuration and the presence of observable visual particles
(>100 microns).

Typical attributes inspected for are:

 Component arrangements are verified;

 Connections are verified to be secure;
 Embedded particles/gels;
 Film creases;
 Exterior particles are observed and removed;
 Finished goods are inspected for internal particles where
possible, e.g. clear and translucent components; and
 Actions are taken based on inspections and may include
discarding the single-use assembly.

Inspection using a light table

Visual inspections can be enhanced with magnification, light tables, and particle size comparison charts, e.g. TAPPI chart (see
Figure 5 below).

Figure 5: TAPPI size estimation chart

100 microns=0.1 mm; 20,000 microns2=0.02 mm2

Reprinted with permission of TAPPI

9
Visual inspection can also be limited by the product
attributes and by human and method capabilities. The visual
method is less effective for components that are opaque or
have intricate internals. Visual inspection may not be able to
assess accurately the entire level of particulate
contamination, especially through one or more layers of
packaging.
Liquid Measurement Methods
Liquid measuring methods for free particles in SUT have
evolved from those methods used for final formulation, e.g.
USP <788>, because of the absence of a standard or another
methodology for more appropriate assessment of the level
of particles within the SUT. This is one area that may benefit
from industry standardization to ensure alignment and
proper assessment of particulate levels within and among
SUT assemblies.
Ultimately, a specific particulate study of the SUT applied to
the process, or a representative "worst case" model system,
is the best way to measure the amount of particle risk from
that SUT. These are destructive tests, and the SUT cannot be
used after testing. These tests are typically performed
during qualification and/or routine monitoring of the SUT
manufacturing process, not as a pre/post-use test.
The USP <788> Particulate Matter in Injections and its EP, JP
and WHO equivalents (JP 6.07, EP 2.9.19 and WHO IP (5.7))
set limits for sub-visible particulates. Informational chapters
such as USP <1788> Methods for Determination of
Particulate Matter in Injections and Ophthalmic Solutions
provide procedures to count “extraneous, mobile un-
dissolved particles other than gas bubbles, unintentionally
present in the solutions” 10-25 and over 25 microns in size.
These monographs were written with the express intention
of preventing contamination in “solutions for injection
parenteral routes” and they describe the process for testing
of materials in filled drug vials and acceptable limits for
particulates found by those methods. Such testing has
recently been extrapolated to packaging and SUT. Although
size cutoffs at 10 and 25 μm are stipulated by USP <788>,
there is increasing industry interest in smaller particles.
There are other guidelines besides the pharmacopeias that
have been developed to study particulates released from
device surfaces. AAMI TIR42 references USP <788> for
counting particulates, but it does not address particulates
that arise from degradation or wear and does not consider
liquids as particles. This report recommends additional size
ranges to be considered for evaluation. The ASTM F24
Standard addresses counting particles over 5, 25 and 100
microns found on the surface, or washed from the surface
using sonication and detergent. It also discusses two
magnifications (45x and 100x) for counting particulates on,
or washed from, the device surface.
Before counting, the collection of the particulates from the
contact surface for analysis is very important. Homogenous
liquid drug solutions are measured to determine compliance
with USP <788>. In order to evaluate SUT particulates, liquid
must be used to rinse the product contact surface of the SUT
in some manner.
The count will be affected by many different parameters,
and these should be addressed during the study design. The
parameters include dimensions of the SUT, handling of the
systems before study, temperature, pressure, harshness of
the particle removal, and effluent age before analysis,
amongst others.
The study conditions will affect the success and
reproducibility of capture and counting. When studying
particulates, it is vital to understand and control the capture,
count, and characterization of particulates.
Sample preparation and particulate extraction with particle-
free water should follow an approved standard operating
procedure (SOP) to ensure repeatability of the sample
collection. Wetting of the full fluid path, agitation to loosen
particles and extraction liquid collection are all variables
that need to be consistent from test to test. The total rinse
volume should be minimized while still achieving the
mobilization, suspension, and extraction of the particle
content of the test article.
USP <788> describes two techniques for enumerating
particles in or from filled dosage containers; light
obscuration and membrane microscopy. Once the sample is
collected, particle testing is generally performed using
Method 1 of USP <788>, particle counting using a light
obscuration particle counter. Method 2, the microscopic
method, is also employed at times, where more visible and
sub-visible particles are captured, observed, and counted,
and as a reference to differentiate gas bubbles or liquid
droplets like silicone oil that can influence light obscuration
measurements. This method also allows for cataloguing of
the observed particles, but is also labor-intensive and
subject to human error.
USP <787> Sub-visible Particulate Matter in Therapeutic
Protein Injections and USP <1787> Measurements of Sub-
visible Particulate Matter in Therapeutic Protein Injections
will be published in the near future and will additionally
address particulates less than 10 microns. It will also outline
multiple analytical procedures for the measurement and
characterization of particulates like dynamic imaging,
Coulter analyzer, turbidimeter, light diffraction, and light
scattering.
The techniques with the potential to be used eventually in a
quality-controlled environment are expected to be flow
imaging and counting based on the Coulter principle (apart
from light obscuration).5
10
Part V: Single-Use Technology A common approach of Particulate Management Programs
utilizes a fishbone of five potential sources for each step of
Lifecycle the SUT lifecycle to address the particulate risks in a root
Ultimately, the level of particles needs to be suitable for the cause method (see Figure 6 below).
intended use. Particle levels present need to be anticipated
so all parties involved can assess the risk that the SUT brings Figure 6: Fishbone of potential particle sources
to the process.

Ideally, the SUT would be assembled in a “clean build” ma me pe

manner with the objective of continuous reduction of the ch
ine th op
levels of particles present. The objective in this document is od le
to help begin to define the requirements that SUT end users
have and allow SUT suppliers to show how they can meet or
s
strive to meet those requirements.
me n ts n t
u re on m e
meas
Each step of the SUT’s lifecycle can contribute to the
potential for additional particulates. A proper Particulate
Management Program will minimize these contributions and
envir
keep the levels as low as possible. Process steps can be
added, e.g. filtration, to reduce SUT’s particulate levels.

Each component of the SUT assembly brings its own set of potential particulate risks, and the numbers can grow very quickly
as demonstrated in Figure 7, below.

Figure 7: Sources of possible particle contamination

ma me pe
ch ine tho op
ds le ma
ch me pe
ine tho op
ents nt FILTER ds le
urem onme
meas envir ts
a s u remen onme
nt
me e n vir
TUBING
CONNECTORS
ma me pe
ch ine tho op
ds le
SAMPLE
ents nt
a s urem onme
me envir ma me pe
ch ine tho op
ds le
ents nt
urem onme PRODUCT BAG
meas envir

ma me pe
ch ine tho op
ds le
ents nt
a s urem onme
me envir

11
Each step in the SUT lifecycle also contributes potential The particle levels should be tracked and trended in order
particulate risks (see Figure 8, Figure 9 and Figure 10 on to monitor performance and measure the effect of
pages 12 and 13). This increases the potential risks of improvements or process upsets. When applied correctly,
particles even further: these trends are like instruments on a plane’s dashboard,
indicating where the particle levels are, and where they
Supplier: are most likely headed. The pilot (supplier or end user)
 Raw materials of the components (resins, compounds); now has information to react to and make appropriate
 Preparation of equipment (cleaning and/or sanitization) corrections for better management of the levels of
for manufacturing components or assemblies; particles.
 Fabrication of each individual component. (bag film,
tubing, connectors, O-ring, filter membrane); Two phases in the lifecycle of an SUT need to be considered
 Assembly of the sub-assembly (bag with ports, filter and integrated when analyzing particle generation and
capsule, connector); control. The first phase includes the supplier’s sources,
 Assembly of the final assembly (bag assembly, transfer manufacturing and assembly, packaging and shipping. The
assembly, filling assembly); second phase includes end user receiving, handling, and use
 Packaging; of the SUT. Particles are introduced to, and generated within,
 Transportation and handling; and the SUT in both phases.
 Sterilization (irradiation).
The list above shows some of the ways that end users can
End user introduce particles to SUT.
 Receipt;
 Quality inspection;
 Storage;
 Transfer to the production area;
 Removal of packaging (may occur at various stages);
 Preparation (e.g., rinsing, autoclaving);
 Use; and
 Disposal.

Figure 8 (see below) is a depiction of a summary of sources of particles in a single-use system. The left side illustrates the
possible particulate contribution from the supplier during manufacturing (machine, methods, people, materials, and
environment). The right side shows the end user particulate contribution prior to or during use.

Figure 8: Summary of sources of particles – BASE CASE

SUPPLIER CONTRIBUTION END-USER CONTRIBUTION

USE
RAW SUB-
MATERIAL
PARTICLES
+ COMPONENT
PARTICLES + ASSEMBLY
PARTICLES
+ ASSEMBLY
PARTICLES = END-USER

+
(e.g. Pump)
=
includes
sum of total
number of

RAW MATERIALS COMPONENTS SUB-ASSEMBLY ASSEMBLY

particles
-
(e.g. Filtration) END-USER

Thermoplastics connector

Thermoelastomers tubing
sample device
Adhesives sample device

Metal filter

Glass bag

Ceramics

TIME
PARTS SUB-ASSEMBLY PACKAGE, STERILIZE, SUPPLY CHAIN TRANSFER, UNPACKAGE, PREP, USE, FILTER

12
Figure 9 (see below) is a depiction of how supplier best practices are applied to reduce levels of particles in the SUT assembly,
and how they can reduce the levels of particulates from the product baseline, and thus reduce the overall particulate risk.
Fewer branches of the fishbone diagrams on the supplier’s side lead to less risk. This is shown by smaller circles than those in
Figure 8 (see page 12).

Figure 9: Summary of sources of particles – SUPPLIER improvements to particulate contributions

SUPPLIER CONTRIBUTION END-USER CONTRIBUTION

USE
RAW SUB-
MATERIAL
PARTICLES
+ COMPONENT
PARTICLES + ASSEMBLY
PARTICLES
+ ASSEMBLY
PARTICLES = END-USER

+
(e.g. Pump)
=
includes
Fewer branches adding particles decreases sum of total
number of
particle count throughout process
particles
-
(e.g. Filtration)

Figure 10 (see below) is a depiction of how the end user can reduce the presence of larger particulates by utilizing a filter in
their process, and thus reducing the overall particulate risk. This leads to smaller circles than those shown in Figure 8 and
Figure 9.

Figure 10: Summary of sources of particles – with END USER filtration step added

SUPPLIER CONTRIBUTION END-USER CONTRIBUTION

USE
RAW SUB-
MATERIAL
PARTICLES
+ COMPONENT
PARTICLES + ASSEMBLY
PARTICLES
+ ASSEMBLY
PARTICLES = END-USER

The
+
addition
(e.g. Pump)
=
of a filter can decrease the number
includes
sum of total of large particles contributing to the end-user’s
number of final product.
RAW MATERIALS COMPONENTS SUB-ASSEMBLY ASSEMBLY
particles
-
(e.g. Filtration)
END-USER

Thermoplastics connector

Thermoelastomers tubing FILTER

sample
sample
Adhesives device
device
Metal filter

Glass bag

Ceramics

TIME
PARTS SUB-ASSEMBLY PACKAGE, STERILIZE, SUPPLY CHAIN TRANSFER, UNPACKAGE, PREP, USE, FILTER

13
Chain of Responsibility: Assembler-
Manufacturer-Manufacturer’s Suppliers
While particulate control is the responsibility of each stage
of the supply chain, it is ultimately the final assembly
provider who needs to minimize particulates most. The final
assembly provider is responsible for ensuring that the
assembly meets requirements and contains minimal
particulate.
In addition to controlling its own environment and
implementing procedures to eliminate particulate in its
operation, the final assembly provider needs to understand
the particulate risks that can come from each component,
and hold each component provider to the same or better
level of particulate controls used by the final assembly
provider.
Each component provider should understand where
particulate can come from in the operation and what type of
particulate this is likely to be. It is a good idea to have quality
agreements in place that define acceptance criteria, as well
as suggesting inspection procedures to be put in place to
identify and remove particulate before component
packaging.
Each level of the supply chain needs to assess any packaging
materials involved with their component and the packaging
of each component.
As with the components, this packaging should contain a
minimal level of particulate. It should also be determined
whether components will need to be multi-bagged,
especially if they are going to end up in a cleanroom for
assembly.
Part VI: Methods of Control for
Suppliers
Manufacturing of single-use assemblies typically occurs
within a cleanroom, an ISO 8, Grade D, Class 100,000 or an
ISO 7, Grade C, Class 10,000 environment. Although this
space is very clean, it is not certified as a “particle free”
space. Therefore, additional controls should be utilized to
ensure an assembly meets the acceptable particle criteria,
enabling filled injectable products to meet pharmacopeial
standards e.g., USP <1> for visible particulate and USP
<788> for sub-visible particulate. The basic requirements to
minimize particulates during the manufacturing of single-
use assemblies are as follows:
 Manufacturing processes designed to minimize the risk of
particle generation, introduction or inclusion into the
finished assembly;
 Maintaining proper preventative maintenance of
manufacturing equipment;
 Ensuring cleanliness of materials and people entering the
cleanroom;
 Controlling flow of materials and personnel within the
manufacturing environment;
 Providing operator training;
 Demanding cleanroom gowning;
 Ensuring cleanroom maintenance and control;
 Inspecting product, including up to 100% in-process
surveillance and lot release testing;
 Recording performance trending for both the cleanroom
operation and the manufactured SUT; and
 Documenting non-conformance, root cause analysis and
corrective/preventative actions.
Raw Components
Monitoring and control of incoming materials, such as
tubing, fittings, bag film, filters, connectors, tie wraps and
packaging components is critical to ensure proper
particulate management. Simply put, the lower the particle
load of incoming materials, the higher the probability that
single-use assemblies with minimal particles, both visible
and sub-visible will be manufactured and shipped.
An initial component qualification for cleanliness of all
materials used in the manufacturing of SUT is
recommended. In addition to visual inspection, evaluation of
particle load in the flow path is recommended using an
appropriate and qualified method for particle identification
and bioburden evaluation, such as AAMI, ANSI, ISO 11137,
etc. Alignment with the component supplier is vital.
Discussions should be conducted with suppliers to ensure
they understand the criticality of their parts in the
manufacturer of the SUT.
Assuring that the component suppliers are vigilant in their
practices is an absolute requirement. The purchase
specification of components should state the requirements.
The component supplier should have systems in place to
ensure conformance to the particle requirements. Audit of
the component supplier is also recommended to ensure
controls are in place and proper handling and packaging
methods are utilized to ensure low particle and bioburden
load.
An incoming inspection procedure for components may be
required at the SUT manufacturer to ensure material is not
damaged during shipping and that it conforms to the
requirements listed in the purchase specification document.
Cleanroom Operation
A standard operating procedure should be in place for the
flow of materials into the cleanroom, and for final
assemblies leaving the cleanroom.
It is recommended that incoming materials be multiple-
bagged. This allows for the discarding of the outer bag, a
source of contamination of cellulose fibers and other
materials generated during shipping. The inner bag(s) may
then be sanitized with alcohol or other disinfectant or
cleaner and passed into the clean material storage area.
14
 nonviable) in the cleanroom is highly recommended to
verify performance of the room to the cleanroom
specifications. An established method for periodic
monitoring of the room under dynamic conditions is
paramount to ensure contamination levels remain
significantly below the accepted levels per the cleanroom
classification.

The monitoring process should encompass the entire facility

and take into account how areas relate to each other
throughout the monitoring period. Additionally, to ensure
Photo by PortaFab
proper operation of the cleanroom, monitoring of
temperature and humidity levels, as well as air differential
Example of a gowning area pressure between each classified area and the non-classified
area, is performed.
Inspection and cleaning for particulates on raw materials
generally occurs during the kitting of materials prior to
moving these materials into the clean assembly area. During
the assembly process, work surfaces should be cleaned on a Photo by EMD Millipore
regular basis. Operators should be inspecting for particulate
at the assembly operation. The final assembly is inspected,
multiple-bagged in the cleanroom and transferred to the
packaging area outside the cleanroom for final packaging.

In addition to ensuring clean materials, procedures and

training should ensure a high level of operator cleanliness. A
detailed procedure for an operator’s introduction into the
cleanroom is necessary to limit the transfer of particulate
from outside the cleanroom to within it. A dedicated
gowning room at a minimum of ISO 8 is recommended.

This room is set up with clean and dirty barriers to minimize

contamination transfer. The SOP should outline the proper
conduct associated with entrance, operator conduct, and exit
from the cleanroom. Procedures for gowning and de-
gowning should be clearly written, enforceable, and easily
trained to, in order to ensure maximum cleanliness of the
operator. The SOP must outline the “do’s and don’ts” for
cleanroom operation and include a description of articles
which are allowed in the cleanroom and those that are A gowned operator applying sanitizer to gloves prior to
restricted from the cleanroom. Recommendations can entering an ISO 7 cleanroom
include restriction of certain street clothing and/or
An annual recertification for compliance to ISO 14644-1 is
cosmetics worn by the operator prior to gowning.
required. The HEPA filters must be tested for airflow and
Reassignment to non-product contact activities is
pressure drop and aerosol particle challenge. This ensures
recommended in the case of illness.
that the HEPA filters and HVAC system are functioning per
Only approved cleanroom materials are utilized by the design to ensure proper removal of contaminants and the
operators including lint-free cloths, shoe covers, head appropriate number of air changes in the room per hour.
bonnets, beard covers, and powder-free gloves. Training to
Cleanrooms should be cleaned on a periodic basis to assure
the cleanroom SOP must be regularly performed. Operators
proper operation and particle control. Routine operation
are encouraged to “self-police” each other in order operate
and its impact on particulate levels should be evaluated and
at the highest level of cleanroom practice.
cleaning frequencies should be adjusted accordingly to
assure cleanroom operation. Additional cleaning usually
Cleanroom Performance occurs in response to non-routine events: planned
Demonstrated process capability of the cleanroom's (maintenance) and unplanned (deviations). Such events
environmental monitoring is another important aspect of should be evaluated for their potential impact on particulate
SUT manufacturing. Periodic (e.g. daily, weekly, monthly, levels and additional cleaning and/or rigor of cleaning
quarterly) monitoring for particulates (viable and should be adjusted to suit the nature of each event.

15
Figure 11 (below) indicates the particle counts for an active cleanroom. Of importance is the separation between the
maximum particle counts measured, the alert level and the maximum particle counts (max) allowed. Action levels are typically
set at 50% of the maximum particles allowed. The alert level is typically 25% of the maximum level.

Figure 11: ISO 7 cleanroom particle counts by week

Figure 9: ISO 7 Clean Room Particle Counts by Week

Particulate Levels

ACTION
W/ DEVIATION
SEASONAL ALERT and a
EXCURSIONS CAPA
ACTION
LIMIT

ALERT
LIMIT RETURN TO
NORMAL
PROCESS
CAPABILITIES

TREND
7/6/2009 1/22/2010 8/10/2010 2/26/2011 9/14/2011 4/1/2012 10/18/2012 5/6/2013 11/22/2013 6/10/2014 12/27/2014

Manufacture of the SUT
Manufacturing equipment and processes used to make the
SUT play an important role in minimizing particles in the
clean manufacturing space. Equipment should be
manufactured with materials that are cleanroom
compatible. This equipment should not shed particles or
fibers, be easy to clean, and designed in a manner to
minimize areas where particles can build up.
Proper filtration of process air, and venting of pneumatic
valves utilized in automated equipment, should be a design
consideration and then properly maintained.
Equipment placement should be done to avoid obstruction
of environmental airflow (intake and return) and include
proper preventative maintenance and operational controls
to minimize the risk of particle generation or accumulation.
Visual inspection is recommended before, during, and after
all process steps. These include the following:
 Port bonding;
 Film sealing;
 Final assembly of tubing, injection-molded fittings, bags,
filters and connectors;
 Leak testing; and
 Packaging.
Operators and management should be vigilant about particle
detection and improvement opportunities. The visual
inspection criteria should be clearly established, with all
operators trained and regular visual acuity testing
implemented, to ensure qualified inspection. Use of proper
lighting is important, e.g. light box/table. Final product
evaluation to agreed-to acceptance criteria is recommended.
This should be monitored, and its variation over time should
be measured to assess trends and demonstrate repeatability.
Testing per qualified methods (e.g. visual inspection and
USP <788>) is suggested.

 Bag manufacturing;
 Film cutting;

16
Although USP <788> is qualified as a sub-visible particle
test, the counts of particle greater than 25 microns alert the
manufacturer to potential visible particle issues.
If the assembly fails the release test, material should be
quarantined, and an investigation launched to assess the
reasons for the out-of-specification test result. Corrective
actions may be put in place after the investigation to limit
failures in the future.
Part VII: Particulate Evaluation as
Part of End User Manufacturing
Two phases in the lifecycle of an SUT need to be considered
and integrated when analyzing particle generation and
control. The first phase includes the supplier’s sources,
manufacturing and assembly, packaging and shipping. The
second phase includes end user receiving, handling, and use
of the SUT. Particles are introduced to, and generated within,
the SUT in both phases.

Table 1 (see below) lists some of the ways that end users can introduce particles to the SUT.

Table 1: Potential sources of particulates contributed by end user single-use operations

Source type Manufacturing-induced source

Processing materials and raw Particulates from the single-use component can interact with components of a protein
material ingredients/ product solution to form precipitates.6 These can be further exacerbated by process conditions
and/or type of single-use component

Manufacturing activities Connecting and disconnecting assemblies

Manufacturing environment
Using fiber-shedding filters with zero-to-minimal flushing
Limited use of rinsing/washing/flushing steps
Valve use
Pump use
Onsite or site-to-site transportation conditions and containers
Mismatched components, non-optimal component-equipment integration
Mixing components chafing inside of container or impeller parts/bearings
Rough handling
Regular equipment/processing aid wear
Abrasive product (e.g. undissolved aluminum salts)
Open system applications of single-use

Personnel Handling of SUT assembly or part(s)

Processing Considerations Photo by Sanofi Pasteur

Evaluations of particle risk in end user manufacturing

should include placebo runs that use worst-case process
conditions. Worst-case conditions are those that are
reasonably expected to occur and that present the greatest
potential for particle generation.

Parameters to consider are:

 Mixing speed;
 Number of connections made during the process;
 Storage times and temperatures;
 Line clamping or valve use;
A formulator in a development lab handling a
 Pumping/spallation;
single-use system carefully
 Rinsing/flushing/washing steps; and
 General handling practices

It is recommended to run the process in the intended

production facility or, at minimum, under the process and
product manufacturing conditions that are planned.

17
Best Practices for Handling Single-Use
Components
Although all of the items below may not directly affect
particles, these are recommended practices for SUT in
general:
1. Cover sharp parts. Do not remove supplier’s protective
coverings until necessary.
2. During storage, bags should be contained in a hard-
shelled container or, at minimum, covered with a sealed
outer bag. Lines should be secured as appropriate,
especially when freezing.
3. Flush the systems, especially those that contain filters or
fiber-shedding components, where possible.
4. Avoid over-processing: over-mixing, or over-handling of
components/assembly.
5. Avoid pulling, flattening, rubbing, squeezing, flexing, or
twisting of components/assembly.
6. Optimize the welding and sealing conditions to avoid
“flashing” or inadequate welds.
7. Keep product fluid contact path as short and with as few
components as possible.
8. Do not lift items by their tubing connections.
9. Minimize the stress on tubing junctions. Avoid sharp
bend radii.
10. Do not allow sharp objects to be used in the same area
as single-use components.
11. Match peristaltic pump tubing type and dimensions to
pump heads, process duration, and process fluids. Do
not exceed anticipated tubing life.
12. Minimize surfaces that can rub together during
shipping, storage, or use.
Part VIII: Deviation Response/
Mitigation Plans
When a visible particle is observed in an SUT, whether
during manufacturing or at the end user’s facility, the
response depends on a number of factors. Perhaps the most
important consideration is whether or not the particles are a
deviation from the quality standard or specification.
Purchasing specifications and inspection criteria should
make the particle requirements clear. Number, size, and
type of any allowed particles on or in an SUT should be
agreed to between supplier and end user via a clearly
written specification. The specification should take into
consideration the process capabilities of the supplier as well
as the quality requirements of the end user. It is important
to identify gaps between supplier capabilities and end user
expectations.
The next sections follow a single-use item through its
lifecycle and point out the actions that might reasonably be
taken to investigate visible particulate observations. The
factors to consider in making product disposition decisions
are also described.
A good particle investigation requires strong cooperation
between end users and suppliers. Both parties need to avoid
defensive or accusatory attitudes that do not facilitate the
open communication and clear thinking needed to resolve
problems quickly.
When in the SUT Lifecycle is the Particle
Observed?
The stage in the SUT lifecycle affects the type and intensity
of investigation activities. Consider the following when
preparing an investigation plan:
Particle found during SUT subcomponent
manufacturing and assembly:
 Investigate as a cleanroom management deviation.
Particle found during QC inspection prior to packaging
by the supplier:
 More extensive investigation is required; characterize the
particle and establish a root cause; document for statistical
process control (SPC) purposes; evaluate “related lots”
(those in the facility at the same or adjacent processing
times and lots prepared with common components).
Particle found after packaging and before shipment to
end user:
 More extensive investigation is required including a check
on the QC processes and their effectiveness. Particle
characterization is needed and a root cause should be
pursued aggressively. Document this for SPC purposes.
Particle found at end user receiving:
 This is potentially not a quality system problem for the
end user. A commercial complaint will be filed, provided
that the part in question is out of specification. An
investigation should be performed by the supplier for a
post-packaging event. Shipping configuration and method
should be assessed as potential contributing root causes.
Particle found at point-of-use (before use):
 This is a quality system problem for the end user and
needs to be assessed for impact on the cleanroom
environment. Root cause investigation needs to consider
both end user and supplier as potential sources of the
particles. A commercial complaint will be filed to obtain
supplier assistance and the event counts against supplier’s
quality performance score provided that the supplied part
is out of specification. An investigation should be
performed by the supplier as for a post-packaging event.
Particle found at point-of-use (during use):
 This is a quality system problem and quick decisions are
needed about continuation of processing and quarantines
of the end user’s product and SUT. Root cause
determination needs to consider both end user and
supplier as potential sources of the particles. A
18
 commercial complaint will be filed to obtain supplier
assistance. An investigation should be performed by the
supplier as for a post-packaging event
Particle found after processing is complete:
 Actions taken are the same as for those at point-of-use.
End user decision-making about quarantines will be
urgent.
Where is the Particle Observed – On or In the
SUT?
Particles that contact the end user’s process/product stream
are significantly more serious than those on the exterior of
the SUT or packaging materials. While process/product
contact surface (wetted path) cleanliness has top priority,
the mode of use of SUT may warrant concern about particles
outside the wetted path.
For example, an SUT used in non-closed system fashion or
within a Grade A environment needs to be as close to
particle-free as possible. Unusual requirements like this
need to be discussed with the supplier during the design
process.
A general approach for reacting to particles based on their
observed location is as follows:
Particles observed on the outer package (bag):
 These are usually not handled as an end user quality
system problem.
Particles observed on the inner package (bag that was
over-packed in a clean environment):
 This is a quality system problem for both the supplier and
end user and requires a root cause investigation. A
commercial complaint should be filed if this is out of
specification. The SUT will likely not be used. Potential for
similar particles to be in other units in the lot or in similar
SUT from the same supplier facility should be assessed.
Particles observed on the exterior of SUT or embedded
particles:
 The investigation intensity depends on the nature of the
particles, SUT specifications, and the stage of end user
processing as described above. The supplier should be
notified and a complaint filed as appropriate so that the
observations can be included in trending analyses.
Particles observed within the process/product contact
surface (wetted path) or in areas that connect with the
wetted path such as vent lines and unused tubing legs:
 This is a serious quality system problem as discussed
above. If the SUT is being used, quick decisions are needed
about continuation of processing and quarantines of end
user product and SUT. Root cause determination needs to
consider both the end user and supplier as potential
sources of the particles. A commercial complaint will be
filed to obtain supplier assistance. An investigation should
be performed by the supplier as for a post-packaging
event if it is found to be outside of specification.
Particle Investigation Steps
Given the general considerations about when and where a
visible particle is observed, the following steps are typically
taken for investigations involving suppliers, end users, or
both. Some of the steps are unique to end users and their
relationships with patients and regulatory agencies. Risk
posed by particles must first be considered in terms of
patient risk. Roles and responsibilities are dependent on the
specific situation.
Step 1. Detect a particle (or particles) during SUT
manufacturing, quality inspection, or during end user
operations.
Step 2. Report the finding via formal quality system
procedures (deviation report, event notification, etc.).
Suppliers and end users should alert each other as required
by quality agreements or other agreements.
Reporting should occur even if the SUT is within
specification so that trending analyses can be performed.
Step 3. Hold the lot (if possible) or continue at risk.
Immediate discard may be required (see below). The end
user’s product batch is placed under administrative
quarantine (and possibly physical quarantine).
Step 4. Quarantine related SUT parts as a precaution until a
root cause investigation provides more information.
Step 5. Capture and characterize the particle(s). Count if
there are too many to characterize each one.
Step 6. Compare the particles to a catalog of previously
found particles.
Step 7. Use characterization data and catalog matches to
determine the source of the particles and a root cause for
their appearance in the SUT. Materials of construction of the
SUT, and the end user’s other production system parts, are
often the source of particulates in SUT. Cleanroom supplies
and clothing are also common sources of particles
discovered in SUT. Look for obvious sources before chasing
exotic hypotheses. Analytical data is often tenuous for small
particles. Partner with the analytical services provider to
understand both what the compositional assignment is as
well as what plausible alternate assignments are. Knowledge
of SUT materials provided by the supplier and the end user
19
component engineer will help the analyst to run appropriate Part IX: Summary and Conclusion
tests and assist in the interpretation. Information about
suppliers who are located earlier in the supply chain may Improving the quality of SUT manufacturing, in order to
also be needed to identify the source of particles. generate the least amounts of particles, will simultaneously
satisfy the goals of the end users, suppliers, and regulators,
Step 8. Assess the impact on other SUT parts. Add while ultimately protecting the safety of the patient.
quarantines and, if necessary, remove quarantines or make
rejection/discard decisions. There are four primary areas that must be managed in order
to ensure robust control of particulates in single-use
Step 9. Assess the end user product batch impact and make
systems:
“disposition” recommendations. Product quality decisions
depend heavily on where the particles are found, the ability
of the investigation team to prove that similar particles are 1. Cleanliness of the incoming materials;
not present in other locations, and on the chemical identity 2. Cleanliness of the manufacturing steps and assembly
of the particles. Positive quality decisions (those that allow processes;
further use or release of a batch) are not possible without a 3. Cleanliness of the operators and associated gowning;
convincing root cause investigation. In the absence of and
sufficient detail, end user product should be discarded. 4. Cleanroom facility and equipment maintenance and
controls.
Step 10. Assess the impact on previously
manufactured end user products and notify regulatory
authorities if required by end user policy and/or law. Along with visual inspection and testing for particle levels in
Step 11. Prepare an investigation report and obtain the product, attention to these four areas will ensure the end
approvals for root cause analysis and product disposition user of minimum visible particulates in the SUT product.
decisions.
Step 12. Review investigation and quality decisions
with regulatory body representatives during inspections as
required. Consider this activity when preparing the
investigation report and supporting documentation.
Step 13. Identify and complete corrective and
preventative actions that may involve the disposition of the
batch, disposition of single-use parts and further steps to
prevent repeat investigations. Actions may include changes
to either the end user’s manufacturing process and/or
changes to single-use supplier manufacturing, packaging, or
sourcing.

20
Figure 12 (see below) illustrates how effective controls can reduce the number of particulates, specifically those from outside
sources. Less particulate is better!
Figure 12: Use of controls over time to reduce particulates
Environment
Environment
People
controls People
SUT SUT
Components Components
“Initial” “Final”
Particulate Profile Particulate Profile
Recommendations
Control of particles in SUT manufacturing requires
diligence from all suppliers in the supply chain, as well as
with the assembly of the system. Particle control is
everyone’s responsibility, including the end user. Because
manufacturing and use of systems are not performed in a
particle-free environment, systems should be implemented
to ensure cleanliness and monitor performance.
An ISO 8 or better environment is recommended for SUT
assembly and component manufacturing. Some assemblers
currently manufacture in ISO 7 and ISO 5 environments to
better control their manufacturing environments. In
addition, extra diligence, and steps such as pre-cleaning of
parts by the component supplier, may be required to
ensure parts meet the strict incoming needs of the SUT
manufacturers.
Particle control, both visible and non-visible, in single-use
assemblies is critical to quality. Quality must be built in by
ensuring proper systems are in place to minimize
particulates in the manufacturing of SUT. Relying on visual
inspection and final lot release testing does not ensure that
a visually perfect assembly is supplied to the end user.
Instituting the systems and training outlined in this
document will assist SUT suppliers (and their suppliers) in
meeting and exceeding end user expectations for control of
particulate contamination.
controls controls
“Target” “Perfect World”
Particulate Profile Particulate Profile
TIME
Continuous Improvement – Particle
Reduction and Simplification of
Investigations
SUT suppliers and end users need to cooperate to control
particles and make further reductions. Control and
improvement efforts should consider the full lifecycle of
the SUT, from manufacturing supply chains to end user
deployment in biopharmaceutical production, rather than
emphasizing a supplier vs. end user perspective.
While these practices will help with any supplier/end user
relationship, a view across the full set of suppliers and end
users would be powerful. The following best practices are
recommended:
 Continuous monitoring and trend analysis: Trending
analyses for cleanroom performance and for particle
observations made throughout the SUT lifecycle should
be performed;
 Continuous characterization: Captured particles should
be identified;
 Continuous cataloging: Particle identifications should be
added to an organized and shareable catalog wherever
possible; and
 Continuous improvement activities should be evaluated
by trending analyses.
These steps will help the industry to demonstrate that we
understand the particles that may be in SUT, and that the
systems to manage particles are in control. We must apply
continuous improvements, track results, and show that the
particle control process is improving.
21

Part X: BPSA-recommended Next
Steps
As stated in the Introduction (see page 3), this document
captures the current state of the industry in regards to SUT
and particulates.
Through the writing of this document, the group discussed
at length many items that are not fully addressed here and
which they feel need further study. These items include:
1. The industry needs a specific SUT particulate
measurement method. USP <788> is intended for final
formulation and has been adapted for SUT
components. The proper methods and requirements
need to be established for in-process SUT components,
which should be coordinated against the requirements
of USP <788> and other monographs.
2. Application-specific requirements need to be better
defined. These include final bulk product vaccines and
cell therapies that require lower levels of particles
than other routine processes more typically subject to
final sterilizing filtration to control particulates in
filled products.
3. An industry-wide catalog of particles would facilitate
investigations. Specific SUT components’ particulates
could be cataloged and referenced by final assemblers
and end users during investigations.
4. Particulate generation studies using simulated end
user processes should be conducted to verify best
handling practices.
5. A formal guide to cover all SUT Best Practices, in
addition to particulates, should be issued. This
document would address proper handling and use of
the SUT in general, and should include techniques that
minimize particle formation.
6. Supplier/End–user Quality Agreements of acceptance
criteria for the SUTs should be established. These
should utilize many of the principles mentioned in this
document and clearly document all acceptance
criteria. BPSA has created a Quality Agreement
Template that provides an example of a common
structure for Quality Agreements between suppliers
and end-users.

Figure 13 (see below) provides an example of how suppliers and end users might discuss and document their agreement. In
the figure, the red line represents the agreed-to acceptance criteria for a specific application. This may vary for each SUT and
each application.

Figure 13: Example of end user/supplier agreement for particulate acceptance criteria

USP<788> MATERIALS OF
NONE PACKAGING
ACCEPTABLE [< 3 particles/mL CONSTRUCTION ACCEPTABLE
(>25 microns)]
[<25 particles/mL
(≥10 microns)]

CLASS VI OUTSIDE
MATERIALS OF USP<788>
CONSTRUCTION [3 particles/mL
(>25 microns)]
[25 particles/mL
(≥10 microns)] ENVIRONMENT

USP<788>
FOREIGN FOREIGN
[>3 particles/mL
(>25 microns)]
[>25 particles/mL VIABLE
(≥10 microns)]
HAIR/BUG TOXIC PROCESS/PRODUCT
UNACCEPTABLE VIABLE CONTACT UNACCEPTABLE

VISIBLE SUB-VISIBLE CHEMICAL LOCATION

MAKE-UP

Indicates the agreement level for each application

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Part XI: Terms and Definitions
Active ingredient “Any component that provides pharmacological activity or other direct effect in the diagnosis, cure,
mitigation, treatment, or prevention of disease, or to affect the structure or any function of the body of
man or animals.” (per FDA)

Assembly Process equipment consisting of components or sub-assemblies (e.g. bioreactor, filling manifold, bag
assemblies, etc.) (For the purposes of this document, SUT)

Best practices Procedures that, in the opinion of this document’s contributors, are accepted or prescribed as being
most effective to achieve the reduction of particles.

Biological product “Any virus, therapeutic serum, toxin, antitoxin, or analogous product applicable to the prevention,
(protein) treatment, or cure of diseases or injury.” (per ISPE)

Cells “The fundamental unit of life. The living tissue of every organism is composed of these fundamental
living units. Unicellular organisms, such as yeast or a bacterium, perform all life functions within the
one cell. In a higher organism, a multicellular organism, entire populations of cells may be designated a
particular task.” (per ISPE)

Chemical make-up The elemental or compound consistency of the particle.

of particle

Clean room “A room in which the concentration of airborne particle is controlled to specified limits.” (per ISPE)

Component “An element within an assembly, such as a pump or a valve, on a piece of process equipment. These can
be stand-alone or used as part of a larger assembly (e.g. [SUT examples] bag, tubing, filter, connector,
etc.).” (per ISPE)

Controlled A clean room that has a controlled level of contamination that is specified by the number of particles
environment per unit volume at a specified particle size (e.g. ISO 7 or ISO 8 level controlled environments).

Embedded Fixed firmly in the surrounding material.

End user “The body upon which final possession or use of a single-use device rests.” (a.k.a. Owner-User) (per
BPE)

Endogenous Endogenous particulates are those derived from constituents of the pharmaceutical process or product.
These particulates are a result of the product itself.

Extrinsic (foreign) Not related to process contact materials and typically include rubber, metal, plastic, hair, fibers, insect
parts, and dust coming from the manufacturing environment.

Free Not physically fixed to process contact materials.

Gel "A nodule of plastic material composed of one or more of oxidized, high molecular weight, un-melted,
non-solvated, or cross-linked material of the same composition as the matrix that, for a variety of
reasons, has not blended with the matrix." (per ASTM D883)

Inherent Existing in the material as a permanent, essential, or characteristic attribute.

(formulation)

Intrinsic (material The process contact materials of the SUT. Intrinsic particles are part of the process/product like
of construction) formulation ingredients, packaging materials, etc. They are native and from the SUT itself. They are
reasonably expected to appear and sometimes are generated with the use of the SUT. These particles
are all non-viable and do not contain living organisms.

Non-viable “A particle that does not consist of, or support, one or more live microorganisms.” (per ISPE)

Packaging Non-process product contact materials used to wrap, ship and/or protect a component.

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Particle Loose mobile matter or embedded matter that is unintentionally present in/on the single-use
component/assembly and potentially may contact or may end up in the process/product fluid.

Particulate 1. Particle (see above), or

2. Particulate matter in injections and parenteral infusions consists of mobile un-dissolved
particles other than gas bubbles unintentionally present in the solutions. (USP <788>)

Process application How the SUT is used in the production of the drug product.

Process contact surface “Surfaces of process components that, under normal design operating conditions are in direct
contact with, or have the potential to come in contact with raw materials, in-process materials,
Active Pharmaceutical Ingredients (APIs), clean utilities (e.g., WFI, Pure Steam, CIP solutions,
process gases), and where there is a potential for a component (e.g., stoppers) surface to
impact drug product(s) strength, identity, safety, purity, and quality.” (per ISPE)

“Surfaces of tubing, equipment or systems that, under design operating conditions, are in
contact with, or have the potential to contact, raw materials, in-process materials, APIs, clean
utilities (e.g. WFI, CIP, Pure Steam, process gases), or components (e.g. stoppers), and where
there is a potential for the surface to affect product safety, quality, identity, strength or purity.”
(per ASME BPE)

Product contact surface “A surface that contacts raw materials, process materials, and/or product.” (per ISPE)

“Product contact surfaces that are in contact with, or have the potential to contact, product
where product is defined by the owner/user. Examples of product contact surfaces may
include the interior surfaces of bioreactors, transfer tubing, chromatography columns, vessels,
and recirculating segments of CIP systems.” (per ASME BPE)

Quantity/count of Particle The number of particles per unit surface area or volume.

Risk “Combination of the probability of occurrence of harm and the severity of that harm.” (ISO/IEC
Guide 51) (ICH Q9).

Single-use technologies “Consist of fluid path components to replace reusable stainless steel components. The most
(SUT) [also known as typical systems are made up of bag chambers, connectors, tubing, and filter capsules.” (per
single-use systems (SUS)] BPSA).

Size of particle The physical net dimension of a particle, usually equated to a spherical equivalent.

Standard (ANSI-accredited “Organizations whose primary activities are developing, coordinating, revising, amending,
or equivalent) reissuing, interpreting, or otherwise producing technical standards that are intended to
address the needs of some relatively wide base of affected users.” They include ASTM, USP,
ASME-BPE, ISO, CFR, EP, and JP. (per www.wikipedia.com)

Sub-assembly An assembled component that can be used on its own or as part of a larger assembly (e.g.,
sample device, ported bag, process manifold, etc.).

Sub-visible Difficult to detect by unaided human eye. (For the purposes of this document, sub visible will
be considered <100 microns).

Supplier “An organization or individual, internal or external to the user, associated with the supply
and/or support of products or services at any phase throughout a systems lifecycle.” (per
ISPE)

Viable “A particle that consists of, or supports, one or more live microorganisms.” (per ISPE)

Visible particle Particles > or equal to 100 microns in size. (For the purposes of this document--which is based
on specification limits and methods to detect particles--this size was agreed to.) (USP <1>)

24
Part XII: References
1. Stephen E. Langille; Particulate Matter in Injectable Drug Products. PDA J Pharm Sci Tech 2013, 67:186-200.
2. http://www.fda.gov/Safety/Recalls/ArchiveRecalls/2012/default.htm?Page=11
3. Woodcock, J. (2004) The Concept of Pharmaceutical Quality. American Pharmaceutical Review, 7 (60), pp. 10-15.
4. WHO IP (5.7) Test for Particulate Contamination.
5. http://www.aaps.org/uploadedFiles/Content/Sections_and_Groups/Focus_Groups/PABCFGnewsMay2011.pdf
6. Ankers, Michael J., Larrimore, Dan, Morton Guazzo, Dana; Parenteral Quality Control: Sterility, Pyrogen, Particulate, and
Package Integrity Testing, Marcel Dekker, 2003, p. 94.
7. Thomas A. Barber; Control of Particulate Matter Contamination in Healthcare Manufacturing. CRC Press, 1999, ISBN:
1574910728.
8. USP <790> Visible Particulates in Injections. In preparation.
9. USP <1790> Particulate Matter Determination: Visual Inspection. In preparation.
10. USP <788> Particulate Matter in Injections. USP <36>; U.S. Pharmacopeia National Formulary, 2013; pp 350 –353.
11. USP <1788> Methods for the Determination of Particulate Matter in Injections and Ophthalmic Solutions. U.S.
Pharmacopeia National Formulary, 2013. USP <36>. pp. 6225–6235.
12. USP <787> Subvisible Particulate Matter in Therapeutic Protein Injections. U.S. Pharmacopeia National Formulary. In
preparation.
13. USP <1787>Measurement of Subvisible Particulate Matter in Therapeutic Protein Injections. U.S. Pharmacopeia National
Formulary. In preparation.
14. ASTM F24-09 Standard Test Method for Measuring and Counting Particulate Contamination on Surfaces. April 2009.
15. ATM D883 Standard Terminology Relating to Plastics, www.astm.org
16. AAMI TIR42:2010 Evaluation of particulates associated with vascular medical devices.
17. USP <1> Injections. U.S. Pharmacopeia National Formulary, 2013. USP <36>. pp. 33-37.
18. EP 2.9.20 Particulate Contamination: Visible Particles. 2008; European Pharmacopoeia 8.0; 01/2008; pg. 323.
19. EP 2.9.19 Particulate Contamination: Sub-Visible Particles. European Pharmacopoeia 8.0; 04/2011; pg. 321-323.
20. JP 6.06 Foreign Insoluble Matter Test for Injections. Japanese Pharmacopoeia Sixteenth Edition.
21. JP 6.07 Insoluble Particulate Matter Test for Injections. Japanese Pharmacopoeia Sixteenth Edition.
22. Regina M. Malczewski, Csilla Kollar, Tubing Particulates, Part 1: From USP <788> to an Extraction Connection. Dow Corning
White Paper 52-1139-01.
23. Akers, Michael J. et al., 2002, Formulation Development and Protein Dosage Forms, Pharmaceutical Biotechnology, Volume
14.
24. AAMI TIR 42:2010, Evaluation of Particulates Associated with Vascular Medical Devices.
25. AAMI, Technical Information Report: Evaluation of particulates associated with vascular medical devices, AAMI
TIR42:2010, Page 2.
26. Lawrence X. Yu, Ph.D., Director for Science Office of Generic Drugs, Food and Drug Administration, Quality by Design for
Orally Inhaled Drug Products presentation, March 2009.
27. Patrick Evard, Particulates in Single-Use Assemblies: Definitions, Risks and Methods to Control Them Presentation, July
2013.
28. www.FDA.gov.
29. www.ispe.org/glossary.

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DISCLAIMER
The information in this document is intended to capture the current state of the Single-Use-Technology Industry in regards to
Particulate Control, Testing and Evaluation. The material presented herein is intended to help characterize levels and types of
particles, as well as to provide methods to assure minimal levels of particulate in SUT. This information is offered in good faith
and supported by the expertise of its contributors. However, BPSA, its members, and contributors do not assume any
responsibility or obligation for the reader’s compliance to the content of this document. This is not a standard, but a set of
recommendations. Manufacturers, suppliers and end users should consult with their own legal and technical advisors relative
to their SUT use and participation.

About BPSA
The Bio-Process Systems Alliance (BPSA) was formed in 2005 as an industry-led corporate member trade association
dedicated to encouraging and accelerating the adoption of single-use manufacturing technologies used in the production of
biopharmaceuticals and vaccines. BPSA facilitates education, sharing of best practices, development of consensus guides and
business-to-business networking opportunities among its member company employees.

For more information about BPSA, visit www.bpsalliance.org.

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