20 20

RAW MATERIAL
PARTICLES + COMPONENT SUB-ASSEMBLY
PARTICLES + PARTICLES +
ASSEMBLY
PARTICLES + TRANSFER + UNPACKAGING + PREPARATION + USE*

SUPPLIER SIDE END USER SIDE

2020 RECOMMENDATIONS FOR

TESTING, EVALUATION, AND CONTROL
OF PARTICULATES FROM SINGLE-USE
PROCESS EQUIPMENT
2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
FROM SINGLE-USE PROCESS EQUIPMENT

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 2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
FROM SINGLE-USE PROCESS EQUIPMENT

Published by:
Bio-Process Systems Alliance (BPSA)
1400 Crystal Drive
Arlington, VA 22202

www.bpsalliance.org

2020 AUTHORS
Emily Alkandry, Saint-Gobain Life Sciences
Ken Davis, Nordson Medical
Patrick Evrard, Pall Biotech
Jayanthi Grebin, Colder Products Company
Jay Harp, Avantor
Eric Isberg, Savillex
Ernie Jenness, MilliporeSigma
Csilla Kollar, DuPont de Nemours Inc.
Tim Korwan, Avantor
Mark Petrich, Merck & Co., Inc.
Chuck Raye, MilliporeSigma
Sebastian Selzer, Eppendorf Bioprocess
Heidi Sinkovich, Meissner Filtration Products
Kirsten Strahlendorf, Sanofi Pasteur
Klaus Wormuth, Sartorius Stedim Biotech
James Dean Vogel, The BioProcess Institute
Maureen Eustis, The BioProcess Institute

2014 AUTHORS and PARTICULATE COMMITTEE CONTRIBUTORS

Ken Davis, Nordson Medical, Patrick Evrard, Pall Biotech, Eric Isberg, Savillex, Ernie Jenness, EMD Millipore
Mike Johnson, Entegris, Csilla Kollar, DuPont de Nemours Inc., Mark A. Petrich, Merck & Co., Inc.
Helene Pora, Pall Biotech, John Stover, AdvantaPure/New Age Industries, Kirsten Strahlendorf, Sanofi Pasteur
James Dean Vogel, The BioProcess Institute, Maureen Eustis, The BioProcess Institute

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 2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
FROM SINGLE-USE PROCESS EQUIPMENT

Executive Summary
The tremendous growth rate of drug products based on biopharmaceutics is driving a wide variety of innovations in the manufacturing
processes for biopharmaceuticals. Rapidly increasing adoption rates for single-use process equipment continue since single-use systems
show significant advantages in flexibility, cost and performance compared to multiple-use stainless steel systems. However, one large
concern in the adoption of single-use systems is particulate matter.
Since the first BPSA paper on particulates in 2014, increased awareness of the issue of particulates in single-use process equipment has
driven substantial improvements in single-use cleanliness with respect to particulate matter. Single-use suppliers increasingly produce
components and assemblies in qualified cleanroom environments under greater process control, reducing the overall levels of
particulates in single-use systems. In addition, end users are taking more care to maintain cleanliness in the application of single-use
systems during biopharmaceuticals manufacturing.
This paper provides best practices and guidance on how to test, evaluate and control particulate matter in the manufacture and
application of single-use processing equipment. While current manufacturing of single-use systems is not particle-free, this paper shows
how the risks associated with particulates can be dramatically reduced.
There are three factors in a risk analysis for particulates: severity, probability, and detectability. Particulates in drug products present a
risk to patient safety. Certainly, in applications of single-use equipment downstream of final filters (for example in final filling, vaccine
production and cell therapies), the probability of particulates detaching from the surfaces of single-use equipment and contaminating
the final drug product are much higher than upstream of final filters, where sterile filtration remove particulates greater than 0.2
microns. The cleanliness of single-use process equipment must meet the demands of the application, and this paper gives guidance on
how to meet these demands.
Prevention of particulates during the manufacture of single-use process equipment first requires quality by design: consideration of
particulates in the design of the single-use component and assembly, sourcing of clean components, and in the choice of the single-use
manufacturing processes. Overall, cleanroom specifications, control of the cleanliness of the manufacturing equipment, and, in
particular, the hygiene and gowning of operators, are all factors required for successful production of single-use systems with
consistently low particulate levels.
Another key in reducing risk from particulates is improving detectability. First and foremost is a rigorous visual inspection process for
single-use components and assemblies by both the supplier and end user. But, as described in detail in this paper, the detection
probability in visual inspection is reduced by the size, complexity and limited transparency of single-use systems. Often, destructive
sampling and testing, in the form of liquid extraction of the particulates from the surfaces of the single-use assemblies, is required to
more completely assess particulate levels and determine particulate risk. Critical factors are an effective extraction method, along with
effective particle measurement tools for particle counting, sizing and identification. The information from destructive testing
determines manufacturing process capability and drives continuous improvement efforts to reduce particulate levels.
Finally, communication between suppliers and end users is crucial. End users need to understand the risks they are taking in applying
single-use systems, and suppliers must give end users the information needed to meet the required quality standards with respect to
particulate cleanliness by demonstrating their manufacturing and particulate control capabilities, as well as their understanding of the
potential risks posed by particulates. This challenge becomes more important as single-use systems find increasing application after
the last filtration step, in final filling, vaccine, and cell and gene therapy applications.

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 2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
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Table of Contents
Topic Page
Part I: Introduction 4
Part II: Particle Definition & Classification 5
Part III: Risk 6
Part IV: Particle Detection and Characterization 12
Part V: Particle Inspection & Quantification 14
Part VI: Control of SUT Manufacturing Process 21
Part VII: Control of Biopharmaceutical Manufacturing 24
Part VIII: Deviation Response/Mitigation Plans 25
Part IX: Summary & Conclusion 27
Part X: BPSA-Recommended Next Steps 29
Part XI: Terms & Definitions 31
Part XII: References 33

Figures Page
Figure 1: Classification of particulate matter risks in the manufacturing
and use of biopharmaceuticals 7
Figure 2: Potential particle locations 8
Figure 3: Risk from particulate matter by category 9
Figure 4: Fishbone diagram of potential particle sources 10
Figure 5: Sources of possible particle contamination 10
Figure 6: Potential contributors to particle levels in SUT 11
Figure 7: Particle load can be reduced using best practices 11
Figure 8: Potential particle sources in SUT 13
Figure 9: Particle size classifications (not to scale) 14
Figure 10: Approximate gap in particle detectability between visual inspection and Method 1
(light obscuration) 15
Figure 11: TAPPI size estimation chart 17
Figure 12: ISO 7 cleanroom particle counts by week 23
Figure 13: Use of controls over time to reduce particulates 28
Figure 14: Example of end user/supplier agreement for particulate acceptance criteria 30

Tables Page
Table 1: Conversion of TAPPI areas into particle diameter 17
Table 2: Comparison of methods used in USP <788> 19
Table 3: Potential sources of particulates contributed by end user single-use operations 24

Appendices Page
Appendix A: BPSA User Requirements Template 34

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 2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
FROM SINGLE-USE PROCESS EQUIPMENT

2020 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 globally facilitate the development
and manufacturing of biopharmaceuticals through the implementation of robust, safe and sustainable Single-Use
Technologies. Corporate members include plastic bioprocessing equipment suppliers, service providers and end 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, end users and
regulators to safeguard the quality of drugs produced with single-use process technologies.

Intent and Scope “The goal of end users, regulators, and standards-setting
This document provides recommendations to suppliers and end organizations should be to minimize particulates in drug
users in the single-use technology (SUT) industry regarding products, without placing unnecessary expectations on
particulate testing, evaluation, and control. The practices suppliers for minimal safety gains. Improving the
presented here are intended to help characterize levels and types manufacturing quality will reduce the risk of harm to
of particles, as well as to provide methods to assure minimal patients from particle contamination.” (Ref. 4)
levels of particulate in SUT.
In summary, for SUT fluid paths, fewer particles/particulate is
better. While zero is the goal, it is not an achievable specification.
Background This means the SUT industry must continue to strive for fewer
The BPSA recognized a need in the SUT industry and started particles and fewer types of particles in the SUT.
discussions about particulates at its 2012 and 2013 International
Single-Use Summits. In 2014, the original edition of this paper In this document, BPSA members provide a view to help the SUT
was published which captured the challenges and best practices community navigate this complex topic throughout the lifecycle
faced by suppliers and end users as they pertain to particulates in of the SUT.
SUT. The content significantly influenced the industry by This document helps clarify:
providing a basis of discussion for testing, evaluation, and control
of particulates. The previous effort also influenced discussions  Why are particles a concern when using SUT?
which occurred in standards organizations resulting in a  Why are particulates in SUT a risk to the:
particulate requirement added to the 2016 ASME BPE Standard o Patient who is being treated with the drug product?
for single-use, and also initiation of ASTM work groups. Efforts o Purity of the drug substance or drug product?
such as issuances of PDA Technical Report 79 Particulate Matter o Yield and product quality of the process fluid?
Control in Difficult to Inspect Parenterals, USP <790> Visible  What factors are key to assessing particulate risks from
Particles in Injections, and <1790> Visible Inspection of Injections SUT?
(Refs. 1, 2, and 3) took place after 2014 and are considered in this o What is the potential severity of the harm from
update. Furthermore, the advent of cell and gene therapy, post- particulates in SUT?
2014, accelerated the need for particulate testing, evaluation, o What is the probability a particle in an SUT will reach the
and control for SUT items. This document builds on the content patient?
contained in the 2014 edition and includes a significant amount o Are filters applied downstream of the SUT?
of new information to better guide the SUT community.  How can you improve the detectability of particulates in
SUT?
Industry Practice o What particulates are realistically detectable in a visual
BPSA members, and the industry as a whole, recognize that inspection?
achieving zero particles is not physically possible for process o How can you do a quantification using extraction and
equipment used in filling of injectable drug products. Substantial particle counting?
progress has been made by leaders in the industry since 2014 and  How do you distinguish levels of particle risk based on
the goal is to continue to reduce the levels, thus the risk, of location in the biopharmaceutical manufacturing process?
particles as much as possible. We continue to add single-use  How do you control and minimize particulates during the
technology suppliers to the goal stated by an FDA author: manufacture of SUT?
o How are particulates prevented?

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 2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
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o How is particulate cleanliness properly managed in a Intrinsic particulates come from sources within the
cleanroom? biopharmaceutical manufacturing process, such as the contact
o How do you address situations where particulates are materials (SUT) and the formulation and processing ingredients
found in, on, or embedded within SUT? (buffers, stabilizers, etc.). Intrinsic particles from SUT materials
o How are particulates measured for process capability and components may be created during SUT manufacturing
and continuous improvement efforts? (cutting, welding, assembly, etc.) or created during SUT
application (mixing processes, etc.). These particles are expected
This paper's audience includes those who are new to SUT as well
or known particles, since the materials comprising the SUT are
as those who are already familiar with the technologies. It is
known and well characterized. Also, these particles are most
anticipated that the document will provide a base for both groups
likely non-viable since they typically do not contain living
to gain a common understanding of this complicated topic.
organisms.
This document is also intended to help both the end user and the
Inherent or endogenous particulates are a type of intrinsic
supplier. It can help the end user convey their specific
particle known to be associated with specific drug product
requirements to the supplier. In turn, suppliers can use the
formulations. For example, protein related products may contain
document to demonstrate what they are capable of providing to
inherent protein related agglomerates.
the end user.
Extrinsic particulates come from sources external to the
Definitions of terms are provided at the end of this document to
biopharmaceutical manufacturing process, and so are considered
assist in the ongoing dialog among end users, suppliers, and
foreign to the process. The extrinsic particles category most
regulators.
commonly includes, but is not limited to, this list of potential
particles coming from the SUT manufacturing environment or
Part II: Particle Definition & Classification from the biopharmaceutical manufacturing environment:
Particle Definition  hair or skin flakes from human operators
In general usage, the term particle is usually defined as a minute  non-process related fibers (from clothing, wipes, etc.)
portion, piece, fragment, or amount of matter. Technical  cellulose (paper, cardboard)
organizations such as AAMI, ASME, ASTM and others formally  rubber, metal, glass, plastic, dust
define the term particle in various but overall similar terms. In the  insect parts
US Pharmacopoeia monographs concerning parenteral injectable  microorganisms, pollen, bioburden
drug products and infusions, the terms particles, particulates and Viable particulates are a subset of extrinsic particulates which
particulate matter are considered equivalent and do not have contain one or more microorganisms. Particulate viability can be
different meanings (USP <1790>) (Ref. 3), and particulate matter mitigated with sterilization, provided the validated bioburden
is defined in the USP <788> (Ref. 5) as: levels are maintained within controlled levels. Residuals from the
“Extraneous mobile undissolved particles, other than elevated bioburden may be toxic, such as endotoxins. Endotoxins
gas bubbles, unintentionally present in solutions.” are not particulate matter, but are considered to be biologically
active pyrogens even at low concentrations.
In the production of biopharmaceuticals, the typical matrices are
aqueous-based, and detectable particulates are solids or Known particulates are chemically/physically identified particles
immiscible liquids. The BPSA definition of particles includes SUT which are usually tracked to original sources. Such particles can
specific language: be trended and understood within the risk analysis related to the
single-use technology lifecycle (Part III).
“A particle is loose mobile or embedded matter that
is unintentionally present in/on the single-use Technically difficult to avoid particulates are a subset of known
component/assembly and potentially may contact particulates which are difficult to avoid using current SUT
or may end up in the process/product fluid.” manufacturing technologies. Typically, these are particulates
such as intrinsic particulates from the known plastic materials of
Classification Based on Particle Sources
construction of the SUT, and extrinsic particulates such as textile
Particles may originate from many sources. Particles found in or
fibers from clothing and gowning, and particulates from paper or
on SUT are classified by the type of source (intrinsic/extrinsic), by
cardboard packaging. These are often expected to appear at
the level of identification and ability to identify the source
some level in SUT, and knowledge of these types of particulates
(known/unknown), and by biological activity (viable/non-viable).
should be communicated from the supplier to the end user.
Particles found upon application of SUT in biopharmaceutical
Unknown particulates are of the MOST CONCERN. These
processes might be defined somewhat differently depending
particles are difficult or impossible to chemically/physically
upon how the SUT is applied. For example, some particles
identify, and cause significant doubts regarding quality systems
considered extrinsic in closed systems may be considered
and environmental controls, since finding the original sources
intrinsic if SUT are used in open manipulations.
may not be possible.

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These definitions are critical to understand the risk of particles. or biological product containers. From a quality risk management
Of all these categories--extrinsic, reactive and unknown particles- perspective, particulates are a potential hazard which can
-are the worst-case scenario. As described in the next section, potentially cause harm to the patient receiving the drug product.
these definitions and classifications of particulates are important The potential harm from particulates not only includes possible
for understanding the issue of risk from particulates from SUT in damage to patient health, but also loss of drug product quality,
biopharmaceutical manufacturing. efficacy, and availability. (Ref. 6) One of the top 3 reasons for drug
product recalls is the presence of particulates.
Part III: Risk Risk is defined as the probability of occurrence of harm multiplied
There are a wide variety of stakeholders in the manufacturing and by the severity of that harm. The severity of the harm of
use of biopharmaceuticals made with SUT: particulates to the patient depends on a large number of factors,
 Single-use process equipment suppliers (and their suppliers) not all of which are well understood (Ref. 8), including:
 Raw materials suppliers (for example: salts, excipients)  patient factors (state of immune system, age, etc.)
 Biopharmaceutical manufacturers  frequency of administration (once/annually vs. regularly)
 Medical practitioners  route of administration
 Regulatory agencies  use of filtration at the point of administration
 the volume administered/dosage
....and, of course, the most important stakeholder is the patient  particle size and their risk to the patient
receiving the drug product.  particle type, source and amount
According to ICH Q9, the primary principle of quality risk A PDA “Industry Perspective” paper on patient safety risk claims
management is that “the evaluation of the risk to quality should that:
be based on scientific knowledge and ultimately link to the
protection of the patient.” (Ref. 6) But it is important to “a gap exists between the observation of small quantities
understand that all stakeholders may not necessarily agree on of particles in injectable pharmaceutical products and the
the degree of risk from particulates. patient documented safety concerns resulting from
inadvertent administration of particles to patients...the
Hence, there is understandably a high sensitivity to particulates, existing data suggest the overall risk to patients is generally
regardless of the level of scientific knowledge and regardless of low and the benefit of these treatments is generally
where particulates are found within the lifecycle of significant”. (Ref. 8)
biopharmaceuticals made with SUT.
Regardless of this perspective, other stakeholders
Regulatory requirements for final drug Visible particles are a visible have a different assessment of the risk of particles.
products (pharmacopoeia) and for quality indicator, and even
biopharmaceutical manufacturing (cGMPs) In most cases where regulatory requirements are
the presence of only one carefully followed, the probability of occurrence
come first and foremost. Regulatory
particle suggests to some of particulates is low and the severity of harm is
requirements very clearly spell out the
stakeholders that a process limited, since the patient receives, at most, very
expectations for equipment, their surfaces,
and any particulate that may be contained
might be out of control. small doses of sub-visible and visible particulates
(Ref. 7) from drug products. (Ref. 4)
within the equipment. US FDA Drug cGMP,
21 CFR 211.65 states: Risk knowledge helps to focus improvement
“Equipment shall be constructed so that surfaces that efforts on the most critical aspects of the process. Risk awareness
contact components, in-process materials, or drug products is important during process development, technology transfer,
shall not be reactive, additive, or absorptive so as to alter the monitoring and control.
safety, identity, strength, quality, or purity of the drug
product beyond the official or other established
requirements.”
US FDA cGMP for Biologicals, 21 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
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 2020 RECOMMENDATIONS FOR TESTING, EVALUATION, AND CONTROL OF PARTICULATES
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Figure 1: Classification of particulate matter risks in the manufacturing and use of biopharmaceuticals

Particulate
Matter

Concern: Concern: Concern:

Process Performance Product Quality & Efficacy Patient Safety
(yield) (purity) (health)

Assess Potential Assess Potential Assess Potential

Interactions Interactions Interactions
Process Performance Product Medical Risk
(yield) (protein/vaccine/cells) (human)

In addition to the overriding concerns regarding particulates in Risks in process applications of SUT upstream of final filters
the final drug product, particulates present within the (Lower Risk)
biopharmaceutical manufacturing process stream also present For other applications of SUT, properly validated application of
risks to process performance and ultimate drug product quality. final 0.2 micron filters will essentially eliminate risk for direct
Since the application of validated sterilizing grade filters (0.2 transfer of particulate matter from SUT upstream of the final
micron) effectively removes all particulates greater than 0.2 filter into the final drug product. However, in applications
microns, overall there are two general categories of risk for upstream of final filters, particles from SUT potentially present
biopharmaceutical manufacturing processes using SUT: some risk to process yield and process fluid purity. In these cases,
a risk assessment should consider potential “secondary” effects:
Higher Risk: Processes which occur downstream of final filters
1) the potential influence of particles on biochemical
Lower Risk: Processes which occur upstream of final filters
reactions;
Risks in process applications of SUT downstream of final filters 2) the potential risks from any leachables from particles on
(Higher Risk) the process fluid;
In general, applications of SUT after the last filtration step in a 3) potential risks from microorganisms carried by particles
biopharmaceutical manufacturing process present, by far, the into the process fluid; and
highest potential risk for transfer of particulate matter directly 4) potential risks to increased protein aggregation nucleated
from the surfaces of the single-use system into the final drug by particles on the quality of the process fluid.
product. Falling into the same category of higher risk are SUT
Particulates may interact with cells
process applications for products which cannot be filtered, such
In Cell and Gene Therapy (CGT), particulate matter may interact
as some vaccines and most cell therapy products.
with cells by changing cellular adhesion (changing cell-cell
For such higher risk applications of SUT, a high level of attention interactions), or being taken up by the cell (phagocytosis). (Ref.
must be given to understanding and controlling the level of 9)
particulate matter, especially on the fluid contacting surfaces of
Chemicals may leach from particulates
the SUT. In these cases, a holistic particle risk analysis for the final
In assessing risks from leachables from particles in a process fluid
drug product not only involves a careful assessment of the
upstream of final filters, the type of particle and the
potential risk contributions from the SUT, but also the potential
concentration of particles in the process fluid should be
risk contributions from final containers (vials, ampules, syringes,
considered. Intrinsic particles composed of the materials of
rubber stoppers, etc.) and any excipients added into the final
construction of the SUT likely present the lowest risk. Even with
formulation after the last filtration step.
large amounts of such particles, the particles are of known
materials, extractable data usually exists for these materials, and

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the increase in surface area due to such particles is usually a rinsed with cleaning chemicals and high purity water. The
negligible contribution to the overall surface area of the SUT. objective of these cleaning processes is to remove soil and
Intrinsic particles from SUT materials of construction are low risk environmental contaminants. This includes reducing the levels of
to process fluids upstream of final filters. Regardless, intrinsic surface particulates.
particulate levels should be minimized as practically possible.
During biopharmaceutical manufacturing, SUTs usually do not
On the other hand, for extrinsic particles, which can be wide undergo a final rinsing step. Particulates generated during SUT
ranging in chemical composition, extractables/leachables data manufacture, or from the SUT manufacturing environment, may
usually do not exist. However, estimations of the potential remain on the SUT’s fluid contact surfaces.
amount of leachables based upon worst case scenarios can help
Particles may be located on the inside, outside, or embedded
in assessment of risk. For typical plastics and textile fibers,
within SUT
additive concentrations range from approximately 1 to 10%. As
Particles may be present on the surfaces of, or embedded within,
an example of worst case scenario calculation, consider a cotton
the materials of SUT components and assemblies. (Figure 2)
fiber from clothing: 2.5 mm long, 20 nanometers wide, about 1.2
micrograms of material. If the additive concentration of the fiber Figure 2: Potential particle locations
is assumed to be 10%, and all additives are completely leached
from the fiber into a liter of process fluid, the leachables
concentration would be in the order of 0.1 microgram/liter (0.1
ppb). In this case, such low levels of leachables from low Exterior
concentrations of particulates likely present low risk to product SUT Surface Process/Product
quality. Regardless, extrinsic particulate levels should be Contact Surface
minimized, especially since the perception of risk is higher than (interior)
that for intrinsic particulates.
Viable microorganisms may be attached to particulates
An additional risk to process fluids are extrinsic particles
potentially carrying viable microorganisms such as bacteria. In
this category are hair and skin flakes from human operators. For embedded within
example, up to a million bacteria are found in a gram of human component
hair. (Ref.10) A single hair fiber 6.5 mm long and weighing about
20 micrograms can potentially carry around 20 bacteria into the Process/Product Contact Surface: This is the surface of the SUT
process fluid. While a single hair likely presents a low risk to which comes into direct contact with process fluids, drug
increased bioburden in a SUT, the perception of risk is very high substance or drug product, depending upon SUT application.
for particulates which potentially contain microorganisms such as Liquids wet these surfaces, and any particles attached to these
hair, skin flakes, etc. surfaces could be released directly into the fluid. In terms of risk,
particles on the process/product contact surface are of highest
Particulates may nucleate protein aggregates risk for ending up in the fluid. Therefore, particles on these
Finally, a risk assessment of particles in process fluids upstream surfaces are the primary focus of this paper.
of final filters should consider the possibility that particles could
potentially increase denaturation and aggregation of proteins. Exterior SUT Surface: Particles present on the outside of the SUT
The research literature shows that particles can potentially are usually a low risk to most processes. Most SUT processes are
nucleate protein aggregates, and steel particles are more closed and routinely not exposed to the exterior of the SUT.
problematic than cellulose particles. (Refs. 11 and 12) On the Exterior particles, however, may affect other adjacent processes,
other hand, this factor is in addition to a long list of other cleanroom, or isolator operations and should be addressed
chemical and physical stress factors which can denature proteins. appropriately.
Upstream of final filters, most biopharmaceutical manufacturing Embedded: Embedded particles in SUT components and
processes consist of many purification steps using assemblies may create a low risk to the process depending on
chromatography and filtration, which substantially reduce the whether they are fully encapsulated or exposed to the process
risks for the presence of denatured proteins in final drug contact surface. The size and shape of the embedded particle
products. could impact mechanical integrity. The chemical composition of
SUT-specific Risk Considerations the particle could impact potential leachables. Definitions of
SUTs are not usually rinsed prior to use. SUTs are often employed embedded particles in polymers are given in ASTM D883-20. (Ref.
to provide closed-system processing with ready-to-use sterilized 13) Gel particles are nodules of polymer material, unmixed or un-
components and assemblies. Traditional pharmaceutical multi- melted, that appear as “fish-eye” shaped defects in a plastic. In
use equipment (e.g., stainless steel) is usually cleaned before extrusion or injection molding, gels could oxidize and become
sterilization. During these cleaning processes, the surfaces are yellow, brown or black depending upon the level of oxidation. Gel
particulates should be considered intrinsic and difficult to avoid
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during the plastic production process, and are minimized but not Major classifications for SUT particle risk are:
completely eliminated in well controlled extrusion/molding Process application (the application or use of the SUT):
processes. In addition, foreign particulates, which are extrinsic to Upstream (bioreactors, etc.)
the plastic extrusion/molding process, may also be embedded in Mixing
plastic materials used in SUT. As published in a recent paper (Ref. Storage
14), the risk of significant unknown leachables from any type of Transfer
embedded particles is low due to small size and usually low Filling systems
amounts of materials. In addition, the paper shows that even Location of particle in or on the SUT
large gel particles have minimal impact on the mechanical Product contact surface (usually the interior of
integrity of films used to make SUT bioprocess bags. the SUT)
Exterior of the SUT
Particles may be generated by moving parts in SUT
Moving parts within SUT, such as mixing systems within stirred Embedded in the SUT material
tank reactors, tubing, and peristaltic pumps, can potentially Particle characteristics
generate particulates. Moving plastic components may rub Intrinsic or extrinsic
against each other and generate wear particles or potentially Viable or non-viable
generate protein aggregates in mAb formulations. (Ref. 15) Also, Size: Visible or sub-visible
as is well known, peristaltic pumping of tubing may generate Morphology and color
wear particles. (Ref. 16) Chemical composition of the particle
Count (total number) of the particles
Summary of potential particulate risks from SUT
In summary, the characteristics and potential risks presented by Figure 3 below summarizes the categories of particle risk to the
the SUT to the biomanufacturing process can be viewed in terms biomanufacturing process and illustrates the risk continuum that
of where particles are found and the types of particles present in should be assessed for each specific SUT application.
the SUT.

Figure 3: Risk from particulate matter by category

LOW HIGH
Packaging RISK Process
Location of Particles Contact
Exterior Embedded Surface
LOW
RISK
HIGH Location
Process Application Media Final Fill
Process
LOW HIGH Application
RISK
Size of Particle Small Large
Visible Size
LOW HIGH
RISK Quantity
Quantity of Particle None Many
Chemical
Composition
LOW HIGH
Chemical Composition Intrinsic Unreactive Known RISK Extrinsic Reactive Unknown
of Particle (Material of Construction)

Single-Use Technology Lifecycle Each step of the SUT’s lifecycle can contribute to the potential for
Ultimately, the level of particles needs to be suitable for the additional particulates. A proper Particulate Management
intended use. Particle level data are necessary for suppliers and Program will minimize these contributions and keep the levels as
end users to assess process risk posed by the SUT. low as possible.

Ideally, the SUT would be assembled in a “clean build” manner A common approach of Particulate Management Programs
with the objective of continuous reduction of the levels of utilizes a fishbone diagram to evaluate five potential sources for
particles present. The objective of this document is to help begin each step of the SUT lifecycle to identify potential root causes of
to define the requirements that SUT end users have, and allow particulate contamination. (Figure 4)
SUT suppliers to show how they can meet or strive to meet those
requirements.
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Figure 4: Fishbone diagram of potential particle sources

ma me pe
ch ine th op
od le
s

a terials n ment
m iro
env

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 5.
Figure 5: Sources of possible particle contamination

ma me pe
ch ine tho op
ds le
ma me pe
FILTER ch ine tho op
rials nt ds le
mate onme
envir
rials nt
TUBING mate n vironme
e
CONNECTORS
ma me pe
ch ine tho op
ds le
SAMPLE
rials nt
mate onme
envir ma
ch me pe
ine tho op
ds le

rials nt PRODUCT BAG

mate onme
envir

ma me pe
ch ine tho op
ds le

rials nt
mate n vironme
e

Each step in the SUT lifecycle also contributes potential connectors, O-ring, filter membrane);
particulate risks. This increases the potential risks of particles  Assembly of the sub-assembly (bag with ports, filter capsule,
even further: connector);
 Assembly of the final assembly (bag assembly, transfer
Supplier: assembly, filling assembly);
 Raw materials of the components (resins, compounds);  Packaging; and
 Preparation of equipment (cleaning and/or sanitization) for  Transportation and handling
manufacturing components or assemblies;
 Fabrication of each individual component. (bag film, tubing,

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End user investigation. When applied correctly, these trends are like
 Receipt; instruments on a plane’s dashboard, indicating to the pilot (the
 Quality inspection; supplier or end user) where the particle levels are, and where
 Storage; they are most likely headed. The pilot now has information to
 Transfer to the production area; react and make appropriate corrections for better management
 Removal of packaging (may occur at various stages); of the levels of particles.
 Preparation (e.g., rinsing, autoclaving); Two phases in the lifecycle of an SUT need to be considered and
 Use; and integrated when analyzing particle generation and control. The
 Disposal first phase includes the supplier’s sources, manufacturing and
assembly, packaging, and shipping. The second phase includes
The particle levels should be tracked and trended in order to the end user receiving, handling, and using the SUT. Particles may
monitor performance and measure the effect of improvements be introduced to, and generated within, the SUT in both phases.
or process upsets. What is critical, especially for visible particles,
is the notion of capability: it is essential for the end user to Figure 6 summarizes the sources of particles in SUT. The left side
understand what its capability is in terms of visible particles, and illustrates the possible particulate contribution from the supplier
have the monitoring tools to follow it. If a significant deviation is during manufacturing (machine, methods, people, materials, and
observed versus its capability, this triggers an alarm and requires environment). The right side shows the end user particulate
contribution prior to or during use.

Figure 6: Potential contributors to particle levels in SUT

RAW MATERIAL COMPONENT SUB-ASSEMBLY ASSEMBLY

PARTICLES + PARTICLES + PARTICLES + PARTICLES + TRANSFER + UNPACKAGING + PREPARATION + USE*

SUPPLIER SIDE END USER SIDE

* The use of a filter by the End User decreases the number of particles.

Figure 7 illustrates how best practices can be applied to reduce reduce risk by decreasing particle levels relative to the product's
particle levels in SUT assemblies. These best practices will baseline.

Figure 7: Particle load can be reduced using best practices

traditional
PARTICLE LOAD

clean build

FILTER end user

remediation
via filter

PROCESS STEP

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Chain of Responsibility: Assembler- To paraphrase risk from particulates in biopharmaceutical

manufacturing:
Manufacturer-Manufacturer’s Suppliers
While particulate control is the responsibility of each stage of the The goal of suppliers, end users, regulators, and standards-setting
supply chain, it is ultimately the final assembly provider who organizations should be to minimize particulates in drug products,
needs to minimize particulates the most. The final assembly without placing unnecessary expectations for minimal
provider is responsible for ensuring that the assembly meets safety/quality gains. (Ref. 4)
requirements and contains minimal particulate. Ultimately, this will reduce the potential risks to patients from
In addition to controlling its own environment and implementing particles. But there is also another very important aspect of risk
procedures to eliminate particulate in its operation, the final from particulates in SUT. Not only is the probability of occurrence
assembly provider needs to understand the particulate risks that and severity of risk important, but also the detectability. (Ref. 6)
can come from each component, and hold each component The next section covers the significant challenges regarding the
provider to the equivalent level of particulate controls used by detection and characterization of particulates in SUT.
the final assembly provider.
Each component provider should understand where particulate Part IV: Particle Detection and
can come from in the operation and what type of particulate this Characterization
is likely to be. It is a best practice to have quality agreements,
such as in the BPSA Quality Agreement Template, in place that In the pharmaceutical industry, analysis of particulate matter in
define acceptance criteria, as well as suggesting inspection drug products benefits from instrumentation, lab skills, process
procedures to be put in place to identify and remove particulate knowledge, and a scientific investigative approach. (Ref. 18)
before component packaging. Similarly, identification of the sources of particles within the
manufacturing processes for SUT often requires not only
Each level of the supply chain needs to assess any packaging instrumentation for physical and chemical analysis and
materials involved with their component and the packaging of corresponding lab skills, but also keen investigative skills and a
each component. As with the components, this packaging should good understanding of manufacturing processes for SUT.
contain a minimal level of particulate. It should also be
determined whether components will need to be multi-bagged, Typical sources of particles are materials from SUT components,
especially if they are going to end up in a cleanroom for assembly. bags, tubing, and connectors, machines and tools, cleanroom
supplies, operators and clothing, and paper and packaging
Risk Summary materials. For example, particles generated in cutting and
ICH Q9 and PDA Technical Report 54 give significant guidance on welding processes during SUT manufacturing may show unique
how to approach quality risk management in pharmaceuticals physical shapes. Processes which heat materials may cause
production (Refs. 6 and 17). These recommendations emphasize oxidation, which is likely detected in the chemical analysis of the
that quality should be maintained throughout the entire product particle. A recent study shows the distribution of visible
lifecycle, and provide a proactive means to identify and control particulates found in a sampling of single-use bags currently on
potential quality issues. The risk assessment (risk identification, the market. (Ref. 19)
analysis, and evaluation) is combined with risk control (risk Particle Physical Characteristics
reduction, risk acceptance) and, importantly, communication of Particles are usually observed under an optical microscope using
risks to all stakeholders. various lighting conditions, or under a scanning electron
Communication of particulate risks between suppliers and end microscope (SEM). Here are the key physical characteristics of
users of SUT is essential. Particulate-free SUT remain the ultimate particles important for classification and determination of
goal but this is currently unachievable. original sources:
Particle Description
Characteristic
Size sub-visible, visible (see definition
below)
Shape round, angular, fiber, irregular, rod-
shape, twisted
Appearance transparent, turbid, opaque, color,
polarized
Texture smooth, rough, irregular
Hardness soft, viscous, deformable, elastic,
brittle

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Particle Chemical Characteristics Viable Particles (bioburden)

Particles can be chemically identified and classified using a variety Vegetative microbial cells or spores capable of germination and
of methods, most commonly infrared microscopy (micro-FTIR), generating live cells may be considered viable particles and can
Raman microscopy, or scanning electron microscopy with x-ray often be identified based upon morphology (size/shape). Viable
detectors (SEM-EDX). The key to effective chemical identification particles may be introduced into the SUT from the environment
is an understanding of the strengths and limitations of the chosen and personnel. Even low levels of irradiation can provide a
measurement method, along with reference libraries, and also significant level of microbial control, and sterilization ensures the
libraries of spectra from the materials, processes and absence of viable cells (viable particles). However, their presence
environment around the SUT manufacturing operations. and potential byproducts (e.g. endotoxin) may still contribute
risks to pharmaceutical products produced in SUT. Endotoxins are
not particulate matter, but are considered to be biologically
active pyrogens at low concentrations. SUT manufacturers offer
“pyrogen-free” SUT. (Ref. 19) See Figure 8 below.

Figure 8: Potential particle sources in SUT

EXTRINSIC
Other
(protein, metal, unknown, etc.)
INTRINSIC
Plastic Particles
(SUS Materials of Construction –
INTRINSIC technically difficult to avoid) EXTRINSIC
Cellulose Particles
(SUT) (paper/cardboard)
EXTRINSIC
(Known and Unknown EXTRINSIC
Sources) Textile Fibers
(cotton, dacron)

Particle Size and Visual Detectability a statistical approach with a specified acceptable quality limit
The 2014 BPSA suggestion was that the 100 micron size be the (AQL). USP <1790> describes the typical capability of a visual
benchmark for differentiating between “visible” and “sub-visible” inspection process for a drug product: 100 micron particles are
particles as depicted in Figure 9. However, 100 micron particles detected with a 40% probability, 150 micron particles are
are not always detectable in a visual inspection by the unaided detected with a 70% probability, and 200 micron particles are
eye. detected with a 95% probability. (Ref. 3)
The drug product resulting from a biopharmaceutical Particle shape (round versus fiber), particle optical properties
manufacturing process using SUT is subject to the particulate (transparency, color), and the particle surroundings (for example,
testing and specification requirements of the pharmacopoeial the transparency of the SUT) will likely strongly affect the particle
monographs. USP <1> Injectables (Ref. 20) requires that drug detection probability in a visual inspection of an SUT. The BPSA
products be 100% visually inspected for particulates, and that the considers particles smaller than 100 microns to be “sub-visible”.
drug product be “essentially free of visible particles”. Similar However, while USP, PDA and others consider particles greater
descriptions exist under the European and Japanese than 100 microns to be “visible” in the inspection of final drug
Pharmacopoeias using slightly different language: “practically products, this is likely not the case for the visual inspection of
free” of particulates (Ref. 21) and “free from readily detectable” most SUT. A study of a rigorous manual inspection process for
particles with unaided eyes. (Ref. 22) visible particles in SUT indicates that, in single-use bag
assemblies, the probability of detection for black particles first
The recent publications USP <790> Visible Particulates in
reaches >70 % at 500 microns. (Ref. 23) For tubing assemblies
Injections (Ref. 2) and associated guidance in USP <1790> Visual
(typically less transparent than bags), the probability of detection
Inspection of Injections (Ref. 3) recognize that a visual inspection
for black particles first reaches >70 % at 1000 microns, and clear
process is not 100% effective at detecting particles, and clarify
particles and fibers are even more difficult to detect. (Ref. 23) So
the “essentially free of visible particles” requirement by defining
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for most cases of SUT, the BPSA considers 500 microns to be the evaluate the risk of potential carry-over of visible particles by the
smallest particulate size likely detectable in a SUT visual pharmaceutical product through the different manufacturing
inspection. However, the SUT manufacturer should have a clear steps till the final container. To detect these particles, this
understanding of the typical levels of particles greater or equal to requires extraction of particles as described below.
100 microns. This is indeed key information for the end user to

Figure 9: Particle size classifications (not to scale)

500 microns
10 microns 25 microns 100 microns (smallest size visible in SUT)
0.13 microns (USP <788>) (USP <788>) (USP monographs)
0.3 microns
0.5-1 micron
sub-visible
visible

influenza
virus bacillus red
blood animal cell diameter of a
cell human hair
B. diminuta approximate
(Nisenet.org)
bacterium diameter of a
grain of salt

Part V: Particle Inspection & Quantification

For the manufacturer of SUT, the overall goal is to control and
reduce the level of particulate matter in or on SUT. According to
the ASME BPE 2019 standard, section III-13 Particulates (Ref. 24):
“Single-use components and assemblies
should be free of loose, non-embedded, and
solid particulates as seen by direct visual
observation without magnification.
Particulates greater than or equal to 100 µ
(m) are considered to be visible. The TAPPI
Size Estimation Chart may be used for Visual inspection and quantification of particulates is the
reference. Particulates less than 100 µ are responsibility of the SUT supplier. Methods for visual inspection
considered to be subvisible and should be and quantification of particulates must be fully qualified and
minimized.” implemented with qualified instrumentation and fully trained
personnel. The methods must be designed to yield reproducible
The significant challenges around developing visual inspection results which represent the actual level of particulate matter in
and quantification methods should not be underestimated. the SUT.
Particulate matter is small and usually dispersed at low levels Visual inspection and quantification methods for particulates are
over the generally large surface areas of SUT, and thus difficult to well described and standardized for final drug products, but no
visually detect or to collect together for microscopic analysis. It is standardized methods currently exist which are written
important to realize that particulate detection and quantification specifically for SUT characterization. Although it is currently
is a probabilistic challenge. Currently, the practically achievable common to claim that SUT meet USP <788> specifications, it is
probability of detection is less than 100%. important to understand that claiming the SUT meets USP <788>
specifications is not entirely sufficient for SUT characterization,
since USP <788> Particulate Matter in Injections (Ref. 5) describes

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two particulate measurement methods and particulate count testing is often called a “USP <788>” test, but in reality is a non-
specification specifically written for injections or infusions, not standardized adaption of a standard developed for final drug
SUT. USP <788> does not fully describe all the aspects of a product to SUT. (Ref. 25)
method for quantification of particulates in or on an SUT.
As discussed in detail below, if visual inspection of an SUT is
Procedures describing the liquid extraction of particulates are
complimented with liquid extraction plus particulate
missing, as discussed in more detail below.
measurement which only uses light obscuration (Method 1 in USP
However, the pharmacopoeial monographs for drug products do <788>), there is likely a significant gap in particulate detection
give important information on how to approach development capabilities (Figure 10). Published studies show that light
and qualification of robust methods for visual inspection, and obscuration, as typically practiced, does not reliably detect
describe methods to count and size particulates. For drug particles greater than 50 microns, well beneath the visible
products, two approaches to particulates detection and particle range. (Ref. 7) And a published study indicates that
measurement exist: 100% visual inspection (non-destructive) and significant probability of detection of particulates in a visual
destructive testing of a statistical sampling of the drug product. A inspection of an SUT starts at 500 microns in the best case
similar approach should be applied to SUT, since SUT are often scenario (black particles in translucent single-use bags). (Ref. 23)
difficult to visually inspect: bags are transparent to translucent, Hence, a combination of visual inspection along with destructive
but tubing more opaque and connectors are often totally opaque. sampling with light obscuration likely does not detect particulates
in the size range of 50 to 500 microns or larger. A significant “grey
The overall current status of the industry is that many SUT
zone” in particle detectability between light obscuration and
suppliers perform a non-destructive visual inspection of SUT, and
visual inspection exists, similar to that found in careful studies of
also destructive testing of SUT (during qualification of the SUT,
particulates detection in bottles. (Ref. 26)
and often a sampling during SUT production). The destructive

Figure 10: Approximate gap in particle detectability between visual inspection and USP <788> Method 1 (light obscuration)

VISIBLE
In
SUT

500 microns
METHOD
Approximate
2 gap between
Visible and
Method 1
detection
100 microns limitations

50 microns

METHOD
1
10 microns

0.1 micron

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Visual Inspection
Visual inspection of SUT, by both suppliers and end users, is an As described in USP <790> and <1790>, there is a probability of
important part of verifying SUT quality along the SUT lifecycle. A detection for particulates in a visual inspection process.
visual inspection process typically looks for the following non- Probability of detection can be maximized by optimization of the
particulate related potential defects: following parameters:
SUT bags • film creases/surface Lighting conditions intensity, angle
defects (reflected/transmitted),
• weld polarization
delamination/bubbles/misalignment
• holes/canals Background and contrast white, black (Ref. 21)
SUT • arrangement of
assemblies components (tubing, Presentation/manipulation vertical, horizontal
filters, connectors, etc.) as of SUT
described on the drawing
Scanning methodology top-bottom, left-right
• connection security
• holes/canals Inspection rate length of inspection, inspector
Packaging • overall integrity breaks

Relative to particulate-related defects, the visual inspection of Inspector training training sets with known
SUT must focus on detecting: defects
 Particulates (gels, foreign material) embedded in films and
Inspector fatigue ergonomics, inspector breaks
components
 Particulates on the interior (fluid contacting) surfaces of SUT
 Particulates on the exterior surfaces of SUT USP <1790> indicates that magnification methods potentially
increase inspector fatigue and may not necessarily improve
probability of detection. Inspection using unaided eyes remains
the basis against which any enhanced visualization system, or
semi- or fully-automated inspection system, is tested.
Visual inspection of SUT presents some significant challenges
relative to inspection of drug products. SUT are usually much
larger than ampules or syringes, and are usually assemblies of
components with various degrees of transparency. Large
assemblies, intricate assemblies with many components, and
assemblies with opaque materials will likely result in probabilities
of detection which are much lower than those found for typical
drug solutions for injection. (Ref. 3) Particulates on the exterior
surfaces or embedded in components are likely easier to detect
than particles on the interior (fluid-contacting) surfaces. Multiple
layers of packaging may hinder particle detection, thus SUT
Visual inspection using a light table
should be inspected before packaging steps.

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Figure 11: TAPPI size estimation chart

100 microns = 0.1 mm; 20000 microns2 = 0.02 mm2

(Reprinted with permission of TAPPI)

Table 1: Conversion of TAPPI areas into particle diameters extraction methods is useful in order to better understand
particulate matter risks in applications of SUT. Destructive testing
TAPPI Chart Circular Area Equivalent Diameter of SUT for particulates involves two steps: 1) liquid extraction,
(mm2) (microns) and 2) particulate counting and sizing. Such tests are typically
0.02 160 performed during qualification and also during routine
monitoring of the SUT manufacturing process, but not as a
0.05 252
pre/post-use test. The SUT cannot be used after liquid extraction
0.1 356 testing. When studying particulates, it is vital to understand and
0.2 504 control the extraction, counting, and characterization of the
particulates.
0.5 798
1 1128 Liquid extraction
In the application of SUT for the manufacture of
2 1596
biopharmaceuticals, particulate matter attached to the interior
5 2523 fluid contacting surfaces could potentially detach from surfaces
and contaminate process fluids, drug substance, or drug product,
Given the limitations of manual visual inspection, automated depending upon where in the manufacturing process chain the
visual inspection using camera systems certainly should be SUT is applied. The probability that a particulate will detach from
considered, especially for smaller and transparent SUT a surface depends upon the many physical-chemical forces which
components. USP <1790> indicates that automated visual control the level of adhesion of a particle to the surface of an SUT
inspection systems must be tested against manual visual material. Some types of particles may adhere to SUT surfaces
inspection to demonstrate effectiveness. In any case, automated more tenaciously than others. Some SUT surfaces may attract
visual inspection of SUT will be challenged by the same and retain/bind particulates more strongly than others.
constraints as the manual visual inspection of SUT: the large size
In addition, the probability that a particulate will detach from a
and complexity of SUT, and many SUT materials are translucent
surface depends upon the formulation
to opaque.
(proteins/stabilizers/excipients/buffers, etc.) of the process fluid,
Particle Quantification by Liquid Extraction drug substance or drug product, which surrounds the particulate
Since the probability of detection of particulates using visual and controls the “wettability” of the particle. Also important are
inspection methods may not be high in some cases, especially for flow conditions: faster flow during processing will likely apply
the interior (fluid-contacting) surfaces of opaque SUT assemblies, more shear to a particle attached to a surface, which likely
destructive testing using particulate quantification by liquid
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increases the probability for detachment of a particulate from an In essence, the liquid extraction step of an SUT is a washing or
SUT surface during biopharmaceutical manufacturing. flushing step using purified water or buffer. Care should be taken
to wet the surfaces of the entire fluid path, to consistently control
Similar considerations go into qualification of a destructive test
agitation intensity, and to minimize the amount of residual water
of an SUT using liquid extraction. The goal is to create a “worst
remaining in the SUT assembly after flushing. The total water
case scenario” and remove as many particulates as practically
volume applied should be adequate to achieve the extraction,
feasible from the SUT surface. In this way, as many of the
mobilization and suspension of the particles in the SUT test
particulates potentially detachable in real applications are
article. Care should be taken not to artificially create wear
quantified, which helps guide a risk assessment depending upon
particles from surfaces by using aggressive agitation which is
the criticality of the SUT application. It is important to note that
unrealistically intense.
the pharmacopoeial manuscripts do not provide guidance on
how to develop and qualify a liquid extraction protocol for SUT. Particle Counting and Sizing
Once a liquid extract (particulates extracted from the SUT and
An initiative to standardize the extraction of particulate matter
suspended in liquid) is generated, the pharmacopoeias (USP, EP,
from SUT was recently completed at ASTM International, and
JP) give useful guidance on how to properly count and size the
scheduled for publication in 2020 as ASTM E3230-20. The title of
particulates. However, it is important to understand that these
the proposed ASTM standard is “Standard Practice for the
methods have significant limitations when applied to SUT.
Extraction of Particulate Matter from the Surfaces of Single-Use
Components and Assemblies Designed for Use in Issues and limitations of applying USP <788> to SUT
Biopharmaceutical Processing.” Although USP <788> is often adapted for the analysis of
particulate in SUT, is important to understand that the USP <788>
The overall approach in the ASTM E3230-20 standard is a
standard is specifically written for drug products for injection or
“multiple extractions” approach, which is closely related to a long
infusion. When applying one of the USP <788> methods to SUT,
standing ISO standard for extraction of particulates from
two important aspects of the analysis of particulates in SUT are
automotive components. (Ref. 27) The same SUT component or
missing: the extraction protocol as noted above, and also how to
assembly is extracted multiple times during extraction method
report the particulate counts. For large volume parenterals in the
qualification. The qualification criterion described in this
USP <788>, the particle counts are given per milliliter of drug
standard practice is essentially the same as the “declining
product. For SUT, the particulate counts should be reported per
criterion” described in ISO 16232. In essence, this criterion
SUT component or assembly, per SUT fluid contact surface area,
requires that, during qualification, the chosen extraction
or per SUT nominal internal volume, noting it is important to
procedure must achieve an effectiveness of greater than 90%
separately report fluid contact surface area or nominal volume.
particle removal on a relative basis.
Reporting particulate counts per volume of extraction liquid
applied in the extraction protocol is not appropriate, since the
extraction liquid volume needed to maximize the efficiency of
extraction of particulates from SUT surfaces likely differs
depending upon SUT configuration.
Also note that USP <788> specifies maximum allowable levels of
particulates in small volume and large volume drug products.
These limits may not always be technically relevant for
determining SUT suitability for a chosen application. A “pass/fail”
assessment of using USP <788> limits is not appropriate for SUT.
Particle counts for SUT should be reported as noted above. If a
Qualification of an extraction protocol which is efficient in USP <788> large volume parenteral (LVP) limit is being applied,
removal of particulates involves consideration and optimization the data should be reported as particles per milliliter nominal SUT
of the following parameters: volume.
Type of extraction solvent most often purified water or Issues and limitations of the particle counting and sizing using
buffer, with or without methods described in USP <788>
surfactant USP <788> is entitled Particulate Matter in Injections and, as
Volume of liquid applied relative to the surface area or noted above, is a standard specifically written for drug products.
interior volume USP <788> is harmonized with EP 2.9.19. and JP 6.07 (Refs. 28 and
Type of agitation rinsing, shaking, sonication, etc. 29), and is closely related to USP <789> Particulate Matter in
Intensity of agitation shake frequency and number of Ophthalmic Solutions. (Ref. 30) USP <1788> Methods for
cycles Determination of Particulate Matter in Injections and Ophthalmic
Time, temperature (varies) Solutions (Ref. 31) is a guidance manuscript to assist in developing
Liquid flowrate (varies) a robust USP <788> or USP<789> method. Note that USP <787>

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Subvisible Particulate Matter in Therapeutic Protein Injections Method 1 and Method 2 as described in USP <788> and <1788>.
(Ref. 32), and the guidance document USP <1787> (Ref. 33) Table 2 highlights the differences, and lists the advantages and
primarily focus on the issue of measuring small inherent limitations associated with each method as assessed by the BPSA.
particulates (protein aggregates). With any particle counting and sizing method, accuracy may be
difficult to achieve, but precision (reproducibility) should be
Although USP <788> is written for drug products, the particle
achievable. (Ref. 7)
sizing and counting methods described in USP <788> and <1788>
are often used for the analysis of the liquid extract obtained from
SUT. USP <788> describes two methods for particulate counting
and sizing. In Method 1 Light Obscuration Particle Count Test, the
liquid extract flows through a light beam and particulates are
counted and sized. In Method 2 Microscopic Particle Count Test,
the liquid extract is filtered onto a membrane filter, the
membrane is placed under a microscope, and the particles on the
membrane are manually counted and classified by the analyst.
The particles on the membrane are visually compared to a
reference graticule (reticle) with 10 micron and 25 micron
diameter circles. .
It is important to note the significant differences between
Table 2: Comparison of methods described in USP <788>*

Method 1: Light Obscuration Method 2: Manual Membrane Microscopy

Indirect measurement using light blockage Direct visualization of particles on filter membrane using microscope
Particles detected by blockage of light beam Human operator eye is the detector
Particle size determined by amount of light Particle size determined by comparison with standardized graticule (reticle)
blockage detected by sensor, relative to amount of
light blockage by particle size calibration standard
Light blockage signal depends strongly on optical Visualization depends upon contrast between particle and membrane, lighting
and morphological properties of particle (e.g. conditions, microscope optical quality and operator training
transparency, shape) which often differ from
properties of particles used for calibration
No information on particle morphology and color, Direct visualization of particle morphology allows for accurate sizing, and
and often undersizes fibers and other irregularly facilitates particle identification based upon morphology. In addition, particles
shaped particles (e.g. glass fibers) are collected and available for application of more advanced methods of particle
identification (e.g. infrared, Raman or electron microscopy)
No filtration required: Filtration onto membrane required
Directly measure particles in liquid extract
Usually dilute: Usually more concentrated:
particles dispersed in liquid volume particles collected on membrane surface
Measures sub-visible particles, but typically does Will capture all solid sub-visible and visible particles larger than the pore size of
not detect “visible” particles (greater than around the filter membrane
50 to 100 microns). Larger particles in extraction
liquid tend to settle during measurement, and may
not be dosed into instrument and detected
Detection sensitivity may depend upon Does not measure water immiscible liquid droplets since droplets are not trapped
model/brand of instrument. Newer models often on membrane filter
show increased sensitivity, and may also measure
any water immiscible liquid droplets present (e.g.
from silicones)
Detects any air bubbles present Does not detect air bubbles
Less labor intensive and less subject to human Labor intensive and subject to human detection error, since each particle must be
error, since particles automatically counted and manually detected and sized by comparison with circles in graticule (reticle)
sized by instrument
* USP <788> (Ref. 5) and <1788> (Ref. 31)

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Automated Image Analysis of Particles Collected on Membrane Although size categories at 10 and 25 microns are stipulated in
Filters USP <788>, there is increasing interest in the biopharmaceutical
As noted in Table 2, the light obscuration method (Method 1) has industry in smaller particles. USP <787> and USP <1787> address
the advantage of ease of use (the instrument automatically measurement methods for particulates less than 10 microns,
counts and sizes particles in the liquid extract) which is one primarily focused on protein aggregates. ASTM E3060-16 (Ref.
reason why the light obscuration method currently finds 36) gives specific guidance on how to apply dynamic flow imaging
widespread use in the analysis of particulates extracted from methods to characterization of sub-visible particulates in
SUT. However, given the significant advantages of membrane biopharmaceuticals.
microscopy compared to light obscuration noted in Table 2, some
Critical Importance of Low Background Particle Counts
SUT suppliers have developed automated methods for the
Regardless of the extraction method and particle count/size
microscopic analysis of particles collected on filter membranes.
method chosen, a critically important aspect for a successful
Automated membrane microscopy eliminates the issues
particulates quantification is to achieve low background
associated with manual counting of particles on membranes
particulate levels. Background particulate levels are measured by
(labor intensive and subject to human error), relying on image
executing all steps of the extraction and count/size procedure,
analysis software to detect, count and size particulates.
but without the step of extracting the SUT test item. Care must
The automated counting and sizing of particles collected on a be taken to minimize particulate matter originating from foreign
filter membrane is not standardized for pharmaceutical sources other than the surfaces of the single-use components or
applications. Development and validation of such methods is assemblies of interest. Potential foreign sources include the
challenging, especially when the contrast between particle and extraction liquid, the delivery system for the extraction liquid, the
membrane is low. However, software and instrumentation for containers used to receive the liquid after extraction, particles
automated membrane microscopy has experienced rapid shed by the test operator (e.g. clothing fibers), and particles
development. The accuracy of counting and sizing has increased present in the general environment. It is recommended to filter
and the costs of automated microscopes have decreased. the extraction liquid, to carefully clean all extraction equipment,
Currently automated membrane microscopy is commonly to properly gown the test operator, and to perform the
applied in the analysis of particulates obtained from the procedures in a clean and environmentally controlled laboratory
extraction of automotive components. The ISO 16232 standard environment.
for automotive components and systems describes not only how
In the qualification of a particulate quantification by liquid
to develop and validate particulate extraction methods, but also
extraction method, the chosen approach to qualification of the
how to develop and validate particulate count/size methods
extraction method and the particle count/size method must be
using automated membrane microscopy. (Ref. 27) Automated
carefully documented, and all procedures required to execute
membrane microscopy shows significant promise for
the testing must be described in a standard operating procedure
measurement of both visible and sub-visible particles obtained
(SOP).
from pharmaceutical products, final containers, and from SUT.
However, a standard specific to measurement of particulates in BPSA Recommendations: Visual Inspection and Particle
pharmaceutical products using automated membrane Quantification
microscopy currently does not exist.
 Manual visual inspection of SUT should be optimized for
Other potentially useful methods for particle counting and particle detectability: lighting conditions, contrast, and
sizing inspector training plus inspection procedure (SOP) and
Given the limitations of the USP <788> methods, other methods accept/reject criteria.
might be of interest. For example, the ASTM F24 standard (Ref.  Automated visual inspection should be considered,
34) for small electronic components addresses counting particles especially for smaller and more transparent SUT
≥5, 25 and 100 microns found on the surface of electronic components.
components, or washed from a surface using sonication and
detergent. IEST-STD-CC1246E describes an often applied  Destructive testing (liquid extraction plus particle analysis)
standardized scheme for describing a surface cleanliness level should be carried out for SUT qualification, and periodically
with respect to particulates. (Ref. 35) on SUT components/assemblies during manufacturing, for
process capability and continuous improvement efforts.
Other potentially useful particulate counting and sizing methods  Liquid extraction of SUT should be optimized using a
include the Coulter counter and single particle optical sizing multiple extractions approach.
(SPOS). However, other light scattering techniques (diffraction,
static and dynamic light scattering and turbidity measures) only
 USP <788> is written for parenteral drug products and has
limited applicability to SUT: particle extraction methods are
provide particle size information, but no information on particle
not described, and the USP <788> particle limits only apply
counts.
to parenteral drug products. USP <788> tests can be useful,

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but are not sufficient for characterization of particulates in  Providing operator training;
single-use, specifically for visible particles.  Demanding cleanroom gowning;
 Particle counting and sizing using the light obscuration  Ensuring cleanroom maintenance and control; including
method (Method 1 in USP <788>) will only reliably measure proper housekeeping/cleaning practices
sub-visible particles: other methods are required for visible  Inspecting product, including up to 100% in-process
particles. surveillance and lot release testing;
 Automated membrane microscopy is a promising method  Recording performance trending for both the cleanroom
for counting/sizing visible particles, currently being applied operation and the manufactured SUT; and
by some SUT suppliers. BPSA recommends that this method  Documenting non-conformance, root cause analysis and
be seriously considered for determination of visible particle corrective/preventative actions.
counts in SUT.
Raw Components
 For SUT, particle levels should be only reported as particles
Monitoring and control of incoming materials, such as tubing,
per SUT volume, particles per SUT surface area, or particles
fittings, bag film, filters, connectors, tie wraps and packaging
per SUT component/assembly. Surface area and volume
components is critical to ensure proper particulate management.
should be reported in addition to particle level.
Simply put, the lower the particle load of incoming materials, the
greater 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. Alignment with the component supplier is vital.
. Discussions should be conducted with suppliers to ensure they
understand the criticality of their parts for the manufacturer of
the SUT.
Part VI: Control of SUT Manufacturing Assuring that the component suppliers are vigilant in their
Process practices is an absolute requirement. The purchase specification
of components should state the requirements. The component
Manufacturing of single-use assemblies normally occurs within a
supplier should have systems in place to ensure conformance to
cleanroom. Typically these are in an ISO 8, Class 100,000 or an
the particle requirements. Audit of the component supplier is
ISO 7, Class 10,000 environment. Classification should be
also recommended to ensure controls are in place and proper
measured by employing active air monitoring during the
handling and packaging methods are utilized to ensure low
manufacture of the single-use assembly and verifying the room
particle and bioburden load.
operation is within the applicable standard during that time
period. (Refs. 37 and 38) Although this space is very clean, it is An incoming inspection procedure for components may be
not certified as a “particle free” space. These standards use required at the SUT manufacturer to ensure material is not
measurements of airborne particulates in the size ranges 0.5 to 5 damaged during shipping and that it conforms to the
microns for the determination of cleanroom cleanliness, and so requirements listed in the purchase specification document.
do not measure particles in the sub-visible and visible size ranges.
Larger particles tend to settle quickly and stick to cleanroom Cleanroom Operation
surfaces. But ISO qualification of a cleanroom brings cleanliness A standard operating procedure should be in place for the flow of
rigor to cleanroom operations. The basic requirements to materials into the cleanroom, and for final assemblies leaving the
minimize particulates during the manufacturing of single-use cleanroom.
assemblies are as follows: It is recommended that incoming materials be multiple-bagged.
 Manufacturing processes designed to minimize the risk of This allows for the discarding of the outer bag, a source of
particle generation, introduction or inclusion into the contamination of cellulose fibers and other materials generated
finished assembly; during shipping. The inner bag(s) may then be sanitized with
 Maintaining proper preventative maintenance of alcohol or other disinfectant/cleaner and passed into the clean
manufacturing equipment; material storage area.
 Ensuring cleanliness of materials and people entering the Inspection and cleaning for particulates on raw materials
cleanroom; generally occurs during the kitting of materials prior to moving
 Controlling flow of materials and personnel within the these materials into the clean assembly area. During the
manufacturing environment; assembly process, work surfaces should be cleaned on a regular

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basis. Operators should be inspecting for particulate at the monitoring for particulates (viable and nonviable) in the
assembly operation. The final assembly is inspected, multiple- cleanroom is highly recommended to verify performance of the
bagged in the cleanroom, and transferred to the packaging area room to the cleanroom specifications. An established method for
outside the cleanroom for final packaging. periodic monitoring of the room under dynamic conditions
(during active manufacturing) is paramount to ensure
In addition to ensuring clean materials, procedures and training
contamination levels remain significantly below the accepted
should ensure a high level of operator cleanliness. A detailed
levels per the cleanroom classification.
procedure for an operator’s introduction into the cleanroom is
necessary to limit the transfer of particulate from outside the The monitoring process should encompass the entire facility and
cleanroom to within it. A dedicated gowning room at a minimum take into account how areas relate to each other throughout the
of ISO 8 is recommended. monitoring period. Additionally, to ensure proper operation of
the cleanroom, monitoring of temperature and humidity levels,
This room is set up with clean and dirty barriers to minimize
as well as air differential pressure between each classified area
contamination transfer. The SOP should outline the proper
and the non-classified area, must be performed.
conduct associated with entrance, operator conduct, and exit
from the cleanroom. Procedures for gowning and de-gowning An annual recertification for compliance to ISO 14644-1 is
should be clearly written, enforceable, and easily trained to, in required. (Ref. 37) The HEPA filters must be tested for airflow and
order to ensure maximum cleanliness of the operator. The SOP pressure drop and aerosol particle challenge. This ensures that
must outline the “do’s and don’ts” for cleanroom operation and the HEPA filters and HVAC system are functioning per design to
include a description of articles which are allowed in the ensure proper removal of contaminants and the appropriate
cleanroom and those that are restricted from the cleanroom. number of air changes in the room per hour.
Recommendations can include restriction of certain street
Cleanrooms should be cleaned on a periodic basis to assure
clothing and/or cosmetics worn by the operator prior to gowning.
proper operation and particle control. Routine operation and its
Reassignment to non-product contact activities is recommended
impact on particulate levels should be evaluated and cleaning
in the case of illness.
frequencies should be adjusted accordingly to assure cleanroom
Only approved cleanroom materials are utilized by the operators operation. Additional cleaning usually occurs in response to non-
including lint-free cloths, shoe covers, head bonnets, beard routine events: planned (maintenance) and unplanned
covers, and powder-free gloves. Training to the cleanroom SOP (deviations). Such events should be evaluated for their potential
must be regularly performed. Operators are encouraged to “self- impact on particulate levels and additional cleaning and/or rigor
police” each other in order operate at the highest level of of cleaning should be adjusted to suit the nature of each event.
cleanroom practice.
Figure 12 indicates the particle counts for an active cleanroom.
Cleanroom Performance Of importance is the separation between the maximum particle
Demonstrated process capability of the cleanroom's counts measured, the alert level, and the maximum particle
environmental monitoring is another important aspect of SUT counts (max) allowed. Action levels are typically set at 50% of the
manufacturing. Periodic (e.g. daily, weekly, monthly, quarterly) maximum particles allowed. The alert level is typically 25% of the
maximum level.

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Figure 12: ISO 7 cleanroom particle counts by week9: ISO 7 Clean Room Particle Counts by Week
Figure

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

 Tube cutting;
Manufacture of the SUT
 Final assembly of tubing, injection-molded fittings, bags,
Manufacturing equipment and processes used to make the SUT
filters and connectors;
play an important role in minimizing particles in the clean
manufacturing space. Equipment should be manufactured with  Leak testing; and
materials that are cleanroom compatible. This equipment should  Packaging
not shed particles or fibers, be easy to clean, and designed in a A rigorous final visual
manner to minimize areas where particles can build up. inspection process is required
Proper filtration of process air and venting of pneumatic valves prior to final packaging, with
utilized in automated equipment should be design considerations lighting and other conditions
and then properly maintained. clearly defined and
documented in a standard
Equipment placement should be done to avoid obstruction of operating procedure.
environmental airflow (intake and return) and include proper
preventative maintenance and operational controls to minimize Final product evaluation to
the risk of particle generation or accumulation. In some cases it agreed-to acceptance criteria
may advantageous to create a dedicated area(s) within the is recommended. If a
cleanroom with specially controlled airflows around equipment destructive test for particle
in order to prevent machinery-generated particulates from quantification is required, a
contaminating SUT. procedure for SUT extraction
and particulate measurement
Visual checking is recommended before, during, and after all must be clearly defined and
process steps. These include the following: documented in a standard
 Bag manufacturing operating procedure. If the
o Film cutting; assembly fails the release test, Automated packaging system
o Port bonding; and material should be using aspiration to remove
o Film sealing quarantined, and an any remaining loose particles
investigation launched to
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assess the reasons for the out-of-specification test result. blow air used to clean parts, wipe down with IPA instead of water,
Corrective actions may be put in place after the investigation to drying rates vs. humidity of cleanroom.
limit failures in the future SUT Supplier Mitigation.
Change Management
SUT suppliers should have quality programs in place to help drive Changes in the specifications and/or testing methods should be
continuous improvements related to particle mitigation communicated to the suppliers and end users via the Change
strategies. There are particle mitigation processes/equipment Notification Practices for SUT per the BPSA/BPOG documents
that SUT suppliers can implement in their SUT manufacturing entitled An Industry Proposal for Change Notification Practices
process steps, but due to types of components used in SUT (i.e., for Single-Use Biomanufacturing Systems and A Guide to the
silicone tubing, small plastic couplers, etc.) the effectiveness of Classification of Changes to Single-Use Biomanufacturing
these mitigation steps can vary greatly or introduce other Systems).
undesirable risks. Suppliers need to balance what is realistic vs.
the goal of zero particles. For example, an air blow down process Product Specification
may help reduce sub-visible and visible particle levels in The product specification and particulate level specification
components such as rigid connectors, but will likely not be as would be detailed in a user requirement document such as the
effective with silicone tubing due to ID of the tubing, length of the BPSA User Requirements Template.
tubing, and tackiness. A liquid wash down of components less
responsive to an air blow down will lead to bioburden, endotoxin
and extractables risks that complicate the SUT manufacturing
Part VII: Control of Biopharmaceutical
process and require the establishment of validated drying Manufacturing
procedures. Two phases in the lifecycle of an SUT need to be considered and
An alternative that is helpful for particulate removal is the use of integrated when analyzing particle generation and control. The
an aspirator. The purpose of the aspirator is to reduce static on first phase includes the supplier’s sources, manufacturing and
the parts, using ionized air, prior to packaging operations. This assembly, packaging, and shipping. The second phase includes
process is meant to reduce debris that may be clinging to the end user receiving, handling, and use of the SUT. Particles are
parts because of static electricity. This process is effective in introduced to, and generated within, the SUT in both phases.
removing process and non-process related particles. Table 3 lists some of the ways that end users can introduce
particles to the SUT.
Other options are air blow with deionized air, mitigating the
electrostatic charge of the single SUT, venting/exhausting air

Table 3: 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 (including final drug packaging containers) can interact with
material ingredients/product components of a protein solution to form precipitates. These can be further exacerbated by process
conditions and/or type of single-use component
Manufacturing activities Connecting and disconnecting assemblies
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)
Manufacturing environment Open system applications of single-use
Personnel Handling of SUT assembly or part(s)

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11. Match peristaltic pump tubing type and dimensions to pump

Processing Considerations heads, process duration, and process fluids. Do not exceed
Evaluations of particle risk in end user manufacturing should
anticipated tubing life.
include placebo runs that use worst-case process conditions.
12. Minimize surfaces that can rub together during shipping,
Worst-case conditions are those that are reasonably expected to
storage, or use.
occur and that present the greatest potential for particle
generation.
Part VIII: Deviation Response/Mitigation
Parameters to consider are:
 Mixing speed;
Plans
 Number of connections made during the process; When a visible particle(s) is observed in an SUT, whether during
 Storage times and temperatures; manufacturing or at the end user’s facility, the response depends
 Line clamping or valve use; on a number of factors. Perhaps the most important
 Pumping/spallation; consideration is whether or not the particles are a deviation from
 Welding or sealing tubing; the quality standard or specification.
 Rinsing/flushing/washing steps*; and Purchasing specifications and inspection criteria should make the
 General handling practices particle requirements clear. Number, size, and type of any
allowed particles on or in an SUT should be agreed to between
It is recommended to run the process in the intended production
supplier and end user via a clearly written specification. The
facility or, at minimum, under the process and product
specification should take into consideration the process
manufacturing conditions that are planned.
capabilities of the supplier as well as the quality requirements of
(*At times, some biopharm manufacturers flush the lines and containers the end user. It is important to identify gaps between supplier
prior to use with the intent of removing any particulates or may be built capabilities and end user expectations.
into the filtration manifold. This can add complexity to the system (i.e.
dilution of biopharm ingredients). Scientific justification or justifiable The next sections follow a single-use item through its lifecycle
outcome of a risk assessment should be in place.) 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
Best Practices for Handling Single-Use described.
Components A good particle investigation requires strong cooperation
Although all of the items below may not directly affect particles, between end users and suppliers. Both parties need to avoid
these are recommended practices for SUT in general: defensive or accusatory attitudes that do not facilitate the open
1. Cover sharp parts. Do not remove supplier’s protective communication and clear thinking needed to resolve problems
coverings until necessary. quickly.
2. During storage, bags should be contained in a hard-shelled
container or, at minimum, covered with a sealed outer bag. When in the SUT Lifecycle is the Particle
Lines should be secured as appropriate, especially when
freezing.
Observed?
3. Flush the systems, especially those that contain filters or fiber- The stage in the SUT lifecycle in which a particle is found affects
shedding components, where possible. the type and intensity of investigation activities. Consider the
4. Avoid over-processing: over-mixing or over-handling of following when preparing an investigation plan:
components/assembly. Particle found during SUT subcomponent manufacturing and
5. Avoid pulling, flattening, rubbing, squeezing, flexing, or assembly:
twisting of components/assembly. Investigate as a cleanroom management deviation.
6. Optimize the welding and sealing conditions to avoid
“flashing” or inadequate welds. Particle found during QC inspection prior to packaging by the
7. Keep product fluid contact path as short and with as few supplier:
components as possible. More extensive investigation is required: characterize the
8. Do not lift items by their tubing connections. particle and establish a root cause; document for statistical
9. Minimize the stress on tubing junctions. Avoid sharp bend process control (SPC) purposes; and evaluate “related lots”
radii. (those in the facility at the same or adjacent processing times and
10. Do not allow sharp objects to be used in the same area as lots prepared with common components).
single-use components. Particle found after packaging and before shipment to end
user:
More extensive investigation is required including a check on the

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QC processes and their effectiveness. Particle characterization is user and requires a root cause investigation. A commercial
needed and a root cause should be pursued aggressively. complaint should be filed if this is out of specification. The SUT
Document this for SPC purposes. 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
Particle found at end user receiving:
should be assessed.
This is potentially not a quality system problem for the end user.
A commercial complaint will be filed, provided that the part in Particles observed on the exterior of SUT or embedded
question is out of specification. An investigation should be particles:
performed by the supplier for a post-packaging event. Shipping The investigation intensity depends on the nature of the
configuration and method should be assessed as potential particles, SUT specifications, and the stage of end user processing
contributing root causes. as described above. The supplier should be notified and a
complaint filed as appropriate so that the observations can be
Particle found at point-of-use (before use):
included in trending analyses.
This is a quality system problem for the end user and needs to be
assessed for impact on the cleanroom environment. Root cause Particles observed within the process/product contact surface
investigation needs to consider both end user and supplier as (wetted path) or in areas that connect with the wetted path
potential sources of the particles. A commercial complaint will be such as vent lines and unused tubing legs:
filed to obtain supplier assistance and the event counts against This is a serious quality system problem as discussed above. If the
supplier’s quality performance score provided that the supplied SUT is being used, quick decisions are needed about continuation
part is out of specification. An investigation should be performed of processing and quarantines of end user product and SUT. Root
by the supplier as for a post-packaging event. cause determination needs to consider both the end user and
supplier as potential sources of the particles. A commercial
Particle found at point-of-use (during use):
complaint will be filed to obtain supplier assistance. An
This is a quality system problem and quick decisions are needed
investigation should be performed by the supplier as for a post-
regarding continuation of processing and quarantines of the end
packaging event if it is found to be outside of specification.
user’s product and SUT. Root cause determination needs to
consider both end user and supplier as potential sources of the
particles. A commercial complaint will be filed to obtain supplier Particle Investigation Steps
assistance. An investigation should be performed by the supplier Given the general considerations about when and where a visible
as for a post-packaging event. particle is observed, the following steps are typically taken for
investigations involving suppliers, end users, or both. Some of the
Particle found after processing is complete:
steps are unique to end users and their relationships with
Actions taken are the same as for those at point-of-use. End user
patients and regulatory agencies. Risk posed by particles must
decision-making about quarantines will be urgent.
first be considered in terms of patient risk. Roles and
responsibilities are dependent on the specific situation.
Where is the Particle Observed – Step 1. Detect a particle (or particles) during SUT
On or In the SUT? manufacturing, quality inspection, or during end user
Particles that contact the end user’s process/product stream are operations.
significantly more serious than those on the exterior of the SUT Step 2. Report the finding via formal quality system
or packaging materials. While process/product contact surface procedures (deviation report, event notification, etc.).
(wetted path) cleanliness has top priority, the mode of use of SUT Suppliers and end users should alert each other as
may warrant concern about particles outside the wetted path. required by quality agreements or other agreements.
Reporting should occur even if the SUT is within
For example, an SUT used in non-closed system fashion or within
specification so that trending analyses can be
a Grade A environment needs to be as close to particle-free as
performed.
possible. Unusual requirements like this need to be discussed
with the supplier during the design process. Step 3. Hold the lot (if possible) or continue at risk. Immediate
discard may be required (see Step 9). The end user’s
A general approach for reacting to particles based on their product batch is placed under administrative
observed location is as follows: quarantine (and possibly physical quarantine).
Particles observed on the outer package (bag): Step 4. Quarantine related SUT parts as a precaution until a
These are usually not handled as an end user quality system root cause investigation provides more information.
problem. Step 5. Capture and characterize the particle(s). Count if
Particles observed on the inner package (bag that was over- there are too many to characterize each one.
packed in a clean environment): Step 6. Compare the particles to a catalog of previously found
This is a quality system problem for both the supplier and end particles.

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Step 7. Use characterization data and catalog matches to Step 12. Review investigation and quality decisions with
determine the source of the particles and a root cause regulatory body representatives during inspections as
for their appearance in the SUT. Materials of required. Consider this activity when preparing the
construction of the SUT, and the end user’s other investigation report and supporting documentation.
production system parts, are often the source of Step 13. Identify and complete corrective and preventative
particulates in SUT. Cleanroom supplies and clothing actions that may involve the disposition of the batch,
are also common sources of particles discovered in disposition of single-use parts and further steps to
SUT. Look for obvious sources before chasing exotic prevent repeat investigations. Actions may include
hypotheses. Analytical data is often tenuous for small changes to either the end user’s manufacturing
particles. Partner with the analytical services provider process and/or changes to single-use supplier
to understand both what the compositional manufacturing, packaging, or sourcing.
assignment is as well as what plausible alternate
assignments are. Knowledge of SUT materials
provided by the supplier and the end user component
engineer will help the analyst to run appropriate tests Part IX: Summary & Conclusion
and assist in the interpretation. Information about As an important part of the chain of stakeholders in the
suppliers who are located earlier in the supply chain manufacture of biopharmaceuticals, single-use suppliers must
may also be needed to identify the source of particles. continuously focus on improving SUT manufacturing in order to
Step 8. Assess the impact on other SUT parts. Add minimize particulate levels in SUT, especially for critical
quarantines and, if necessary, remove quarantines or applications of SUT after final filters. Only a holistic life-cycle
make rejection/discard decisions. approach to the issue of particulates in SUT will satisfy the
requirements of SUT end users, regulatory agencies, and
Step 9. Assess the end user product batch impact and make
ultimately satisfy the safety needs of the patients of
“disposition” recommendations. Product quality
biopharmaceutical therapies.
decisions depend heavily on where the particles are
found, the ability of the investigation team to prove There are four primary areas that must be managed in order to
that similar particles are not present in other ensure robust control of particulates in single-use systems:
locations, and on the chemical identity of the
1. Cleanliness of the incoming materials;
particles. Positive quality decisions (those that allow
2. Cleanliness of the manufacturing steps and assembly
further use or release of a batch) are not possible
processes;
without a convincing root cause investigation. In the
3. Cleanliness of the operators and associated gowning;
absence of sufficient detail, end user product should
and
be discarded.
4. Cleanroom facility and equipment maintenance,
Step 10. Assess the impact on previously manufactured end monitoring, and controls.
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 the
product, attention to these four areas will ensure the end user of
Step 11. Prepare an investigation report and obtain approvals
minimum visible particulates in the SUT product.
for root cause analysis and product disposition
decisions. Figure 13 illustrates how effective controls can reduce the
number of particulates, specifically those from outside sources.
Less particulate is better!

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Figure 13: Use of controls over time to reduce particulates

Environment
Environment
People
controls People controls controls

SUT SUT
Components Components

“Initial” “Final” “Target” “Perfect World”

Particulate Profile Particulate Profile Particulate Profile Particulate Profile

TIME

consequently, decrease the risk of harm to patients from

Recommendations particulate contamination.
Control of particles in SUT manufacturing should use a risk-based
2. The BPSA recommends that suppliers of SUT develop a
approach and will require diligence from all suppliers in the
strong understanding of the particulates (sizes, levels,
supply chain, as well as during the assembly of the SUT. Particle
types, and sources) which might be present within their
control is everyone’s responsibility, including the end user.
SUT. Concepts for exchange of information on particulates
Because manufacturing and use of systems are not performed in
between suppliers and end users should be developed and
a particle-free environment, systems should be implemented to
implemented.
ensure cleanliness and monitor performance.
3. The challenges in developing rigorous visual inspection and
An ISO 8 or better environment is recommended for SUT particle quantification methods for SUT should not be
assembly and component manufacturing. Many assemblers now underestimated! The goal of visual inspection and
manufacture in ISO 7 and ISO 5 environments to better control quantification methods is to maximize the probability of
their manufacturing environments and thus lower particulate detection.
risk. In addition, extra diligence, and steps such as pre-cleaning of 4. The goal of a chosen particulate extraction protocol for SUT
parts by the component supplier, may be required to ensure is to maximize the probability that particulates are
parts meet the strict incoming needs of the SUT manufacturers. extracted in a practical, consistent and controlled way,
which gives useful information for realistic assessments.
Particle control for all particles, both visible and sub-visible, in
5. USP <788> Method 1 and Method 2 both have significant
single-use assemblies is critical to quality. Quality must be built-
limitations, but Method 2 is preferred. While Method 1 is
in by ensuring proper systems are in place to minimize
quicker and easier to apply, light obscuration may not
particulates in the manufacturing of SUT. Relying on visual
detect particles in the visible size range. Method 2 is labor
inspection and final lot release testing does not ensure that a
intensive and subject to human error, but does detect
perfect assembly is supplied to the end user.
particles in the visible size range.
Instituting the systems and training outlined in this document will 6. Automated membrane microscopy is a promising method
assist SUT suppliers (and their suppliers) in meeting and for counting/sizing visible particles, currently being applied
exceeding end user expectations for control of particulate by some SUT suppliers. BPSA recommends that this method
contamination. be seriously considered for determination of visible particle
counts in SUT.
1. When it comes to particulates in biopharmaceutical
7. Automated visual inspection should be considered,
processing, fewer is better! Both SUT suppliers and end
especially for smaller and more transparent SUT
users are taking many steps to decrease the levels of
components.
particulates in their products. This is in order to minimize
8. The BPSA recommends that suppliers of SUT implement
the risk of contamination of drug products and,
manufacturing processes and environments that reduce the
risk of particle contamination.

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Continuous Improvement – Particle Reduction Part X: BPSA-Recommended Next Steps

and Simplification of Investigations As stated in the Introduction, this document captures the current
SUT suppliers and end users need to cooperate to control state of the industry with regard to SUT and particulates.
particles and make further reductions. Control and improvement Through the writing of this document, the group discussed at
efforts should consider the full lifecycle of the SUT, from length many items that are not fully addressed here and which
manufacturing supply chains to end user deployment in they feel need further study. These items include:
biopharmaceutical production, rather than emphasizing a
supplier vs. end user perspective. 1. USP <788> is not adequate for characterization of SUT. The
soon-to-be published ASTM standard E3230-20 will describe
While these practices will help with any supplier/end user procedure for optimizing particle extraction from SUT. Since
relationship, a view across the full set of suppliers and end users particle counting/sizing using USP <788> Method 1 does not
would be powerful. The following best practices are capture visible particles, methods specific to particle
recommended: counting/sizing particles from SUT need standardization.
 Continuous monitoring and trend analysis: Trending 2. Automated detection methods for visual inspection and
analyses for cleanroom performance and for particle membrane particle counting have promise, but will benefit
observations made throughout the SUT lifecycle should from increased application and standardization between
be performed; methods.
 Continuous characterization: Captured particles should 3. Application-specific requirements need to be better defined.
be identified; These include final bulk product vaccines and cell therapies
 Continuous cataloging: Particle identifications should be that require lower levels of particles than other routine
added to an organized and shareable catalog wherever processes more typically subject to final sterilizing filtration
possible; to control particulates in filled products.
 Continuous improvement activities should be evaluated 4. A format for confidential sharing and assessment of industry
by trending analyses; and progress in SUT cleanliness for particulates is needed.
 Periodic sampling per lot, per week. 5. Particulate generation studies using simulated end user
processes should be conducted to verify best handling
These steps will help the industry to demonstrate that we practices.
understand the particles that may be in SUT, and that the systems 6. Supplier/End User Quality Agreements of acceptance criteria
to manage particles are in control. We must apply continuous for the SUTs should be established. These should utilize
improvements, track results, and show that the particle control many of the principles mentioned in this document and
process is improving. 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.
7. A format for exchanging information between SUT suppliers
and end users regarding “technically difficult to avoid
particulates” should be established. As an example,
excipients suppliers have a guidance document on the
“Technically Unavoidable Particles Profile Guide”. (Ref. 39)
8. BPSA should coordinate with USP on forthcoming <667>
document.
Figure 14 provides an example of how suppliers and end users
might discuss and document their agreement. In the figure, the
dotted line represents the agreed-to acceptance criteria for a
specific application. This may vary for each SUT and each
application.

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Figure 14: Example of end user/supplier agreement for particulate acceptance criteria

MATERIALS OF
ACCEPTABLE NONE PACKAGING ACCEPTABLE
USP <788> CONSTRUCTION
(less than LVP limit)

USP CLASS VI OUTSIDE

MATERIALS OF
CONSTRUCTION
EMBEDDED
USP <788> ENVIRONMENT
(meets LVP limit)

FOREIGN EXTRINSIC
USP <788>
(greater than LVP limit) VIABLE

HAIR/BUG TOXIC PROCESS/PRODUCT

UNACCEPTABLE VIABLE CONTACT UNACCEPTABLE

VISIBLE SUB-VISIBLE CHEMICAL LOCATION

(≥ 100 microns) (< 100 microns) MAKE-UP

Indicates the agreement level for each application

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Part XI: Terms & Definitions

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 (protein) “Any virus, therapeutic serum, toxin, antitoxin, or analogous product applicable to the prevention,
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)

Clean build The manufacture of the SUS in controlled conditions, including ISO-rated cleanroom and proper
environmental and procedural controls to reduce the levels of particulate and potential
contaminants. Utilization of materials which are low particulate in both the components and
environment are encouraged.

Cleanroom “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)

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
ASME BPE)

Endogenous Endogenous particulates are those derived from constituents of the pharmaceutical process or
product. These particulates are a result of the drug 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 (formulation) Existing in the material as a permanent, essential, or characteristic attribute.

Intrinsic (material of The process contact materials of the SUT. Intrinsic particles are part of the process/product like
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.

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

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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)

Count/Quantity 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 typical
(SUT) [also known as systems are made up of bag chambers, connectors, tubing, and filter capsules.” (per BPSA).
single-use systems (SUS)]

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>)

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Part XII: References

1. PDA Technical Report No. 79, Particulate Matter Control in Difficult to Inspect Parenterals, Parenteral Drug Association, 2018
2. USP <790> Visible Particulates in Injections, United States Pharmacopeial Convention, 2017
3. USP <1790> Visual Inspection of Injections, United States Pharmacopeial Convention, 2018
4. S. Langille, PDA Journal of Pharmaceutical Science and Technology, 67, 186, 2013
5. USP <788> Particulate Matter in Injections, United States Pharmacopeial Convention, 2012
6. Quality Risk Management, International Conference on Harmonization, ICH Q9, 2005
7. T. A. Barber, Control of Particulate Contamination in Healthcare Manufacturing, CRC Press, 1999, ISBN: 1574910728
8. S. Bukofzer et al., PDA Journal of Pharmaceutical Science and Technology, 69, 123, 2015
9. D. Clarke et al., Cytotherapy, 18, 1063, 2016
10. A. Yun, et al., Journal of Bacteriology and Virology, 40, 11, 2010
11. J. S. Bee et al., Journal of Pharmaceutical Sciences, 98, 3218, 2009
12. A. Tyagi et al., Journal of Pharmaceutical Sciences, 98, 94, 2009
13. ASTM D883-20, Standard Terminology Relating to Plastics, 2020
14. K. Wormuth, et al, Embedded Particles in Single-Use Bags, BioProcess International, 17(7)E1, July 2019, p. 1
15. B. Gikanga, et al. PDA Journal of Pharmaceutical Science and Technology, 71, 172-188, 2017
16. Saller et al. Journal of Pharmaceutical Sciences, 104, 1440, 2015
17. PDA Technical Report No. 54, Implementation of Quality Risk Management For Pharmaceutical and Biotechnology Manufacturing
Operations, Parenteral Drug Association, 2012
18. R. A. Carlton, Pharmaceutical Microscopy, Springer 2011
19. K. Wormuth, et al, Visible Particulate Matter in Single-Use Bags, BioProcess International, 17(4), April 2019, p. 50
20. USP <1> Injections. U.S. Pharmacopeia National Formulary, 2013. USP <36>. pp. 33-37.
21. EP 2.9.20. Particulate Contamination: Visible Particles. 2008; European Pharmacopoeia 6.0; 01/2008; pg. 323
22. JP 6.06 Foreign Insoluble Matter Test for Injections. Japanese Pharmacopoeia, 17th Edition
23. K. Wormuth, Reduction of Risks from Particulate Matter in Single-Use Systems, Presentation to PDA Europe BioManufacturing, Sept 4,
2019
24. ASME BPE 2019, American Society of Mechanical Engineers, Bioprocess Equipment Standard 2019
25. J. Vogel and K. Wormuth, Particulate Contamination in Single-Use Systems, BioProcess International, 2017
26. M. Berdovich, https://www.interphex.com/Education/Presentation-Archive/2016/Using-Optical-Microscopy/
27. ISO 16232:2018 Road vehicles – Cleanliness of components and systems, International Standards Organization, 2018
28. EP 2.9.19. Particulate Contamination: Sub-Visible Particles. European Pharmacopoeia 6.0; 04/2011; pg. 321-323.
29. JP 6.07 Insoluble Particulate Matter Test for Injections. Japanese Pharmacopoeia, 17th Edition
30. USP <789> Particulate Matter in Ophthalmic Solutions, United States Pharmacopeial Convention, 2012
31. USP <1788> Methods for Determination of Particulate Matter in Injections and Ophthalmic Solutions, United States Pharmacopeial
Convention, 2012
32. USP <787> Subvisible Particulate Matter in Therapeutic Protein Injections, United States Pharmacopeial Convention, 2014
33. USP <1787> Measurement of Subvisible Particulate Matter in Therapeutic Protein Injections, United States Pharmacopeial Convention,
2015
34. ASTM F24-09 Standard Test Method for Measuring and Counting Particulate Contamination on Surfaces, ASTM International, 2015
35. IEST-STD-CC1246E, Product Cleanliness Levels – Applications, Requirements and Determination, Institute of Environmental Science and
Technologies, 2013
36. ASTM E3060-16, Standard Guide for Subvisible Particle Measurement in Biopharmaceutical Manufacturing Using Dynamic (Flow)
Imaging Microscopy, ASTM International, 2016
37. ISO 14644:2015, Cleanrooms and Associated Controlled Environments, International Standards Organization, 2015
38. FED-STD-209E, Airborne Particulate Cleanliness Classes in Cleanrooms and Cleanzones (Superseded in 2001 by ISO 14644)
39. Technically Unavoidable Particles Profile Guide, The International Pharmaceuticals Excipients Council, 2015

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APPENDIX A: BPSA User Requirement Template Information Relative to Particulates in SUT

This table contains partial information from the BPSA’s User Requirement Template. End users are instructed to use this table to
provide detailed, relevant, non-confidential information to their suppliers about their functional, qualification, and quality
requirements relative to particulates in the single-use system. The full BPSA User Requirements Template can be found HERE.

ID REQUIREMENT DESCRIPTION SUPPLIER RESPONSE

Endotoxin:
18 ⃞ Aligned to USP <85> or Ph. Eur. 2.6.14
⃞ Other: _____________________________________
Visible particulates:
⃞ Aligned to USP <790>, Ph. Eur. 2.9.20 or JP 6.06
⃞ Other: _____________________________________
19 Sub-visible particulates:
⃞ Aligned to USP <788>, Eur. 2.9.19, or JP 6.07
⃞ Aligned to USP <789>
⃞ Other: _____________________________________
SUS manufacturing/assembly environment classification:
Requirement for the SUS to be manufactured in an environment
as indicated, or in a more tightly controlled environment
⃞ ISO Class 5
⃞ ISO Class 6
30
⃞ ISO Class 7
⃞ ISO Class 8
⃞ Controlled non-classified space
⃞ Other: _____________________________________

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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.

This document is not intended to, nor should it be used to support a cause of action, create a presumption of a breach of legal duty,
or form a basis for civil liability. Nothing expressed or implied in this informational document is intended, or shall be construed, to
confer upon or give any person or entity any rights or remedies under or by reason of this informational document.
Determination of whether and/or how to use all or any portion of this document is to be made in your sole and absolute discretion.
Use of this document is voluntary.

BPSA shall not be responsible or liable for any inaccuracies in the document or the information presented. All warranties express or
implied are disclaimed and waived.

Manufacturers, suppliers and end users should consult with their own legal and technical advisors relative to their SUT use and
participation. No part of this document constitutes legal advice.

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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