Particulate Matter:

USP Requirements and Particle Identification

Bryan Wickson 2019-04-18

 Background

• I have worked at Exova for 19 years

• First 8 in the field of polymers and medical
device development
• L a s t 11 y e a r t e s t i n g o f m a t e r i a l s a n d
pharmaceuticals
• Mainly spectroscopy and physical
characterization

• Exova has 3 GMP labs:

• To r o n t o
• Delaware
• Santa Fe Springs
 Background

• Formulation development
• Clinical trials manufacturing
• Compendial testing, development and
validation
• E&L plus stability studies
 Agenda

Review of USP Requirements

Approaches to Identification of
Particulate

Case Studies
 Biological Response

Particles in the vascular system:

• blockages
• emboli
• accumulation/chronic damage to organs

Extravascular particulate:

• immune responses
• eye/tissue damage
 Biological Response

Real Life Examples:

• Glass fragments in inhalers

• Aluminum slivers in ophthalmic

eye drops

• Rubber o-ring pieces in

IV solutions
 Background

Particulate matter testing of pharmaceutical

parenteral solutions is governed by:

• USP<1>Injections and Implanted Drug Products

• USP<1790><790>Visual Inspection of Injections

• USP<1787><787>Sub-visible Particulate Matter

in Therapeutic Protein Injections

• USP<1788>Particulate Matter in Injections and

Ophthalmic Solutions
• USP<788>Particulate Matter in Injections

• USP<789>Particulate Matter in Ophthalmics

 Background: USP

Particulate matter can be defined by size:

• Visible particles: ≥ 100 μm

• Sub-visible particles: < 100 μm

And by source:

• Inherent
• Intrinsic
• Extrinsic
 USP<1>

“Should be prepared in a manner designed to

exclude particulate matter”

“Each final container of all parenteral

preparations should be inspected to the extent
possible for the presence of visible particles”

“Every container in which the contents show

evidence of visible particulates must be rejected”
 USP<790> Visible Particulate

• Specific lighting with different backgrounds

• Operators are trained and qualified

Black
White
Background
Background
USP<787><788><789> Sub-visible Particulate

• No visible particles are allowed

• USP contains limits for sub-visible particles

• Method I: Light Obscuration Particle Count

• Method II: Microscopic Particle Count

 Method 1: Light Obscuration

Laser Diode

The technique involves

directing a laser through a
dilute solution

Flow

Instrument counts and

sizes particles Shadow

Detector
 Method 1: Light Obscuration

Pros

• 1-600 microns
• Fast and repeatable
• Recommended by USP

Cons

• Provides “average” particle size

• Instrument accuracy
• Bubbles

NOTE: The method should be verified for the

specific product and any unusual sample
preparation
 Method 2: Microscopic Particle Count

Sample is filtered

Particles are sized/counted by using light

microscopy:

• Manual counting

• Automated stage/Image analysis

(good for high volume QC testing)
 Method 2: Microscopic Particle Count

Pros

• Potentially more accurate that Method 1

• Not impacted by artefacts such as bubbles

• Appearance of particle

Cons

• More time consuming

• Image analysis requires

validation/verification

• Open to interpretation…
 Next Steps

An investigation is required to determine the

nature and source of the particulate

Investigative analysis:

• Va l i d a t e d m e t h o d s a r e n o t r e q u i r e d
 Contaminant Analysis

Identification of particulate

The particulate may be visible or sub-visible

Identification is key:

• Improve processes
• Maintain regulatory compliance
 Contaminant Analysis

How are the particles found?

• Incoming inspection of raw materials

• Operators
• QC inspections of final product
• Pharmacists/nurses
• Patients
 Contaminant Analysis

Impact of contaminant analysis

Potential outcome:

• Normal release of the final product

• Process modification
• Substitution of process equipment
• Rejection of a Lot of raw material
• Rejection of a final product
• Product recall
 Sample Preparation

• Filtration (USP <788> Method 2)

• Centrifugation

• Manual removal with light

microscope and scalpel

• Solvent
extractions/rinsing
Sample Preparation
 A n a l y t i c a l To o l s

Te c h n i q u e s m u s t b e s u i t a b l e f o r a n a l y z i n g m i c r o n
sized particles typically weighing less than a few
micrograms

These include:

• Light microscopy

• SEM and SEM/EDS

• Micro-FTIR
spectroscopy
 Light Microscopy

•Colour and size of a contaminant

•Physical nature of the contaminant:

•Gel-like
•Solid (crystal, amorphous or fiber)

•Heterogeneity of the contaminant

•Chemical nature of the contaminant:

•Organic material
•Polymeric material
•Inorganic (e.g. salt or metal)

•Recommendations for analysis

starting point
 FTIR Spectroscopy

• FT IR is similar to UV-Vis
• Light is passed through a sample and the
absorption of light is measured.
• U V- Vi s s e n d s 1 wa v e l e n g t h a t a t i m e
• FTIR simultaneously
measures a variety of Stationary Mirror
wavelengths

Light
Source
Beam
Splitter Moving Mirror

Sample

Detector
 FTIR Spectroscopy

• The radiation causes the sample to vibrate and

absorb energy
• Different molecular bonds absorb at different
characteristic frequencies
• Different bonds can vibrate in multiple different
manners (bending, rocking, stretching) resulting
in multiple absorbances
• The IR signal can transit through a sample or
g r a z e ( r e f l e c t ) o f f a s a m p l e s u r f a c e ( AT R
technique)
 FTIR Spectroscopy

• Here is an example Polyethylene spectrum

-(-CH2-CH2-CH2-)n-

C-H stretching Scissoring

of H’s

Rocking
of H’s
 FTIR Spectroscopy

Can be used to analyze 10-20 µm sized particles

The spectrum can be compared reference spectra

FTIR is best suited for polymers and organic

materials

Mixtures of materials can be identified by using

spectral subtractions
 SEM/EDS
Electron Beam

• SEMs are traditionally Backscattered

used to obtain morphology Electron
of conductive materials Detector
(i.e. metals)
• This is performed by
collecting emitted
secondary electrons
emitted from a surface e-
e-
• Collection of emitted X-
rays allows for the
determination of the
source element Sample
 SEM/EDS

SEM provides high resolution photos of

conductive and non-conductive materials

When coupled with EDS, it can also provide

elemental composition information

SEM/EDS is semi-quantitative: most quantitative

when the sample is an ideal flat surface of heavy
elements

When the surface is not flat (typical for

contaminants) and comprised of lighter elements
(e.g. C, N, O), it becomes less quantitative

SEM/EDS is ideal for heterogeneous materials

since the EDS spot size can be as small as 1 µm
 SEM/EDS

Fe
Stainless Steel

Fe Cr

Fe
Mn Ni Al

Aluminum
 H o w C a n Yo u H e l p ?

Background information:

• Ingredient list and SDS’s

• H&S handling instructions

• How and when the product was discovered

• Suspected reference samples, such as:

• Labels/packaging container
• Gaskets/tubing
• pump oil
• grease
• mixing blade

• Raw materials as references

 Case Study #1

What is that particle inside of the pre-filled

syringe?

• Upon initial examination we could see a greyish

particle inside of a pre-filled syringe.

• The plastic syringe was cut open and examined

by light microscope. The particle was observed
sitting on the barrel surface.

• We proposed to obtain photos of the particle

and then analyze it by FTIR and SEM/EDS.
 Case Study #1

Light grey particle on syringe wall Magnified

Light microscopy and manual probing showed the

p a r t i c l e t o b e g r e y, o p a q u e a n d e l a s t o m e r i c .
 Case Study #1

Talc

Grey Particle

Butyl
Rubber

FTIR showed the particle to match that of a filled

butyl rubber
 Case Study #1

Si
O
Mg

Analysis by SEM/EDS showed C, O, Mg and Si as

predominant elements
 Case Study #1- Conclusions

The light grey particle is a talc filled butyl rubber

Syringe plunger septum!!!

T h e s e p t u m i s a b l a c k r u b b e r, s o i t i s r u l e d o u t a s
a potential source of the particle

We recommended that the client audit their

manufacturing process with respect to light grey
rubber sources (e.g. gaskets, o-rings, septa)

They could also approach their suppliers as the

particle may have an upstream source
 Case Study #2

What is that floating in the vial?

• During routine QC testing by USP <790>, a

rejected vial was set aside for subsequent
investigation

• Initial examination revealed

a fiber

• We proposed to analyze
the fiber by micro-FTIR
 Case Study #2

Fiber Spectrum

Polystyene Spectrum

Light microscopy- fiber with a flat profile

FTIR analysis identified the fiber as polystyrene.
 Case Study #2- Conclusions

The fiber was identified as polystyrene

The plastic vial was analyzed and found to be

polystyrene

The vial is the likely source

 Case Study #3

What are those particles I can’t even see?

• During QC testing by USP <788> Method 1,

passing results were obtained, however the
counts were higher than expected.

• The product was filtered. Examination of the

filter showed dark particles with sizes in the
range of 25 microns.

• We proposed to analyze the particles by FTIR

and SEM/EDS.
 Case Study #3

We were not able to obtain a useful FTIR

spectrum

The particle is highly absorbing or reflective

 Case Study #3

SEM confirmed the particle sizes

and shapes.

Reference Charcoal

Filtered
Particulate
 Case Study #3

Elemental comparison

Filtered
Particulate

C
O

Reference Charcoal

O
 Case Study #3- Conclusions

B a s e d o n F T I R s p e c t r o s c o p y, S E M a n d E D S
analysis the sub-visible particles were identified
as charcoal

Activated charcoal is used in their processes as a

filtering aid

The manufacturing process did not fully remove

the charcoal from their product

Why??
 Case Study #4

My parenteral is not a clear solution.

Now what?

• Suspensions
• Emulsions
• Implantable Drug products

USP< 1>

“Each final container of all parenteral

preparations should be inspected to the extent
possible for the presence of visible particles”
 Case Study #4

This example will address a parenteral

implantable protein matrix/drug product

An FDA review concluded that the current

inspection of the article surface is not adequate
 Case Study #4

Approach 1
• The protein based product was
digested/dissolved in a caustic solution
• Time, caustic strength and heat were
investigated
• The resulting low viscosity solution was filtered
through a 10 micron stainless steel screen
• Undissolved particulate was quantified as
follows:
• By weight
• Visual examination
(counts, size and description)
• Particles were then identified by FTIR and
SEM/EDS
 Case Study #4

Approach 2
• The protein membrane was hydrated in saline
for 5 minutes to simulate actual OR practice
Saline sample 1
• The hydrated membrane was transferred to
fresh saline and then gently swirled for 15
minutes Saline sample 2
• Both saline solutions were analyzed by USP
<788> Method 1 and 2
• Saline sample 2 represents “loose” particulate
that would be implanted
 Case Study #4

Both approaches were seen as screening

investigations to provide an understanding as to
the number and type of particles involved with
their product

The data was submitted to the FDA

A validated QC method for this product has not

yet been developed and validated
 Concluding Remarks

Analytical characterization of parenteral products

is a common and well understood requirement

The FDA requires definitive physical

characterization of particulate matter in
parenteral products

The objective to minimize and eliminate particles

is based on the ultimate goal of patient safety
 Particulate Matter

Thanks for coming tonight!

Thanks for lending
Jeremiah Masoli to
t h e Ti g e r - C a t s !
Particulate Matter
 Particulate Matter

Questions??