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Workshop Overview
Desmond Hunt, Ph.D.
USP Senior Scientific Liaison
Outline
What are particles?
What is particle size?
Particle size detection and measurement
Commonality
Understanding of particle size
Measurement techniques
What is a particle?
a small object that behaves as a whole unit in
terms of its transport and properties (Wikipedia)
a relatively small or the smallest discrete portion
or amount of something (Merriam-Webster)
a small discrete unit of matter (ASTM)
10
Liquid in liquid
Gas in liquid
11
What is a particle?
Anything that produces a response in the
measurement.
12
Measurement technique
13
Shape
For non-spherical particles, size is not sufficiently
defined by a single number
Leads to measurement variability
Leads to dependence on measurement technique
Pharmaceutically relevant particles exhibit a wide
variety of shapes
14
15
T
h
e
S
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e
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fa
P
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A
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t
t
lin
g
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d
im
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n
t
a
t
io
n
16
Mean
SD
Range
427
489
427
76
33
11
66%
26%
8%
17
Equivalent Sphere
Same min. length
dmin
dw
dmax
Same max.
length
Sphere of same
weight
dv
Same
Volume
dsed
ds
Same sedimentation
rate
dsieve
Same surface
area
18
Volume = p
r h
= 10000
d = 20
20 m
39 m
h = 100
Volume = 4/3
pr
= 10000
r = 39.1
19
Distributions
Particle populations are distributions of both size and
shape.
20
Distributions
Descriptors / Statistics
Basis (y axis)
volume, count, other
Scale (x axis)
linear or logarithmic
Bins
size, resolution
21
22
23
160
140
120
100
80
60
40
20
0
10
100
Diameter (microns)
24
Cumulative
70
60
50
40
30
20
10
0
10
100
Diameter (microns)
25
Log-probability plot
99.9
Cumulative %
99
90
70
50
30
10
1
0.1
10
100
Diameter (microns)
26
Log-probability plot
99
Cumulative %
90
70
50
30
10
1
10
100
Diameter (microns)
27
10
Diameter
28
Relative quantities
1 m
10 m
100 m
count
surface
100
10,000
volume /
mass
1,000
1,000,000
29
Descriptors
Position
Dispersion
Quantiles / Percentiles
30
Descriptors
Means and variances
31
Definitions
Dn or Xn
Qy
GVM
GSD
Width
(D84.13/D15.87)1/2, estimate of
GSD
VMD
Da,b
32
Definitions
d
Qr(x)
33
depends on technique
determines what parameters are used
distribution density
normalize AUC
more correct representation of relative amounts
34
Distribution Density
Linear scale:
q*r,i = Qr,i / xi
35
Interpolation
100
90
80
Q20
70
60
50
40
10
D50
D90
Company Confidential Copyright 2000 Eli Lilly and Company
100
36
x l Du Dl
Dx Dl
u l
Geometric
x l
Dx Dl exp
lnDu / Dl
u l
where
08 Dec 2010 USP Workshop
37
0.18
0.15
0.8
0.12
0.6
0.09
0.4
0.06
0.2
0.03
0
Cumulative Fraction
Volume Fraction
Lognormal Distribution
0
10
100
1000
Diameter
38
Dn x g S
xi x j e
( i j ) ln 2 S
( i j ) ln 2 S
x i x j e x x e(i j ) ln
i
j
39
Sample
Effects / limitations of sampling
Effects of sample preparation
Matrix
40
Sampling
Actual measured sample is (usually) extremely small
fraction of original
Multiple sample divisions chances for bias / segregation
41
42
43
Sample preparation
44
Key thoughts:
45
Measurement Technique
All techniques measure different size related
properties
experimental design
software, algorithms
data handling and reporting
46
Summary of Methods
Microscopy
Image analysis - static and dynamic
Sieving
Electrical sensing zone
Light Diffraction
Light Extinction / Scattering (SPOS)
Dynamic Light Scattering
Aerodynamic methods
Sedimentation
Others
08 Dec 2010 USP Workshop
47
Microscopy
10
20
48
Image analysis
Quantitation of images
Allows more objective use of microscopy
Other uses as well
49
Sieving
Wide overall and dynamic ranges
Slow
Low resolution
Direct mass measurement
50
51
Light diffraction
Relatively wide overall and dynamic ranges
Moderate resolution
Few sampling restrictions
Indirect measurement
Fast
52
53
Sub-micron range
limited resolution
54
Aerodynamic Methods
Impinger
Cascade Impactor
Time of Flight
55
Sedimentation
gravity and centrifugal
photo and X-ray detection
slow
56
Others
Scanning methods
Separation methods
FFF
Ultrasonic attenuation
Laser Doppler velocimetry
57
Summary
Not necessarily only one correct answer
58
Acknowledgements
Some of the slides were provided by Malvern Instruments.
59
Outline
Formulation requirements
Particle properties and powder behavior
Formulation Strategies
Accuracy/Precision dosing
Drug delivery
Manufacturability
Patient acceptability
Stability
Raw Material
Inputs
Manufacturing
Process
Processing
Conditions
Final Product
Outputs
Capsule Filling
Dosator Method
Capsule
Plug
Low density
or fluidized
powder bed
High density
powder bed
Highly consolidated
powder bed
Drug Delivery
Bioavailability can depend on particle size
E.g. Folic acid
Disintegration
Dissolution
Formulation Attributes:
Minimum running characteristics
Compactibility
Fluidity
Lubricity
Release
Particle Properties
PS & PSD
Distribution Type
Particle Shape
10Q 2
2
Aq
x
1 a / x 2
Electrostatic
AL x FA
Capillary
Percolation Segregation
Smaller particles tend to filter down
through larger particles
Bed Vibration
dm SD
(C s Cb )
dt
h
Particle Diameter
S = Surface Area
m = Mass
t = Time
D = Diff. coef.
h = Bndry. layer
C = Conc.
Property Constraints
Flow Rate
Dissolution Rate
Minimum
Flow Rate
Particle Size
Formulation Strategies
Formulation Strategies
The best formulation strategy is?
It depends on circumstances
Need to do risk assessment to decide:
Other excipients, i.e. formulation
Best process to be used
Particle Ranges
Formulation Type
Nano Particles
Size Range
Properties
Micronized API
10 40 m
Typical API
50 150 m
Granules
Beads
Poor fluidity
Poor dissolution
Poor lubricity
np
X np
Assuming p is small thus (1-p) = 1
np (1 p)
%CV - Example
For example:
% CV
20
15
10
5
0
0
250
500
750
1000
Poor compactability
Poor lubricity
Poor fluidity
Poor dissolution
Summary
References
Pharmaceutical dosage forms: tablets, Hoag and Augsburger, 3rd
Ed, vol. 1-3 Marcel Dekker, Inc. (2008)
Presenter
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
87
88
Add-Vantage
Dual Chamber Vials/Syringes
Cartridges, Delivery Pens, and
Autoinjectors
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
89
Parenteral Packaging
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90
Route of Administration
Intravenous
Direct Injection
Infusion
Subcutaneous
Intramuscular
Intradermal
Intraspinal
Intrathecal
Intra-arterial
Others
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
91
92
Regulatory
Meets SISPQ Safety, Identity, Strength, Purity, Quality
Includes sterility, free from pyrogens, toxicologically safe,
essentially particle free
User Requirements
Ease of administration
Pharmaceutical elegance
Comfort
Manufacturing Requirements
Ease of manufacturing
No interactions with processing equipment
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
93
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
94
95
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
96
Requirements
Sterility
Free from microbial contamination for entire shelf life of
product
Tested on a small number of samples compared to the
entire batch size
Methods are Membrane Filtration and Direct Transfer
Tests are limited by:
Sampling and statistical representation
Potential for accidental contamination
Potential for interactions between media, conditions,
and drug that may inhibit microbial growth.
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
97
Requirements
Pyrogen Free
Fever producing agents plus, depending on amount,
can produce a cascade of more serious physiologic
problems including death (sepsis)
Endotoxins: lipopolysacharides from gram negative
microbial contamination
All endotoxins are pyrogens but not all pyrogens
are endotoxins
Limulus Amebocyte Lysate Test (LAL) and rabbit
pyrogen test
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
98
Requirements
Essentially Free from Visible Particulate
Matter
Sources are from throughout the
manufacturing process
Material preparation and handling
Tubing used throughout the process
especially if using a peristaltic pump after
the filtration process
Material used to cover manufacturing
equipment during the autoclave process
Fragments of packaging material used to
contain stoppers & pre-sterilized syringes
Extractables from Bioshield paper used to
cover equipment for autoclaving
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
99
Requirements
Essentially Free from Visible Particulate Matter
Manufacturing
Metal fragments from mixing units
Metal fragments from stopper insertion tubes for prefilled
syringes
Fragments from magnetic stir bars
Metal fragments from abrasion during cleaning processes
Erosion of gaskets
Packaging Materials
Silicone on syringes and stoppers
Glass from breaking ampoules
Coring from puncturing stopper with needle
Removing plastic covering from containers
Dislodging stoppers
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
10
0
10
1
Requirements
Limits for Subvisible Particulate Matter
1st Test by Light Obscuration Method
If the numbers exceed the requirements:
2nd Test by Microscopic Particle Count Method
Test
Product
10 m
25 m
L.O.
SVI
6000
600 / container
LVI
25
3 / mL
SVI
3000
300 / container
LVI
12
2 / mL
Micro
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
User Requirements
Pharmaceutical elegance
Clear, colorless solution
Flows easily through the needle
Little pressure required for injection
Fast rate of administration
Prefer not to stand and slowly administer to patient by
hand
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
10
2
User Requirements
Easy to prepare and stable for user requirements
Prefilled needles ready to inject
Prefilled IV bags
Over pressure in the vial so that it is easier to remove
Syringes and IV infusions may not always be prepared
in laminar flow hoods. This can be a source of
microbial and particle contamination.
Dosages my be prepared 24 hours or more in
advance.
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
10
3
Manufacturing Requirements
Easy to prepare
Requires a limited number of weighing, addition, and pH
adjustment steps
Requires few to no calculations
Requires no in-process release testing
Does not interact with manufacturing equipment
No sticky residues affecting flow through filling needles
M-cresol is absorbed by silicone tubing and will lead to
losses during hold times
Easy to filter
Easy to clean
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
10
4
10
5
Summary
Simple injectable solutions have many rigorous
requirements to ensure patient safety.
2010 Baxter International Inc. Baxter is a registered trademark of Baxter International Inc.
What is Biologics?
Biologics = Biopharmaceuticals (in Europe) ~ Biotherapeutics
Biologics comprise a heterogenous group of pharmaceutical products which are derived from living
organisms, e.g.:
Sugars
Proteins
Nucleic acids
Complex combinations of these substances
May be living entities such as cells and tissues
Biologics
Small Molecules
Chemical entity
Biotherapeutics
MW
< 1 kDa
Half-life
Route of administration
Oral usually
IV or sc usually
Immunogenicity
Toxicology data
(Complexity)
studies
Drug interactions
Very uncommon
Selectivity
Volume of distribution
Active metabolites
None
110
Compound/
function
Coumadin:
BMS;
Biologic*
MW
308.33
MW
(Da)
Compound/
function
204.22
Tryptophan:
Amino acid
Structure
HC
C
C
HC
Anticoagulant
NH
C
HC
CH
NH2
OH
CH
C
CH2
C
O
Amoxil: GSK;
419.45
5808
Bacterial
infection
Singulair:
Merck;
Insulin
(Humulin:Lilly)
Diabetes
608.18
22124
GH deficiency
Lipitor:Pfizer;
Lipid lowering
agent
1209.42
~150000
Representation of
insulin hexamer
structure from
www.biotopics.co.uk/
as/insulinproteinstruc
ture.html
hGH
(Pfizer:
Genotropin)
Asthma
Included only
as a molecular
size reference
(CFK drawing)
mAb
(Humira:
Abbott)
RA
Representation
of human growth
hormone
structure from
Expasy
Representation
of a general
monoclonal
antibody
structure (Pfizer
drawing)
DIA
What about complex synthetic molecules, with or w/o modifications, where there
is heterogeneity?
Oligonuceotides
RNAi
Product-related substances
Molecular variants of the desired product formed during manufacture and/or storage
which are active and have no deleterious effect on the safety and efficacy of the drug
product. These variants possess properties comparable to the desired product and
are not considered impurities.
Example in many mAbs 20-40% deamidated species; various glycosylated forms
Glycoforms
113
114
Ingredient
Purpose
Examples
Surfactant
Reduce aggregate
and particle
formation
Cryo/lyo-protectants
Protect damage
during freezing or
drying
Sucrose, trehalose
Buffers
Maintain pH
Histidine, succinate,
citrate, phosphate
Others as needed
Salts, Antioxidant,
chelators,
preservatives
NaCl, Methionine,
EDTA, phenol/cresol
Excipient addition
Small molecule
API
Mixing/blending
Biologic
Sterile filtration
Compression
Fill/Lyophilize /Finish
Fill/Finish
Oral Tablet
Sterile
Liquid
DIA
LARC/
EIRGM
2008
Sterile Lyophile
Bulk thawing
Prepare diluent
Mix
Add diluent
Mix
IPC: protein conc,
bioburden & pH
Bioburden
reduction
IPC: prefiltrarion
bioburden, conc
Sterile
filter
(0.2m)
Holding
vessel
IPC: Bulk Sterility
Fill &
finish
DIA
LARC/
EIRGM
2008
Biotherapeutics Method
Color
Clarity or Degree of
Opalescence
Visual appearance
Visual Appearance
Visual appearance
Visual Appearance
Visual Inspection
pH
Osmolality
Volume in container
Label Claim
Identity
Purity
Visible Particles
pH
Osmometry
Volume
Absorbance at 280 nm or HPLC
Peptide Mapping(Primary ID method)
CGE (Reducing) / SDS-PAGE
(Reducing) - fragments
CGE (non-Reducing) / SDS-PAGE
(non-Reducing) - aggregate
SEC - aggregate
Binding ELISA
Cell Based Bioassay
Purity
Purity
Potency/Activity
pH
Osmometry
Volume
Safety
Endotoxin
Endotoxin
Safety
Safety
Sterility
Sub-visible Particulate Matter
Sterility
Particulate Matter
Acidic species
(deamidation)
Oxidation
Coloration
Spectrophotometer or using a 96 well plate method at fixed wave-lengths
Comparing to standards
- Munsell color standard
- Gardner solution color standard
- Tristimulus
- Ph. Eur. 2.2.2
Can be an indication of potential interactions
Stir
shake
USP
Stir
shake
IV
+PS80
Visible Particles
Intrinsic
Reversible or irreversible aggregation
Precipitation (solubility, degradants, interactions)
Extrinsic
Environmental particles
Shedding from plastic, stoppers or flakes from glass
Pharmacopoeial Requirements: Extrinsic Particles
USP <1> Injection: essentially free from visible particulates or essentially free from
particles of foreign matter that can be observed on visual inspection
Ph Eur. Injections: Solutions for injection should be clear and practically free from
particles
JP: Injections should meet the requirements of the Foreign Insoluble Matter Test
<6.06> and Insoluble Particulate Matter Test <6.07>. clear and free from readily
detectable foreign insoluble matter
Focus has been on extraneous particles. USP and EP define particulate matter as
extraneous mobile undissolved particles, other than gas bubbles, unintentionally
present in the solutions.
Characterization - Visibility
Clear & essentially particulate-free
Possible presence of non-visible aggregates
Presence of particulates
Size distribution (light obscuration)
Morphological evaluation
Composition analysis after separation
Hazy/turbid
UV
Turbidimeter
~0
<2
<20
<200 <4k
<10k
127 Wei
Wang
Ophthalmic Particles
The USP and JP describe topical ophthalmic product requirements while the Ph.
Eur. does not.
USP 32 <789>: Particulate matter consists of mobile, randomly sourced, extraneous
substances, other than gas bubbles, that cannot be quantitated by chemical analysis
because of the small amount of material they represent and because of their
heterogeneous composition. Ophthalmic solutions should be essentially free from
particles that can be observed on visual inspection. Light obscuration or
microscopic particle count test as per USP <788> are required with more stringent
limits applied.
Drops (per mL basis vs per contianer)
Ointments
Injections
JP XV General Test 18 <6.08>: Ophthalmic solutions prepared as aqueous solution
and aqueous vehicles attached to Ophthalmic Solutions to be prepared before use
should be clear and free from foreign insoluble matter when inspected with the
unaided eye. A limit for Insoluble Particulate Matter is described in JP <6.08>.
Sub-visible particles
Major parts are harmonized across USP, EP and JP though some key
differences remain
1. USP defines large volume injection as those containing >100mL of DP
while Ph. Eur. and JP consider 100 mL product as large volume
injection and requiring a more stringent sub-visible PM limit.
2. Currently, USP exempts injection products intended for subcutaneous
(sc) and intramuscular (im) injection from the requirements of <788>. No
such exemptions are allowed in Ph. Eur and JP, but Ph. Eur. recognizes
that a higher limit may be appropriate for these products.
3. Radiopharmaceutical preparations are exempt from these requirements
and so are the preparations for which the label states that the product is
to be used with a final filter provided that the filter delivers a solution
that complies with the Ph. Eur. sub-visible particles test (2.9.19)
Pharmacopeial Forum article suggesting that the current PM limit
should be more restrictive (not based on extensive biologics product
analysis)
130
Impact of Container/Closure
Intrinsic Particles
At least 12 products have been approved in US and EU where visible protein
particles are generated over time but their amount in relation to the total drug
(protein) is negligible.
Arzerra (ofatumumab) : Colorless solution and may contain a small amount of
visible translucent-to-white, amorphous, ofatumumab particles. However, the
solution should not be used if discolored or cloudy, or if foreign particulate matter is
present.
Vectibix (panitumumab): solution may contain a small amount of visible translucentto-white, amorphous, proteinaceous, panitumumab particulates
Erbitux (cetuximab): solution may contain a small amount of easily visible, white,
amorphous, cetuximab particulates
Stelara (ustekinumab):colorless to light yellow product that may contain a few small
translucent or white particles.
We can expect more!
Time in
Serum at
37C
SEC Results
Weeks
% HMWS
% Main
% LMWS
2.0
97.4
0.6
4.4
92.9
2.7
7.8
86.0
6.2
What do we know?
This is just beginning to be studied. There is lot to learn
Hard to study in-vivo
Need complex analytical (separation, detection) techniques
Can monitor gross changes only
How do these changes link to safety and efficacy?
Acknowledgements
Satish Singh
Carol Kirchhoff
CHARACTERIZATION
DRUG PRODUCT
Appearance
Strength (UV)
IEF
Peptide Map
SDS-PAGE, reducing
SDS-PAGE, nonreducing
HPLC- SEC
Endotoxin
Particulate Matter
Sterility
Binding ELISA
Functional Bioassay
Additional Impurity Assays as needed:
Oxidation
Deamidation (iCE)
Fragment
Identification of Particles
Important to understand the cause
E.g. generation of particles during freeze-thaw
Photo-degradation of the product and generation of particles
Particles
Excipient / drug interactions
When the B2-coated stopper was first used, the xxx drug product formulation
contained polysorbate 80. After the formulation change that removed this surfactant,
fiber-like particles were seen in some final container lots. An investigation into the
phenomenon revealed that a silicon oil coating applied to the stoppers at Mfg site
caused the particles to form in the absence of polysorbate 80. Two development
runs were filled on to establish whether unsiliconized stoppers could be used for
manufacture of drug product that did not contain polysorbate 80. No particles were
observed for these two lots, and the product passed the critical release
specifications. Based on this evidence, the use of unsiliconized stoppers for product
xxx final container production was implemented for all subsequent lots.
146
Outline of Presentation
New technologies for measuring foreign particulate
matter
Advantages and challenges of imaging technologies
Advanced spectroscopic techniques for looking at
isolated particles
Case study
3/25/2011
File name/location
148
Copyright 2010 Eli Lilly and Company
Disclaimer
This information is not intended to suggest
regulatory policy or QC procedures for the release
of clinical trial or market product
3/25/2011
File name/location
149
Copyright 2010 Eli Lilly and Company
Technology used
3/25/2011
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150
Copyright 2010 Eli Lilly and Company
Current Techniques
Advantages
Disadvantages
3/25/2011
File name/location
151
Copyright 2010 Eli Lilly and Company
3/25/2011
File name/location
152
Copyright 2010 Eli Lilly and Company
Detection
Zone
3/25/2011
File name/location
153
Copyright 2010 Eli Lilly and Company
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File name/location
154
Copyright 2010 Eli Lilly and Company
3/25/2011
File name/location
155
Copyright 2010 Eli Lilly and Company
Protein Aggregates
156
Copyright 2010 Eli Lilly and Company
Foreign Particle
3/25/2011
File name/location
157
Copyright 2010 Eli Lilly and Company
Are images
available
Particle
Density
Many particles or
one singular
occurrence
Multiple
Lots
3/25/2011
File name/location
Persistent?
Formulation
Instability?
Glass
incompatibility
Materials
shedding
particles (MOC)
Filters?
Particle
Shape
Sharp
corners: no
solubility
Fluffy
Corrosion
product or
ppt.
Particle
Size
Expect:
Small more
numerous
Large one or
two
Structural
Rigidity
Structural Material
(glass, metal,
polymer, API)?
Residue?
Color
158
Copyright 2010 Eli Lilly and Company
3/25/2011
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159
Copyright 2010 Eli Lilly and Company
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160
Copyright 2010 Eli Lilly and Company
Problem Statement
Development formulation tank was swabbed.
Particulates were found
3/25/2011
File name/location
161
Copyright 2010 Eli Lilly and Company
FTIR Analysis
Preliminary FTIR
analysis
of the particles
showed that
the unknown
material
was PTFE
Swabs were
submitted
for additional
analysis
3/25/2011
File name/location
162
Copyright 2010 Eli Lilly and Company
3/25/2011
File name/location
Element
Wt %
At %
CK
OK
FK
MgK
AlK
SiK
PK
MoL
SK
ClK
KK
TiK
CrK
FeK
NiK
44.32
14.24
06.85
00.53
01.57
05.47
00.67
01.05
00.32
01.94
00.16
01.83
04.96
12.60
03.48
64.34
15.52
06.29
00.38
01.02
03.39
00.38
00.19
00.17
00.96
00.07
00.67
01.66
03.93
01.03
163
Copyright 2010 Eli Lilly and Company
3.1569e+004 max
Company Name
9.34 min
L01336p49B001.spe
x 10
-F1s
Atomic %
C1s
58.0
F1s
25.2
O1s
13.9
Si2p
2.7
Cl2p
0.2
-C1s
Atomic %
C1s 71.5
O1s 28.5
L01336p49I001.spe
x 10
-O1s
2.5
-Cl2s
1200
1000
800
600
Binding Energy (eV)
400
200
0
1400
1200
1000
800
600
Binding Energy (eV)
400
-Cl2p
-Si2s
-Si2p
-F2s
0.5
0
1400
-C1s
c/s
-O KLL
-C KLL
c/s
-O1s
-O KLL
-O KLL
-F KLL2
-F KLL1
-F KLL
1.5
9.34 min
3.5
Company Name
5.1673e+004 max
200
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Copyright 2010 Eli Lilly and Company
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Copyright 2010 Eli Lilly and Company
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Copyright 2010 Eli Lilly and Company
Summary
New technologies will enable more detailed particle
analysis
Numbers of particles in size bins is insufficient for todays drug
development/drug manufacturing arena
Must look at aspects such as:
Frequency of occurrence
Shape
Color
Kinetics where to they appear in the process
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Copyright 2010 Eli Lilly and Company
The Development of
Particle Size Reference Standards
Summary
1. The evolution of particle size standards
2. The importance of producing representative standards
Spatula sampling versus spin riffling
3. The certification of BCR Mirror standards
4. Early laser diffraction results manufacturers and end users
5. A new 19 190 micron opaque microsphere standard
(a) Primary particle size results
(b) Laser results (manufacturers and end users)
Second sample
Contra-rotating
powder disperser
Rotating feeder
Collecting pots
25 Kg
1/100 = 250g
1/100 = 2.5g
Series 1
Series 2
Series 3
Series 4
Series 5
Series Number
1
2
3
4
5
6
Averages
Uncertainty* (+/-)
d (0.1)
37.33
37.63
37.58
37.70
37.54
37.52
37.55
0.26
d (0.5)
63.17
63.36
63.17
63.16
63.08
62.93
63.14
0.28
d (0.9)
92.23
92.25
92.02
92.00
91.87
91.79
92.03
0.36
Tests
57
54
57
55
57
53
3 30 micron standard
Fraunhofer
theory
Mie theory
(2006)
(2006)
10
25
50
75
90
Size (microns)
37.4
46.9
61.0
74.7
87.8
1.0
0.9
0.8
0.7
0.8
Uncertainty (+/-%)
54 tests (Courtesy of LGC)
Mean and uncertainty values for 19 190mm silver coated microsphere standard
Percentile
5
10
25
50
75
90
95
ShapeSizer Size (m)1
Uncertainty (+/-%)
36.02
10.55
40.51
8.49
49.31
5.64
65.80
6.47
72.91
3.20
85.75
4.66
92.65
8.27
34.3
4.52
39.3
4.78
49.1
7.66
64.5
6.98
74.7
6.93
85.6
9.14
92.4
9.59
Malvern Morphologi3
Uncertainty (+/%))
33.9
1.3
38.1
0.84
47.3
1.10
64.6
0.67
73.2
1.69
87.4
1.40
100.4
2.28
1. 5 x 10,000 particles,
2. 5 x 60,000 counts,
3. 5 x 95,000 particles
Notes.
1. The uncertainty levels can be reduced by increasing the particle count
2. Sub-sampling such low corresponding weights may also lead to errors
10
25
50
75
90
95
Sympatec
(Dry)
Beckman (wet)
32.2
37.4
46.9
61.0
74.7
87.8
97.0
34.5
38.8
49.3
63.0
75.7
88.3
97.6
Horiba (wet)
38.1
42.2
50.5
61.3
74.7
88.6
100.1
Horiba (dry)
40.8
44.8
51.9
61.6
73.8
87.2
97.7
Fritsch (wet)
34.1
38.8
48.1
61.1
76.3
91.4
100.6
Malvern (wet)
33.3
38.2
48.6
61.7
75.6
88.2
95.7
37.9
62.2
89.2
5 repeats - Sympatec
(b) Reproducibility
15 labs - PACQS
Comparison of Retsch CAMSIZER and Haver CPA cumulative data with the certificated values
from a 500 2000mm Image Analysis Standard (Multistandard)
Percentile
Certificate data (microns)
Haver CPA (microns)
Retsch CAMSIZER (microns)
5
715
730
721
10
780
790
781
25
897
905
897
50
1101
1125
1123
75
1334
1335
1346
90
1652
1660
1670
95
1816
1827
1846
Reducing count from 150,000 to 15,000 does not significantly affect the results
900nm
380nm
190nm
Inject sample
190nm
380nm
900nm
Conclusions
1. Particle size standards have to evolve to meet the demands
of the latest technology
2. Monosized latex standards for calibrating single channels in a
Coulter Counter are not so suitable for laser sizers or sieves
.
Outline
Overview of reference materials & their properties
Candidate reference materials
Instrumental effects & relationship to reference material
choice
Pathways to accurate measurements
Planned activities
Acknowledgments
Josh Wayment
Germarie Sanchez-Pomales
Michael Carrier
Dick Cavicchi
Rebecca Zangmeister
Michael Tarlov
Vision
Goal 1. Establish repeatability for each instrument type for lowcontrast protein aggregates or surrogates
Diameter
Number
1690
1961
Particle Type
Polystyrene sphere
Polystyrene sphere
Size (m)
1
30
Polystyrene
Spheres
Precip. CaF2 in
CaCl2 solution
Artificial
Particles
Aerogel
fragments
Length / Width
Ratio
Abraded
ETFE
Roughness
Particle concentration
1.10
Apparent error in
concentration, c
1.05
1.00
0.95
0.90
0.90
Error in
size, l
0.95
1.00
1.05
1.10
1.15
Particle size
Characteristic shape
Irregular crystals
1 to 40
Aerogel
Irregular crystals
1 to 20
Irregular, aggregated
polymers
Aggregate of
nanoparticles
Self-assembled polyions
Sintered fumed silica
Natural fibers
Engineered particles
Any 2D shape
<1 to >500
Abraded polymer
Irregular forms
<1 to 100
T or pH induced protein
aggregates
Irregular aggregates;
irregular filaments
<1 to >100
as above
as above
Engineered Fibers
Borrow from semiconductor manufacturing technology:
Advantages
Disadvantages
Any 2D shape
Variable optical density
Variable particle flexibility
Particles are well defined
Possibly expensive
No irregular 3D shapes?
Requires index matching liquid?
ETFE in water + 24 %
sucrose (n = 0.03)
Agitated IgG
Protein Particulates
1. Induce particulate formulation in protein solution by varying
temperature, pH, agitation, etc.
2. Stablize particulates by protein fixative, surfactants, buffer
choice
Are they stable?
Does the particle geometry mimic natural particulates?
Which morphology do we mimic?
(variable scale)
NIST Approach
1. Form aggregates by overnight agitation
Commonalities of instruments:
digital capture and analysis of particulates
flat flow cells
typical range of 1 to 100 m
Proprietary features:
particle identification algorithm
optical light source, camera, objectives, apertures
n = 0.1
0.05
0.02
0.01
Extinction efficiency
0
1
10
Equivalent diameter, m
100
Wavelength = 450 nm
Extinction efficiency
633 nm
800 nm
10
Equivalent diameter, m
100
Extinction efficiency
0.1000
0.0100
0.0010
0.1
1
Equiv. spherical diameter, m
10
Possible Methods:
300
Distance, m
200
100
0
0
20
40
Time, s
60
V/I = R
Perturbing the electrical
resistance R of the channel
... perturbs measured
voltage to current ratio, V/I
Conclusions
Current Standard
Particle
Used
<788>
<789>
Collaborative Testing
12
collaborating Laboratories
Each
Samples
The
Only
the data from the 2nd and 3rd portions were used.
A statistical
12
Counts
On
4000
3500
3000
2500
10 um
15 um
2000
1500
1000
Collaborator 6
500
0
0
10
20
30
40
50
60
Code
70
80
90
100
110
120
3.5
Collaborator 6
10:15 ratio
2.5
1.5
0.5
0
0
10
20
30
40
50
60
Code
70
80
90
100
110
120
10
20
30
40
50
60
70
80
90
100
110
120
1000
500
0
10 um
15 um
-500
-1000
Collaborator 6
-1500
10
20
30
40
50
60
70
80
90
100
110
120
60
40
20
Collaborator 6
-20
-40
-60
10 um
15 um
Lower limit
Upper limit
USP
Track 2: Aerosols
Briggs/Parker/
Marshall/Wiley
Introduction
Essentially Free
NF VII (1942)
Aqueous solutions are to be clear; i.e., when
observed over a bright light, they shall be
substantially free from precipitate, cloudiness, or
turbidity, specks or flecks, fibers or cotton hairs,
or any un-dissolved material. Substantially free
shall be construed to mean a preparation which
is free from foreign bodies that would be readily
discernable by the unaided eye when viewed
through a light reflected from a 100 watt Mazda
lamp using as a median a ground glass and a
background of black and white
USP XX (1980)
Every care should be exercised in the
preparation of injections to prevent
contamination. Good pharmaceutical practice
also requires that each injection, in its final
container, be subjected individually to a
physical inspection, whenever the nature of
the container permits, and that every
container whose contents show evidence of
contamination with visible foreign material be
rejected. [Same as XIX Supplement 1]
PF Stimuli Proposal
Based on Fed. Std. 142a, updated AQL from date
captured by PDA Survey of Visual Inspection
Practices 2008
Attempts to harmonize USP, JP and Ph. Eur.
Stakeholders comments captured at May 13,
2010, meeting of USP Visual Inspection of
Parenterals Advisory Panel
Comments are being addressed and will likely
result in several changes, including the proposed
AQL and sampling levels
Parenteral Particles
(Physical Characteristics)
Soft Particles
refers to droplets, as, e.g., emulsions, such as
injectable oil-in-water dispersions
flexible, malleable, deformable
LOW embolic risk
Hard Particles
refers to solids, particulates, precipitates in
injectable liquids
inflexible, rigid, obstructing
HIGH embolic risk
Metabolic Risk
Blood flow from the left side of the heart
into the aorta main arteries to vital organs
Immunological Risk
Intravenous route for allergen increases the
risk for systemic anaphylactoid response,
irrespective of particle hardness or softness
Pharmacological Risk
Alteration of homogeneity of dosage form can
result in therapeutic failures from either a
subtherapeutic (under-delivery) or toxic
(over-delivery) response.
MEDICAL RISKS:
Particulates and Embolic
Phenomena
1996;20:81-87
(CaHPO4)
Report Summary
Paroxysmal respiratory failure and death
occurred in two young adult females
autopsy revealed an amorphous material
containing containing calcium [and
phosphorus] obstructing the pulmonary
microvasculature of each patient both
received an identical [intravenous]
admixture.
DFD-SSLLC-2010
MEDICAL RISKS:
Lipids and Embolic Phenomena
Report Summary
The pathophysiological effects of infusing
large amounts of stable lipid emulsions
have been shown to cause RES
dysfunction in animals and humans. This
study investigated the effects on vital RES
organs, i.e., liver and lungs, from the
infusion of a stable (PFAT5 < 0.05%) vs.
highly unstable (PFAT5 ~ 50x higher) fat
infusion over 24 hours.
Characterization
FAT DEPOSITS:
Normal (absent); Abnormal (limited, moderate or diffuse)
TISSUE STRUCTURE:
Normal (intact); Abnormal ( cellularity, infiltration, and/or necrosis)
MEDICAL RISKS:
Lipids and Injury
to Vital Organs
Report Summary
The pathophysiological effects of infusing
large amounts of stable lipid emulsions
have been shown to cause RES
dysfunction in animals and humans. This
study investigated the effects on vital RES
organs from the infusion of a stable
(PFAT5 < 0.05%) vs. modestly unstable
(PFAT5 ~ 10x higher) fat infusion over 24
hours.
MEDICAL RISKS:
Lipids and Impaired Metabolic
Clearance
J Pediatrics 2008;152:232-36
MEDICAL RISKS:
Proteins and Particulates and
Immunogenic Phenomena
Safety-Related
Regulatory Actions:
41/174 = 23.6%
James R. Coleman
John A. Robson
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50 particles/mL
Particles 25 m
5 particles/mL
Particles 50 m
2 particles/mL
Foreign Particulate Matter in OINDP: General Considerations
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Composition:
Health: Safety assessment
Specific compositions
Particle number
Source: identification for control
Foreign Particulate Matter in OINDP: General Considerations
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Number of
particles
analyzed = N
N ~ Std. Dev.
% RSD
10
3.16
31.62%
100
10.00
10 %
1,000
31.62
3.16
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Microscopy
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10.00
8.00
6.00
Titanium dioxide density
4.20
4.00
2.00
0.00
0.00
20.00
30.00
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as
foreign particulates
Polybutylene Terephthalate
Aluminum
Polycarbonate
Bacterial fragments
Polychlorotrifluoroethylene
Bromobutyl rubber
Polyester
Clay
Polyethylene
Copper
Polyimide
Elastomer
Polyoxymethylene
Polypropylene
Glass
Stainless steel
Iron
Talc
Kaolin
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The End
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The Beginning
of an interesting journey
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For OINDP:
Prior to use
Open the device
(carefully)
To distinguish functional
source
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Scope
Focus on foreign particulate in aerosolized drug
products
Safety evaluation
Considerations for establishing safety limits
Sources and types
Clinical safety
Slide 345
Slide 346
Total
0.8
Head
0.6
0.4
Tracheobronchial tree
0.2
Pulmonary
0
0
10
15
20
Slide 350
Slide 352
Slide 354
Slide 355
Slide 356
Slide 357
Diameter, m
1
3.6 x 107
4.5 x 106
1.1 x 107
1.3 x 106
4.5 x 106
5.6 x 105
1.3 x 106
1.7 x 105
5.6 x 105
7.0 x 104
10
2.9 x 105
3.6 x 104
Slide 360
Slide 361
Slide 362
Slide 363
polybutylene terephthalate
polyethylene
polyester
polyoxymethylene
polyimide
polystyrene
polypropylene
stainless steel
skin cells
talc
transparent synthetic fibers
Slide 365
Slide 367
Slide 369
Slide 372
Slide 373
Conclusions
Safety limits can be developed once foreign
particulates have been characterized in terms of
number and identity
Safety limits for foreign particulates 10 m are
most appropriate since these can penetrate into
the lungs
Establishing safety limits based on 1-5% of the
EPA NAAQS PM10 (30-150 g/day) should
provide adequate protection for most types of
foreign particulates, but not necessarily all.
Slide 375
Acknowledgements
IPAC-RS Foreign Particulate Working Group
Courtney Crim, MD, GSK for clinical safety
assessments
Ron Wolff, PhD, Novartis for development of the
NAAQS safety assessment
Slide 376
References
Slide 377