Best Practices for OINDP Pharmaceutical
Development Programs
Leachables and Extractables
V. The Controlled Extraction Study
PQRI Leachables & Extractables Working Group
PQRI Training Course
September 20-21, 2006
Washington, DC
Definition
►A
Controlled Extraction Study is a laboratory
investigation into the qualitative and
quantitative nature of extractable profiles
from critical components of an OINDP
container/closure system
PQRI Safety Thresholds and Best Practices
for Extractables and Leachables in OINDP
November 2005
Top Ten Reasons Why Controlled
Extraction Studies are Needed
►
►
►
►
►
To make an informed selection
of materials.
To meet regulatory
expectations.
To control leachables.
To control materials from lot to
lot.
To correlate extractables data
to leachables.
►
►
►
►
►
To evaluate the safety of the
materials.
To predict worst case of endof shelf life leachables.
To qualify packaging
materials.
To obtain a comprehensive
extractables profile.
Because USP testing does
not provide applicable data.
Course Objectives
► Purpose
of a Controlled Extraction Study
► PQRI Best Practice Recommendations
 Controlled Extraction Study Example Data
►Qualitative
and Quantitative Profiles
►Method Optimization
► Conclusion
The Purpose of a Controlled Extraction Study is
to systematically and rationally identify and
quantify potential leachables, to the extent
practical, and within certain defined analytical
threshold parameters.
Utility of Extractable
Information
► Obtain
Data for Risk Assessment
 Provide Information to Toxicologists for
Preliminary Risk Assessment
 Apply Threshold Principles
► Provide
Basis for Leachable Methods
► Correlate Extractable Data to Leachables
Data
► Develop Routine Extractable Tests
► Establish Control Criteria
Study Strategy
► When
to Begin
 Early in Development Phase
► Establish
► Where
Team and Obtain Extractable Information
to Begin
 Select Critical Components
► Knowledge
of Materials
 Extraction Solvents/Techniques
 Analytical Methods
► Where




to End
Application of the AET
Identification Categories
Data Evaluation and Reporting
Control of Leachables
Critical Components
► MDI,
DPI, Nasal Inhalation Solutions and
Sprays





Patient Contact
Product Contact
Device Performance
Secondary Packaging
Ancillary Components
Typical Materials
► OINDP









Components
Valves (Gaskets/O-rings)
Mouthpiece
Canister
Secondary Packaging
Pump Components
Actuator
Containers
Blisters
Labels/Adhesives/Inks
►
Extractables (0.01-1000ug)










Solvents
Monomers/Dimers/Trimers
Curatives
Photo Initiators
Plasticizers
Lubricants
Processing Aids
Antioxidants
Cleaning Residues
Reaction/Degradation and
Breakdown Products
Knowledge of
Materials/Processes
► Materials
of Composition
 Base Material
 Additives and Processing Aids
► Polymerization
Process
► Fabrication process
► Cleaning and Pretreatment
► Component Storage and Shipping
Extractable Profiles
► Qualitative
 Comprehensive
► Quantitative
 Worst Case Leachables
► Component
Control
 Acceptance Criteria
Challenges/Choices
• What Components?
• How Many Components ?
• What Volume of Solvent?
• What Reference Material Should be
Selected?
• What Solvents?
• What Extraction Techniques?
• What Analysis Conditions?
Considerations
► Extraction
should be vigorous, but not so
aggressive as to alter the qualitative and/or
quantitative nature of the extractable profile
► Must be technically justified and optimized
to produce extractables profiles at least
equivalent to leachable profiles obtained
under worst case conditions
Jenke, DR. PDA J Pharm Sci Technology,
2003
PQRI Best Practices
Recommendations
Ten Principle Objectives
#1 Controlled Extraction Studies should
employ vigorous extraction with
multiple solvents of varying polarity.
► Range
of Polarities
► Range of Boiling Points
► One of Similar Extracting Properties to Drug
Product Vehicle
► Relatively Non-reactive
► High Purity
► Easily and Safely Handled
► Readily Available
Solubility
DAD1 A, Sig=200,4 Ref=550,100
mA (I:\HPCHEM\1\DATA\022569\022569\JAN31008.D)
U
DAD1 A, Sig=200,4 Ref=550,100 (I:\HPCHEM\1\DATA\022569\022569\JAN31012.D)
mAU
300
17
5
250
15
0
2-propanol
(Reflux)
Hexane (Reflux)
12
5
200
10
0
150
75
100
50
50
25
0
0
0
2
4
6
8
10
12
14
16
18 min
0
2
4
6
8
10
12
14
16
18 mi
Thermolysis
A b u n d a n ce
A bundance
A bundance
T IC : 0 2 2 1 0 3 0 2 .D
T IC : 0 2 2 7 0 3 0 3 .D
T IC : 0 2 2 5 0 3 0 3 .D
1 .1 e + 0 7
1 .2 e + 0 7
1 .2 e + 0 7
1 .1 5 e + 0 7
1 .1 5 e + 0 7
1 .1 e + 0 7
1 .1 e + 0 7
1e+07
1 .0 5 e + 0 7
1 .0 5 e + 0 7
9500000
1 .0 5 e + 0 7
1 e + 0 7
1e+07
9000000
9 5 0 0 0 0 0
9500000
8500000
9 0 0 0 0 0 0
9000000
8000000
8 5 0 0 0 0 0
8500000
8 0 0 0 0 0 0
8000000
7 5 0 0 0 0 0
7500000
7 0 0 0 0 0 0
7000000
6 5 0 0 0 0 0
6500000
6 0 0 0 0 0 0
6000000
5 5 0 0 0 0 0
5500000
5000000
5 0 0 0 0 0 0
5000000
4500000
4 5 0 0 0 0 0
4500000
4000000
4 0 0 0 0 0 0
4000000
3500000
3 5 0 0 0 0 0
3500000
3000000
3 0 0 0 0 0 0
3000000
2 5 0 0 0 0 0
2500000
2 0 0 0 0 0 0
2000000
1 5 0 0 0 0 0
1500000
1 0 0 0 0 0 0
1000000
5 0 0 0 0 0
500000
7500000
7000000
6500000
6000000
5500000
2500000
2000000
1500000
1000000
500000
0
0
5 .0 0
1 0 .0 0
1 5 .0 0
2 0 .0 0
2 5 .0 0
T im e - - >
3 0 .0 0
3 5 .0 0
5 .0 0
1 0 .0 0
1 5 .0 0
2 0 .0 0
Methylene Chloride
2 5 .0 0
3 0 .0 0
3 5 .0 0
5 .0 0
1 0 .0 0
1 5 .0 0
T im e -->
T im e - - >
2-propanol
Hexane
2 0 .0 0
2 5 .0 0
3 0 .0 0
3 5 .0 0
Extractable Yield
Abundance
Abundance
TIC: 02280334.D
1300000
2000000
TIC: 02280342.D
Abundance
TIC: 02280338.D
1800000
1200000
1800000
1100000
1600000
1600000
1000000
1400000
1400000
900000
1200000
800000
1200000
1000000
700000
1000000
600000
800000
800000
500000
600000
400000
600000
400000
300000
400000
200000
200000
200000
Time-->
100000
0
5.00
10.00
15.00
20.00
25.00
30.00
Methylene Chloride
35.00
Time-->
0
0
5.00
10.00
15.00
20.00
2-propanol
25.00
30.00 Tim35.00
e-->
5.00
10.00
15.00
Hexane
20.00
25.00
30.00
35.00
#2 Controlled Extraction Studies should
incorporate multiple extraction
techniques.
Sulfur Cured Elastomer
A b u n d a n c e
A bundance
Abundance
T IC : 0 3 1 4 0 3 0 3 .D
T IC : 0 2 0 3 0 3 0 4 .D
4500000
TIC: 02270303.D
1 .3 e + 0 7
1.2e+07
1 .2 5 e + 0 7
1.15e+07
1 .2 e + 0 7
1.1e+07
4000000
1 .1 5 e + 0 7
1.05e+07
1 .1 e + 0 7
1e+07
1 .0 5 e + 0 7
3500000
9500000
1 e + 0 7
9000000
9 5 0 0 0 0 0
8500000
9 0 0 0 0 0 0
3000000
2500000
2000000
8 5 0 0 0 0 0
8000000
8 0 0 0 0 0 0
7500000
7 5 0 0 0 0 0
7000000
7 0 0 0 0 0 0
6500000
6 5 0 0 0 0 0
6000000
6 0 0 0 0 0 0
5500000
5 5 0 0 0 0 0
5000000
5 0 0 0 0 0 0
1500000
4500000
4 5 0 0 0 0 0
4000000
4 0 0 0 0 0 0
3500000
3 5 0 0 0 0 0
3000000
1000000
3 0 0 0 0 0 0
2500000
2 5 0 0 0 0 0
2000000
2 0 0 0 0 0 0
500000
1500000
1 5 0 0 0 0 0
1000000
1 0 0 0 0 0 0
500000
5 0 0 0 0 0
0
5 .0 0
1 0 .0 0
1 5 .0 0
T im e - - >
2 0 .0 0
2 5 .0 0
3 0 .0 0
3 5 . 00 0
0
5 .0 0
1 0 .0 0
1 5 .0 0
2 0 .0 0
T im e - - >
Sonication
Soxhlet
2 5 .0 0
3 0 .0 0
3 5 .0 0
Time-->
5.00
10.00
15.00
Reflux
20.00
25.00
30.00
35.00
Peroxide- Cured Elastomer
Abundance
Abundance
TIC: 02280309.D
TIC: 02280324.D
1000000
4000000
900000
3500000
800000
700000
3000000
600000
2500000
500000
2000000
400000
1500000
300000
1000000
200000
500000
100000
Time-->
0
5.00
10.00
Sonicatrion
15.00
20.00
25.00
30.00
35.00
0
Time-->
5.00
10.00
Reflux
15.00
20.00
25.00
30.00
35.00
Polypropylene
DAD1 A, Sig=200,4 Ref=550,100 (J:\HPCHEM\1\DATA\022569\FEB14011.D)
DAD1 A, Sig=200,4 Ref=550,100 (I:\HPCHEM\1\DATA\022569\022569\JAN31008.D)
mAU
175
mAU
150
175
2-propanol Blank (Sonication)
125
100
150
2-propanol (Reflux)
75
50
125
25
0
0
2
4
6
8
10
DAD1 A, Sig=200,4 Ref=550,100 (J:\HPCHEM\1\DATA\022569\FEB14012.D)
12
14
16
18 min
mAU
175
100
75
022569-01 2-propanol Extract (Sonication)
150
Ultranox 626 peak 1
125
100
75
50
25
50
25
0
0
0
2
4
6
8
10
12
14
16
18 min
0
2
4
6
8
10
12
14
16
18 min
#3 Controlled Extraction Studies should
include careful sample preparation
based on knowledge of analytical
techniques to be used.
► Preparation
of Extracts
 Sampling, Sample:Surface Ratio, Solvents,
Conditions
► Test
Sample Preparations
 Instrumental Techniques
 Concentration/Dilution
Sample Introduction
Abundance
A b u n d a n c e
TIC: 02270303.D
T IC : 0 2 2 7 0 3 0 2 .D
1 .2 e + 0 7
1.2e+07
1 .1 5 e + 0 7
1.15e+07
1 .1 e + 0 7
1.1e+07
1 .0 5 e + 0 7
1.05e+07
1 e + 0 7
1e+07
9 5 0 0 0 0 0
9500000
9 0 0 0 0 0 0
9000000
8 5 0 0 0 0 0
8500000
8 0 0 0 0 0 0
8000000
7 5 0 0 0 0 0
7500000
7 0 0 0 0 0 0
7000000
6 5 0 0 0 0 0
6500000
6000000
6 0 0 0 0 0 0
5500000
5 5 0 0 0 0 0
5000000
5 0 0 0 0 0 0
4500000
4 5 0 0 0 0 0
4000000
4 0 0 0 0 0 0
3500000
3 5 0 0 0 0 0
3000000
3 0 0 0 0 0 0
2500000
2 5 0 0 0 0 0
2000000
2 0 0 0 0 0 0
1500000
1 5 0 0 0 0 0
1000000
1 0 0 0 0 0 0
500000
5 0 0 0 0 0
0
5.00
5 .0 0
1 0 .0 0
1 5 .0 0
2 0 .0 0
2 5 .0 0
3 0 .0 0
3 5 .0 0
10.00
15.00
20.00
25.00
30.00
Time-->
T im e -->
2- Propanol Extract
Reconstituted in Methylene Chloride
35.00
Artifacts
► 2-(chloromethylthio)benzothiazole
Heat
N
SH
S
+
N
S
CH2Cl2
S
Cl
+
HCl
Solvent Compatibility
DAD1 A, Sig=200,4 Ref=550,100 (G:\HPCHEM\1\DATA\022569\JAN31009.D)
mAU
175
150
Methylene Chloride Blank (Reflux)
125
100
75
50
25
0
mAU
175
12
Ultranox 626 peak 1
022569-01 Methylene Chloride (Reflux)
150
125
100
75
50
14
16
18
min
14
16
18
min
Irganonx 1010
0
2
4
6
8
10
DAD1 A, Sig=200,4 Ref=550,100 (G:\HPCHEM\1\DATA\022569\JAN31010.D)
25
0
0
2
4
6
8
10
12
Solvent Compatibility
#4 Controlled Extraction Studies should
employ multiple analytical techniques
►
Gas Chromatography/Mass Spectrometry (GC/MS)
►
Liquid Chromatography/Mass Spectrometry (LC/MS)
►
Liquid Chromatography/Diode Array Detection (LC/DAD)
►
Gas Chromatography/Flame Ionization Detection (GC/FID)
►
Liquid Chromatography/Ultraviolet Detection (LC/UV)
►
Fourier Transform Infrared Spectroscopy (FTIR)
►
Inductively Coupled Plasma/Mass Spectroscopy (ICP/MS)
►
Inductively Coupled Plasma/Optical Emission Spectroscopy
(ICP/OES)
►
Scanning Electron Microscopy. Energy Dispersive X-Ray
(SEM/EDX)
Qualitative System Suitability
Compound
Suggested
Technique
Recommended Target
Concentration (ug/ml)
GC or LC
50
GC or LC/UV
50
GC or LC
50
Irganox 1010
LC
50
Diphenyl Amine
LC
50
Bis-(2-ethylhexyl) phthalate
GC or LC
50
Bis (dodecyl) phthalate
GC or LC
50
GC or LC/MS
100
GC
50
GC or LC/UV
1
2-Mercaptobenzothiozole
Tetramethylthiuramdisulfide
Butylatedhydroxytoluene
Stearic Acid
2-ethylhexanol
Pyrene
Complimentary Techniques
A b u n d a n c e
T IC : J A N 3 1 1 0 8 .D
1 1 0 0 0 0
2 - p r o p a n o l R e f lu x
1 0 0 0 0 0
9 0 0 0 0
3
2
1
8 0 0 0 0
7 0 0 0 0
6 0 0 0 0
5 0 0 0 0
4 0 0 0 0
3 0 0 0 0
2 0 0 0 0
1 0 0 0 0
0
5 .0 0
1 0 .0 0
1 5 .0 0
2 0 .0 0
2 5 .0 0
3 0 .0 0
3 5 .0 0
T im e - - >
GC/MS of polypropylene 2-propanol extract: 1 = 2,6-di-methyl benzaldehyde;
2 = 2,4-di-tert-butylphenol; 3 = glycerol monostearate.
Complimentary Techniques
5.3
8.6
10.6
15.6
16.0
Bis(dimethylbenzylidene)
sorbitol isomer
Unknown
Di-tert-butylphenol
Tetradecanoic acid
Hexadecanoic acid
18.4
19.0
19.4
20.3
21.0
Glycerol monopalmitate / Glycerol
monostearate
Irganox 1010 fragment
Irganox 1010 related
Octadecanoic acid
Irganox 1010
Compound Specific Detection
Reflux PP Disc/CH2Cl2
03270311
280 nm
ANALOG
5.22e5
100
%
0
03270311
Scan AP1175
2.93e6
100
%
0
Time
5.00
10.00
15.00
20.00
25.00
30.00
35.00
40.00
HPLC-UV chromatogram and m/z 1175 extracted ion current profile
#5 Controlled Extraction Studies should
include a defined and systematic
process for identification of
individual extractables
Abundance
TIC: 02050302.D
1.35e+07
1.3e+07
1.25e+07
1.2e+07
35
1.15e+07
1.1e+07
1.05e+07
1e+07
36
9500000
9000000
41
8500000
8000000
33
7500000
45
7000000
6500000
31
6000000
49
5500000
5000000
51
4500000
4000000
30
3500000
53
3000000
2500000
55
2000000
1500000
1000000
500000
0
5.00
Time-->
10.00
15.00
20.00
25.00
30.00
35.00
Identification Categories
► Confirmed:
 Mass spectrometric fragmentation behavior
 Confirmation of molecular weight or confirmation of elemental
composition
 Mass spectrum matches automated library or mass spectrum
and chromatographic retention index match authentic specimen
► Confident:
 Sufficient data to preclude all but the most closely related
structures have been obtained
► Tentative:
 Data have been obtained that are consistent with a class of
molecule only
Confirmed Peak 21.47 min
2,2’-methylene-bis-(-6-tert-butyl)-4-ethylphenol
Confirmation of Molecular Weight; Fragmentation Behavior;
Mass Spectral Library Match; RT Match to Authentic Standard
Abundance
TIC: 02050302.D
1.35e+07
1.3e+07
1.25e+07
1.2e+07
35
1.15e+07
1.1e+07
1.05e+07
1e+07
36
9500000
9000000
41
8500000
8000000
33
7500000
45
7000000
6500000
31
6000000
49
5500000
5000000
51
4500000
4000000
30
3500000
53
3000000
2500000
55
2000000
1500000
1000000
500000
0
5.00
Time-->
10.00
15.00
20.00
25.00
30.00
35
Confirmed Peak 21.47 min
2,2’-methylene-bis-(-6-tert-butyl)-4-ethylphenol
T071703005 1173 (21.561) Cm (1171:1177-(1164:1168+1201:1214))
88
100
Scan CI+
4.35e3
100
74
[M+NH4]+
CI mass spectrum (ammonia)
%
130
386
208
86
70
61
73
84
101
128
98
114
124
135
163
152 161
177
175 178
0
F07013003 1111 (21.557) Cm (1109:1112-1116:1122)
100
191
192
209
193
218
229 237
252 257
272 274 276 293
313
312
314
330
331
387
368 370
352 353 367
377
388 397
409
425
MS2 EI+
4.86e4
191
163
420
175 178
EI mass spectrum
M+. ????
%
135
368
141
57
91
65
0
60
77
69
79
80
119
105
117
103
147 155
133
121
120
192
312
179
180
85
100
169
140
160
180
193 202
200
256
215
220
227 237
240
255
257 267 281 295
260
280
297 311 313 321
300
320
369
339
353 354
340
360
370
380
m/z
400
420
Confident Peak 8.15 min
Benzothiazole
Mass Spectrometric Fragmentation Behavior; Mass Spectrum
Matches Automated Library Search
Abundance
TIC: 02270303.D
1.2e+07
1.15e+07
1.1e+07
1.05e+07
1e+07
9500000
9000000
8500000
8000000
7500000
7000000
6500000
6000000
5500000
5000000
4500000
4000000
3500000
3000000
2500000
2000000
1500000
1000000
500000
0
5.00
Time-->
10.00
15.00
20.00
25.00
30.00
35.00
Confident Peak 8.15 min
Benzothiazole
Abundance
Average of 8.145 to 8.166 min.: 02050302.D (-)
135
9000
8000
7000
6000
5000
4000
108
3000
2000
69
1000
50
0
30
40
50
58
82
63
60
91
74
70
80
90
100
110
120
130
140
m/ z-->
Abundance
#23599: Benzothiazole (CAS) $$ Vangard BT $$ 2-BENZOTHIAZO...
135
9000
8000
7000
6000
5000
4000
108
3000
2000
69
1000
39
0
30
m/ z-->
40
45 50
50
82
58 63
60
91
74
70
80
90
100
110
120
130
140
Tentative Peak 15.05 min
Coumarone-indene resin related
Confirmation of Elemental Composition; Mass Spectrometric
Fragmentation Behavior
Abundance
TIC: 02270303.D
1.2e+07
1.15e+07
1.1e+07
1.05e+07
1e+07
9500000
9000000
8500000
8000000
7500000
7000000
6500000
6000000
5500000
5000000
4500000
4000000
3500000
3000000
2500000
2000000
1500000
1000000
500000
0
5.00
Time-->
10.00
15.00
20.00
25.00
30.00
35.00
Tentative Peak 15.05 min
Coumarone-indene resin related
A b u n d a n c e
A v e ra g e
o f 1 5 .0 3 6
1 1 9
to
1 5 .0 6 8
m in . : 0 2 0 5 0 3 0 2 . D
(-)
+
3 4 0 0 0
3 2 0 0 0
3 0 0 0 0
2 8 0 0 0
2 6 0 0 0
2 4 0 0 0
+
2 2 0 0 0
+
2 0 0 0 0
+
1 8 0 0 0
1 6 0 0 0
9 1
1 4 0 0 0
+
1 2 0 0 0
1 0 0 0 0
8 0 0 0
6 0 0 0
4 0 0 0
7 7
1 0 3
2 0 0 0
5 1
6 5
0
5 0
m / z -->
6 0
2 3 6
1 2 8
7 0
8 0
9 0
1 4 5
1 6 5
1 8 0
1 8 9
2 0 2
2 2 1
1 0 0 1 1 0 1 2 0 1 3 0 1 4 0 1 5 0 1 6 0 1 7 0 1 8 0 1 9 0 2 0 0 2 1 0 2 2 0 2 3 0 2 4 0
#6 Controlled Extraction Study
“definitive” extraction methods should
be optimized.
► Asymptotic
Levels
► Represent at Least Worst Case Leachables
 Qualitative
 Quantitative
► Verification
of Quantitative Results
► Basis for Development and Validation of
Routine Extractable Control Methods
Sulfur Cured Rubber
7g/200ml
Methylene Chloride
10:1 Dilution
Internal Standard
16 Hours Extraction
1
Area ratio, analyte to internal standard
Analysis:
GC/MS
Extraction: Soxhlet
Optimization:
0.9
0.8
0.7
Phenolic
0.6
Docosane
Hexacosane
0.5
Coumarone indene
0.4
0.3
0.2
0.1
0
0
2
4
6
8
10
Time, hours
12
14
16
18
Polypropylene
Extraction:
Reflux
Analysis:
HPLC/UV
Optimization:
Solvent to Sample Ratio
Analyte Solubility/Standardization
Asymptotic Extraction
Chromatography Conditions
Results:
1g:25ml (50/50 THF/IPA)
3 Hr Extraction
Polypropylene
Polypropylene
1
6
0
0
1
4
0
0
1
2
0
0
1
0
0
0
Concetraion,ug/
8
0
0
6
0
0
4
0
0
2
0
0
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3
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6
7
Example Chromatography Conditions
► Qualitative
► Quantitative
LC/MS
HPLC/DAD
Column:
C18, 4.6mmx25cm, 5u
Injection Vol: 10ul (1/mL/min)
Mobile Phase: <A> 75:25 (ACN:H20)
<B> 50:50 (ACN:THF)
Gradient:
30 min. gradient at 100%
A to 100% B hold 12 min
Ionization:
APCI
Scan:
m/z 50-1350 ,5 sec/scan
Column:
C18, 4.6mmx25cm, 5u @
60C
Injection Vol: 10ul (1/mL/min)
Mobile Phase: <A> ACN
<B> H2O
Gradient:
25 min. linear gradient at
30:70 A:B to 100% A
hold 5min
Detector:
Diode Array Detector
200nm, 220nm, bw/4nm;
ref.sig 550nm, bw/100nm
Quantitation
Internal Standard
RRF = (AaCi)/(AiCa)
 Aa= Area of Analyte Peak
 Ci = Concentration of Internal
Standard
 Ai = Area of Internal Standard
Peak
 Ca = Concentration of Analyte
►
External Standard
RF = Cs/As
 Cs = Concentration of External
Standard
 As= Area of External Standard
►
Ca= Aax RF
Concentration in Sample Extract
Total mass = Caug/ml x Vol of extract
Ca= Aax Ci/Aix RRF
►
Total mass = Caug/ml x Vol of extract
►
Extractable in Component
μg/g Extractable =
Total mass (μg)/g Component
Concentration in Sample Extract
Extractable in Component
μg/g Extractable =
Total mass (μg)/g Component
System Suitability Example
Instrument Precision (%RSD)
≤ 10
Resolution (n=6)
≥2
Tailing Factor (n=6)
≤2
Sensitivity (S/N of MQL)
≥ 10
Method Repeatability (%RSD)
≤ 10%
Intermediate Precision (%RSD)
≤ 10%
Recovery
80-120%
Method Validation
► Acceptance






Criteria
System Suitability
Instrument Precision
Chromatographic Resolution
Chromatographic Tailing Factor
Linearity/Range
Precision
► Method
Repeatability
► Standard Sample Stability





Intermediate Precision
Specificity
Accuracy
Limit of Quantitation/Limit of Detection
Robustness/Ruggedness
#7 During the Controlled Extraction
Study process, sponsors should
revisit supplier information
► Investigate
 Chemical entities found in the extractable data
not included in supplier information
 Known chemical entities not detected in the
extractable
Extractable Data
Code
Letter
Qualitative Results
Supplier
Information
Quantitative
Results
C
Tetrakis (methylene(3,5-di-t-butyl-4hydroxyhyrocinnate)) methane
Phenolic Antioxidants
0.08 %
0.059%
B
Bis(2,4-di-t-butylphenyl)pentaerythritol
disphosphite and di-tert butylphenol
Phosphite Antioxidant
0.05%
0.045%
Calcium Stearate
Stearate Mould Release
0.03 – 0.4%
Not
Detected
Glycerol monopalmitate/monostearate
Vegetable oil
0.2 - 0.3 %
Not
Detected
Tetradeconoic, Hexadecanoic and
Octadecanoic Acids
N/A
N/A
Clarifier
0.2 - 0.3 %
0.14
A
3,4-dimethyldibenzylidene sorbitol
Known Accelerator
Abundance
TIC: 02050302.D
1.35e+07
1.3e+07
1.25e+07
Abundance
1.2e+07
35
1.15e+07
TIC: 02030503.D
1.1e+07
900000
1.05e+07
Tetramethylthiuram Monosulfide
1e+07
800000
9500000
36
9000000
41
700000
8500000
8000000
33
600000
7500000
45
7000000
6500000
500000
31
6000000
49
5500000
400000
5000000
51
4500000
300000
4000000
30
3500000
200000
53
3000000
2500000
100000
55
2000000
1500000
0
1000000
5.00
10.00
15.00
20.00
25.00
30.00
35.00
5.00
10.00
15.00
20.00
25.00
30.00
35.00
Time-->500000
0
Time-->
GC-MS of Methylene Chloride Soxhlet Extract
#8 Controlled Extraction Studies should
be guided by an Analytical
Evaluation Threshold (AET) that is
based on an accepted safety
evaluation threshold
► How
low to go to identify and evaluate
individual extractable
AET Extractables
►A
leachable dose less than or equal to the
SCT is a dose so low that there would be
negligible safety concerns from toxic effects.
 Internal or External Standards can be employed
to Measure the SCT level.
 The sensitivity needed for the extractable and
leachable methods can be postulated.
 Comprehensive extractable studies can be
predictive of end of shelf life leachable studies
1st Application of the Analytical
Evaluation Threshold (AET)
► Example
Nasal Spray Component #1:
4 doses per day
120 doses per container
0.15g tube
► Estimate
AET: (Tox Assessment Value)
Convert SCT (0.15 μgTDI) to μg/container
0.15 μg/day X 120 doses/container= 4.5 μg/tube
4 doses/day
4.5μg/tube
0.15g tube
=
30μg/g
How are unknowns measured?
► Estimated
►Based
on Response of
 Internal Standard
 Significant Identified Extractable Peak
► Final
 Incorporate Uncertainty
►RRF
Data Base
►1%RSD or 50% of Estimated AET
Uncertainty Factor
► Final
AET
Tube (30 ug/g)
50% correction of the estimated AET = 15ug/g
Estimated AET corrected for 1%RSD/35% =20ug/g
What does that mean?
Extractables ≥ The Estimated AET should be
Identified to the Extent Possible
Location of the AET (Extractables)
► Extraction
 2 grams (100 cm2) extracted in 100mL of 2-propanol
► AET
 AET response typical of UV antioxidant species
► Final
AET
 50% Factor
DA D1 A , Sig=200,4 Ref =550,100 (061838\SEP05029.D)
mA U
120
100
Estimated AET
Final AET
80
30ug/g
15/ug/g
60
40
20
0
0
2
4
6
8
10
12
14
16
min
#9 Polyaromatic Hydrocarbons
(PAH’s; or Polynuclear Aromatics,
PNA’s), N-nitrosamines, and
2-mercaptobenzothiazole (MBT) are
considered to be “special case”
compounds, requiring evaluation by
specific analytical techniques and
technology defined threshold
PAHs/PNAs
N-nitrosamines
Naphthalene
Chrysene
N-nitrosodimethylamine
Acenaphthylene
Benzo(b)fluoranthene
N-nitrosodiethylamine
Acenaphthene
Benzo(k)fluoranthene
N-nitrosodi-n-butylamine
Fluorene
Benzo(e)pyrene
N-nitrosomorpholine
Phenanthrene
Benzo(a)pyrene
N-nitrosopiperidine
Anthracene
Indeno(123-cd)pyrene
N-nitrosopyrrolidine
Fluoranthene
Dibenzo(ah)anthracene
Pyrene
Benzo(ghi)perylene
Benzo(a)anthracene
#10 Qualitative and quantitative
extractables profiles should be
discussed with and reviewed by
pharmaceutical development team
toxicologists so that any potential
safety concerns regarding
individual extractables, i.e.
potential leachables, are identified
early in the pharmaceutical
development process
Component Profiles
Qualitative
► Sampling and Preparation
► Multiple Solvents
► Sample to Surface Ratio
► Guided by the AET
Quantitative
► Evaluate Qualitative Profiles
► Optimize Method
 Extraction
► Asymptotic
 Analysis Conditions/Calibrations
 Method Accuracy/Precision
 Standard Reference
Materials
Multiple Extraction
Techniques
► Multiple Analytical
Techniques
►
 System Suitabilities
Levels
►
Uncertainty
 Finalize AET
►
Correlation
 Supplier Information
 Leachable Studies
Controlled Extraction Study
Summary of Steps
►
►
Qualitative Profile
Determine Extractable AET
 Convert SCT total daily intake to drug product relative units then
associate to the mass of the component
►
Quantitative Profile




►
Selection of Analytes
Asymptotic Extractions
Linear Dynamic Range
Consider Special Case Compounds
Optimize Method and Determine Range and Limits
 Recovery/Repeatability
 Based on Techniques used in Controlled Extraction Study
►
►
Validate Methods
What Next?
Routine Extractable Testing
► Test
Multiple Component Lots
► Correlate to Leachables
► Establish Specification and Acceptance
Criteria
► Component Control
Science Based Approach
►
►
►
►
►
►
Define expectation early
Understand and apply the science involved
Select appropriate CCS materials and components
Apply appropriate upstream controls
Communicate and Collaborate starting in early stages of
drug development
 With-In company
 With suppliers
 With regulatory bodies
PQRI WG proposal seems to support similar approach
Guirag Poochikian PhD.
PQRI Safety Thresholds and Best Practices
for Extractables and Leachables in OINDP
November 2005
Summary of PQRI Recommendations
►
Controlled Extraction Studies should:
 employ vigorous extraction with multiple solvents
 incorporate multiple extraction techniques
 include careful sample preparation based on knowledge of
analytical techniques to be used
 employ multiple analytical techniques
 define a systematic process for identification of individual
extractables
 optimize definitive extraction techniques/methods
 be evaluated relative to supplier information describing
formulation
 consider special case (PNA, MBT and nitrosoamines) separately
 review profiles with development team toxicologists to be
alerted to safety concerns regarding individual extractables
Conclusion
Extraction techniques/methods used for Controlled
Extraction Studies must be technically justified and
optimized to produced extractable profiles at least
equivalent to leachable profiles obtained under worst case
conditions of drug product use, allowing both qualitative
and quantitative extractable leachable correlations
► Properly conducted Controlled Extraction Studies, when
accomplished early in the pharmaceutical development
process, permit a pharmaceutical development team to
begin early evaluation of potential drug product leachables.
This evaluation can alert the pharmaceutical development
team to potential leachables with toxicological concern,
allowing adequate time to begin appropriate safety
qualification studies, or modification of CCS system.
►
Conclusion
► The
Best Practices recommendations for
Controlled Extraction studies are not meant to be
prescriptive or to exclude other scientifically valid
approaches, the analytical techniques/methods, or
control strategies
► These
recommendations represent a consensus
with-in the Working Group on current best
practices with-in the pharmaceutical industry and
are designed to reduce the level of uncertainty
with-in the pharmaceutical development process
for OINDP
References
Quality Systems Approach to Pharmaceutical Current Good Manufacturing
Practice Regulation; Draft Guidance for Industry US Department of Health and
Human Services FDA CDER/CBER/CVM/ORA, September, 2004
Metered Dose Inhaler (MDI) and Dry Powder Inhaler (DPI) Drug Products:
Chemistry Manufacturing and Control Documentation; Draft Guidance for
Industry; US Department of Health and Human Services Food and Drug
Administration CDER; October 1998
Nasal Spray Inhalation Solution, Suspension, and Spray Drug Products:
Chemistry Manufacturing and Control Documentation, Guidance for Industry;
US Department of Health and Human Services FDA CDER; July 2002,
Draft Safety Thresholds and Best Practices for Extractables and Leachables in
Orally Inhaled and Nasal Drug Products; and Experimental Protocols for
Controlled Extraction Studies; PQRI Leachables and Extractables Working Group;
Submitted July, 2006
References
►
PQRI Leachable Extractable Work Shop Presentations;
December 2005, www.pqri.org
 Schroeder, A.; Leachables and Extractables in OINDP: An FDA
Perspective
 Poochikian,G.; Best Practice Recommendations: Science and Process
 Ball, D; Derivation and Justification of Safety Thresholds
 McGovern,T.; Safety Recommendations: Science and Process
 Feinburg,T.; Controlled Extraction Studies
 Norwood,D.; Development and Application of the Analytical Evaluation
Threshold
 Paskiet,D.; Leachable Studies and Routine Extraction Studies
 Winkle,H.N.; Direction for Leachables and Extractables
References
ASTM Designation: D 5524-94 “Standard Test Method for Determination of
Phenolic Antioxidants in high Density Polyethylene Using liquid
Chromatography”
J.D. Vargo and K.L. Olson “Characterization of Addtives in Plastics by Liquid
Chromatography-Mass Spectrometry,” J. Chrom., pp.215-224 (1986)
Jenke, DR. Nomenclature Associated with Chmeical Characterization of and
Compatibility Evaluations of Medical Product Delivery Systems. PDA J Pharm
Sci Technology, 57 (2), pp. 97-108, 2003