podp approach to acquire extractable profile data -...
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PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
PODP Approach to Acquire Extractable Profile Data
Thomas Egert Alan HendrickerChris Houston
Thresholds and Best Practices forParenteral and Ophthalmic Drug
Products (PODP)February 22-23, 2011
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
PODP Approach to Acquire Extractable Profile Data
Part 1: The Experimental Protocol
Thomas EgertResearch Scientist
Boehringer Ingelheim Pharma GmbH & Co.KG
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
3
PODP Working Plan Hypothesis
• Threshold concepts that have been developed for safety qualification of leachables in OINDP can be extrapolated to the evaluation and safety qualification of leachables in PODP, with consideration of factors and parameters such as dose, duration, patient population and additional product dependent characteristics unique to various PODP types.
• The “good science” best demonstrated practices that were established for the OINDP pharmaceutical development process can be extrapolated to container closure systemsfor PODP.
• Threshold and best practices concepts can be integrated into a comprehensive process for characterizing container closure systems with respect to leachable substances and their associated impact on PODP safety.
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
4
Work Plan Outline
• “. . . In order to test the hypothesis that best demonstrated practices for the characterization and analytical evaluation (of PODP) exist, the Working Group must establish such practices …“
Motivation for a Experimental Program (Phase I):
Controlled Extraction Studies !
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Safety Thresholds and Best Practices for Extractables and Leachables in Orally Inhaled and Nasal Drug Products 2006
Coming from OINDP Best Practices:Controlled Extraction Studies are the first
experimental milestone . . .Select components and/or raw
materials
Conduct rusk assessment oninformation from supplier
Individualingredient poses
unacceptablerisk
YES
Conduct controlled extractionstudies on components
Develop and validateextraction studies on
components
Establish correlation betweenleachables and extractables
profiles
Establish acceptance criteriafor leachables and extractables
Conduct leachables studies ondrug product and placebo
NO
Individualextractable greaterthan or equal to the
AET/SCT
NO
No further safety assessment
Individualextractable greaterthan or equal to the
AET/SCT
Report leachable totoxicologist for risk
assessment
Report extractable totoxicologist for risk
assessment
Go to safety qualificationprocess
NO
YES
YES
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
5
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
6
Recapitulation:What is the Purpose of a Controlled
Extraction Study
Verify and complement supplier information aboutmaterial
Establish a basis for the development and validation of routine quality control methods and acceptance criteria for critical componentsextractables profiles (consistency in composition)
Establish a basis for the development and validation of leachables methods
Allow for correlation of extractables and leachables
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
7
Nature of typical PODP materials and their(unique?) interactions with typical PODP drugproduct formulations (implications forextraction solvents and – techniques)
Universe of substances encountered
Applicability of AET – related concept (OINDP-paradigm) to extraction studies on PODP materials
Investigation of Workplan HypothesisImplies to Investigate:
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
8
Analytical methods:
Appropriate range of instrumental techniques( Likelihood of comprehensive evaluation of extractables)
Identification performance, suitable to generatedata for safety assessment based on:
(i) SAR endpoints(ii) confirmed identification
Sensitivity (in terms of lowest level foridentification)Specificity (matrix interferences) Limitations (critical substances/mixtures)?
Investigation of Workplan HypothesisImplies to Investigate (cont‘d):
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
9
PQRI PODP Experimental ProtocolThe General Concept
Test Articles representing PODP materialsAppropriate extraction techniquesAppropriate solventsVarious analytical techniquesVarious participating laboratories(experienced in the field of E&L)Comprehensive and detailed experimental protocolSemiquantitative approach –Reporting Limit 10 µg/gQuality requirements (system suitability)
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
10
Test Articles Representing PODP Materials
Test Articles (Material Type)
Format Composition (Supplier Information)
Application Category
Polycarbonate (PC)
Injection moulded plaques
• 0.05 PHR Irganox 1076 • 0.1 PHR Irgafos 168
Ports, Tubes
LVP
Rubber Elastomer (Bromobutyl)
Sheet • Brominated isobutylene isoprene copolymer (57.3%)
• calcined aluminum silicate, 38.2% • titanium dioxide, 1.2%; • paraffinic oil, 1.2%; • zinc oxide, 0.6% • polyethylene0.6% • SRF Carbon block mixture, 0.4% • calcined magnesium oxide, 0.3% • 4,4’-dithiodi-
morpholine/polyisobutylene, 0.3%
Closures, Plungers, Gaskets
SVP
Cyclic Olefin Copolymer (COC)
Plaques
• Irganox 1010 • Ultramarine Blue
Syringes, Vials
PFS, SVP
Polyvinylchloride (PVC)
Pellets • PVC resin • DEHP 30% • Epoxidized oil 7% • Zn stearate 0.5% • Ca stearate 0.5% • Stearamide 1%
Bags, Tubing
LVP
Low density polyethylene (LDPE)
Blown Film • Irganox B 215 (2:1 blend of Irgafos 168 and Irganox 1010) 1000 ppm
• BHT 200 ppm • Calcium Stearate 500 ppm • Erucamide 500 ppm • Chimassorb 944 2000 ppm
Overpouch, BFS, Containers
BFS, SVP, LVP
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
11
Solvents should mimic drug product formulation
The majority of PODP are represented by aqueous basedformulations !Cosolvents can be subdivided into two groups:
A: Polarity Neutral
Primary function of excipient is not drugsolubilizationGenerally compounds with high aqueous solubility:
Diluents (dextrose, saline)Buffers (acetate, lactate, bicarbonate, phosphate)Amino acidsVitamins
B: Polarity Impacting
Components primary function is to increase the solubility of the drug
Tween 80CyclodextrinsSDSLipids up to 20% wt/wtSurfactants, Emollients
Isopropanol / WaterAqueous pH 2.5 / 9.5
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
12
Extraction/Solvent Map
Thermal n‐
HexaneIso‐
propanolIsopropan‐ol/Water
Aqueous pH 2.5
Aqueous pH 9.5
Headspace X ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ Reflux ‐‐‐ X X PC/PVC only ‐‐‐ ‐‐‐ Soxhlet ‐‐‐ X X ‐‐‐ ‐‐‐ ‐‐
Sealed Vessel ‐‐‐ ‐‐‐ ‐‐‐ 55°C/3d (121°C/1hr)1
(121°C/1hr)1
Sonication ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ x x 1: autoclave conditions: (121°C/1hr)
Solvent Polarity /Drug Product Similarity
Tem
p.
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
13
Thermal n‐
HexaneIso‐
propanolIsopropan‐ol/Water
Aqueous pH 2.5
Aqueous pH 9.5
Headspace X ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ Reflux ‐‐‐ X X PC/PVC only ‐‐‐ ‐‐‐ Soxhlet ‐‐‐ X X ‐‐‐ ‐‐‐ ‐‐
Sealed Vessel ‐‐‐ ‐‐‐ ‐‐‐ 55°C/3d (121°C/1hr)1
(121°C/1hr)1
Sonication ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ x x 1: autoclave conditions: (121°C/1hr)
Extraction “Paradigms*” in Laboratory Practice
• Vigorous/exaggerated/-exhaustive conditions
• no material deformulation„Hot“ extraction techniques but no sample dissolving solvents
• Solvents should be attributed to the expected universe of substances (cover wide range of polarity)
• Solvents should mimic drug product formulation
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
14
Analytical Methods – General AspectsPrimary Focus:
Specific analytical procedures
(non specific could have supplemental character (e.g. gravimetry, photometry, total organic carbon, alkalinity, acidity, reducing subtances, infrared, thermal gravimetry etc.) pharmacopoeial testing
Non-Target Analysis in addition to targets known from
composition
“Small“ molecules (< 1000 Da)
Trace (Organic) Analysis
Standard – chromatographic conditions suitable to
efficiently separate the majority of the log Po/w range to
be expected.
What you might been missing . . .?
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
15
Analytical Methods…choosing the most adequate tools …
InorganicTrace elements and metals
ICP/MS
OrganicVolatiles substances:
Static Headspace-GC/MS
Semivolatiles (GC-amenable)GC/MS
Semivolatiles (not GC-amenable)LC/UVLC/MSn, HRMS
Philosophy: Identification to an extent practicable . . .
(OINDP Best Practices)
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
16
Conditions for Analytical Methods(Chromatography)
Headspace-GC/MS
Column: Oven-Program DB WAXETR
60 m x iD 0.32 mm, FT 1.0 µm He 5 psi
35°C-7 min-1 K/min - 40°C – 15 min - 10 K/min – 100 - 25 K/min – 240 - 5 min MS: EI 70 eV, 25 – 200 amu Headspace-Cond.: 80°C - 120 min
GC/FID (MS) Column: Oven-Program DB 5HT
30m x 0.25 mm, FT 0.25µm 50°C – 5 min – 10 K/min – 330°C – 5min Inj.-Vol. 1 µl, splitless Injector: 310 °C FID: 150°C MS: EI 70 eV, 33-650 amu
LC/UV (MS)
Column: Mobile-Phase Agilent Zorbax Exclipse Plus
C18, 100 x 3.0 mm, 3.5 µm particles Column Oven : 40 °C
A : 10 mM ammonium acetate B : acetonitrile Flow rate: 0.8 mL/min Gradient:
Sample Size: 10 µl Time: %B 0.0 5.0 Detection 8.4 100.0 UV 205 - 300 nm 35.0 100.0 MS API-ES positive and
negative ion (mass range 80 - 1200)
36.0 5.0 39.0 5.0
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
17
Identification to “the Extent Practicable”…Identification attributes according to OINDP Best Practices have been applied
Category Supporting Identification Data
A Mass spectrometric fragmentation behaviour
B Confirmation of molecular weight
C Confirmation of elemental composition
D Mass spectrum matches automated library orliterature spectrum
E Mass spectrum and chromatographic retentionindex match authentic specimen
Confimed Categories A, B(or)C and D(or)E fulfilled
Confident Sufficient Data to preclude all but the mostclosely related structures have been obtained
Tentative Data have obtained that are consistent with a class of molecule only
Confirmed ID
SAREnd-points
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
18
System Suitability Requirements were applied to monitor result integrity
Level 1 - Qualified Instrumentation
Proper instrument condition and instrument suitabilty will be demonstrated by each participating laboratory by following its proprietay (inhouse) procedures
Level 2 – System suitability mixtures
Specific test mixtures to be analyzed by HS-GC, GC, LC and ICPTest mixtures are suitable to demonstrate adequate and effective analytical
performance (separation efficiency, selectivity, sensitivity)Results be evaluated against defined acceptance criteria
Level 3 - Internal standardization
Surrogate Internal Standard – added to the extract control of effectiveness of the sample preparation process
Injection Internal Standard – added to the injection solution control of the sample introduction and chromatographic process for each
sample run.
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
19
Suitability Mixtures HS-GC and GCHS-GC
Custom-Mix: (µg/vial)
methanol 2
10.00 15.00 20.00 25.00 30.00 35.00 40.00
1000000
2000000
3000000
4000000
5000000
6000000
7000000
8000000
9000000
1e+07
1.1e+07
1.2e+07
1.3e+07
1.4e+07
1.5e+07
1.6e+07
1.7e+07
1.8e+07
1.9e+07
2e+07
>
dance
TIC: 9NOV2009003.D\data.ms
16.608
31.309
32.01433.201
37.957
39.064
39.182
acetic acid 2 cyclohexanone 1 toluene 1 trimethylsilanol 2 2-ethyl hexanol 2
GC “Grob”-Mix: (µg/ml) L(+)-2,3-butanediol 27
8 9 10 11 12 13 14 15
100000
200000
300000
400000
500000
600000
700000
800000
900000
1000000
1100000
1200000
1300000
Time-->
Signal: 1201014.D\FID1A.CH
Grob Mix (1/100)
1
2
4
3
5
67
8
9
10
11
12
n-decane 14 2,6-dimethylaniline 16 2,6-dimethylphenol 16 methyl decanoate (C10:0) 21 methyl docecanoate (C12:0) 21 methyl undecanoate (C11:0) 21 nonanal 20 1-octanal 18 n-undecane (C11) 14
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
20
Suitability Mixtures LC and ICPLC (UV/MS) Custom-Mix: (µg/ml)
min0 2 4 6 8 10 12 14 16
mAU-500
DAD1 A, Sig=210,20 Ref =360,100 (JIJ2009\JI000003.D)
min0 2 4 6 8 10 12 14 16
50000
MSD1 TIC, MS File (JIJ2009\JI000003.D) API-ES, Neg, Scan, Frag: 100, "Negativ e"
min0 2 4 6 8 10 12 14 160
200000
MSD2 TIC, MS File (JIJ2009\JI000003.D) API-ES, Pos, Scan, Frag: 100, "Positiv e"
min0 2 4 6 8 10 12 14 160
MSD2 114, EIC=113.7:114.7 (JIJ2009\JI000003.D) API-ES, Pos, Scan, Frag: 100, "Positiv e"
min0 2 4 6 8 10 12 14 160
MSD1 227, EIC=226.7:227.7 (JIJ2009\JI000003.D) API-ES, Neg, Scan, Frag: 100, "Negativ e"
min0 2 4 6 8 10 12 14 160
MSD1 277, EIC=276.7:277.7 (JIJ2009\JI000003.D) API-ES, Neg, Scan, Frag: 100, "Negativ e"
min0 2 4 6 8 10 12 14 160
20000
MSD1 299, EIC=298.7:299.7 (JIJ2009\JI000003.D) API-ES, Neg, Scan, Frag: 100, "Negativ e"
min0 2 4 6 8 10 12 14 160
20000
MSD1 283, EIC=282.7:283.7 (JIJ2009\JI000003.D) API-ES, Neg, Scan, Frag: 100, "Negativ e"
min0 2 4 6 8 10 12 14 160
100000
MSD2 391, EIC=390.7:391.7 (JIJ2009\JI000003.D) API-ES, Pos, Scan, Frag: 100, "Positiv e"
caprolactam 1 butylatedhydroxytoluene 5
diphenylamine 5 mono-(2-ethylhexyl)
phthalate 1
stearic acid 5 di-(2-ethylhexyl phthalate) 1 bisphenol A 1
ICP
all target elements
0.25
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
21
Internal StandardsSurrogate Internal Standard Objective Requirements Substance used Monitoring of sample preparation process and instrumental performance
sufficiently stable sufficiently soluble in all extraction
solvents amenable to back-extraction from
aqueous extracts by organic solvents amenable to TMS-derivatization semi-volatile amenable to all detection principles selectively detectable
Bisphenol M CAS 13595-25-0 MW: 346.46
Injection Internal Standard Objective Requirements Substance used Monitoring of instrumental performance
sufficiently stable sufficiently soluble in final extract semi-volatile amenable to all detection principles selectively detectable
Irganox 415 CAS: 96-69-5 MW: 358.538
Headspace Internal Standard
1,4-Dioxane CAS: 123-91-1 MW: 88.11
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
22
Experi-mental Workflow
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
23
Aqueous ExtractpH 2.5200 ml
Aqueous ExtractpH 9.5200 ml
IPA/Water extract200 ml
Extraction Techniques:Soxhlet (min. 10 cycles, 24 hrs, 5 g- 200 ml)Reflux (2 hrs, 5 g - 200 ml)Sonication (2 hrs, T=0°C, 5 g - 200 ml)Sealed Vessel (55 °C / 3 d, 5 g- 200 ml)Sealed Vessel Autoclaved(121 °C / 1 hr 5 g - 200 ml, 2 replicates)
IPA extract200 ml
N-Hexane extract200 ml
Test Articles:
Sample Weight [5g]Sample Pre-treatment
Materials:LDPEPCPVCCOCRubber
HS-GC/MS(FID)
ICP/MS(AES)
Phase 1/3: Extract Preparation
Thermal n‐
HexaneIso‐
propanol Isopropan‐ol/Water
Aqueous pH 2.5
Aqueous pH 9.5
Headspace X ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ Reflux ‐‐‐ X X PC/PVC only ‐‐‐ ‐‐‐ Soxhlet ‐‐‐ X X ‐‐‐ ‐‐‐ ‐‐
Sealed Vessel ‐‐‐ ‐‐‐ ‐‐‐ 55°C/3d (121°C/ 1hr)2
(121°C/ 1hr)2
Sonication ‐‐‐ ‐‐‐ ‐‐‐ ‐‐‐ x x 1: All test articles (materials) were extracted following this scheme if not indicated otherwise 2: autoclave conditions: (121°C/1hr)
Experimental Workflow
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
24
Phase 2/3: Sample PreparationExperimental Workflow
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
25
Phase 3/3: Instrumental AnalysisExperimental Workflow
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
26
Challenges Ahead:What could have been missed?
Example: Epoxidized soybean oil (specif. PVC stabilizer):
Mixture of glycerol esters –75 possible structuresconsisting of: (~ %)
Palmitic acid C16:0 11Stearic acid C18:0 5Epoxidized oleic acid C18:1 8
Epoxidized linoleic acid C18:2 23
Epoxidized linolenic acid C18:3 54
Identified and quantified ?Almost missed ?Completely missed ?
Epoxidized trilinoleinC57H98O12, MW 975.45 g/mol
O
OO
O
O
O
O
O
O
O
O
O
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
PODP Approach to Acquire Extractable Profile Data
Part 2: Summary of Results
Alan HendrickerCatalent Pharma Solutions
Thresholds and Best Practices forParenteral and Ophthalmic Drug
Products (PODP)February 22-23, 2011
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Test Articles.
Material Type Material Application
Material Format
Composition
Low density polyethylene (LDPE)
Overpouch Blown Film Dow 640-I LDPE resin; Irganox B 215 (2:1 blend of Irgafos 168 and Irganox 1010) 1000 ppm, BHT 200 ppm, Calcium Stearate 500 ppm, Erucamide 500 ppm, Chimassorb 944 2000 ppm
Cyclic Olefin (COC) Syringe barrels, vials Plaques Irganox 1010, Ultramarine Blue
Polycarbonate (PC) Port Tubes Injection molded plaques
0.05 Parts per Hundred (PHR) Irganox 1076, 0.1 PHR
Irgafos 168
Poly (vinyl chloride) (PVC)
Solution Bags, tubing
Pellets PVC resin; DEHP 30%; Epoxidized oil 7%, Zn
stearate 0.5%; Ca stearate 0.5%; Stearamide 1%
Rubber (Elastomer) (RE) Gaskets, stoppers, closures
Sheets Brominated isobutylene isoprene copolymer (57.3%); calcined
aluminum silicate, 38.2%, titanium dioxide, 1.2%; paraffinic oil, 1.2%;
zinc oxide, 0.6%; polyethylene, 0.6%; SRF Carbon block mixture, 0.4%; calcined magnesium oxide,
0.3%; 4,4’-dithiodi-morpholine/polyisobutylene, 0.3%
Test Articles for Extractables Studies
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Poly(vinyl chloride) PVC Test Material
Metals Results all less than 5 µg/g. Calcium and Zinc(known additives) detected in most extracts. MaximumCalcium 1.72 µg/g, Zinc 1.33 µg/g
Volatiles testing (HS-GC-MS) showed primarily aliphatichydrocarbons of short chain length, branched andunbranched. One volatile identified was attributed to 2-ethyl-1-hexanol, a known extractable from PVC.
Semivolatiles testing (GC-MS) showed a large number ofpeaks (up to 50) for each extraction condition. Peaks wereattributed to fatty acids, fatty amides, fatty alcohols, fattyacid esters, fatty aldehydes, aliphatic hydrocarbons, BHT,phthalates, phthalate esters and phthalate degradationproducts. Fatty acids and related peaks were observed atthe highest concentrations (up to around 500 ppm).
Non-volatiles testing (LC-MS) confirmed GC-MS results,peaks detected were attributed to fatty acids, fatty amides(erucamide, (z)-9-Octadecenamide), and phthalates.
Example of a PVC IV Bag
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
29
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Polycarbonate (PC) Test Material
Metals results showed only trace level metals except in oneextraction condition (Autoclave/pH 2.5) which showedapproximately 50 µg/g of Zn and 25 µg/g of Ca.
Volatiles testing using HS-GC-MS showed only low levels of a fewvolatiles with a maximum concentration of 0.12 µg/g for nonanal.Acetone and acetonitrile were also observed at trace levels.
Semivolatiles testing of extracts using GC-MS showed asignificant number of peaks, including several phenolic peakswhich are likely degradation products of the polymer itself. Mostnotable of these was Bisphenol-A, a compound of significantpotential toxicity. It is not known the extent to which this speciesis generated via the extraction versus being present atendogenous levels. Other species detected included Irgafos 168and its oxidation product, Irganox 1076, 2,4-di-t-butylphenol anddimethyphthalate.
Non-volatiles testing (LC-MS) showed a significant number ofpeaks, many of which overlapped and for which first passidentification was not possible. They may be various oligomericfragments related to the base polymer.
Example of a Polycarbonate Baby Bottle (no
longer marketed)
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
30
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Cyclic Olefin Copolymer (COC) Test Material
Metals results showed only trace levels of metals in general.One extraction condition (Autoclave/pH 2.5) showedapproximately 25 µg/g of Mg 44 µg/g of Zn and 68 µg/g ofBromine.
Volatiles testing using HS-GC-MS showed very cleanprofiles, only one peak attributed to Cis-decahydro-naphthalene was observed at 0.03 µg/g.
Semivolatiles testing of extracts using GC-MS showed alarge number of peaks, especially via sealed vesselextraction. Peaks detected were attributed to fatty acids,fatty acid esters, siloxanes, phthalate related peaks and asignificant number of peaks which could not be given firstpass identifications. The intensity of peaks observed in allextracts was very small, typically less than 0.1 µg/g. Withseveral extraction technique, no discernable extractablescould be detected using this method.
Non-volatiles testing (LC-MS) showed no peaks in sealedvessel or sonication extractions. Reflux extraction in IPAproduced several low level peaks which could not beidentified (less than approximately 0.1 µg/g).
COC Syringe
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
31
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Low density polyethylene (LDPE)Test Material
Metals testing results showed no significant metalextractables (all less than 1 µg/g). Calcium (knownadditive) was not detected under any extractioncondition.
Volatiles testing using HS-GC-MS showed only one peakabove the AET, which was late eluting and could not beidentified. It was present at 0.26 µg/g.
Semivolatiles testing of extracts using GC-MS showedBHT (antioxidant), Z-9-octadecenamide (antistatic)erucamide (antistatic) and Dilauryl-3,3'-thiodipropionateor Irganox PS 800 (heat stabilizer and antioxidant). Anumber of aliphatic hydrocarbons branched andunbranched can be observed, oligomeric fragments ofthe PE itself.
Non-volatiles testing (LC-MS) showed no additionalidentifiable extractables for the tests performed, thoughseveral peaks were observed at 220nm.
Polyethylene resin beads
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
32
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Rubber Elastomer (RE) Test Material
Metals results showed significant amounts of Bromine, up to 20µg/g, which was not surprising, since it was a bromobutylelastomer. Smaller amounts (all less than 10 µg/g) were detectedof the other known metal additives, including Mg, Al, Ca, Ti andZn. The pH 2.5 sonication extraction showed the highest levels ofthese metal species.
Volatiles testing using HS-GC-MS showed several peaksattributed to butyl oligomers. Peaks attributed to methylcyclopentane and cyclohexane were also observed. All peaksexcept cyclohexane were less than 1 µg/g, exceptmethylcyclopentane at 1.19 µg/g.
Semivolatiles testing of extracts using GC-MS showed anextremely complex extractables profile, with large numbers ofunknowns. Peaks identified were attributed to fatty acids, fattyacid esters, fatty amides, aliphatic hydrocarbon species, BHT, andbrominated oligomers.
Non-volatiles testing (LC-MS) showed a complex unresolvedenvelope of dozens of peaks, especially for organic extracts. Themethod utilized was insufficient for peak identifications in mostcases.
Example of Elastomeric Stoppers
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
33
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Observations
Profiles can be complex! Withmultiple extraction solvents andtechniques hundreds of peaks can beextracted using the chromatographicmethods
AET should be employed to helpsimplify data analysis
In general, residual solvents andmetals testing showed low levels ofextractables and were generallyinnocuous species, but providedvaluable information See the forest
through the trees?
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
34
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Observations
Understand that the peaks you aredetecting are not necessarily what wasadded to the polymer originally (e.g. 2,4-di-t-butylphenol and palmitic acid canoriginate from Irgafos 168 and CalciumStearate, among others)
Use “realistic” extraction solvents tounderstand risk. Use aggressive solventsto facilitate identifications
Oligomeric fragments representchallenging species for completestructural identification. However,compound class may be enough toassess toxicological risk
6 8 10 12 14 16 18 20 22 24 26
15000
20000
25000
30000
35000
40000
45000
50000
55000
Time-->
AbundanceSignal: RS015.D\FID1A.CH
B - Internal Standard 2 (Bisphenol M)A - Internal Standard 1 (Irganox 415)
Underivatized PVC pH9.5 Samples Overlay B
A30
30
32
27
25 23 21 22
19 18 14 13 9
3
1
PVC-pH9.5-2
PVC-pH9.5-1
pH9.5-Blank-2
Signal: RS020.D\FID1A.CH (*)
Signal: RS021.D\FID1A.CH (*)
6 8 10 12 14 16 18 20 22 24 26100000
200000
300000
400000
500000
600000
700000
800000
900000
1000000
1100000
1200000
1300000
Time
Response_
Signal: RS006.D\FID1A.CH
x - Peaks with this symbol are similar in size, Extract vs Extraction blank
21
ISTD2
xRE-IW-2
RE-pH9.5-2
RE-pH2.5-1
38
ISTD1
36
32
29
2826
2826
25
24
2120
20
18
16
1817
1615
14
1111
10
1110
98
543
2
x
x
x
1
1
x
x
Signal: RS017.D\FID1A.CH (*)Signal: RS027.D\FID1A.CH (*)
UNDERIVATIZED RUBBER ELASTOMER SAMPLES OVERLAY
Do you see the trees now?
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
35
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Observations
Solvents pH may change extractables(specific example shown subsequently)
Use HPLC-MS to complement GC-MS. Asa standalone technique data interpretation inLC-MS can be difficult
In some cases we did not observe knownadditives. Understand you may need to alterextraction conditions in some cases tofacilitate extraction. Or realize that you maynever see them even through due diligence(e.g. consumed processing agent)
min5 10 15 20
mAU
-100
0
100
200
300
400
DAD1 A, Sig=220,4 Ref=550,50 (F:\3\500_B...SETS\50_RESULTS_MATERIALS\03_RE\LC-UV\013-1101_REF3_NHEX_RE.D)
min2.5 5 7.5 10 12.5 15 17.5 20 22.5
mAU
-100
0
100
200
300
400
DAD1 A, Sig=220,4 Ref=550,50 (F:\3\500_B...SETS\50_RESULTS_MATERIALS\03_RE\LC-UV\013-1101_REF3_NHEX_RE.D)
7.5
53 7
.752
8.6
60 9
.267
10.
149
10.
443
10.
899
11.
827
12.
124
12.
447
12.
755
13.
218
13.
370
13.
926
14.
472
15.
502
16.
617
17.
921
Where did my trees (peaks) go?
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
36
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
37
Observations
How low could we go?
• By applying state-of-the-art analytical procedures and instrumentation, the limits of identification for single chemical entities (extractables) observed were in the range of 0.1 – 100 µg/g of material…
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
Phthalic anhydride
3 µg/g
GC/MS – Chromatogram: Isopropanol (Reflux) extract of PVC
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Observations (and Last Tree Analogy)
Maroon Bells, Aspen Colorado
In the end, materials extractables characterization can produce a complex scene, difficult to interpret or understand but full of information and when done correctly, paints a great picture. See the posters for the full picture (warning: not quite as nice as the one below).
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
38
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
PODP Approach to Acquire Extractable Profile Data
Part 3: Recommended Best Practices for Extractables Testing
Christopher T HoustonPrincipal Scientist, Bausch + Lomb
Thresholds and Best Practices forParenteral and Ophthalmic Drug
Products (PODP)February 22-23, 2011
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Best Practices
• In September 2006, PQRI issued prior guidance for OINDP:
“Safety Thresholds and Best Practices for Extractables and Leachables in Orally Inhaled and Nasal Drug Products”Contained 10 best practices with respect to controlled extraction studiesMost of these are quite relevant to PODPPODP nuances result from aqueous drug product formulations
40
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Review of OINDP Best Practice Recommendations
1. Controlled Extraction Studies should employ vigorous extraction with multiple solvents of varying polarity
2. Controlled Extraction Studies should incorporate multiple extraction techniques3. Controlled Extraction Studies should include careful sample preparation based on
knowledge of analytical techniques to be used4. Controlled Extraction Studies should employ multiple analytical techniques5. Controlled Extraction Studies should include a defined and systematic process for
identification of individual extractables6. Controlled Extraction Study “definitive” extraction techniques/methods should be
optimized7. During the Controlled Extraction Study process, sponsors should revisit supplier
information describing component formulation8. Controlled Extraction Studies should be guided by an Analytical Evaluation
Threshold (AET) that is based on an accepted safety evaluation threshold9. Polyaromatic hydrocarbons, N-nitrosamines, and 2-mercaptobenzothiazole (MBT) are
considered to be “special case” compounds, requiring evaluation by specific analytical techniques and technology defined threshold
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
41
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
“Controlled Extraction Studies should employ vigorous extraction with multiple
solvents of varying polarity”
• Prior OINDP recommendation:Advocated use of water in extraction studies for aqueous productsDiscouraged using water as the sole extraction solvent for components from aqueous drug products
• PODP Experience:Aligned with prior recommendation (Examples 1 - 3)Relevance of pH (Examples 4 and 5)
42
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 1:LDPE / Aqueous Drug Product
• In PODP protocol, extracted LDPE with known additive package by multiple solvents / techniques including water
In ophthalmology, significant prior art for extractables used water as the only solvent
• Although aqueous solvent extracts may be useful for gauging significant extractables, they may not promote understanding of the material
43
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 1:LDPE / Aqueous Drug Product
KnownAdditive
Sonication pH 2.5
Sonication pH 9.5
Sealed Vessel
IPA/WaterIrganox 1010
Irgafos 168
BHT
Ca Stearate(as stearic acid)Erucamide
Red = Not detected / Green = detected by GC and/or HPLC 44
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 1:LDPE / Aqueous Drug Product
• Sonication in aqueous solvents successfully detects erucamide, but no other anticipated additive
• Solvents of different polarity provide better understanding of the material
• Relevant to an aqueous product?An aqueous extraction profile lacking extractables does not necessarily impart material knowledgeThough non-polar additives such as Irganox 1010 are less likely to migrate into aqueous product, its identification alerts the researcher to look for more polar degradation / transformation products
45
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 2:Extraction of DEHP from PVC
• PVC material contained 30% bis(2-ethylhexyl) phthalate (DEHP)
• Sealed vessel extracts:
pH 2.5
pH 9.5
IPA/Water
DEHP
46
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 2:Extraction of DEHP from PVC
• DEHP is a significant extractable from PVC• Aqueous (pH 2.5, 9.5) solvent detect
significantly less DEHP than IPA/water• Many “aqueous” formulations are not purely
aqueous, but contain surfactants or other excipients that enhance solubility
• Using water as the only extraction solvent runs the risk of not observing a key extractable that is poorly water soluble but nonetheless soluble in drug product
47
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 3:Solvent Polarity and Polycarbonate
• The strongest solvent does not always yield the worst-case extraction profile
• Consider this example of isopropanol and n-hexane extracts generated by Soxhlet and reflux
48
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 3:Solvent Polarity and Polycarbonate
ExtractableEstimated Quantity (μg/g)
IPA Reflux
IPA Soxhlet
Hexane Reflux
Hexane Soxhlet
4-t-butylphenol 65.7 13.7 2.2 2.92,4-di-t-butylphenol 56.0 24.3 2.0 29.4Bisphenol A 77.1 9.4 9.4 12.7Irgafos 168 35.9 13.6 0.3 0.8Irgafos 168, oxidized 25.1 17.2 0.0 1.6Irganox 1076 21.8 8.3 0.1 0.5
IPA is a superior solvent for many of theseextractables, despite being “weaker”
49
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 4:pH Effects and Polycarbonate
• Polycarbonate extraction at pH 2.5 and 9.5
pH 2.5
pH 9.5
IPA/Water
BPA
50
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 4:pH Effects and Polycarbonate
• Bisphenol A detected from polycarbonatepH 2.5: 0.12 mg/LpH 9.5: 1.7 mg/LNot detected in isopropanol/water extract
• At alkaline pH, an order of magnitude more BPA is observed than at acidic pH
• Implications for product pH and packaging compatibility
51
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 5:pH and Solvent Effects / PVC
• Diethylhexyl phthalate (DEHP) and monoethylhexyl phthalate (MEHP) detected
• HPLC semiquantitative results:
• DEHP and MEHP are extracted in significantly higher quantities at basic versus acidic pH
• No single solvent system is sufficient for all analytes
Extractable pH 2.5(mg/L)
pH 9.5(mg/L)
IPA/water(mg/L)
DEHP 0.05 0.79 930
MEHP 0.02 0.57 0.09
52
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
“Controlled Extraction Studies should incorporate multiple
extraction techniques”
• Prior OINDP recommendations:Justification needed for low temperature techniques such as sonication with respect to extraction efficiency
Sonication found to be less efficient than Soxhlet and reflux by OINDP working group
Extraction profiles from higher temperature extraction techniques should be carefully examined for extraction artifacts.
• PODP ExperienceAqueous extraction by sonication versus sealed vessel (Example 6)Thermal methods, headspace GC (Example 7)
53
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 6:Sonication v. Sealed Vessel / Polycarbonate
• Sealed vessel (121 °C, 1 hr) at pH 2.5 or 9.5Bisphenol A detected
0.12 mg/L @ pH 2.51.7 mg/L @ pH 9.5
• Sonication (2 hr, 0 °C) at pH 2.5 or 9.5No bisphenol A detected, even at high pH
• Sealed vessel extraction was more efficient than sonication in this example
• Sonication required significant effort to standardize temperature relative to sealed vessel
54
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 7:Thermal Methods / Headspace GC
• Headspace GC (80 °C, 2 hours) on all materials• Observed extractables generally included low levels
of hydrocarbonsMultiple hydrocarbons for PVC ranging from 0.5 to 4.6 μg/gButyl rubber related oligomers in the rubber/elastomer sample
• Low parity with other extraction approaches, including Soxhlet and reflux with IPA or hexane
• However…
55
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 7:Thermal Methods / Headspace GC
• Headspace GC holds significant value in some PODP dosage forms
• Certain dosage forms in semipermeable packaging systems are susceptible to volatile leachables from secondary packaging systems
Examples to be provided in Ophthalmology session
• Stay tuned for further evaluation of headspace…
56
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
“Controlled Extraction Studies should include careful sample preparation based on knowledge of analytical
techniques to be used”
• Prior OINDP recommendations:Promoted the notion of solvent exchange when extraction solvent not suitable for use with analytical method
• PODP Experience:Solvent exchange (Example 8)Extract concentration, 100X (Example 9)
57
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 8:Solvent Exchange for HPLC
• The HPLC method developed for the PODP protocol intended to span a broad polarity.
• Caprolactam included in the system suitability mixture as the most polar compound
• With low retention, caprolactamexhibits poor peak shapeif injection solvent is too strong
• Underscores necessity forsolvent exchange from non-polar extracts
58
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 9:100x Extract Concentration
• Aqueous extracts were back-extracted into methylene chloride for GC analysis
• A 100x concentration step was performed to increase sensitivity
• In such concentration schemes, care must be taken to ensure that background contaminants are not concentrated with samples…
59
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 9:100x Extract Concentration
GC-MS, total ion chromatograms60
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
“Controlled Extraction Studies should employ multiple analytical
techniques”
• Prior OINDP recommendations:No single analytical technique can detect and identify all possible extractablesTechniques used should be “compound specific”Detector response proportional to extractable quantity
• PODP Experience:Aligned with prior recommendation (Example 10)
61
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 10:Extractables Detected in LDPE
Additives Detected by HPLC (UV,MS)
Detected by GC (FID,MS)
Irganox 1010
Irgafos 168
BHT
Stearic acid
Erucamide
Irganox 1010, related
Irgafos 168, related
DEHP*
Oleamide*
* Not included in resin composition information 62
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 10:Extractables Detected in LDPE
• Strength of multiple analytical survey techniques for organic extractables:
Although overlap exists, HPLC and GC contain some complementary dataWhere overlap exists, the different techniques provide confirmation and may aid in identificationMultiple survey methods allows for broader characterization, including the identification of unanticipated extractables (DEHP and oleamide in Example 9)
• Inorganic analysesThe PODP protocol also included ICP-MS for elemental analysisIf metals are a concern, the added orthogonality of atomic spectroscopy is critical
63
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
“During the Controlled Extraction Study process, sponsors should revisit supplier information describing
component formulation.”• Prior OINDP recommendations:
Check list of experimentally-derived extractables against supplier information
Are anticipated extractables observed?Are other extractables observed?
Supplier information can serve as a starting point for the development of analytical methods
• PODP Experience:Experimental work can reveal extractables that were not anticipated based on supplier information (Example 11)
64
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Example 11:Extraction Studies of COC
• Additives known to be formulated into the COC:Irganox 1010, ultramarine blue (pigment)
• Noteworthy examples of extractables observed during the PODP study of COC:
Irganox 1010, monoethylhexyl phthalate, DEHP, cis/trans-decahydronaphthalene, oleamide, hexadecanoic acid, octadecanoic acid
• Additional extractables observed from COC material beyond those anticipated from supplier information
• Sponsors cannot claim to understand critical component chemistry simply on the basis of supplier information
65
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Controlled Extraction Study “definitive” extraction techniques/methods should
be optimized
• Prior OINDP Recommendations:After first pass extraction studies, development team should choose a “definitive” method / technique to optimize.Complete validation is not recommended or expected for Controlled Extraction studiesMethod should be demonstrably fit for purpose with respect to accuracy and precision
• PODP Experience:Some extractables may require unique or dedicated methods to meet this objective (Example 12)
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
67
Example 12: “Borderline” Analytes (ESBO)
O
OO
O
O
O
O
O
O
O
O
O
Epoxidized trilinoleinC57H98O12, MW 975.45 g/mol
ESBO „Pattern“ in LC/MS( not quantifiable)
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing
Epoxidized soybean oil from PVC
PODP E&LWorking Group
T. Egert, A. Hendricker, C. HoustonBethesda, February 2011
Conclusion
• The PODP protocol underscores the value of extrapolating the OINDP best practice recommendations to other dosage forms
• For many PODPs, water is an essential solvent and suggests nuances on OINDP recommendations
Although the OINDP recommendations discuss water, that work focused on more non-polar solvents (isopropanol, hexane, methylene chloride)PODP work demonstrates the importance of pH as a consideration for extraction solvent selection
68
PODP Experimental Protocol
Summary of Results
Recommended Best Practices for Extractables Testing