PODP Approach to Acquire Extractable Profile Data - PQRIpqri.org/wp-content/uploads/2015/11/Egert.Hendricker.Houston.PODP... · PODP Approach to Acquire Extractable Profile Data Thomas

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




Part 1: The Experimental Protocol

Thomas EgertResearch Scientist

Boehringer Ingelheim Pharma GmbH & Co.KG



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



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 !


Summary of Results




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


Summary of Results


5



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


Summary of Results


Presenter

Presentation Notes



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:


Summary of Results


Presenter

Presentation Notes



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):


Summary of Results


Presenter

Presentation Notes



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)


Summary of Results


Presenter

Presentation Notes



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



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


Summary of Results


Presenter

Presentation Notes



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.

Presenter

Presentation Notes



13

Thermal n‐

HexaneIso‐

propanolIsopropan‐ol/Water

Aqueous pH 2.5

Aqueous pH 9.5


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

Presenter

Presentation Notes



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 . . .?


Summary of Results


Presenter

Presentation Notes



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)


Summary of Results


Presenter

Presentation Notes



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

Presenter

Presentation Notes



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


Summary of Results


Presenter

Presentation Notes



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.


Summary of Results


Presenter

Presentation Notes



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


Summary of Results


Presenter

Presentation Notes



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


min0 2 4 6 8 10 12 14 160

20000


min0 2 4 6 8 10 12 14 160

20000


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


Summary of Results


Presenter

Presentation Notes



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


Summary of Results


Presenter

Presentation Notes



22

Experi-mental Workflow

Presenter

Presentation Notes



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


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

Presenter

Presentation Notes



24

Phase 2/3: Sample PreparationExperimental Workflow

Presenter

Presentation Notes



25

Phase 3/3: Instrumental AnalysisExperimental Workflow

Presenter

Presentation Notes



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


Summary of Results


Presenter

Presentation Notes




Part 2: Summary of Results

Alan HendrickerCatalent Pharma Solutions





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


Summary of Results




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


Summary of Results


29



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)


Summary of Results


30



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


Summary of Results


31



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


Summary of Results


32



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


Summary of Results


33



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?


Summary of Results


34



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?


Summary of Results


35



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?


Summary of Results


36



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…


Summary of Results


Phthalic anhydride

3 µg/g

GC/MS – Chromatogram: Isopropanol (Reflux) extract of PVC



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).


Summary of Results


38




Part 3: Recommended Best Practices for Extractables Testing

Christopher T HoustonPrincipal Scientist, Bausch + Lomb




Summary of Results




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


Summary of Results




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

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Summary of Results




“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


Summary of Results




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


Summary of Results


Presenter

Presentation Notes

Source: Jenke LDPE report (sonication), Egert Ingleheim report (IPA/w sealed vessel)




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


Summary of Results


Presenter

Presentation Notes

Source: Jenke LDPE report (sonication), Egert Ingleheim report (IPA/w sealed vessel)




• 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


Summary of Results




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


Summary of Results




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


Summary of Results




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


Summary of Results




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


Summary of Results




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


Summary of Results


Presenter

Presentation Notes

Source: Jenke report on Polycarbonate



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


Summary of Results


Presenter

Presentation Notes

Source: Jenke PC report



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

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Summary of Results




“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


Summary of Results




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


Summary of Results


Presenter

Presentation Notes

Source: Polycarbonate report



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


Summary of Results




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


Summary of Results




“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


Summary of Results




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


Summary of Results


Presenter

Presentation Notes

Source: Experience from B+L method development effort on HPLC



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


Summary of Results




Example 9:100x Extract Concentration

GC-MS, total ion chromatograms60


Summary of Results


Presenter

Presentation Notes

Source: LDPE report



“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


Summary of Results




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


Summary of Results


Presenter

Presentation Notes

Source: Egert reports, Inglehein and Ridgefield data



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


Summary of Results




“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


Summary of Results




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


Summary of Results


Presenter

Presentation Notes

Data source: Primary BI Ridgefield and Ingleheim reports, Jenke COC report



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)



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)


Summary of Results


Epoxidized soybean oil from PVC



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


Summary of Results


Documents

PODP Approach to Acquire Extractable Profile Data - PQRIpqri.org/wp-content/uploads/2015/11/Egert.Hendricker.Houston.PODP... · PODP Approach to Acquire Extractable Profile Data Thomas