Extractables and Leachables Testing: An Introduction...

www.intertek.com1

Intertek Webinar

24th September 2013, Basel

Extractables and Leachables Testing: An Introduction and

Strategies in Current Practice

Dr. Armin Hauk

Intertek

ECA Concept Heidelberg Extractables & Leachables Seminar

14/15 May 2014 in Copenhagen

www.intertek.com2

Extractables and Leachables (E&L) Studies

1. Introduction to Extractables and Leachables (E&L) Studies

2. The strategy for a reasonable E&L Study.

3. Illustrative examples from E&L Studies.

4. The evaluation and assessment of E&L data.

www.intertek.com3

What is an Extractables and Leachables (E&L) Study?

An E&L investigation guarantees that a pharmaceutical packaging is save and does not negatively influence the drug product. In this context also “devices” like inhalers or prefilled syringes are regarded as a primary packaging system (a “container closure system”).

• Extractables: compounds released from a container/closure system into an

extraction solution in forced extraction experiments

• Leachables: compounds that leach from a container/closure system into the

“drug formulation” under normal storage conditions (as part of

the stability study)

=> Extractables are worst case Leachables

=> Leachables are a subset of Extractables

=> Leachables are effectively applied to the patients

www.intertek.com4

Why to perform E&L Studies?

Critical examples from the past – pharmaceutical packaging area (MDI) –polyaromatic hydrocarbons (PAHs), mercapto-benzothiazole and N-nitrosamines; (prefilled syringes) tungsten case & photoinitiators.

Critical examples from the past – food packaging area – ITX and other printing compounds and N-Nitrosamines.

Scientific reasons - additives and process chemicals are small molecules, some pharmaceutical formulations are quite good solvents for such extractables

To avoid surprises - degradation products of polymeric matrix or additives. Unknown compounds from manufacturing of the packaging (“NIAS”) and contaminants. Compounds migrating from printing and adhesives into the drug products.

Regulatory Guidelines – USP, EP, FDA, EMEA

www.intertek.com5

Why to perform E&L Studies?

EU-Pharmacopoeia Chapter 3, incl. Supplement 5.1, 5.2 & 5.3

USP e.g. <381> for elastomers, <661> for polymer characterization

FDA Guidance for Industry: Container Closure Systems for Packing Human Drugs and Biologics (1999)

FDA Guidance for Industry: Metered Dose Inhaler (MDI) and Dry Powder Inhaler (DPI) Drug Products (1998)

FDA Guidance for Industry: Nasal Spray and Inhalation Solution, Suspension, and Spray Drug Products -Chemistry, Manufacturing, and Controls Documentation (2002)

Health Canada, Guidance for Industry: Pharmaceutical Quality of Inhalation and Nasal Products (2006)

EMEA CPMP/QWP/4359/03, Guideline on Plastic Immediate Packaging Materials (2005)

EMEA CPMP/QWP/2845/00, Guideline on the Pharmaceutical Quality of Inhalation and Nasal Products (2006), in collaboration with Health Canada

EMEA CPMP/QWP/158/96, Note for Guidance on Dry Powder Inhalers

ICH Q6A: Specifications: Test Procedures and Acceptance Criteria for New Drug Substances and New Drug Products: Chemical Substances (1999)

www.intertek.com6

How to design an E&L Study?

An E&L investigation is not one single study, it is at least divided in 4 subsequent major steps:

Step 1.) The critical assessment of the packaging system and the

properties of the pharmaceutical formulation plus an evaluation

of the guidelines.

Step 2.) The Extractables Study is a set of forced lab experiments to

extract as much as possible out of the packaging material (but

not to destroy the material). In this case the chemists & polymer

chemists “creativity” is necessary to design reasonable

experiments (..).

Step 3.) The data evaluation of the extractable study, including a tox.-

assessment. Selection of critical leachables.

Step 4.) The Leachables Study is finally performed as part of the stability

study for the drug product after appropriate method optimisation

& validation for the selected leachables.

Step 2b:

“Migration”

Eventually plus

extractables

experiments

with simulants

in original

packaging

system under

worst case

conditions &

long term

storage

www.intertek.com7


Step 1.) The critical assessment of the packaging system means to sample all available data regarding the whole packaging system (here, be investigative and be suspicious). To be taken into account:

1) The Polymer Matrix: PP, PE, LDPE, HDPE, COC,

PVC, PET, PU, PC, PA, Teflon, elastomers (rubber),

laminated systems etc.

2) Additives: e.g. phenolic antioxidants, organo-

phosphites, HALS etc.

3) Process Chemicals – dependent on the polymer and

the process

4) Printings: Solvents, binder, reactive compounds, fillers,

dyes & pigments, additives

5) Adhesives: Solvents, polymer-systems, reactive

compounds, fillers and additives.

6) Secondary Packaging Systems: Paper, cardboards,

stickers, over-wrapping foils, etc.

www.intertek.com8


Step 1.) The critical assessment of the packaging system means to sample all available data regarding the whole packaging system (here, be investigative and be suspicious). To be taken into account:

The Special Case: Elastomers, especially Rubber Materials:

A quantitative analysis of 2-Mercaptobenzothiazole, 2,2’-

Dithiobis(benzothiazole), PAHs and N-Nitrosamines

is necessary (PQRI recommendation)

The Special Case: Inner coatings of metal devices (tubes):

Quantitative analysis of BADGE and its derivatives is required

(recommended by BfArM.)

Gamma- and Steam-Sterilisation Processes:

Significant difference between polymers regarding

sterilisation treatments. Samples with and without

sterilisation shall be investigated

S

NSH

S

NS S

NS

N

CH3

CH3

N O

www.intertek.com9


Step 1.) The critical assessment of properties of the drug product means to establish a qualitative risk based approach for the possible interaction between packaging material and drug product

Potential interaction of packaging with drug formulation

Degree of concern

High Medium Low

High Inhalation (e.g. MDIs - organic solvents!), injectables

Sterile powders and inhalation powders

Medium

Ophthalmic solutions, transdermal ointments & patches, nasal aerosols and sprays

Low

Topical solutions and suspensions, oral solutions, lingual aerosols

Oral powders Oral tablets, capsules

www.intertek.com10


Step 1.) The critical assessment of properties of the drug product means to establish a qualitative risk based approach for the possible interaction between packaging material and drug product

www.intertek.com11

How to design E&L Studies?

Step 1.) The critical assessment of properties of the drug product means to establish a quantitative risk based approach for the possible interaction between packaging material and drug product

Estimation of an analytical evaluation threshold (AET) for a MDI:

• Lowest acceptable daily dose for an extractables and leachables compound –

taking also unknowns into account - is 150 ng/day (based on PQRI).

• A typical volume of a MDI is 15 mL

• The application scenario is 0.1 mL in 3 doses per day = 0.3 mL/day

• 150 ng/day / 0.3 mL/day = 500ng/mL (concentration of extractables)

• 500ng/mL x 15 mL = 7500 ng = 7.5 µg (total released amount of an extractable)

• Assuming a part has a weight of 1 g ⇒ 7.5 µg/g (ppm) of an extractables have

to be analysable with the applied method.

• Quantitation limit for Extractables has to be better than 7.5 ppm (this is a typical

value in Extractable Studies).

www.intertek.com12

How to perform E&L Studies?

Standard set for the extraction with organic solvents of different polarity: Hexane,

Dichloromethane, Iso-propanol, Ethanol etc. plus temperature stress.

Extraction performed under reflux, in Soxleths or ultrasonic baths

(analysis with GC/MS). Caution with volatile compounds!

Standard set for aqueous extraction: Different pH-buffers, variation in salinity plus

temperature stress eventually with addition of detergents. Extraction

performed under reflux or in ultrasonic baths (analysis with LC/MS plus

GC/MS of extracts). Caution with volatile compounds and degradation

reactions!

Alternative methods, e.g.: Accelerated solvent extraction (ASE), supercritical fluid

extraction (SFE) or thermodesorption devices directly coupled with

GC/MS (TDS-GC/MS)

Step 2.) The Extractable Study is a set of forced lab experiments to extract as much as possible out of the material (but not to destroy the material).

www.intertek.com13


Step 2.) The Extractable Study is a set of forced lab experiments…what means worst case?

Typical pMDI: API plus excipients, pH-buffer, Ethanol (10%), propellant (40% fluorinated HC)

Prefilled syringe: API plus excipients, pH-buffer but >90% water

Tablets: API plus some excipients, stationary phase

– no solvent and no water!

Extraction Study only with single parts possible

Extraction Study with single parts or assembled device

Extraction Study with single parts or assembled device

Extraction Study with strong organic and aqueous solvents with pH adjustment

Extraction Study with aqueous alcohol and pH adjustment

Extraction Study with organic solvents or TENAX

www.intertek.com14


Step 2: Work flow for extractables analysis (method set-up to be comprehensive)

Aqueous / organic extract

HS-GC/MS

ICP/MS

LC/MS

pH 4 Extraction GC/MS

Derivatization GC/MS

pH 12 Extraction

Derivatization

GC/MS

GC/MS

TDS-GC/MS

GC/MS

Extraction w/oDerivatisation -GC/MS

Special analytics

OrganicExtract

OrganicExtract

OrganicExtract

www.intertek.com15


Step 2: Analytical devices typically used in an Extractables study:

TDS-GC/MSD → Comprehensive screening for VOCs,

oligomers, additives, additive break-down products

and unknowns.

Headspace-GC/MSD → Qualitative & quantitative analysis of

residual solvents & volatiles in aqueous extracts.

GC/MSD → Qualitative & quantitative analysis of extracts &

derivatised extracts, specific analysis of oligomes,

additives, & additive break-down products.

LC/MS → Quantitative analysis of extracts for semi-polar

and polar compounds - screening for polar

unknowns possible, but no perfect tool for

qualitative analysis

ICP-MS → Metals from catalysts, dyes and pigments.

NMR / MS / IR → Identification & structure elucidation of

unknowns

www.intertek.com16


Typical qualitative results of an Extractables study; here polyolefin:

2. 00 4.00 6 .0 0 8. 00 10 .0 0 1 2. 00 14.00 16. 00 1 8. 00 2 0. 00 2 2. 00 2 4.00 26. 00 28 .0 0

500 000

10 000 00

15 000 00

2000 00 0

2500 00 0

3000 00 0

3500 00 0

4000 00 0

4500 00 0

5000 00 0

5500 00 0

6000 00 0

6500 00 0

7000 00 0

7500 00 0

8000 00 0

8500 00 0

9000 00 0

9500 00 0

1 e+07

Time -->

Abu ndanc e

Minor compounds are some VOCs, some typical additives, some additive degradation products

Main compounds are sets of homologue series of PE-oligomers

O

O

O

OH

O

O

4

Irganox 10107,9-Di-tert-butyl-1-oxaspiro[4.5]deca-6,9-diene-2,8-dione

www.intertek.com17


Typical qualitative results of an Extractables study; here a polysiloxane-material:

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00

500000

1000000

1500000

2000000

2500000

3000000

3500000

4000000

4500000

5000000

5500000

6000000

6500000

7000000

7500000

8000000

8500000

9000000

9500000

1e+07

1.05e+07

1.1e+07

1.15e+07

1.2e+07

1.25e+07

1.3e+07

Time-->

Abundance

Main compounds are some non-resolved sets of homologue series of PSX-oligomers

Minor compounds are some VOCs, some additive degradation products and residuals of a polymerisation starter

O

OOH

ClCl

www.intertek.com18


“Worst case” extraction; case study 1 for non-realistic conditions

• Extractables study on multi-layer container closure system

• Extraction conditions were: aqueous solutions, 95°C, 24 h

• Sum of all identified & quantified extractables by GC/MS and LC/MS was ~20 ppm

• But the TOC of the aqueous extracts was ~120 ppm

• Reason for that difference ?

www.intertek.com19


#F:

500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600m/z

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Rela

tive A

bundance

755.7

6

711.7

4

871.8

7

799.8

4

915.9

2

959.8

9

667.7

3

843.8

5827.8

4

595.6

6

1003.9

3

1075.9

7

1031.9

1

551.6

9

639.7

1

987.9

4

1119.9

9

783.8

2

1192.0

5

1163.9

9

623.7

5

1324.0

9

1236.0

2

887.8

7

1280.0

4

1395.9

5

943.8

7

1147.9

8

1440.1

0

1252.0

2

1076.9

7

507.6

0

960.9

1

1352.0

2

1484.1

5

1441.1

4

1281.0

6

1412.0

8

683.7

4

844.8

5

800.8

4

739.7

7

756.8

4

712.8

0

1513.2

21528.1

8

1512.2

4

1557.2

31572.2

81573.2

21600.2

1

579.6

8

596.7

4

640.7

7

552.7

3

479.6

0

1613.1

3

508.6

9523.6

2

641.7

4

480.6

7

RO

R'n

RO

R'n

=> Very complex mixture of homologue series of high molecular weight compounds could be seen in the baseline of the chromatogram

• LC/ MS screening

www.intertek.com20


• “Extraction kinetics” of the TOC (H2O, 95°C):

TOC-Formation

0 5 10 15 20 25 30t [h]

TO

C

www.intertek.com21


• “Extraction kinetics” of the high molecular weight compounds (H2O, 95°C)

Formation of compounds with MW 944-988

0

50000

100000

150000

200000

250000

300000

350000

0 5 10 15 20 25 30

t [h]

LC

/MS

are

a (

neg

. m

od

e)

www.intertek.com22


• “Extraction kinetics” of low molecular weight compounds (H2O, 95°C):

RT 9.9 min, m/z = 150

0

10000

20000

30000

40000

50000

60000

70000

80000

0 5 10 15 20 25 30

t [h]

GC

/MS

are

a

RT 12.8, m/z = 73

0

20000

40000

60000

80000

100000

120000

0 5 10 15 20 25 30

t [h]

GC

/MS

are

a

www.intertek.com23



• Metal parts (MDI canister or Al-tubes) are coated with epoxy-resins.

• Problem with such coatings: BADGE and its derivatives.

• Until now data from food migration studies - carried out under standard conditions 10d at 40°C with e.g. 95% ethanol or olive oil - were accepted.

• Extractables studies were also carried out with alcohols (ethanol, iso-propanol).

• Publication from team of Mrs. Lipperheide (BfArM) showed that pharmaceutical formulations are quite better solvents for BADGE and its derivatives as alcohols and olive oil.

J. B. Haverkamp, U. Lipke, Th. Zapf, R. Galensa and C. Lipperheide

(2008) Contamination of semi-solid dosage forms by leachables from

aluminium tubes. European Journal of Pharmaceutics and

Biopharmaceutics, 70, 921-928.

www.intertek.com24



RT:0.09 - 18.13SM:7G

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Time (min)

0

10

20

30

40

50

60

70

80

90

100

Rela

tive A

bundance

0

10

20

30

40

50

60

70

80

90

100

Rela

tive A

bundance 8.21

10.5110.8910.38

9.86 11.099.7211.22

11.809.07

12.22 14.8014.30 17.6412.85 17.1815.66

7.557.32

6.435.934.814.153.320.81 2.532.07

11.72

14.8917.7616.2810.82 13.4312.3110.53

9.769.200.91 8.246.766.435.404.813.623.022.13

ACN extract

Water/iso-propanol extract (50+50)

cyclo-di-BADGE

BADGE related compounds

HPLC-APCI(+)-MS (TIC, m/z 400-2000)

cyclo-tetra-BADGE, di-BADGE*BPA,

BADGE*BuEtOH, BADGE*tBuPh

www.intertek.com25


“Comprehensive overview”, relevance of back-extractions

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.5016.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

16.50 17.00 17.50 18.00 18.50 19.00 19.50 20.00 20.50 21.00 21.50 22.00 22.50

pH of aqueous extract: pH adjustment before

organic back-extraction:

3

3

10

7

7

10

basic

acidic

acidic

basic

acidic

basic

R

O

OH

www.intertek.com26


“Comprehensive overview”, relevance of derivatised extracts

12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0

ab

c

d

e

fg h

i k

m

n

12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0

ab

c

d

e

fg h

i k

m

n

Trimethylsilyl erucamide (unsat. C22 fatty acid amide)n

Octadecanoic acid 2,3-bis[(trimethylsilyl)oxy]propyl esterm

unknown (m/z = 73, 191, 223, 257, 433, 507, 536)k

Hexadecanoic acid 2,3-bis[(trimethylsilyl)oxy]propyl esteri

Trimethylsilyl linoleic acid amideh

Trimethylsilyl oleic acid amideg

Trimethylsilyl octadecanoatef

Trimethylsilyl hexadecenoic acid amidee

Trimethylsilyl trans-9-octadecenoated

Trimethylsilyl hexadecanoatec

Trimethylsilyl trans-9-hexadecenoateb

Trimethylsilyl tetradecanoatea

Trimethylsilyl erucamide (unsat. C22 fatty acid amide)n

Octadecanoic acid 2,3-bis[(trimethylsilyl)oxy]propyl esterm

unknown (m/z = 73, 191, 223, 257, 433, 507, 536)k

Hexadecanoic acid 2,3-bis[(trimethylsilyl)oxy]propyl esteri

Trimethylsilyl linoleic acid amideh

Trimethylsilyl oleic acid amideg

Trimethylsilyl octadecanoatef

Trimethylsilyl hexadecenoic acid amidee

Trimethylsilyl trans-9-octadecenoated

Trimethylsilyl hexadecanoatec

Trimethylsilyl trans-9-hexadecenoateb

Trimethylsilyl tetradecanoatea

NH

O

Si

NH

O

Si

Trimethylsilyl oleic acid amide

Trimethylsilyl linoleic acid amide

F3C

N

O SiMe3

SiMe3

BSTFA (N,O-bis(trimethylsilyl)- trifluoroacetamide)

www.intertek.com27


igloo stopper silicone tubingsilicone gaskets

rubber gaskets

syringe plungersinjection/infusion stoppers

www.intertek.com28

Example for results from an Extractables study: Rubber material

2.00 4.00 6.00 8.00 10.0012.0014.0016.0018.0020.0022.0024.0026.00

14.00 15.00 16.00 17.00 18.00 19.00 20.00 21.00 22.00 23.00 24.00

10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00

10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00

10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00

10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00

m/z = 55

m/z = 57

m/z = 97

m/z = 99

n

n

55

57

97

99

isopropyl

end group

tert-butyl

end group


www.intertek.com29

Br

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00

500000

1000000

1500000

2000000

2500000

3000000

3500000

4000000

4500000

5000000

5500000

6000000

6500000

7000000

7500000

8000000

8500000

9000000

9500000

1e+07

1.05e+07

1.1e+07

Time-->

Abundance

TIC: 11124_6720GC_1a.D\data.ms

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00

500000

1000000

1500000

2000000

2500000

3000000

3500000

4000000

4500000

5000000

5500000

6000000

6500000

7000000

7500000

8000000

8500000

9000000

9500000

1e+07

1.05e+07

1.1e+07

Time-->

Abundance

TIC: 11124_6722GS_1a.D\data.ms

Thermodesorption-GC/MS chromatograms of two bromobutyl stoppers

BHT

C21H40

Isomers of C21H39Br

C13H24

Isomers of C13H23Br

C21H40

Isomers of C21H39Br

C13H24

Isomers of C13H23Br

Hexane Methylcyclopentane Cyclohexane

Hexane Methylcyclopentane Cyclohexane



www.intertek.com30


MercaptobenzothiazoleOrganic rubber extract after back extraction


www.intertek.com31

Quantification approaches in an Extractables study:

• Individual quantification with references

• Quantification by a qualified medium response factor

• Quantification by the “bottom line” approach

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00

500000

1000000

1500000

2000000

2500000

3000000

3500000

4000000

4500000

5000000

5500000

6000000

6500000

7000000

7500000

8000000

8500000

9000000

9500000

1e+07

Time-->

Abundance


www.intertek.com32

Leachabels Check

Experiment, e.g. with

placebo, under conditions of

accelareted stabi.-test

Extraction experiment 2 – more realistic set-up, e.g. with organic/aquoeous solvents

Extraction experiment 1 – worst case extractions, e.g. with strong solvents

TDS-GC/MS screening, comprehensive overview of Extractables

Step 3.) The data evaluation of the extractable study, including a tox.-assessment. Finally selection of critical leachables.

Final list of relevant

extractables

How to interpret Extractables Results?

www.intertek.com33

Daily dose depended

assessment

With published tox-data

With estimated or derived tox-data

Final list of relevant

extractables

Step 3.) The data evaluation of the extractable study, including a tox.-assessment. Finally selection of critical leachables.


www.intertek.com34


Step 3.) The data evaluation of the extractable study, including a tox.-assessment. Selection of critical leachables.

Threshold values from Guidelines which can be applied to Extractables

• PQRI: "Justification of Thresholds for Leachables in Orally Inhaled and Nasal

Drug Products“; a Safety Concern Threshold SCT is defined for potential

carcinogenic compounds with 0.15µg/day and for non-carcinogenic 5µg/day.

• EMAE: CHPM/QWP/251344/2006; Guideline on the Limits of Genotoxic

Impurities; a Threshold of Toxicological Concern (TTC) is given with

1.5µg/day intake.

• EMEA: CHMP/SWP/QWP/4446/00corr.; Guideline on the Specification

Limits for Residues of Metal Catalysts; three classes of Me types and limit

values (as dose) are given.

• Solvents according to limits for residual solvents given in USP or Pharm-Eur.

www.intertek.com35


• 25 Tablets in one bottle

• Extractables Study: Compound (X) was found with 2 µg per bottle.

• Worst case estimation: Total transfer of compound (X) to pharmaceutical formulation ⇒ 2 µg in 25 tablets.

• Maximum 3 tablets/day ⇒ (2 µg/25) x 3 = 0.24 µg

• Result: Depending on tox-profile of compound X most probably acceptable – E&L studies not necessary

www.intertek.com36


• Infusion bag, 250 mL volume

• Results form an extractables study: Compound (X) was found with a concentration of 20 ppb in the infusion solution ⇒ 20 µg/L compound (X) in infusion solution.

• Maximum 3 bags/day ⇒ 15 µg of compound (X) is applied directly by injection.

• Result: Depending on tox-profile of compound (X) this could be an acceptable or not-acceptable dose – leachables study most probably necessary.

www.intertek.com37

Case Study: IRGANOX 1010

OH

O

O

OH

O

O

OHO

O

OH

OO

Molecular weight: 1177.7

Water solubility: <0.1 mg/l (at 20°C)


www.intertek.com38


Sources of toxicology data identified

• Ciba, 2000, IRGANOX 1010: Tetrakis-(Methylene-(3,5-di-tertbutyl-4-hydrocinnamate)Methane. CAS NO. 6683-19-8. Submitted by: Ciba Specialty Chemicals Corporation; New York to United States Environmental Protection Agency (EPA); Washington, DC.

• Drake, G.A.; London, J.E.; Smith, D.M.; Thomas, R.G. 1980. Preliminary Toxicological Study of Irganox 1010. Prepared by: University of California, Los Alamos Scientific Laboratory; Los Alamos, New Mexico for the US Department of Energy; Washington, DC. [LA-8037-MS].

• European Commission, 2000, IUCLID Data Set CAS No. 6683-19-8.


www.intertek.com39


Acute toxicity: oral LD50 = >10,250 mg/ kg body weight; inhalation LC50 = >1,951 mg/m3 (>2,070 mg/kg body weight)

Skin Irritation: Non-irritating (rabbits)

Sensitization: Not sensitizing (guinea pigs)

Eye Irritation: Mild, but transitory, eye irritant (rabbit)

Subchronic toxicity

• 13-week dietary study in beagles: NOAEL = 10,000 ppm (~250 mg/kg/day)

Carcinogenicity

• Dietary studies in rats and mice; no indication of tumorigenic potential in either species

Genotoxicity

• Not mutagenic in Ames assay

• Not clastogenic in in vivo bone marrow chromosomal aberration test in Chinese hamsters

Reproductive and Developmental toxicity

• Two generation reproductive dietary study in rats; no effects on reproductive capacity; no maternal toxicity; NOAEL 10,000 ppm – highest dose tested (~1,000 mg/kg/day)

• Teratology studies - oral gavage in rats; NOAEL for both maternal toxicity and teratogenicity = 1,000 mg/kg/day, the highest dose tested


www.intertek.com40


Risk Characterization

• In the absence of a regulatory agency derived acceptable

daily intake, the risk characterization was based on the

lowest NOAEL observed of 250 mg/kg/d in the 13 week dog

study

• Exposure to lrganox 1010 from use of an injectable was

estimated to be 0.4 µg/kg body weight/day

• Margin of Safety 625,000

• [250 mg/kg b.w./day / 0.0004 mg/kg b.w./day

Conclusion: no clinically relevant adverse effects expected


www.intertek.com41

Step 4.) The Leachable Study is finally performed as part of the stability study for the drug product after appropriate method optimisation &

validation for the selected leachables.

Extractables versus Leachables Studies

www.intertek.com42

Extractables versus Leachables Studies

Extractables Studies are “worst case” investigations, are only partially compound-specific, the polymer chemists know-how and creativity in the lab work is valuable, a high degree of experimental freedom exists. In Extractables Studiesexperimental modifications & repetitions are possible and the ways to achieve an appropriate screening of extractables are variable.Extractables Studies should be adjusted to the drug formulation and the packaging system.

Leachables Studies simulate realistic storage conditions with the drug product (e.g. as part of the stability test protocol).Leachables Studies are compound specific, the methods have to be optimised and validated. Compounds which are monitored as leachables are potential drug impurities. Leachables Studies are performed under full QC. There is no experimental freedom & only a limited number of samples are available (e.g. out of the stability test). Qualified references are necessary. Problem: Unexpected results and/or OOS-cases.Leachables Studies are specific regarding the drug formulation and the packaging system plus the packaging size.

www.intertek.com43

Leachables Study

The method have to be validated according to ICH Q2A/Q2B:

1) Specificity - Spike experiments with authentic reference materials - GC/MS, retention time (high conc.)

2) Linearity – measurement of pure compounds and matrix spiked extracts

3) Accuracy – by spiking / comparison of linearity of pure references versus matrix spiked and extracted references

4) Precision (Reproducibility) - multiple extractions spiked and non-spiked

5) Robustness – method stability over time and extract stability

6) LOQ/LOD – determination by spiking experiments (low conc.)

www.intertek.com44

Intertek Chemicals & Pharmaceuticals

Thank you for your attention !

Questions & Discussion

Dr. Armin Hauk Tel.: +41 61 686 48 56 e-mail: [email protected]

Extractables and Leachables Testing: An Introduction...

Documents

Transcript of Extractables and Leachables Testing: An Introduction...