Best Practices for OINDP Pharmaceutical Development Programs Leachables and Extractables V. The Controlled Extraction Study PQRI Leachables & Extractables Working Group

PQRI Training Course September 20-21, 2006 Washington, DC

Definition

► A Controlled Extraction Study is a laboratory investigation into the qualitative and quantitative nature of extractable profiles from critical components of an OINDP container/closure system

PQRI Safety Thresholds and Best Practices for Extractables and Leachables in OINDP November 2005

Top Ten Reasons Why Controlled Extraction Studies are Needed

► ► ► ► ► To make an informed selection of materials.

To meet regulatory expectations.

► ► To control leachables.

To control materials from lot to lot.

► To correlate extractables data to leachables.

► ► To evaluate the safety of the materials. To predict worst case of end of shelf life leachables.

To qualify packaging materials.

To obtain a comprehensive extractables profile. Because USP testing does not provide applicable data.

Course Objectives

► Purpose of a Controlled Extraction Study ► PQRI Best Practice Recommendations  Controlled Extraction Study Example Data ► Qualitative and Quantitative Profiles ► Method Optimization ► Conclusion

The Purpose of a Controlled Extraction Study is to systematically and rationally identify and quantify potential leachables, to the extent practical, and within certain defined analytical threshold parameters.

Utility of Extractable Information

► Obtain Data for Risk Assessment   Provide Information to Toxicologists for Preliminary Risk Assessment Apply Threshold Principles ► Provide Basis for Leachable Methods ► Correlate Extractable Data to Leachables Data ► Develop Routine Extractable Tests ► Establish Control Criteria

Study Strategy

► ► ► When to Begin  Early in Development Phase ► Establish Team and Obtain Extractable Information Where to Begin  Select Critical Components ► Knowledge of Materials   Extraction Solvents/Techniques Analytical Methods Where to End  Application of the AET   Identification Categories Data Evaluation and Reporting  Control of Leachables

Critical Components

► MDI, DPI, Nasal Inhalation Solutions and Sprays      Patient Contact Product Contact Device Performance Secondary Packaging Ancillary Components

Typical Materials

► OINDP Components   Valves (Gaskets/O-rings) Mouthpiece  Canister   Secondary Packaging Pump Components   Actuator Containers   Blisters Labels/Adhesives/Inks ► Extractables (0.01-1000ug)           Solvents Monomers/Dimers/Trimers Curatives Photo Initiators Plasticizers Lubricants Processing Aids Antioxidants Cleaning Residues Reaction/Degradation and Breakdown Products

Knowledge of Materials/Processes

► Materials of Composition   Base Material Additives and Processing Aids ► Polymerization Process ► Fabrication process ► Cleaning and Pretreatment ► Component Storage and Shipping

Extractable Profiles

► Qualitative  Comprehensive ► Quantitative  Worst Case Leachables ► Component Control  Acceptance Criteria

Challenges/Choices

• What Components? • How Many Components ? • What Volume of Solvent? • What Reference Material Should be

Selected?

• What Solvents? • What Extraction Techniques? • What Analysis Conditions?

Considerations

► Extraction should be vigorous, but not so aggressive as to alter the qualitative and/or quantitative nature of the extractable profile ► Must be technically justified and optimized to produce extractables profiles at least equivalent to leachable profiles obtained under worst case conditions Jenke, DR. PDA J Pharm Sci Technology, 2003

PQRI Best Practices Recommendations

Ten Principle Objectives

#1 Controlled Extraction Studies should employ vigorous extraction with multiple solvents of varying polarity .

► ► ► ► ► ► ► Range of Polarities Range of Boiling Points One of Similar Extracting Properties to Drug Product Vehicle Relatively Non-reactive High Purity Easily and Safely Handled Readily Available

Solubility

mAU DAD1 A, Sig=200,4 Ref=550,100 (I:\HPCHEM\1\DATA\022569\022569\JAN31012.D) 300 250 200 150 100 50 0 0 2 4 6 Hexane (Reflux) 8 10 12 14 16 18 min mA U DAD1 A, Sig=200,4 Ref=550,100 (I:\HPCHEM\1\DATA\022569\022569\JAN31008.D) 17 5 15 0 12 5 10 0 75 2-propanol (Reflux) 50 25 0 0 2 4 6 8 10 12 14 16 18 mi

Thermolysis

A b u n d a n c e 1 . 2 e + 0 7 1 . 1 5 e + 0 7 1 . 1 e + 0 7 1 . 0 5 e + 0 7 1 e + 0 7 9 5 0 0 0 0 0 9 0 0 0 0 0 0 8 5 0 0 0 0 0 8 0 0 0 0 0 0 7 5 0 0 0 0 0 7 0 0 0 0 0 0 6 5 0 0 0 0 0 6 0 0 0 0 0 0 5 5 0 0 0 0 0 5 0 0 0 0 0 0 4 5 0 0 0 0 0 4 0 0 0 0 0 0 3 5 0 0 0 0 0 3 0 0 0 0 0 0 2 5 0 0 0 0 0 2 0 0 0 0 0 0 1 5 0 0 0 0 0 1 0 0 0 0 0 0 5 0 0 0 0 0 0 T i m e - - > 5 . 0 0 A b u n d a n c e 1 0 . 0 0 T I C : 0 2 2 1 0 3 0 2 . D 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 1 . 2 e + 0 7 1 . 1 5 e + 0 7 1 . 1 e + 0 7 1 . 0 5 e + 0 7 1 e + 0 7 9 5 0 0 0 0 0 9 0 0 0 0 0 0 8 5 0 0 0 0 0 8 0 0 0 0 0 0 7 5 0 0 0 0 0 7 0 0 0 0 0 0 6 5 0 0 0 0 0 6 0 0 0 0 0 0 5 5 0 0 0 0 0 5 0 0 0 0 0 0 4 5 0 0 0 0 0 4 0 0 0 0 0 0 3 5 0 0 0 0 0 3 0 0 0 0 0 0 2 5 0 0 0 0 0 2 0 0 0 0 0 0 1 5 0 0 0 0 0 1 0 0 0 0 0 0 5 0 0 0 0 0 3 0 . 0 0 3 5 . 0 0 0 T i m e - - > 5 . 0 0 Methylene Chloride T I C : 0 2 2 7 0 3 0 3 . D A b u n d a n c e 1 0 . 0 0 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 3 0 . 0 0 3 5 . 0 0 T i m e - - > 2-propanol 6 5 0 0 0 0 0 6 0 0 0 0 0 0 5 5 0 0 0 0 0 5 0 0 0 0 0 0 4 5 0 0 0 0 0 4 0 0 0 0 0 0 3 5 0 0 0 0 0 3 0 0 0 0 0 0 2 5 0 0 0 0 0 2 0 0 0 0 0 0 1 5 0 0 0 0 0 1 0 0 0 0 0 0 5 0 0 0 0 0 1 . 1 e + 0 7 1 . 0 5 e + 0 7 1 e + 0 7 9 5 0 0 0 0 0 9 0 0 0 0 0 0 8 5 0 0 0 0 0 8 0 0 0 0 0 0 7 5 0 0 0 0 0 7 0 0 0 0 0 0 5 . 0 0 T I C : 0 2 2 5 0 3 0 3 . D 1 0 . 0 0 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 3 0 . 0 0 3 5 . 0 0 Hexane

Extractable Yield

Abundance 2000000 1800000 1600000 1400000 1200000 1000000 800000 600000 400000 200000 Time--> 0 5.00 TIC: 02280334.D Abundance 10.00 15.00 20.00 25.00 30.00 1300000 1200000 1100000 1000000 900000 800000 700000 600000 500000 400000 300000 200000 100000 35.00 Time--> 0 5.00 Methylene Chloride TIC: 02280338.D 10.00 15.00 20.00 25.00 30.00 2-propanol Abundance 1800000 1600000 1400000 1200000 1000000 800000 600000 400000 200000 0 5.00 10.00 15.00 Hexane TIC: 02280342.D 20.00 25.00 30.00 35.00

#2 Controlled Extraction Studies should incorporate multiple extraction techniques .

Sulfur Cured Elastomer

A b u n d a n c e A b u n d a n c e 4 5 0 0 0 0 0 4 0 0 0 0 0 0 3 5 0 0 0 0 0 3 0 0 0 0 0 0 2 5 0 0 0 0 0 2 0 0 0 0 0 0 1 5 0 0 0 0 0 1 0 0 0 0 0 0 T i m e - - > 5 0 0 0 0 0 0 5 . 0 0 1 0 . 0 0 T I C : 0 2 0 3 0 3 0 4 . D 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 1 . 3 e + 0 7 1 . 2 5 e + 0 7 1 . 2 e + 0 7 1 . 1 5 e + 0 7 1 . 1 e + 0 7 1 . 0 5 e + 0 7 1 e + 0 7 9 5 0 0 0 0 0 9 0 0 0 0 0 0 8 5 0 0 0 0 0 8 0 0 0 0 0 0 7 5 0 0 0 0 0 7 0 0 0 0 0 0 6 5 0 0 0 0 0 6 0 0 0 0 0 0 5 5 0 0 0 0 0 5 0 0 0 0 0 0 4 5 0 0 0 0 0 4 0 0 0 0 0 0 3 5 0 0 0 0 0 3 0 0 0 0 0 0 2 5 0 0 0 0 0 2 0 0 0 0 0 0 1 5 0 0 0 0 0 1 0 0 0 0 0 0 5 0 0 0 0 0 3 0 . 0 0 T i m e - - > 5 . 0 0 Sonication T I C : 0 3 1 4 0 3 0 3 . D 1 0 . 0 0 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 Soxhlet Abundance 6000000 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 3 0 . 0 0 3 5 . 0 0 Time--> 0 1.2e+07 1.15e+07 1.1e+07 1.05e+07 1e+07 9500000 9000000 8500000 8000000 7500000 7000000 6500000 5.00

TIC: 02270303.D

10.00

15.00

20.00

25.00

30.00

35.00

Reflux

Peroxide- Cured Elastomer

Abundance TIC: 02280309.D Abundance TIC: 02280324.D 1000000 900000 800000 700000 600000 500000 400000 300000 200000 100000 Time--> 0 5.00 10.00 Sonicatrion 15.00 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 20.00 25.00 30.00 35.00 Time--> 0 5.00 10.00 15.00 20.00 25.00 30.00 35.00 Reflux

Polypropylene

mAU 175 DAD1 A, Sig=200,4 Ref=550,100 (J:\HPCHEM\1\DATA\022569\FEB14011.D) 25 0 75 50 150 125 100 2-propanol Blank (Sonication) 125 100 25 0 75 50 mAU 175 0 2 4 6 8 DAD1 A, Sig=200,4 Ref=550,100 (J:\HPCHEM\1\DATA\022569\FEB14012.D) 10 12 022569-01 2-propanol Extract (Sonication) 150 0 2 4 6 8 10 12 14 14 16 18 min 16 18 min mAU DAD1 A, Sig=200,4 Ref=550,100 (I:\HPCHEM\1\DATA\022569\022569\JAN31008.D) 175 150 125 100 75 2-propanol (Reflux) 50 25 0 0 2 4 6 8 10 12 14 16 18 min

#3 Controlled Extraction Studies should include careful sample preparation based on knowledge of analytical techniques to be used .

► Preparation of Extracts  Sampling, Sample:Surface Ratio, Solvents, Conditions ► Test Sample Preparations   Instrumental Techniques Concentration/Dilution

A b u n d a n c e T i m e - - > 1 . 2 e + 0 7 1 . 1 5 e + 0 7 1 . 1 e + 0 7 1 . 0 5 e + 0 7 1 e + 0 7 9 5 0 0 0 0 0 9 0 0 0 0 0 0 8 5 0 0 0 0 0 8 0 0 0 0 0 0 7 5 0 0 0 0 0 7 0 0 0 0 0 0 6 5 0 0 0 0 0 6 0 0 0 0 0 0 5 5 0 0 0 0 0 5 0 0 0 0 0 0 4 5 0 0 0 0 0 4 0 0 0 0 0 0 3 5 0 0 0 0 0 3 0 0 0 0 0 0 2 5 0 0 0 0 0 2 0 0 0 0 0 0 1 5 0 0 0 0 0 1 0 0 0 0 0 0 5 0 0 0 0 0 5 . 0 0 1 0 . 0 0

Sample Introduction

Abundance TIC: 02270303.D

T I C : 0 2 2 7 0 3 0 2 . D 1.2e+07 1.15e+07 1.1e+07 1.05e+07 1e+07 9500000 9000000 8500000 8000000 7500000 7000000 6500000 6000000 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 0 5.00

10.00

15.00

20.00

1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 3 0 . 0 0 3 5 . 0 0 Time--> 25.00

30.00

35.00

Reconstituted in Methylene Chloride 2- Propanol Extract

Artifacts

► 2-(chloromethylthio)benzothiazole N S SH + CH 2 Cl 2 Heat N S S Cl + HCl

Solvent Compatibility

DAD1 A, Sig=200,4 Ref=550,100 (G:\HPCHEM\1\DATA\022569\JAN31009.D) mAU 175 150 125 100 75 50 25 0 Methylene Chloride Blank (Reflux) mAU 175 150 125 0 2 4 6 8 DAD1 A, Sig=200,4 Ref=550,100 (G:\HPCHEM\1\DATA\022569\JAN31010.D) 10 12 022569-01 Methylene Chloride (Reflux) 100 75 50 25 0 0 2 4 6 8 10 12 14 14 16 16 18 min 18 min

Solvent Compatibility

#4 Controlled Extraction Studies should employ multiple analytical techniques

► Gas Chromatography/Mass Spectrometry (GC/MS) ► Liquid Chromatography/Mass Spectrometry (LC/MS) ► Liquid Chromatography/Diode Array Detection (LC/DAD) ► Gas Chromatography/Flame Ionization Detection (GC/FID) ► Liquid Chromatography/Ultraviolet Detection (LC/UV) ► Fourier Transform Infrared Spectroscopy (FTIR) ► Inductively Coupled Plasma/Mass Spectroscopy (ICP/MS) ► Inductively Coupled Plasma/Optical Emission Spectroscopy (ICP/OES) ► Scanning Electron Microscopy. Energy Dispersive X-Ray (SEM/EDX)

Qualitative System Suitability Compound Suggested Technique Recommended Target Concentration (ug/ml) 2-Mercaptobenzothiozole GC or LC 50 Tetramethylthiuramdisulfide Butylatedhydroxytoluene GC or LC/UV GC or LC 50 50 Irganox 1010 Diphenyl Amine Bis-(2-ethylhexyl) phthalate Bis (dodecyl) phthalate Stearic Acid 2-ethylhexanol Pyrene LC LC GC or LC GC or LC GC or LC/MS GC GC or LC/UV 50 50 50 50 100 50 1

Complimentary Techniques

A b u n d a n c e T I C : J A N 3 1 1 0 8 . D 1 1 0 0 0 0 1 0 0 0 0 0 9 0 0 0 0 8 0 0 0 0 7 0 0 0 0 6 0 0 0 0 5 0 0 0 0 4 0 0 0 0 3 0 0 0 0 2 0 0 0 0 1 0 0 0 0 0 T i m e - - > 5 . 0 0 1 1 0 . 0 0 2 1 5 . 0 0 2 0 . 0 0 3 2 - p r o p a n o l R e f l u x 2 5 . 0 0 3 0 . 0 0 3 5 . 0 0 GC/MS of polypropylene 2-propanol extract: 1 = 2,6-di-methyl benzaldehyde; 2 = 2,4-di-

tert

-butylphenol; 3 = glycerol monostearate.

Complimentary Techniques

5.3

8.6

10.6

15.6

16.0

Bis(dimethylbenzylidene) sorbitol isomer Unknown Di-tert-butylphenol Tetradecanoic acid Hexadecanoic acid 18.4

19.0

19.4

20.3

21.0

Glycerol monopalmitate / Glycerol monostearate Irganox 1010 fragment Irganox 1010 related Octadecanoic acid Irganox 1010

Compound Specific Detection

Reflux PP Dis c/CH2Cl2

03270311 100 280 nm ANALOG 5.22e5

% 0 03270311 100 Scan AP- 1175 2.93e6

% 0 5.00

10.00

15.00

20.00

25.00

30.00

35.00

40.00

Time HPLC-UV chromatogram and m/z 1175 extracted ion current profile

#5 Controlled Extraction Studies should

include a defined and systematic process for identification of individual extractables

Abundance 1.35e+07 1.3e+07 1.25e+07 1.2e+07 1.15e+07 1.1e+07 1.05e+07 1e+07 9500000 9000000 8500000 8000000 7500000 7000000 6500000 6000000 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 0 TIC: 02050302.D

31 5.00

10.00

15.00

30 35 33 20.00

36 25.00

41 53 45 49 51 55 30.00

35.00

Time-->

Identification Categories

► Confirmed:   

Mass spectrometric fragmentation behavior Confirmation of molecular weight or confirmation of elemental composition Mass spectrum matches automated library or mass spectrum and chromatographic retention index match authentic specimen

► Confident: 

Sufficient data to preclude all but the most closely related structures have been obtained

► Tentative: 

Data have been obtained that are consistent with a class of molecule only

Confirmed

Peak

21.47 min

2,2’-methylene-bis-(-6 tert -butyl)-4-ethylphenol

Confirmation of Molecular Weight; Fragmentation Behavior; Mass Spectral Library Match; RT Match to Authentic Standard Abundance 1.35e+07 1.3e+07 1.25e+07 1.2e+07 1.15e+07 1.1e+07 1.05e+07 1e+07 9500000 9000000 8500000 8000000 7500000 7000000 6500000 6000000 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 0 30 Time--> 5.00

10.00

15.00

TIC: 02050302.D

31 35 33 20.00

36 25.00

41 53 45 49 51 55 30.00

35.00

Confirmed

Peak

21.47 min

2,2’-methylene-bis-(-6 tert -butyl)-4-ethylphenol

T071703005 1173 (21.561) Cm (1171:1177-(1164:1168+1201:1214)) 100 88 Scan CI+ 4.35e3

100 [M+NH 4 ] + 74 CI mass spectrum (ammonia) % 130 386 86 61 70 73 84 98 101 114 128 124 135 152 0 F07013003 1111 (21.557) Cm (1109:1112-1116:1122) 161 163 175 177 178 191 192 193 100 191 208 209 218 229 237 252 257 272 274 276 293 312 313 314 330 331 352 353 367 368 370 377 387 388 397 409 420 425 MS2 EI+ 4.86e4

163 175 178 EI mass spectrum M +. ???? % 135 0 57 65 69 77 79 85 91 119 103 105 117 133 121 60 80 100 120 141 147 155 140 160 169 179 180 180 192 193 202 215 200 220 227 237 240 255 256 257 267 260 281 295 297 280 300 312 311 313 321 320 368 339 340 353 354 360 369 370 380 400 420 m/z

Confident

Peak Benzothiazole

8.15 min

Mass Spectrometric Fragmentation Behavior; Mass Spectrum Matches Automated Library Search Abundance 1.2e+07 1.15e+07 1.1e+07 1.05e+07 1e+07 9500000 9000000 8500000 8000000 7500000 7000000 6500000 6000000 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 0 Time--> 5.00

10.00

TIC: 02270303.D

15.00

20.00

25.00

30.00

35.00

Confident

Peak Benzothiazole Abundance

8.15 min

Average of 8.145 to 8.166 min.: 02050302.D (-) 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 30 m/ z--> Abundance 40 50 50 58 63 60 69 82 70 74 80 91 90 100 108 110 120 135 130 140 #23599: Benzothiazole (CAS) $$ Vangard BT $$ 2-BENZOTHIAZO...

135 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 30 39 40 45 50 50 58 63 60 69 82 70 74 80 91 90 100 110 120 130 140 m/ z-->

Tentative Peak 15.05 min

Coumarone-indene resin related Confirmation of Elemental Composition; Mass Spectrometric Fragmentation Behavior Abundance 1.2e+07 1.15e+07 1.1e+07 1.05e+07 1e+07 9500000 9000000 8500000 8000000 7500000 7000000 6500000 6000000 5500000 5000000 4500000 4000000 3500000 3000000 2500000 2000000 1500000 1000000 500000 0 Time--> 5.00

10.00

TIC: 02270303.D

15.00

20.00

25.00

30.00

35.00

Tentative Peak 15.05 min

Coumarone-indene resin related A b u n d a n c e A v e r a g e o f 1 5 . 0 3 6 t o 1 5 . 0 6 8 m i n . : 0 2 0 5 0 3 0 2 . D 1 1 9 + ( - ) 3 4 0 0 0 3 2 0 0 0 3 0 0 0 0 2 8 0 0 0 2 6 0 0 0 2 4 0 0 0 2 2 0 0 0 2 0 0 0 0 1 8 0 0 0 1 6 0 0 0 1 4 0 0 0 1 2 0 0 0 1 0 0 0 0 8 0 0 0 6 0 0 0 4 0 0 0 2 0 0 0 0 + 9 1 + + 5 1 5 0 6 0 6 5 7 7 7 0 8 0 9 0 1 0 3 1 2 8 1 4 5 1 0 0 1 1 0 1 2 0 1 3 0 1 4 0 1 5 0 2 3 6 1 6 5 1 6 0 1 7 0 1 8 0 1 8 9 1 8 0 1 9 0 2 0 2 2 0 0 2 1 0 2 2 1 2 2 0 2 3 0 2 4 0 m / z - - > +

# 6 Controlled Extraction Study “definitive” extraction methods should be optimized.

► Asymptotic Levels ► Represent at Least Worst Case Leachables   Qualitative Quantitative ► Verification of Quantitative Results ► Basis for Development and Validation of Routine Extractable Control Methods

Sulfur Cured Rubber

Analysis: GC/MS Extraction: Soxhlet Optimization: 7g/200ml Methylene Chloride 10:1 Dilution Internal Standard 16 Hours Extraction 1 0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0 0 2 4 Phenolic Docosane Hexacosane Coumarone indene 6 8 10

Time, hours

12 14 16 18

Extraction: Analysis: Results:

Polypropylene

Reflux HPLC/UV Optimization: Solvent to Sample Ratio Analyte Solubility/Standardization Asymptotic Extraction Chromatography Conditions 1g:25ml (50/50 THF/IPA) 3 Hr Extraction

Polypropylene

Polypropylene

0 2 4 T r a T r a T r a 6 8 2 3 4 m r A m r B m r C 5 6 7

Example Chromatography Conditions

► Qualitative ► Quantitative LC/MS

Column: C18, 4.6mmx25cm, 5u Injection Vol: 10ul (1/mL/min) Mobile Phase: 75:25 (ACN:H20) 50:50 (ACN:THF) Gradient: 30 min. gradient at 100% A to 100% B hold 12 min Ionization: APCI Scan: m/z 50-1350 ,5 sec/scan

HPLC/DAD

Column: C18, 4.6mmx25cm, 5u @ 60C Injection Vol: 10ul (1/mL/min) Mobile Phase: ACN H2O Gradient: 25 min. linear gradient at 30:70 A:B to 100% A hold 5min Detector: Diode Array Detector 200nm, 220nm, bw/4nm; ref.sig 550nm, bw/100nm

Quantitation

Internal Standard

RRF = (A a C i )/(A i C a )

  A a = Area of Analyte Peak C i = Concentration of Internal  A i = Area of Internal Standard  C a = Concentration of Analyte External Standard

RF = C s /A s

  C s = Concentration of External Standard A s = Area of External Standard ► Concentration in Sample Extract

C a = A a x RF

► Concentration in Sample Extract

C a = A a x C i /A i x RRF

Total mass = C a ug/ml x Vol of extract Total mass = C a ug/ml x Vol of extract ► Extractable in Component

μg/g Extractable = Total mass (μg)/g Component

► Extractable in Component

μg/g Extractable = Total mass (μg)/g Component

System Suitability Example

Instrument Precision (%RSD) Resolution (n=6) Tailing Factor (n=6) Sensitivity (S/N of MQL) Method Repeatability (%RSD) Intermediate Precision (%RSD) Recovery ≤ 10 ≥ 2 ≤ 2 ≥ 10 ≤ 10% ≤ 10% 80-120%

Method Validation

► Acceptance Criteria   System Suitability Instrument Precision   Chromatographic Resolution Chromatographic Tailing Factor  Linearity/Range   Precision ► Method Repeatability ► Standard Sample Stability Intermediate Precision   Specificity Accuracy   Limit of Quantitation/Limit of Detection Robustness/Ruggedness

#7 During the Controlled Extraction Study process, sponsors should revisit supplier information

► Investigate   Chemical entities found in the extractable data not included in supplier information Known chemical entities not detected in the extractable

Code Letter C B A

Extractable Data

Qualitative Results Supplier Information Tetrakis (methylene(3,5-di-t-butyl-4 hydroxyhyrocinnate)) methane Phenolic Antioxidants 0.08 % Bis(2,4-di-t-butylphenyl)pentaerythritol disphosphite and di-tert butylphenol Calcium Stearate Glycerol monopalmitate/monostearate Tetradeconoic, Hexadecanoic and Octadecanoic Acids Phosphite Antioxidant 0.05% Stearate Mould Release 0.03 – 0.4% Vegetable oil 0.2 - 0.3 % N/A 3,4-dimethyldibenzylidene sorbitol Clarifier 0.2 - 0.3 % Quantitative Results 0.059% 0.045% Not Detected Not Detected N/A 0.14

Known Accelerator

Abundance 1.35e+07 1.3e+07 1.25e+07 1.15e+07 1.1e+07 1e+07 9000000 8000000 7000000 6000000 5000000 4000000 3000000 2000000 1500000 1000000 Time--> 500000 Time--> 0 0 Tetramethylthiuram Monosulfide 5.00

5.00

10.00

10.00

15.00

15.00

TIC: 02050302.D

35 TIC: 02030503.D

30

GC-MS of Methylene Chloride Soxhlet Extract

31 33 20.00

20.00

36 25.00

25.00

41 53 45 49 51 55 30.00

30.00

35.00

35.00

#8 Controlled Extraction Studies should be guided by an Analytical Evaluation Threshold (AET) that is based on an accepted safety evaluation threshold

► How low to go to identify and evaluate individual extractable

AET Extractables

► A leachable dose less than or equal to the SCT is a dose so low that there would be negligible safety concerns from toxic effects.

   Internal or External Standards can be employed to Measure the SCT level.

The sensitivity needed for the extractable and leachable methods can be postulated. Comprehensive extractable studies can be predictive of end of shelf life leachable studies

1

st

Application of the Analytical Evaluation Threshold (AET)

► Example Nasal Spray Component #1:

4 doses per day 120 doses per container 0.15g tube

► Estimate AET: (Tox Assessment Value) Convert SCT (0.15 μgTDI) to μg/container 0.15 μg/day X 120 doses/container= 4.5 μg/tube 4 doses/day 4.5μg/tube = 30μg/g 0.15g tube

How are unknowns measured?

► Estimated ► Based on Response of   Internal Standard Significant Identified Extractable Peak ► Final  Incorporate Uncertainty ► RRF Data Base ► 1%RSD or 50% of Estimated AET

Uncertainty Factor

►

Final AET

Tube (30 ug/g) 50% correction of the estimated AET = 15ug/g Estimated AET corrected for 1%RSD/35% =20ug/g

What does that mean?

Extractables ≥ The Estimated AET should be Identified to the Extent Possible

Location of the AET (Extractables)

► Extraction  2 grams (100 cm 2 ) extracted in 100mL of 2-propanol ►  AET AET response typical of UV antioxidant species ► Final AET  50% Factor DA D1 A , Sig=200,4 Ref =550,100 (061838\SEP05029.D) mA U 120 100 80 60 40 20 0 0 2 4 Estimated AET Final AET 6 8 30ug/g 15/ug/g 10 12 14 16 min

#9 Polyaromatic Hydrocarbons (PAH’s; or Polynuclear Aromatics, PNA’s), N-nitrosamines, and 2-mercaptobenzothiazole (MBT) are considered to be “special case” compounds, requiring evaluation by specific analytical techniques and technology defined threshold

Naphthalene Acenaphthylene Acenaphthene Fluorene Phenanthrene Anthracene Fluoranthene Pyrene Benzo(a)anthracene

PAHs/PNAs

Chrysene Benzo(b)fluoranthene Benzo(k)fluoranthene Benzo(e)pyrene Benzo(a)pyrene Indeno(123-cd)pyrene Dibenzo(ah)anthracene Benzo(ghi)perylene

N-nitrosamines

N-nitrosodimethylamine N-nitrosodiethylamine N-nitrosodi-n-butylamine N-nitrosomorpholine N-nitrosopiperidine N-nitrosopyrrolidine

#10 Qualitative and quantitative extractables profiles should be discussed with and reviewed by pharmaceutical development team toxicologists so that any potential safety concerns regarding individual extractables, i.e. potential leachables, are identified early in the pharmaceutical development process

Component Profiles

Qualitative ► Sampling and Preparation ► ► Multiple Solvents Sample to Surface Ratio ► ► ► Guided by the AET  Standard Reference Materials Multiple Extraction Techniques Multiple Analytical Techniques  System Suitabilities Quantitative ► Evaluate Qualitative Profiles ► ► ► Optimize Method    Extraction ► Asymptotic Levels Analysis Conditions/Calibrations Method Accuracy/Precision Uncertainty  Finalize AET Correlation   Supplier Information Leachable Studies

Controlled Extraction Study

Summary of Steps

► ► ► ► ► ►

Qualitative Profile Determine Extractable AET

 Convert SCT total daily intake to drug product relative units then associate to the mass of the component

Quantitative Profile

    Selection of Analytes Asymptotic Extractions Linear Dynamic Range Consider Special Case Compounds

Optimize Method and Determine Range and Limits

  Recovery/Repeatability Based on Techniques used in Controlled Extraction Study

Validate Methods What Next?

Routine Extractable Testing

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Test Multiple Component Lots

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Correlate to Leachables

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Establish Specification and Acceptance Criteria

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Component Control

Science Based Approach

► ► ► ► ► ► Define expectation early Understand and apply the science involved Select appropriate CCS materials and components Apply appropriate upstream controls Communicate and Collaborate starting in early stages of drug development  With-In company  With suppliers  With regulatory bodies PQRI WG proposal seems to support similar approach

Guirag Poochikian PhD. PQRI Safety Thresholds and Best Practices for Extractables and Leachables in OINDP November 2005

Summary of PQRI Recommendations

► Controlled Extraction Studies should:          employ vigorous extraction with multiple solvents incorporate multiple extraction techniques include careful sample preparation based on knowledge of analytical techniques to be used employ multiple analytical techniques define a systematic process for identification of individual extractables optimize definitive extraction techniques/methods be evaluated relative to supplier information describing formulation consider special case (PNA, MBT and nitrosoamines) separately review profiles with development team toxicologists to be alerted to safety concerns regarding individual extractables

Conclusion

► ► Extraction techniques/methods used for Controlled Extraction Studies must be technically justified and optimized to produced extractable profiles at least equivalent to leachable profiles obtained under worst case conditions of drug product use, allowing both qualitative and quantitative extractable leachable correlations Properly conducted Controlled Extraction Studies, when accomplished early in the pharmaceutical development process, permit a pharmaceutical development team to begin early evaluation of potential drug product leachables. This evaluation can alert the pharmaceutical development team to potential leachables with toxicological concern, allowing adequate time to begin appropriate safety qualification studies, or modification of CCS system.

Conclusion

► The Best Practices recommendations for Controlled Extraction studies are not meant to be prescriptive or to exclude other scientifically valid approaches, the analytical techniques/methods, or control strategies ► These recommendations represent a consensus with-in the Working Group on current best practices with-in the pharmaceutical industry and are designed to reduce the level of uncertainty with-in the pharmaceutical development process for OINDP

References

Quality Systems Approach to Pharmaceutical Current Good Manufacturing Practice Regulation; Draft Guidance for Industry US Department of Health and Human Services FDA CDER/CBER/CVM/ORA, September, 2004 Metered Dose Inhaler (MDI) and Dry Powder Inhaler (DPI) Drug Products: Chemistry Manufacturing and Control Documentation; Draft Guidance for Industry; US Department of Health and Human Services Food and Drug Administration CDER; October 1998 Nasal Spray Inhalation Solution, Suspension, and Spray Drug Products: Chemistry Manufacturing and Control Documentation, Guidance for Industry; US Department of Health and Human Services FDA CDER; July 2002, Draft Safety Thresholds and Best Practices for Extractables and Leachables in Orally Inhaled and Nasal Drug Products; and Experimental Protocols for Controlled Extraction Studies; PQRI Leachables and Extractables Working Group; Submitted July, 2006

References

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PQRI Leachable Extractable Work Shop Presentations; December 2005, www.pqri.org

        Schroeder, A.; Leachables and Extractables in OINDP: An FDA Perspective Poochikian,G.; Best Practice Recommendations: Science and Process Ball, D; Derivation and Justification of Safety Thresholds McGovern,T.; Safety Recommendations: Science and Process Feinburg,T.; Controlled Extraction Studies Norwood,D.; Development and Application of the Analytical Evaluation Threshold Paskiet,D.; Leachable Studies and Routine Extraction Studies Winkle,H.N.; Direction for Leachables and Extractables

References

ASTM Designation: D 5524-94 “Standard Test Method for Determination of Phenolic Antioxidants in high Density Polyethylene Using liquid Chromatography” J.D. Vargo and K.L. Olson “Characterization of Addtives in Plastics by Liquid Chromatography-Mass Spectrometry,” J. Chrom., pp.215-224 (1986) Jenke, DR. Nomenclature Associated with Chmeical Characterization of and Compatibility Evaluations of Medical Product Delivery Systems. PDA J Pharm Sci Technology, 57 (2), pp. 97-108, 2003