Extractables and Leachables from Single Use Bioprocess Systems

Transcription

1 Extractables and Leachables from Single Use Bioprocess Systems 1

2 Founded in 1980 Medical Devices Food Packaging P harmaceutical Over Extractables Leachables 1,000 Projects Annually IDENTIFICATION QUANTIFICATION A Leader & in Plastics Analysis Of Employees Are Degreed 80% Chemists Investigative ANALYSIS P olyme r n & Additive ID Deformulation Off-Odor Crac 2

3 Overview E&L Background E&L Study Overview Model Device Extraction Conditions Scouting Methods Qual Methods Scouting Method Results Qualitative Results Mass Spectroscopy Software Tools Method Sensitivity 3

4 What are Leachables & Extractables? USP 1663 & 1664 Leachables are : organic and inorganic chemical entities that migrate from a packaging/delivery system, packaging component, or packaging material of construction into an associated drug product under normal conditions of storage and use or during accelerated drug product stability studies. Extractables are: Organic and inorganic chemical entities that can be released from a pharmaceutical packaging/delivery system, packaging component, or packaging material of construction under laboratory conditions.. 4

5 Introduction to Es and Ls Examples of E&Ls Small molecules present in a polymer system including: Antioxidants Surfactants Slip agents Plasticizers Acid scavengers Cross linking agents Residual monomers and oligomers Polymerization side products Process Impurities Extractable Compounds Observed Leachables Extractables may not be leachables depending on their solubility and the use conditions of the device. 5

6 The Regulatory Landscape Pharmaceutical Packaging Pharma/Bio Manufacturing USP <661> USP <1663> USP <1664> USP <661.3> BPOG BPSA 6

7 E&L Study Background Information Device Composition Use Conditions Study Design Protocol Analysis Protocol Techniques Sample Selection Extraction Conditions Steps in an Extractables and Leachables Study Sample and Control Preparation Chemical Analysis Extraction Identification Quantitation Method Development Toxicology Toxicological Evaluation 7

8 Bioprocess System Three Component System Single Use Bioprocess Bag (LDPE and EVA) Nylon Filter with Polypropylene Filter Housing Thermoplastic Elastomer Tubing 8

9 E&L Study Background Information Device Composition Use Conditions Study Design Protocol Analysis Protocol Techniques Sample Selection Extraction Conditions Steps in an Extractables and Leachables Study Sample and Control Preparation Chemical Analysis Extraction Identification Quantitation Method Development Toxicology Toxicological Evaluation 9

10 Single Use Bioprocess System Extraction Conditions Extractables Cut and Cover 50 C for 72hr at 50 RPM Leachables Continuous Recirculation at 37 C for 72hr 10

11 Extraction Solvents and Samples Saline Ethanol Ethanol\Water 50/50 Water Bioprocess Bag X X X X Pharma Tubing X X X X Disk Filter X X X X Complete System X Saline = 137 mm NaCl, 2.7 mm KCl and 10 mm phosphate buffer solution (ph 7.4) 11

12 Scouting Methods Bulk Identification FTIR Grav Non-Volatile Residue Scouting Methods Conjugation Double Bonds UV-Vis TOC Organic Carbon 12

13 Instrumental Techniques Technique Analyte, Boiling Point DHS-MS QTOF-GCMS QTOF-LCMS Very Volatile Compounds (VVOC) <100 C Volatile Compounds (VOC) C Semi Volatiles (SVOC) C Non-Volatile and Ionizable ICP-MS Elemental Analysis 13

14 Analytical Sensitivity Analytical Evaluation Threshold Must Identify AET 50 ng/ml Must Identify AET 25 ng/ml 14

15 Analytical Evaluation Threshold (AET) The threshold at or above which a particular leachable and/or extractable should be identified Drug Products vs Medical Devices Relative value based on: The Safety Concern Threshold (SCT) The drug product configuration Analytical techniques/methods used The Effects of Standard Selection 15

16 AET Value Analytical Evaluation Threshold Erucamide 25 ng/ml Irganox ng/ml 16

17 Compound Standard Erucamide Irganox 1010 Molecular Formula RRF Percent of True Value RRF Percent of True Value Erucamide C22H43NO % % Oleamide C18H35NO % % Octadecanamide C18H37NO % % Dioctyl phthalate C24H38O % % mono-butyl phthalate C12H14O % % Benzyl butyl phthalate C19H20O % % Didecyl phthalate C28H46O % % Irganox 1010 C73H108O % % Irganox 1135 C25H42O % % Irganox 259 C40H62O % % Irganox 3114 C48H69N3O % % Irganox 1141 C24H42O % % Caprolactam C6H11NO % % Triallyl-triazine-trione C12H15N3O % % 17

18 Detector MS UV-VIS CAD Compound Molecular % of True % of True % of True RRF RRF RRF Formula Value Value Value Fatty Amides Erucamide C22H43NO % % Oleamide C18H35NO % % Octadecanamide C18H37NO % % Phthalates Dioctyl phthalate C24H38O % % % mono-butyl phthalate C12H14O % % % Benzyl butyl phthalate C19H20O % % % Didecyl phthalate C28H46O % % % Antioxidants Irganox 1010 C73H108O % % % Irganox 1135 C25H42O % % % Irganox 259 C40H62O % % % Irganox 3114 C48H69N3O % % % Irganox 1141 C24H42O % % % 18

19 Relative Quantitation Results MS UV-VIS CAD Compound Class Mean RRF STD. Dev. % RSD Mean RRF STD. Dev. % RSD Mean RRF STD. Dev. % RSD Fatty Amides % % Phthalates % % % Antioxidants % % % All Compounds % %* % Optimizing Standard and Detector Type Significantly Reduces Response Factor Variation * Value excludes Fatty Amides which have no chromophore. 19

20 E&L Study Background Information Device Composition Use Conditions Study Design Protocol Analysis Protocol Techniques Sample Selection Extraction Conditions Steps in an Extractables and Leachables Study Sample and Control Preparation Chemical Analysis Extraction Identification Quantitation Method Development Toxicology Toxicological Evaluation 20

21 Gravimetric Analysis & TOC The Foundational Methods for Extractables Quantitation Sample Bag Tubing Filter Solvent Corrected Residue TOC (mg/device) (mg/device) Water /50 Water/Ethanol Ethanol Saline Water /50 Water/Ethanol Ethanol Saline Water /50 Water/Ethanol Ethanol Saline

22 FTIR Scouting Methods Functional Group Identification Blue = Ethanol Tubing Extract Red = Mineral Oil 22

23 UV-VIS Scouting Methods Chromophore Containing Species UV Absorbance 23

24 Background Information Study Design Protocol Steps in an Extractables and Leachables Study Device Composition Analysis Protocol Techniques Chemical Analysis Quantitation Toxicology Sample Selection Sample and Control Preparation Extraction Method Development Toxicological Evaluation Use Conditions Identification Extraction Conditions 24

25 Extraction Scouting Qualitative Analysis QTOF-GCMS ICP-MS QTOF-LCMS DHGCMS Identification 25

26 Dynamic Headspace Mass Spectrometry Very Volatile Compounds (VVOC) <100 C 26

27 ICP-MS Saline Water Elemental Analysis As Cd Hg Pb Co Ni Cr Mo Sb Ba Class (ICH Guidelines) A 2A Filters <DL <DL <DL <DL Tube <DL <DL 1.34 <DL Bag <DL <DL <DL Flow Through <DL <DL Filters <DL <DL <DL <DL <DL Bag <DL <DL <DL <DL <DL <DL Tube <DL <DL <DL Results in ng per Device 27

28 ICP-MS Saline Water Saline Extracts Na 350,000 ng/ml As Cd Hg Pb Co Ni Cr Mo Sb Ba Class (ICH Guidelines) A 2A Filters <DL <DL <DL <DL Tube <DL <DL 1.34 <DL Bag <DL <DL <DL Flow Through <DL <DL Filters <DL <DL <DL <DL <DL Bag <DL <DL <DL <DL <DL <DL Tube <DL <DL <DL Results in ng per Device 28

29 QTOF-GCMS: Bioprocess Bag Extractables Ethanol 50% Ethanol Water Agilent 7200 QTOF GCMS EI; DB-5MS 29

30 QTOF-LCMS: Bioprocess Bag Extractables Ethanol 50%Ethanol Water Agilent 6545 QTOF LCMS ESI positive ionization 30

31 QTOF LCMS Ethanol Extract of Bioprocess Bag 10 7 Counts Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, Positive Ionization 31

32 QTOF LCMS Ethanol Extract of Bioprocess Bag 10 6 Counts Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI,Positive Ionization 32

33 QTOF LCMS Ethanol Extract of Bioprocess Bag 10 5 Counts Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI,Positive Ionization 33

34 Agilent Profinder Feature (Compound) Finding Using Recursive Analysis Agilent Mass Profiler Professional Filtering Statistical Analysis 34

35 MS Advanced Qualitative Workflow Find Compounds Compare ID Compounds Recursive Feature Finding Statistical Analysis Fold Change Filtering Abundance Filtering Formula Generation Databases 35

36 Advanced Qualitative Workflow Find Compounds Compare ID Compounds Recursive Feature Finding Statistical Analysis Fold Change Filtering Abundance Filtering Formula Generation Databases 36

37 QTOF LCMS Ethanol Extract of Bioprocess Bag 10 5 Counts Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI,Positive Ionization 37

38 Total Compounds Detected QTOF LCMS Tubing Filter Bag Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI + Ionization 38

39 Advanced Qualitative Workflow Find Compounds Compare ID Compounds Recursive Feature Finding Statistical Analysis Fold Change Filtering Abundance Filtering Formula Generation Databases 39

40 Bioprocess Bag Ethanol Extract Recursive Feature Finding Agilent Profinder 726 Compounds Agilent Mass Profiler Professional 575 Extractable Compounds Significance and Fold Change Filtering 288 Compounds from Control Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, + Ionization 40

41 Advanced Qualitative Workflow Find Compounds Compare ID Compounds Recursive Feature Finding Statistical Analysis Fold Change Filtering Abundance Filtering Formula Generation Databases 41

42 With this many Compounds which areimportant? 42

43 Analytical Evaluation Threshold SCT = 0.15 µg/day Safety Concern Threshold Each system provides 500 doses Each patient takes 2 doses/day 0.15 µgg dddddd 1 dddddd 2 dddddddddd 500 dddddddddd SSSSSSSSSSSS AET =37.5 µg/device Estimated Analytical Evaluation Threshold 43

44 44 Estimated AET =37.5 µg/device

45 Adjusting AET AET =37.5 µg/device Estimated Analytical Evaluation Threshold Mean Relative Response Factor = 0.78 Standard Deviation = %RSD = 83% 37.5 µgg DDDDDDDDDDDD µgg DDDDDDDDDDDD 0.83 AET = 6.4 µg/device Final Analytical Evaluation Threshold 45

46 Estimated AET =37.5 µg/device Final AET =6.4 µg/device 46

47 47 Abundance Filtering at AET

48 Advanced Qualitative Workflow Find Compounds Compare ID Compounds Recursive Feature Finding Statistical Analysis Fold Change Filtering Abundance Filtering Formula Generation Databases 48

49 Identification Jordi Proprietary Additives and Polymer Database Agilent E&L Database Molecular Formula Generation Confirmation High Resolution MSMS Fragmentation Spectra Analysis of Reference Standards 49

50 Identification Erucylamide C 22 H 43 NO Trioctyl Trimellitate C 33 H 54 O 6 Oxidized Irgafos 168 C 42 H 63 O 4 P Caprolactam C 6 H 11 NO Oxidized Irgafos 168 Degradants Var. Hexanamide C 6 H 13 NO Caprolactam Dimer C 12 H 22 N 2 O 2 Tetradecamethylcyclohexasiloxane C 14 H 42 O 7 Si 7 Docosanamide C 22 H 45 NO Oleylmorpholine C 22 H 43 NO Eicosenamide C 20 H 39 NO Pentamethylpiperidinol C 10 H 21 NO Pentanamide C 5 H 11 NO Azacyclododecan-2-one C 11 H 19 NO - 50% Ethanol - Water - Saline 50

51 Management of large datasets through Statistical Filtering Background and Trace Compounds Removed Identifications made using Database Searching & Molecular Formula Generation 51

52 Tubing Extractables by Solvent 50% Ethanol Ethanol Water Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, Positive Ionization 52

53 Tubing Extractables by Solvent 50% Ethanol Ethanol Water Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, Positive Ionization 53

54 Tubing Extractables vs. Saline Extractables Saline Extract Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, Positive Ionization 54

55 Tubing Extractables vs. Saline Extractables Saline Extract Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, Positive Ionization 55

56 Individual Parts vs Flow Extraction Individual Parts Flow Through Extraction Agilent 1290 UHPLC Agilent 6545 QTOF LCMS Dual ESI, Positive Ionization 56

57 Bioprocess System Extract Preparation Scouting Analyses by FTIR, TOC, Gravimetric and UV-VIS Qualitative Screening (DHS, ICP-MS, QTOF-MS) Unknown Identification Feature Finding, Statistical Analysis and Abundance Filtering using Agilent Profinder and MPP Unknown Identification by Database and MFG 57