Non-volatile extractable analysis of prefilled syringes for parenteral administration of drug products

Accepted Manuscript Title: Non-volatile extractable analysis of prefilled syringes for parenteral administration of drug products Authors: Noem´ı Dorival-Garc´ıa, Iben Larsson, Jonathan Bones PII: DOI: Reference:

S0731-7085(17)30493-4 http://dx.doi.org/doi:10.1016/j.jpba.2017.05.008 PBA 11262

To appear in:

Journal of Pharmaceutical and Biomedical Analysis

Received date: Revised date: Accepted date:

27-2-2017 30-4-2017 4-5-2017

Please cite this article as: Noem´ı Dorival-Garc´ıa, Iben Larsson, Jonathan Bones, Non-volatile extractable analysis of prefilled syringes for parenteral administration of drug products, Journal of Pharmaceutical and Biomedical Analysishttp://dx.doi.org/10.1016/j.jpba.2017.05.008 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Non-volatile extractable analysis of prefilled syringes for parenteral administration of drug products Noemí Dorival–García1, Iben Larsson2 and Jonathan Bones1,3*

1

Characterisation and Comparability Laboratory, NIBRT−The National Institute for Bioprocessing Research and Training, Foster Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland. 2

Amgros I/S, Dampfærgevej 22, DK – 2100, Copenhagen, Denmark.

3

School of Chemical and Bioprocess Engineering, University College Dublin, Belfield, Dublin 4, Ireland

*

Corresponding author: email [email protected], tel +35312158100, fax +35312158116

Noemí Dorival-García: email [email protected]. Iben Larsson: email [email protected]

Keywords: extractable, prefilled syringe, stopper, solvent extraction, UHPLC–MS, formulation

1

Graphical Abstract

2

Highlights



Determination of extractables profile for prefilled syringes at several conditions



Implementation of a simulated extractable study for long-term storage



Application of a simulated immersion study for stoppers according to ISO 10993-12



Use of liquid-liquid extraction as sample treatment for aqueous-based extracts



Use of UHPLC-QToF-MS as analytical technique for extractables identification

3

Abstract

The determination of extractable profiles for single-use technologies represents an important aspect of pharmaceutical production to minimize any possible compromise in drug product quality or potential risk to patients by identifying substances that may potentially leach from such devices. An approach for the extractable assessment of prefilled syringes, a promising alternative for parenteral administration of pharmaceutical products, is described herein. Four extraction solvents were selected: a mixture 2-propanol:water (1:1), was intended to represent aggressive conditions to extract a broad spectrum of extractables, including organic additives and substances which are poorly water-soluble. Extractions with buffers at three different working pH values spanning a range standardly used in pharmaceutical formulations were also evaluated to identify substances that require specific conditions for their extraction due to their individual chemical properties. Syringes from two different brands were analysed along with their corresponding plunger stoppers. Syringes were extracted at 40oC for 4 days, the plunger stoppers were extracted with 2-propanol at 70oC for 24 hours according to ISO 10993-12:2012. Extractables were identified by UHPLC-MS on a quadrupole time of flight instrument using a non-targeted discovery strategy. A total of 25 compounds were identified, mostly polymer additives and their degradation products. The presented methodology represents a reference point for further studies focused on the characterisation of extractables and leachables from prefilled syringes.

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1. Introduction The development of pre-filled syringes (PFS’s) offers health-care professionals a promising alternative for storing and administering parenteral medications. PFS’s also enable the selfadministration of preformulated high concentration biopharmaceuticals by patients in the outpatient setting, creating an improved patient experience by negating the need for medicine administration in hospitals [1]. PFS’s offer many advantages in relation to safety and efficacy due to reduction in dosing errors, reduced risks of biological contamination, enhanced convenience and ease of use, prevention of overfill and a reduction in required medical surveillance compared to the conventional use of ampoules or vials [2]. PFS’s are gaining strong acceptance with approximately 85% of health-care professionals recommending their usage over other parenteral delivery systems [3]. PFS’s constitute one of the fastest growing markets in the drug delivery sectors [1], with worldwide sales estimated to exceed sales of $6.9 billion by 2018, growing at a compounded annual growth rate of 13.8% from 2012 to 2018 [4]. As PFS’s are primary containers, their compatibility with the housed drug product and associated formulation needs to be critically addressed to ensure no adverse effects on patient safety and drug quality [5]. A risk associated with the use of polymeric PFS’s is extractables and leachables (E&Ls) and the identification of these substances is a critical step for determining toxicological and drug quality risks [6]. The BPSA defines extractables as “compounds that migrate from any product-contact material, including when exposed to an appropriate solvent under exaggerated conditions of time and temperature“ [7]. Extractables screening is essential to provide a complete view of potential substances that may ultimately become leachables under normal conditions of use [8]. Accordingly, PFS’s should be constructed of materials that do not leach harmful compounds that patients could be exposed during treatment with the drug product housed in the PFS. Although the PFS market is dominated by glass syringes, recent development of ‘glass-like’ cyclic olefin copolymer (COC) and cyclic olefin polymer (COP) offer alternatives to glass PFS and their associated challenges, these

5

materials have excellent transparency, good moisture barrier properties, improved robustness against breakability, better ergonomic properties and adequate stability [1, 3]. Leachables can interact with a formulation component to produce the agent responsible for the adverse safety effect. A widely documented case of incompatibility probably due to leachables that led to adverse clinical effects and product failure is that of EPREX®, used by chronic renal failure patients, in a PFS format [9]. Surfactant-aided leaching of a vulcanising agent from uncoated stoppers was proposed as a mechanism contributing to harmful effects of EPREX®[10]. Another example described the interaction between an IgG2 and leached acrylic acid from the adhesive that was used to attach the needle to the glass barrel of a PFS that resulted in unwanted modification of the IgG2 [11]. Other reports have highlighted issues with drug product exposure to leached tungsten and silicone oil from the PFS [4, 5, 12]. Although compendial requirements exist for testing elastomeric and plastic components in the US and European pharmacopoeias, few protocols are available for E&Ls studies. The most relevant guidelines for E&Ls studies are the one proposed by EMA on Plastic Immediate Packaging Materials; the FDA Guidance for Industry — Container Closure Systems for Packaging Human Drugs and Biologics; and the PQRI Thresholds and Best Practices for Extractables and Leachables in Parenteral and Ophthalmic Drug Products [3]. Information regarding extractables is beneficial since the analysis of leachables in the formulated drug is challenging due to low leachable concentrations (parts-per-billion range) and matrix inference from the excess of protein and excipients present. Therefore, the determination of extractables from the PFS is much more informative to determine which extractables may ultimately become leachables [6]. Although some reports have been published [13-15], a comprehensive evaluation is still necessary to perform toxicological assessments, that may identify unsuitable syringes and eliminate the need to conduct further resource intensive quality assurance programmes. E&Ls studies continually contribute to knowledge to ensure confident implementation of PFS’s and may create collaborative opportunities for reducing/eliminating leachables and improving PFS systems [6].

6

The aim of this current study was to apply a general and reliable method using liquid-liquid extraction followed by UHPLC-MS for the determination of the non-volatile extractables profile for assembled PFS’s from two different brands and their associated plunger stoppers. Extractables were generated by treatment of the syringes with various solvents (water and water/organic mixtures, pH, ionic strength) at elevated temperature for a specified duration. The corresponding plunger stoppers were also analysed using 2-propanol as extraction solvent according to ISO 10993-12:2012. The established method can be used in further studies about identifying E&Ls, which will provide key information enabling safety assessments that address toxicology and drug quality impact for evaluating PFS’s.

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2. Materials and Methods 2.1.

Chemicals and reagents Water used throughout was from a Sartorius Stedim Biotech Arium® 61316 system (Göttigen,

Germany). All extraction solvents, acids, bases, salts for buffers preparation and general reagents used throughout the study were ACS reagent grade or better and were from Sigma-Aldrich (Wicklow, Ireland). LC–MS Optima grade water and acetonitrile were obtained from Fisher Scientific (Dublin, Ireland). Two commercial brands of syringes were evaluated in this study. A general design for a PFS is shown in Figure 1 A. The specific materials tested included two representative polymers utilized in syringe barrels (B1 = COP and B2 = polypropylene) that were siliconized. For brand B1, two different models were analysed: B1-5 and B1-50, for 5 and 50 mL syringes, respectively. The corresponding stoppers and tip caps, made of rubber, were also analysed (S1-5 and S1-50, respectively). The 5 mL syringes for brand B2 did not include rubber stoppers, the polypropylene plungers made direct contact with the solution, but also including a rubber band to help the plunger to glide (Figure 1 B). Three different models of polypropylene tip caps were tested for this brand, consisting of the same polymeric constituents but with differences in the colorant (blue, red and grey).

2.2.

Sample preparation and extraction experiments For the assembled syringes from brand B1, four different model solvents were selected: 2-

propanol:water (1:1) and buffer solutions at pH 4, 7 and 8. For brand B2, only two solvents were tested: 2-propanol:water (1:1), and buffer solution at pH 7. McIlvaine buffer [16], a citrate/phosphate buffer was used in this study, for two reasons: (i) it contains salts that are commonly used in pharmaceutical formulations for pH regulation and (ii) it is possible to use the same buffer to set for pH in a wide range, as in this study. For all syringes extractions were performed in triplicate wherein the syringes were filled with the different extraction solvents at half of their capacity, sealed with the plunger and the tip cap and incubated in an orbital shaker at 40oC 8

during 4 days to simulate long-term storage at room temperature as previously used in other studies [13]. The shaking speed was controlled for each type of syringe to ensure that the solvent constantly contacted the inner walls of the syringe. Following completion of the incubation the syringes were subsequently allowed to cool to room temperature prior to further processing. 2 mL of 2-propanol:water extracts were transferred to 2 mL microcentrifuge tubes (Eppendorf® LoBind) and evaporated by vacuum centrifugation. 2 mL of buffer extracts were transferred to 15 mL screw-cap glass conical test tubes followed by the addition of 3 mL of dichloromethane. The mixture was shaken for 3 min and centrifuged for 10 min at 3,220 × g. The organic phase was transferred to a clean glass vial and evaporated to dryness in a water bath at 50oC. Dried residues from both procedures were reconstituted with 50 µL of a mixture of ACN:ammonium formate buffer (pH 3.0; 50 mM) prior to instrumental analysis. Blanks were also prepared using the same four extraction solvents in sealed glass containers exposed to the same extraction conditions. Extraction of the plunger stoppers with 2-propanol was performed according to the International Organization of Standards guidelines, ISO 10993-12:2012, which describes sample preparation and reference materials. The extracts were prepared by putting some stoppers (2 and 6 from S1-50 and S1-5 stoppers, respectively) in glass storage bottles with caps and adding 10 mL 2propanol and extracted at 70oC for 24 hours, to meet the extraction ratio 3 cm2/mL solvent according to this technical standard. Then extracts were filtered through a 0.45 µm nylon filter and then treated as the 2-propanol extracts from syringe barrels. Blanks were also prepared.

2.3.

UHPLC-MS analysis UHPLC–MS analysis was performed using a Waters Acquity I–Class UPLC™ (Waters,

Manchester, UK), equipped with a binary solvent manager, auto-sampler and column manager. Separation of compounds was obtained with CORTECS® UPLC® C18 column (1.6 µm; 2.1 mm × 50 mm) (Waters, UK). Eluate from the chromatographic system was sampled directly into a Waters 9

Synapt G2 HDMS QToF mass spectrometer equipped with an orthogonal Z–spray™ electrospray ionization (ESI) source. Chromatographic separations were performed using a binary gradient mobile phase consisting of ammonium formate (pH 3.0; 50 mM) (solvent A) and acetonitrile (solvent B). The column was maintained at 45oC and the injection volume was 5 µL. Gradient conditions were as follows: initial mobile phase, 85% (A) for 1 min, linearly decreased to 45% (A) within 3.0 min, to 10% within 1 min and to 0% a further minute, then held for 1 min at 100% organic mobile phase. Finally, back to 85% (A) in 0.5 min and kept for 2.5 min to equilibrate the column. Total run time was 10 min. The initial flow rate was 400 µL min-1, which was increased linearly to 500 µL min-1 between the first to fourth minute and held for 3 minutes. Then the flow was decreased to 400 µL min-1 within 0.5 min and maintained at this value until the end of the run. The mass spectrometer was operated using positive and negative ionization with data independent acquisition using 1 s sequential low and high energy scans in the range of 100−1200 Da. Nitrogen was used as nebulizer, cone and desolvation gas and argon was used as a collision gas. A collision energy ramp from 25 to 90 V was used for the high energy function. Leucine enkephalin was used as the lockmass, recorded at 10 s intervals. The accurate mass and composition of the precursor ions and fragment ions were determined using Mass Lynx v 4.1 and Progenesis QI v 2.2 (Non Linear Dynamics, Newcastle, United Kingdom).

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3. Results and Discussion As the extractables profile of PFS’s are obtained under exaggerated conditions, the selection of the extraction solvent, incubation time and temperature is crucial, to ensure that the generated extractables information identifies all possible leachables that could arise under operational conditions of use. Mixtures organic solvent:water are frequently used as simulating solvents in extractables studies as pharmaceutical formulations are aqueous-based but also contain solubilising agents for solubilisation of poorly-soluble compounds. A mixture of 2-propanol:water (1:1) was selected for this study, one of the extraction solvents recommended for controlled extraction procedures by PQRI [15]. The 1:1 ratio of organic solvent:water was selected to represent the highest proportion of solubilising agents that could be found in pharmaceutical formulations [17]. pH is also a critical factor to mimic along with the presence of salts that represent typical excipients, both of which can affect the partitioning phenomenon. Therefore, buffer solutions at pH 4, 7 and 8 were also included as extraction solvents to provide important and strategic information about potential leachables in wholly aqueous formulations. Extractable data for the non-targeted UHPLCMS analysis of the evaluated syringes are summarised in Table 1, 25 compounds were identified in total.

3.1. Extractables from plunger stoppers: Plunger stoppers showed a high number of extractables (60%) in agreement with previous studies [4, 18]. Elastomeric components represent a significant source of the identified compounds due to the excess quantities of accelerators, activators, cross-linking agents and other additives are generally applied to achieve complete vulcanization of rubber components along with antioxidants and fillers. These unreacted components remain in the rubber bulk along with their associated reaction products formed during the rubber manufacturing process and may leach into the drug product. 15 compounds were identified as extractables from the two tested plunger stoppers (S1-5 and S1-50) for brand B1. Figure 2 A shows that 33.3% of these compounds were detected only in 11

plunger stopper tests (extraction according to ISO 10993-12:2012), while the remaining 66.6% was found in the extracts from assembled syringes B1, distributed as follows: 33.3% at all tested conditions and the remaining 33.3% only in 2-propanol:water (1:1) extracts. 7 compounds (Table 1), were also found in assembled syringes B2, whose source could be the rubber band that helps the plunger to glide (Figure 1 B) and which also makes contact with the solution. The identified extractables from the plunger stoppers were found in both models (S1-5 and S1-50), suggesting their construction from the same material. These substances were identified as plasticizers, lubricants and degradation products, slip agents, residues from production, flame retardants and additives for improving impact and resistance. Three siloxanes and one silicone-based compound were also found in the plunger stoppers. These compounds come from the degradation of silicone oil that is applied to plunger and the barrel. The most common form of silicone oil used in medical applications is polydimethylsiloxane and works as a lubricant allowing the plunger to glide smoothly within the barrel to expel the formulated drug product solution. As the demand for prefilled syringes and automated injection devices increases, so does the importance of understanding silicone oil. Functionally, silicone oil is not a major concern for manual injections since a nurse or doctor is capable of applying the necessary force to push the plunger to the end point. However, springs in automated devices can only provide a fixed amount of force and any unanticipated friction may cause the plunger to stall before complete drug delivery creating the requirement for silicone oil to mitigate this risk.

3.2. Extractables from assembled syringes The extractables profile of the two different brands of syringes evaluated were determined. Brand B1 showed 18 extractables (corresponding to 72%) compared to 12 compounds from brand B2 (equivalent to 48%), Table 1. As a significant number of extractables come from rubber plunger stoppers as described above, the higher number of compounds found in brand B1 can be explained as these syringes, compared to brand B2, have both plunger stoppers and tip caps made of rubber. 12

The use of the 1:1 2-propanol water mixture and the three individual buffers produced characteristic extractables profile for each brand. Figure 2 B and C show that certain compounds were extracted only with 2-propanol:water, 28% and 8% for brands B1 and B2, respectively. The majority of compounds were extracted under all tested conditions, 67% and 50% for brands B1 and B2, respectively with very few being extracted with any of the specific buffers evaluated. For assembled syringes from brand B1, 2-propanol:water is the most suitable solvent since a higher number of compounds were extracted, Figure 3 A. However, for brand B2, most compounds were extracted with buffer at pH 7, Figure 3 B, suggesting that it is not possible to make a generalization about one only solvent system for extractables testing in PFS’s. A similar extractables profile was observed for the two syringes of different volumes from brand 1 (B1-5 and B1-50), suggesting that the two syringes are made of the same polymer and produced using similar manufacturing processes. However, a difference in the extractables identified was found between both models as shown in Figure 2 B, with 17% more compounds extracted only in the B1-5 syringe and not in the B1-50 syringe. The reason for this difference is thought to arise due to dilution issues as a result of the larger volume in B1-50. For brand 2, the same extractables profile was obtained regardless the type of tip cap, thought to be due to the minimal contact of the tip cap with the solvents during the incubation. While many of the extractable compounds identified were exclusive of each brand, barrels from both brands shared a number of common compounds as indicated in Table 1. Compounds found in the syringe barrels included mainly lubricants, degradation products of antioxidants, coatings and residues from the polymer manufacturing process. Many of the identified substances, including carboxylic acids-, amides- and esters-based additives are in agreement with previous studies [14]. Phthalates, used as plasticizers, were also identified [14]. Phthalates are known as endocrine disruptors with exposure to phthalates associated with altered hormone levels, reproductive effects (male fertility), premature puberty in pubertal girls, increased incidence of chronic disease and potentially the development of cancer [19]. 13

From the four detected degradation products from silicone oil, only one was found in barrel B1 and none in B2, suggesting that good curing of silicone, the reaction of the silicone with the syringe materials, were applied for both brands of syringes. As these compounds were not found using the buffer solutions at the high and low pH values, pH 4 and pH 8, siliconized syringe barrels may potentially be suitable for various types of injectable formulations. Previous studies have demonstrated that pH formulations may extract silicone oil from the container, due to the rapid hydrolysis of polydimethylsiloxane occurs at these conditions [3, 20]. However, toxicological assessments are still mandatory to ensure completely suitability for use and to ensure patient safety, because, in addition, as it is shown in Table 1, 12 of the 25 compounds (48%) belong to class III (either no basis to presume safety or positive indication of toxicity), according to Cramer classification, that is probably the most widely used approach for classifying and ranking chemicals according to their expected level of oral systemic toxicity, which is based on Quantitative StructureActivity Relationships (QSAR).

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4. Conclusions A general method for the determination of the extractables profile for PFS’s and their corresponding stoppers is proposed, based on liquid-liquid extraction followed by UHPLC-MS, while plunger stoppers were analysed according to ISO 10993-12:2012. The extracted compounds were identified based on chromatographic and MS data and spectral library searching. MS/MS fragmentation patterns of the compounds were also used to obtain information about the relevant structures. The established method shows advantages in sensitivity and analytical speed. Various solvents, including a mixture 2-propanol:water and buffer solutions at 3 pH conditions (4, 7 and 8) were investigated. 25 compounds in total were identified, 60% of those were found in the plunger stoppers. Brand B1 showed a higher number of extractables (72%) compared to brand B2 (48%). The method has been successfully applied to determine the extractable profile of two different brands of syringes, including the stoppers.

ACKNOWLEDGEMENTS This publication has emanated from research conducted with the financial support of Science Foundation Ireland (SFI) under Grant Numbers SFI/12/RC/2275 and SFI/13/SP SSPC/I2893 which is co-funded under the European Regional Development Fund. The authors would like to thank Amgros I/S (Denmark) for supplying syringes that were tested in this study.

DECLARATION OF INTEREST NDG and JB report no conflicts of interest. IL is an employee of Amfros I/S. The authors alone are responsible for the content and writing of this article.

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5. References [1] J. Jezek, N.J. Darton, B.K. Derham, N. Royle, I. Simpson, Biopharmaceutical formulations for prefilled delivery devices, Expert Opin. Drug Delivery 10(6) (2013) 811-828. [2] N.S. Kale, A. Kazi, S.S. Kale, Drug prefilled non-reusable syringes as drug-device, World J. Pharm. Sci. 3(10) (2015) 2095-2110. [3] R.G. Ingle, A.S. Agarwal, Pre-filled syringe – a ready-to-use drug delivery system: a review, Expert Opin. Drug Delivery 11(9) (2014) 1391-1399. [4] D.R. Jenke, Extractables and leachables considerations for prefilled syringes, Expert Opin. Drug Delivery 11(10) (2014) 1591-1600. [5] I. Markovic, Risk management strategies for safety qualification of extractable and leachable substances in therapeutic biologic protein products, Am. Pharm. Rev. 12(4) (2009) 96-101. [6] Y. Nashed-Samuel, D. Liu, K. Fujimori, L. Perez, H. Lee, Extractable and leachable implications on biological products in prefilled syringes, Am. Pharm. Rev. 14(1) (2011) 74=80. [7] R. Colton, Recommendations for extractables and leachables testing, BioProcess Int. 5(11) (2007) 36-44. [8] W. Ding, Determination of Extractables and Leachables for single use systems, Chem. Ing. Tech. 85(1-2) (2013) 186-196. [9] K. Boven, S. Stryker, J. Knight, A. Thomas, M. van Regenmortel, D.M. Kemeny, D. Power, J. Rossert, N. Casadevall, The increased incidence of pure red cell aplasia with an Eprex formulation in uncoated rubber stopper syringes, Kidney Int. 67(6) (2005) 2346-2353. [10] B. Sharma , F. Bader, T. Templeman, P. Lisi, M. Ryan, G. Heavner, Technical investigations into the cause of the increased incidence of antibody-mediated pure red cell aplasia associated with Eprex, Eur. J. Hosp. Pharm. 5 (2005) 86-91. [11] D. Liu, Y. Nashed-Samuel, P. Bondarenko, D. Brems, D. Ren, Interactions between therapeutic proteins and acrylic acid leachable, PDA J. Pharm. Sci. Technol. 66(1) (2012) 12-19. [12] A. Rosenberg, Effects of protein aggregates: an immunologic perspective, AAPS PharmSciTech 8(3) (2006) E501-507.

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[13] D. Jenke, A. Odufu, T. Couch, M. Chacko, S. Strathmann, E. Edgcomb, Evaluation of the general solution compatibility of polymer materials used in medical devices such as syringes, PDA J. Pharm. Sci. Technol. 66(4) (2012) 286-306. [14] D. Ball, D. Norwood, L. Nagao, Utility and application of analytical and safety thresholds for the evaluation of extractables and leachables in drug products, Am. Pharm. Rev. 10(5) (2007) 16-21. [15] D. Ball, D. Norwood, C. Stults, L. Nagao, Leachables and extractables handbook. Safety evaluation, qualification, and best practices applied to inhalation drug products, J. Wiley and Sons, NY, USA, 2012. [16] T. McIlvaine, A buffer solution for colorimetric comparison, J. Biol. Chem. 49 (1921) 183-186. [17] R.G. Strickley, Solubilizing excipients in oral and injectable formulations, Pharm. Res. 21(2) (2004) 201-230. [18] F. Zhang, A. Chang, K. Karaisz, R. Feng, J. Cai, Structural identification of extractables from rubber closures used for pre-filled semisolid drug applicator by chromatography, mass spectrometry, and organic synthesis, J. Pharm. Biomed. Anal. 34(5) (2004) 841-849. [19] J.R. Roy, S. Chakraborty, T.R. Chakraborty, Estrogen-like endocrine disrupting chemicals affecting puberty in humans - A review, Med. Sci. Monit. 15(6) (2009) RA137-145. [20] D. Shah, J. Cronin, M. Chacko, A. Gillum, Impact of formulation and processing parameters on silicone extraction from cyclic olefin copolymer (COC) syringes, PDA J. Pharm. Sci. Technol. 65 (2011) 109-115.

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Figure captions

Figure 1 General design of a Prefilled syringe. (A) Prefilled syringe components. (B) Design of plungers for each one of the tested brands of syringes.

Figure 2 Distribution of extracted compounds per extraction solvent. (A) Stoppers; and assembled syringes from Brand 1, (B), and Brand 2 (C). IPA = 2-propanol.

Figure 3 Total number of compounds found in assembled syringes per extraction solvent. For Brand 1, (A), and Brand 2 (B). IPA = 2-propanol.

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19

20

21

Table 1. Non-volatile extractables profile of the studied pre-filled syringes mass Name (ppm)

Formula

Neutral mass

1 C7H13NO2

143.0947

1.37

2-(Dimethylamino)ethyl acrylate

2439-35-2

2 C11H20O2

184.1463

1.20

10-Undecenoic acid

112-38-9

3 C12H14O4

222.0894

2.26

Diethyl Terephthalate/Diethyl phthalate

120-61-6/ 84-66-2

4 C11H12O3Si

220.0556

0.26

5-(Trimethylsilyl)-2-benzofuran-1,3-dione

18019-71-1

5 C8H22O3Si2

222.1107

2.24

1,3-Diethoxy-1,1,3,3-tetramethyldisiloxane

18420-09-2

6 C50H66O8

794.4758

1.71

Ethylene bis[3,3-bis[3-(1,1-dimethylethyl)-4-hydroxyphenyl]butanoate]

32509-66-3

7 C42H63O4P

662.4464

0.93

Tris(2,4-di-tert- butylphenyl)phosphate

95906-11-9

8 C9H10

118.0788

0.22

-methylstyrene

98-83-9

9 C19H32O6

356.4538

0.76

1,6-Hexanediol, tri-ethoxylated, diacrylate

-----

10 C32H66O17

722.4300

0.42

PEG n16

25322-68-4

11 C20H27O4P

362.1640

1.93

2-Ethylhexyldiphenyl phosphate

1241-94-7

12 C16H24O3

264.1726

0.05

Methyl 3,5-di-tert-butyl-4-hydroxybenzoate

2511-22-0

13 C8H22O3Si2

222.1108

2.24

1,3-Diethoxy-1,1,3,3-tetramethyldisiloxane

18420-09-2

14 C18H39O3P

334.2637

2.16

Octadecylphosphonic acid

4724-47-4

15 C20H21NO2

307.1565

0.25

4-Cyanophenyl 4-hexylbenzoate

50795-85-6

16 C22H43NO

337.3345

1.22

Erucamide

112-84-5

Annotation: CAS

Cramer Classification I

Source

Brand

Stoppers / IPA extracts

S1, B1

Stoppers / IPA extracts

S1, B1

Stoppers / IPA extracts

S1, B1

Stoppers / IPA extracts

S1, B1

Stoppers

S1

III

Stoppers pH 7 extracts

S1 B2

III

All conditions pH 7 extracts

B1 B2

Stoppers All conditions IPA + pH 7 extracts

S1 B1 B2

Stoppers All conditions IPA + pH 7 extracts

S1 B1 B2

Stoppers All conditions IPA + pH 7 extracts

S1 B1 B2

Stoppers / IPA extracts

S1, B1

Buffers pH 4 and 10 IPA extracts

B1 B2

Stoppers

S1

All conditions

B1

Stoppers IPA + pH 7 extracts

S1 B2

Stoppers All conditions pH extracts

S1 B1 B2

I I III III

I

I

I III II III III III III

(2)

(1)

22

17 C20H44N2O

328.3454

0.87

1-((2-Aminoethyl)amino)octadecan-2-ol

58436-15-0

18 C14H42O5Si6

458.1648

0.00

Tetradecamethylhexasiloxane

107-52-8

19 C7H4N5

144.0436

0.45

Pyrazolo[1,5-a]pyrimidine-3-carbonitrile

25939-87-1

20 C7H14O4

146.0943

0.13

Ethyl 3-ethoxypropanoate

763-69-9

21 C12H16O6

240.0998

0.20

3,4,5-trimethoxydihydrocinnamic acid

25173-72-2

22 C11H12O3

192.0786

1.22

Ethyl benzoylacetate

166104-20-7

23 C26H54O14

590.3514

2.01

PEG n13

25322-68-4

24 C34H70O18

766.4562

1.55

PEG n17

25322-68-4

25 C22H40O

320.3079

0.96

(2E,5S,9S)-5,9-Dimethyl-11-(2,6,6-trimethyl-1-cyclohexen-1-yl)-2-undecen-1-ol

-----

(1) (2)

III III III I I I I I I

All conditions pH 7 extracts

B1 B2

Stoppers

S1

Stoppers All conditions pH 7 extracts

S1 B1 B2

IPA + pH 7 extracts

B2

IPA + pH 7 extracts

B2

All conditions

B1

All conditions

B1

All conditions

B1

All conditions

B1

Brands: S1 = stoppers brand 1; B1 = assembled syringe from brand 1; B2 = assembled syringe from brand 2. All conditions = IPA + Buffers pH 4, 7, and 10.

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