(M1430-08-50) Leachable Analysis of Ethylene Vinyl Acetate Infusion Bags by GC-FID-MS and LC-UV-MS

Purpose: Ethylene Vinyl Acetate Infusion Bags has emerged as the preferred alternative material to replace traditional polyvinyl chloride (PVC) bags. It is important to detect and identify substances that can potentially leach into the parenteral drug product formulation from Ethylene Vinyl Acetate Infusion Bags as a result of direct contact with the formulation.

Methods: A GC-FID-MS method was used for volatile leachable analysis, with FID data for peak detection, and MS data for identification. The GC method utilized an Agilent DB-5MS, 30 m×0.25 mm ID×0.25 µm GC column, with helium as the carrier gas at 2 mL/min, inlet temperature 280 ºC, and injection volume 1 µL. Column temperature program: 40 ºC hold 1 min, ramp temperature at 10 ºC /min to 280 ºC, hold for 2.0 min., ramp temperature at 15 ºC /min to 310 ºC, hold for 5.0 min. Total Run Time: 34 min. FID detector: temperature 325 ºC. MS detector: EI+ ionization mode, with a scan range of 33-800 Daltons. Butylated hydroxytoluene (BHT) was used as the surrogate reference standard. A LC-UV-MS method was used for non-volatile organic leachable analysis, with UV data for peak detection and MS data for identification. The LC method utilized an Acquity BEH-C8, 1.7 µm, 50×2.1 mm UPLC column with a flow rate of 0.3 mL/min and injection volume of 10 µL. MPA: 10 mM Ammonium Acetate in water, MPB: Acetonitrile. A mobile phase gradient was applied from MPB 5% to 95%. Total Run Time: 23 min. UV detector: 230 nm. MS detector: EI +&- ionization mode, with a scan range of 100-1200 Daltons. Bisphenol A (BPA) was used as the surrogate reference standard.

Results: The analytical screening methods used for the leachable analysis were qualified prior to analyzing the sample. Specificity, linearity, accuracy, precision and LOQ were assessed. Correlation coefficient (R) of 0.99~1.00 for the linear regression was obtained for both the GC-FID and LC-UV methods. For the GC method, accuracy (spiking recovery) in the range of 95~104% was obtained with a %RSD of 3~4%. For the LC method, accuracy in the range of 87~112% was obtained with a %RSD of 0~3%. LOQ of 0.2 µg/mL was obtained for both the GC and LC methods, which met the sensitivity requirement for leachable analysis. The Analytical Exposure Threshold (AET) for IV bags was calculated as 0.2 µg/mL based on a Safety Concern Threshold (SCT) of 1.5 µg/day and Concentration Factor of 2. A 6-Months stability sample at accelerated storage conditions of a parenteral drug product stored in infusion bags was pulled and analyzed. A drug product sample without exposure to the infusion bag was used as negative control. One volatile leachable peak at 8.2 min was observed by GC-FID and its mass was m/z 119. This leachable was identified as 3-methylbenzaldehyde, CAS#620-23-5, using a mass spectrum library search (Figure 1). Its concentration was measured as 0.50 µg/mL corresponding to a daily intake level of 3.8 µg/day, below its Permitted Daily Exposure (PDE) of 5000 µg/day, which was obtained by assessment with the QSAR Toxicological Toolbox. One non-volatile leachable peak at 6.1 min was observed by LC-UV and its mass was m/z 311 (Figure 2). This leachable was identified as bis(2-Ethoxyethyl)phthalate, CAS#605-54-9 which is used as a plasticizer during manufacturing of the infusion bags to increase flexibility and durability. Its concentration was measured as 0.88 µg/mL corresponding to a daily intake level of 6.6 µg/day, below its Permitted Daily Exposure (PDE) of 2500 µg/day, which was obtained by assessment with the QSAR Toxicological Toolbox.

Conclusion: A GC-FID-MS method and a LC-UV-MS method were qualified for screening, quantitation and identification of potential substances leached from Vinyl Acetate Infusion Bags. The two methods were used for analysis of 6-month parenteral stability samples stored in the infusion bags at accelerated conditions. 3-methylbenzaldehyde was found as a volatile leachable substance at 0.50 µg/mL. Bis(2-Ethoxyethyl) phthalate was found as a non-volatile leachable substance at 6.6 µg/mL. Both of the leachable substances were below their Permitted Daily Exposure level per toxicological assessment.

Figure 1. GCMS Spectrum with Structure

Figure 2. LCMS Spectrum with Structure