This study aims to identify and quantify extractables from IV bags using different solvent extractions and analytical techniques. Commercially available IV bags were extracted with various solvents and the extracts were analyzed using headspace GC-MS, liquid injection GC-MS, and LC-MS. The study found cyclohexanone in all extracts and IPA migration between bags incubated together. Modifications like separating bags and using aluminum foil barriers were applied to address challenges. The IPA/water extract showed the highest extractable levels, with bis(2-ethylhexyl) phthalate being the primary extractable compound identified.

1. PURPOSE
The objective of this study is to perform an extractable
study on IV bags using different solvent systems, such
as acidified water, alkaline water, PBS buffer, and organic/
aqueous solvent mixtures to bracket and mimic pH values,
ionic strength and hydrophobicity of common IV solutions. In
order to obtain a comprehensive extractable profile, multiple
analytical techniques were used to identify and quantify the
extractables, including Headspace (HS)-GC-MS/FID analysis
for volatile organic compounds, Liquid-Injection GC-MS/FID
analysis for semi-volatile organic compounds, and LC-MS/
UV analysis for non-volatile organic compounds. There are
analytical challenges associated with those analyses that will
be addressed in this presentation.
MATERIALS AND METHODS
Sample Preparation
Commercially available IV bags purchased from Amazon were
extracted following PQRI guideline using pH 3 water, pH 9
water, IPA/water (1:1), Ethanol/water (1:1), and phosphate
buffered saline (PBS). The bags filled with each extraction
solution were placed in a shaker at 50 °C for 7 days. The
extract solutions were analyzed by LC-MS, Headspace-GC-
MS, and Liquid-Injection GC-MS.
RESULTS
Headspace GC-MS Results:
Headspace GC-MS analysis was used to identify and quantify
volatile organic compounds in all extracts. Cyclohexanone
was found in all extracts, which was also shown by Liquid-
injection GC-MS analysis. The overlap of extractable profile by
Headspace-GC-MS and Liquid Injection GC-MS has shown
the completeness of the volatile and semi-volatile organic
compound profile in our method.
Interestingly, we found that IPA was shown in the all the
aqueous extracts. We suspected that the vapor of the IPA/
water extract solvent in the IV bags could migrate into the
aqueous solvents in other bags in the same shaker, illustrated
in Figure 1. We then did more studies to separate the bags
with aqueous extraction solvents and bags with organic
solvents. There was no IPA detected after the separation
of the incubation. It means that we need to separate
incubations to avoid this migration. We have done another
investigation by putting all bags in the same shaker while
covered by aluminum foil outside the IV bags. The amount
of detected IPA was significantly decreased. It provides an
alternative approach by using aluminum foil to avoid the vapor
penetration through the plastic IV bags.
GC-MS Results:
GC-MS analysis was used to identify and quantify semi-
volatile organic compounds using Thermo Single Quad ISQ
with NIST library and Thermo Orbitrap Q-Exactive (High
Resolution Mass Spec) to assist identification.
The aqueous extracts are usually solvent exchanged
with methylene chloride (DCM) before injecting into the
GC system. However, there is often emulsion or phase
separation problem during Liquid-Liquid Extraction (LLE) if
there is high amount of alcohol in the samples, such as the
Ethanol/water extract and the IPA/Water extract. We studied
the effect of solvent exchange by injecting the Ethanol/
water extract and IPA/Water extract with and without DCM
extraction. As shown in Figure 2 to Figure 5, similar profiles
were shown with and without DCM extraction for both
Ethanol/water extract and IPA/water extract. However, better
peak shape and higher concentration/recovery were observed
for extractable peaks in the samples with DCM extraction.
This suggests the importance of DCM extraction regardless
of emulsion and phase separation issues. We have also
investigated the emulsion and phase separation problems,
and found out no effect on recovery and peak shape.
The IPA/Water extract has shown the highest concentration
of extractable compounds, especially hydrophobic
compounds, comparing to other extracts (Ethanol/Water, pH
3, pH 9 and PBS). The major peak observed from all extracts
is Bis (2-ethylhexyl) phthalate (DEHP), which is the primary
plasticizer of this PVC bag. Most of extractable compounds
are phthalates, degradants from DEHP and lubricants.
ANALYTICAL CHALLENGES IN
EXTRACTABLE STUDIES OF IV BAGS
PART 1: HEADSPACE AND LIQUID INJECTION GC-MS ANALYSIS
DUJUAN LU¹, KENNETH WONG¹, KATE COMSTOCK², EKONG BASSEY², JING KONG¹, DANNY HOWER¹, JOHN SCHMELZEL², AND KELLY BERTRAND¹
1
SGS LIFE SCIENCE SERVICES, 2
THERMO FISHER SCIENTIFIC
SGS
INTRODUCTION
Intravenous (IV) therapy is commonly used in hospitals. Solutions in IV bags, such as IV drugs, blood-based products and parenteral
nutrition, are infused directly into the veins of the patients in significant amounts. Most of the IV bags are made of plastics and those
plastics additives and degradants may easily leach out into the IV solutions. As part of safety risk assessment, it is very important to identify
and quantify those extractables and leachables as they may pose safety risks to patients and/or change the efficacy of the medical products.
CONCLUSION
We have done a thorough extractable study for IV bags by
extraction with pH 3 water, pH 9 water, IPA/water (1:1),
Ethanol/water (1:1), and phosphate buffered saline (PBS) and
analyzed by Headspace-GC-MS, Liquid Injection GC-MS, and
LC/MS. We have encountered and solved several analytical
challenges during this extractable study of IV bags. During
Headspace-GC-MS analysis, there was one unexpected
finding that the vapor of the volatile organic solvents in the IV
bags could migrate into the solvents in other bags. Modified
experiments were then applied to investigate the root cause
and effectively eliminated the problem. In addition, the
importance and analytical challenges of DCM extraction with
organic extracts was studied during GC-MS analysis.
See part two for the LC-MS analysis
FIGURE 5: GC-MS CHROMATOGRAM OF IPA/WATER
EXTRACT (WITH DCM EXTRACTION)
FIGURE 4: GC-MS CHROMATOGRAM OF IPA/WATER
EXTRACT (WITHOUT DCM EXTRACTION)
FIGURE 2: GC-MS CHROMATOGRAM OF ETHANOL/
WATER EXTRACT (WITHOUT DCM EXTRACTION)
FIGURE 3: GC-MS CHROMATOGRAM OF ETHANOL/
WATER EXTRACT (WITH DCM EXTRACTION)
Headspace (HS) - Gas Chromatography (GC) Conditions
Headspace Analyzer: Agilent G1888 Headspace Sampler
Vial Oven Temperature: 80 °C
Needle Temperature: 110 °C
HS Transfer Line
Temperature:
140 °C
Thermostatting Time: 30 min
Injection Time: 1 min
Gas Chromatograph: Agilent 6890N
Column: DB-624, 30 m × 0.32 mm × 1.8 µm
Temperature Program: Start at 40 °C, Hold 1.0 min.
Ramp at 7 °C/min to 200 °C,
Then Ramp at 20 °C/min to 250 °C.
Hold 3.0 min.
Total Run Time: 29 min.
Carrier Gas: Helium
Injector mode: Split with a ratio of 1:1
Split Flow: 1.0 mL/min.
Injector Temperature: 140 °C
Flame Ionization Detector (FID)
Detection Temperature: 250 °C
Air Flow: 400 mL/min.
Hydrogen Flow: 35 mL/min.
Make-Up Gas Flow: 5.0 mL/min. Helium
Mass Spectrometry (MS)
Mass Spectrometer: Agilent 5973
Ion Mode: EI+
Scan Range: 29-300 m/z
Source Temperature: 200 °C
Transfer Line Temperature: 200 °C
Electron Energy: 70 eV
Solvent Delay: 0 min
Calibration Gas: Perfluorotributylamine (PFTBA)
Liquid-Injection GC-MS Conditions
Gas Chromatograph: Thermo Trace 1310 GC
Column: TG-5HT (30 m × 0.25 mm × 0.10 μm)
Temperature Program: Start at 40 °C, Hold 1.50 min.
Ramp at 10 °C/min. to 250°C,
Hold 0.5 min.
Ramp at 5 °C/min. to 295°C,
Hold 10 min.
Ramp at 10 °C/min. to 300°C,
Hold 1.0 min.
Total Run Time: 45 min
Carrier Gas: Helium
Carrier Gas Flow: 1.5 mL/min
Injector Temperature: PTV: Start at 65 °C, Ramp at 10 °C /sec
to 295 °C, hold for 0.5 min
Injector: splitless (for 1.0 min)
Injection Volume: 1.0 µL
Flame Ionization Detector (FID)
Detection Temperature: 325 °C
Air Flow: 400 mL/min.
Hydrogen Flow: 30 mL/min.
Make-Up Gas Flow: 21 mL/min. Helium
Mass Spectrometry (MS)
Mass Spectrometer: Thermo Single Quad ISQ
Thermo Orbitrap Q-Exactive
Ion Mode: EI+
Scan Range: 25-650 m/z
Source Temperature: 305 °C
Transfer Line Temperature: 300 °C
Electron Energy: 70 eV
Solvent Delay: 4.0 min
Calibration Gas: Perfluorotributylamine (PFTBA)
FIGURE 1: ILLUSTRATION OF IPA MIGRATION
THROUGH IV BAGS