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Determination of Ethylene Oxide and its Derivative in water by GC-FID

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Eli Grushka Igal Bar-Ilan

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Determination of Ethylene Oxide and its
Derivative in water by GC-FID
E.Grushka* and I.Bar-Ilan**
* Department of Inorganic and Analytical Chemistry The Hebrew University of Jerusalem, Israel.
** Department of Analytical Chemistry, MIGAL-Galilee Technologic Center, Kiryat-Shmona, Israel.

Abstract
Ethylene Oxide (EO) is used for sterilization of medical devices. The British
Standard (BS) defines the maximum allowable residues for EO, ethylene
chlorohydrine (ECH) and ethylene glycol (EG), for each individual medical device.
The method of choice analyzing for these residues is by GC. The current GC methods
requires two separate chromatographic runs, one for EO and the other for EG and
ECH. In addition, in the existing method the life-time of the columns is very short.
The work presented here describes a new gas chromatographic method to determine
all three components in aqueous solutions in one run. The life-time of the columns in
this method is of conventional duration.

Introduction
Determination of Ethylene Oxide (EO) and its derivatives Ethylene
Glycol (EG) and Ethylene Chlorohydrin (ECH) in medical devices is of
major concern to the health care professionals1 (public health). Gaseous
EO is used in medical products sterilization and it is important to ensure
that minimal levels of EO EG and ECH are found to minimize risk for the
patient2.
Many analytical methods for EO and its derivatives have been
described and reviewed in the literature3. All the recommended analytical
gas chromatography methods available in the standard monographs1,5,6
describe a packed column separation of these residues in aqueous
solutions. These methods usually require two separate analysis in order to
determine EO EG and ECH in short-live columns. Furthermore, the
AAMI suggests specifically separate analyses for EO5 and for EG and
ECH6.
Danielson4 first described (1990) a capillary column separation of
the three materials. The separation was done on two DB-WAX columns,
a cross-linked and bonded polyethylene glycol (PEG) on fused silica
capillary column. These columns are of high polarities and they are less
stable, less robust and have lower temperature limits than most
polysiloxanes. They exhibit shorter lifetimes and are more susceptible to
damage upon over heating or exposure to oxygen7. Polyethylene glycol
stationary phases must be liquids under GC temperature conditions.

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 1
 In the present study a detailed one-step GC analytical method for
the analysis of EO and its derivative EG and ECH in aqueous solution is
described, using a capillary column coate with dimethylpolysiloxane.
This column is non-polar and shows an excellent thermal stability of the
stationary phase. This non-polar phase is less susceptible to oxidation and
hydrolysis than phases incorporating more polar functional groups8.
Thus, the column lifetime is greatly increased.
In addition, this paper also presents the use of Solid Phase
MicroExtraction (SPME) to determine EO, ECH & EG in aqueous
solutions. SPME is used here in the headspace (HS) gas phase as well as
in the aqueous phase.

Experimental
Materials. Double Distilled Water (DDW) after 0.45µm filtration was
used. Ethylene Oxide was obtained from AirGas. Ethylene Glycol and
Ethylene Chlorohydrine were manufactured by Merck.
Standard solutions. EO from the standard gas cylinder passed through
septum with a hypodermic needle to a 30ml serum bottle. Polyvinyl
chloride tubing was connected to the vent needle to bubble EO through a
filtered DDW in a beaker. The additional weight to the DDW was
calculate to receive the EO concentration of the stock solution
(approximately 1000mg(EO)/l). 100mg of EG and ECH were transferred
to 100ml volumetric flasks and diluted to volume with DDW.
Suitable aliquots of the stock solutions were transferred to a 25ml
volumetric flask and diluted to volume. Adding appropriate volumes of
the standards stocks solution gives the calibration serial solutions.
Apparatus. A HP 5890A gas chromatograph with FID equipped with
HP 7673 autosampler and a split/splitless injector was used. The
chromatography integration is achieved using a Pentium computer
equipped with HP-ChemStation chromatography software. Helium was
the carrier gas at 1.2 ml/min (Helium velocity of 21.6 cm/sec) and it
produces a column head pressure of 6 psi at 38°C. Nitrogen was the
auxiliary gas at 30 ml/min. The septum purge gas was Helium at 3
ml/min. The injection volume was 0.5 µl (using 5 µl autosampler
syringe), at split ration of 18:1 (vent flow/column flow). The injector
temperature was 200°C and the detector temperature was 300°C. Bonded
100% dimethylpolysiloxane fused-silica capillary column was used
(Quadrex 007-1 series). The column length was 30 m with i.d. of 0.32
mm, and films thickness of 1.0 µm. This stationary phase is a non-polar
phase, which separates compounds according to boiling point, with
excellent efficiency and thermal stability (column max temperature of

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 2
350°C). The column temperature was 38°C for 0 min then it rose to
100°C at 5°C/min.
Direct Injection. Direct injections carry out with 5µl autosampler
syringe, after 3 washes with water and 3 washes in the sample solution.
The injection volume was 0.5 µl .
SPME apparatus. A manual Solid Phase MicroExtraction (SPME)
fiber holder with 100 µm polydimethylsiloxane (PDMS) SPME resin
coated fibers protected in SS syringe needle was used. The SPME was
obtained from Supelco (Bellefonte, PA). The fibers were conditioned at
200°C for 3 min.
Procedure. The standards solution was put in a 2 ml GC autosampler
vial sealed with aluminum seal with PTFE-faced red rubber septum. The
standards volume was 1 ml, which leaves 1ml air Headspace. For
Headspace analysis, 2mm of the SPME syringe penetrated through the
vial septum, then the PDMS fiber was exposed to the vial headspace for
the desired time. At the end of the adsorption the fiber was retracted to
the SS SPME syringe needle, and the SPME holder was taken out of the
vial for GC injection.

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 3
EO curve – 5 to 25 ppm (mg/l) in DDW
0.5 µl water automatic 18:1 Split injection at 200°C.

25ppm

15ppm

10ppm

5ppm

EO curve – Direct 0.5 µl water Injection (18:1 split injection)

LOD ~ 1 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 4
EO SPME curve – 5 to 25 ppm
Headspace (HS) – 15 min exposure
1 min desorption at 200°C; splitless injection

EO

25 ppm EO in DDW

15 ppm EO in DDW

10 ppm EO in DDW

5 ppm EO in

EO curve – SPME Headspace

SPME LOD ~ 5 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 5
ECH curve – 5 to 25 ppm (mg/l) in DDW
0.5 µl water automatic 18:1 Split injection at 200°C.

25 ppm

15 ppm
10 ppm
5 ppm

CH curve – Direct 0.5 µl water Injection (18:1 split injection)

SPME LOD ~ 1 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 6
ECH SPME curve – 5 to 25 ppm
Headspace (HS) – 15 min exposure
1 min desorption at 200°C; splitless injection

ECH 25 ppm

15 ppm

10 ppm

5 ppm

ECH curve – SPME Headspace

SPME LOD ~ 5 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 7
EG curve –25 to 150 ppm (mg/l) in DDW
0.5 µl water automatic 18:1 Split injection at 200°C.

150 ppm

100 ppm
50 ppm

25 ppm

EG curve – Direct 0.5 µl water Injection (18:1 split injection)

LOD ~ 20 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 8
EG SPME curve – 25 to 150 ppm
Headspace (HS) – 15 min exposure
1 min desorption at 200°C; splitless injection

EG Not detectable

SPME LOD – not detectable

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 9
EO, ECH & EG mix - GC-FID analysis.
Calibration curve
5 to 25 ppm (mg/l) standards in DDW
0.5 µl water automatic 18:1 Split injection at 200°C.

EG (4.13min)
25-150 ppm

EO (1.40min)
5-25 ppm
ECH (3.27min)
5-25 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 10
 EO (1.40min)
5-25 ppm

ECH (3.27min)
5-25 ppm

EG (4.13min)
25-150 ppm

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 11
HeadSpace (HS) SPME - GC-FID Analysis.
Calibration curves of EO, ECH & EG mix
Headspace (HS) – 15 min exposure
1 min desorption at 200°C; splitless injection

EO
5-25ppm ECH
5-25ppm
SPME Impurities

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 12
Immersion SPME - GC-FID Analysis.
Calibration curves-EO, ECH & EG mix
Water Immersion – 15 min exposure
1 min desorption at 200°C; splitless injection

EO 5- ECH 5-
25ppm 25ppm

EO curve

ECH curve

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 13
Immersion SPME - GC-FID Analysis.
Mix of 5ppm EO, 5ppm ECH & 25ppm EG
Water Immersion – 15 to 60 min exposure
1 min desorption at 200°C; splitless injection

EO curve

ECH curve

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 14
 EO Immersion

Immersion time (min)

ECH Immersion

Immersion time (min)

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 15
RESULTS AND DISCUSSION

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 16
 LITERATURE CITED

1. Biological evaluation of medical devices, part 7 – Ethylene

oxide sterilization residuals; BS EN ISO 10993-7: 1996.
2. Medical devices-Validation and routine control of ethylene
oxide sterilization; ISO 11135: 1994.
3. Biological evaluation of medical devices, part 7 – Ethylene
oxide sterilization residuals; Annex F; BS EN ISO 10993-7: 1996.
4. Danielson, J.W., Snell, R.P. and Oxborrow, G.S. Detection and
quantification of ethylene oxide, 2-chloroethanol, and ethylene glycol
with capillary gas chromatography. J.Chromatogr.Sci.28 1990; pp.97-
101.
5. Determining Residual Ethylene Oxide in Medical Devices,
ANSI/AAMI ST29-1988.
6. Determining Residual Ethylene Chlorohydrin and Ethylene
Glycol in Medical Devices, ANSI/AAMI ST30-1989.
7. J&W Scientific Incorporated 1998 catalog, Technical reference
& cookbook.
8. Quadrex Corporation 1999 Gas Chromatography Products –
Buyers Guide.

EO, ECH & EG – GC/FID Poster

Water & SPME Injections Page No. 17

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