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Communications in Soil
Science and Plant Analysis
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Automated analysis of
gases stored in Vacutainer
vials
a a
Jeffrey A. Covert , Mario Tenuta & Eric G.
a
Beauchamp
a
22135, Newton Ave, Halifax, B3L 3C1,
Canada
b
Land Resource Science , University of
Guelph , Guelph, NIG 2W1, Canada
Published online: 11 Nov 2008.

To cite this article: Jeffrey A. Covert , Mario Tenuta & Eric G. Beauchamp
(1995) Automated analysis of gases stored in Vacutainer vials ,
Communications in Soil Science and Plant Analysis, 26:17-18, 2995-3003, DOI:
10.1080/00103629509369503

To link to this article: http://dx.doi.org/10.1080/00103629509369503

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 COMMUN. SOIL SCI. PLANT ANAL., 26(17&18), 2995-3003 (1995)

AUTOMATED ANALYSIS OF GASES STORED IN VACUTAINER1

Downloaded by [University of Illinois at Urbana-Champaign] at 12:12 17 March 2015

VIALS
Jeffrey A. Covert
22135, Newton Ave., Halifax, B3L 3C1, Canada
Mario Tenuta, and Eric G. Beauchamp2
Land Resource Science, University of Guelph, Guelph, NIG 2W1, Canada

ABSTRACT: Trace gas research is often constrained by the number of samples

to be analyzed. We report here, modifications to a commercially available headspace
gas autosampler for sampling stored gas in blood collection vials (Vacutainer
Brand). The sampler was modified to: a) accommodate a 0.5-mL gastight syringe,
b) accept 3-mL Vacutainer vials, and c) ensure the syringe needle dislodged from
the vial septum. Sample volumes using the modified sampler were consistent from
vial to vial. For the determination of the nitrous oxide (N2O) concentration of a
known reference, we found a coefficient of variation of 3% with the modified
autosampler compared to 1% with manual injections. The modification of two
autosamplers has permitted analysis of 90 sample vials per day for N2O, carbon
dioxide (CO2), oxygen (O2), acetylene (C2H2), and nitrogen (N2) gases using one
gas Chromatograph with two detector systems. As a consequence, we have
increased sample numbers, reduced associated labour costs and maintained analysis
quality.

INTRODUCTION
The measurement of greenhouse gases [CO2, N2O, and methane (CH4)] has
recently been intensified because of concerns for global warming (1). Knowing the
rate of trace gas emission or consumption in soil from a wide range of land uses

1. Registered trademark of Becton Dickinson Copany.

2. Corresponding author.

2995

Copyright © 1995 by Marcel Dekker, Inc.

 2996 COVERT, TENUTA, AND BEAUCHAMP

and geographical regions, identification and modeling the major soil sources and
sinks of trace gases is possible (2).
Trace gas research is often constrained by the number of samples that can be
analyzed. The length of time and labour costs required to manually introduce gas
samples into a gas Chromatograph are particularly time and labour consuming.
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Ideally, an automated gas introduction system, should sample the headspace gas of
air-tight containers that require a small volume of stored gas. Present automated
sample introduction systems such as vacuum stream selectors are not suited to
analysis of gas in small air-tight containers. Although stream selectors may be used
to sample gas in Teflon3 or Tedlar3 sample bags, these storage bags are expensive,
require large sample volumes and are labour intensive to ensure minimal con-
tamination from previously stored gas. Pressurized-vial static headspace sampling
systems are not suitable for trace gas analysis from small air-tight containers. These
systems dilute sample gas by pressurizing the storage vial with an inert gas
resulting in lower analytical sensitivity. Furthermore, vials cannot be re-sampled
because of more dilution and occurrence of leaks in vial septa with another
pressurization. Although liquid autosamplers can sample the headspace gas of
standard autosampler vials, the vials and septa available were found not to be
air-tight and thus unreliable.
Our laboratory has found blood collection vials (e.g. Vacutainer brand; Fisher
Scientific, Nepean, Canada) to be a low cost means for short term storage (1-4
weeks) of gas samples. In addition to their low cost, blood collection vials have the
advantage of remaining airtight when sampled repeatedly, can be reused to store
another sample, and require small volumes of sample gas. We report here a set of
inexpensive modifications to Varian 8200CX AutoSamplers (Varian Canada,
Mississauga, Canada) facilitating the sampling of gas from 3 mL blood collection
vials.

MODIFICATIONS
The liquid autosampler was modified in three ways: a) installation of a gas tight
sampling syringe, b) restructuring of the sample tray to hold 3 mL Vacutainers, and
c) modification of the sampler to allow gas sampling from the thicker gas-tight septa
of Vacutainer vials.

3. Registered trademark of E.I. duPont de Nemours and Company.

 ANALYSIS OF GASES IN VACUTAINER VIALS 2997

A 0.5 mL Pressure-Lok4 Series A syringe (Supelco, Oakville, CAN) was fitted

to the B200CX AutoSampler instead of the 100 u,L SGE (Kingwood, Australia)
gas tight syringe provided with the autosampler. The SGE syringe is the only
commercially available gas tight syringe fitted to the B200CX AutoSampler. When
installed, the maximum sampling volume by this syringe was 50 ^iL due to
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limitations in the sampler's built-in control software. This volume was too small to
reliably detect near-ambient concentrations of N2O and CO2 gases in Vacutainer
vials. A second shortcoming of the syringe was the needle thickness. It was too
small to withstand the stress of insertion into a Vacutainer vial rubber septum. Both
of these problems were solved by fitting a 0.5 mL gas tight PressureLok syringe
with a 225 gauge needle to the 82OOCX AutoSamplers.
The Pressure-Lok syringe was fitted to the autosampler syringe mount by
machining the syringe barrel thus allowing the syringe mount door to close. The
plunger shaft of the syringe was extended with a metal rod (3.5 cm length, 2 mm
diameter) fitting the plunger shaft into the plunger drive (Fig. 1). Conical side port
needles 6.5 cm in total length and gauge 225 were used to replace the needle
provided with the syringe (custom order from Supelco, Oakville, CAN).
The standard sample tray was adapted for 3 mL Vacutainer vials. A plexiglass
adapter tray was constructed and screwed to the top of the original tray (Fig. 2).
The adapter consisted of two plexiglass sheets spaced 3 cm apart to provide a two
point support for the vials. Holes (1.2 cm diameter) were precisely drilled in the
plexiglass sheets to match the centers of the wells in the sample carousel. The
plexiglas sheets were secured to the carousel with screws. The entire carousel unit
was lowered 2 cm to accommodate the Vacutainer vials. This was done by lowering
the hinge mounts on the chassis of the autosampler.
Vacutainer vials have thick rubber septa providing an air-tight seal but prevent
withdrawal of the syringe needle from the vial. As a result the syringe carriage and
vial were lifted together following sampling. Although the sampler came equipped
with a metal bar extension to dislodge the vial from the syringe needle as the
syringe carriage was lifted, the carriage motor was too weak to pull the syringe
needle out of the Vacutainer septum. We solved this problem by mounting a stage
to the carrousel with steel angle brackets (Fig. 2). The stage prevented removal of
the vial from the carousel as the syringe carriage moved upward after sampling. To

4. Registered trademark of Dynalcch Precision Sampling Corporation.

 2998 COVERT, TENUTA, AND BEAUCHAMP

Plunger Driver
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Switch 1-
Switch 4 -
Plunger Shaft
Pressure-Lok
Syringe
Pneumatic
Cylinder Syringe Mount
"Syringe Carriage
Switch 3.

Carousel Unit
Switch 2

FIGURE 1. Front View of Modified 8200CX AutoSampler; a) With and b)

Without the Carousel Unit.

witch 3
•3ml Vacutainer
Top Angle-^5».
Bracket Plexiglass
Sheet
Stage Bracket-f^» ase Plexiglass
Sheet

Bottom Angle
Bracket

FIGURE 2. Side View of the Modified Carousel Unit.

 ANALYSIS OF GASES IN VACUTAINER VIALS 2999

aid the sampler motor in pulling the syringe needle out of the vial, a pneumatic
piston was attached to the syringe carriage to push against the stage.
Two 12V dc computer relays connected to four subminiature SPDT lever
switches fastened to the stage, controlled the pneumatic cylinder by two solenoid
valves (Fig. 3). We used the wash cup pneumatic piston and air solenoids for the
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rinsing of the liquid syringe system already present on the autosampler to reduce the
cost of the modifications. The relays and switches are powered by a 12V dc power
adapter (input 120V-60Hz-14W, output 12V-500 mA) plugged into a standard
120V wall outlet. A 24V dc service was available from the autosampler, however
since the switches and their power contacts are exposed to the operator, a 12V
current was used to prevent injury. The four switches are triggered by various
mechanical events of the autosampler (Table 1). Depending upon the contact of the
switches, one relay triggers an "in air" solenoid to allow pressurized gas to extend
the pneumatic cylinder and the other relay triggers an "out air" solenoid, venting the
solenoid and retracting the cylinder (Table 1 and Figure 3). Alternatively, the use of
contact switches to trigger the extension of the pneumatic cylinder can be avoided if
the control software of the sampler is programmed to extend the pneumatic
cylinder.

OPERATION
A Varian 3600 Gas Chromatograph (Varian Canada, Mississauga, Canada)
equipped with two modified 8200CX AutoSamplers, was used to measure N20,
CO2, O2, C2H2, and N2 gases stored in 3-mL Vacutainer vials. The gas Chromato-
graph included a electron capture detector for the measurement of N2O and C2H2
and a thermal conductivity detector for the measurement of CO2, O2, C2H2, and
N2. The carrier gas flow and column separation systems for each detector are
isolated and an autosampler is dedicated to each detector. The autosamplers can be
controlled individually or simultaneously.
Trace gas samples may be stored in blood collection vials for up to four weeks
if they had been first opened for several hours and evacuated with a pump or
syringe. It is critical that the vials be opened prior to collection of gas samples
containing N2O and CH4 because of inconsistent headspace contamination when
shipped from the manufacturer.
The precision of the determination of sample gas concentration using the
modified autosampler was comparable to that by manual sample introduction.
 3000 COVERT, TENUTA, AND BEAUCHAMP

In Air Solenoid Out Air Solenoid

12V dc computer
relays
•24V dc from
autosampler
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autosampler
chassis

nc = normally closed
12V de
adapter nq nc no = normally open
coil = relay coil
Switch 1 Switch 2 Switch 3 Switch 4 g = ground

FIGURE 3. Wiring Diagram for the Activation of the Pneumatic Cylinder by

Solenoid Valves.

Analysis of ten 3-mL Vacutainer vials having their headspace volume removed by
manually drawing a gas tight syringe to 0.5 mL and replaced with 0.5 mL of a 3.5
ppmv-N2O standard gas resulted in a coefficient of variation of 3% using the
modified autosampler system. The modified autosampler was set to the maximum
sample volume (corresponding to 0.19 mL) allowed by the control software. A
coefficient of variation of 1% was determined for ten manual injections of 0.5 ml
from the same standard gas placed into 250-mL glass bottles at atmospheric
pressure. Plots of peak area to gas concentration were linear for N2O (Fig. 4),
CO2,02, and N2 for typical concentration ranges with both automated and manual
sample introductions methods.
Occasionally, the syringe needle failed to penetrate a vial septum. To determine
when sampling failed, C2H2 gas (0.2 mL) was injected into each vial prior to
Downloaded by [University of Illinois at Urbana-Champaign] at 12:12 17 March 2015

ANALYSIS OF GASES IN VA(ZVTA INER VIA

TABLE 1. Control of the Pneumatic Cylinder by Autosampler Events and Purpose for Triggering Switch Closure.

Switch 1 Switch 2 Switch 3 Switch 4

Event Causing syringe carriage in syringe carriage in movement of vial to complete discharge
Switch Closure resting position injector stage upon needle of gas from syringe
exit from vial

Purpose vent solenoids to set piston to extend extend piston extends piston
G
prolong piston seal upon complete against stage, against injector port
life discharge of sample pushing needle out base, pushing needle
gas from syringe of vial septum out of injector when
switch 2 is closed
 3002 COVERT, TENUTA, AND BEAUCHAMP

7
ppmv N2O= 1.416x1 o"4 x area -0.297
6-
r 2 = 0.999
5-
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4-

10000 20000 30000 40000 50000

N P O peak area

FIGURE 4. Concentration of N O in the Headspace of Vacutainer

Vials as a Function of Peak Area Using the Modified
Autosamplcr

analysis. Since C2H2 was detected by the electron capture and thermal conductivity
detectors, the presence of an C2H2 peak during a sample run indicated successful
sampling.
Automated sampling of gas stored in Vacutainer vials resulted in minimal
sample to sample carry-over. Residual gas in the syringe was removed by program-
ming the autosampler to fully withdraw the plunger shaft for 3 min. Aeration of the
syringe barrel presumably allowed sample gas to desorb from the inner wall of the
syringe barrel. This desorbed gas together with residual sample gas in the syringe
needle was expelled as the plunger shaft returned to it's resting position.
The modification of two 8200CX AutoSamplers required three weeks of labour
for one person at regular work hours to complete and U.S.$500 in parts. The modi-
fications of the two samplers has permitted our laboratory to analyze 90 samples per
regular work day for N2O, CO2, O2, C2H2, and N2 along with several reference
standards. This system has allowed our lab to increase sample numbers, reduce
associated labour costs and maintain acceptable analytical quality.
 ANALYSIS OF GASES IN VACUTAINER VIALS 3003

REFERENCES:

1. Houghton, J.T., G.J. Jenkins, and J.J. Ephraums (eds.). 1990. Climate
Change; The IPPC Scientific Assessment. Cambridge University Press,
Cambridge, MA.
2. Kreileman, G.J.J. and A.F. Bouwman. 1994. Computing land use emissions
Downloaded by [University of Illinois at Urbana-Champaign] at 12:12 17 March 2015

of greenhouse gases. Water Air Soil Pollut. 76:231-258.