Development and Validation of A Gas Chromatography Mass Spectrometry Method For The Determination of Pcbs in Transformer Oil Samples Application On Real Samples From Botswana 2157 7064.1000116

Chromatography
Motladiile et al., J Chromatograph Separat Techniq 2011, 2:4

http://dx.doi.org/10.4172/2157-7064.1000116
Separation Techniques
Research
Article
Research
Article
Open
OpenAccess
Access
Development and Validation of a Gas Chromatography-Mass

Spectrometry Method for the Determination of PCBs in Transformer Oil
Samples-Application on Real Samples from Botswana
Saidy Motladiile1*, Habauka M Kwaambwa2 and Kwenga Sichilongo3
Department of Waste management and Pollution Control, National Environmental Laboratory, PB BO323, Gaborone, Botswana
Polytechnic of Namibia, Department of Natural Sciences, PB 13388, Windhoek, Namibia
3
The University of Botswana, Faculty of Science, PB UB 00704, Gaborone, Botswana
1
2
Abstract
A procedure in which a retention time window for the determination of polychlorinated biphenyls (PCBs)
congeners including those not presented in this study but of similar molar masses in transformer oil has been
developed and validated. This procedure was successfully applied in the determination of transformer oil samples
from Botswana for comparison with a preliminary inventory for PCB levels in transformer oil and other receptacles
suspected to contain PCBs and developed during the formulation of the Botswana National Implementation Plan
(NIP) for the sound management of persistent organic pollutants (POPs). Recoveries in the developed procedure
initially meant for 39 PCB congeners ranged from 44.530.31 to 111.934.75% uncorrected for bias, while linearities
i.e. r2 values ranged from 0.9624 to 0.9999 with method detection limits (MDLs) varying from 0.02 to 0.13 ppm (w/v).
Application of this procedure on real samples from Botswana showed that the transformer oil contained no PCBs
although the preliminary data using screening equipment and kits for indicative inventory data showed high PCB
contents in some cases of as much as 456 ppm (w/v). It is proposed here that this method be adopted in the SADC
region for confirmation of presence and determination of the levels of PCBs where they are detected in order to
improve on the preliminary inventories that were developed during the NIP development exercise for the Stockholm
Convention (SC) in these countries.
Keywords: PCBs; GC-MS; Botswana; Stockholm convention; SADC;
Investigations in many parts of the world have revealed widespread

diffusion of PCBs in the environment [3]. Between 1950 and 1980,
over 1.3 million tons of PCBs were produced worldwide [4]. Although
the manufacture of PCBs since the 1970s was largely banned, their
continued use in existing materials was not. The interest in PCBs as
environmental pollutants began in 1966 when they were catalogued as
hazardous waste [5].
to the increased awareness that persistent organic pollutants (POPs)

pose to human health and the environment, the Governing Council
of the United Nations Environment Programme (UNEP) in May
1995 requested in its decision 18/32 that an international assessment
process be undertaken of an initial list of 12 POPs (aldrin, chlordane,
1,1,1-trichloro-2,2-di(4-chlorophenyl)ethane (DDT), dieldrin, dioxins,
endrin, furans, hexachlorobenzene, heptachlor, mirex, PCBs and
toxaphene). The Council also requested that the Intergovernmental
Forum on Chemical Safety (IFCS) develop recommendations on
international action for consideration by the UNEP Governing Council
and the World Health Assembly no later than 1997 [8]. Negotiations led
to the adoption of the Stockholm Convention (SC) on POPs, which was
opened for signature at a Conference of Plenipotentiaries held from 22
to 23 May 2001 in Stockholm, Sweden (Stockholm Convention, 2004).
The SC, which entered into force in 2004, provides for the protection
of human health and the environment from POPs. The SC outlines
measures for monitoring and elimination of PCBs in transformers and
other receptacles by 2025.
Due to their physical and chemical properties, PCBs are highly

persistent in the environment and tend to accumulate progressively
in soils, plants and animals and proportionally to the trophic level in
the food chain and are classified as probably carcinogenic [6,7]. Due
*Corresponding author: Saidy Motladiile, Department of Waste management

and Pollution Control, National Environmental Laboratory, PB BO323, Gaborone,
Botswana, Tel: +267 3934479; Fax: +267 3934486; E-mail: smotladiile@gov.bw
National implementation plan
Introduction
Polychlorinated biphenyls (PCBs) are a class of persistent organic
pollutants (POPs) used as dielectric fluids because of their chemical,
physical stability and electrical insulating properties [1]. They are
compounds synthesized from the substitution of hydrogen atoms by
chlorine atoms in the biphenyl molecule as shown in Figure 1. There
are 209 possible PCB congeners that have only been synthesized under
laboratory conditions [2].
2'
4'
4
m
ReceivedDecember 02, 2011; Accepted January 03, 2012; Published January

06, 2012
3'
6'
5' n
m + n are chlorine atoms between 1 and 10 that can attach to the biphenyl
ring in the positions indicated
Figure 1: General structure of a PCB.
J Chromatograph Separat Techniq

ISSN:2157-7064 JCGST, an open access journal
Citation: Motladiile S, Kwaambwa HM, Sichilongo K (2011) Development

and Validation of a Gas Chromatography-Mass Spectrometry Method for the
Determination of PCBs in Transformer Oil Samples-Application on Real Samples
from Botswana. J Chromatograph Separat Techniq 2:116. doi:10.4172/21577064.1000116
Copyright: 2011 Motladiile S, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited.
Volume 2 Issue 4 1000116
Citation: Motladiile S, Kwaambwa HM, Sichilongo K (2011) Development and Validation of a Gas Chromatography-Mass Spectrometry Method for
the Determination of PCBs in Transformer Oil Samples-Application on Real Samples from Botswana. J Chromatograph Separat Techniq
2:116. doi:10.4172/2157-7064.1000116
Page 2 of 8
Botswana became party to the SC on the 28th of October 2002. As
such Botswana is obliged to implement requirements of the SC. The
Botswana National implementation plan (NIP) for POPs, showed that
there were about 500 pieces of equipment containing PCBs, thus a total
oil of about 16,520 cubic metres suspected to contain PCBs in equipment
in use [9]. Indicative results of the preliminary screening exercise used
for the preparation of the NIP in Botswana for the presence of PCBs
in transformer oil and other receptacles, were done using analytical
equipment and test kits such as the CLOR- N- Oil 50 that would only
provide general tests and therefore preliminary indicative data. This
was the same in most other SADC state parties to the SC.
While several methods [10-20] have been used for the
determination of PCBs in transformer oil and other matrices, suffice
to mention that adaptability of these methods depends on a particular
region or geographical location of the analysts. This is mainly due
to the cost associated with acquisition of instruments, their service/
maintenance and affordability of reagents and other consumables.
Some examples of extraction methods that have been employed in
the determination of PCBs in transformer oil are solvent extraction
by shaking, solvent extraction in a soxhlet or Soxtec apparatus,
focused microwave-assisted solvent extraction (FMASE), in a soxhlet
apparatus, ultra sonication extraction (USE), microwave-accelerated
extraction (MAE), accelerated solvent extraction (ASE; also known as
PFE or PLE), supercritical fluid extraction (SFE), membrane extraction
techniques and solid-based extraction techniques [21]. A recent study
[22] compared the efficiencies of extraction of PCBs in food with
different sorbents including florisil but did not include the sulfoxide
sorbent. This study employed electron capture detector (ECD) for
detection. The main detectors that have been used are the ECD and
mass spectrometry employing various mass analysers such as the
quadrupole ion trap (QIT), quadrupole and the time of flight in the
electron ionization mode [23].
The study reported here focused on developing a simple, robust,
selective and sensitive retention time window that can be used to
determine the concentrations of PCBs in transformer oil and other
receptacles suspected to contain PCBs in the Southern African
Development Conference (SADC) region. Eventually, all SADC
countries could compare their preliminary PCB inventories in their
NIPs with the data adduced from scientific determination such as this
one. Samples in this study were taken from some sites in Botswana and
determined for the presence of PCBs using the method developed in
this study.
Experimental
Reagents and chemicals
HPLC grade hexane, acetone and sulphuric acid were purchased
from Merck South Africa (SA). The Solid phase extraction (SPE)
cartridges, Florisil sep-pak vac 20 cc, 2 g, were purchased from Waters
Ireland. Supelclean Sulfoxide sep-pak vac 12 cc, 3 g were purchased
from Supelco Analytical USA. The PCB standards including the
analytical standard containing 39 PCB congener mix, C-CS-01, 10
g/mL were all purchased from AccuStandard Inc. (New Haven, CT,
USA).
Apparatus
The Agilent 5975 C inert XL EI/CI MSD and the Waters GCT
Premier GC-MS system were used. Solid phase extraction (SPE) was
carried out on an IST Vacc master with 10 sample processing manifold
acquired from Biotage, Charlotte, NC, USA. This is a vacuum manifold

(Vacc Master) with capabilities of adjusting the pressure to attain the
desired flow rate.
Chromatographic conditions
The optimization of the Gas Chromatography (GC) separation
parameters for the PCBs was done using an Agilent GC 6890 equipped
to an Agilent 7683 B series injector. The detector used was a Time of
Flight (TOF) GCT Premier Micro-Mass mass spectrometer. The system
was operated using the Masslynx Version 4.1 acquisition software. The
experimental protocol was designed in such a way that the optimized
separation parameters would be transferred to another Agilent GC
system equipped with a similar injector. The column used was a DB5MS (methylpolysiloxane, 5% phenyl groups), 30 m x 0.25 mm i.d., and
0.25 m thickness. After optimization of the separation parameters,
mass spectrometry parameters were optimized using an Agilent GC
7890A coupled to an Agilent 5975 C inert XL EI/CI MSD with a Triple
axis detector manufactured by Agilent technologies, USA. The rest of
the experiments including method detection limits were determined
using this instrument. The instrument has the capability of being
operated in both full and selected ion monitoring (SIM) scan modes.
On this system, an HP5-MS column (methylpolysiloxane, 5% phenyl
groups), 30m x 0.25mm i.d., 0.25m thickness which is equivalent to
the DB5-MS column used on the GCT Premier was used. Helium was
used as a carrier gas at a flow rate of 1 mL/min. 1 L of sample/standard
solutions was injected. The GC optimized oven program was as follows:
The column was held at 80C for 2 min and then ramped at 10C/min
up to 270C. The injector was held at an optimized temperature of
270C.
Mass spectrometry
The mass spectrometer was operated in the electron ionization (EI)
mode at electron energy of 70 electron volts (eV) and a scan range of 50500 m/z. The emission current set by the instrument autotune function
was 34 A. The source and analyzer temperatures were optimized and
held at 230 and 150C, respectively, throughout. The SIM and full
scan modes of operation were compared and the best mode used in
the quantification of real samples. The acquisition software on this
system was MSD Chemstation. Data analysis was accomplished by a
combination of Productivity Chemstation and the Automated Mass
Spectral Deconvolution and Identification System (AMDIS) by the
National Institute for Standards and Technology (NIST).
Sample collection and storage

The sample used for method validation was one of the transformer
oils collected during an inventory exercise conducted in collaboration
with the Botswana Department of Waste Management and Pollution
Control (DWMPC) in November 2010. Samples were collected in
glass sampling bottles initially rinsed with n-hexane 95% UL (HPLC
grade). 25 transformer oil samples were collected and stored in ice and
transported to the laboratory for storage in the cold room maintained
at 4C before extraction and analysis. In this study, six out of twenty
five samples were analyzed. Sample collection from the transformers
entailed opening the drainage tap on the side of the transformer and
draining the oil into the glass sample bottle. Some transformers did
not have a drainage tap and these were sampled using a glass pipette
previously rinsed in hexane. The coordinates for each sample were
recorded for future sampling and analysis except for the transformers,
which were located in, protected areas. These transformers coordinates
were not recorded. The coordinates are shown in Table 1.
2:116. doi:10.4172/2157-7064.1000116
Page 3 of 8
Sample Code
Coordinate
Sample Type
Transformer
Fabrication
Date
Sample
Collection
Date
SSKA 004
E. 24o.3337.1
Transformer oil
S. 25o.5522.6
1983
04/11/2010
SSKA 009
E. 24o.3337.1
Transformer oil
S. 25o.5522.6
1983
04/11/2010
SSKA 024
E. 24o.3239.8
Transformer oil
S. 25o.5588.8
1990
04/11/2010
SSKB 002
**
Transformer oil
1989
05/11/2010
Glen valley 006
**
Transformer oil
n/a*
05/11/2010
E. 24o.3117.9
Transformer oil
S. 23o.5918.5
1998
05/11/2010
Mmamashia 003
repeating the process until the acid layer was no longer coloured.
The concentrated sulphuric acid charred most of the transformer oil
resulting in a crude black mixture of acid and oil. After acid treatment,
the treated sample was washed twice with 5 mL fractions of de-ionised
water to remove any traces of acid. Each 5 mL wash was done in a
minute. The extract was passed through anhydrous sodium sulphate
analytical reagent (AR) quality to remove moisture. The sample was
ready for SPE clean up.
SPE Clean up and extraction: A comparison of the PCB extraction
efficiencies was done using two types of SPE cartridges. The Supelco
supelclean sulfoxide 3 g, 6 mL cartridges and the Waters florisil, 2 g
sorbent, 12 cc.
*n/a- not available

** Transformers in security protected areas
Table 1: Coordinates for some transformer samples collected during the inventory
exercise.
Sample preparation
Samples: The samples were diluted in the ratio of 1:500 v/v
transformer oil samples to n-hexane. 2 mL of the diluted transformer
oil sample was cleaned up with 5 mL concentrated sulphuric acid.
This was adopted from the USEPA Method 3665A and slightly
modified by adjusting the volumes of sulfuric acid added. The method
involved shaking the mixture with concentrated sulphuric acid for a
minute, allowing the phases to separate, removing the acid layer and
Retention
Time (min)
% Match Factor
(simple)
SIM ion
(m/z)
2-Chlorobiphenyl
10.30
97
188
3-Chlorobiphenyl
11.45
98
4-Chlorobiphenyl
11.57
2,2'-Dichlorobiphenyl
Supelclean Sulfoxide SPE cartridges: The SPE normal procedure

of conditioning, loading, washing and elution was followed. The
conditioning was accomplished by eluting 10 mL of acetone to remove
residual moisture from the supelclean sulfoxide cartridges. This was
followed by adding 20 mL of n-hexane to equilibrate the cartridges.
The pre-treated sample was loaded onto the cartridge and washed with
5.5 mL of n-hexane. Elution was done with 13 mL of n-hexane. The
eluate was concentrated to 1 mL by gentle nitrogen blow. The samples
extracts were transferred into GC vials ready for analysis.
Florisil SPE cartridges: Conditioning for the florisil cartridges was
done by passing 8 mL of hexane through the cartridge. The solvent was
Retention
Time (min)
SIM ion
(m/z)
% Match Factor
(simple)
2,2',3,3',4-Pentachlorobiphenyl
18.18
326
98
188
2,2',3,4,5'-Pentachlorobiphenyl
18.64
326
97
96
188
18.83
326
97
12.07
99
222
2,3,3',4',6-Pentachlorobiphenyl
19.06
326
98
12.79
100
222
2,2',3,4,4',5'-Hexachlorobiphenyl
19.20
360
96
2,4'-Dichorobiphenyl
13.06
100
222
2,2',3,4',5,5'-Hexachlorobiphenyl
19.68
360
96
2,5 -Dichlorobiphenyl
13.23
100
222
2,2',3,4',5,6-Hexachlorobiphenyl
19.82
360
87
2,2',3-Trichlorobiphenyl
13.69
99
256
2,2',3,3',4,5,6-Heptachlorobiphenyl
20.06
394
96
14.27
100
256
20.36
360
96
14.72
99
256
2,2',3,3',4,5,6'-Heptachlorobiphenyl
20.67
394
94
2,3,4'- Trichlorobiphenyl
15.26
99
256
2,2',3,3'4',5,6-Heptachlorobiphenyl
21.22
394
94
15.44
99
256
21.34
394
94
2,4,4'-Trichlorobiphenyl
15.82
99
256
2,2',3,4,4',5,5'-Heptachlorobiphenyl
21.60
394
94
2,2',3,5'-Tetrachlorobiphenyl
16.20
98
292
2,2',3,4',5,5',6-Heptachlorobiphenyl
21.93
394
97
16.63
98
292
2,2',3,3',4,4'5,5'-Octachlorobiphenyl
22.89
430
94
2,3,3',4'-Tetrachlorobiphenyl
16.86
98
292
2,2',3,3',4,4'5,6-Octachlorobiphenyl
23.08
430
96
17.23
98
292
2,2',3,3',4,5,5',6'-Octachlorobiphenyl
24.05
430
94
2,3',4,5-Tetrachlorobiphenyl
17.46
98
292
2,2',3,4,4',5,5',6-Octachlorobiphenyl
24.80
430
92
2,3',4',6-Tetrachlorobiphenyl
17.60
98
292
2,2',3,3'4,4',5,5'6-Nonachlorobiphenyl
26.25
464
89
2,4,4',5-Tetrachlorobiphenyl
17.94
97
Congener
Congener
Table 2: Retention times, AMDIS match factors and SIM ions.

2:116. doi:10.4172/2157-7064.1000116
Page 4 of 8
allowed to soak the sorbent bed for 5 minutes before it was allowed to
pass through. The sample was loaded onto the cartridge, washed with
5 mL of hexane and eluted with 10 mL of hexane. The initial steps of
sample pre-treatment were followed, as in the supelclean sulfoxide
cartridges, i.e. spiking of oil sample and acid treatment.
Results and Discussion

A mixture of 39 PCB congeners i.e. mix C-CS-01, 10 g/mL which
was used through out this work was identified using AMDIS version
3.2 developed by NIST. Broadly and as a rule of the thumb, a simple
match factor between 70 and 80% indicated a good match, between 80
and 90% is a very good match and between 90 and 100% is an excellent
match and thus confirms the presence of the analyte of interest. Results
of the deconvolution using a combination of parameters are shown in
Table 2. Table 2 also shows the ions that were selected to be used in all
subsequent SIM experiments. The minimum match factor was that of
2,2,3,4,5,6-hexachlorobiphenyl, at 87%. Almost all the other congeners
had match factors above 90% with the highest being 100%. As seen from
Table 2, there was an unusual elution pattern which was observed. The
general elution pattern as has been observed previously [24] showed an
increase in the retention time with an increase in the number of chlorine
atoms. However, 2,2,3,3,4,5,6-heptachlorobiphenyl with 7 chlorine
atoms eluted earlier than 2,2,4,4,5,5-hexachlorobiphenyl with 6
chlorine atoms which was confirmed by AMDIS. We speculated that
this was due to the distribution of the chlorine atoms around the two
benzene rings. Even though the hexachlorobiphenyl has less chlorine
atoms, the benzene ring for the heptachlorobiphenyl is hindered by
the chlorine atoms thus preventing interaction with the non polar
stationary phase material and thus results in the quicker elution of the
heptachlorobiphenyl. In contrast, the 2,2,4,4,5,5-hexachlorobiphenyl
congener, the benzene rings are less esoterically hindered by the chlorine
atoms thus increasing interactions with the non polar stationary phase
resulting in longer retention of this congener.
The Optimized conditions for the Supelclean Sulfoxide and Florisil
SPE cartridges were determined as shown in Table 3 below.
Generally, a combination of the above settings gave higher signal
recoveries for all the 39 PCB congeners in the standard mixture.
Recoveries using the two cartridges with optimum extraction

conditions
Figure 1 shows the recoveries obtained. From the results, the florisil
cartridge exhibited selectivity towards early eluting PCB congeners,
while the SupelcleanTM Sulfoxide cartridge showed on average
reasonable recoveries for all PCB congeners. This effect confirms
the non-selectivity of the SupelcleanTM Sulfoxide cartridge whose
adsorption properties are hinged on the interaction of the electrophilic
sulfur atom and the electron cloud arising from the aromatic rings of
the PCB congeners, implying the chlorine atoms do not significantly
affect this interaction.
Comparison of full and SIM scan modes

Since the SupelcleanTM Sulfoxide cartridge showed the best
extraction characteristics of the two cartridges, experiments were
designed to compare full and SIM scan modes performance of the
mass spectrometer. Validation and real sample analysis experiments
were also done using the SupelcleanTM Sulfoxide cartridge because
of this observation. Initially, the peak areas obtained by running a
sample extract which was spiked with 0.20 ppm of the 39 congener
mix standard and initially screened and found to contain no PCBs, in
Parameter
Supelclean Sulfoxide SPE

Cartridges
Florisil SPE Cartridges
Eluting solvent
1:1 (v/v) n-hexane:acetone
1:1 (v/v) n-hexane:acetone
Flow rate
2 mL/min
1 mL/min
Load volume
3 mL
3 mL
Table 3: Optimum conditions used in the SPE extraction of PCB congeners.
full and SIM scan modes. The results of this comparison are shown in
Figure 2 below.
Results from these experiments showed significant signal loss
when SIM is employed as the scan mode. This phenomenon has been
observed before [25] since isolation of one or a set of ions implies signal
loss compared to full scan where all ions can be used for quantification
purposes. However, when method detection limits (MDLs) were
compared using full scan and SIM, the opposite was observed as seen
in Figure 4. This was due to the enhanced selectivity that SIM affords
and thus ultimately compensates the loss in sensitivity due to signal
loss as seen in Figure 3 which is a contrast of peak areas obtained in the
full and SIM scan modes.
Figure 4 shows a comparison of the MDLs, which were determined
by extracting a spiked transformer oil sample, with 0.2 ppm of the
39-congener mix standard and which was initially screened and found
to contain no PCBs. The spiked sample extracts were analyzed in full
and SIM scan modes using the SupelcleanTM Sulfoxide SPE cartridge.
The runs for these extracts were repeated 7 times in each scan
mode and the run for the pure 0.2 ppm standard repeated 3 times.
The peak areas and the standard deviation for seven injections were
computed, and the corresponding peak areas for the extracts converted
to their corresponding concentrations using the peak areas for the
0.2 ppm of the 39 congener mix standard. The MDLs were calculated
by multiplying the standard deviation, s, and the student t-value at 6
degrees of freedom and 95% confidence limit as shown in Equation 1
below:
MDL = ts
(1)
Figure 5a is a full scan total ion chromatogram (TIC) of the 39 PCB

congeners while Figure 5b is a SIM ion chromatogram of the same. The
two chromatograms show the selectivity that SIM affords in contrast
to the earlier.
Method Validation
Results obtained for the instrument detection limits (IDLs), limits
of quantitation (LOQs), MDLs and r2 values, are shown in Table 3.
IDLs were obtained in a similar manner as the MDLs except that pure
standards were used instead of sample extracts for the seven injections
that afforded the required standard deviations used in Equation 1.
In contrast to MDLs, IDLs were lower followed by LOQs. This was
expected because there is only the noise arising from the instrument
when IDLs are evaluated as opposed to two noise components i.e. from
the instrument and the matrix in the evaluation of MDLs. The IDLs
ranged from 0.004 to 0.08 ppm. The MDLs ranged from 0.02 to 0.13
ppm. The LOQs ranged from 0.01 to 0.3 ppm.
The linearities of the calibration curves r2 values were determined
are shown in Table 3. r2 values ranged from 0.9624 to 0.9999. A selected
congener peak was used as an internal standard to quantify all the
other 38 congeners. At the end of this, another congener was selected
and its peak used to quantify the first congener, which was used as an
internal standard for the other 38. The percentage recoveries for the
2-Chlorobiphenyl
3-Chlorobiphenyl
4-Chlorobiphenyl
2,2-Dichlorobiphenyl
2,2,3-Trichlorobiphenyl
2,2,3,5-Tetrachlorobiphenyl
2,2,3,3,4-Pentachlorobiphenyl
2,3,34,6-Pentachlorobiphenyl
2,2,3,4,4,5-Hexachlorobiphenyl
2,2,3,3,4,5,6-Heptachlorobiphenyl
2,2,3,3,4,5,6-Hexachlorobiphenyl
2,2,3,34,5,6-Heptachlorobiphenyl
2,2,3,3,4,45,5-Octachlorobiphenyl
2,2,3,3,4,5,5,6-Octachlorobiphenyl
2,2,3,4,4,5,5,6-Octachlorobiphenyl
2,2,3,34,4,5,56-Nonachlorobiphenyl
Peak Area
2-Chlorobiphenyl
3-Chlorobiphenyl
4-Chlorobiphenyl
2,26-Trichlorobiphenyl
2,3,34,6-Pentachlorobiphenyl
2,2,3,3,4,5,6-Hexachlorobiphenyl
2,2,3,34,5,6-Heptachlorobiphenyl
2,2,3,3,4,5,5,62,2,3,4,4,5,5,62,2,3,34,4,5,56-
%Recovery
2:116. doi:10.4172/2157-7064.1000116
Page 5 of 8
120.00
100.00
80.00
60.00
10.00
20.00
0.00
congeners using the SpelcleanTM Supelco cartridges and SIM are also
shown in Table 4. From the results obtained, the percent recoveries for
the congeners ranged from 44.530.31 to 111.934.75% uncorrected
for bias. Three congeners were recovered at a higher than normal level

FLORISIL
SULFOXIDE
Congener
Figure 2: Comparison of recoveries of PCB congeners obtained by the Waters Florisil and the Supelclean Sulfoxide solid phase cartridges using their optimized
conditions.
80000000
70000000
60000000
50000000
40000000
30000000
20000000
10000000
SIM
0
FULL SCAN
Congener
Figure 3: Comparison of peak areas obtained using the full and SIM scan modes.
of greater than 100%. Analysis of the samples, however, showed that no

PCB congeners were present and as such the greater than 100% recovery
could have been due to instrumental noise resulting in fluctuations in
the peak areas of the analyte peaks in the spiked sample.
2:116. doi:10.4172/2157-7064.1000116
Page 6 of 8
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, -T ch b ny
2,3 4,4 etra loro iphe l
-T c b n
2,3 ,4,5 etra hloro iphe yl
,4 -Te chl bip ny
2 ,6 tra or h l
2,2 ,4,4 -Tet chlo obip enyl
,5 ra r h
2,2 ,3,3 -Tet chlo obip enyl
,3 ,4- rac rob hen
2,2 ,4, Pen hlo ip yl
5 t r he
2, ,4,4 -Pe achl obip nyl
2,2 3,3, ,5-P ntac orob hen
4 e h i y
2,2 ,3,4 ,6-P ntac lorob phen l
,3 ,4, en hlo iph yl
2, ,4 5- tac rob en
2,2 2,3 ,5,5 Hex hlor iph yl
,3 ,4, -H ach obi eny
p
5 e
2,2 ,3,4 ,6-H xac lorob hen l
2,2 ,4 ,5, ex hlo iph yl
,3 ,4 6-H ach rob en
2,2 ,3, ,5,5 exa lor iph yl
c o e
4
2,2 ,3,3 ,5,6 -Hex hlor biph nyl
,3 4, -H ach ob eny
2,2 ,3 5,6 ep lo iph l
, ,5 - tac ro e
2,2 3,4,4 ,6,6 Hep hlor biph nyl
,3 ,5 -H tach ob eny
,4 ,5 ept lo iph l
,5, -H ac rob en
5, ep hlo ip yl
6-H tac ro hen
ep hlo biph yl
tac ro en
hlo bip yl
rob hen
iph yl
en
yl
l l l l l
ny n y n y n y n y
he h e h e h e h e
ib p bip bip bip bip
o o o o o
or o r o r o r o r
hl c h l a c h l c h l c h l
c
a a
a
a
ct c t c t c t o n
-O , 6 - O 6 - O , 6 - O 6 - N
5
,
5, 5 5 , 5 , 5
,4 4 , 4 , 5 , , 5 , 5
,4 , 3 , 3 , 4 , 4 , 4 4 , 4
3
,
3 ,
,3 , 2 , , 3 2 , 3 , 3 , 3
2,2 2 2 , 2 2 , , 2
2
MDLFULL
SCAN
MDLSIM
Congcncr
Figure 4: Comparison of MDLs obtained using the full and SIM scan modes.
Application to real world samples

All six samples that were analyzed using the method described
here were found to contain no PCBs as shown in Table 4. These results
were compared with the ones obtained using the Dexsil L2000 DX,
version 1.27 PCB/chloride analyzer. The Dexsil L2000 PCB/chloride
analyzer extracts PCBs from transformer oil and dissociates them
using a sodium reagent thus freeing chloride ions. The instrument then
uses a chloride ion selective electrode (ISE), employing the 1242 oil
method, which is embedded in the instrument [26]. Results that were
originally obtained by this instrument are also shown in Table 4. The
results showed that the transformer oil samples contained PCBs at the
levels indicated. Perhaps the reason for this is that, the DX analyser
responds to the presence of chloride ions in solution. This implies that
any chloride in a sample regardless of its source could be detected to
Time-->
TIC: Saidy 198.D\datasim.ms
70000
65000
60000
55000
50000
45000
40000
35000
30000
25000
20000
15000
10000
5000
Time-->
6.00 8.0010.0012.0014.0016.0018.0020.0022.0024.0026.0028.00
Figure 5b: SIM ion chromatogram of the 39 PCB congeners.
Abundance
700000
650000
600000
550000
500000
450000
400000
350000
300000
250000
200000
150000
100000
50000
Abundance
TIC: Saidy 198.D\data.ms
give a false positive. The method developed in this study gave 0.0189
ppm as the lowest MDL, an inference that no PCBs were present in the
samples as stipulated by the SC can be made and thus the results given
by the DX analyzer were false positives.
Conclusions
6.00 8.0010.0012.0014.0016.0018.0020.0022.0024.0026.0028.00
Figure 5a: Full scan total ion chromatogram (TIC) of the 39 PCB congeners.

A rapid, selective and sensitive procedure for the determination

of PCBs in transformer oil has been developed, validated and tested
on real samples. The only disadvantage of the method would relate to
the cost, service and maintenance of the instrumentation involved. The
procedure proved to be robust and can be adopted for use in the SADC
region where screening for POPs such as PCBs is mandatory due to
the fact most of the countries that make up the region are party to the
SC. The results obtained from real samples showed that the test kits for
preliminary investigations to aid establishment of baseline inventories
in the formulation of NIPs for SADC State Parties to the SC could have
2:116. doi:10.4172/2157-7064.1000116
Page 7 of 8
IDL = yb + 3sb (ppm
w/v)
PCB CONGENER
LOQ = yb + 10sb (ppm w/v)
r2
Regression Equation (y= a +bx)
MDL = ts
(ppm w/v)
Percent
Recoveries
2-Chlorobiphenyl
0.0589
0.1962
0.9811
y = 0.590x + 0.875
0.1261
44.530.31
3-Chlorobiphenyl
0.0589
0.1962
0.9811
y = 0.590x + 0.876
0.1142
48.200.09
4-Chlorobiphenyl
0.0360
0.1200
0.9929
y = 0.49x + 0.905
0.0610
46.070.05
0.0041
0.0138
0.9999
y = 1.13x + 0.405
0.0820
79.000.82
0.0408
0.1360
0.9908
y = 0.832x + 1.185
0.1044
54.670.30
2,4'-Dichorobiphenyl
0.0679
0.2264
0.9750
y = 0.0467x + 1.25
0.0621
69.280.52
76.491.60
2,5 -Dichlorobiphenyl
0.0309
0.1031
0.9947
y = 0.175x + 1.187
0.0283
0.0213
0.0710
0.9975
y = 0.0784x + 1.741
0.0536
81.950.79
0.0085
0.0284
0.9996
y = 0.105x + 1.538
0.0931
103.531.63
0.0478
0.1593
0.9875
y = 0.281x + 1.707
0.0837
81.601.06
2,3,4'- Trichlorobiphenyl
0.0110
0.0368
0.9993
y = 0.344x + 1.249
0.0779
68.260.92
71.830.13
0.0808
0.2692
0.9650
y = 0.595x + 1.191
0.0966
2,4,4'-Trichlorobiphenyl
0.0503
0.1678
0.9861
y = 16.37x - 0.422
0.1050
80.030.63
0.0762
0.2539
0.9688
y = 3.686x + 0.690
0.1006
80.330.47
0.0339
0.1132
0.9936
y = 3.437x + 0.776
0.0740
78.050.83
2,3,3',4'-Tetrachlorobiphenyl
0.0839
0.2796
0.9624
y = 0.989x + 1.05
0.0377
81.882.28
0.0389
0.1298
0.9917
Y= 0.789x + 1.12
0.0966
62.300.21
2,3',4,5-Tetrachlorobiphenyl
0.0309
0.1030
0.9947
y= 0.805x + 1.09
0.0383
54.270.52
2,3',4',6-Tetrachlorobiphenyl
0.0544
0.1812
0.9839
y= 2.88x + 0.61
0.0587
76.980.52
2,4,4',5-Tetrachlorobiphenyl
0.0503
0.1676
0.9862
y = 9.75x + 0.106
0.0714
82.110.69
0.0537
0.1789
0.9843
y = 3.019x + 1.12
0.0701
71.691.20
2,2',3,4,5'-Pentachlorobiphenyl
0.0096
0.0321
0.9995
y = 2.42x + 1.28
0.0846
103.610.43
0.0198
0.0660
0.9978
y = 0.657x + 1.19
0.0301
80.470.80
2,3,3',4',6-Pentachlorobiphenyl
0.0275
0.0916
0.9958
y = 1.02x + 1.52
0.0722
111.934.75
2,2',3,4,4',5'-Hexachlorobiphenyl
0.0225
0.0750
0.9972
y = 1.23x + 1.96
0.0587
57.312.04
0.0317
0.1055
0.9945
y = 0.982x + 1.74
0.0927
69.642.84
2,2',3,4',5,6-Hexachlorobiphenyl
0.0361
0.1202
0.9928
y = 0.803x + 1.51
0.0583
72.162.16
2,2',3,3',4,5,6-Heptachlorobiphenyl
0.0239
0.0795
0.9968
y = 1.65x + 1.70
0.0252
68.441.79
0.0038
0.0127
0.9999
y = 1.25x + 1.76
0.0189
84.680.29
0.0160
0.0533
0.9986
y = 0.878x + 1.82
0.0490
66.630.98
2,2',3,3'4',5,6-Heptachlorobiphenyl
0.0324
0.1079
0.9942
y = 2.59x + 2.59
0.0697
76.321.12
0.0077
0.0255
0.9997
y = 4.31x + 0.957
0.0386
68.030.67
2,2',3,4,4',5,5'-Heptachlorobiphenyl
0.0419
0.1397
0.9903
y = 1.93x + 2.18
0.0581
69.691.06
2,2',3,4',5,5',6-Heptachlorobiphenyl
0.0270
0.0899
0.9960
y = 0.612x + 2.084
0.0579
53.521.69
2,2',3,3',4,4'5,5'-Octachlorobiphenyl
0.0288
0.0961
0.9954
y = 1.13x + 2.03
0.0435
68.942.11
2,2',3,3',4,4'5,6-Octachlorobiphenyl
0.0192
0.0641
0.9979
y = 2.64x + 1.44
0.0349
54.710.80
2,2',3,3',4,5,5',6'-Octachlorobiphenyl
0.0673
0.2244
0.9754
y = 3.78x + 0.0997
0.0508
76.383.43
2,2',3,4,4',5,5',6-Octachlorobiphenyl
0.0162
0.0539
0.9985
y = 1.93x + 0.231
0.0530
72.47 0.80
2,2',3,3'4,4',5,5'6-Nonachlorobiphenyl
0.0098
0.0326
0.9995
y = 2.51x + 0.0705
0.0420
54.083.43
Table 4: IDLs, LOQs, MDLs,r 2 values, regression equations and percent recoveries using the SupelcleanTM Sulfoxide cartridge for the 39 PCB congeners.
been unreliable. This places the need to re-screen all suspected PCB
containing equipment in the SADC region using reliable methods like
the one proposed here and compare them to baseline inventories that
were established to aid in developing National Implementation Plans
(NIPs).
GC-MS results
(PCB congeners)
Dexsil L2000 DX analyzer (ppm)
SSKA 004
ND
116
SSKA 009
ND
329
SSKA 024
ND
456
SSKB 002
ND
109
Glen valley 006
ND
192
Mmamashia 003
ND
365
Sample
ND= Not Detected.

See sampling sites coordinates in Table 1
Table 5: Results of six real transformer oil samples.

Acknowledgements
The authors would like to acknowledge the University of Botswana Vice
Chancellor, Deputy Vice Chancellor (Finance), Faculty of Science Dean and his
Deputy, and all those that facilitated the purchase of the Agilent 5975C GC-MS
system which was used in this work and housed in the Mass Spectrometry Unit.
References
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2. Martinez HA, Rodriquez GC, Castillo DH (2005) Determination of PCBs in
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Chromatography

Motladiile et al., J Chromatograph Separat Techniq 2011, 2:4

Development and Validation of a Gas Chromatography-Mass

Keywords: PCBs; GC-MS; Botswana; Stockholm convention; SADC;

Investigations in many parts of the world have revealed widespread

to the increased awareness that persistent organic pollutants (POPs)

Due to their physical and chemical properties, PCBs are highly

*Corresponding author: Saidy Motladiile, Department of Waste management

National implementation plan

ReceivedDecember 02, 2011; Accepted January 03, 2012; Published January

J Chromatograph Separat Techniq

Citation: Motladiile S, Kwaambwa HM, Sichilongo K (2011) Development

Volume 2 Issue 4 1000116

acquired from Biotage, Charlotte, NC, USA. This is a vacuum manifold

Sample collection and storage

Volume 2 Issue 4 1000116

Glen valley 006

*n/a- not available

Supelclean Sulfoxide SPE cartridges: The SPE normal procedure

Table 2: Retention times, AMDIS match factors and SIM ions.

J Chromatograph Separat Techniq

Volume 2 Issue 4 1000116

Results and Discussion

Recoveries using the two cartridges with optimum extraction

Comparison of full and SIM scan modes

Supelclean Sulfoxide SPE

Florisil SPE Cartridges

1:1 (v/v) n-hexane:acetone

1:1 (v/v) n-hexane:acetone

Table 3: Optimum conditions used in the SPE extraction of PCB congeners.

Figure 5a is a full scan total ion chromatogram (TIC) of the 39 PCB

Volume 2 Issue 4 1000116

J Chromatograph Separat Techniq

of greater than 100%. Analysis of the samples, however, showed that no

Volume 2 Issue 4 1000116

Application to real world samples

TIC: Saidy 198.D\datasim.ms

TIC: Saidy 198.D\data.ms

J Chromatograph Separat Techniq

A rapid, selective and sensitive procedure for the determination

Volume 2 Issue 4 1000116

LOQ = yb + 10sb (ppm w/v)

Regression Equation (y= a +bx)

Dexsil L2000 DX analyzer (ppm)

Glen valley 006

ND= Not Detected.

J Chromatograph Separat Techniq

Volume 2 Issue 4 1000116

DDTs and methyl sulphone metabolites in various tissues of harbour porpoises

16. Karlson K, Ishaq R, Becker G, Berggren P, Broman D, et al. (2000) PCBs,

Submit your next manuscript and get advantages of OMICS

User friendly/feasible website-translation of your paper to 50 worlds leading languages

200 Open Access Journals

Submit your manuscript at: http://www.editorialmanager.com/biochem

J Chromatograph Separat Techniq

Volume 2 Issue 4 1000116

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