"Science Stays True Here"

Biological and Chemical Research, Volume 2015, 111-122 | Science Signpost Publishing

Determination of Total Hydrocarbons and Alkyl PAHs in

Sediments from Apsheron Peninsula in Caspian Sea
Navai A. IBADOV and Bahruz A. SULEYMANOV
Institute of Radiation Problems of Azerbaijan NAS, 121 H.Javid aven., AZ1143,Baku, Azerbaijan.

Received: January 08, 2014 / Accepted: February 06, 2015 / Published: March 25, 2015
Abstract: Investigation of total petroleum hydrocarbons (THC) and polycyclic aromatic hydrocarbons (PAHs) in sea sediment
samples that were collected from the areas of the Caspian Sea. Samples are extracted with methylene chloride and methanol, after the
extracts are cleaned on silica-gel columns and then injected into GC/FID for determination of THC and GC/MS operating in the
selected-ion-monitoring (SIM) mode for determination of the PAHs and their alkylated.
Keywords: THC, PAHs, GC/MS, SIM, RRF, Sediment

1. Introduction
Most methods for measuring the semivolatile compounds of petroleum in environmental samples first extract
them with an organic solvent, fractionate with chromatography, and then analyze the fractions by infrared (IR)
spectroscopy, ultraviolet fluorescence (UVF) spectroscopy, or gas chromatography (GC) [1]. Each of these
methods targets a particular characteristic of the extracted material and gives an operationally-defined
concentration of total hydrocarbons (THC). Although IR and UVF provide some useful information and a
measure of THC values, they lack the specificity and sensitivity of GC methods.
Polycyclic aromatic hydrocarbons (PAHs) are recognized as potent carcinogens and numerous studies have
shown that they are ubiquitous contaminants in a wide variety of matrices such as air, food, fly ash, soil,
sediments, water, crude oil and petrochemicals [25].
There are a wide variety of solvent extraction techniques commonly used for extracting hydrocarbons from
soils and sediments. Traditional extraction procedures include Soxhlet [69] ultrasonication [7,10,11], mechanical
shaking [12,13]. Modern techniques include supercritical fluid extraction, pressur-ized liquid extraction,
accelerated solvent extraction, microwave assisted extraction and focused microwave assisted. Each technique has
its own merits and the choice of extraction depends on several factors including capital cost, operating cost,
simplicity of operation, amount of organic solvent required, sample throughput and the availability of a
standardized method.
In this work used ultrasonic procedure for extracting hydrocarbons and PAHs in sea sediments.
Polynuclear aromatic hydrocarbons (PAHs) are important environmental pollutants because many of them are
known or suspect carcinogens. In environmental studies, the extent of PAH contamination as a group is often
Corresponding author:
Navai A. IBADOV, Institute of Radiation Problems of Azerbaijan NAS, 121 H.Javid aven., AZ1143,Baku, Azerbaijan. E-mail:
navai@azecolab.com.

112

The Impact of Oil Contamination on Soil Ecosystem

quantified by the concentrations of 16 of the representative PAH species, which are included in the list of priority
pollutants as defined by USEPA. Many analytical techniques have been developed in the past years for the
determination of these 16 PAH species, and among them two of the most widely practiced ones are
HPLC-Fluorescence [14] and GC/MS [15]. HPLC with fluorescence detection (HPLC-FLD) cannot be used for
the detailed analysis of individual alkylated PAHs. Gas chromatography with mass spectrometry (GC-MS) is
presently the preferred analytical technique for the analysis of both parent and alkylated PAHs [17]. As is often
the case in environmental analysis, effective sample preparation and cleanup holds the key of success for the
analysis of trace PAHs.
Aromatic ring structures in petroleum products range from one- to five-ring combinations. Two or more fiveor
six-member carbon rings are fused together to form polyaromatic hydrocarbons (PAHs). These petroleum PAHs
have abundant alkyl group substitution on their ring structures. The alkyl groups generally have one to four
saturated carbon atoms, and thus can produce many different structural isomers and homologs for each aromatic
hydrocarbon family. The most abundant aromatic hydrocarbon families have two or three fused rings with one to
four carbon atom alkyl group substitutions (denoted C1-, C2-, C3-, and C4- by GC/MS/SIM expanded scans). It is
important to point out that crude oils contain primarily the alkyl homologs of aromatic compounds and relatively
small quantities of the unsubstituted "parent" aromatic structures. C1-, C2-, C3-, or C4- followed by a PAH name
(for example, C1-naphthalene) is a naming convention for reporting the total of all detected C1-, C2-, C3-, or
C4-alkyl homologs of the noted PAH. For example, C1-naphthalene reported concentrations represent the total
concentration of all C1 naphthalenes. C1-compounds differ from C2-compounds in that there is one rather than
two carbon groups attached. Groups of alkyl homologs are often analyzed by a GC/MS/SIM expanded scan for
polyaromatic hydrocarbons (PAHs) and alkyl PAHs.
A gas chromatograph/mass spectrometer (GC/MS) in selected ion mode (SIM) coupled to a capillary column is
used to resolve, detect and quantitate polycyclic aromatic hydrocarbons (PAH) in solids at parts per billion levels.
Samples are injected into a temperature-programmed GC/MS, operated in splitless mode. The capillary column is
a DB-5MS (30 m x 0.25 mm ID and 0.25 mm film thickness). The mass spectrometer is capable of scanning from
50 to 500 AMU every second or less and uses 70 electron volts energy in electron impact ionization mode. The
data acquisition system continuously acquires and stores all data analyses [15-18].

2. Materials and Methods

Internal and Surrogate Standard Solution
Hexadecane-d34 (10 g, 5-alpha-Androstane, and squalane (Sigma-Aldrich) as internal and surrogate standards
for hydrocarbon analysis by GC-FID. Added 10 g of each hexadecane-d34 and squalane and 5 g of
5-alpha-Androstane.
The standard solutions is made from aliquots of mixture (Z-014J-PAK; M-525SS-PAK; M-8310-SS-PAK,
AccuStandard, Inc., USA) or pure compounds and diluted with dichloromethane to a final concentration of 0.25
ug/mL. The standards solution includes naphtalene-d8, acenaphthene-d10, phenantrene-d10, pyrene d10,
chrysene-d12 and perylene-d12. The internal standard compounds are resolved from, but elute in close proximity
to the analytes of interest. The internal standard solution is added to all samples and quality control samples just
prior to instrument analysis. Internal standards are used to calculate relative response factors and specific analyte
concentrations based on retention time.

The Impact of Oil Contamination on Soil Ecosystem

113

Matrix Spiking Solution

For the THC spike solution used C7-C40 Saturated Alkanes (Sigma Aldrich, Cat#49452-U) Mineral Oil mix
A&B for ISO 9399-2 Sigma Aldrich, Cat#18602.
Certified solutions containing 2 to 6-ring PAH compounds are purchased from commercial vendors
(Benz(a)pyrene,
Chrysene,
1-Methylnaphthalene,
2-Methylnaphthalene,
Phenanthrenes,
Pyrene;
Cat.M-610-MS-PAK, AccuStandard, Inc., USA) and diluted with dichloromethane to prepare the matrix spiking
solution. The matrix spiking solution is diluted and is added to all matrix spike samples.

Calibration Solution
For GC/FID calibration mix used C7-C40 Saturated Alkanes (Sigma Aldrich, Cat#49452-U). Calibrations
solutions (16EPA PAHs and their 34 individual Alkyl PAHs components, AccuStandard, Inc. or Sigma-Aldrich))
are prepared at 7 concentrations ranging from approximately 0.01 to 1 g/mL, by diluting commercially available
certified solutions containing analytes of interest.

Identifications and Quantitative Determination of THC

Identification and determination of THC used measurement of area records for C10-C20 and C21-C40 and
calculations of base of hexadecane-d34 and squalane internal standards. RRF(C10-C20) =1.13 and RRF(C21-C40) =1.02.

Identifications and Quantitative Determination of PAHs By GC/MS-SIM

Mass Spectrometer Tuning
Prior to calibration, the MS is autotuned to perfluorotributylamine (PFTBA) using criteria established by the
instrument manufacturer.

Initial Calibration
Prepare calibration standards at a minimum of seven concentration levels for each parameter (Naphthalene,
2-Methylnaphthalene, 1,3-Dimethylnaphthalene, Acenaphthylene, Acenaphthene, Fluorene, Dibenzothiophene,
Phenanthrene, Anthracene, 4-Methyldibenzothiophene, 1-Methylphenanthrene, 3,6-Dimethylphenanthrene,
Fluoranthene,
Pyrene,
2-Methylfluoranthene,
1-Methylpyrene,
Benz[a]Anthracene,
Chrysene,
9-Methylbenz[a]Anthracene, 6,8-Dimethylbenz[a]Anthracene, Benzo[b]fluoranthene, Benzo[k]fluoranthene,
Benzo[a]Pyrene, Perylene, 9-Methylbenzo[a]pyrene, 7,10-Dimethylbenzo[a]pyrene, Indeno(1,2,3-cd)pyrene,
Benz[g,h,i]perylene, Dibenz[a,h]Anthracene) of interest by adding volumes of one or more stock standards to a
volumetric flask and diluting to volume with dichloromethane. The following calibration levels are used: 2, 5, 10,
25, 50, 100 and 200 mg/L for the individual hydrocarbons and 0.010, 0.020, 0.050, 0.100, 0.200, 0.500 and 1.000
g/L for the PAH components. A 7-point relative response factor (RRF) calibration curve is established for
analytes of interest prior to the analysis of samples and quality control samples.
A RRF is determined, for each analyte, for each calibration level using the following equation 1:

RRF =
Where:

Astd Amount(IS)
Ais Amount(S)

(1)

114

The Impact of Oil Contamination on Soil Ecosystem

Astd peak area of the standard;

Amount(S) amount of standard, ng;
Amount(IS) amount of internal standard added to the sample and standard solutions, ng:
Ais peak area of the internal standard.

The response factors determined for each calibration level are averaged to produce a mean relative response
factor (RRF) for each analyte. The percent relative standard deviation (%RSD) for the 7 response factors must be
less than or equal to 25%, for each analyte.

Continuing Calibration Check

A mid-level calibration standard is analyzed at the beginning and end of each analytical set samples. The daily
relative response factor for each compound is compared to the mean relative response factor from the initial
calibration curve and the average relative percent difference (RPD) of all analytes must be less than 25%. If the
calibration check does not meet this criterion then the initial fivepoint calibration is repeated.

GC/FID ANALYSIS
The hydrocarbons are analyzed on a gas chromatograph with a flame ionization detector (GC-FID), and the
THCs are usually estimated by integrating the areas of the resolved and unresolved components [19].
Chromatographic separation of THC was accomplished on a CP-Sil 5CB (100% dimethylpolysiloxane) capillary
column (15 m0.25 mm I.D., 0.25 mm film thickness). Nitrogen was the carrier gas and a pressure 15 psi
(constant) was used for column elution. Sample injection was carried out in the splite/splitless mode with an
injection volume of 1 L. Split ON at 0.75min, Split Ratio-20. Autosampler CP8410 (Varian). The GC oven
temperature was programmed first from 40C (hold 2 min), 40C to 320C at a rate of 10C/min (hold 2.0 min).
Fid temperature were 340C (Make-up- 30 ml/min Hydrogen-30 ml/min, Air 300 ml/min ). The temperatures of
the injection port were set at 300C. Galaxie software acquisition. Capable of continuous acquisition and storage
of all data during analysis.

GC/MS-SIM ANALYSIS
The initial calibration of the GC/MS must meet the previously described criteria prior to sample analysis.
Samples are analyzed in analytical sets that consist of standards, samples and quality control samples. Quality
control samples are method blanks, laboratory duplicates, blank spikes, matrix spikes and standard reference
materials. The type and number of quality control samples depend upon client requests and material availability.
Thermo-Electron GC/MS Trace DSQ System equipped with a CombiPal Auto Sampler was used for analysis.
Chromatographic separation of 2-6 ring PAHs was accomplished on a DB-5MS capillary column (30 m0.32 mm
I.D., 0.25 mm film thickness). Helium was the carrier gas and a flow-rate of 1.2 mL/min was used for column
elution. Sample injection was carried out in the splitless mode with an injection volume of 1 L. The GC oven
temperature was programmed first from 40C (hold 1 min), 40C to 120C at a rate of 15C/min, then to 256C at
a rate of 6/min (hold 5.0 min), and then to 300C at a rate of 6/min finally held constant for 5 min. The
temperatures of the injection port and the interface to the MS system were set at 300C and 300C, respectively.
Peak quantification was carried out in Selected Ion Monitoring (SIM) mode. MS Ion source temperature at 250C.
Software - ThermoElectron-Finnigan Technologies Xcalibur 1.4SR1, capable of continuous acquisition and

The Impact of Oil Contamination on Soil Ecosystem

115

storage of all data during analysis. For identifications of components used 4 segments (in the each segments
present 12 ions), for each segments scan time equal to 1.39s and dwell time 0.1 sec per ion.

Analyte Identification for Alkyl PAHs

The extracted ion current profiles of the primary m/z and the confirmatory ion for each analyte must meet the
following criteria:
The characteristic masses of each analyte of interest must be in the same scan or within one scan of each other.
The retention time must fall within 5 seconds of the retention time of the authentic compound or alkyl
homologue grouping determined by the analysis of the daily calibration check.
The alkylated PAH homologue groupings (e.g. C4-naphthalene) appear as a group of isomers. The pattern of
each group and the retention time window for the group is established by the analysis of a reference oil standard.
Each group of alkylated homologues is integrated in its entirety and the total area response is used to determine
the concentration of the entire group.
The relative peak areas of the primary mass ion, compared to the confirmation or secondary mass ion, must fall
within 30 percent of the relative intensities of these masses in a reference mass spectrum (Table 1). The
reference mass spectrum is obtained from the continuing calibration solution or the reference oil standard for the
parent compounds and alkylated homologues, respectively. In some instances, a compound that does not meet
secondary ion confirmation criteria may still be determined to be present in a sample after close inspection of the
data by a qualified mass spectrometrist.
Table 1. Target Analyte Parameters.

Analyte

Quantitation
Ion, m/z

Reference to
Internal Standard

RRF

and Surrogate

Naphthalene

128

Naphthalene-d8

1.22

C1-Naphthalenes

142

Acenaphthene-d10

1.07

C2-Naphthalenes

156

Acenaphthene-d10

1.04

C3-Naphthalenes

170

Acenaphthene-d10

1.04

C4-Naphthalenes

184

Acenaphthene-d10

1.04

Acenaphthylene

152

Acenaphthene-d10

1.50

Acenaphthene

153

Acenaphthene-d10

1.25

Fluorene

166

Acenaphthene-d10

1.31

Dibenzothiophene

184

Phenanthrene-d10

1.18

C1-Dibenzothiophenes

198

Phenanthrene-d10

0.86

C2-Dibenzothiophenes

212

Phenanthrene-d10

0.86

C3-Dibenzothiophenes

226

Phenanthrene-d10

0.86

Phenanthrene

178

Phenanthrene-d10

1.21

Anthracene

178

Phenanthrene-d10

1.01

C1-178 PAHs

192

Phenanthrene-d10

1.07

C2-178 PAHs

206

Phenanthrene-d10

0.95

C3-178 PAHs

220

Phenanthrene-d10

0.95

The Impact of Oil Contamination on Soil Ecosystem

116

Fluoranthene

202

Pyrene-d10

1.38

Pyrene

202

Pyrene-d10

1.31

C1-202 PAHs

216

Pyrene-d10

1.14

C2-202 PAHs

230

Pyrene-d10

1.14

C3-202 PAHs

244

Pyrene-d10

1.14

Benz[a]anthracene

228

Chrysene-d12

0.86

Chrysene

228

Chrysene-d12

1.19

C1-228 PAHs

242

Chrysene-d12

0.68

C2-228 PAHs

256

Chrysene-d12

0.53

Benzo[b]fluoranthene

252

Perylene-d12

1.78

Benzo[k]fluoranthene

252

Perylene-d12

1.88

Benzo[a]Pyrene

252

Perylene-d12

1.13

Perylene

252

Perylene-d12

1.08

C1-252 PAHs

266

Perylene-d12

0.51

C2-252 PAHs

280

Perylene-d12

0.39

Indeno(1,2,3-cd)pyrene

276

Perylene-d12

1.00

Benz[g,h,i]perylene

276

Perylene-d12

1.32

C1-276 PAHs

290

Perylene-d12

1.00

C2-276 PAHs

304

Perylene-d12

1.00

Dibenz[a,h]Anthracene

278

Perylene-d12

1.07

Data not meeting the criteria established in this section are appropriately qualified or re-analyzed.

Quantitation Calculations
Sample analyte concentrations are calculated based on the concentration and response of the internal standard
compounds. The equations 1 used to calculate the RRF of each analyte relative to the concentration and area of
the internal standard in the initial calibration. Response factors for same alkyl homologues are presumed equal to
the average response factor of the respective unsubstituted (parent) compound.
The concentration of TPH or PAHs components in the original wet sample, Cw is calculated from the following
equation 2:

Cw =

As Amount(IS)
Ais RF Msamp

(2)

Where:
Cw - Concentration of compound of interest in the wet sample, ng/g for PAHs compounds;
As peak area of the compound of interest;
Ais peak area of the corresponding internal standard;
Amount(IS) the amount of internal standard added to the sample, blank, calibration satandard solution and QC samples
in 1 ml of extract,ng;
RF the response factor; and
Msamp mass of wet sample taken for extraction, g.

The Impact of Oil Contamination on Soil Ecosystem

117

To convert the result to a dry matter basis, the following equation is used:

Cd = Cw

100
DM

(3)

Where:
Cd concentration of compound of interest in the dry sample (ng/gdm for PAHs compounds);
Cw - concentration of compound of interest in the wet sample (ng/g for PAHs compounds); and
DM the percent dried weight of the sample.

Quality Control
The initial calibration must pass established criteria before sample analysis can begin. All continuing
calibration checks must pass established criteria for analysis to continue. An acceptable method blank analysis
may not contain more than two target analytes at concentrations three times greater than the MDL. This criterion
does not apply if the analytes detected in the method blank are not detected in the associated samples or if the
sample analyte concentrations are 10 times greater than the blank analyte concentrations. If the method blank
exceeds these criteria then the analytical procedure is not in control. The source of the contamination must be
investigated, and corrective measures taken and documented before further sample analysis occurs.
All samples and quality control samples are spiked with deuterated PAH standards compounds prior to
extraction. The deuterated compounds evaluate sample matrix effects and analytical efficiencies associated with
sample preparation and analysis. The recovery of deuterated surrogate compounds is monitored in each sample
and quality control sample. The laboratory will take corrective action if the average surrogate recovery, with the
exception of perylene-d12, is less than 40% or greater than 140%. The following corrective action will be taken if
the above criteria are not met:

Calibration checks solution. If relative percent difference (RPD) of any analyte within a calibration check
standard varies from the predicted response by more than 25%, a new calibration curve must be prepared
for that analyte -per 10 samples

Laboratory Blank -per 20 samples. The criteria less than MDL (for TPH equal to 1 g/gdm and for PAHs
equal to 0.5 ng/gdm, RSD less than 20% for each components) or less than 30 % of minimal values of
sample.

Quality Control Samples (Certified Reference Material (CRM), Laboratory Fortified Blank (LFB) or
Matrix Spike (MS), depend from lab QC program)- at least per 20 samples. The recovery criteria equal to
40%-140 %.
Sample duplicates (laboratory or field, depend from project)- at least per 20 samples. The % RPD of duplicate
samples must not exceed 50%.

Sampling
Composite sediment samples (including the organic top layer) were taken between 10 july and 20 september
2013 from Apsheron Peninsula in Caspian Sea, South-East of the Baku of the Azerbaijan areas for 50 points (see
fig.1), collected in aluminum pans and transported to the laboratory under cool conditions. Before analysis, the
samples were air homogenized in ambient temperature. No losses of analyts should occur under these conditions
and potential contamination from laboratory air or other samples is considered insignificant.

The Impact of Oil Contamination on Soil Ecosystem

118

Samples are extracted by EPA-Method-3550C (Ultrasonic extraction) and cleaned by EPA-Method-3660B

(Sulfur clean-up) and EPA-Method-3630C (Silica-Gel Cleanup). Concentrate extracts by KudernaDanish
apparatus at 50-55C till 2 ml and transfer extract into volumetric tube. After place the volumetric tube in nitrogen
blow-down apparatus. Evaporate the extract till 1ml and analyzed by GC-FID and by GC/MS (Thermo-Electron,
Finnigan) SIM modes.

Pre-cleaning and conditioning

Checking of lab glassware and materials preparatory to check status each type lab glassware and material
before starting of samples processing according of below chain: Solvent cleaning, Detergent washing,
Acetone treatment, DCM treatment, Drying in oven, Annealing in oven at 400 deg C, Aluminium foiling
and send to storage.

Sodium sulphate is treated as possible before usage and in amount required for batch samples as below:
Bake in a muffle furnace at 400 C for 2 hours. Allow to cool before using. Store sodium sulphate in a large
mouth bottle with cap Glass wool treatment procedure.

Glass wool: Bake in a muffle furnace at 400 C for 2 hours. Store in an oven at 100 C.
Silica Gel treatment procedure: Silica Gel is treated as possible before usage and in amount required for
batch samples as below: 400 deg C, 5 hours. Activating at 200 deg C 1 hours before use and Cooling before
use.

Sample Extraction
Weigh 40g of each sample in Erlenmeyer flasks. The sample was spiked of each of the individual internal
standards and the remaining sediment was added to the Erlenmeyer flask. Add 60 ml DCM and 50 ml methanol
into Erlenmeyer flask and stopper. Now samples are ready for ultrasonic extraction (US) for 30min. After
extraction remove the flask from the US bath and allow the sample to settle. Carefully decant the supernatant
solvent through a glass wool into a 1 liter separating funnel containing 100 ml of distillated water and stopper.
Shake the separating funnel for 1.0 0.1 minute, and allow the layers to separate. Run the lower dichloromethane
layer into a 500 ml round bottomed flask.
Add 50 ml of dichloromethane to the sediment and ultrasonicate for a further 15 minutes. Remove the flask
from the bath and allow the sample to settle. Carefully decant the supernatant solvent through the filter paper into

The Impact of Oil Contamination on Soil Ecosystem

119

the separating funnel containing the residual aqueous methanol. Stopper and shake the separating funnel for 1.0
minute. Allow the layers to separate and combine the dichloromethane extracts by running the lower
dichloromethane layer into the 500 ml round bottomed flask and this process repeats once again and combine the
dichloromethane extracts. Add sufficient (typically 1g) sodium sulfate to the combined dichloromethane extracts
and dry the solvent. When dry, the solvent is filtered through a second (clean) filter paper .Approximately 5-10 ml
of dichloromethane are added to the flask and the flask rinsed. The solvent is then filtered and the extracts
combined.

Filtration and drying through sodium sulfate

Prepare drying column by plugging glass wool into a (d=20mm) column and add Na2SO4 1sm. Elute the
column by 2ml DCM. Place a round bottomed 500ml flask under the column and pour the sample extract. Rinse
the vessel with 20ml DCM by little portions and elute the sodium sulfate to complete the quantitative transfer.
Using Kuderna Danish concentrator concentrated at 60-65C till 2-5 ml. Transfer extract into volumetric tube.
Rinse flask with a little portion of solvent and add the washings to extract to complete the quantitative transfer.
Place the volumetric tube in nitrogen blow-down apparatus. Evaporate the extract till 1ml.

Sulfur Cleanup procedure:

Add approximately 2 g of cleaned copper powder to extract. Vigorously mix the extract and the copper powder
for at least 1 min on the mechanical shaker. Allow the phases to separate. Separate the extract from the copper
into the volumetric tube.

Silica gel Clean-up and extract concentration procedure:

Prepare the chromatographic column by plugging cotton wool into a long bodied Pasteur pipette. Prepare
slurry by adding 20ml DCM to 5gm silica gel. Swirl the contents and add the slurry into the column. Wait
until silica gel will sink. Add 1-2sm sodium sulfate. Open the column tap and discard the solvent. Do not
allow the meniscus of the solvent to fall below the surface of the column.

Transfer the sample extract from volumetric tube to the column. Place a round bottomed 500ml flask
under the column and pour the sample extract. Elute the column 20ml of a 2:1 pentane and DCM mixture.
Rinse the sample extract container with 2-3ml o a mixture and transfer the washings to the column. Elute
the column. Add a further 20ml mixture to the column and continue eluting.

Concentrate extracts by Kuderna Danish apparatus at 50-55C till 2 ml and transfer extract into
volumetric tube.

Rinse flask with a little portion of solvent and add the washings to extract to complete the quantitative
transfer.

After place the volumetric tube in nitrogen blow-down apparatus. Evaporate the extract till 1ml.
The solution is now ready for GC or GC/MS determinations.

Analytical Quality Control

Losses of analytes were compensated by the use of each of their respective deuterated analogues as internal
standards. Limits of quantifications (LOQ) as determined by a signal to noise ratio of 10 in sedimant extract

The Impact of Oil Contamination on Soil Ecosystem

120

chromatograms. Accuracy of the applied method was regularly verified by participation in the WEPALs quarterly
SETOC ring tests.

3. Results and Discussion

Results received for 12 selected tests, are submitted in the table 1.
Table 1

THC

UCM

2-6 PAHs

NPD

(g/g)

(g/g)

UCM

(ng/g)

(ng/g)

ACGR-01

2.4

1.2

52

88

57

64

17

ACGR-A

10.4

8.2

78

111

65

56

29

ACGR-04

3.4

56

158

95

61

40

ACGR-B

84

63

75

454

269

59

113

ACGR-22

221

160

72

524

257

49

132

ACGR-46

1.9

1.3

62

42

26

60

ACGR-62

18

13

75

62

35

57

12

ACGR-W6

6.8

4.8

63

93

62

64

17

ACGR-C

15

12

77

54

33

64

11

ACGR-26

2.9

1.3

45

91

58

64

17

ACGR-33

268

197

74

770

427

55

153

ACGR-D

23

18

77

177

102

58

33

Min

1.9

1.2

45

42

26

49

Max

268

197

78

770

427

64

153

Med

13

10

73

102

64

60

23

Mean

55

40

67

219

124

59

49

Points

%NPD

16 EPA
(ng/g)

There are a few different types of correlations, but the one we will use is the Pearson correlation. A correlation,
r, is a single number that represents the degree of relationship between two measures. The correlation coefficient
is a value such that between from -1 to 1.
A positive correlation indicates a relationship between x and y measures such that as the values of x increases,
the values of y will also increase. A negative correlation indicates the opposite as the values of x increase, the
values of y will decrease. The closer the correlation, r, is to -1 or 1, the stronger x and y are related. The negative
sign does not indicate anything about strength - it just signifies that correlation is negative. If r is close or equal to
0, it means there is a weak or no relationship between the measures.
The relationship between hydrocarbon concentrations was tested by a Pearsons r correlation analysis of
replicate values. Pearson correlation coefficient, also known as Pearson's r, a measure of the strength and direction
of the linear relationship between two variables that is defined as the (sample) covariance of the variables divided
by the product of their (sample) standard deviations.
High correlation coefficients have been given between THC & UCM and between all PAH parameters given in
Table 2:

The Impact of Oil Contamination on Soil Ecosystem

121

Table 2. Pearson correlation coefficient between parameters.

Total 2-6

NPD

Total EPA

THC

UCM

(g/g)

(g/g)

THC (g/g)

1.00

UCM (g/g)

1.00

1.00

% UCM

0.35

0.36

1.00

0.95

0.96

0.33

1.00

NPD (ng/g)

0.93

0.93

0.32

1.00

1.00

% NPD

-0.73

-0.73

-0.56

-0.64

-0.59

1.00

Total EPA 16 (ng/g)

0.94

0.94

0.34

0.99

0.98

-0.69

1.00

Total 2-6 ring PAH

(ng/g)

% UCM

ring PAH
(ng/g)

(ng/g)

% NPD

16PAHs
(ng/g)

4. Conclusion
THC concentration in the 50 points contained from 1.9 mg/kgdw -268 mg/kgdw and UCM present in samples
from 45%-78%. Total 2-6 rings PAHs were present in the sediments observation sites in concentrations from 42
g/kgdw to 770 g/kgdw, 16EPA PAHs concentrations from 9.0 g/kgdw to 153 g /kgdw and NPD from 49 %
to 64% (at 2-6 Total PAHs rings).

References
[1]. NRC (1985) Oil in the Sea: Inputs, Fates, and Effects. National Research Council, National Academy Press, Washington, DC.
[2]. Neff J.M., Polycyclic Aromatic Hydrocarbons in the Aquatic Environment, Applied Science Publishers, London, 1979.
[3]. Bjorseth A. (Ed.), Handbook of Polycyclic Aromatic Hydrocarbons, 1983, p. 727.
[4]. Blumer M.W., Sci. Am. 234 (1976) 34.
[5]. Blumer M., Youngblood W.W., Science 188 (1975) 53.
[6]. Luque de Castro M.D., L.E. Garca-Ayuso, Anal. Chim. Acta. 369 (1998) 1.
[7]. J.O. Grimalt, C. Marfil, Albaiges J., Int. J. Environ. Anal. Chem. 18 (1984) 183.
[8]. Lee H.-B., Dookhran G., Chau A.S.Y., Analyst 112 (1987) 31.
[9]. Wong M.K., Williams J.P.B., Mar. Chem. 9 (1980) 183.
[10]. Luque-Garca J.L., Luque de Castro M.D., Trends Anal. Chem. 22 (2003) 41.
[11]. Krahn M.M., Ylitalo G.M., Joss J., Chan S.-L., Mar. Environ. Res. 31 (1991) 175.
[12]. Berset J.D., Ejem M., Holzer R., Lischer P., Anal. Chim. Acta 383 (1999) 263.
[13]. MacLeod W.D., Brown D.W., Friedman A.J., Burrows D.G., Maynes O., Pearce R.W., Wigren C.A., Bogar R.G., Standard
Analytical Procedures of the National Oceanic Atmospheric Administration (NOAA), National Analytical Facility, 19851986:
Extractable Toxic Organic Compounds, NOAA Tech. Memo. NMFS F/NWC-92, 1985, p. 121.
[14]. Ibadov N.A., Huseynov V.I., Suleymanov B.A. (2004) Determination of Polynuclear Aromatic Hydrocarbons By High
Performance Liquid Chromatography, Journal of the Chemical Problems, 2, pp.40-48.
[15]. Reddy Christopher M. And Quinn James G. (1999), Marine Pollution Bulletin, Vol. 38, No. 2, pp. 126-135, 1999.

122

The Impact of Oil Contamination on Soil Ecosystem

[16]. Hawthorne, S. B., Grabanski, C. B., and Miller, D. J., Measured Partitioning Coefficients for Parent and Alkyl Polycyclic
Aromatic Hydrocarbons in 114 Historically Contaminated Sediments: Part I, Koc Values, Environmental Toxicology and
Chemistry, 25, 2006, pp. 2901-2911.
[17]. Thomas J. McDonald, Bo Wang, Susanne J. McDonald and James M. Brooks. Quantitative Determination Of Aromatic
Hydrocarbons Using Selected Ion Monitoring Gas Chromatography/Mass Spectrometry TDI-Brooks International./B&B
Laboratories Inc. College Station, Texas 77845.
[18]. UK- A guide to practices, procedures and methodologies following oil spill contamination incidents (2004).
[19]. Douglas, G. S., McCarthy, K. J., Dahlen, D. T., Seavey, J. A., Steinhauer, W.G., Prince, R. C. and Elmendorf, D. L. (1992).
The use of hydrocarbon analyses for environmental assessment and remediation. Journal of Soil Contamination 1, 197-216.