KASIM C.GOVEN, SELMA ONLO, ERDOGAN OKU$ and

Turkish J.Afnr.S'ci. 1.(113):67-79 (1995)
'
OIL CONTAAJINATJON OFM>TILUS GALLOPROVJNC'JALJS
AFrER THE NASSJA ACCIDENT
NA.\:\1A TANKER KAZA.S1NDAN SONRA AfYTILUS
GALLOPROVJNCJALJS''iN PETROL iLE KONTAMjNASYONU
KASIM C.GOVEN, SELMA ONLO, ERDOGAN OKU$ and ERTUGRUL DOGAN
Institute of Marine Sciences and Managemept.University of Istanbul. 3-U70 Vefa.
Istanbul, Turkey
Key words : Oil pollution. Nassia accident. mussel, UVF, GC/MS analysis
Abstract
Oil accumulation in mussels was investigated after the Nassia oil tanker accident on the
Istanbul Strait by applying UVF and GC/MS analysis methods. Pristane and phytane
ratio was used to distinguish the sources of pollution. Amounts of oil varied from 5 to 250
J.lg/g in three surveys. The analysis results revealed that contamination originated from the
oil spills from the tanker involved in the accident
Introduction
Istanbul Strait has a transit traffic of 35.000 ships per annum on the average. The
accidents occured 30 times in 1990. 23 in 1991. 17 in 1992. 16 in 1993 and 9 in 199~
(Ogiiziilgen 1995).The tanker accidents amounted to 28 out of 205 ship accidents in total
during the period of 1982-1994. The major disaster was on 13 th March 1994 when MIT
Nassia carrying Russian crude oil crashed with MN Shipbroker near the coast of Rumeli
feneri 9akan on the istanbul Strait (Fig. I). 2000 tons of crude oil was spilled causing
severe pollution of the sea and also the ensuing fire contaminated the atmosphere with
fumes more than 4 days.
Numerous reports appear on the eiTect of oil pollution on marine organisms. Mussel.
which is the subject matter in this work, is a filter feeding organism. It accumulates sea
pollution matter. The uptake and discharge of oil by mussel was investigated by Lee eta/.
(1972). Ahcl (1984). Mironov (1991). Ehrhardt and Burns(l992).
67
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Various methods were used for the quantitative and qualitative analysis of oil
contamination. The commonly used method for the determination of oil is UV
nuorospectromctry (UVF). This is based on the determination of PAH of oil in sea water,
sediment or marine organisms and the extinction/emission (ex/em) of extracts were
measured at 310/360 nm.
Aliphatic and aromatic fractions of oil were separated by using colwnn chromatography
(Tibbetts and Large 1988) and analysed by GC or GC/MS spectrophotometry. The
problem was the differentiation of ambiguous petroleum hydrocarbon sources. while some
h)drocarbons could be synthetiLed by marine organisms. Normal alkanes primarily with
odd-numbered carbon. alkanes with 15.17,19 and 21 C atom are synthetized by marine
phytoplankton and C21 to C 211 n- alkanes by Sargassum in the highest
conccntration.Branchcd <llkanes. pristane arc also found in marine
organisms.
Cycloalkancs and cycloalkcnc. carotene and aromatic hydrocarbons arc synthctised by
marine organisms (Goldberg 1976). NAS (1975) gave some guidelines to the differentiation
of petroleum and biogenous hydrocarbons as:
- Petroleum often contains homologous series of hydrocarbdns with even and odd number
of carb011 atoms arc present in nearly the same concentration.
- Petroleum contains more kinds of cycloalkane and aromatic hydrocarbons with alkyl
(mono-tetra· mel hyl) substituted.
- Petroleum contams naphtcno aromatic hydrocarbons and also hctero compounds not yet
reported in marine organisms.
Of the isoprenoid hydrocarbons. pristanc (C 19) 2.6.10.14 tctramethyl pcntadecane(Pr).
occurs in phyto-and L.ooplankton.whilc ph)1ane (CXJ) 2.6JO,l~ tetramcthyl hcxadecanc
(Ph) exists in much lesser amount in marine organisms (Morris and Gu lkin.l975). Their
ratio can distinguish the source, whether natural or due to oil pollution (Blumer 1970).
The ratio can be determined by GC or GCIMS analysis.
The pollution of the sea. sediments and marine organisms of Istru1bul Strait caused by
Nassia tanker accident was investigated by our laboratory (Unpublished data).
This paper reports the part of the pollution studies on the aliphatic fraction of mussels.
Material
Mussel (Aivttlus gnlloprovmcmlis) was taken from stations in Istanbul Strait by diving
(Fig. I). Th~ research was carried out from RN Arar on the dates:
Nr 1: 21-27 Aprill99~.
Nr 2: 5-11 July 199~.
Nr 3: 23-28 Sept. 1994.
UV Fluorospectrophotometcr (Hitachi 650-IOS).
GC/MS (Hewlett Packard. HP 6890 GC System/ Mass Selective Detector).
All chemicals and solvents were HPLC grade (J.Baker).
68
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Methods
1- Extraction and saponification techniques
M-Scan standard procedure was used. lOg samples were extracted with dkhloromethane
(DCM) in L11e first survey and an isopropanol:hexane mixture (80:20) in the second and
l11ird surveys.
Fig. I The sampling stations in the Istanbul Strait. K I : Karabumn. B I : Poyraz,
82 : Altmkum. 83 : 8cykoL. x : Rumcli fencri 9akan .
Kl: To where Nassia tanker has been pulled after the accident.
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The extract was evaporated to a small volume on a rotary evaporator, transferred to a
erlanmayer and saponified with potassium hydroxide solution , then extracted with
hexane. Afler drying with Na2S04, it was filtered and concentrated by rotary evaporation
then the volume was adjusted to 10 ml with hexane .5 ml of the resulting total extract was
analysed by UVF and the other part was applied to silicagel column (8 g silicagcl ,2x25cm
column). The elution was made with pentane and DCM successively.Thus aliphatic and
aromatic fractions were separated and analyzed by GC/MS spectrophotometry.
2- Analysis
2. 1. UVF
A standard calibration curve was plotted by using Nassia cargo sample in concentrations
of 2,5. 10 and 15 Jlg/ml in hexane. The ex/em was measured at 280/330 nm for distilled
fuel oil. 310/360 nm for crude oil and 380/430 nm for some oxidation products of PAH,
(Ehrhardt and Bums 1992). Only 310/360 nm (ex/em) results are given in this paper. Oil
concentration in mussel extract was read from the calibration curve.
2.2. Ge/MS
Ge/MS analyses were run on an HP 6890 capillary gas chromatograph connected to a
Hewlett Packard Mass Selective Dctector(MSD) controlled by an HP ehcmstation.Ge/MS
system equipped with 50mx200J..1m i.d. fused HP PONA (Methyl siloxane) coated gl<lss
ca pillary coltunn.Film thickness is 0.50J..lm.eolumn temperature program is as follows:
.woe hold time 6 min, Ramp l is 10°e I min rate to 280°C hold time 10 min Ramp 2 is
w oe /min rate to 290°e hold time 5 min. Sa mples in n-hexane solution were injected in
splitless mode at 300°C injcktor block temperature. Helium was used for ca rrier gas (29.4
psi press.)
Results
The standard curve of Nassia ca rgo sample is shown in Fig .2.
The UVF analysis results are shown in Table I. The amounts of oil in mussels were low in
the first survey at the stations KI. 81 and 82 whereas in the second survey the levels were
the highest. ln the third survey oil amounts were decreased.The low amounts of oil in the
first survey can be attributed to the lower extraction efficiency of hexane. than those of
isopropm10l: hexa ne mi xt ure used in the second and third surveys. In the first survc) the
hig hest pollution was determined ell 83 station.K l station was to the west of the Black Sea
entrance to the istanbul Strait to where the tanker was pulled after the crash.Thc amount
of pollution on the mussels taken here was as expected. When the results of B 1.82 and 83
situated in istanbul Strait arc compared the hig hest values for oil were found at 83 in the
second survey and at 82 in the third survey.
70
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Fig.2. The UVF standard calibration curve ofNassia crude oil.
Flourescence
100 . , . - - - - - - - - - - - -
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A typical chromCJtogram and spectrum of aliphatic fraction of mussel taken from K I. B I.
82 and BJ stations arc shown in Fig. J-6 abc respectively. Fig. 7 shows the GC/MS
chromotogram and spcctnun of aliphatic fraction obtained from Nassia cmdc oil. Fig. 8
shows the mass spectra of pristane (Pr) and phytane (Ph) taken from HP me mory.
As can be seen in these chromatograms pristanc and phytanc peaks arc clearly visible.
Phytanc peaks are identical with aliphatic fraction of Nassia cmde oil. The spectra of Pr
and Ph obtained from mussels extracts were identical with those taken from MS memory.
This can be taken as a good evidence of oil pollution. originating from the tanke r Nassia.
Table. I UVF results of mussels samples(f..lg/g-dry weight)
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Nl
N12
N3
v
Kl
81
15
82
5
15
I)
82
/JJ
18
69
250
60
1-l4
57
53
71
Fig. 3 The chromatogram of aliphatic fraction of mussel in station Kl (a) and its
spectrum. Pr : b I. Ph : b2 .
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Fig. 4 The chromatogram of aliphatic fraction of mussel in station B l (a) and its spectrum.
Pr: bl. Ph: b2.
- - - - nt IU.tJIA,LQ _ _ _ _
pr
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a
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Fig. 5 The chromatogram of aliphatic fraction of mussel in station B2 (a) and its spectrum.
Pr : b L Ph : b2.
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: Hexadec.!lne, 7. , 6,10, 14-tetramethyl-
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~t11A"15i«'*"*'t~Si.AL01.;-------
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Fig. 6 The chromatogram of aliphatic fraction of mussel in station B3 (a) and its spectrum.
Pr : b I, Ph : b2.
ro
F'entadec!lne, 2, 6. tO. 14-tetntmethyl-
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bl
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Fig. 7 The chromatogram of aliphatic fraction of Nassia crude oil (a) and its spectrum.
Pr : b I. Ph : b2.
Pent3.decane, '2, S,lO, t-4·t'!tr3metl':yt -
U.l
...
I
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76
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Fig. 8 The spectrum Pr : bl, Ph: b2 taken from HP memory.
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lliscussion
Mussels are used in this work to measure the poiJution effect of Nassia Tanker
accident.The highest pollution was determined at station BJ followed by B2. whereas Bl
showed the least polluted. In the first survey the oil amounts in the mussels were found
low in all the tests because hexane was found insufficient as the extraction solvent,whereas
oil content in sea water in the first survey was higher than in the second and third surveys
at the same stations (Unpublished data).
Ahcl (1984) reported that the oil concentration of Mytilus f!.a /loprovincia/is in Rijeka Bay
was around 1.5-19 p.g/g (fresh weight). Aromatic hydrocarbons in mussels from different
regions of the Black Sea were in western Crimea 14-23 8 p.g/g. in Karadag trace and
Danube region 350 p.g/g. Kerchensky Strait 16 ~tg/g (Shechekaturina 1991). T he oil amount
found in mussels at Istanbul Strait after Nassia tanker accident was higher than that
reported at Rijeka Bay. Our results were also higher than those reported for the mussels
of the Black Sea (except Danube).
GC/MS analysis on Pr/Ph ratio proved that oil pollution increased in the mussels ncar the
location of the accident.
Ozct
istanbul Bogazmda mcydana gelen Nassia tanker kazast sonrast yaptlan c;:a lt~mala rda
istasyondan altnan su. sediment ve deniz canltlan iizerinde y tgt~a n petrol kirliligi
tayini enstitiimi.izde ara ~tmlmt ~ttr. Petrol kirliligi miktan midyenin total ekstrakttnda
PAH. UVF analizi ile tayin edilmi~tir. Bu petrol kirliliginin Nassia tanker ka;r.asmdan
oht~tugu GC/MS analizi ile sapta nnu ~ttr. Bu yaymda midye iizerinde ytgt ~a n petrol
kirliligine ait yalmz alifatik fraksiyon ele a ltnmt ~ttr. Bu analiz pristane/phytane oramna
daya ndmhm~ttr. Bu c;:a h ~mada aynca ekstraksiyonda kullamlan solvantn ekstraksiyondaki
rolli de belirtilmi~ tir. Kaza sonrast midyelerde ytgt~<lll petrollin miktan degi~ ik tarihlerde
yaptlan incelemeler sonunda bulunan bulgulara gore belirtilmi~tir. Nassia tanker
ka;r.asmdan sonra midyelcrde ytgt~a n petrol arttgt miktanntn Karadcnit.de ba~ka
a ra~tmctlar taraftndan yaptlan c;:ah ~malarda tespit cdilen miktardan (Tuna haric;:) yiiksck
oldugu saptanmt ~ttr.
degi~ik
Aknowtcgment
The.: a utho r~ thanks to I'.J.C.Tihbc.:tts (M.Scan Ltd. ) and B. Dicks (blt . Tank.OW11. Pol. Feu. Ltd .) lor
kino help.
78
. '·
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79
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