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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 spi lled 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).

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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 C211 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 (C19) 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 Gulkin.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.

~: :. F ") F A SEA

.•, • ,..a,.._ . ";-..

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 capillary 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 carrier gas (29.4 psi press.)

Results

The standard curve of Nassia cargo 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: hexane mixture used in the second and third surveys. In the first survc) the highest pollution was determined ell 83 station.Kl 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 highest 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 .,.------------

_./ I .../ ______ _j _,/ I 80 -l------------------------

I 60 J

J /

/- · ; __ ---·--·--]' /' 40 ~~-----------

1 20 - _____..- /,r/ ---- ------ __ _J I

/ __ ;-- I ·/ 0 -· -----~--_._...,.-·----r----..,...----- ·----r --,-0 2 4 6 8 1 0 1 2 14 1 6 18 20

L . . . (JJg / ;,.,u I -- ---- -- ------ ------------- -- --- ----- --- - ____ I

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 memory.

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 tanker Nassia.

Table. I UVF results of mussels samples(f..lg/g-dry weight)

~ Nl N12 N3 . v

Kl 15 82 57 81 5 15 I)

82 18 69 250 /JJ 60 1-l4 53

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Fig. 3 The chromatogram of aliphatic fraction of mussel in station Kl (a) and its spectrum. Pr : b I. Ph : b2 .

............ '"""' pr

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-"!-.!2.. _ _ _L_ Ji __ fl _ '!ll 190 -l!L.!lll. -'19 .l"l. .• \20 - 'lil !'0 1!9_ !W .. P>_ l!O_J.O ~-~ •. ---..li'L I.J : :id.<.OttC•ne, 2, o,lO, 14• l e U..at:thyl---- ----------- -. - ----- .w-IH7aa.l12awij KIWJI1A&.DII- ---··--· ---

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72

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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 fh

a

: Pentad~c~n~. 2,o,10,14-t~tr•muthyl-

71

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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.

E I -

I:: ...... ""'"" ....... ""'"" ....... ..... ...... -

nc huiiillb

a

..

__ 1Cl0 __ 10.0D_1_2.C0~4GL_!I_00 __ 11CIO__!O.OO_p_D0__]4~21)XI_21CO_JOQP_J2.00_)40D_li.OD._JBC'Cl~.eG(I)~~-44(1)

J[' l i " _,.,,.....,.,..,,-.,--·---------·--·--·----,

~l.LJ ,__lu.L,. ;_J,~~r _:~_::__:;c . .,..,..;.~·~'"-'"-"-"' "-"'-"''"-"'--_:,.II . .~ __ 40~_!0_!t5_.,_ t5_7tl_75._fl0_ !1! ., ~ _1r'01~1101 ,, 110 1 251~ 1~.]40 ,_ .. , tfO '"'1101fl51}0l7&1801851!101~2t'X11m210:'!!2:!02'1!DJ~7t0245~:""'"<'52"...0i'e"\ i'o--- - : Hexadec.!lne, 7. , 6,10, 14-tetramethyl-

~ ~t11A"15i«'*"*'t~Si.AL01.;------- l

: ~~I ~,J_j_3·~"~~"',r., ·~, ~, ... ·~ _::_ _·r ~,._m_ •• ,.~_,,. _ ~: ~~-~'0_!10 __ to __ 10 __ 81. to_ 100 .• 11t'. _120. IXI _,IC'> , _ IYI leo') _ ti'O __ 11!10 190_ X'O 110 no 110 2ot0 ~ ~ 77t1 1"\0

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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-

.. bl

II 1

.. 1 'l'i' 121

I I 11 105 I ... I 141 '" 18) 183

!,,, .~ 1 tl, .•ZIIr ,,J,JIJ •"''~ ,,1, l,,,l,.,hli li 1;J;I.,,!,.).:;.,~"L'r1,./,,,.',~.···:~ •• ,l~ 80 7'0 ell to . 100 . 110 1M 130 140 150 UIO ITO 180

'""" ., .. ~1, .... ~.:, JO .. ..

ID fleKadecane. 2, 6, 10, 14-tetramethyl-

-- --·- ""'..,;dlii.e5i02iij;ri\: U...S.W..DH-----

0000 ., ... b2

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Fig. 7 The chromatogram of aliphatic fraction of Nassia crude oil (a) and its spectrum.

76

Pr : b I. Ph : b2.

U.l

... I

Pent3.decane, '2, S,lO, t-4·t'!tr3metl':yt -

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b2 I

Fig. 8 The spectrum Pr : bl, Ph: b2 taken from HP memory.

nee

900: 1 &a) I 70'.Xl l 600)]

I

50001

40011 30:Xll

137470 Pe'lla~e:a"l<' . 2.6.10,14·16lr•melhyl-

113

~i...J .!,.I...J. ... l ..1. .. .J. J .. T .T :r. ~~ J . ~·~ .. ~.· .. .. ~.~, : ' -> 20 30 .o10 so SJ 10 eo oo 100 110 120 130 1-40 1so 160 no 180 100 200 210 220 230 2-40 250 260 vo

l 5

8000 I 7000 1

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29

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77

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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 14-238 p.g/g. in Karadag trace and Danube region 350 p.g/g. Kerchensky Strait 16 ~tg/g (Shechekaturina 1991). The 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~malarda degi~ik istasyondan altnan su. sediment ve deniz canltlan iizerinde ytgt~an 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 saptannu~ttr . Bu yaymda midye iizerinde ytgt~an petrol kirliligine ait yalmz alifatik fraksiyon ele altnmt~ttr. Bu analiz pristane/phytane oramna dayandmhm~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~an petrol arttgt miktanntn Karadcnit.de ba~ka

ara~tmctlar taraftndan yaptlan c;:ah ~malarda tespit cdilen miktardan (Tuna haric;:) yiiksck oldugu saptanmt~ttr.

Aknowtcgment

The.: author~ thanks to I'.J.C.Tihbc.:tts (M.Scan Ltd. ) and B. Dicks (blt . Tank.OW11. Pol. Feu. Ltd.) lor kino help.

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References

A11el. M. (1984) Petroleum hydrocarbon pollution in the Rijeka Bay determined by Fluorescence spectroscopy and gas chromatography. VII Journees etud. Pollution. Lucerne. C.I.E.S.M.

Ehrhardt. G.M .. Bums. K.A. (1992). Hydrocarbons and related photo-oxidation products in Saudi Arabian Gulf Coastal Waters and hydrocarbons in underlysing sediments and bioindicator bivalves. /1/or.l'oll. Bull. 27.pp. 187-197.

Goldberg. E.D. (1976) The health of the oceans. The Unesco Press. Paris. p.ll9.

Lee. R.F .. Sauerhcber. R. and Benson. A.A. (1972) ,\'cience 177.pp. 344-346

Mironov. O.G. (1991) Hydrocarbons in the Black Sea ecosystem. ln Proceeding of the Black Sea Symposium (K.C.Gliven. ed.). published by Karadeniz Egitim. Kliltlir ve <;evre Yakf1. Acar Matbaas1 199-t pp. 92-100.

Morris .. R.J.and Gulkin. F. (1975) Hydrocarbon. in the marine environment. In : Environmental Chemistry vol. I. The Chemical Society .. Burlington House. London. pp.81-108

NAS ( 1975) Petroleum in the marine environment. Washington. D.C .. National Academy of Science p. I 07

Ogiizlilgen. S. (1995) The importance of pilotage services in the Turkish Straits for the protection of life property and the environment. In Turkish Straits. New problems. New solutions (1. Soysal. ed.). Isis Ltd. Beylerbeyi. istanbul.

Shechekaturina. T.L.( I991 ). In Polikarpov. G.G .. Zaitsev. Yu.P .. Zats. V.l... and Radchenko. L.A. Pollution of the Black Sea (Levels and Sources) ln : Proceedings of the Black Sea Symposium 16-18 Sept. 1991 Istanbul. (K.C.Giiven. ed.). Karadeniz Egitim. Kiiltiir ve <;evre Koruma Vak.fl. Acar Matbaas• 1994. pp. 15-42

Tibbetts .. P.J.C. and Large R. (1988) Improvements in oil Fingerprinting: GCIHRMS of sulfur heterocycles. Petronnn~vsis 87. 45-57.

Received 24.10 .. 1995

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