·.~.~.~ ••ZAVOD ZA JAVNO ZDRAVSTVO

~DR. ANDRIJA STAMPARINSTITUTE OF PUBLIC HEALTH

Division of Environmentaland Occupational Health

Department of Environmental Protectionand Health Ecology

Zagreb, August 29th 2014

Division of Environmental and Occupational Health

REPORT

on examination of one crude mineral oil sample and two soil samples after treatment withremediation agent for refinery and chemical waste

Sampling

As arranged on June 10th2014 (with Mr. 1. Rogan) one sample of crude mineral oiland two soil samples have been submitted for analysis July 2nd 2014, after treatment withremediation agent for refinery and chemical waste labeled as BCP35S I BioSurf.

Sample preparation and recording

All samples were extracted with an organic solvent (dichlorometane) and analyzedin the combined system of gas chromatograph - mass spectrometer (GC-MS system) in orderto identify all mineral oil constituents and micro-organic contaminants present in treated soilsamples .

. A sample of crude mineral oil was prepared for GC-MS analysis by direct dissolutionin dichloromethane, while the extracts of treated soil samples were prepared using ultrasonicbath and evaporation system under nitrogen stream.

Analysis was performed by direct injection of sample extracts into gas chromatograph.Standard conditions of temperature programming were: 5 min 40°C/1 O°C per minute/280°C21 min, applied ionization method used electron impact energy of 70 eV. Full mass spectra ofthe separated compounds were recorded in the range from 45 up to 450 Da.

Identification of micro-organic contaminants present was conducted by comparing thefull mass spectra with the mass spectra of known compounds from available databases(similarity index SI ~ 85, Wiley, NIST, PMW databases). The presence of components fromthe group of linear and branched saturated hydrocarbons and the components from the groupof lower substituted benzenes (group of BTEX: benzene, toluene, ethyl benzene and xylenes),C3 and C4 substituents was especially tested.

Zavod za javno zdravstvo "Dr. Andrija Štampar" _ Mirogojska cesta 16 _ HR-10000 ZagrebTel. 01/46 96 242; 46 96 240 _ Fax. 01/46 78 015 _ ekologija@stampar.hr _ www.stampar.hrOIB: 33392005961 (MB: 03270661) _ Poslovni račun: 2340009 - 1100159915, PBZ d.d. Zagreb

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Analysis resuits

All of the recorded characteristic profiles for all samples were reviewed and verifiedand the identifications of the present components were made.In the crude mineral oil sample used for contamination of soil samples, linear and branchedhydrocarbons with CIO to C56 carbon atoms in the molecule dominated, with approximateshare of 93%. Identified substances from the BTEX group (benzene, toluene, ethyl benzeneand xylenes) were determinated with approximate share of 3.5%, further, identified C3-benzene substituents comprised approximately 2.5% and identified contents of the group C4substituents of benzene made the remaining 1% in the total mass of raw mineral oil(Appendix 1).

After the review of the profiles of the recorded soil sample extracts, treated with theremediation agent, none of the extract samples contained ingredients identified like those inthe analyzed solution of crude mineral oil (Appendix 2 and 3). Although, the presence of onlytraces of some crude mineral oil components was indicated. This may be the result of usedsolvent contamination.

Conclusion

Test resuIts indicates a very high efficiency of remediation agent for refinery andchemical waste (labeled BCP35S / BioSurf).

Appendix

Characteristic profiles of TlC - chromatogram with mass fragments for a group ofalkanes and for the groups of lower benzene substituents.

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Appendix 1. Waste Abr.346std1- btex 1uL/10mL = 20 mg = 2ug (= 30min SO°C)

TIC

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0.5

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Appendix 2. Waste - soi! Abr.347 - btex 1uL/10mL = o,5g

40°C 5'/10°C/ 280°C 21'_Pc=1,45 mLHe/min _Ov=1,45 kV BTEX3start=4,6'

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m/z 78 Pk = 54,7

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Appendix 3. Waste - soil Abr.348 - btex 1uL/lOmL = l,Og

40°C 5'/10°C/ 280°C 21'_Pc=1,45 mLHe/min _Dv=1,45 kV BTEX3start=4,6'

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2.

1.

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m/z 78 Pk = 142

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m/z 91 Pk = 94 m/z 120 Pk = ND m/z 134 Pk = 9,3

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