The Uniud Project on mineral oils in olive oils

Prof. Sabrina Moret [email protected]

Department of Agri-Food, Environmental & Animal Sciences University of Udine, Via Sondrio, 2A 33100 - Udine, Italy Multi-company financed PhD

Companies support scientific Advantages for the companies research in the framework of a Ø The Company is updated at least every research PhD in order to deepen and six months on the research conducted in update on a topic of common the area of common interest. interest, each contributing with a Ø The company enters a network that also liberal grant of euros (i.e. 2.500 involves other companies interested in the Euro) deductible annually for three same topic. years. Ø Contact with the academic world can bring Upon reaching a sufficient number of further future opportunities to the company subscriptions, the University (such as participation in joint projects on activates a PhD scholarship on the European calls). topic, also organizing at least half- Ø The Company benefits from tax breaks for yearly dissemination meetings to liberal donations in favor of university stimulate a constant dialectic with the research. business world. Characteristics of the sampling sites

Sample Municipality Distance from road/ Use of Site location Type of site code inhabitants* vehicular traffic pesticide

OF1 -Monastir 37485 Semi-rural 150 m/ medium no OF2 (centre) 221530 Urban 1-5 m/ high no MOSH concentrations (mg/kg oil) in extra virgin olive oils from the market OF3A Monastir (centre) 93306 Urban 1-5 m/ high no EVOO1 EVOO2 EVOO3 EVOO4 EVOO5 OF3B Monastir (centre) 93306 Urban 100 m/ high no MOSH C10-16 1.6 2.4 2.8 3.9 4.3 OF4 42937 Semi-rural 200 m/ medium yes MOSH C16-35 8.7 12.3 6.6 10.4 27.0 OF5A -Monastir 17850 Rural 1-5 m / low yes MOSH >C35 1.2 3.5 0.8 3.3 6.7 OF5B Bekalta-Monastir 17850 Rural 200 m/ low yes MOSH TOT 11.5 18.2 10.3 17.6 38.0 OF5C Bekalta-Monastir 17850 Rural 400 m/ low yes OF6 -Monastir 57111 Semi-rural 300 m / medium no OF7 Menzel-Kamel-Monastir 8432 Rural 300 m / low no OF8 83534 Semi-rural >300 m / low yes * 2014 Tunisian census data MOSH content (mg/kg) of the oil oil extracted from olives both by physical mean (Abencor and with solvent)

Oil extraction OF1 OF2 OF3A OF3B OF5A OF5B OF5C OF6 OF7 Abencor 1.4 2.4 3.0 1.7 1.8 0.0 4.2 3.6 1.6 Solvent (MAE) 3.9 10.5 16.6 5.4 7.8 2.3 17.0 12.0 3.5 Ratio 2.9 4.4 5.5 3.2 4.3 4.0 3.4 2.2

Results of a collaboration research between UniUD and Blauel & Co.

Monitoring environmental impact

Six areas differently exposed to environmental contamination in Greece

B Polluted Uncontaminated D areas areas A E F

C Satellite view of the six area where the olive samples were collected

In each area 2 olive trees were chosen where samples of passive paper filters, soils and olives were collected Environmental contamination: olives

For each area two olive trees were identified to collect samples of olives in 2 periods

The olives were washed with water and after a LLE with Constant and little hexane, the superficial contamination was determined contamination (on average by injecting the concentrated hexane extract into the around 2 mg/kg of oil) was LC-GC apparatus found in olives Washed olives were ground and extracted in a microwave independently on the extractor with hexane ethanol (1:1), the extracted fat was collection site diluted and injected to determine total contamination Superficial contamination was about 1-2% referred to whole olive contamination Environmental contamination: soil

mg/kg dry soil

n -alkanes total soil isoalkanes n -alkanes ratio n -alkanes/isoalkanes carbon range hydrocarbons

A 0,8 4,3 21-45 5,5 5,8 supposed B 1,9 6,4 23-36 3,3 9,0 "clean" areas C 1,0 7,2 21-37 6,9 9,7 D 3,7 6,1 21-37 1,6 11,0 supposed E 1,1 8,2 21-47 7,4 9,8 "dirty" areas F 1,1 1,6 23-35 1,5 3,4 Tests with passive traps Passive traps made of paper filters (without oil and with 30 mg oil) exposed in highly trafficked road to determine the air contamination

NO OIL OIL 30 mg 6 cm 6 cm

2 cm 2 cm

7 cm 7 cm

Area 4 cm oil deposition 4 cm

Area for Areas to tear out for Areas to tear out for analysis blank analysis MOSH MOAH oil no oil blank Passive traps

LC-GC traces of oil extracted from cardboard filters placed for 3 weeks in 6 different sites Monitoring on EVOO CR5 EP1 Oil from different sites in Greece: CR1 ü Crete (CR)

ME2 ü Messinia (ME) EP2 ü Argolide (ANC) ü Epidauro (EP)

LU8 MOSH contamination focused on specific LU4 ranges of carbon chain length Lubricants LU3 LU1 LU2 LU5 LU6 LU7 CR5 EP1 CR1 Pesticides

PE2 ME2 EP2 PE1 C20 LU8

LU4

LU3 LU1 LU2 LU5 LU6 LU7

PE2

PE1 Contamination during harvesting

MOSHC20 in EVOO from different areas in Greece 40,0

35,0

30,0

25,0

kg g/20,0 m

15,0

10,0

5,0

0,0

MOSHC20 Sample CR1 Contamination during transport in jute bags

In the jute bag In the oil (considering 100% transfer) Sample CR9MOSH 3.3 mg/kg 322 mg/kg 10 mg/kg

MOAH

76 mg/kg 2 mg/kg Sample CR1 Sample CR4 MOAH Squalene

Sample CR6 Sample CR 5 Six cultivar from one Italian grove collected at the same period far from sources of environmental contamination

3,5 Distribution of different fractions of MOSH in six cultivar of olives 3,0 0,6 0,5 2,5 0,5 0,8 2,0

0,5 1,5 0,2 2,1 1,8 2,0 MOSH (mg/kg oil)

1,0 1,7 1,2 1,4 0,5

0,4 0,3 0,3 0,0 0,1 0,1 DON CARLO ARBEQUINA GHIACCIOLA ORFANA LECCINO NOSTRANA

n-C10-16 n-C16-20 n-C20-35 >n-35 Three variety were followed during ripening

Evolution of the total fraction of MOSH during ripening

4,0 considering 3 cultivar of olive

3,5 3,4

3,0 3,0

2,5

2,0 1,8 ARBEQUINA 1,7 1,9 DON CARLO 1,5 1,5 LECCINO total MOSH (mg/kg) 1,5 1,3 1,0

0,5

0,0 1414 -ottOct 21-ott 28-23 ott 30 NovOct4-nov 11-nov 18-nov 23 Nov time ripening Time ripening No evident trend was observed during ripening

More data are needed!