Contaminants in oils and fats: analysis and regulationsaocs.files.cms-plus.com/Meetings/ChinaPresentation/Pa… · · 2014-12-011 AOCS-CCOA Joint Symposium on Functional Lipids

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AOCS-CCOA Joint Symposium on Functional Lipids Shanghai, China on 19-20 November 2014

Florence LACOSTE ITERG (Pessac, France)

[email protected]

Contaminants in oils and fats: analysis and regulations

AOCS-CCOA Joint Symposium on Functional Lipids, Shanghai, Nov 2014

Guillaume

CHANTRE

Technical

Industrial

Center

French

Ministry

of Industry

80

Staff

6 M€

(turnover)

ISO 9001

ISO 17025

Presentation of ITERG

2


Polycyclic aromatic hydrocarbons

Mineral oil

Phthalates

3-MCPD esters & glycidol esters

Contents

3

Origins ?

Regulation ?

Recognized analytical methods ?

Risks ?

Risks Methods Regulation


Polycyclic aromatic

hydrocarbons

4

Environmental contamination

Production process

Regulation

(EC) n°1881/2006 – contaminants in foodstuffs



PAHs origins in vegetable oils

PAHs

Extraction

v

Oilseeds Crude oil

+

HAP

Active carbon

Heavy PAHs

Light PAHs

Deodorisation

Drying

Refined oil

(+ e HAP)

5

* Grapeseed oil * Coconut oil * Pomace olive oil * Sunflower oil

AOCS-CCOA Joint Symposium on Functional Lipids, Shanghai, Nov 2014 6

benzo(b)fluoranthene benzo(k)fluoranthene benzo(a)pyrene indeno(1,2,3-cd)pyrene

dibenz(ah)anthracene benzo(ghi)perylene

HEAVY PAHs

naphthalene

acenaphthene acenaphthylene fluorene phenanthrene anthracene

fluoranthene pyrene benz(a)anthracene chrysene

LIGHT PAHs

Structure of PAHs

GT

GT

GT GT GT

GT : genotoxic

GT GT

GT


PAHs regulation: (EC) n°1881/2006

contaminants in foodstuffs

PAHs Maximum

levels oils and fats (µg/kg)

Maximum levels coconut

oil (µg/kg)

Maximum levels cocoa

butter (µg/kg)

Sum of 4 PAHs: - benzo[a]pyrene - benz[a]anthracene - benzo[b]fluoranthene - chrysene

10,0 20,0

35,0 from 1.4.2013 until 31.3.2015

30,0 from 1.4.2015

benzo[a]pyrene 2,0 2,0 5,0

from 1.4.2013

7


PAHs: determination methods

8

Method ISO 15302 ISO 15753 ISO 22959

BaP LOQ (µg/kg) 0,1 0,2 0,1

Reproducibility (CVR%) 27 % (2,1µg/kg) 41% (3,21µg/kg) 10 % (2,6µg/kg)

Principle

PAH isolation: liquid chromatography (alumina, silica gel,C18-silica gel) or HPLC (donor-acceptor complex chromatography, size-exclusion chromatography)

Analysis: HPLC/fluorescence or GC/MS

ISO 15302 : benzo[a]pyrene in oils (LC + HPLC/FLD)

ISO 15753 : 12 PAHs in oils (2 SPE + HPLC/FLD)

ISO 22959 : 17 PAHs in oils (DACC on-line + HPLC/FLD)

EN 16619 : 4 PAHs in foodstuffs (SEC + SPE + GC/MS)


0

10

20

30

40

50

60

70

80

90

100

<0,5 0,5 - 1 1 - 2 2 - 5 5 - 20 20 - 50 50 - 100 >100

% s

amp

les

Benzo[a]pyrene (µg/kg)

Virgin olive oil

Olive oil

Pomace olive oil

Grapeseed oil

Sunflower oil

EU limit

2 µg/kg

BaP : occurrence in vegetable oils

9

EU Scoop Task (Oct 2004)


Edible oils: PAH4 Levels

Screening 2012 : 21 vegetable oils

PAH4 en µg/kg

benzo(a)pyrene chrysene benz(a)anthracene benzo(b)fluoranthene

ITERG data (2012)

0

2

4

6

8

10

12EU limit PAH4 : 10 µg/kg


Mineral oil

11

Environmental contamination (air, soil)

Crop protection

Transport & storage

Production process

Regulation

(EC) n°1151/2009 - import of sunflower oil from Ukraine

EFSA Scientific Opinion on Mineral Oil Hydrocarbons in food (2012)



EFSA Scientific Opinion, 2012

Mineral oil: complex mixture of hydrocarbons

MOSH: straight or branched alkanes & alkylated cycloalkanes

MOAH: aromatic hydrocarbons including alkyl-substituted

Hydrocarbon compound number in mineral oil > 100 000 for those with less than 20 carbon atoms !

Different products & composition: diesel fuel, white oil, lubricant …

Technical grade mineral oil contain 15-35 % MOAH, which is minimised in food grade MOSH (white oils)

Mineral oil composition

12


Minutes

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30

Millivo

lts

50

100

150

200

250

300

350

400

TRACE GC-Channel 1

sample 7

Name

13

Minutes

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30

Millivo

lts

50

100

150

200

250

300

350

400

TRACE GC-Channel 1

sample 7

Name

Natural hydrocarbons

Mineral oil : unresolved complex mixture

EI

Mineral oil detected in vegetable oils

2008 contamination of sunflower oil from Ukraine with a mineral oil from unknown origin

2009 contamination of walnut oil with a food grade lubricant oil during refining process

2010 identification of compounds eluted as mineral oil in grapeseed oils

2011 contamination of milk fat with a food grade lubricant oil during production


Fractionation of the sample by liquid chromatography on

silica gel impregnated with AgNO3

Quantification with an internal standard (C18 or C20)

GC/FID analysis on an short apolar column

MOSH determination: pr ISO17780

14

18

,5 g

Sili

ca g

el

Elution with hexane Solvent evaporation

Vide

40°C

GC analysis

On-column injection (2 µl) DB5 HT (15 m-0,25 mm-0,1 µm)

1 g oil


pr ISO 17780 validation

15

Mineral oil – 50 mg/kg spiked olive oil sample 2-2013 sample 4-2014

Spiking level (mg/kg) 50 50

Mean (m) 52 52

Repeatability limit (r) 19,3 15,3

Reproducibility limit (R) 51,7 28,1

Horrat value 4,1 2,2

0

20

40

60

80

100

120

140

160

26 3 8 23 21 34 15 31 1 19 4 33 36 9 28 17 30 27 29 24 11 10 35 5 32 12 6 22 2 18 20 14

Lab

me

an v

alu

e (

mg/

kg)

Laboratories

Sample 2: 50 mg/kg spiked olive oil

0

20

40

60

80

100

120

140

160

20 12 14 10 7 9 13 16 15 1 11 19 4 8 5 2 6 17

Lab

me

an v

alu

e (

mg/

kg)

Laboratories

Sample 4: 50 mg/kg spiked virgin olive oil

Method applicable from 50 to 1000 mg/kg


Phthalates

16

Transport & storage

Production process

Regulation

EFSA Scientific Opinion on food additives, flavourings, processing aids and materials in contact with food (2005)

(EC) n°10/2011 - plastic materials and articles intended to come into contact with food


http://images.google.fr/imgres?imgurl=http://www.prodi-services.com/famille/pis.plb.pvc.tub.jpg&imgrefurl=http://www.prodi-services.com/pis/plb/pvc/index.php&h=130&w=130&sz=8&hl=fr&start=57&um=1&tbnid=2JDRr7keTGj9SM:&tbnh=91&tbnw=91&prev=/images?q=tuyau+pvc&start=40&ndsp=20&um=1&hl=fr&sa=N


Chemical structures of phthalates

Di-methyl PHT DMP R1=R2=methyl

Di-ethyl PHT DEP R1=R2=ethyl

Di-isobutyl PHT DIBP R1=R2=isobutyl

Di-butyl PHT DBP R1=R2=butyl

Di-hexyl PHT DHexP R1=R2=hexyl

Benzyl butyl PHT BBP R1=benzyl R1=butyl

Di-n-heptyl PHT DHepP R1=R2=heptyl

Di-(2-ethyl hexyl) PHT DEHP R1=R2=ethyl-2 hexyl

Di-n-octyl PHT DNOP R1=R2=octyl

Di-isononyl PHT DINP R1=R2= isononyl

Di-isodecyl PHT DIDP R1=R2=isodecyl

17

C

C

O

O

O

OR1

R2

Oily visquous liquid

MW: 390,6 g/mol

BP: 385°C

Water solubility: 3 µg/l

High affinity for fat (log Kow : 7,5)

DEHP


Flooring-Roofing- Wall covering Adhesives-Sealant-Rubber

Paints-Shower curtains Wires & cables-Fresheners

Cosmetics : Perfume Hairspray-Deodorant

Skin emollient-nail polish fingernail elongators

Car undercoating Dashboard-Door

panels-Safety glass

Shoes-Boots-Gloves Out-door & rainwear

Toys Child-care articles

Pharmaceuticals Medical devices :

Catheters-Blood bag

Phthalates are everywhere

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http://images.google.fr/imgres?imgurl=http://www.levillagedenoel.com/cadeau-nounours-noel-cadeau_gif.gif&imgrefurl=http://www.levillagedenoel.com/coloriagescadeauxdenoel.htm&usg=__R2gXzpGJf1yt1ZyclMSoIS3_TKU=&h=400&w=463&sz=7&hl=fr&start=28&tbnid=RWXfJxnhdYT3JM:&tbnh=111&tbnw=128&prev=/images?q=cadeau+de+noel&gbv=2&ndsp=20&hl=fr&sa=N&start=20


Specific migration limit in food

To be used as

BBP 30 mg/kg Plasticizer in single-use material containing non-fatty foods except infant formulae

DINP ∑ = 9 mg/kg

DIDP

DEHP 1,5 mg/kg Plasticizer in repeated use materials containing non-fatty foods DBP 0,3 mg/kg

(EC) n°10/2011 - Plastic materials &

articles into contact with food

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Material containing these phthalates cannot be used for oils & fats


Phthalates: ITERG’s procedure

Advantages of the method :

No contamination, no solvent

Rapid analysis (45 min)

Sensitive LOQ < 0,1 mg/kg (excepted DINP & DIDP)

HS SPME

1 g oil

Analysis

DB5ms (30 m; 0,25 mm; 0,25 µm)

PDMS

m/z 111 (adipates)

m/z 149 (phthalates) GC/MS

DEHP DHexP

DNPP DBP

DEP

DMP

7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 minutes

0

1

2

3

4

5 50:500

DNOP

25 26 27 28 29 30 minute

s

0

50

100

150

200 kCounts

DINP

DIDP


0

1

2

3

4

543

DEHP in oils & fats

ITERG data, 2009

mg/kg

21


3-MCPD esters & glycidol esters

22

Production process (deodorisation)

Regulation for free 3-MCPD

(EC) n°1881/2006 – contaminants in foodstuffs (20 µg/kg in soya sauce)

Regulation for 3-MCPD esters

EFSA statement – related to 3-MCPD esters (2008)

EFSA Scientific Report on 3MCPD occurrence in food in Europe (2013)



H2C

H2C

HC

O

AG

H2C

H2C

HC

O

OH

H2C

H2C

HC

OH

OH

Cl

H2C

H2C

CH

AG

AG

Cl

H2C

H2C

CH HO

Cl

AG

« free » 3-MCPD

Glycidol

3-MCPD monoesters « bound »

Glycidol esters (GEs)

3-MCPD diesters « bound »

49 7 14

3-MCPD esters & glycidol esters Structure

23

Hrncirik, OVID meeting, 2011

7 fatty acids

C12 - C14 C16 - C18

C18:1 - C18:2 C18:3


3-MCPD esters & glycidol esters Analysis strategy

24

Direct methods Indirect methods

Pros No chemical reaction Individual quantification of species

Standards available Good sensitivity

Cons Standards not available Expensive tools: HPLC/MS/MS Purification step needed

Reactivity of the species Global quantification of species

Standards AOCS-JOCS Cd 28-10 for GEs From Masukawa et al., JAOCS, 2010

AOCS Cd 29a-13 for 2- and 3-MCPD & GEs From Ermacora & Hrnirick, JAOCS, 2012

AOCS Cd 29b-13 for 2- and 3-MCPD & GEs From Kuhlmann, EJLST, 2011

AOCS Cd 29c-13 for 3-MCPD & GEs From DGF C-VI 18, 2010 & Weisshar, 2008


Esters de 3-MCPD & glycidol

• Comparable results for the 3 indirect methods • In general, direct methods agreed with indirect methods • Methods did not give reliable results for concentrations < 1 mg/kg

2013 AOCS Collaborative Study

• Palm oil sample, not spiked. • All indirect methods were acceptable

• RBD Canola sample, not spiked. • Variable results between methods


Occurrence of 3-MCPD & glycidol esters in

vegetable oils

26

Screening ITERG 2013

0.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

9.0

10.0

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol b

rute

H. t

ou

rnes

ol r

affi

née

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

3MCPD (mg/kg)

sunflower

grapeseed

0.0

0.2

0.4

0.6

0.8

1.0

1.2

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. c

olz

a

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol

H. t

ou

rnes

ol b

rute

H. t

ou

rnes

ol r

affi

née

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

H. p

épin

s d

e ra

isin

GLYCIDOL (mg/kg)

sunflower

rapeseed

grapeseed

rapeseed

Industrial specidifcation 3MCPD = 2 mg/kg Industrial specification glycidol = as low as possible


Regulation, Methods, Risks

27

PAHs

Mineral oil

Phthalates

3-MCPD esters & glycidyl esters Risks Methods Regulation





Conclusions

• Research of contaminants is part of multiple controls conducted by fat and oil industry to fulfill the EC regulation n°1881/2006.

• In the absence of regulation, the detection of contaminants must be addressed in partnership with authorities according to the toxicity of molecules.

• Risks are rather limited due to the efficient elimination during oil-refining steps

• However some contaminants can be formed during the production process of vegetable oils such as 3-MCPD & glycidyl esters.

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A special thanks to….

ITERG Analysis Department team

Documents

Contaminants in oils and fats: analysis and regulationsaocs.files.cms-plus.com/Meetings/ChinaPresentation/Pa… · · 2014-12-011 AOCS-CCOA Joint Symposium on Functional Lipids