Contaminants in oils and fats:

analysis
l i andd regulations
l ti
Florence LACOSTE
l COS
ITERG
f lacoste@iterg com
f.lacoste@iterg.com

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013 1

 ITERG
A synergy of competences services provided

Quality of Safety of
products
d t
Industrial environment
and Safety
• Analytical Controls
• Production, made‐to‐order refining
• Research and Development
Green chemistryy • Consulting, audit
Consulting audit
And process
• Watch and Information
• Professional Training

French CTI (Technical Industrial Center) 

Technology Nutrition, Metabolism
Authorityy : French Ministryy of Industryy
R fi i
Refining and
d Health
H lth
Staff : 80 
2012 Budget : 6M€
ISO 9001 V 2008 & ISO 17025

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 Contents
Heavy metals
Regulation ?
Polycyclic aromatic hydrocarbons
Polycyclic aromatic hydrocarbons
Recognized analytical methods ?
Mineral oil
Risks ?
Phthalates
Regulation Methods Risks

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 Origins of undesirable compounds

Environmental contamination
Environmental contamination Crop protection
i
metals, dioxins & PCBs, PAHs,  pesticide residues, mineral oil
mineral oil

Production process
Transport & storage
& storage PAHs, mineral oil, phthalates, 
PAHs, mineral oil, phthalates,
phthalates, mineral oil  3‐MCPD esters & glycidol esters

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 Heavy metals

Environmental contamination (air, soil) 

Regulation & International Food Standard

 Codex Standard 193‐1995 

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

Regulation Methods Risks

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 Metals : methods & regulations
graphite furnace Analytical methods
Hollow cathode
ISO 8294  Cu, Fe, Ni 
ISO 8294    C F Ni
graphite furnace atomic ISO 15774  Cd
Prism absorption ISO 12193  Pb

inductively coupled plasma  ISO/TS 21033  Cd, Pb, 

optical emission
optical emission , ,
Cu, Fe, Ni

METALS Cd Pb Ni As Cr
LOQ  atomic absorption 
(mg/kg) 0,002 0,010 0,015 0,010 0,005

Regulation limit for oils ‐ 0,10

0 10 02
0,2 0,1
0 1 0,05
0 05
(mg/kg) EU&Codex France Codex France

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 Metals : occurrence in edible
oils & fats
Lead may be present in some cold‐pressed oils
Lead Cadmium

20 %

ITERG  data (1992 ‐1999)

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 Heavy metals transfer from soil
to rapeseed oil
Metals in the seeds are transferred to meals after seed crushing
Cd-rich urban sludge
Culture
Cu-rich urban sludge

0,09 mg/kg Cadmium

oilseeds
o seeds
3 09 mg/kg
3,09 /k CCopper

Drying , Crushing,
Extraction
ADEME Project – 1999/2001

meall crude
d oil
il
PROLEA
CETIOM

0,17 mg/kg Cadmium < 0,002 mg/kg Cadmium

6,13 mg/kg Copper 0,02 mg/kg Copper

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 Polycyclic
y y aromatic
hydrocarbons

Environmental contamination

Production process

Regulation

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

Regulation Methods Risks

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 PAHs origins in vegetable oils
Oilseeds
Extraction Crude oil
+ * Grapeseed oil
HAP * Coconut oil
v * Pomace olive oil
*  Sunflower oil

A ti carbon
Active b
PAHs Deodorisation

Light PAHs

Heavy PAHs

Refined oil
Drying (+  HAP)

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 Structure of PAHs

naphthalene acenaphthene acenaphthylene fluorene phenanthrene anthracene

GT GT

LIGHT PAHs

fluoranthene pyrene benz(a)anthracene chrysene

GT GT GT GT

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

indeno(1,2,3‐cd)pyrene

GT GT
HEAVY PAHs

GT : genotoxic
dibenz(ah)anthracene benzo(ghi)perylene

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 PAHs regulation: (EC) n°1881/2006
contaminants in foodstuffs
Maximum  Maximum  Maximum 
PAHs levels  oils and  levels  coconut  levels cocoa 
fats (µg/kg)
fats (µg/kg) oil (µg/kg)
oil (µg/kg) butter (µg/kg)
butter (µg/kg)

Sum of 4 PAHs: 35,0

‐ benzo[a]pyrene from 1.4.2013
from 1.4.2013
‐ benz[a]anthracene 10,0  20,0 until 31.3.2015
‐ benzo[b]fluoranthene 30,0
‐ chrysene
h from 1.4.2015 

5,0
benzo[a]pyrene 2,0 2,0
from 1.4.2013

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 PAHs: determination methods
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 xxxx (JRC) : 4 PAHs in foodstuffs  (SEC + SPE + GC/MS)

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)

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 PAHs: some difficult matrixes
ISO 15753
ISO 15753 :: sunflower
sunflower oil (1 µg/kg)
oil (1 µg/kg)
: reference solution (5 µg/kg)
(1) NA
(2) AC

11
(3) FL LU

5
200

12
(4) PHE

9
175

8
4
(5) ATR

7
150

14
4
0
10
2
125
(6) FA

13
100

(7) PY 75

15
6
50
(8) BaA
25

(9) CHR 0
0 5 10 15 20 25 30 35 min

(10) BbFA
(11) BkFA LU
: refined pomace olive oil
(12) BaP
: reference solution (5 µg/kg)
1000

(13) DBahA
800

(14) BghiP
600
(15) IP

11

12
400

13
5
3

14
8

10
4

7
6

200

15
1

0
0 5 10 15 20 25 30 35 40 min

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 Edible oils: PAH4 Levels
benzo(a)pyèrene
chrysène
Screening 2012 : 21 vegetable oils benz(a)anthracène
( )
benzo(b)fluoranthène
12
EU limit: 10 µg/kg
10

8
PAH4 en µg/kg
6

ITERG  data (2012)

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 Mineral oil
Environmental contamination (air, soil)
contamination (air soil)
Crop protection
Transport & storage
Transport & storage
Production process
Regulation
R l ti
 (EC) n°1151/2009 ‐ import of sunflower oil from Ukraine
 EFSA Scientific Opinion on Mineral Oil Hydrocarbons in food (2012)
EFSA Scientific Opinion on Mineral Oil Hydrocarbons in food (2012)

Regulation Methods Risks

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 Mineral oil composition

EFSA Scientific Opinion, 2012

Mineral oil: complex mixture of hydrocarbons
 MOSH:
MOSH straight
i h or branched
b h d alkanes
lk & alkylated
lk l d cycloalkanes
l lk
 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)

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 Mineral oil detected in vegetable oils
 2008  contamination of sunflower  Natural
hydrocarbons
oil from Ukraine with a mineral oil
oil from Ukraine with a mineral oil 
from unknown origin

 2009  contamination of walnut oil  Mineral oil :

with a food grade lubricant oil during  unresolved
complex mixture
refining process
refining process

 2010  identification of compounds 
eluted as mineral oil in grapeseed oils
eluted as mineral oil in grapeseed oils

 2011  contamination of milk fat 
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
with a food grade lubricant oil during 
ih f d d l bi il d i
Contaminated Sunflower oil (MOSH analysis)
production

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MOSH determination: ISO/ CD 17880
Fractionation of the sample by liquid chromatography on 
silica gel or silica gel impregnated with AgNO3
Q
Quantification with an internal standard (C18 or C20)
ifi i ih i l d d (C18 C20)
GC/FID analysis on an short apolar column
Vide
Vid
18,5 g  Silicaa gel

1 g  oil

On‐column injection (2 µl)
40°C DB5 HT (15 m‐0,25 mm‐0,1 µm)

Elution with hexane Solvent evaporation GC analysis

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 MOSH determination: critical points
 Cross contamination to be avoided
 Quantification of C10 to C50 without loosing the volatile hydrocarbons
Quantification of C10 to C50 without loosing the volatile hydrocarbons
 Analysis of all types of samples (crude, refined, vegetable & animal  oils  & fats)
 Limit of quantification as low as possible
 Integration of the hump & subtraction of  the “natural hydrocarbons”

It is necessary to integrate twice the chromatogram

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 Mineral oil determination:
ISO/ CD 17880 pre-collaborative trial
43 participants from 12 countries (2012)  dispersed results 
50 mg/kg spiked olive oil
50 mg/kg spiked olive oil 450 mg/kg spiked crude soya oil
450 mg/kg spiked crude soya oil
180 600

Lab mean value (mg/kg)
160
n value (mg//kg)

550
140
120 500
100
450
80
Lab mean

60 400
40
350
20
0 300
Laboratories Laboratories
sample olive oil crude soya oil
mean value (mg/kg)
mean value (mg/kg) 62 447
reproducibility (mg/kg) 49 153
Horrat value 3,3 1,9

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 Phthalates
T
Transport & storage
t& t

Production process
Regulation
 EFSA Scientific Opinion on food additives, flavourings, processing aids and 
p g p g
materials in contact with food (2005)
 (EC) n°10/2011 ‐ plastic materials and articles intended to come into contact 
with food
with food

R l ti
Regulation M th d
Methods Ri k
Risks

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 Chemical structures of phthalates
O Di-methyl PHT DMP R1=R2=methyl

C R1 Di th l PHT
Di-ethyl DEP R1 R2 h l
R1=R2=ethyl
O Di-isobutyl PHT DIBP R1=R2=isobutyl
O Di-butyl PHT DBP R1=R2=butyl
C
Di-hexyl PHT DHexP R1=R2=hexyl
O
R2 Benzyl butyl PHT BBP R1=benzyl R1=butyl
Di n heptyl PHT
Di-n-heptyl DHepP R1 R2 heptyl
R1=R2=heptyl
Di-(2-ethyl hexyl) PHT DEHP R1=R2=ethyl‐2 hexyl
DEHP Di-n-octyl PHT DNOP R1=R2=octyl
Oily visquous liquid Di-isononyl PHT DINP R1=R2= isononyl
MW: 390,6 g/mol Di-isodecyl PHT DIDP R1=R2=isodecyl
BP: 385°C
BP: 385 C
Water solubility: 3 µg/l
High affinity for fat (log Kow : 7,5)

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 Phthalates are everywhere

Toys Shoes‐Boots‐Gloves
Car undercoating
Car undercoating
Child‐care articles Out‐door & rainwear
Dashboard‐Door 
panels‐Safety glass

Cosmetics : Perfume
Flooring‐Roofing‐
Flooring Roofing Wall covering
Wall covering Hairspray‐Deodorant Pharmaceuticals
Ph ti l
Adhesives‐Sealant‐Rubber Skin emollient‐nail polish Medical devices : 
Paints‐Shower curtains  fingernail elongators Catheters‐Blood bag
Wires & cables‐Fresheners
Wires & cables Fresheners

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 (EC) n°10/2011 - Plastic materials &
articles into contact with food
Specific migration 
Specific migration
T b
To be used as
d
limit in food
BBP 30 mg/kg
Plasticizer
Pl ti i in single‐use material containing non‐fatty 
i i l t i l t i i f tt
DINP
∑ = 9 mg/kg foods except infant formulae
DIDP
DEHP 1,5 mg/kg Plasticizer  in repeated use materials containing non‐
DBP 0,3 mg/kg fatty foods

 Material containing these phthalates cannot 
be used for oils & fats 

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 DEHP migration in rapeseed oil
Chromatogram Plots t bi )
(PVC tubing)
MCounts 10-03-2009-05-37-56-CO raff.SMS TIC Filtered
50:249

ITERG data, 2008 Rapeseed oil before migration
75
7.5
SPME/GC/MS
SPME/GC/MS 
5.0

2.5

DEHP (DIOP)
+ 23.939 min

26.078 min
21.028 min
17.176 min
16.239 min
11.608 min

13.274 min

DNOP
DHexP
DIBP

DBP
DMP

DEP
0.0

MCounts 10 03 2009 08 10 47 CO raff

10-03-2009-08-10-47-CO ff + ttuyau.SMS
SMS TIC Filt
Filtered
d
50:249

Rapeseed oil after immersion of PVC tubing

23.942 min
DNHepP
7.5

DEHP
5.0

2.5

26.002 min
+ 17.173 min

26.099 min
21.115 min

21.320 min
15.074 min
11.582 min

13.272 min

DNOP
DHexP

DIDP
BBP
DBP
DMP

DBA
DEP

0.0

10 15 20 25 30
minutes

Oil sample kept in a bottling machine tub 
p p g 3 825 mg/kg of DEHP !
g/ g

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 Phthalates: ITERG’s procedure
m/z 111 (adipates)
GC/MS m/z 149 (phthalates)
5 50:500

DNPP
PDMS
DBP
4 DEHP
DHexP
DMP
3
1 g oil DEP
DNOP
DB5ms   2

(30 m; 0,25 mm; 0,25 µm) 1

HS SPME
HS SPME A l i
Analysis 0
7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0
minutes
kCounts
200

Advantages of the method : 150

100 DINP
No contamination, no solvent
50 DIDP
Rapid analysis (45 min) 0
25 26 27 28 29 minute
30
Sensitive LOQ < 0,1 mg/kg (excepted DINP & DIDP) s

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 DEHP in oils & fats
43
5
mg/kg
4

ITERG data, 2009

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 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
• H
However some contaminants
i can be
b formed
f d during
d i theh production
d i
process of vegetable oils such as esters of 3‐MCPD & esters of
g y
glicydol.

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013

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 A special
p thanks to….

ITERG Analysis Department team

COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013
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