Contaminants in oils and fats: l i d l ti
l COS
analysis and regulations
Florence LACOSTEITERG
f lacoste@iterg [email protected]
1COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013
ITERGA synergy of competences services provided
Quality of Safety of d tproducts Industrial environment
and Safety• Analytical Controls
Green chemistry
• Production, made‐to‐order refining
• Research and Development
• Consulting audityAnd process
Consulting, audit
• Watch and Information
• Professional Training
Nutrition, Metabolismd H lth
TechnologyR fi i
French CTI (Technical Industrial Center) Authority : French Ministry of Industryand HealthRefining y y yStaff : 80 2012 Budget : 6M€ISO 9001 V 2008 & ISO 17025
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20132
Contents
Heavy metals
Polycyclic aromatic hydrocarbonsRegulation ?
Polycyclic aromatic hydrocarbons
Mineral oilRecognized analytical methods ?
Risks ?
Phthalates
Risks ?
RisksMethodsRegulation
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20133
Origins of undesirable compounds
Environmental contamination iEnvironmental contaminationmetals, dioxins & PCBs, PAHs,
mineral oil
Crop protectionpesticide residues, mineral oil
Production processPAHs, mineral oil, phthalates,Transport & storage PAHs, mineral oil, phthalates, 3‐MCPD esters & glycidol esters
Transport & storagephthalates, mineral oil
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20134
Heavy metalsHeavy metals
Environmental contamination (air, soil)
Regulation & International Food Standard
Codex Standard 193‐1995
(EC) n°1881/2006 – contaminants in foodstuffs (EC) n 1881/2006 contaminants in foodstuffs
RisksMethodsRegulation
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20135
Metals : methods & regulations
ISO 8294 C F Ni
Analytical methodsHollow cathode
graphite furnace
ISO 8294 Cu, Fe, Ni ISO 15774 CdISO 12193 Pb
graphite furnace atomicabsorptionPrism
inductively coupled plasma optical emission
ISO/TS 21033 Cd, Pb, Cu, Fe, Ni
CrAsNiPbCdMETALS
optical emission , ,
0 050 10 20 10
0,0050,0100,0150,0100,002LOQ atomic absorption
(mg/kg)
0,05France
0,1Codex
0,2France
0,10EU&Codex
‐Regulation limit for oils(mg/kg)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20136
Metals : occurrence in edible oils & fatsoils & fats
Lead may be present in some cold‐pressed oils
Lead Cadmium
20 %
ITERG data (1992 ‐1999)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20137
Heavy metals transfer from soil to rapeseed oil to rapeseed oil
Metals in the seeds are transferred to meals after seed crushingCd-rich urban sludgeCu-rich urban sludge
0,09 mg/kg Cadmium3 09 /k C
Culture
oilseeds3,09 mg/kg Copper
Drying , Crushing, Extraction
o seeds
d ill
ADEME Project – 1999/2001
crude oilmealP R O L E AC E T I O M
0,02 mg/kg Copper < 0,002 mg/kg Cadmium
6,13 mg/kg Copper0,17 mg/kg Cadmium
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20138
Polycyclic aromaticy yhydrocarbons
Environmental contamination
Production process
Regulation
(EC) n°1881/2006 – contaminants in foodstuffs
RisksMethodsRegulation
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 20139
PAHs origins in vegetable oilsOilseeds
ExtractionOilseeds
Crude oil+
HAP* Grapeseed oil* Coconut oil
v HAP
A ti b
* Pomace olive oil* Sunflower oil
PAHsActive carbon
Light PAHs
Deodorisation
Heavy PAHs
Light PAHs
Heavy PAHs
Refined oil
Drying
Refined oil
(+ HAP)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201310
Structure of PAHs
naphthalene acenaphthene acenaphthylene fluorene phenanthrene anthracene
LIGHT PAHs
GTGT
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
GT
benzo(b)fluoranthene benzo(k)fluoranthene benzo(a)pyrene indeno(1,2,3‐cd)pyrene
HEAVY PAHsGTGT
dibenz(ah)anthracene benzo(ghi)perylene
GT : genotoxic
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201311
dibenz(ah)anthracene benzo(ghi)perylene
PAHs regulation: (EC) n°1881/2006 contaminants in foodstuffs
PAHsMaximum
levels oils and fats (µg/kg)
Maximum levels coconut oil (µg/kg)
Maximum levels cocoa butter (µg/kg)fats (µg/kg) oil (µg/kg) butter (µg/kg)
Sum of 4 PAHs:benzo[a]pyrene
35,0from 1.4.2013‐ benzo[a]pyrene
‐ benz[a]anthracene‐ benzo[b]fluorantheneh
10,0 20,0from 1.4.2013until 31.3.2015
30,0‐ chrysene from 1.4.2015
benzo[a]pyrene 2,0 2,0 5,0benzo[a]pyrene 2,0 2,0from 1.4.2013
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201312
PAHs: determination methodsPrinciple PAH isolation: liquid chromatography (alumina, silica gel,C18‐silica gel) orHPLC (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 22959BaP LOQ (µg/kg) 0,1 0,2 0,1
Reproducibility (CVR%) 27 % (2,1µg/kg) 41% (3,21µg/kg) 10 % (2,6µg/kg)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201313
PAHs: some difficult matrixes: sunflower oil (1 µg/kg)ISO 15753 : sunflower oil (1 µg/kg): reference solution (5 µg/kg)
(1) NA(2) AC
ISO 15753
LU
150
175
200
3
4
5
7 89
0
11
12
4
(2) AC(3) FL (4) PHE(5) ATR
25
50
75
100
125
1
2
6
10 14
13
15
(6) FA(7) PY (8) BaA
min0 5 10 15 20 25 30 350(9) CHR(10) BbFA(11) BkFA (12) BaP
LU : refined pomace olive oil: reference solution (5 µg/kg)(12) BaP
(13) DBahA(14) BghiP (15) IP 600
800
1000
min0 5 10 15 20 25 30 35 400
200
400
23 4 9
10
12
141
56 7 8
11
13
15
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201314
Edible oils: PAH4 Levels
Screening 2012 : 21 vegetable oilsbenzo(a)pyèrenechrysènebenz(a)anthracènebenzo(b)fluoranthène
10
12
EU limit: 10 µg/kg
( )
6
8 PAH4 en µg/kg
2
4
0
ITERG data (2012)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201315
Mineral oilMineral oil
Environmental contamination (air soil)Environmental contamination (air, soil)Crop protectionTransport & storageTransport & storageProduction processR l tiRegulation
(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)
RisksMethodsRegulation RisksMethodsRegulation
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201316
Mineral oil composition
EFSA Scientific Opinion, 2012EFSA Scientific Opinion, 2012
Mineral oil: complex mixture of hydrocarbons
MOSH i h b h d lk & lk l d l lkMOSH: straight or branched alkanes & alkylated cycloalkanes
MOAH: aromatic hydrocarbons including alkyl‐substituted
Hydrocarbon compound number in mineral oil > 100 000 for thosewith less than 20 carbon atoms !
Different products & composition: diesel fuel, white oil, lubricant …
Technical grade mineral oil contain 15‐35 % MOAH, which isminimised in food grade MOSH (white oils)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201317
Mineral oil detected in vegetable oils
2008 contamination of sunflower oil from Ukraine with a mineral oil
Natural hydrocarbonsoil from Ukraine with a mineral oil
from unknown origin
hydrocarbons
2009 contamination of walnut oil with a food grade lubricant oil during refining process
Mineral oil :unresolved complex mixturerefining process
2010 identification of compounds eluted as mineral oil in grapeseed oilseluted as mineral oil in grapeseed oils
2011 contamination of milk fat i h f d d l b i il d i 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14with a food grade lubricant oil during
production
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Contaminated Sunflower oil (MOSH analysis)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201318
MOSH determination: ISO/ CD 17880Fractionation of the sample by liquid chromatography on silica gel or silica gel impregnated with AgNO3
Q ifi i i h i l d d (C18 C20)Quantification with an internal standard (C18 or C20)GC/FID analysis on an short apolar column
Vid
a gel
Vide
1 g oil
5 g Silica
18,
40°C
On‐column injection (2 µl)DB5 HT (15 m‐0,25 mm‐0,1 µm)
Elution with hexane Solvent evaporation GC analysis
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201319
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 chromatogramIt is necessary to integrate twice the chromatogram
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201320
Mineral oil determination:ISO/ CD 17880 pre-collaborative trial ISO/ CD 17880 pre-collaborative trial
450 mg/kg spiked crude soya oil50 mg/kg spiked olive oil
43 participants from 12 countries (2012) dispersed results
550
600
g/kg)
450 mg/kg spiked crude soya oil
140160180
/kg)
50 mg/kg spiked olive oil
450
500
an value
(m
80100120
n value (m
g/
300
350
400
Lab me
0204060
Lab mean
300Laboratories
0Laboratories
sample olive oil crude soya oil
mean value (mg/kg) 62 447mean value (mg/kg) 62 447
reproducibility (mg/kg) 49 153
Horrat value 3,3 1,9
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201321
Phthalates
T t & tTransport & storage
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 foodwith food
Ri kM th dR l ti RisksMethodsRegulation
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201322
Chemical structures of phthalates
Di-methyl PHT DMP R1=R2=methyl
Di th l PHT DEP R1 R2 h lO
Di-ethyl PHT DEP R1=R2=ethyl
Di-isobutyl PHT DIBP R1=R2=isobutyl
Di-butyl PHT DBP R1=R2=butylC
C
O
OR1
Di-hexyl PHT DHexP R1=R2=hexyl
Benzyl butyl PHT BBP R1=benzyl R1=butyl
Di n heptyl PHT DHepP R1 R2 heptyl
C
OR2
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=octylDEHPDi-isononyl PHT DINP R1=R2= isononyl
Di-isodecyl PHT DIDP R1=R2=isodecyl
Oily visquous liquidMW: 390,6 g/molBP: 385°CBP: 385 CWater solubility: 3 µg/lHigh affinity for fat (log Kow : 7,5)
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201323
Phthalates are everywhere
Car undercoatingShoes‐Boots‐GlovesToys Car undercoatingDashboard‐Door panels‐Safety glass
Out‐door & rainwearChild‐care articles
Flooring Roofing Wall coveringCosmetics : Perfume
Ph ti lFlooring‐Roofing‐Wall coveringAdhesives‐Sealant‐RubberPaints‐Shower curtains
Wires & cables‐Fresheners
Hairspray‐DeodorantSkin emollient‐nail polishfingernail elongators
PharmaceuticalsMedical devices :
Catheters‐Blood bag
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 2013
Wires & cables Fresheners
24
(EC) n°10/2011 - Plastic materials & articles into contact with food
Specific migration T b d
articles into contact with food
Specific migration limit in food To be used as
BBP 30 mg/kgPl ti i i i l t i l t i i f ttPlasticizer in single‐use material containing non‐fatty foods except infant formulaeDINP
∑ = 9 mg/kgDIDP
DEHP 1,5 mg/kg Plasticizer in repeated use materials containing non‐fatty foodsDBP 0,3 mg/kg
Material containing these phthalates cannot be used for oils & fats
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201325
DEHP migration in rapeseed oil(PVC t bi )Chromatogram Plots
7 5
MCounts 10-03-2009-05-37-56-CO raff.SMS TIC Filtered50:249
Rapeseed oil before migrationSPME/GC/MS
ITERG data, 2008ITERG data, 2008
(PVC tubing)
5.0
7.5 SPME/GC/MS
0.0
2.5
MCounts 10 03 2009 08 10 47 CO ff + t SMS TIC Filt d
11.6
08 m
inD
MP
13.2
74 m
inD
EP
16.2
39 m
inD
IBP
17.1
76 m
inD
BP
21.0
28 m
inD
Hex
P
+ 23
.939
min
DE
HP
(DIO
P)
26.0
78 m
inD
NO
P
7.5
MCounts 10-03-2009-08-10-47-CO raff + tuyau.SMS TIC Filtered50:249
23.9
42 m
inD
NH
epP
Rapeseed oil after immersion of PVC tubing
DEHP
2.5
5.0
DEHP
10 15 20 25 30minutes
0.0 11.5
82 m
inD
MP
13.2
72 m
inD
EP
15.0
74 m
inD
BA
+ 17
.173
min
DB
P
21.1
15 m
inD
Hex
P
21.3
20 m
inB
BP 26.0
99 m
inD
NO
P
26.0
02 m
inD
IDP
Oil sample kept in a bottling machine tub 3 825 mg/kg of DEHP !
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201326
p p g g/ g
Phthalates: ITERG’s procedurem/z 111 (adipates) m/z 149 (phthalates)GC/MS
DNPP5 50:500
1 g oil
PDMS DEHPDHexP
DNPPDBP
DEP
DMP3
4
5
HS SPME
1 g oil
A l i
DB5ms (30 m; 0,25 mm; 0,25 µm)
DEP
1
2 DNOP
HS SPME Analysis7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0minutes
0
200kCounts
Advantages of the method :
No contamination, no solvent50
100
150
DINP
DIDPRapid analysis (45 min)
Sensitive LOQ < 0,1 mg/kg (excepted DINP & DIDP)25 26 27 28 29 30minute
s
0
50 DIDP
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201327
DEHP in oils & fats
543
mg/kgmg/kg
3
4
mg/kgmg/kg
2
3
0
1
0
ITERG data, 2009
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201328
Conclusions
• Research of contaminants is part of multiple controls conducted byfat and oil industry to fulfill the EC regulation n°1881/2006.
• In the absence of regulation, the detection of contaminants mustbe addressed in partnership with authorities according to thetoxicity of molecules.
• Risks are rather limited due to the efficient elimination during oil‐refining steps
H i b f d d i h d i• However some contaminants can be formed during the productionprocess of vegetable oils such as esters of 3‐MCPD & esters ofglicydol.g y
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201329
A special thanks to….p
ITERG Analysis Department team
COMPOSITIONAL ANALYSIS OF LIPIDS – Ghent – 20‐21 June 201330