Recent Status of EU-regulation on 3-MCPD and Glycidol in Oils/Fats, Infant

Formulae and Analytical Solutions Available

J. Kuhlmann
SGS Germany GmbH
jan.kuhlmann@sgs.com

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 1
 Introduction

Free & bound 2-MCPD, 3-MCPD & glycidol - structures

Free analytes (Fatty acid) Bound analytes
just examples, all fatty acids of an oil/fat might be present

FA

3-MCPD  2-MCPD 3-MCPD

FA
T 220 °C
3-MCPD-1,2-bis-palmitoylester
3-MCPD 2-MCPD
3-mono-chloropropane-1,2-diol 2-mono-chloropropane-1,3-diol
3-Chloropropane-1,2-diol FA
2-Chloropropane-1,3-diol
2-MCPD
FA
3-MCPD-1-palmitoylester
+ H+X-
+/- H+Cl-
Analysis
Glycidol FA

3-MXPD FA: all naturally

2-MCPD-1-oleoyl-3-stearoylester
Glycidol occurring fatty acids

“2- & 3-MCPD esters and glycidyl esters”:

glycidol
(2,3-Epoxi-1-propanol)
complex group of contaminants!
Glycidylpalmitate
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 2
 Introduction

Potential hazards of free & bound 2-MCPD, 3-MCPD & glycidol

3-MCPD 2-MCPD Glycidol

OR, OR`= OH; various fatty acids

Free analytes: Toxicity is related to chlorine or an epoxy group at the molecular backbone.

glycidol: genotoxic carcinogen 2A: probably carcinogenic to humans:

➢ no TDI (Tolerable Daily Intake) applies, ALARA-principle (As-low-as-reasonably achievable)
➢ risk assessment based on MoE (Margin of Exposure)

3-MCPD: non-genotoxic carcinogen 2B: possibly carcinogenic to humans, TDI (Tolerable Daily Intake) applies
2-MCPD: No official classification available

EFSA Panel on Contaminants in the Food Chain (CONTAM) : “Animal studies show extensive
hydrolysis of esterified 3-MCPD and glycidol following oral administration; esterified and free forms were
assumed to contribute equally to internal exposures.”
EFSA = European Food Safety Authority
j.efsa.2016.4426
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 3
 Introduction

Occurrence related to infant formulae

crude oil refining refined oil infant formula

➢ In oils/fats bound MCPD/glycidol origins in major parts from the

application of elevated temperatures during refining. Every fully
refined oil is contaminated! But bound glycidol is easier to mitigate
as bound MCPD.

➢ Consequently all infant formula and related nutritionals

containing refined edible oils and fats are prone to also contain 2-
& 3-MCPD esters and glycidyl esters.

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 4
 Risk Assessment

Glycidol: Risk Assessment by MoE

„Hard“ tox data on glycidol[1] : T25/2.5  T10 = 4060 µg/kg bw d
Organisation
Reference Low health concern
BfR 2009[2], 2016 [3], EFSA 2016 [4] MoE ≥ 10 000 Intake ≤ 0.406 µg/kg bw d
hority
BfR: German Federal Institute for Risk Assessment; EFSA: Europen Food Safety Authority
[1]: NTP (1990) Technical Report Series No 374; [2]: BfR Opinion 007/2009; [3] BfR Opinion 020/2016; [4] EFSA Journal 15(5): 4426

➢ T25: Lowest compound uptake level at which (within theire lifetime) 25 % of the tested animals showed
formation of cancer by contrast with a control group.

➢ MoE (Margin of Exposure):

Combination of tox. data with human exposure in order to derive prioritisation of genotoxic compounds.
MoE shall not be used for quantification of toxicity.

➢ To have a low health concern (MoE ≥ 10 000) human exposure should be 10 000 fold lower than the T10.

However,
However, with
with genotoxic
genotoxic
compounds
compounds it it is
is the
the same
same as
as
with
with sports:
sports:
„low
„low health
health concern“
concern“ does
does not
not
mean „no health concern“.
mean „no health concern“.
ALARA-principle
ALARA-principle
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 5
 Risk Assessment

How to use the MoE ?

➢ A Individual MoE-values might be derived by dividing the T10 by the individual intake.

Example: MoE Jan

Estimated Average
vegetable fat intake glycidol level Fat intake Glycidol intake MoE-Calculation MoEJan Health concern
[g/d] [mg/kg] [g/kg bw d] [µg/kg bw d] T10/intake
25 2 0.343 0.685 4060/0.685 5927 elevated
25 1 0.343 0.343 4060/0.343 11 854 low

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 6
 Regulation

Glycidol: Risk Assessment & Regulation

➢ In practice, risk depends upon food consumption.

➢ But people are different and they

consume differently.

➢ How can we estimate MoE-based

maximum glycidol contents in oils?

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 7
 Regulation

Glycidol: Early Risk Assessment 2007-2009

Vegetable fat consumption MoE =≥10
10 000
Max glycidol
Glycidol intake
intake
Average glycidol
Glycidol maxmax content
content in oil/fat
in oil/fat
[g/d]1) [g/kg bw d]2) [µg/kg bw d]
P75 8 0.13 0.406 3000 µg/kg
Women
Max. 32 0.53 0.406 800 µg/kg
P75 20 0.33 0.406 1200 µg/kg
Men
Max. 80 1.33 0.406 300 µg/kg
Infants Max. 63)
6** 0.406 68 µg/kg
Glycidol max content in infant formula (25 % fat)
Infants Max. 63)
6** 0.406 17 µg/kg
1): According to BfR Opinion 047/2007 citing the RKI nutrition survey 2002 2)@ 60 kg adults 3): According to BfR Opinion 007/2009
Limitations of this approach for adults:
➢ Reliability of data regarding fat consumption?
➢ No differentiation of the contamination levels of all types of oils/fats being part of nutrition.
➢ Very different consumption related to age and individual preferences (e.g. vegetarian nutrition)

In practise, risk assessment was carried out much more comprehensive by EFSA (EFSA
Journal 2016/4426) considering several nutrition surveys and ocurrence data.

The simple approach of risk assessment considering the total intake of vegetable fat does
only work for infants.
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 8
 Glycidol: Regulation 2018

Foodstuffs listed in the Annex to

this Regulation that were lawfully
placed on the market before the
entry into force of this Regulation
may remain on the marktet until
19 September 2018

The regulation on glycidol in oils and fats prevents having products on the market with
levels that might cause elevated health concerns.

MoE based Glycidol max content in infant formula (25 % fat)

17 µg/kg

The maximum levels for infant formula seem to be a compromise between risk assessment,
technical feasibility of glycidol mitigation and capability of analytical methods.

Regulation with lower maximum levels on glycidol might be expected even beyond 2019.
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 9
 Risk Assessment

3-MCPD: Tolerable Daily Intake

➢ For TDI determination different mathematical approaches are available.
➢ In case of 3-MCPD this gave different results:

Tolarable Daily Intake (TDI) 3-MCPD

year Organisation/source Analyte TDI [µg/kg bw d]
1994 SCF http://ec.europa.eu/food/fs/sc/scf/out91_en.pdf Free 3-MCPD 2
2007 BfR BfR opinion 047-2007 Bound 3-MCPD 2
5-2016 EFSA j.efsa.2016.4426 Free & bound 3-MCPD 0.8
11-2016 JECFA JECFA/83/SC Free & bound 3-MCPD 4
11-2017 EFSA j.efsa.2018.5083 Free & bound 3-MCPD 2

„The chrystal ball says: In

the EU the authorities will
Basis for draft
now call on the TDI with the
regulation 2016.
highest prevalence.“

BfR: German Federal Institute for Risk Assessment; SCF: Scientific Committee on Food; JECFA: Joint FAO/WHO Expert Committee on Food Additives

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 10
 Regulation

3-MCPD: Risk assessment & draft regulation

Vegetable fat consumption TDI 0.8 µg/kg bw d TDI 2 µg/kg bw d TDI 4 µg/kg bw d
[g/d]1) [g/kg bw d]2) Average 3-MCPD max content in oil/fat
63) 133 µg/kg 333 µg/kg 667 µg/kg
3-MCPD max content in infant formula as powder (25 % fat)
Infants Max. 63) 33 µg/kg 83 µg/kg 167 µg/kg
3-MCPD max content in infant formula as liquid
63) 4 µg/kg 10 µg/kg 20 µg/kg
1): According to BfR Opinion 047/2007 citing the RKI nutrition survey 2002 2)@ 60 kg adults 3): According to BfR Opinion 007/2009

Total 3-MCPD

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 11
 Regulation

Conclusions on the draft regulation 3-MCPD

➢ If the EC follows the EFSA update 2018/5083, a possible future regulation on 3-MCPD might be based on a TDI
of 2 µg/kg bw d.
❖ The Contam Panel stated that the established TDI of 2 µg/kg bw d is not exceeded in the adult
population.
❖ A slight exceedance of the TDI was observed in the high consumers of the younger age groups and in
particular for the scenario on infants receiving formula only.

➢ Do we expect a future regulation on 3-MCPD?

The chrystal ball says: „In

the EU the authorities will
protect the vulnerables and
cut of the peaks.“

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 12
 Analytical methods

Do recent analytical methods meet (draft) regulation? Part I

Vegetable oils and fats intended for direct human consumption or use as an ingredient in food
Analyte (draft) Max. level [µg/kg] Analytical methods (oils/fats) LOQ [µg/kg] Officially validated
Total 3-MCPD (2000) AOCS Cd29b/c 13 100/150 yes
Glycidol 1000 AOCS Cd29a/b/c 13 190/100/150 yes
Vegetable oils and fats destined for the production of baby food and processed cereal-based food
Glycidol 500 AOCS Cd29a/b/c 13 140/100/150 yes

Evaluation of suitable methods:

1. Do methods have the analytes and matrices in the scope? 
2. Is the limit of quantification sufficient? 
3. Is the method officially validated? 

➢ Validated methods for oils & fats are available according to recent regulation on glycidol and also in
perspective to possible future regulation on 3-MCPD!
AOCS Cd 29a,b,c-13 equivalent to ISO DIS 18363-3,2,1 / DGF C-VI 18 (10) equivalent to AOCS Cd29c-13

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 13
 Analytical Methods
Availability of methods for the analysis of 3-MCPD & glycidol in
infant formulae

Several publications are available! [4-10]

[4] Zelinkova Z, Dolezal M, Velisek J;
Occurrence of 3-chloropropane-1,2-diol fatty acid
esters in infant and baby foods.
Eur Food Res Technol 228, 2009, 571–578
[5] A. Becalski, T. Zhao, S. Feng, B.P.-Y. Lau; A
pilot survey of 2- and 3-monochloro-propanediol
and glycidol fatty acid esters in baby formula on
the Canadian market 2012–2013;
J. Food Compos. Anal., 2015, 44, 111-114
[6] Wöhrlin F, Fry H, Lahrssen-Wiederholt M,
Preiss-Weigert A: Occurrence of fatty acid
esters of 3-MCPD, 2-MCPD and glycidol in
infant formula.
Food Addit. Contam., 2015, 32(11), 1810-1822

[7] Wenzel T, Samaras V, Giri A, Buttinger G, Karasek L, Zelinkova Z; Development and [8] Jedrkiewicz R, Głowacz-Rozynska A, Gromadzka J,
validation of analytical methods for the analysis of 3-MCPD (both in free and ester form) and Kloskowski A, Namiesnik J; Indirect determination of
glycidyl esters in various food matrices and performance of an ad-hoc survey on specific food MCPD fatty acid esters in lipid fractions of commercially
groups in support to a scientific opinion on comprehensive risk assessment on the presence of available infant formulas for the assessment of infants’
3-MCPD and glycidyl esters in food, EFSA supporting publication 2015: EN-779 health risk; Food Anal Methods, 2016, 9, 3460-3469

[9] J. Leigh, S MacMahon; Extraction and Liquid
Chromatography-Tandem Mass Spectrometry
Detection of 3-Monochloro-propanediol Esters
and Glycidyl Esters in Infant Formula, J. Agric.
Food Chem., 2016, 64 (49), 9442–9451
[10] Arisseto AP, Silva CS, Scaranelo GR,
Vicente E; 3-MCPD and glycidyl esters in infant
formulas from the Brazilian market: Occurrence
and risk assessment;
Food Control, 2017, 77, 76-81
[11] J. Kuhlmann; Final scientific report on the
decision support project “Investigation into the
presence of 3-MCPD esters and related
compounds in foods.” 2017;
Ref.: 314-06.01-2815HS002

But none of the cited methods has been developed/validated in purpose to match EU regulation.

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 14
 Analytical methods

Do recent analytical methods meet (draft) regulation? Part II

Infant formula, follow-on formula and foods for special medical purposes intended for infants and young children
(powder)
(draft) Maximum level LOQ1),2)
Analyte Analytical methods Officially validated
[µg/kg] [µg/kg]
Total SGS „5-in-2“lowLOQ 15 No*
(125) (2019: 50)
3-MCPD „Nestle´ 2018-a“ 6.5 No*
SGS „5-in-2“lowLOQ 10 No*
Glycidol 75 2019: 50
„Nestle´ 2018-a“ 4 No*
(liquid)
Total SGS „5-in-2“lowLOQ 3 No*
(15) (2019: 6)
3-MCPD „Nestle´ 2018-a“ 3.2 No*
SGS „5-in-2“lowLOQ 2 No*
Glycidol 10 2019: 6
„Nestle´ 2018-a“ 0.7 No*
1):s/n ≥ 10 in milk
*Method proposed to
Evaluation of suitable methods: powder blank
2): RSDr & RSDR sufficient
become AOAC first
1. Are analytes and matrices in the scope of the method?  @ 1st calibration level
action methods
2. Is the limit of quantification sufficient? 
3. Is the method officially validated? ×

➢ Methods for infant formulae are available in perspective to recent and possible future regulation – but they
are not validated so far!
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 15
 Conclusions

Take-home-messages
1) Risk assessment for glycidol is based on MoE, risk assessment for 3-MCPD is
based
based on
onTDI.
TDI.

2) With genotoxic compounds there is no “no health concern”.

3) In the EU a regulation on glycidyl esters in vegetable oils/fats, infant formula and

related foods is in force. Future maximum levels for infant formulae will become
more strict.

4) A draft regulation on 3-MCPD has been circulated but not set into force as different
TDIs were derived by EFSA and JECFA.

5) Regulation also considered technical feasibility of mitigation and analysis.

6) Regarding case 5) a possible future regulation on 3-MCPD in infant formula might

not be predictable by calculating with a “TDI-factor”.

2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 16
 Thank you for your kind attention!
Dr. Jan Kuhlmann; SGS Germany GmbH
Tel.: +49 (0)40 88 309 423
mobile: +49 (0)172 413 8446
Jan.Kuhlmann@sgs.com
2018 AOCS Annual Meeting & Expo, May 06-09 2018; Minneapolis, Minnesota, USA Jan Kuhlmann / SGS Germany GmbH 17