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Introduction
Free 3-MCPD (3-monochloropropane-1,2-diol or 3chloropropane-1,2-diol) is first identified on lipid hydrolyzed
by HCl
Introduction
The critical step of refining may be the deodorization. In this
step mono-and di-glycerides, chloride ion and temperature
led to the formation of 3-MCPD monoesters and 3-MCPD
diesters
It is one of the food safety concerns for oils and fats industry
Hot topic in Europe especially among the refiners of oils and
fats
Issue of concern
Dec 2007 German Risk Assessment Body (BfR) issued evaluation
dietary on 3-MCPD esters
Refined oils - highest amount, hence food products using them will
contain elevated amount 3-MCPD esters
Bleaching
Filter Press
Deodorisation
Refined oil
Phosphoric acid
is added
Phosphoric acid
and gums
Gums/Clays and
phosphoric acid are
removed
O
OH
H+
-H2O
OR1
Acylglycerol
Cl
OR1
Cyclic acyl
oxonium ion
OR1
Cl
3-MCPD esters
R = alkyl
R1 = H or COR
3-MCPD
Summary of method:
AOCS Official Method Cd 29a:13
100 mg oil/fat sample
Conversion of 2- and 3-MCPD esters and 3-MBPD esters into free form in acid methanolic solution
GC-MS Analysis
Agilent GC Configuration
7890B GC
Multimode inlet
5977B GCMSD
Extractor ion source
Triple Axis detector
Monolithic Quartz Quadrapole
Mass Hunter software
Standards
1.
2.
3.
4.
5.
Reagents Preparation
Followed AOCS method- Below Screen shot
Standards Preparation
Followed AOCS method- Below Screen shot
Calib 1
(ul)
Calib 2
(ul)
Final
concentration
as 3-MCPD
(ug)
Calib
ISTD
(ul)
Level 0
50
0.39
Level 1
25
0.03
50
0.39
Level 2
50
0.05
50
0.39
Level 3
100
0.10
50
0.39
Level 4
20
0.21
50
0.39
Level 5
30
0.31
50
0.39
Level 6
70
0.72
50
0.39
Level 7
90
0.93
50
0.39
Method
Column Used- DB-1 , 30m x 0.25, 0.25mm FT capillary column
GC Conditions:
1. Injection Volume 1.0 ul
2. Inlet temperature- 250 deg C
3. Injection Mode- Pulsed splitless
4. Inlet Flow- 0.8 ml/min constant Flow
5. Oven Programming- 80C (1 min), from 80C to 170C at 10C/min, from 170C to 200C at
3C/min, from 200C to 300C at 15C/min, 15 min at 300C
MS Conditions
1. Transfer line temperature: 300C
2. Ion source temperature: 230C
3. Quadrupole temperature: 150C
4. Ionization mode: EI, SIM mode
Method Continued
Parameters for SIM (Selected Ion Monitoring) mode:
1. Phenylboronic derivative of 3-MCPD (m/z) 147 (quantifier ion); 196, 198 (qualifier ions);
2. Phenylboronic derivative of 2-MCPD (m/z) 196 (quantifier ion); 198 (qualifier ion);
3. Phenylboronic derivative of 3-MCPD-d5 (m/z) 150 (quantifier ion for 3-MCPD); 201
(quantifier ion for 2-MCPD); 203 (qualifier ion);
4. Phenylboronic derivative of 3-MBPD (m/z) 147 (quantifier ion); 240 (qualifier ion);
5. Phenylboronic derivative of 3-MBPD-d5 (m/z) 150 (quantifier ion); 245 (qualifier ion).
4.4
4.3
4.2
4.1
4
3.9
3.8
3.7
3.6
3.5
3.4
m/z 147
3.3
3.2
3.1
3
2.9
2.8
2.7
2.6
2.5
2.4
2.3
2.2
2.1
2
1.9
1.8
1.7
1.6
1.5
1.4
1.3
1.2
1.1
1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
10.3
10.4
10.5
10.6
10.7
10.8
10.9
11
11.1
11.2
11.3
11.4
11.5
11.6
11.7
11.8
11.9
12
Chromatogram of 2-MCPD
Overlay of Cal 0, Cal 1, Cal 2, Cal 3, Cal 4, Cal 5, Cal 6, Cal 7 & Cal 8 of AOCS Method Cd 29a-13
x10 5
4.5
4.4
4.3
4.2
4.1
4
3.9
3.8
3.7
3.6
3.5
3.4
3.3
3.2
3.1
3
2.9
2.8
2.7
2.6
2.5
2.4
2.3
2.2
2.1
2
1.9
1.8
1.7
1.6
1.5
1.4
1.3
1.2
1.1
1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
m/z 196
m/z 196
phenylboronic derivative of 3-MCPD
10.3
10.4
10.5
10.6
10.7
10.8
10.9
11
11.1
11.2
Counts vs. Acquisition Time (min)
11.3
11.4
11.5
11.6
11.7
11.8
11.9
12
Chromatogram
x10
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
x10
8
7
6
5
4
3
2
1
0
x10
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
10.2
10.3
10.4
10.5
10.6
10.7
10.8
10.9
11
11.1
11.2
11.3
11.4
11.5
11.6
11.7
11.8
11.9
Relative Response
Glycidyl Esters
Gly - 9 Levels, 9 Levels Used, 9 Points, 9 Points Used, 0 QCs
5
y = 1.390515 * x + 0.010148
R^2 = 0.99884432
Type:Linear, Origin:Ignore, Weight:None
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
-0.2
0.2
0.4
0.6
0.8
1.2
1.4
1.6
1.8
2.2
2.4
2.6
2.8
3.2
3.4
3.6
Relative Concentra
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