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Background
Introduction
The goal: totally automated system
for Aflatoxins (B1, B2, G1, G2) and/or Ochratoxin A
From raw extract to chromatogram
=> in 15 minutes
=> without manual steps
HPLC
Introduction
The Components
quaternary or
isocratic pump
sampler
column
oven
detector
HPLC
UVETM
UV derivatiser
(only for Aflatoxins)
Raw extract
Chromatogram
Instrumentation
Sample preparation
and
Analysis
Any HPLC
FREESTYLE Basic
Pump(s)
SPE module
Autosampler
ThermELUTE module
Column oven
Fluorescence detector
ThermELUTE module
Analysis Instrumentation
Any HPLC possible
tested with DIONEX Ultimate 3000
Pump
Isocratic mode
Autosampler
Column switching
valve
Column Oven
Column(s)
Derivatizer
Fluorescence detector
Standard
Software
Standard
ThermELUTE chromatograms
Dionex Ultimate 3000 details
Pump
LPG 3400a
Sampler
Mobile Phase
Methanol/Water/Acetonitrile, 30/60/15
Flow rate
1.2 mL/min
Column Oven
36 C
Column
Flourescence detector
RF 2000, FLD
Fluorescence wavelength
365 nm excitation
460 nm emission
Derivatizer
LCTech UVE
The goal:
Completely automated system
for Aflatoxin and/or Ochratoxin A analysis
Step 1:
Replace 1 mL or 3 mL immunoaffinity columns
with SMART Immunoaffinity columns
SMART Columns
SMART Columns
Loading capacity
100 ng B1
Loading capacity
100 ng Ochratoxin A
Recovery > 80 %
Recovery > 80 %
SMART Columns
SMART Columns
Red wine
Raisins
Liquorice
etc.
etc.
Beer
The goal:
Completely automated system
for Aflatoxin and/or Ochratoxin A analysis
Step 1:
Replace 1 mL or 3 mL immunoaffinity columns
by SMART Immunoaffinity columns
Step 2:
High end Completely automated solution
with FREESTYLE and ThermELUTE module
ThermELUTE
module
Heating element
Injection valve
Width:110 mm
Injection valve
SMART Column
Gripper
ThermELUTE module
Column is transfered
into heating element
The antigen-antibody
bond is broken by the
heat and the toxins are
eluted with water. No
organic solvent is
required.
Heating element (hidden)
SMART Columns
Original size:
25 mm
Advantages
Elution followed by whole sample large volume injection
Advantages
Elution followed by whole sample large
volume injection
Suitable for very strict detection limits
for baby food
Maximum level according Detection limit with
to EU Regulation
FREESTYLE +
1881/2006 *
ThermELUTE
Aflatoxin B1: 100 ppt
Ochratoxin A: 30 ppt
Advantages
Elution followed by whole sample large volume injection
Sample preparation
with FREESTYLETM
Analysis
with HPLC
Automated
sample injection
1st sample
2nd sample
3rd sample
4rth etc
1st sample
2nd sample
3rd etc
Monday
Thuesday
Monday Sunday
Day and night
Loading capacity:
90 samples
(1 SMART column /
sample
& samples presented
in 16 mL vials)
Quantity of samples
can be increased by
reduction of sample
volume and by choosing
smaller vials.
ThermELUTE chromatograms
Main advantages
Completely automated
No cross contamination
Higher sensitivity (15 20x)
Faster analysis
Meets AOAC requirements
Shown in the chromatograms on the following slides
ThermELUTE chromatograms
Ginger
mV
300
40,0
200
30,0
20,0
100
10,0
1
0,0 2
2
1
-50
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
10,0
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Spices: Paprika
mV
250
125
200
100
150
75
100
50
50
25
0 21
2
1
-50
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
-20
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Spices: Nutmeg
mV
Emissi
Emissi
EM:460 nm
250
150
200
150
100
100
50
50
0 21
1
2
-50
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
-20
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Spices: Coriander
mV
250
80
200
150
60
100
40
50
20
1
2
1
2
-50
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
-10
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Nuts: Hazelnut
200
Black:
ThermELUTE detects even
lowest level
150
100
Red:
Not detected with manual
processing
50
0 2
1
-50
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Nuts: Peanut
Emission
Emission
EM:460 nm
mV
250
200
Black:
ThermELUTE detects even
lowest level
150
100
Red:
Not detected with manual
processing
50
0 1 2
-50
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Less sample needed, thus faster processing
700
mV
Cereals: Corn
Corn, 10 ppb
Black:
ThermELUTE, 2.5 mL
500
Red:
ThermELUTE, 1 mL
375
250
Orange:
Manual processing, 3 mL
125
2
1 3
-100
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Less sample needed, thus faster processing
700
Nuts: Hazelnut
Emission
Emission
Emission
EM:460 nm
Hazelnut, 10 ppb
Black:
ThermELUTE, 2.5 mL
500
375
Red:
ThermELUTE, 1 mL
250
Orange:
Manual processing, 3 mL
125
1 3
2
-100
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Less sample needed, thus faster processing
700
Raisins
Emission
Emission
Emission
EM:460 nm
Raisins 10 ppb
Black:
ThermELUTE, 2.5 mL
500
375
Red:
ThermELUTE, 1 mL
250
Orange:
Manual processing, 3 mL
125
2 3
1
-100
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Less sample needed, thus faster processing
700
Nuts: Pistachio
Emission
Emission
Emission
EM:460 nm
Pistachio, 10 ppb
Black:
ThermELUTE, 2.5 mL
500
375
Red:
ThermELUTE, 1 mL
250
Orange:
Manual processing, 3 mL
125
1
2
-100
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
ThermELUTE chromatograms
Less sample needed, thus faster processing
800
Nuts: Peanut
Emission
Emission
Emission
EM:460 nm
Peanut, 10 ppb
Black:
ThermELUTE, 2.5 mL
600
Red:
ThermELUTE, 1 mL
400
Orange:
Manual processing, 3 mL
200
2 3
1
-100
0,0
min
2,0
4,0
6,0
8,0
10,0
12,0
15,0
Methods - Aflatoxin
Nuts: Peanut
Chromatogram of Peanuts
2 mL washing (3mL/min)
Methods - Aflatoxin
Nuts: Peanut
Chromatogram of Peanuts
2 mL washing (3mL/min)
Methods - Aflatoxin
Nuts: Hazelnut
Chromatogram of Hazelnuts
2 mL washing (3mL/min)
Methods - Aflatoxin
Nuts: Hazelnut
Chromatogram of Hazelnuts
2 mL washing (3mL/min)
Ochratoxin
Coffee
Methods - Ochratoxin
Coffee
Methods - Ochratoxin
Wine
Methods - Ochratoxin
Beer
Chromatogram of Beer
SPE Module
GPC Module
EVAporation
Module
ThermELUTE
Module
www.freestyle-robotic.com
www.chromspec.com
sales@chromspec.com
david.mazawa@pickeringlabs.com
Visit us at www.LCTech.de!