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State of the Art Automated Sample Preparation for 3-MCPD, MOSH/MOAH, Glyphosate, PEth, and more… Oliver Lerch, Ph.D. GERSTEL GmbH & Co. KG, Muelheim, Germany
State of the Art Automated Sample Preparation for3-MCPD, MOSH/MOAH, Glyphosate, PEth,
and more…
Oliver Lerch, Ph.D.
GERSTEL GmbH & Co. KG, Muelheim, Germany
GERSTEL History
Founded in 1967 in Muelheim an der Ruhr, Germany (headquarter)
Family owned and operated
Agilent Value Added Reseller (VAR), partnership since 1986
6 subsidiaries, >70 distributors, ~200 employees world-wide
Complete solutions for GC and LC
Sample introduction/thermal desorption
Automated sample preparation
Oliver Lerch, Ph.D.
• At GERSTEL since 2005
• Senior Application Scientist until end of 2019
• Strategic applications
• Presentations on seminars, congresses etc.
• Customer demos & trainings
• Support for R&D
• ...
• Beginning 2020Product Manager Automated Sample Preparation
The Benefits of Automation
Excellent, user independent quality of analytical results
Less tedious and monotonous manual work load in the lab
More time for other important tasks:Data evaluation, method development, quality control etc.
Less exposure to toxic solvents and reagents
MultiPurpose Sampler - MPS
Centrifuge
Weighing Option
Extraction
Derivatization
Heating & Cooling
Solid Phase
Extraction SPE
Dilution Wizard
MultiPosition
Evaporation StationmVAP
quickMIX shakerSID processes
1D/2D barcodes
MPS WorkStation
Filtration
Decapper
MAESTRO Control Software
3-Monochloropropanediol (3-MCPD)and Glycidol in
edible Oils and Fats
Background
3-MCPD, 2-MCPD and glycidol as well as their fatty acid esters are process contaminants in edible oils and fats
Ester cleavage in the digestive tract to form free 3- und 2-MCPD or glycidol respectivelyGlycidol probably carcinogenic for humans3-MCPD carcinogenic in animal testingEU and other countries have established maximum levels for glycidol in fats and oilsMaximum levels for 3-MCPD in preparation
Ester cleavage• Saponification under alkaline conditions
Transformation• Glycidol is converted to 3-MXPD; X=halogen
Removal of matrix• Removal of matrix constituents e.g. fatty acid methyl esters by hexane
extraction
Extraction of analytes• MTBE/ethyl acetate 3/2
Analyte derivatizationEvaporation of extract and reconstitution in isooctaneGC-MS(/MS) analysis
+
“Difference Method”- AOCS Cd 29c-13, DGF C-VI 18 (10), ISO 18363-1
3-MCPD Workstation –Usually integrated with GC-MS(/MS)
5x10
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0.9
0.95
1
1.05
1.1
1.15
1.2
1.25
1.3
1.35
1.4
1.45
1.5
1.55
1.6
1.65
1.7
1.75
1.8
1.85
1.9
1.95
2
2.05
+ EIC(198.0, 242.0) SIM 170207-G0504B-01.D
1 2
Counts vs. Acquisition Time (min)12 12.1 12.2 12.3 12.4 12.5 12.6 12.7 12.8 12.9 13 13.1 13.2 13.3 13.4 13.5 13.6 13.7 13.8 13.9 14 14.1 14.2 14.3 14.4 14.5 14.6 14.7 14.8 14.9 15 15.1 15.2 15.3 15.4 15.5 15.6 15.7 15.8 15.9 16 16.1 16.2 16.3 16.4 16.5 16.6 16.7 16.8 16.9 17 17.1 17.2 17.3
2-M
CP
D
3-M
CP
D
3-M
BP
D
Assay A (3-MCPD + glycidol)
Assay B (3-MCPD)
By adding NaBrinstead of NaClglycidol is transformed to 3-MBPD in assay B
Chromatograms
3-MCPD - Summary
„Sample Prep Solution“: Completely automated sample preparation according to AOCS Cd 29c-13 (DGF C-VI 18 (10), ISO 18363-1), including evaporation step
24 samples per 24 hours for online GC-MS(/MS) coupling
28 samples per 24 hours for workstation
LOQ for single quadrupole MS (Agilent 5977) depending on matrix 100 µg/kg
LOQ for triple quadrupole MS (Agilent 7010 B) 10 µg/kg for palm oil
Automation for AOCS Cd 29b-13, Cd 29a-13 and Zwagerman/Overman method available
Mineral Oil Saturated Hydrocarbons (MOSH)&
Mineral Oil Aromatic Hydrocarbons (MOAH)
in Foodstuff and Cosmetics
MOSH MOAH
Packaging and printing
Transport, processing and storage
Environmental impact
In cosmetics: Mineral oil containing ingredients
What are MOSH/MOAH andwhat is their Origin?
DIN EN 16995:2017-08 – LC/GC Coupling Sample extract in hexane
Normal phase LC on silica column
Separation in MOSH and MOAH fraction
Direct transfer to retention gap (LC-GC coupling)
Solvent removal via early vapor exit (EVE)
Parallel GC-FID determination of MOSH and MOAH
Steuerung Open Lab übersicht
Data processing Software
GERSTEL Enterprise Edition powered by OpenChrom
Dedicated software for processing of MOSH/MOAH chromatograms
Works with all standard chromatography data formats
Convenient automated signal integration
Separate MOSH and MOAH chromatogram view for a sample
Standardized report
Summary
Automated MOSH/MOAH analysis solution
Dedicated data evaluation software
Automated epoxidation and ALOX cleanup available
Correct results achieved for proficiency test samples:Rice, cheese, chocolate, cocoa butter, fat and facial cream
>40 samples processed per 24 h following the standard method
LODs for dry, water- and fat-containing foodstuff < 0.5 mg/kg
LODs for fats and oils < 2 mg/kg or < 0.5 mg/kg after cleanup and epoxidation
Glyphosate and its Metabolite Aminomethylphosphonic Acid (AMPA)
in Foodstuff
Glyphosate
Most widely used herbicide in the worldStill approved in the EUScientific controversy:Are glyphosate residues in foodstuff carcinogenic?Acceptable Daily Intake EU (ADI) 0.5 mg/kgEU Maximum Residue Levels (MRL) 0.05 to 20 mg/kg• Wheat 10 mg/kg• Corn 1 mg/kg• Rice 0,1 mg/kg• Tea 2 mg/kg• Most fruits and vegetables 0.1 mg/kg• Drinking water 0.1 µg/L
Derivatization
Homogenized sample
10 mL H2O 15 min soaking
Extraction 30 mL 0.1 M HCl 30 min
Centrifugation
5 mL aliquot + 5 mL 0.1 M KOH
200 µL extract + 800 µL H2O
Add borate buffer (pH 9)
FMOC-Cl derivatization reagent
Derivatization at 50°C
Add 2% H3PO4
Inject 1 mL into SPExos
Online SPE cleanup – LC-MS/MS
Manual steps MPS automation
GERSTEL SPExos II
Glyphosate
Glyphosate in wheat
0.1 mg/kg (green); 0.01 mg/kg (magenta)
AMPA in wheat
0.1 mg/kg (red); 0.01 mg/kg (green)
Cereals, corn, soy beans LOQ ~ 0.01 mg/kg MRL = 0.1 mg/kg for fruits and vegetables (higher for tea, cereals etc.)
Soil LOQ ~ 0.01 mg/kg
Water LOQ ~ 10.0 ng/L MRL = 100 ng/L
Summary and Outlook
Analysis systems are used in routine analysis in contract labs since years
Method extensions planned by GERSTEL
• Inclusion of further analytes (e.g. glufosinate)
• Automation of initial extraction, if feasible
• Optimization regarding cleanup and chromatography
Determination of Phosphatidylethanol (PEth) from Dried Blood Spots (DBS) employing
the Dried Blood Spot Autosampler (DBS-A)
Dried Blood Spots (DBS)
Dried Blood Spot = dried blood sample on cellulose card
Defined area = defined blood volume
Simple sampling, shipment, storage and handling
HemaXis DB 10 for accurate and precise volume whole blood sampling and secure shipment worldwide
Dried Blood Spot Autosampler (DBS-A)Head for card handling
LC pump
Columnoven
SPExos
DBS
Head for optional liquid injection –or collection in
workstation mode
MS
MPS
Non-destructive hematocrit measurement on DBS cards with
NIRFlex N-500
A
B
C
Three operation modes
A: OnlineFlow through desorption, online SPE cleanup, LC/MS analysis
B: WorkstationFlow through desorption, online SPE cleanup, collection and optional further sample prep steps
C: WorkstationFlow through desorption, collection and optional further sample prep steps
Phosphatidylethanol (PEth)
Alcohol consumption marker
Proving single alcohol intake possible up to 12 days after consumption
Drivers´ aptitude tests, workplace drug testing
DBS sample matrix is ideal since PEth can be formed and/or degraded in liquid whole blood
Complete online (A) and workstation (B) analysis method will be available soon
Chromatogram spiked Blank Blood
100 ng/mL per analyte
PEth 16:0/18:1
PEth 16:0/18:2
...and more ???
...and more!
Analysis for microplasticsPesticides in water according to EU water framework directivePolyfluoroalkyl substances (PFAS) in waterPCBs in waste oilTHC and metabolites in serum, hair and urineOpioids, cocaine and metabolites in body liquidsFatty acid distribution (FAMES)Mycotoxins in food and feedAcrylamide in brewed coffeePhthalates in consumer products
GERSTEL team at analytica 2018
Which application do you wish to automate?
