State of the Art Automated Sample Preparation for 3-MCPD ......3-MCPD - Summary „Sample Prep...

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?

How to get in Contact?

Sales@gerstel.com