Sample Clean-Up with Immunoaffinity Columns and Gel ......GPC ULTRA Automated Sample Clean-up...

Sample Clean-Up with Immunoaffinity Columns and Gel Permeation

Chromatography: A Product OverviewWestern Canada Trace Organic

WorkshopApril 27, 2010

Wendy RasmussenTechnical Sales Manager

[email protected]

Laszlo TormaDirector, Technical Relations

[email protected]

www.pickeringlabs.com

Background

• Specialists in post-column derivatization• Improvements in equipment, changing

methodologies, led to a need to expand existing product line

• LCTech was an obvious choice:– Partnership already strong (13 years)– GPC Cleanup and Mycotoxin product lines enhance

Post-column offerings

• Now offering the complete product line from LCTech, based in Germany


About LCTech

“We develop innovative products & methods for the analysis of food, animal feed & environment with brilliant support.”

1993 – Begins selling Pickering Products in Europe

1997 – Exclusive Representation of Pickering in Europe

2000 – Introduction of first LCTech GPC Instrument

2005 – Introduction of first LCTech Immunoaffinity column for Aflatoxin


Pickering’s Goals

• Introduce LCTech‘s GPC & Mycotoxin products to North America

• Create New Methods for:– Food and Feed– Environmental

• Be a reliable partner for the analytical community

• Customer satisfaction is our highest priority

• Accept no compromise in quality of products and service


Pickering Offerings

• Pickering as a Manufacturer:– Instruments, Chemistry, Support for Post-column

derivatization

• Key Applications:– Carbamate Pesticides– Glyphosate Herbicide– Amino Acid Analysis– Mycotoxin Analysis– Paralytic Shellfish Toxins– Biogenic Amines


Post-Column DerivatizationInstruments

•Versatile• Improved sensitivity•Reduced reagent use•Reduced maintenance•Automated

•Basic Operation•Cost effective•Simple instrument for basic testing needs

Pinnacle PCX

Vector PCX


30 min. Sodium Amino Acids

1. Aspartic Acid2. Threonine3. Serine4. Glutamic Acid5. Proline6. Glycine7. Alanine8. Cystine9. Valine10. Methionine11. Isoleucine12. Leucine13. Tyrosine14. Phenylalanine15. Lysine16. Ammonia17. Histidine18. Tryptophan19. Arginine


GARD Column Protection System

• No additional back-pressure• No band-spreading• Recommend replacement after 3

months or 300 injections• For Cation Exchange Applications

Carbamate Pesticides

1 Aldicarb sulfoxide2 Butoxycarboxim3 Aldicarb sulfone5 Methomyl6 Ethiofencarb sulfone7 Ethidimuron

8 Thiofanox sulfoxide9 Thiofanox sulfone15 Aldicarb20 Propoxur21 Carbofuran22 Carbaryl

26 Isoprocarb (MIPC)30 Methiocarb31 BDMC32 Bufencarb33 Promecarb35 Aminocarb

Glyphosate Herbicide


Further Offerings

• Pickering as a Distributor:– GPC Clean-up Products

– Immunoaffinity clean-up columns for Mycotoxin– AcceCLEAN Automated column handling

instrument– Elisa Kits

• Combining Pickering with LCTech:– Post-column derivatization– An entire range of products for Mycotoxins –

sample cleanup, analysis


Mycotoxin Product Range

Immunoaffinity Columns

Pinnacle PCXUVE Photochemical Reactor

Elisa Kits

AcceClean Automated Handling System

Mycotoxin Products


IAC Columns

Features:•Diverse Range of Matrices

•Matrix Insensitive

•Easy extraction procedures

•Store at Room Temperature

•Long Shelf-Life: 18months

Typical Matrices:• Grains: corn, wheat, rice, DDG

• Animal Feed

• Nuts: peanuts, pistachio

• Fruits: figs, dates

• Spices: pepper, chili

• Herbs: ginseng, licorice


AcceClean

Valve for up to 7 solvents and air

Three simultaneously operating probes

10 racks, 3 samples each Robust construction

forevery day use

AcceClean Features

Automated Handling system:• 3-columns at the same time• Up to 30 columns per batch• Can be used for IAC, or SPE• Caps remain on column –no draining until proper

time• Up to 7 solvents• 1 port for air to dry• “Soothing Sound” * (virtually silent operation)

*Per Sareeta Nerkar, Pickering R&D Chemist

Recoveries of aflatoxins in peanut butter with 3 mL AflaCLEANTM columns

AcceClean


AcceClean


Column Type MatrixManual

Processing[%]

AcceCLEANTM

Processing [%]

OtaCLEANTM Peanut butter 97 97

OtaCLEANTM Maize 94 93

Afla-OtaCLEANTM Rice 91 93

Post-Column Multi-Residue Mycotoxin Method

• 5 Mycotoxins in a single run• Award winning Single-Laboratory Validated

Method for corn• Can be used for other matrices• Equipment:

– HPLC– Pinnacle post-column derivatization

(Fumonisins)– UVE Photochemical derivatization

(Aflatoxins)– FLD Detection

Multi-Residue Mycotoxin Method

Mycotoxin Standard

Corn Sample


UVE™ Photochemical Reactor

• Derivatization of Aflatoxins• 254 nm UV lamp• High light transmission• Automatic Safety Shut-off• Easy to install & use

0,0 2,5 5,0 7,5 10,0

100

200

300

400mV

min

0

G2

G1

B2

B1


Further Offerings

• Pickering as a Distributor:– GPC Clean-up Products

– Instruments (fully automated to manual), columns

– Immunoaffinity clean-up columns for Mycotoxin– AcceCLEAN Automated column handling

instrument– Elisa Kits

• Combining Pickering with LCTech:– Post-column derivatization– An entire range of products for Mycotoxins –

sample cleanup, analysis


GPC Overview

• Separation according to molecular size• Large molecules (Ex: proteins, lipids, resins) are

eluted from the GPC column while the analytes of interest are retained

• Analytes are eluted with appropriate solvent then concentrated prior to analysis

• The result of GPC clean-up is a very clean sample free from matrix interferences

• Can introduce resulting sample to GC-MS, GC-ECD, HPLC-MS etc.


Flow

High Molecular Compounds –Waste

Analytes -Collect

ForerunMain run

t t

Bio-Beads

Gel Permeation Chromatography


Common applications for GPC

Common Analytes• PCBs• PAHs• Phthalates

(Plasticizers)• Pesticides• Azo Dyes• Dioxins• And Many More....

Common Matrices• Fish• Environmental Samples• Foodstuffs• Spices• Peppermint• Tea• Wheat• Lettuce


Examples of GPC Products

GPC Products

GPC Columns for Pesticides, PCB, PAH, Plasticizers, Mycotoxins, and More

GPC ULTRA

Automated Sample Clean-up Systems for Online-GPC and Concentration

GPC quattro4 Columns parallel

GPC VARIOAutomated GPC System

Note: Photos not shown to scale


Key Features of GPC Ultra

• Silent vacuum pump• Reproducible Concentration & Recoveries• Special septa in sample vials prevent

unwanted solvent evaporation• Specially designed Evaporation chamber• HPLC Pumps designed for long life


Needle Loop

Syringe Pump

Rack

Keypad/Display

Vacuum ChamberAutomatic Drain ValveChiller not shown

GPC Pump

Vacuum Pump

GPC Column

High-Performance Condensers

Solvent Rack

GPC Ultra Components


Rotating Flask

Heating Bath Upper Laser

Lower Laser

Vacuum Chamber

GPC Columns

•Glass, anodized aluminum housing for strength, durability

•No tools required•Easily adjustable bed length•Easy to re-fill•20 bar (290psi) sustained operation*

•QC Tested for performance


*Typical Operating pressures are ~7bar

min10 15 20 25 30 35

pA

20

30

40

50

60

70

80

90

NPD1 A, (D:\GC_4\DATA\SPH605\009F0901.D)

5.1

00 5

.235

5.3

09 5

.473

6.0

46 6

.296

6.8

23 7

.053

7.8

56

8.9

80 9

.126

11.

695

13.

462

14.

770

15.

176

17.

921

20.

140

21.

095

21.

551 25.

068

30.

093

Pirimiphos-methyl

0.2 ppm Pirimiphos-methyl in Wheat

Pirimiphos-Methyl in Wheat

min5 10 15 20 25 30 35 40 45

pA

1000

2000

3000

4000

5000

6000

7000

NPD1 A, (SPH_833\001F0101.D)

min5 10 15 20 25 30 35 40 45

pA

1000

2000

3000

4000

5000

6000

7000

NPD1 A, (SPH_833\002F0201.D)

Subsequent BlankTea Extract

Caffeines

GC-PND Chromatograms showing no carryover of caffeine into blank

No Carry-Over

Pesticides, 0.5g Chicken Fat

0

20

40

60

80

100

120

HC

B

alph

a-B

HC

Lind

ane

Hep

tach

lor

Ald

rin

Ron

nel (

Fenc

hlor

phos

)

Linu

ron

chlo

rpyr

ifos/

oxyc

hlor

dane

Hep

Ep

B

Hep

Ep

A

tran

s-N

onac

hlor

alph

a-C

hlor

dane

/End

osul

I

Chl

orfe

nvin

fos

Die

ldrin

p,p'

-DD

E

Stiro

fos/

Cap

tan

Endr

in

Endo

sulfa

n II

p,p'

-TD

E/o,

p'-D

DT

p,p'

-DD

T

Car

boph

enot

hion

Endo

sulfa

n Su

lfate

Mire

x

Endr

in K

eton

e

Met

hoxy

chlo

r

Phos

alon

e

Cou

map

hos

S

2,2'

,4,4

, 5,5

'-HB

B

Recoveries and Std. Deviation

Recoveries and Standard Deviations (n=60)


Benefits of the GPC Ultra

• Online, reproducible concentration• Eliminates cross-contamination or carry-over

between samples• Wide variety of samples, methods• No loss of solvent from sample vials due to

evaporation• Saves time in the lab by allowing chemists to

pursue other tasks, 24/7 operation• Samples are ready to go directly to analytical

instrument• Low Maintenance

GPC Quattro

Quattro Features:• High Throughput (4 columns)

• Touchscreen Display• Robust• Easy• Automatic shutdown features

– Waste sensors– High-pressure sensors


4 Parallel Columns

Peppermint on GPC Quattro


Overlay of GC-FPD Chromatogram from 10 consecutive samples spiked with chlorinated pesticides

Data courtesy of PhytoLab GmbH in Germany


Summary of GPC Products

• GPC Ultra: Fully automated, online clean-up and evaporation

• GPC Quattro: Multi-column, easy

• GPC is reproducible, rugged, effective for all types of samples

• Contamination and carryover are prevented• Full range of GPC Clean-up products:

– GPC Ultra, Vario, Eva: Fully automated– GPC Quattro: Cost-effective, multi-column, manual


Recap

• Post-column Derivatization Products– Sensitive, Versatile, cost-effective

• Mycotoxin Analysis Products– Versatile, Complete range: from Clean-up

Columns, to HPLC/PCD, to Elisa Kits

• GPC Products– Automated or manual, efficient, reproducible

The Keys

LCTech’s Sample cleanup tools, with Pickering’s Analytical tools leads to efficiency, accuracy, and great results.

Effective Analytical Method!

LCTech’s Effective Sample Cleanup

Pickering’s Guaranteed Chemistry

+ =


Thank You

Have a wonderful day!

Sample Clean-Up with Immunoaffinity Columns and Gel ......GPC ULTRA Automated Sample Clean-up...

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