Improvement of efficiency by automation

Wim.Traag@wur.nlMarcel.debeet@wur.nl

RIKILT

Mission

RIKILT contributes to the safety and health

of the Dutch food supply

Antibiotics

ongoing

Dioxins in chickenMay 1999

Melamine in Chinese (milk) products 2007- 2009

Nicotine in mushrooms

2009

Meltdown Tsjernobyl

April 1986DES in meat

1979

MPA in Dutch pig meat

Summer 2002

Chloorm

equat in

pears

1999

BSENovember 1986

Incidents:

Dioxins in Irish pork

2008 /2009

Dioxins in ………..

1999 - 2009

Dioxins in CPP 1998

Dioxins in milk 1989

© FSAI

How did the dioxins get into the feed?How did the dioxins get into the feed?

Contaminated Contaminated Contaminated Contaminated Contaminated Contaminated Contaminated Contaminated

OilOilOilOilOilOilOilOil

BurnerBurnerBurnerBurnerBakery

Waste

Suspected Contamination source: Transformer Oil like Aroclor 1260

© FSAI

•Nearly € 20 Billion Food Industry

•€ 8.16 Billion in Exports in 2008

•1,800 pig processing workers laid

off by 10th Dec

•10,000 Indirect Jobs Threatened

•Reputation Gone for Years

National economy under threatNational economy under threat……..

Automation of DIOXIN method

� Automation is needed :

� To increase sample throughput from 25 � 100 per week

� To reduce delivery time from 10-15 � 3 - 4 workings days

� To improve quality• Better reproducibility � reduction of measurement uncertainty• Better recovery• Etc

� To reduce costs

Method from 1989 -2005

Sample

Sample extraction

Extract clean�up

GC�HRMS

concentration

concentration

spike with 13C labelled standards←

←

←

add clean�up standard

add Recovery standards

Extraction and Clean-up 1989-2005

Semi automated system; combination of three columns using three different HPLC based systems

GPC� Al2O3 � CarbonSoxhlet

In between evaporation is necessaryIn between evaporation is necessaryIn between evaporation is necessaryIn between evaporation is necessary

Concentration

� First evaporation step using the Rotavapor:

From 100 ml to < 5 ml

Takes 15 min

Concentration

� Second evaporation step using the Pierce heating module:

5 ml � 10 µl in six steps

Takes at least 45 min.

Cost of DIOXIN analysis; approach (1989-2005)

� Serie : n =25 +QC

� Sample prep 3 FTE (MBO)

� Measurement using GC-HRMS; 2 FTE (HBO)

� Supervision; 0,5 FTE (scientist)

Cost of instrument

((price / depletion) + maintenance + C))

--------------------------------------------------------- = € xx per day

Days of use

Cost of instrument

((2*350.000 / 5) + (10% van 700.000) + 25000))

--------------------------------------------------------- = € 1500 per day

160

Cost of DIOXIN analysis 1989-2005

� Cost of instrument = € 7.500,--

� Cost of labour = € 20.750,--

� Cost of chemicals = € 2.500,--

� Cost for sample handling = € 825,--

� Total cost = € 30.325,--

� Per sample € 1.200,--

25 samples

Method from 2005

Sample

Sample extraction

Clean�up

GC�HRMS

concentration

concentration

spike with 13C labelled standards←

←

←

add clean�up standard

add Recovery standards

Main Transducer

Pump

Transducer

G

Extract collector

Pressure Gauge

PLE cell.

Solvents

PRESS. Relief Valve

Inlet Valve

Nitrogen

Output valve

Nitrogen Valve

Calibration Valve

5 ml loop

Main Transducer

Pump

Transducer

G

Extract collector

Pressure Gauge

PLE cell.

Solvents

PRESS. Relief Valve

Inlet Valve

Nitrogen

Output valve

Nitrogen Valve

Calibration Valve

5 ml loop

Automated Extraction

Method from 2005

Sample

Sample extraction

Clean�up

GC�HRMS

concentration

concentration

spike with 13C labelled standards←

←

←

add clean�up standard

add Recovery standards

Clean�up after 2005

� Still a three step procedure, however:

� In�line by coupling a combination of four columns

• Acidified silica column + silica column (capacity 3 gram fat)

• Al2O3 column

• Carbon column

PowerPrep

� Purification of ASE�extract

� Silica – oxidation of fat

� Al2O3 – removal of interferences

� Carbon separation ofa. PBDEs & MO�PCBs & Ind.

PCB’s

b. Dioxins & NO�PCBs

21

12

1

2

2

Pump

P.S.

M2 Step 1: Wet Silica Column

1 2 3 4 5 6

1 2 3 4 5

common

com

M5

Alumina

Carbon

Reverse

Forward

M6com

com

M6

M7com

M7

M8

com

Waste

1. Hexane

2. 2% CH2CL2

3. 50% CH2CL2

4. EtAc/Benz

5. Toluene

6. Sample

1. PCDD

2. PCB

3. Fraction 1

4. Fraction 2

5. Fraction 3

6. Waste6

collectors

Power-Prep System/DioxinPCDDs/ PCDFs

Separation Program

1

M3

Silica

com

com

M421

21

21

21

com

com

com

21

21

Step Flow Volume M1 M2 M3 M4 M5 M6 M7 M81 10 50 0 1 1 2 2 0 0 6

M3

M4

M5

com

Legend:

M2 and M8: 6 Way Valves

M3 - M7: 2 Way Valves

POwerprep method rikilt.ppt

Clean�up after 2005

� Both fractions are automatically unattended concentrated using Turbovap (recovery standard and keeper is added)

� Endpoint = 500 ul

� Concentrated fractions are analysed on two GC�HRMS

� Fraction A 10 =l using LVI (Gerstel CIS 4)

� Fraction B 100 =l using LVI (Gerstel CIS 4)

Cost of DIOXIN analysis automated approach

� Cost of instrument = € 7.500,�� € 9.000,��

� Cost of labour = € 20.750,�� € 17.750,��

� Cost of chemicals = € 2.500,�� € 6.500,��

� Cost for sample handling = € 825,�� € 1.650,��

� Total cost = € 30.325,�� € 34.900,��

� Per sample € 1.200,�� � € 700,��

25 samples *1 50 samples *2

*1 Max capacity

*2 Can be increased to 100

� Future

100ml &nitrogen

On-lineevaporation

20mlsharp pulse

ECF Integrated System

Jef Focant

� LIMS

What is SQL*LIMS?

Laboratory

Tests (to be) performed

Tracking progress of samples from receipt until

tests are completed.

Information Management System

Use it to enter information about

Results.

Samples,

� Data stored in an oracle database

� SQL*LIMS

� SQL*LIMS Client application

� Forms, ExcelResults, Reports

WEB based system

SQL*LIMS WAT IS DAT?

Approving

results

Sample receive

Performing

analyses

Principal

Keten van monster tot resultaat.

Workflow from sample to result.

Validation of the results and samples

Analyst.Stores the final results in SQLLIMS: Validation 1.

Specialist.Checks the final results on task�level according to laboratory�specifications: Validation 2.

Projectmanager.Checks the final results on sample� or submissions�level: according to the law. Validation 3.

Reporting sample(s) result(s)

Keten van monster tot resultaat.

Coupling GC-HRMS with LIMS

Radio activity; Coupling of “Gammaster” with LIMS

Summary / Conclusion

� Automation of method achieved by:

� Automated extraction (ASE /PLE)

� Automated clean-up (Power Prep)

� Automated concentration (Turbovap)

� LVI +ALEX (Gerstel)

� Coupling analytical instrument with LIMS

Summary / Conclusion

� Automation of method results in:

� Higher sample throughput (factor 4) with same

number of staff

� Shorter delivery time

� Better quality

� Reduction of cost

Summary / Conclusion

� Coupling analytical instruments to LIMS results in:

� No or less mistakes

� Shorter reporting time

� Better insight in sample flow

Thank you for your attention

Questions?