Improvement of efficiency by automation
[email protected]@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?