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Tackling thechallenge in foodsafety analysis
Dian Maulana
Sales Engineer
PT. Berca Niaga Medika
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GC Overview
Agilent Technologies
Company ProfileBNM is a member of CCM Group
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GC Overview
Agilent Technologies
Authorized Distributor
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GC Overview
Agilent Technologies
Our Key Products Offering
Liquid Chromatography (HPLC)Gas Chromatography (GC)
Mass Spectrometry (MS)
LC-MS, LC-MS/MS, GC-MS, GC-MS/MS
ICP-MS
Microbial Identification System (MIDI)
Agilent Sample Workbench
UV-Visible Spectroscopy
Columns and Supplies
Training, Application, Consulting, and support Services
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Food Industry Testing Segmentation
ResiduesAdditives,
Natural productsContaminantsOther
Pesticides(Biocides)in fruits and
vegetables
Veterinary drugsin
meat and fish
Hormonesin plants
and meatResiduesin animal
feed
Food and Feed authenticity(origin and adulteration) i.e.
Fish and meat speciation;
Feed speciation
Functional (Health) Foods
(Metabolomics)
Food and feed hygiene
(pathogens)
Food and Feed Quality
testing (incl. OOL)Animal and Plant health
GMO
Food terrorism (incl. OOL)
Food contactmaterials
Trace Metals
Mycotoxins
Food allergens
Acrylamide
Dioxins, PAHs
PFOA and PFOS
Contaminants in
animal feed
Human health impact
of contaminants
(metabolomics)
FlavorsColorants
Vitamins and
minerals
Sugars
Fats (i.e. transfats)
Protein contentNutritional labeling
Food enzymes
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Agilent Provide Total Solution for Food Customers
Residues Additives,
Natural productsContaminants
Other
Private Contract Labs
Government Labs
Academia/UniversityFood industry
SamplePreparation GC & GC/MSGC/QQQ
LC & LC/MS
LC/SQLC/TOF
LC/QQQ
Columns &Suppliers Services
Applications and Collaborations
ICP/MS Bioanalyzer
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The world of mass spectrometry
LC/MS
Polar
Labile
Involatile
GC/MS
Non-polar
Stable
Volatile
ICP/MS
Elemental
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Data
Acquisition
and Analysis
HPLC -
Separation
Ion Source
Sample
Introduction
Mass
Spectrometer
Separation and
Measurement
Separation Ionization Detection
LC + MS = LCMS
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Advantages High SpecificityConfidence in results
Excellent QuantitationWide dynamic range
SensitivityLower detection limits
No derivatizationMinimum sample preparation One instrument suitable for many methods
High throughput: multi-analyte analysis
Food Safety Analysis: Meet regulatory requirements
LC-Triple QuadMethod of Choice for Food Safety Analysis
Reduced
Costs aCollision Cell
MS MS
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6495 Triple Quadrupole LC/MS
The most sensitive triple quadrupole in the market
Greater ion sampling and ion transmission efficiency yields better
assay performance for your most demanding applications
6490 Triple Quadrupole LC/MS
Agilent iFunnel TechnologyAttogram zeptomole Sensitivity!
Performance Ultra-Trace Analysis: Ideal for Biomarker and mostChallenging Bioanalytical/Clinical Assays, Steroids, Hormones,
Complex matrices
6460 Triple Quadrupole LC/MS
Agilent Jet Streamsub femtogram sensitivity!
Routine Trace Analysis: Ideal for Multi-Residue Pesticide Analysis,
Mycotoxins, Veterinary drugs
6420 Triple Quadrupole LC/MS
Lowest cost of ownership
Routine Analysis: Ideal for clinical and drug analysis, environmental
analysis
Agilent 6400 Triple Quadrupole Product LineExcellent Value and Performance
6420
QQQ
6495
QQQ
Performance
646
0
Trip
leQua
6460
QQQ
6490
QQQ
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6495 QQQ - Premium Performance
Improvedsensitivity andlowerLLOQs
Average 3x in S/N specifications and applications
Improvedprecision and excellent accuracy atthe lowest levels3x in IDL specifications
Proven 6 orders of linear dynamic range
Proven robustnessin complex matrix
Food matrix and biological matrix (plasma)
Improved mass range, fastscan speed and MRMacquisition rate
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Proven iFunnel Technology
10/6/2014
Internal Sales Training - The new 6495 QQQ LC/MS
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Agilent Jet Stream Hexabore Capillary Dual Ion Funnel
Thermal gradient focusing
Efficient desolvation
Creates an ion rich zone
Six capillary inlets
Samples x10 times more
ion rich gas
Removes the gas but
captures the ions
Removes neutral noise
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Many Ways to Manipulate S/N
Variat ion in S/N measurements makes direct assessment dif f icul t
Increase signal
Increase the gain
Narrow chromatographic peakwidth
Increase scan averaging
Lower noise Select noise region
Narrow the width of noise region
Adjust baseline
Apply peak smoothing & noisefiltering
Vary noise calculation algorithms:Peak-to-Peak, RMS, and ...
a. Peak-to-peak noise
b. RMS noise
c. Smoothing
d. Baseline noise filtering
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Instrument Detection Limit (IDL) is Defined by Statistics
IDLLCMS= t x SD = t x (%RSD / 100) x amount measured
Based on a wel l -establ ished stat is t ical form ula, fo l lows regulatory guid el ines
The minimum amount of analyte that isdetectable and distinguishable frombackground noise with a confidence level
IDL
Student t value, for
99% confidence level
n 1 degree of freedomt
Relative standard deviation / precisionof peak area at the amount measured
From n replicate injections
%RSD
Limited to 2 5 x times higher than theDetection Limit (DL)
Amountmeasured
Theoretical fitting of %RSD is
based on ion statistics
%RSD increases at lower injected
amount
%RSD vs. Injected Amount
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Why Change from S/N to IDL for QQQ LC/MS?Goal of Quantitative AnalysisReporting Detectable AMOUNT
Sensitivity Performance of QQQ LC/MS Lowest detectable amountDetection Limit (DL)
Lowest quantifiable amountLimit of Quantitation (LLOQ)
Criteria of precision and accuracy are met at LLOQ
Signal to Noise (S/N) NOT AMOUNT Variation associated with S/N (noise) measurement
Fails to estimate the true detection limit and LLOQ, when measuredat a high level of S/N > 50:1
Not a good metric of sensitivity performance
Instrument Detection Limit (IDL) AMOUNT A rigorous metric based on statistical analysis of precision (%RSD)
Measured at an analytical low level close to the detection limit
Accurate assessment of the true detection limit and LLOQ
A practical means of evaluating sensitivity performance
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Sensitivity and PrecisionInstrument Detection Limit
1 fg of Reserpine used to measure IDL (+) 1 fg of Chloramphenicol used to measure IDL (-)
IDL = t x (%RSD / 100) x Amount
= 2.821 x (10.9 / 100) x 1 fg = 0.31 fg
IDL = t x (%RSD / 100) x Amount
= 2.821 x (10.0 / 100) x 1 fg = 0.28 fg
6495 IDL Amount measured Replicates Area %RSD t (99%) IDL
Reserpine (+) 1 fg n = 10 10.9% 2.821 0.31 fgChloramphenicol (-) 1 fg n = 10 10.0% 2.821 0.28 fg
1.4
Averag e of 13x 6495 un its
Area %RSD = 10.9
n = 10 injections
Area %RSD = 10.0
n = 10 injections
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Multiple Reaction Monitoring (MRM)
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LC/MS in food safety analysis
Typical food matrices: Fruit and vegetables
Rice, maize, nuts
Meat, poultry, fish
Main analytes of interest:
Pesticides
Antibiotics Mycotoxins
Method of choice:
LC-Triple quad, MRM mode
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Pesticides screening the state of the artPesticide ranking published by EURL for pesticides
Check your scope ranking for pesticides
Listing includes more than 1100 compounds
- Pesticides currently used or used in the past
- Pesticide metabolites of importance
Ranking of pesticides based on:
Toxicological data (toxicological endpoints and endocrine disruptive activity) Residue situation in crops (reporting from labs in Europe and RASFF notifications)
Agricultural usage (including potential for misuse and persistent pesticides)
Analytical informationCompounds Exclusive method
LCMS amenable 683 212
GCMS amenable 552 156
GCMS and LCMS amenable 381
No good method mentioned* 252
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Agilent 1290 Infinity LC + 6460A Triple Quad with Agilent Jet Stream Technology
~600 Dynamic MRM Transitions
9 replicates analyzed at 200 pg level
Mean area %RSD = 3.2, Mean height %RSD= 3.6
Application: Analysis of 300 Pesticides in 15 min.
The combination of UPLC-QQQ with Dynamic MRM acquisition results in high sensitivity,
quantitative data and very fast analysis times
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Unknown Screeningwhats in the
sample?
Qual/Quan:Confirmation and
Quantitation?
Quantitationhowmuch compound(s) is
in the sample?
Non-Selective Selective
Is it Possible to Simultaneously Obtain Qualitative andQuantitative Results?
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A complete, guaranteed
solution
Validated method from
sample preparation to
sample analysis
Guaranteed LOQ and LOD
Complete SOP
Training (optional)
Also available for GC/MS Also available for other
matrices, example: tobacco
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Target analysis of porcine peptide biomarkers inprocessed food commodities with Agilent 6490 QQQ
Jelly
Marshmallow
Soft Gummy
Capsule
Sample preparationEnzymatic digestion
Agilent 6490 LC/QQQ
Target analysis:
Can we find the porcine
peptide biomarkers in these
food samples?
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Confirmation of the targeted porcine peptides by MS/MS
Porcine Marshmallow
Product ion 1
Product ion 2
Product ion 3
Product ion 4
Product ion 1
Product ion 2
Product ion 3
Product ion 4
Bovine Marshmallow
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C L E A R LY B E T T E R L C - M S S O L U T I O N S
Agilent provides the complete solutions
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Agilents MassHunter Workstation
Instrument Control
Real-time monitoring
Method set-up
Autotune
Qualitative Analysis
Chromatographic resultsSpectral results
Find compounds
Quantitative AnalysisUser filters
Compound results
Calibration curve
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Batch-at-a-GlancePowerful Navigation
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Single Vendor Solutions!
Agilent is a Mass Spec Company A complete portfolio of Analyzers and inlets
Continuous Innovations
Agilent is a Chromatography Company
A complete portfolio of LC components Continuous Innovations
Agilent Solutions with Pesticide /Forensics Application Kits
Customizable to meet YOUR needs
Easy to use but POWERFUL
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Single Vendor Solutions!
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240 IT 7000 TQ
220 IT
5975E SQ 5975T LTM SQ
5975C SQ
7200 Q-TOF
More ChoicesBetter SolutionsAgilent GC/MS & GC/MS/MS
ASMS 2011
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Agilent GC/MS Analyzers
Factory setup and
chemically tested
GC/MS/MS Analyzers
for high-sensitive multi-
residue pesticide analysis
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GC/MSSP-1 Pesticide Analyzer Two Configuration Methods
Based on 7890A/7000B GC/MS/MS system, configured with backflushusing
Capillary Flow Technologies. MRM database is included with the method, which is
set up and chemically testedin the factory.
Two configuration methods are offered to meet customers need:
SP1 7890-0501: Method based uponAgilentsRetention Time Locked (RTL) GC/MS/MS
MRM database (500+ pesticides) set up at Constant Pressuremode with Post-column
backflush. It provides flexibility to add GC detectors and easily scaled for shorter runtime.
SP1 7890-0502: Method based upon USA targeted pesticide listRTLGC/MS/MS MRM
database (200+ pesticides) setup at Constant Flowmode with Mid-column backflush. The
method provides ultimate performance and shortest cycle time with reduced carrier gas
consumption.
New !!!
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Individual
Components
produced
Individual
Components
Tested (OFN)
System
assembled
App. method
developed on
cust. system
System checkout
on App. mix
Run 1stcustomer
sample
customization or
optimization
method
On-going
maintains/service
Factory CE Setup; I&F Optional Consulting Aftermarket
Without Ordering Analyzer
Factory SetupOptional
Consulting AftermarketOrder Analyzer CE specialist
I&F
Application
report
System w.
Application setupApp. optimized
Consumables
SP1: Factory Pre-setup Analyzer
Familiarization
& consulting
Std. HW, SW
What is included with the Analyzer?
http://www.sisweb.com/art/sge/gccol.jpghttp://www.agilent.com/about/newsroom/lsca/imagelibrary/images/lsca_98.jpg8/11/2019 Food safety seminar.ppt
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7200 Series Q-TOF for GC/MS
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7200 Analyzer
Doug King
7000B TQ 6500 Q-TOF
NEW
Optics
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New . . . Yet Totally ProvenDual-stage ion m irror improves
second-order time focusing for
high mass resolution.
Hexapole col l ision cel l accelerates
ion through the cell to enable faster
generation of high-quality MS/MS
spectra without cross-talk
Spl i t- f low turbo differentially
pumps the ion source and
quadrupole analyzer compartments
4GHz ADC electronics enable a high
sampling rate (32 Gbit/s) which improves
the resolution, mass accuracy, and
sensitivity for low-abundance samples.
Dual gain amplifiers simultaneously
process detector signals through both low-
gain and high gain channels, extending the
dynamic range to 105.
Analog -to-digi tal (ADC) Detector:
Unlike time-to-digital (TDC) detectors
which record single ion events,ADC
detection records multiple ion events,
allowing very accurate mass
assignments over a wide mass rangeand dynamic range of concentrations.
Proprietary INVAR fl ight tube
sealed in a vacuum-insulated
shell eliminates thermal mass
drift due to temperature changes
to maintain excellent mass
accuracy, 24/7. Added length
improves mass resolution.
Hot , quar tz monol i th ic quadrupole
analyzer and co l l ision cel l identical
to the 7000 Quadrupole MS/MS
Two 300L/s t urbo s pump thefocusing optics and flight tube
From 3 pg of brochure
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OFN Results
theoretical masss = 271.986677 error = -0.5 ppm
1pg OFN
Theoretical Mass = 271.986677 error = -0.5 ppmResolving Power = 13464
RIS, Tom Doherty
0.25 u
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Resolving Power
Resolving Power, R=Mass/PWR = 614/0.68 = 903
Pw=0.68
Mz=614
SQ
IT
TQ
TOF
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Accurate Mass
Pw=0.68
Mz=614
mz < 1ppm
mz = 0.1/614= 160 ppm scan
SQ
IT
TQ
TOF
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240.1218
240.0785
Flurenol methyl ester
m/z = 240.0781
Mass error = 1.7 ppm
Dimetilan
m/z = 240.1217
Mass error = 0.4 ppm
~13,500 resolution FWHM
High Res TOF to Eliminate Isobaric InterferencesAppears as Single 240 u Mass Peak for Any Unit Mass Resolution MS
43
No Intern al Reference Mass (IRM)
correct ions appl ied
m/z = 0.0433
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Q-TOF MS/MS for Structural ElucidationC16H14O4 (Rings + Double Bonds = 10)
44
(MH)+269.0802
Candidatestructures
m/z
(experimental
)
Formula Error
(ppm)
Score
269.0802 C16H13O
4
2.2 80.7
193.0494 C10H9O4 0.6 96.7
167.0334 C8H7O4 3.0 N/A
166.0259 C8H6O4 0.6 N/A
138.0310 C7H6O3 1.1 98.1
110.0359 C6H6O2 3.0 N/A
95.0127 C5H3O2 0.9 99.5
CH2=CHC6H5
CO
CH3
CO
H
C6H5
CH=CHC6H5
Fragment Sequence from
Multiple MS/MS Experiments
Candidate structure #4 fits the
fragmentation information the best
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Agilents ICP-MS Innovations7900 is a successor to 7700 in Single Quad ICP-MS history
1987 2000 20091994First computer-controlled ICP-MS
PMS series
4500
7500
7700
Enabling high
sensitivity
metal analysis
Enabling routine
robust ICP-MS
analysis
Enabling a new level
of interference
management
World's first benchtop system.
Hyperbolic profile quad, motorized torch XYZ,
cool plasma
Enabling a new
level of ease of use
in ICP-MS
9 orders detector range
Octopole Reaction System
HMI
ISIS-DS
3rdGeneration ORS
MassHunter SW
Enabling a new era
in ICP-MS analysis
8800 ICP-QQQ
2012 2014
7900
Worlds First
ICP-QQQ
ICP-QMS Redefined
NewUHMI
NewPlasma interface
and vacuum design
NewDetector system
NewISIS 3
NewMassHunter 4.1
Slide #46
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Agilent 7900 ICP-QMS
Continuing on the success of the Agilent 7700 series ICP-MS:
SMALL in Size, but BIG on Performance
Slide #47
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3 Key Benefits of Agilent 7900 ICP-MSAdvancing the Analytical Dynamics of Food Analysis
1) Matrix Tolerance 2) Interference Removal 3) Dynamic Range
Ultra High Matrix
Introduction System (UHMI)
Handles tough sample
matrices better than any
other ICP-MS. Highest total
dissolved solids(TDS)
capability up to 25% Reduces sample prep time
and error
Better long-term stability
Octopole Reaction System
(ORS4) He Mode
He Mode effectively removes
common polyatomic
interferences in the samples
Easy to use - He collisionmode only
Effective for a wide range of
sample types
Accurate measurements
Wider Dynamic Range
11 orders dynamic range:
0.1ppt(DL) to 10,000ppm
Simplifies method
development
Easy sample prep.
Improves productivity
Facilitates fast TRA
signals, e.g. SP-ICP-MS
Slide #48
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Faster TRA mode facilitates SP-ICP-MS
Minimum dwell time is shortened from 3 msec. on the 7700 Series to 0.1
msec to allow faster sampling of transient signals.
0.1 msecThis enables themeasurement of the
single NP peak signalarriving on the detector.
30 nm Au Nanoparticle
Slide #49
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Agilent 8800 ICP-QQQ
Best New Spectroscopy Product of 2012
Outstanding New Scientific Instrument Product of 2012
R&D100 2013
ACCSIOutstanding
New Scientific
Instrument Product of
2012IBO award
ASMS 2012
Select Science 2012
AwardBest
Spectroscopy Product
Slide #50
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MS/MS reaction cell having high selectivity
MS/MS reaction cell removes the most challenging
interferences.
Phosphorus can be quantified below ppb DL
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Trace Elemental Constituents in FoodsMethod Automation Function (Agilent 7900)
K. Sakai et al.,, Agilent Appl. Note: 5991-4556EN, 30 May 2014..
Slide #54
Challenge
The adoption of new analytical instrumentation can be challenging for food
testing laboratories that have previously used techniques such as ICP-OES and
AAS. Trace level analysis requires tighter control of general laboratory practice
to ensure that the low detection limits can be achieved routinely. ICP-MS, while
offering high sample throughput and low limits of detection, is generally
considered more difficult to use than the well-established ICP-OES and AAStechniques, with one perceived drawback being the greater complexity of
developing a reliable analytical method for new sample types.
Solution
The Agilent 7900 ICP-MS uses ICP-MS MassHunter software, which includes anew Method Automation function to simplify the process of method
development, making it easier for users of all levels of experience to develop
reliable methods for their particular sample type.
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Applications(AdvancedEmerging Trends)
Slide #55
Hg Speciation in Marine Biological Tissues
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Hg Speciation in Marine Biological Tissues
S. Sannac et al., Benefits of HPLC-ICP-MS Coupling for Hg Speciation in Food, Agilent Handbook
of Speciation, 2ndEd., 5990-9473EN (2012).
Gradient LC (1260) conditions:
Start at 2% MeOH
Reaching 90% MeOH in 1 min
90% MeOH till 3 min Going back at 2% MeOH
7700x ICP-MS conditions:
RF: 1,600 W; 8 mm
sampling depth
CRG: 0.54 L/min; MUG:0.10 L/min
Option Gas: 9% (20% O2in
Ar)
Spray Chamber: -5 oC
As Speciation in Rice
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Current CODEX Regulatory Levels*(Total & Inorganic As)
*ftp://ftp.fao.org/codex/meetings/cccf/cccf5/cf05_10e.pdf, viewed 17thJan 2014
Slide #58
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Speciation of As in RiceSynergy of Separation Science and Elemental
Detection
H.R. Hansen, et al. (2012),Arsenic Speciation in RiceA Routine HPLC-ICP-MSMethod, Agilent Handbook of Speciation, 2ndEd., 5990-9473EN.
Sample Preparation 0.2 g of dried and milled acidic extraction (1% HNO3)
overnight Microwave extraction
Centrifuged
Analysis by LC-ICP-MS using 75As with 115In as ISTD
Calibration from 0.5 to 25 ppb
LODs: 0.1-0.3 ppb.
A S i ti i Ri
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As Speciation in Rice
H.R. Hansen, et al. (2012),Arsenic Speciation in RiceA Routine HPLC-ICP-MSMethod, A ilent Handbook of S eciation, 2ndEd., 5990-9473EN.
Species found
in rice were
DMA(V) and
As(V) Total
inorganic As
is the sum of
peak areas for
As(III) and
As(V)
M h 11 2011
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Slide #61
March 11, 2011
Agilent Hachioji
Fukushima
131I
137Cs
90Sr
129I
Radioactive
Contamination
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Summary
Foods analysis is constantly evolving to keep up with
emerging contaminants in life sciences and chemical
analysis
ICP-QMS and ICP-QQQ are now indispensable tools in
modern food testing laboratories due the analytical
dynamics of this technique (sensitivity and specificity).
Emerging frontiers in food analysis will see the analytical
dynamics ICP-MS being used to address both routine and
advanced food analysis applications
Slide #62
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