food safety seminar.ppt

8/11/2019 Food safety seminar.ppt

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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


Authorized Distributor


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GC Overview


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


Agilent iFunnel TechnologyAttogram zeptomole Sensitivity!

Performance Ultra-Trace Analysis: Ideal for Biomarker and mostChallenging Bioanalytical/Clinical Assays, Steroids, Hormones,

Complex matrices


Agilent Jet Streamsub femtogram sensitivity!

Routine Trace Analysis: Ideal for Multi-Residue Pesticide Analysis,

Mycotoxins, Veterinary drugs


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.jpg


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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

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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)

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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
ftp://ftp.fao.org/codex/meetings/cccf/cccf5/cf05_10e.pdfftp://ftp.fao.org/codex/meetings/cccf/cccf5/cf05_10e.pdf


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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

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food safety seminar.ppt

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