sven poggensee agilent technologies hamburg · 2016-08-30 · jp morgan healthcare conf. 2011...

6400 Triple Quadrupole Produkte

Sven PoggenseeAgilent Technologies

Hamburg

6400 Triple Quadrupole Product LineExcellent Value and Performance -

6410 Triple Quadrupole LC/MS

� Robust, Easy-to-Use

� Lowest cost of ownership

� Automate compound optimization

6430 Triple Quadrupole LC/MS� Fast, robust,

Page 2

� Fast, robust,

� Targeted Protein Quantitation

� Fast polarity switching

6460 Triple Quadrupole LC/MS� Agilent Jet Stream – sub fg sensitivity!

� Perfect for the most demanding applications

� Fast polarity switching

� Largest mass range for a premium Triple Quad

All compatible with HPLC chip MS interface

Agilent 6460 QQQ and 6530 QTOF

Rough Pump

Octopole 1

Turbo 1 Turbo 1 Turbo 1

Quad Mass Filter (Q1)

Collision Cell

Lens 1 and 2


10KV Detector

Turbo 2 2nd Turbo

6460 QQQ6530 QTOF

Ion Pulser

Turbo 2

Octopole 1

DC Quad

Rough Pump

Turbo 1 Turbo 1 Turbo 1


Collision CellLens 1 and 2

Octopole 2

*Patent Pending

New 6490 Triple Quadrupole“The highest sensitivity triple quadrupole on the market”

JP Morgan Healthcare Conf. 2011

Zeptmole Sensitivitymicrodosing

dried blood spotshormone analysispersonal care products in waterpesticides –dilute and shoot

Page 4

Dynamic Range of Six Orders Linearityeliminates serial dilution of samplesanalyze trace level impurities in one run

Triggered MRM (tMRM)library searching – NIST data base fit scores

create your own data base for searches

New 6490 Advances Sensitivity 50 Fold in 4 Years

5000

6000

7000

8000

9000

10000

6490S

NR

1 p

g r

eserp

ine

5

0

1000

2000

3000

4000

2006 2007 2008 2009 2010

64106460S

NR

1 p

g r

eserp

ine

iFunnel Technology Captures 6x More Ions

Agilent Jet Stream

• Thermal confinement of ESI plume

• Efficient desolvation to create gas phase ions

• Creates an ion rich zone

Hexabore Capillary

• Samples 6 times more ion rich gas from the source with 6 capillaries

• Captures the majority of the gas from the source region

Dual Ion Funnel

• Removes the gas but captures the ions

• Removes neutral noise

• Extends turbo pump life

June 6

region

Agilent Jet StreamThermal Gradient Focusing Technology

Dramatic Sensitivity Gains for

Premium TOF, Q-TOF, and Triple Quad

Ions Focused in a Collimated

Thermal Confinement Zone

Improved Ionization Efficiency and

Sampling Sampling

Effective Across a Broad Range of

Analyte Classes, including many

APCI compounds

High

Pressure

Stage 1

Low

Pressure

Stage 2

Line of

Sight

Two Stage Ion Funnel Manages the Gas Load

Stage 1

8-12 Torr

Stage 2

1-3 Torr

Offset ion funnels to prevent neutrals from going straight through to MS

June 8

Ion Funnel Operation

RF DriveRF Voltage focuses the ions to the center.

UnfocusedFocused

DC Voltage accelerates the ions to the exit. DC DriveDC Drive

UnfocusedIons and GasEnter

FocusedIons Out

Ion Funnel Construction

Previously, many metal plates made cleaning

a priority because of the large, active surface area…

The high capacitance also required larger power

supplies to provide RF power

Gold Plated

Rim

June 10

The 6490 design uses printed circuit board technology giving a greatly reduces surface area. This low

capacitance enables the use of small power supplies, and enables fast polarity switching

The 6490 design uses printed circuit board technology giving a greatly reduces surface area. This low

capacitance enables the use of small power supplies, and enables fast polarity switching

Simple and Easy Ion Funnel Cleaning Procedure

• As with all LC/MS systems routine cleaning is necessary periodically.

• The high pressure ion funnel should be cleaned periodically, although this could vary from 3 months to 1 year depending on the quantity and type of sampleson the quantity and type of samples

• The high pressure ion funnel is easily removed

• Clean by sonicating the ion funnel assembly in 100% isopropanol for 15 minutes.

1000

Abundance

20 femtogram verapamil on-column injection

ConstantResponse

6490 Performance: For Over 2,000 SamplesProtein Precipitated Plasma over Four Days with 5% RSDs

June 12

Number of Injections

0

500 1,000 1,500 2,000

500

Abundance

• Typically 1 – 5 pg injected for such studies• Lower amounts on-column to monitor response

• Die Software

Agilent 6490 QQQ

• Beispieldaten:

• Anabole Steroide in Urin und Rindfleisch

• Pestizide

• Photoinitiatoren in Verpackungsmaterial

Sven Poggensee

Agilent Technologies

Hamburg

Steroid hormones

• Steroidhormone werden als wachstumsfördernde Medikamente verwendet, um das Tempo des Gewichtsgewinnes zu steigern und Fütterungseffizienz in Schlachtvieh zu verbessern.

• Estradiol, Progesteron und Testosteron sind natürlich vorkommende Steroidhormone und es wird angenommen, daß Verbraucher wenig Risiko haben, falls Nahrung von behandelten Tieren gegessen wird. (FDA)

• Synthetische Hormone wie trenbolone-Acetat, zeranol, und melengestrol-Acetat werden nicht natürlich produziert. Der Metabolismus erfolgt hier wesentlich langsamer als bei natürlichen Hormonen.

• Es gibt ein Verbot für wachstumsfördernde Hormone in der EU.

• Mitgliedsstaaten der EU sind verpflichtet Steroide und Synthetische Hormone in Lebensmitteln zu analysieren

Sven Poggensee

6490 QQQ Hamburg

Page 14

Anabolika in Urin

Target compounds:

• Chlormadinone acetate

• 16-b-Stanozolol

• a/b-Zearalenol

• Ethinyl estradiol

Injection volume: 10 µL

Column: Eclipse PlusC18, 1.8 um, 3.0 x 50 mm

Pump Flow: 0.5 ml/min

Mobile Phase: A = 0.5mM Ammonium Acetate in Water : Methanol (70:30)B = 0.5mM Ammonium Acetate in Water : Methanol (5:95)

Gradient: Time %B

0 251 253 505 756 75

6.01 25 6.01 25

Page 15

Steroids – Chromatography 200 ng/ml All compounds

5x10

3.5

3.75

4

4.25

4.5

4.75

5

5.25

5.5

5.75

6

6.25

6.5

6.75

7

7.25

-ESI MRM Frag=380.0V CID@** (295.2000 -> 145.0000) Cal_11-r001.d

1 1

0.25

0.5

0.75

1

1.25

1.5

1.75

2

2.25

2.5

2.75

3

3.25

Counts vs. Acquisition Time (min)

0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Page 16

5x10

2.6

2.8

3

3.2

3.4

3.6

3.8

4

4.2

4.4

4.6

4.8

-ESI MRM Frag=380.0V CID@** (319.1000 -> 275.1000) Cal_11-r001.d

1 1

Steroids – Alpha and Beta Zearalenol, example of chromatography200ng/ml

Alpha

Beta

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Page 17

1x10

5.25

5.5

5.75

6

6.25

6.5

6.75

7

7.25

7.5

7.75

-ESI MRM Frag=380.0V CID@** (319.1000 -> 160.0000) Cal_1-1ulr004.d Smooth Noise (PeakToPeak) = 2.66; SNR (3.678min) = 7.6

* 3.67820

1 1

-ESI MRM Frag=380.0V CID@** (319.1000 -> 275.1000) Cal_1-1ulr004.d Smooth

Steroids – αααα and ββββ-Zearalenol 0.05ng/ml (50 fg on column with 1uL inj)

αααα-ZearalenolQual ionS/N = 7.6Peak to peak Based on height

ββββ-ZearalenolQual ionS/N = 8.9Peak to peak Based on height

0.0025 ng/ml urine equivalent

2x10

0.7

0.8

0.9

1

1.1

1.2

-ESI MRM Frag=380.0V CID@** (319.1000 -> 275.1000) Cal_1-1ulr004.d Smooth Noise (PeakToPeak) = 6.34; SNR (3.688min) = 5.7

* 3.68836

1 1


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

αααα-ZearalenolQuant ionS/N = 5.7Peak to peak Based on height

ββββ-ZearalenolQuant ionS/N = 4.4Peak to peak Based on height

Page 18

Steroids Low Matrix Spike – Chromatogram

3x10

1

2

3

4

5

6

7

8

-ESI MRM Frag=380.0V CID@** (319.1000 -> 160.0000) UrineSpike_1-r004.d Noise (PeakToPeak) = 28.60; SNR (3.699min) = 253.2

* 3.6997241

* 3.1423768

1 1

-ESI MRM Frag=380.0V CID@** (319.1000 -> 275.1000) UrineSpike_1-r004.d

ββββ-Zearalenol0.04 ng/ml in urine

αααα-Zearalenol0.1 ng/ml in urine

Preconcentration: 20x

4x10

0.2

0.4

0.6

0.8

1

1.2


* 3.69910619

* 3.1425702

1 1


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Page 19

Steroids – Beta Zearalenol – Quant software Batch at a glance

Page 20

1x10

5

5.2

5.4

5.6

5.8

6

6.2

6.4

-ESI MRM Frag=380.0V CID@** (295.2000 -> 143.0000) Cal_2-r001.d Smooth Noise (PeakToPeak) = 2.96; SNR (3.687min) = 2.3

3.6877

1 1

-ESI MRM Frag=380.0V CID@** (295.2000 -> 145.0000) Cal_2-r001.d Smooth

Steroids – Ethynylestradiol 0.1ng/ml (1 pg on column, 10 uL inj)

0.1 ng/ml standardQual ionS/N = 2.3Peak to peak Based on height

0.05 ng/ml urine equivalent

1x10

5

5.2

5.4

5.6

5.8

6

6.2

6.4

6.6

-ESI MRM Frag=380.0V CID@** (295.2000 -> 145.0000) Cal_2-r001.d Smooth Noise (PeakToPeak) = 2.40; SNR (3.678min) = 5.1

3.67812

1 1


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

0.1 ng/nl standardQuant ionS/N = 5.1Peak to peak Based on height

Page 21

2x10

0.5

1

1.5

2

2.5

3

3.5

4

4.5


* 3.691223

1 1


Steroids Low Matrix Spike – Chromatogram

Ethinyl estradiol0.2 ng/ml in urine


2x10

1

2

3

4

5

6

7

8


* 3.691473

1 1


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Page 22

Steroids High Matrix Spike – Chromatogram

3x10

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8


* 3.6911493

1 1


Ethinyl estradiol1.2 ng/ml in urine


3x10

0.5

1

1.5

2

2.5

3

3.5


* 3.6913045

1 1


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Page 23

Steroids – Ethynylestradiol – Quant software Batch at a glance

Page 24

Steroids – Comparison Agilent QQQ 6460 vs. Agilent QQQ 6490200 pg on column

5x10

1.8

1.9

2

2.1

2.2

2.3

2.4

2.5

2.6

2.7

2.8

2.9

3

3.1

3.2

3.3

+/-ESI TIC MRM CID@** (** -> **) 200ng-r001.d

1 1Agilent QQQ 6490Agilent QQQ 6460

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

1.4

1.5

1.6

1.7

1.8


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Page 25

3x10

1

2

-ESI MRM Frag=380.0V CID@** (295.2000 -> 143.0000) VS_Cal_8-r001.d Noise (PeakToPeak) = 9.06; SNR (3.682min) = 206.0

3.6821866

1 1

3x10

2.5

5


3.6853981

1 1

-ESI MRM Frag=160.0V [email protected] (295.0000 -> 143.1000) 200ng-r001.d

Steroids – Comparison Agilent QQQ 6460 vs. Agilent QQQ 6490200 pg Ethinyl estradiol on column

Agilent QQQ 6490

2x10

0.5

1

-ESI MRM Frag=160.0V [email protected] (295.0000 -> 143.1000) 200ng-r001.d Noise (PeakToPeak) = 2.34; SNR (3.472min) = 23.5

3.47255

1 1

2x10

2


3.473189

1 1


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8

Agilent QQQ 6460

Page 26

5x10

1

2


3.698130431

3.140126832

5x10

1

2


3.700171684

3.141165487

-ESI MRM Frag=162.0V CID@** (319.2000 -> 275.2000) 200ng-r001.d

Steroids – Comparison Agilent QQQ 6460 vs. Agilent QQQ 6490200 pg b/a-Zearalenol on column

AgilentQQQ 6490

4x10

1

2

-ESI MRM Frag=162.0V CID@** (319.2000 -> 275.2000) 200ng-r001.d Noise (PeakToPeak) = 10.42; SNR (3.486min) = 1242.2

3.486129442.911

9412

4x10

0.5

1


3.48663702.910

4755


0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5

AgilentQQQ 6460

Page 27

Anabolika in Urin

Target compounds:

• 17-alpha-nortestosterone (aNT)

• 17-beta-nortestosterone (bNT)

• Beta-boldenone (bBOL)

• Norgestrel (NG)

• Trenbolone acetate (TBA)

• Acetoxyprogesterone (AP)


Column: Waters Acquity BEH, 1.8 µm, 100 x 2.1 mm


Mobile Phase: A = H2O 0.1% Formic AcidB = Acetonitrile 0.1% Formic Acid

Gradient: Time %B

0.0 101.0 306.5 738.0 100

10.0 10

Page 28

3.2. Anabolic Steroids - ResultsThe Signal to Noise for the standard injection was determined and the amount injected which would give a signal to noise of three was calculated.

Noise was calculated peak-to-peak and the noise region is indicated in bold.

aNT: 2.5pg injected - S:N=150.548.8 fg injected for a S:N=3

bNT: 2.5pg injected - S:N= 61.2122.5 fg injected for a S:N=3

bBOL: 2.5pg injected - S:N= 269.727.8 fg injected for a S:N=3

TBA: 5pg injected - S:N= 813.918.4 fg injected for a S:N=3

NG: 12.5pg injected - S:N= 378.299.2 fg injected for a S:N=3

AP: 12.5pg injected - S:N= 430.787.0 fg injected for a S:N=3

Page 29

Herausforderung:

> 860 active compounds (Brit. Crop Protection Council)

• > 100 different compound classes

• different physico-chemical properties

• different hetero atoms (halogens, sulfur, nitrogen ...)

• neutral, acidic, and basic pesticides

Pestizide in Nahrung und Umweltproben

• volatile and non-volatile compounds

• complex sample matrices

Page 30

• Need for screening methods; confirmation and quantitation of positives

only (e.g. EU water framework directive)

• Need for “ready to use” methods of analysis including sample

preparation (e.g. EPA Method 1694: “Pharmaceuticals and Personal

Care Products in Water, Soil, Sediment, and Biosolids by HPLC/MS/MS”)

• Increased number of samples in labs:

• Need for faster analysis methods with less sample prep

Environmental analysis trends

• Need for multi-component analyses

• Need for a data management solution

• Ideal:

• easy sample prep

• safe identification

• screening and quantitation in

one analytical run

6490 QQQ Introduction, Berlin

Page 31

Get started quickly

Column & test mix

LCMS Application Kits

Analysis methods show how to use advanced technologies for fast multi-component screening

Column & test mixApplication Note and Quick start guideCD with method parametersLCMS databases for fast screening:

•Dynamic MRM•Accurate Mass

* Contents may vary based upon the application. Ask your Account Manager about the details for your application of interest.


Page 32

Pesticide MRM Data Base G1733AA


Page 33

DMRM Pesticides and Forensic/Tox Database

Analyze medium level standard mixtures

fixed dwell time for each MRM transition using one time segment

MHW Quantitative Data Analysis gets retention times

Load tabular MRM from Database

Compound names, ISTD (optional)

MRM transitions, fragmentorvoltages, collision energies

• Pesticide database contains MRM transitions for more than 600 pesticides

• Forensic/Tox database contains MRM transitions for more than 130 forensic drugs

Create a Dynamic MRM method

Import the results generated in the custom report into acquisition

method

Specify delta retention time window


Page 34

Comparison of MRM and Dynamic MRM

Time (min) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

Compounds (10/block)

Cycle Time (sec)

MRM

50 80

0.5 0.8 1

Time Segment 1 Time Segment 2 Time Segment 3 Time Segment 4

0.7

100 70

Quantitative analysis of pesticides (dMRM)

Cycle Time (sec)

Max Coincident

Cycle Time (sec) 0.4 0.4 0.4 0.4

20 40 40

Dynamic MRM

0.5 0.8 1

30

0.7

• 2 x shorter cycle times supports narrow chromatographic peaks, more analytes or longer dwell per analyte.


Page 35

Dynamic MRM Simulation

Abundance

>>

>

Time >>>

ConcurrentCompounds


Page 36

� Chromatographic conditionsAgilent 1290 Infinity LC system consisting of:

- binary pump

- wellplate sampler

- column compartment

- diode array detector (not used)

HPLC method

Separation column: ZORBAX Eclipse Plus C-18 RRHD column,150 x 2.1 mm, 1.8 µm @ 30°C

4.) Quantitative analysis of pesticides

150 x 2.1 mm, 1.8 µm @ 30°C

Mobile phase: A: 5 mM ammonium formate

B: methanol + 5 mM ammonium formate

Flow: 0.4 ml/min

Gradient: 0.00 min 10 % B0.50 min 10 % B3.50 min 50 % B

12.00 min 100 % B13.00 min 100 % B13.10 min 10 % B15.00 min 10 % B

Inj.Vol.: 1 or 10 µl


Page 37

� Agilent Jet Stream conditions

Spray chamber conditions:Gas temp.: 200°C

Dry gas: 8 l/min

Nebulizer: 30 psi

Sheath gas temp: 300°C

Sheath gas flow: 11 l/min

Positive NegativeCapVoltage: 4500 V 3000 V


CapVoltage: 4500 V 3000 V

Nozzle voltage 300 V 0 V

• Automatic setup of MRM tables based on selected cycle time, retention times and retention time windows for the individual compounds

• Cycle time 500 ms• Interscan delay 3.5 ms• Total No. of MRMs 484• Maximum No. Of concurrent MRMs 68• Minimum Dwell time 3.85 ms• Maximum Dwell time 246.5 ms


Page 38

� Compounds included in method (242 compounds in total)


Acephate

Acetamiprid

Acibenzolar-S-methyl

Acrinathrin

Aldicarb

Aldicarb sulfone

Aldicarb sulfoxide

Amitraz

Atrazine

Avermectin B1a

Azinphos-methyl

Azoxystrobin

Benalaxyl

Bendiocarb

Benfuracarb

Benthiavalicarb

Cinerin 1

Cinerin 2

Clofentezine

Clothianidin

Crufomate

Cyanofenphos

Cyazofamid

Cyflufenamid

Cyhalothrin

Cymoxanil

Cyproconazole

Cyprodinil

Cyromazine

Demeton-S-methyl

Demeton-S-methyl sulfone

Oxydemethon methyl

EPN

Epoxiconazole

Ethiofencarb

Ethiofencarb sulfone

Ethiofencarb sulfoxide

Ethion

Ethofumesate

Ethoprophos

Etofenprox

Etrimfos

Fenamidone

Fenamiphos

Fenamiphos-Sulfone

Fenamiphos-Sulfoxide

Fenarimol

Fenazaquin

Flutriafol

Fluvalinate

Fonofos

Formothion

Fosthiazate

Furalaxyl

Furathiocarb

Haloxyfop acid

Heptenophos

Hexaconazole

Hexazinone

Hexythiazox

Imazalil

Imidacloprid

Indoxacarb

Iprovalicarb

Methidathion

Methiocarb

Methiocarb sulfone

Methiocarb Sulfoxide

Methomyl

Methoxifenozide

Metolachlor

Metolcarb

Metrafenone

Metribuzin

Mevinphos

Monocrotophos

Myclobutanil

Neo-Quassin

Nitenpyram

Nuarimol

Piperonyl butoxide

Pirimicarb

Pirimiphos-ethyl

Pirimiphos-methyl

Prochloraz

Profenofos

Prometryn

Propachlor

Propamocarb

Propanil

Propargite

Propazine

Propetamphos

Propham

Propiconazole

Propoxur

Rotenone

Simazine

Spinosad A

Spinosad D

Spirodiclofen

Spiromefesin

Spiroxamine

Sulfotep

Tebuconazole

Tebufenozide

Tebufenpyrad

Teflubenzuron

Terbufos

Terbufos sulfone

Terbufos sulfoxide

TetrachlorvinphosBenthiavalicarb

Bifenthrin

Bitertanol

Boscalid

Bromuconazole

Bupirimate

Buprofezin

Butralin

Cadusafos

Carbaryl

Carbendazim

Carbofuran

Carbofuran-3-hydroxy

Carbophenothion

Carbosulfan

Chlorfenvinphos

Chlorotoluron

Chlorpyrifos

Chlorthiophos

Oxydemethon methyl

Dialifos

Diazinon

Diclobutrazol

Dicrotophos

Diethofencarb

Difenoconazol

Diflubenzuron

Dimethoate

Dimethomorph

Dimethylvinphos(E)

Dimoxystrobin

Disulfoton-Sulfone

Disulfoton-Sulfoxide

Ditalimfos

Diuron

Dodine

Edifenphos

Emamectin B1a

Fenazaquin

Fenbuconazol

Fenhexamid

Fenoxycarb

Fenpropimorph

Fenpyroximate

Fenthion

Fenthion-sulfon

Fenthion-Sulfoxide

Fipronil

Fluazifop acid

Fluazifop-P-butyl

Fludioxonil

Flufenacet

Flufenoxuron

Fluopicolide

Fluoxastrobin

Fluquinconazole

Flusilazole

Iprovalicarb

Isazofos

Isoprocarb

Isoproturon

Jasmolin 1

Jasmolin 2

Kresoxim-methyl

Lenacil

Linuron

Lufenuron

Malaoxon

Malathion

Mecarbam

Mepanipyrim

Mepronil

Metalaxyl

Metazachlor

Metconazole

Methamidophos

Nuarimol

Ofurace

Omethoate

Oxadixyl

Oxamyl

Paclobutrazol

Paraoxon-ethyl

Paraoxon-methyl

Penconazol

Pencycuron

Pendimethalin

Phenthoate

Phorate Sulfone

Phorate Sulfoxide

Phosalone

Phosmet

Phosphamidon

Phoxim

Picoxystrobin,

Propoxur

Propyzamide

Proquinazid

Prosulfocarb

Pymetrozine

Pyraclostrobin

Pyrazophos

Pyrethrin 1

Pyrethrin 2

Pyridaben

Pyridaphenthion

Pyrifenox

Pyrimethanil

Pyriproxifen

Quassin

Quinalphos

Quinoxyfen

Resmethrin

Tetrachlorvinphos

Tetraconazole

Tetramethrin

Thiabendazole

Thiacloprid

Thiamethoxam

Thiobencarb

Thiodicarb

Tolylfluanid

Triadimefon

Triadimenol

Triazamate acid

Triazophos

Trichlorfon

Trietazine

Trifloxystrobin

Triticonazole

Zoxamide


Page 39

Dynamic MRM viewer – display of concurrent MRMs

Quantitative analysis of pesticides (dMRM)


Page 40

Quantitative analysis of pesticides

� MRM traces of pesticides in sample EF_Cal 3 (10 ng/ml)

• Final method includes 242 pesticides with 2 MRM transitions each• Variable dwell times, Inter-scan delay 3.5 ms


Page 41

Quantitative analysis of pesticides

� MRM traces of pesticides in sample EF_Cal 3 (10 ng/ml)

5x10

0.4

0.45

0.5

0.55

0.6

0.65

0.7

0.75

0.8

0.85

0.9

0.95

1

1.05

+ESI MRM Frag=130.0V [email protected] (890.6000 -> 567.2000) EF_Cal_3.d

more than 120 MRM

transitions in 1.2 min

0.05

0.1

0.15

0.2

0.25

0.3

0.35


9.6 9.7 9.8 9.9 10 10.1 10.2 10.3 10.4 10.5 10.6 10.7 10.8


Page 42

Compounds at a glance – peak review in MH Quant

Page 43


The issue with the matrix …

-100%

-80%

-60%

-40%

-20%

0%

20%

0 5 10 15 20 25 30 35 40 45

ma

trix

eff

ect

s

retentiontime in minutes

� Dilute !!!

,if sensitivity allows ...‘

Page 44

-100%

-80%

-60%

-40%

-20%

0%

20%

0 5 10 15 20 25 30 35 40 45

ma

trix

eff

ect

s


-100%

-80%

-60%

-40%

-20%

0%

20%

0 5 10 15 20 25 30 35 40 45

ma

trix

eff

ect

s


1 : 10 1 : 100

Data provided by Stefan Kittlaus, LUA Sachsen


Pesticides in water

Target compounds:

• Atrazine

• Atrazine-desethyl

• Chlorotoluron

• Cyanazine

• Diuron

• Hexazinone

• Isoproturon

• Linuron




Mobile Phase: A = water + 5mM ammonium formate +

0.01% formic acid

B = 95% acetonitrile/5% water + 5mMammonium formate + 0.01% formic acid

Gradient: Time %B

0.00 20

4.00 70

4.10 100

4.50 100• Linuron

• Metazachlor

• Methabenzthiazuron

• Metobromuron

• Metolachlor

• Metoxuron

• Monolinuron

• Sebuthylazin

• Simazine

• Terbuthylazin


Page 45

10 fg On Column(1 uL inj. of 10 pg/mL)

Monolinuron

0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4

Counts 2x10

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

1.4

1.5

1.6

215.1 -> 126.0 , 215.1 -> 148.1

Ratio = 35.1 (57.0 %)

Acquisition Time (min)

0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4


0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9

Counts 2x10

0

1

2

3

4

5

6

7

202.1 -> 132.0 , 202.1 -> 124.1

Ratio = 82.1 (106.1 %)

Simazine


Page 46


1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4

Counts 2x10

0.25

0.5

0.75

1

1.25

1.5

1.75

2

2.25

2.5

2.75

3

3.25

3.5

278.1 -> 134.1 , 278.1 -> 210.1

Ratio = 58.4 (97.1 %)

Metazachlor


1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4


2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4

Counts 2x10

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

284.1 -> 252.1 , 284.1 -> 176.1

Ratio = 22.4 (111.6 %)

Metolachlor

Qualifier present

Qualifier not present


Page 47


0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2

Counts 2x10

1

2

3

4

5

6

7

8

188.1 -> 146.0 , 188.1 -> 104.0

Ratio = 29.8 (155.7 %)

Atrazine-desethyl


0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2


1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6

Counts 3x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

216.1 -> 174.1 , 216.1 -> 132.0

Ratio = 25.1 (143.4 %)

Atrazine


Page 48


1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4

Counts 2x10

0

1

2

3

4

5

6

7

259.0 -> 170.0 , 259.0 -> 148.1

Ratio = 57.0 (81.7 %)

Metobromuron


1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4

Counts 2x10

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

259.0 -> 170.0 , 259.0 -> 148.1

Ratio = 55.1 (79.1 %)

10 fg on column


1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4


1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 4.1 4.2 4.3 4.4

Counts 2x10

0.5

1

1.5

2

2.5

3

3.5

4

249.0 -> 160.0 , 249.0 -> 182.0

Ratio = 59.1 (86.2 %)

Linuron


Page 49

Summary of Pesticides Results6490 6460 x-fold improvement

pg on column S/N pg on column S/N

Atrazine-desethyl 0.1 33 1 100 3

Hexazinone 0.01 43 1 350 12

Metoxuron 0.01 42 1 120 35

Simazine 0.1 59 1 75 8

Cyanazine 0.1 23 1 65 4

Methabenzthiazuron 0.01 42 1 125 34

Chlortoluron 0.01 56 1 95 59

Atrazine 0.1 123 1 105 12Atrazine 0.1 123 1 105 12

Isoproturon 0.01 77 1 250 31

Monolinuron 0.01 14 1 80 18

Diuron 0.01 33 1 50 66

Metobromuron 0.1 83 1 30 28

Metazachlor 0.01 43 1 280 15

Sebuthylazin 0.01 29 1 250 12

Terbuthylazin 0.01 38 1 280 14

Linuron 0.1 31 1 20 15

Metolachlor 0.01 65 not recorded


Page 50

Photoinitiators

• More than 1000 compounds are used for printing on packaging materials for food. In 2005 ITX (2-Isopropylthioxanton) a photoinitiator has been found in baby food packed in paperboard containers and had been taken off the shelves in Italy.

S

CH3

CH3

O

• Photoinitiators are compounds which accelerate the drying of colors which undergo a UV induced polymerization.

• ITX is non-mutagenic but it is not acceptable in food products and at concentrations above 0.05 mg/kg food product EFSA requested food producers to react.

• Nestlé has published a list of compounds which are accepted in food packaging, all other compounds need to be tested for food migration and must comply with regulations or must be at concentrations below 0.01 mg/kg.


Page 51

PhotoinitiatorsScreening method

Target compounds:

• 1-(4-((4-Benzoylphenyl)-thio)-phenyl)-2-methyl-2-((4-methylphenyl)-sulfonyl)-1-propan-1-on

• 1-Chlor-4-propoxythioxanthon

• 1-Hydroxycyclohexylphenylketon

• 2-(4-Methylbenzyl)-2-dimethylamino-1-(4-morpholinophenyl)-1-butanon

• 2,2-Bis-(2-chlorophenyl)-4,4,5,5-tetraphenyl-1,2-biimidazol

• 2,2-Diethoxy-acetophenon

• 2,2-Dimethoxy-2-phenylacetophenon

• 2-Hydroxy-4-methoxybenzophenon

• 2-Hydroxy-4-n-octoxy-benzophenon

• 3-Methylbenzophenon

• 4-(4-Methylphenylthio)-benzophenon

• 4,4,4-Methylidynetris-(N,N-dimethylanilin)

• 4,4-Bis-(diethylamino)-benzophenon

• 4,4-Bis-(dimethylamino)-benzophenon

• 4-Dimethylaminobenzophenon

• 4-Hydroxybenzophenon• 2,2-Dimethoxy-2-phenylacetophenon

• 2,4,6-Trimethylbenzophenon

• 2,4,6-Trimethylbenzoylphenylphosphinsäureethylester

• 2,4-Diethylthioxanthon

• 2-Benzyl-2-dimethylamino-4-morpholinobutyrophenon

• 2-Chlorthioxanthon

• 2-Dimethylamino-ethylbenzoat

• 2-Ethylanthraquinon

• 2-Ethylhexyl-4-dimethylaminobenzoat

• 2-Hydroxy-1-(4-(4-(2-hydroxy-2-methylpropanyl)-benzyl)-phenyl-2-methyl-2-propanon

• 4-Hydroxybenzophenon

• 4-Phenoxy-2,2-dichloracetophenon

• 4-Phenylbenzophenon

• Benzophenon

• Bisphenol A

• Butoxyethyl-4-dimethylamino-benzoat

• Diphenyl-2,4,6-trimethylbenzoyl)-phosphinoxid

• Ethyl-4-dimethylamino-benzoat

• Isopropylthioxanthone

• Methyl-2-benzoylbenzoat

• Phenyl-bis-(2,4,6-trimethylbenzoyl)-phosphinoxid

• Thioxanthon


Page 52

Photoinitiators in cardboard extract




Mobile Phase: A = water + 5mM ammonium formateB = acetonitrile

Gradient: Time %B

0 100.5 103.5 609 95

11.5 9511.6 10


Page 53

Photoinitiators – Chromatography 100 ng/ml (all compounds)

6x10

2.2

2.4

2.6

2.8

3

3.2

3.4

3.6

3.8

4

4.2

+ESI TIC MRM (** -> **) Mix_35_100ppb+BP.d

1 1

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2


0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11


Page 54

Photoinitiatoren – Quant software Batch at a glance

2-Hydroxy-1-(4-(4-(2-hydroxy-2-methylpropanyl)-benzyl)-phenyl-2-methyl-2-propanon


Page 55


Methyl-2-benzoylbenzoat


Page 56


1-(4-((4-Benzoylphenyl)-thio)-phenyl)-2-methyl-2-((4-methylphenyl)-sulfonyl)-1-propan-1-on


Page 57

Bisphenol A and benzophenone – Comparison Agilent 6460 vs. 6490100 ng/ml in muesli extract, 5 ul

Agilent QQQ 6490Agilent QQQ 6460


Page 58

Bisphenol A – Comparison Agilent QQQ 6460 vs. 6490100 ng/ml in muesli extract, 5 ul

30 X Signal12 X SNR

Agilent QQQ 6490

Agilent QQQ 6460


Page 59

LOQ 2.5 pg o.c.(S/N 10, Peak-to-Peak)

Benzophenon – Comparison Agilent QQQ 6460 vs. 6490100 ng/ml in muesli extract, 5 ul

9 X Signal6 X SNR

Agilent QQQ 6490

Agilent QQQ 6460


Page 60

LOQ 2.5 pg o.c.(S/N 10, Peak-to-Peak)

New triggered MRM functionality for confirmation


Page 61

Identification of compounds by library search - Qual

Library search functionality in MassHunter Qual

Excellent peak shapes for primary transitions!


Page 62

Identification of compounds by library search - Quant


Page 63

Qualifier ratios in triggered spectra

0.0

2000.0

4000.0

6000.0

8000.0

10000.0

12000.0

14000.0

16000.0

105.1 133.1 139.00 146.00 331.1 360.00

1-(4-((4-Benzoylphenyl)-thio)-phenyl)-2-methyl-2-((4-methylphenyl)-sulfonyl)-1-

propan-1-on

m/zin spectra

ratioRSD

Peak Area ratio

105.1 100.0 0.0

133.1 30.4 18.5

139.0 25.1 19.6

146.0 54.4 4.4

331.1 89.7 5.7 80.6

360.0 47.6 6.8

0.0

10000.0

20000.0

30000.0

40000.0

50000.0

60000.0

127.00 171.00 200.00 228.00 234.00 263.00

1-Chlor-4-propoxythioxanthon

m/zin spectra

ratioRSD

Peak Area ratio

127.0 47.0 17.6

171.0 74.8 0.7 74.9

200.0 100.6 16.4

228.0 175.4 19.7

234.0 9.6 22.8

263.0 100.0 0.0


Page 64

0.0

10000.0

20000.0

30000.0

40000.0

50000.0

60000.0

70000.0

80000.0

90000.0

100000.0

89.10 165.10 191.00 226.00 329.10

2-(2,2-Bis-(2-chlorophenyl)-4,4,5,5-tetraphenyl-1,2-biimidazol

Qualifier ratios in triggered spectra

m/zin spectra

ratioRSD

Peak Area ratio

89.10 21.9 0.86 21.8

165.10 46.1 19.76

191.00 18.3 20.14

226.00 16.5 21.99

329.10 100.0 0.00

0.0

20000.0

40000.0

60000.0

80000.0

100000.0

120000.0

140000.0

77.10 105.10 165.10 197.10

2-(2,2-Dimethoxy-2-phenylacetophenon

m/zin spectra

ratioRSD

Peak Area ratio

77.10 100.0 0.00

105.10 377.8 18.22

165.10 185.5 17.22

197.10 93.0 0.79 93.7


Page 65

5. Summary and Conclusions

• It was demonstrated that the 1290 Infinity UHPLC coupled with a 6490 QQQ gives excellent sensitivity for the three analyte groups.

• The detection limit of almost all analytes was improved over the 6460 QQQ, in many cases by more than a factor of 10.

• Although highly sensitive, the 6490 QQQ performs with great robustness, which shows in the good reproducibility of the peak area.


Page 66

Vielen Dank für Ihre Aufmerksamkeit

Sven PoggenseeAgilent TechnologiesSales & Service GmbH & Co KG

?

Sales & Service GmbH & Co KG

[email protected]

Page 67


sven poggensee agilent technologies hamburg · 2016-08-30 · jp morgan healthcare conf. 2011...

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