APGC XEVO TQ-XS - Waters Corporation Marketing… · 2005: AutoSpec Premier 1988: VG AutoSpec and OPUS Data System ©2019 Waters Corporation COMPANY CONFIDENTIAL 5 Time 5.50 6.00

©2019 Waters Corporation 1COMPANY CONFIDENTIAL

APGC XEVO TQ-XS

A Modern Twist to a Complex Analysis….


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Rhys Jones

Senior Research Scientist at Waters

Rhys has worked for Waters in research and development for 19 years,

with a strong focus on gas chromatography mass spectrometry.

The early part of his career was spent developing and supporting the

AutoSpec magnetic sector instrument.

One of his main responsibilities is now the research, development and

support of the atmospheric pressure gas chromatography (APGC)

chemical ionisation source, which has allowed the full range of mass

spectrometry techniques which Waters provides to be applied to GC

analyses.

Your speaker today is:


Waters has a long history of dioxin analysis

1985: VG 70-250 S/SE

1992: VG AutoSpec Ultima2000: AutoSpec Ultima NT 2005: AutoSpec Premier

1988: VG AutoSpec and OPUS Data System


Time5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00 12.50 13.00

%

0

100

100fg of 2,3,7,8 TCDD on AutoSpec

Signal to Noise = 125:1Peak to Peak (±2 SD) using 10 peak widths of noise


EU legislation was revised in 2014 to permit the use of GC-MS/MS for

confirmatory dioxin analysis in food and feed [Commission Regulation (EU) No 589/2014]

GC-MS/MS Dioxin Analysis

2016: Waters Xevo TQ-XS with APGC v2.0


100fg of 2,3,7,8 TCDD on APGC Xevo TQ-XS

Signal to Noise = 5888:1Peak to Peak (±2 SD) using 10 peak widths of noise

Isotope Ratio = 96.2%Expect Ratio = 96.8% Ratio Error = 0.6%



Signal to Noise = 640:1

Ratio Error = 1.8%




Ratio Error = 1.8%


250ag of 2,3,7,8 TCDD on APGC Xevo TQ-XS


Ratio Error = 0.4%


2,3,7,8 TCDD on APGC Xevo TQ-XS

250ag

100ag

Nonane Blank


2,3,7,8 TCDD Dynamic Range and Linearity

linearity within ±8% over the range of 100ag to

100pg


Wellington Labs TCDD-MXB Standard APGC TQ-XS

2fg 1,3,6,8-TCDD

5fg 1,3,7,9-TCDD

10fg 1,3,7,8-TCDD

25fg 1,4,7,8-TCDD

50fg 1,2,3,4-TCDD

100fg 2,3,7,8-TCDDRxi-5Sil MS 60m x 0.25mm x 0.25µm Column


TCDD-MXB Standard diluted 10:1 APGC TQ-XS

200ag

500ag


TCDD-MXB Standard diluted 10:1 APGC TQ-XS

200ag

500ag


Instrument Detection Limit (IDL) Method

Reproducibility of low concentration standard

Signal to Noise Method

Regression of S/N to 3:1 PtP

Limit of Detection 51 attograms

2,3,7,8 TCDD Limit of Detection on APGC Xevo TQ-XS

Limit of Detection 83 attograms

500ag 2,3,7,8 TCDD injections


Atmospheric Pressure Gas Chromatography is a chemical ionisation technique

Source region is filled with nitrogen

A corona pin is used to generate N2+ ions

N2+ ions transfer the charge to the analytes

What is APGC?


Atmospheric Pressure Gas Chromatography

Heated

Transferline

Source

Enclosure



Ionisation

Chamber




Atmospheric Pressure Gas Chromatography G

as V

elo

city (

m.s

-1)

Heated Transfer

Line

Ionisation Chamber

Sample Cone

Source Enclosure

Corona Pin


Atmospheric Pressure Gas Chromatography G

as V

elo

city (

m.s

-1)



Sampling Cone

Electrons

Corona Pin

+2kV



Sampling Cone

Direct Ionisation Region

Corona Pin

+2kV



Sampling Cone

N2+· Nitrogen Ions

Corona Pin

+2kV



Sampling Cone

Corona Pin

+2kV

Neutral Analytes from GC



Sampling Cone

Corona Pin

+2kV

N2+·

N2e-

2e-M+·

M


Comparing High Resolution MS and MS/MS for Dioxins Analysis


MS/MSHigh Resolution MS

Specificity is achieved through high mass

resolution to exclude interferences

Comparing High Resolution MS and MS/MS

Trace analysis in complex matrices requires Specificity

High resolution chromatography provides temporal separation


Nominally isobaric chemical interferences at m/z 322






Specificity is achieved through selection of

characteristic fragmentation




Daughter Ion Scan of 13C12 2,3,7,8 TCDD

-Cl-13CO

Common Loss is COCl


Daughter Ion Scans of 13C12 Dioxin Compounds

TCDD

PeCDD

HxCDD

HpCDD

OCDD

Δm = 13CO35Cl = 63.967 Da


Daughter Ion Scans of 13C12 Furan Compounds

TCDF

PeCDF

HxCDF

HpCDF

OCDF

Δm = 13CO35Cl = 63.967 Da





– Typically requires frequent instrument tuning and

calibrations are essential


Specificity is achieved through selection of

characteristic fragmentation

– Options are limited by chemistry




Hexa Furan

373.821 Da

±0.5 Da

13C12 Hexa Furan

385.861 Da

±0.05 Da

Low Resolution MS – Hexa Furans


High Resolution MS – Hexa Furans

Hexa Furan

373.821 Da

±0.05 Da

13C12 Hexa Furan

385.861 Da

±0.05 Da


MS/MS Data for Hexa Furans

Hexa Furan

373.82 > 310.86

13C12 Hexa Furan

385.86 > 321.89


Tetra Furan Comparison

MS/MS

HRMS


Penta Furan Comparison

MS/MS

HRMS


Hexa Dioxin Comparison

MS/MS

HRMS


Hepta Furan Comparison

MS/MS

HRMS


Isotope Ratios

Theoretical Isotope Profile

TCDD HRMS Data

Relative Abundance = 78.1%


Isotope Ratios in MS/MS

Quadrupole 1 Quadrupole 2Collision Cell

Δm = CO35Cl = 62.96 Da







1 in 4 Probability


Isotope ratios are calculated from the parent ion abundances (as for HRMS)

multiplied by the probability of specified neutral loss

For tetra dioxins:

– Formula Relative Abundance Fragment Probability Combined Probability

– M: C12O2H435Cl4 0.78 1.00 0.78

– M+2: C12O2H435Cl3

37Cl 1.00 0.75 0.75

– Ratio = 0.75 / 0.78 = 0.96




Parent Daughter Parent Daughter

TCDD 319.90 256.93 321.89 258.93 0.96813C12 TCDD 331.94 267.97 333.93 269.97 0.960

TCDF 303.90 240.94 305.90 242.93 0.96813C12 TCDF 315.94 251.97 317.94 253.97 0.960

PeCDD 355.85 292.89 353.86 290.89 0.77613C12 PeCDD 367.89 303.93 365.90 301.93 0.781

PeCDF 339.86 276.90 337.86 274.90 0.77613C12 PeCDF 351.90 287.93 349.90 285.94 0.781

HxCDD 389.82 326.85 391.81 328.85 0.64413C12 HxCDD 401.86 337.89 403.85 339.89 0.640

HxCDF 373.82 310.86 375.82 312.85 0.64413C12 HxCDF 385.86 321.89 387.86 323.89 0.640

HpCDD 423.78 360.81 425.77 362.81 0.80313C12 HpCDD 435.82 371.85 437.81 373.85 0.800

HpCDF 407.78 344.82 409.78 346.82 0.80313C12 HpCDF 419.82 355.85 421.82 357.85 0.800

OCDD 457.74 394.77 459.73 396.77 0.96313C12 OCDD 469.78 405.81 471.78 407.81 0.960

OCDF 441.74 378.78 443.74 380.78 0.96313C12 OCDF 453.78 389.82 455.78 391.81 0.960

AnalytePrimary MRM Secondary MRM

Ratio


APGC MS/MS, EI HRMS and EI MS/MS

Performance Comparison


Time11.50 11.60 11.70 11.80 11.90 12.00

%

0

100

2,3,7,8 TCDD sensitivity EI HRMS vs APGC MS/MS

APGC MS/MS100fg material injected

63,500 ions detected

Electron Ionisation HRMS100fg material injected

9900 ions detected

System “efficiency” – i.e. ions detected vs molecules injected:

0.018% 0.117%


APGC MS/MSElectron Ionisation High Resolution MS

Ions discarded by beam shaping and slitting to

achieve high resolution: 92%

Energy spread rejection: 2%

Beam height restriction: 5%

Final shaping of beam: 10%

Total Losses: 94% (Transmission: 6%)

Typical Ion Losses EI HRMS vs APGC MS/MS

Losses through source ion guide (StepWave),

going from atmosphere to high vacuum: 60%

Quadrupole transmission losses: 15% for each

Collision cell transmission losses: 5%

MRM fragmentation losses (for TCDD): 52%

Total Losses: 87% (Transmission: 13%)


HRMS APGC MS/MS

Analyser Ion Loss 94% 87%

System Efficiency 0.018% 0.117%

Ionisation Efficiency 0.30% 0.90%

So APGC source is producing around three times more ions for TCDD

A further ~ x2 sensitivity comes from using MS/MS instead of HRMS



Most of the source sensitivity can be explained by the “softness” of the ionisation


APGC Spectrum

EI Spectrum

2,3,7,8-[37Cl4]-TCDD

Cleanup Standard

M

13C12 MM - Cl

M - COCl

M – (COCl)2


Softness of ionisation – PCB 209 Example

EI Spectrum

APGC Spectrum


Softness of ionisation – BDE 209 Example

EI Spectrum

APGC Spectrum


Isotope Ratio Accuracy

Detector Saturation

Effects

Low Number

of Ions




How does APGC compare to EI on a tandem quadrupole?

Assessed two EI tandem quadrupole instruments, one an older Waters’ design

with same quadrupole assemblies as TQ-XS

Comparisons made using calibration standards and multicomponent TCDD

sample

APGC MS/MS vs EI MS/MS


Wellington Labs TCDD-MXB Standard APGC TQ-XS

2fg 1,3,6,8-TCDD

5fg 1,3,7,9-TCDD

10fg 1,3,7,8-TCDD

25fg 1,4,7,8-TCDD

50fg 1,2,3,4-TCDD

100fg 2,3,7,8-TCDDRxi-5Sil MS 60m x 0.25mm x 0.25µm Column


TCDD-MXD Standard - EI MS/MS

10fg 1,3,6,8-TCDD

25fg 1,3,7,9-TCDD

100fg 1,3,7,8-TCDD

250fg 1,4,7,8-TCDD

500fg 1,2,3,4-TCDD

1pg 2,3,7,8-TCDD100fg TCDD

Giving approximately 90 ions

1/700th of APGC TQ-XS

[1/100th of HRMS]


TCDD-MXD Standard - Other EI MS/MS system




EI MS/MS Sensitivity

Why do we see these differences?

–HRMS EI sources have a very high extraction field, reducing the effect of charge

suppression

–EI beam shape is not well suited for transmission through a quadrupole

–Atmospheric pressure source allows “cooling” of ions to remove excess energy, giving

better focusing in to quadrupole analyser


Xevo TQ-XS – StepWave XS


Xevo TQ-XS – StepWave XS

Ions and Neutral SpeciesNeutrals removed

through pumping

Ions injected into

first quadrupolePotential applied

between guides


APGC Analytical Performance


Calibration standard concentrations ranges (Wellington Labs) in pg/µL:

Calibration Range

CompoundEN-1948 EPA 1613

CSL CS6 CSL CS5

TCDD/F 0.04 320 0.1 200

PeCDD/F & HxCDD/F 0.08 640 0.5 1000

HpCDD/F 0.16 1280 0.5 1000

OCDD/F 0.16 1280 1.0 2000

13C12 TCDD/F 16 16 100 100

13C12 PeCDD/F & HxCDD/F 16 16 100 100

13C12 HpCDD/F 32 32 100 100

13C12 OCDD/F 32 32 200 200


Diluted by factor of 10 (pg/µL):

Calibration Range

CompoundEN-1948 EPA 1613

CSL CS6 CSL CS5

TCDD/F 0.004 32 0.01 20

PeCDD/F & HxCDD/F 0.008 64 0.05 100

HpCDD/F 0.016 128 0.05 100

OCDD/F 0.016 128 0.1 200

13C12 TCDD/F 1.6 1.6 10 10

13C12 PeCDD/F & HxCDD/F 1.6 1.6 10 10

13C12 HpCDD/F 3.2 3.2 10 10

13C12 OCDD/F 3.2 3.2 20 20


Agilent 7890A GC Oven with 7693A Autosampler

GC Column: Restek Rxi-Dioxin 2, 40m x 0.18mm x 0.18µm

Liner: Single Taper Deactivated

Injector: Split/Splitless in Splitless mode, 290°C, purge time 1.8 minutes

Injections: 1.0µL

Oven:

– 130°C for 1.8 minute,

– 40°C/min to 200°C

– 2°C/min to 235°C

– 3°C/min to 290°C, hold for 12.62 minutes

– Total time 52 minutes

Column flow: 1.4mL/min Helium

GC Conditions


2,3,7,8 TCDD Calibration Line


1,2,3,6,7,8 HxCDD Calibration Line


OCDD Calibration Line


2,3,7,8 TCDF Calibration Line


1,2,3,6,7,8 HxCDF Calibration Line


OCDF Calibration Line


Compound r2 CSL Conc Deviation CSL Isotope Error

2,3,7,8 TCDD 0.999932 -2.0 1.9

1,2,3,7,8 PeCDD 0.999911 5.1 4.2

1,2,3,4,7,8 HxCDD 0.999966 -1.9 -0.3

1,2,3,6,7,8 HxCDD 0.999933 3.6 -2.6

1,2,3,7,8,9 HxCDD 0.999936 -2.4 4.3

1,2,3,4,6,7,8 HpCDD 0.999962 0.3 0.0

OCDD 0.999964 -2.8 2.6

2,3,7,8 TCDF 0.999917 0.1 5.3

1,2,3,7,8 PeCDF 0.999354 10.6 -4.4

2,3,4,7,8 PeCDF 0.999710 11.6 1.6

1,2,3,4,7,8 HxCDF 0.999725 11.2 0.1

1,2,3,6,7,8 HxCDF 0.999996 -0.2 -2.9

2,3,4,6,7,8 HxCDF 0.999891 7.7 -4.4

1,2,3,7,8,9 HxCDF 0.999925 0.0 4.5

1,2,3,4,6,7,8 HpCDF 0.999990 2.6 -4.8

1,2,3,4,7,8,9 HpCDF 0.999990 -1.0 0.9

OCDF 0.999995 0.3 -1.3

Calibration Line Summary


“Perfect” Fit!


Pooled Extract Sample





13C12 2,3,7,8 TCDF

6.0fg 2,3,7,8 TCDF


Palm Oil Sample


Palm Oil Sample


Palm Oil Sample


Vegetable Sample


Vegetable Sample


Vegetable Sample

171fg 1,2,3,7,8 PeCDD

13C12 1,2,3,7,8 PeCDD


Fly Ash QC Sample


Fly Ash QC Sample


Summary


APGC coupled to the Xevo TQ-XS is an extremely sensitive dioxin analyser

– Lowest Limit of Detection and Limit of Quantitation

– Good linearity and dynamic range

MS/MS has similar specificity as HRMS and can perform analysis in complex

matrices

Summary


Advantages of APGC Xevo TQ-XS:

– Sensitivity allows dilution of calibration standards

– No need for large volume injections to meet required detection limits; standard 1µL

splitless, or even split injections work

– Infrequent tuning and calibration when compared to HRMS, weekly or monthly

– Source cleaning only required once or twice a year

– Atmospheric source makes column changes simple

– Also allows very high carrier gas flow rates, useful for large bore columns and very high

boiling point compounds

– Flexible system also capable of LC analysis

Summary


Where to find more information…

waters.com/AnalyticalFoodies

Documents

APGC XEVO TQ-XS - Waters Corporation Marketing… · 2005: AutoSpec Premier 1988: VG AutoSpec and OPUS Data System ©2019 Waters Corporation COMPANY CONFIDENTIAL 5 Time 5.50 6.00