First Impressions on the

Agilent 7200 GC / Q-TOF

Frank David

RIC and Ghent University

Mass Spectroscopy

Structure

Elucidation

Selective and

Sensitive Detection

Identification Target Compound

Analysis

Mass Spectroscopy

Structure

Elucidation

Selective and

Sensitive Detection

Identification

(NIST, Wiley)

Target Compound

Analysis

Mass Spectroscopy

Structure

Elucidation Selective and

Sensitive Detection

Identification: accurate mass

“Open Methods”

Target Compound

Analysis by MS/MS

The (GC) MS compromise triangle

Selectivity

(Resolution) Mass Accuracy

(ID) Robustness

Sensitivity

“Scan” versus “SIM” (MRM)

QQQ

(Q)TOF

SQ (scan)

S/N

n (solutes)

SQ (SIM)

Applications & Expectations

• Analysis of pesticide mixture – Sensitivity check on aldrin (organochlorine pest)

– Searchable spectrum at low pg level?

• Analysis of urinary metabolites in kidney disease study – Compare with MSD

• Analysis of Biomarkers in Crude Oil – Resolution, extract accurate mass chromatograms

– MS/MS operation

Analysis by GC-MSD (5975C) - SCAN 10 pg on-column - EIC (262.8)

Aldrin: 15.60 min

1 2 .0 0 1 3 .0 0 1 4 .0 0 1 5 .0 0 1 6 .0 0 1 7 .0 0 1 8 .0 0 1 9 .0 00

1 0 0

2 0 0

3 0 0

4 0 0

5 0 0

6 0 0

7 0 0

8 0 0

9 0 0

1 0 0 0

1 1 0 0

1 2 0 0

1 3 0 0

1 4 0 0

1 5 0 0

1 6 0 0

1 7 0 0

1 8 0 0

1 9 0 0

2 0 0 0

2 1 0 0

2 2 0 0

2 3 0 0

T im e -->

A b u n d a n c e

Io n 2 6 2 .8 0 (2 6 2 .5 0 to 2 6 3 .5 0 ): s q te s t-1 4 .D \d a ta .m s

1 5 .6 0 2

1 6 .1 3 4

1 6 .3 7 5

1 7 .5 1 0

1 7 .9 0 2

Analysis by GC-MSD (5975C) - SCAN MS (background substrated) – NIST search

Scan 1047 (15.602 min): sqtest-14.D\ data.ms (-1045) 1,4:5,8-Dimethanonaphthalene, 1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-, (1à,4à,4aá,5à,8à,8aá)-Head to Tail MF=449 RMF=523

50 70 90 110 130 150 170 190 210 230 250 270 290 310 330 350 370 390 410 430 450 470 490

0

50

100

50

100

51

64

66

73

79

81

91

91

96 116

121

143

143

159

160186

191

201

207

216

220

235

237

248

255

263

263

293

298 324

329

341 355

364

390428 450462 480492

Hit quality: match = 449; Rmatch = 523; Prob = 28.9

Analysis by GC-QQQ (7000B) - SCAN 10 pg on-column - EIC (262.8)

Aldrin: 17.93 min

Analysis by GC-QQQ (7000B) - SCAN MS (background substrated) – NIST search

+EI Scan (17.934 min) test04.D Subtract 1,4:5,8-Dimethanonaphthalene, 1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-, (1α,4α,4aβ,5α,8α,8aβ)-Head to Tail MF=612 RMF=635

50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450 460

0

50

100

50

100

50

51

55

66

66

75

79

84

85

91

92

97

101

101

110

111

127

133

147

149 158

160

176

186

191203

205

207

220

227

229

241

257

263

265

276

281 293

293

314

315

327

329

341 356

364

366 405 417 452

Hit quality: match = 612; Rmatch = 635; Prob = 67.9

Analysis by GC-QTOF (7200) 10 pg on-column - EIC (262.8564)

Aldrin: 18.53 min S/N = 147

Analysis by GC-QTOF (7200) 10 pg on-column - Spectrum

Analysis by GC-QTOF (7200) 1 pg on-column - EIC (262.8564)

GC-MS based metabolomics

• Agilent Fiehn GC/MS Metabolomics RTL Library (G1676AA)

• A retention time locked (RTL) GC-MS method on a 7890A GC with 5975C inert XL MSD

• Library containing searchable EI spectra and retention times from appx. 700 common derivatized metabolites

• DB5-MS (30 m x 250 µm x 0.25 µm) • 60°C - 1 min - 10°C/min - 325°C • Splitless, 250°C • CF, 1.1 mL/min • MS in scan (50-600 amu)

Data handling: differential analysis Healthy

Disease

Feature extraction

Alignment, normalization and filtering

(MassProfilerPro)

H1

H2

H3

H4

H5

D1

D2

D3

D4

D5

Data handling: differential analysis

Feature extraction

Alignment, normalization and filtering

(MassProfilerPro)

PC1

PC2

PC3

Statistical analysis (MassProfilerPro)

GC-MSD analysis of Urine sample Fiehn method (oximation – silylation)

8 . 0 0 1 0 . 0 0 1 2 . 0 0 1 4 . 0 0 1 6 . 0 0 1 8 . 0 0 2 0 . 0 0 2 2 . 0 0 2 4 . 0 0 2 6 . 0 0

5 0 0 0 0 0

1 0 0 0 0 0 0

1 5 0 0 0 0 0

2 0 0 0 0 0 0

2 5 0 0 0 0 0

3 0 0 0 0 0 0

3 5 0 0 0 0 0

4 0 0 0 0 0 0

4 5 0 0 0 0 0

5 0 0 0 0 0 0

T i m e - - >

A b u n d a n c e

T I C : 1 1 0 4 2 0 _ C K D 3 - 1 6 . D \ d a t a . m s

CKD3

GC-QTOF: diluted 1/10 versus MSD

Healthy versus CKD3

From differential analysis

First hit: Formula (M): C10 H16 O Si Mass (MFG): 180.097 m/z (Calc): 180.0965 Diff (ppm): 2.77 = p.cresol (TMS der)

Analysis of Biomarkers in Crude Oil

• Biomarkers: more stable against weathering

• Include: (heterocyclic) PAHs, hopanes, steranes (can be > 100 analytes)

• Analysed for oil characterization, oil spillage identification,…

• Classical method: fractionation (SPE, HPLC) + GC-MS (EI) using multiple SIM methods

Agilent 7200 GC/Q-TOF conditions

• 1 µL splitless injection of 1 mg/mL oil sample

• 30 m x 0.25 mm ID x 0.25 µm DB-5MS

• 1.5 mL/min helium, constant flow

• 50°C (1 min) – 10°C/min – 320°C (8 min)

• MS:

– 40 – 500 Da acquisition, 5 Hz

– Source 280°C, quad: 150°C

PAH mix with dibenzothiophene (0.38 ng OC)

NIST 2260 mixture DBT = C12H8S 184.0341

Number 1 hit = DBT

Analysis of DBT – Mass Accuracy

Mass error: - 1.09 ppm 184.0339

April 8, 2012 27

Analysis of Biomarkers in Crude Oil

April 8, 2012 28

Analysis of Biomarkers in Crude Oil

Mass extraction window: +/- 0.5 amu Equivalent to MSD

EIC 184

Interferences

dibenzothiophene

April 8, 2012 29

Analysis of Biomarkers in Crude Oil

Mass extraction window: +/- 5 ppm

EIC 184.0341 dibenzothiophene

April 8, 2012 30

Analysis of DBT – Mass Accuracy (in matrix)

Mass error in matrix: - 1.63 ppm

184.0338

DBT, mono- and dimethyl DBTs

184.0341

198.0498

212.0645

Hopanes

(mainlib) 28-Nor-17 a( H)-hopane

100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400 410

0

50

100

109 123

135

149

163

177

191

205

217

245 260

355 383 398

Analysis of Hopanes in Crude Oil

191 +/- 0.5

191.1794 +/- 10 ppm

Interferences hopanes

hopanes

Confidentiality Label

April 8, 2012 34

Analysis of Steranes in Crude Oil

EIC 217.1951

EIC 400.4064

Confidentiality Label

April 8, 2012 35

Steranes in Crude Oil by GC-Q-TOF - MS/MS mode

MS: EIC 400.4064

MS/MS: 400 > 217.1951

Conclusions

• Mass accuracy and resolution: excellent

• Sensitivity: higher than expected

• Stability: excellent

• Outlook:

– Excellent tool for compound ID (“beyond EI MS library search”)

– “open methods” – “retrospective analysis”