TOF Analysis of Bay Water surrounding the

Battleship Texas

Presented at: SETAC 2013 South Regional Conference

Work By: Sue D’Antonio, Joan Stevens, Andre Szczesniewski , Lynne Marshall, John Kimmel and Ken Helmreich

Agilent 6230 TOF LC/MS

TOF and QTOF techniques (non-target compounds)• Identifies and differentiates compounds that were

previously “hidden” under background• Accurate mass TOF instruments:

– Can determine empirical formula based on the mass of the compound in question (accurate mass)

Novel data mining techniquesAccurate Mass Databases, Mass Profile Processional Software • Discover more compounds/contaminants, faster

and with greater precision

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API-TOF Ion Optical Design

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API Ion Source

Beam Shaper

TOF Mass Analyzer

Two StageIon Mirror

Optically CoupledIon Detector

Effective Flight PathLength of 2.0m

Formulae for TOF Performance

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Resolution = (m/z)/w1/2

1000/0.1 = 10,000

922/0.087 = 10,597

Mass accuracy (mass error)

(1000.001-1000.000)/1000)x106=1 ppm

Therefore: 1 ppm @ 1000 m/z is 0.001Da (1 mDa)

921.5 922.0 922.5 923.0 923.5 924.0 924.5m/z, amu

923.0121

W1/2 = 0.087

+TOF MS: 5.474 to 5.547 min from steroid2TOFap42.wiff Agilent, subtracted (4.696 to ... Max. 1.2e5 counts.

120 140 160 180 200 220 240 260 280 300 320 340 360 380 400m/z, amu

0.00

1.00e4

2.00e4

3.00e4

4.00e4

5.00e4

6.00e4

7.00e4

8.00e4

9.00e4

1.00e5

1.10e5

1.16e5

Inte

ns

ity, c

ou

nts

289.2159

271.2051

269.1893

287.1997

Epitestosterone

Resolution 6890

meas = 289.2159

actual = 289.2162

-1.06 ppm

922.0092

610actual

actual - measured accuracy Mass ´÷øö

çèæ=

How Much Accuracy is Needed?(Newer concept for LC/MS: ppm accuracy)

Reserpine (C33H40N2O9) has a protonated ion at 609.28066

Single quad reports mass to +/- 0.1 = 165 ppm

Number of possible formulas using only C, H, O & N:• 165 ppm209• 10 ppm 13• 5 ppm 7• 3 ppm 4• 2 ppm 2

Accurate mass reduces number of candidates and risk of investing effort with the wrong molecule

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6

SPE Sample Prep

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Analysis of Water Samples

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Choose Your Elements: Find Compounds by Molecular Feature and Generate Formulae

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Molecular Feature Generator

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Compound Chromatogram

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Deductive Compound Choices

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Conclusions

Utilizing Agilent’s Time of Flight accurate mass, high resolution, superior isotopic fidelity and the unique features of Mass Hunter software, such as Molecular Feature Extractor and Molecular Formula Generator, we were able to identify over 100 compounds found in the samples.

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