Abigail Sundberg1, Paolo Benigni1, Lilian Valadares Tose1, Dennys Leyva1,

Cesar Ramirez1, Rudolph Jaffe1, and Francisco Fernandez-Lima1,2

1Department of Chemistry and Biochemistry, Florida International University, Miami, USA2 Biomolecular Science Institute, Florida International University, Miami, USA

State of the Art MS

Overview

Experimental Methods

Introduction

Acknowledgments

Experimental Results

ConclusionsTIMS provides fast gas phase separation of isomers, as is shown. Coupling LC with TIMS

increases the separation potential of the TIMS by adding another dimension of separation for

structures with similar collision cross sections. Utilizing LC-TIMS-ToF MS gives

complementary orthogonal separation of the isomers. We compared the number of features

identified by ToF MS and FT-ICR and found that the ToF provided primary intense features,

but the FT-ICR provided more features overall and allowed for the determination of chemical

formulas. Next step is to further evaluate the analytical capabilities using TIMS FT-ICR MS.

Experimental Results

Ionization

Source

to MS

Vplate Ventrance Vramp Vout

P2

FE Fdrift

P1 > P2

LC and Mobility Separation – ToF MS

This material is based upon work supported by the National Science Foundation

under Grant No. HRD-1547798. This NSF Grant was awarded to Florida International

University as part of the Centers of Research Excellence in Science and Technology

(CREST) Program. Any opinions, findings, and conclusions or recommendations expressed

in this material are those of the author(s) and do not necessarily reflect the views of the

National Science Foundation.

The authors acknowledge the financial support from the National Institute of Health

(R00GM106414), the National Science Foundation Division of Chemistry, under CAREER

award CHE-1654274, with co-funding from the Division of Molecular and Cellular Biosciences

to FFL.

200 400 600 800

10

20

30

40

Pea

ks

per

nom

inal

mas

s

m/z

FT-ICR MS

LC-ToF MS

0.0 0.2 0.4 0.6 0.8 1.0 1.20.20.40.60.81.01.21.41.61.82.02.2

H /

C

O / C0.0 0.2 0.4 0.6 0.8 1.0 1.2

0.20.40.60.81.01.21.41.61.82.02.2

H /

C

O / C

Fernandez-Lima F, Blase RC, Russell DH. 2011. International Journal of Mass Spectrometry 298:111-8

Fernandez-Lima F, Kaplan DA, Park MA. 2011. Review of Scientific Instruments 82:126106

Fernandez-Lima F, Kaplan DA, Suetering J, Park MA. 2011. International Journal for Ion Mobility Spectrometry 14:93-8

SWRFA LC-ToF MS vs FT-ICR MS

LC-TIMS MS SWRFA

Figure 1.Schematics of the FT-ICR MS instrument used to analyze the SWRFA

standard

Figure 2.Schematics of the TIMS used to analyze the all samples

Figure 3.Schematics of the TIMS-ToF instrument used to analyze all samples

145 150 155 160 165 170

Rel

ativ

e In

tensi

ty

CCS (Å2)

145 150 155 160 165 170

Rel

ativ

e In

tensi

ty

CCS (Å2)

145 150 155 160 165 170

Rel

ativ

e In

tensi

ty

CCS (Å2)

156.5 157.0 157.5 158.0 158.5 159.0 159.5 160.0 160.5

Rel

ativ

e In

ten

sity

CCS (Å2)

156 157 158 159 160

Rel

ativ

e In

tensi

ty

CCS (Å2)

Fernandez-Lima F, Blase RC, Russell DH. 2011. International Journal of Mass Spectrometry 298:111-8

Fernandez-Lima F, Kaplan DA, Park MA. 2011. Review of Scientific Instruments 82:126106

Fernandez-Lima F, Kaplan DA, Suetering J, Park MA. 2011. International Journal for Ion Mobility Spectrometry 14:93-8

Figure 6. a-c) 2D LC-IMS contour maps of three structural isomers for m/z 201.06 using LC-TIMS-

ToF MS. Notice the orthogonality between the LC and IMS separations.

Figure 7. a) Typical ESI negative mode mass spectra

of SWRFA standard using LC-ToF MS (green) and

FT-ICR (blue), and b) Close up of m/z 407.

c) The number of peaks per nominal mass for the LC-

ToF MS (green) and FT-ICR MS (blue), (g, d-e) Van

Krevelen plots using LC-ToF MS (Left) and FT-ICR

MS (Right)

Notice that the LC-ToF MS spectrum is able to identify

the primary features of the SWRFA but the FT-ICR is

able to resolved a greater number of features.

Dissolved organic matter (DOM) is present in many aquatic ecosystems and is

important for many biological systems. With the recent development of Trapped

Ion Mobility Spectrometry (TIMS) and its coupling to time of flight (ToF) and

Fourier Transform Ion Cyclotron Resonance (FT-ICR) mass spectrometries (MS),

in this project we evaluate new analytical workflows for the analysis of DOM. In

addition, the coupling of pre-separaton techniques based on liquid

chromatography (LC) were evaluated to reduce sample complexity during

ionization and increase the orthogonality of the separation. The advantages of

the LC-TIMS-ToF MS platform were illustrated for the separation of structural

isomers and a complex mixture of DOM. Results show that the high mobility

resolving power of the TIMS analyzer permits the separation of structural isomers

and allowed determining accurate collision cross sections (CCS). The analysis of

the Suwanee River Fulvic Acid (SWRFA) by LC-TIMS-ToF MS provided new

insights into the structural complexity of DOM mixtures and opened new avenues

for further exploration of LC separations.

Dissolved organic matter (DOM) is present in many both salt water and fresh

water ecosystems and is used by microorganisms to support aquatic

ecosytems.1 DOM also acts as a filter for sunlight that passes through water,

protecting aquatic species.2 There are varying types of compounds, including

aromatic compounds, that make up the composition of DOM samples.3 There

can be some similarities between different DOM samples collected from different

locations or from the same location during different hydroperiods. However, there

are distinct differences, indicating that there can be different biogeochemical

processes taking place or different influences from the environment.4 Previous

work conducted by Norbert Hertkorn, Mourad Harir, Kaelin Cawley, and Rudolf

Jaffe containing proton nuclear magnetic resonance spectrometry (1H NMR) and

fourier transform ion cyclotron resonance (FT-ICR) data for different DOM

standards provided a reference for selecting the isomeric standards used in this

project.4 In the present work, we further expand the analysis of DOM by coupling

LC-TIMS, adding orthogonal separations prior to mass analysis.

Experimental Methods

Experimental Results

200 202 204 206 208

Rel

ativ

e In

tensi

ty

m/z

[M-H]-

159 A2

R=60

157 A2

R=61

159 A2

R=67

3-Methoxy-2-naphthoic acid

2-Methoxy-1-naphthoic acid

4-Methoxy-1-naphthoic acid 2M1NA

4M1NA

2M1NA

C12H10O3

m/z 201.056

Mobility Separation – ToF MS

Figure 5. a-c) Typical low resolution mobility spectra for representative DOM

isomers. d) Low resolution separation of a binary mixture of structural isomers, and

e.) high resolution separation of a ternary mixture of isomers.

Figure 4.

Typical mass

spectrum for all

of the structural

isomers.

3M2NA

3M2NA

a)

b)

c)

d)

e)

a) b) c)

b)a)

c)

d) e)

Figure 8. a) 2D IMS-MS contour plot of the SWRFA standard

within m/z 200 to 800. In the insets (b-d) 2D LC-IMS contour

plots are shown for individual chemical formulas within a

nominal mass.

3 8 12 16 21 25 29 33

148

128

107

20.5

575.5

3 8 12 16 21 25 29 33

148

128

107

20.5

575.5

m/z 200.8987

m/z 201.0991

[C12H10O3- H]-

[C12H10O3- H]-

b)

c)

d)

a)

1Lawrence R. Pomeroy. Bioscience, Vol. 24, No. 9. Sept., 1974. 499-5042Kenneth S. Johnson, Kenneth H. Coale, and Hans W. Jannasch. Analytical Chemistry, Vol. 64, No.22, Nov. 15,

1992. 1065-1075A3Chen, M., Maie, N., Parish, K., and Jaffé, R., Bio-geochemistry, Vol. 115, 2013. 167–1834Norbert Hertkorn, Mourad Harir, Kaelin Cawley, Rudolf Jaffe. Biogeosciences Discuss., Vol. 12, 2015. 13711-13765

m/z 201.0259[C8H10O6-H]-

Abigail Sundberg, asund006@fiu.edu

RT (min)

CC

S (Å

2)C

CS

(Å2)

CC

S (Å

2)