Analysis of Organic Anionic Surfactants in Fine and Coarse Fractions
Supporting Information
Analysis of Organic Anionic Surfactants in Fine and Coarse Fractions of Freshly Emitted Sea Spray Aerosol Richard E. Cochran1, Olga Laskina1, Thilina Jayarathne1, Alexander Laskin2, Julia Laskin3, Peng Lin2, Camille Sultana4, Christopher Lee4, Kathryn A. Moore4, Christopher D. Cappa5, Timothy H. Bertram6, Kimberly A. Prather4,7, Vicki H. Grassian*,4,7, Elizabeth A. Stone*,1 1
Department of Chemistry, University of Iowa, Iowa City, Iowa, USA William R. Wiley Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington, USA 3 Physical Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA 4 Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA 5 Department of Civil and Environmental Engineering, University of California, Davis, Davis, California, USA 6 Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA 7 Scripps Institution of Oceanography, University of California, San Diego, La Jolla, California, USA 2
* Corresponding authors: Vicki H. Grassian, [email protected]. Tel: 858-534-2499, Fax: 858-534-6255 Elizabeth A. Stone, [email protected]. Tel: 319-384-1863, Fax: 319-335-1270
Summary of Contents This 14-page document contains 5 figures and 3 tables: Daily mass and OC concentrations for fine and coarse SSA particles during the mesocosm experiment (Figure S1), total and extracted ion chromatograms for standards and samples (Figure S2), signal distributions for FA and FA derivatives (Figure S3), reaction pathways leading to the formation of FA sat and FAunsat (Figure S4), average molecular weights for select homologous series (Figure S5), descriptions of the mesocosm experiment and biological data for the five sampling periods (Table S1), HRMS data used for the identification of organic compounds (Table S2), relative intensities of FA and FA derivatives (Table S3).
S1
Figure S1. Daily measured total mass (a) and organic carbon mass (b) concentration in fine (Dp ≤ 2.5 µm) and coarse (Dp = 2.5–10 µm) SSA plotted over the observed levels of chlorophyll-a during the mesocosm experiment. All samples were collected daily during the experiment. Daily samples were combined (between 5-6 days) into a single sample prior to analysis, resulting in five separate “periods” of the mesocosm experiment (labels shown). Chlorophyll-a data were replotted from Reference 14.
S2
Figure S2. The tentative assignment of compounds to observed masses in the ESI-LTQ-Orbitrap spectra were further supported by comparing retention times in the UPLC-ESI-QToF to authentic standards as well as the relative retention times in each homologous series. An extracted ion chromatogram (EIC) from the UPLC-ESI-QToFMS analysis a mixture of authentic standards, each at 5 ppm (a), shows the retention times of representative saturated fatty acids, saturated hydroxy-fatty acids and saturated dicarboxylic acids. The total ion chromatogram (TIC) obtained from the UPLC-ESI-QToFMS analysis of the extract of fine SSA collected during Period 5 of the mesocosm is shown in (b). Each homologous series of biogenic anionic surfactants observed in the fine SSA are represented by EICs: (c) saturated fatty acids, (d) monounsaturated fatty acids, (e) diunsaturated fatty acids, (f) saturated oxo-fatty acids and (g) saturated hydroxy-fatty acids. EICs are also shown for anthropogenic linear alkylsulfates (h) and linear alkylbenzenesulfonates (i). Each EIC curve is traced from the measured responses of ions that have a mass within ±0.01 Da of the theoretical mass (the inset of each EIC shows the masses as well as their assigned empirical formulas).
S3
Figure S2 continued.
S4
Figure S2. continued
S5
1 2 3 4 5 6
Figure S3. Distribution of FAs and FA derivatives observed in compartments during the mesocosm waveflume experiment. The full scale distribution is shown in (a) and a zoomed-in scale in (b). The identity of the FA species represented by each bar is shown by the short-hand notation (e.g., hexadecanoic acid, “C16:0”). Responses of each individual compound were obtained from composite HRMS spectra from UPLC-ESI-QToF analysis, with ESI in negative mode.
S6
Figure S4. Reaction pathways leading to the formation of (a) FAsat and FAunsat from cellular components (phospholipids and triacylglycerides) of phytoplankton and bacteria as well as (b) FAsat and FAOH from lipopolysaccharides present in heterotrophic bacteria (gram negative). The reaction pathway for the oxidation of FAunsat, producing 9-oxononanoic acid, is also shown in (a). The pathways shown are proposed to be the major sources of both FAsat and FAunsat as well as FAoxo that were observed in nascent SSA.
S7
Figure S5. Average molecular weights of a) FAunsat, b) FAoxo, c) FAOH and d) saturated dicarboxylic acids within fine (Dp ≤ 2.5 µm), coarse (Dp = 2.5–10 µm) SSA as well as the SSML and SSW compartments. All samples were collected daily during the mesocosm experiment. Daily samples were combined (between 5-6 days) into a single sample prior to analysis, resulting in five “periods” of the mesocosm experiment. Samples were analyzed with UPLC-ESI-QToF MS in negative mode.
S8
Table S1. Details of the sampling of nascent SSA as well as online measurements of biological tracers in sub-surface seawater during the mesocosm experiment. Mesocosm Time Period
a
Dates in Period
Mesocosm Experiment Activity
1
7/5–10
2
7/11–15
3
7/16–19
4
7/20–23
5
7/24–29
System equilibration First half of phytoplankton bloom #1 Second half of phytoplankton bloom #1 Between Phytoplankton blooms Phytoplankton bloom #2
Fine SSA Mass Percent Concentration OC (µg/m3) (%)
Coarse SSA Mass Percent Concentration OC (µg/m3) (%)
Levels of Markers for Biological Activity Chlorophyll-a Heterotrophic (Phytoplankton) Bacteria (µg/L)a (counts, 103)a
28
±
10
28
±
5
79
±
28
3
±
1
1.1
±
0.4
6.1
±
3.2
34
±
11
12
±
3
206
±
74
1.1
±
0.3
8.3
±
6.8
7.5
±
4.3
52
±
15
7
±
1
243
±
75
1.0
±
0.2
17.7
±
6.1
11
±
12
55
±
15
6
±
1
179
±
57
1.4
±
0.3
7.1
±
5.9
26
±
3.6
52
±
17
6
±
2
192
±
73
1.2
±
0.4
24.1
±
3.6
31
±
25
Values are shown as the mean ± one standard deviation.
S9
Table S2. Data used for the identification of organic compounds using ESI-LIT-Orbitrap MS. Tentatively Identified Compounda,b
Carbon Number
Empirical Formula
DBE
Monoisotopic Mass (Da) Theoretical Measurede
Error (ppm)
8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
C8H16O2 C9H18O2 C10H20O2 C11H22O2 C12H24O2 C13H26O2 C14H28O2 C15H30O2 C16H32O2 C17H34O2 C18H36O2 C19H38O2 C20H40O2 C21H42O2 C22H44O2 C23H46O2 C24H48O2
1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
143.10775 157.12340 171.13905 185.15470 199.17035 213.18600 227.20165 241.21730 255.23295 269.24860 283.26425 297.27990 311.29555 325.31120 339.32685 353.34250 367.35815
143.10766 157.12329 171.13892 185.15457 199.17013 213.18581 227.20139 241.21703 255.23265 269.24827 283.26390 297.27954 311.29512 325.31080 339.32640 353.34201 367.35765
-0.65 -0.74 -0.81 -0.73 -1.12 -0.92 -1.16 -1.14 -1.21 -1.23 -1.25 -1.24 -1.41 -1.24 -1.34 -1.40 -1.37
12 13 14 15 16 17 18 12 13 14 15 16 17 18 14 16 18 11 12 16 18 20 21 18 20 22 22
C12H22O2 C13H24O2 C14H26O2 C15H28O2 C16H30O2 C17H32O2 C18H34O2 C12H20O2 C13H22O2 C14H24O2 C15H26O2 C16H28O2 C17H30O2 C18H36O2 C14H22O2 C16H26O2 C18H30O2 C11H14O2 C12H16O2 C16H24O2 C18H28O2 C20H32O2 C21H34O2 C18H26O2 C20H30O2 C22H34O2 C22H32O2
2 2 2 1 2 2 2 3 3 3 3 3 3 3 4 4 4 5 5 5 5 5 5 6 6 6 7
197.15470 211.17035 225.18600 239.20165 253.21730 267.23295 281.24860 195.13905 209.15470 223.17035 237.18600 251.20165 265.21730 279.23295 221.15470 249.18600 277.21730 177.09210 191.10775 247.17035 275.20165 303.23295 317.24860 273.18600 301.21730 329.24860 327.23295
197.15455 211.17025 225.18580 239.20142 253.21700 267.23264 281.24824 195.13888 209.15455 223.17015 237.18579 251.20134 265.21700 279.23258 221.15455 249.18570 277.21700 177.09203 191.10762 247.17010 275.20135 303.23255 317.24820 273.18575 301.21700 329.24816 327.23245
-0.78 -0.49 -0.90 -0.99 -1.20 -1.17 -1.29 -0.91 -0.73 -0.91 -0.90 -1.27 -1.14 -1.34 -0.69 -1.22 -1.09 -0.42 -0.70 -1.03 -1.10 -1.33 -1.27 -0.93 -1.02 -1.35 -1.54
Saturated Fatty Acids octanoic acid nonanoic acid decanoic acid undecanoic acid dodecanoic acid tridecanoic acid tetradecanoic acid pentadecanoic acid hexadecanoic acid heptadecanoic acid octadecanoic acid nonadecanoic acid eicosanoic acid uneicosanoic acid dodeicosanoic acid trieicosanoic acid tetracosanoic acid Unsaturated Fatty Acids dodecenoic acid tridecenoic acid tetradecenoic acid pentadecenoic acid hexadecenoic acid heptadecenoic acid octadecenoic acid dodecadienoic acid tridecadienoic acid tetradecadienoic acid pentadecadienoic acid hexadecadienoic acid heptadecadienoic acid octaadecadienoic acid tetradecatrienoic acid hexadecatrienoic acid octadecatrienoic acid undecatetraenoic acid dodecatetraenoic acid hexadecatetraenoic acid octadecatetraenoic acid eicosatetraenoic acid uneicosatetraenoic acid octadecapentaenoic acid eicosapentaenoic acid doeicosapentaenoic acid doeicosahexaenoic acid Saturated Oxo-Fatty Acids
S10
oxopentanoic acid oxohexanoic acid oxoheptanoic acid oxooctanoic acid oxononanoic acid oxodecanoic acid oxoundecanoic acid oxododecanoic acid oxotridecanoic acid oxotetradecanoic acid oxopentadecanoic acid oxohexadecanoic acid oxoheptadecanoic acid oxooctadecanoic acid
5 6 7 8 9 10 11 12 13 14 15 16 17 18
C5H8O3 C6H10O3 C7H12O3 C8H14O3 C9H16O3 C10H18O3 C11H20O3 C12H22O3 C13H24O3 C14H26O3 C15H28O3 C16H30O3 C17H32O3 C18H34O3
2 2 2 2 2 2 2 2 2 2 2 2 2 2
115.04007 129.05572 143.07137 157.08702 171.10267 185.11832 199.13397 213.14962 227.16527 241.18092 255.19657 269.21222 283.22787 297.24352
115.03996 129.05562 143.07132 157.08694 171.10256 185.11817 199.13380 213.14939 227.16505 241.18070 255.19640 269.21192 283.22752 297.24315
-0.94 -0.79 -0.36 -0.51 -0.66 -0.79 -0.85 -1.07 -0.96 -0.91 -0.66 -1.11 -1.25 -1.24
5 6 7 8 9 10 11 12 13 14 15 16 17 18
C5H10O3 C6H12O3 C7H14O3 C8H16O3 C9H18O3 C10H20O3 C11H22O3 C12H24O3 C13H26O3 C14H28O3 C15H30O3 C16H32O3 C17H34O3 C18H36O3
1 1 1 1 1 1 1 1 1 1 1 1 1 1
117.05572 131.07137 145.08702 159.10267 173.11832 187.13397 201.14962 215.16527 229.18092 243.19657 257.21222 271.22787 285.24352 299.25917
117.05567 131.07130 145.08696 159.10262 173.11818 187.13383 201.14946 215.16506 229.18072 243.19628 257.21194 271.22763 285.24322 299.25891
-0.41 -0.52 -0.44 -0.30 -0.80 -0.74 -0.81 -0.97 -0.87 -1.21 -1.08 -0.88 -1.05 -0.86
3 4 5 6 7 8 9 10 11 12 13 14 15 16 18 19 20 22
C3H4O4 C4H6O4 C5H8O4 C6H10O4 C7H12O4 C8H14O4 C9H16O4 C10H18O4 C11H20O4 C12H22O4 C13H24O4 C14H26O4 C15H28O4 C16H30O4 C18H34O4 C19H36O4 C20H38O4 C22H42O4
2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2
103.00368 117.01933 131.03498 145.05063 159.06628 173.08193 187.09758 201.11323 215.12888 229.14453 243.16018 257.17583 271.19148 285.20713 313.23843 327.25408 341.26973 369.30103
103.00356 117.01922 131.03489 145.05054 159.06616 173.08178 187.09740 201.11306 215.12867 229.14431 243.15997 257.17562 271.19149 285.20684 313.23808 327.25368 341.26922 369.30059
-1.17 -0.98 -0.71 -0.66 -0.75 -0.90 -0.99 -0.88 -0.99 -0.97 -0.88 -0.83 0.02 -1.03 -1.12 -1.23 -1.52 -1.20
3
C3H8O4S
0
139.00705
139.00696
-0.68
Saturated Hydroxy-Fatty Acids hydroxy-pentanoic acid hydroxy-hexanoic acid hydroxy-heptanoic acid hydroxy-octanoic acid hydroxy-nonanoic acid hydroxy-decanoic acid hydroxy-undecanoic acid hydroxy-dodecanoic acid hydroxy-tridecanoic acid hydroxy-tetradecanoic acid hydroxy-pentadecanoic acid hydroxy-hexadecanoic acid hydroxy-heptadecanoic acid hydroxy-octadecanoic acid Saturated Dicarboxylic Acidsc propanedioic acid butanedioic acid pentanedioic acid hexanedioic acid heptanedioic acid octanedioic acid nonanedioic acid decanedioic acid undecanedioic acid dodecanedioic acid tridecanedioic acid tetradecanedioic acid pentadecanedioic acid hexadecanedioic acid octadecanedioic acid nonadecanedioic acid eicosanedioic acid doeicosanedioic acid Sulfates monopropylsulfate
S11
monotetrasulfate monopentasulfate monohexylsulfate monoheptasulfate monooctylsulfate monononasulfate monodecylsulfate monoundecylsulfate monododecylsulfate monotridecylsulfate monotetradecylsulfate monopentadecylsulfate monohexadecylsulfate monoheptadecylsulfate
4 5 6 7 8 9 10 11 12 13 14 15 16 17
C4H10O4S C5H12O4S C6H14O4S C7H16O4S C8H18O4S C9H20O4S C10H22O4S C11H24O4S C12H26O4S C13H28O4S C14H30O4S C15H32O4S C16H34O4S C17H36O4S
0 0 0 0 0 0 0 0 0 0 0 0 0 0
153.02270 167.03835 181.05400 195.06965 209.08530 223.10095 237.11660 251.13225 265.14790 279.16355 293.17920 307.19485 321.21050 335.22615
153.02258 167.03821 181.05384 195.06945 209.08531 223.10075 237.11628 251.13202 265.14756 279.16319 293.17879 307.19445 321.21007 335.22567
-0.81 -0.87 -0.91 -1.05 0.03 -0.92 -1.39 -0.95 -1.30 -1.31 -1.41 -1.32 -1.35 -1.45
16 17 18 19 20
C16H26O3S C17H28O3S C18H30O3S C19H32O3S C20H34O3S
4 4 4 4 4
297.15299 311.16864 325.18429 339.19994 353.21559
297.15257 311.16816 325.18374 339.19936 353.21501
-1.41 -1.54 -1.69 -1.71 -1.64
Linear Alkylbenzene Sulfonatesd decylbenzenesulfonate undecylbenzenesulfonate dodecylbenzenesulfonate tridecylbenzenesulfonate tetradecylbenzenesulfonate a
The empirical formulas for the proposed linear chain acids may also correspond to branched chain acids as well.
b
All monoisotopic masses shown correspond to the deprotonated ion. Empirical formulas for the tentatively identified dicarboxylic acids may also correspond to saturated oxo-hydroxycarboxylic acids. d Stereo-connectivity are not shown as they commonly exist as mixtures of species with the benzenesulfonate at various positions of the alkyl chain. e The measured masses shown were obtained from the analysis of fine SSA collected during time period 3. c
S12
Table S3. Relative intensities of fatty acids and fatty acid derivatives tentatively identified during the mesocosm experiment. Values were calculated based on the signal intensities of the [M-H]- ion for each compound in composite spectra from UPLC-ESI-QToF MS analysis. Acid Class
Acid Name (Tentative)
Saturated Fatty Acids octanoic acid nonanoic acid decanoic acid undecanoic acid dodecanoic acid tridecanoic acid tetradecanoic acid pentadecanoic acid hexadecanoic acid heptadecanoic acid octadecanoic acid nonadecanoic acid eicosanoic acid uneicosanoic acid dodeicosanoic acid trieicosanoic acid tetracosanoic acid
Carbon #
Degree of Unsat.
SSW
8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
0.2 1 0.7 0.2 1 0.4 4 2 34 3 39 0.1 0.1 0.04 0.1 NRa 0.05
1 2 3 0.9 3 1.3 5 4 25 3 28 0.1 0.7 2 2 0.1 0.2
0.2 0.3 0.3 0.2 2 1.1 6 3 32 3 38 0.3 0.2 0.2 0.1 0.03 0.1
0.2 0.6 0.6 0.2 2 1.9 8 4 32 3 30 0.2 0.1 0.04 0.1 0.02 NR
86
81
86
84
0.01 0.02 0.05 0.05 0.4 0.1 1 0.09 0.03 0.02 0.03 0.02 0.01 0.3 0.1 0.01 0.03 0.05 0.06 0.04 0.02 0.05 0.01 0.03 0.1 0.04
0.03 0.02 0.1 0.1 1 0.3 0.8 0.2 0.05 0.03 0.01 0.3 0.03 2 0.5 0.02 0.01 0.01 NR 0.02 0.01 0.05 NR 0.02 0.3 NR
0.03 0.04 0.07 0.09 0.3 0.2 0.9 0.02 0.02 0.03 0.03 0.06 0.1 0.5 0.1 0.01 0.1 0.04 0.04 0.02 0.03 0.1 0.1 0.07 0.2 0.1
0.04 0.05 0.1 0.2 0.4 0.2 0.6 0.1 0.05 0.05 0.1 0.1 0.07 0.2 0.1 0.02 0.06 0.06 0.07 0.04 0.01 0.04 0.03 0.02 0.09 0.02
Sum of Saturated Fatty Acids Unsaturated Fatty Acids dodecenoic acid tridecenoic acid tetradecenoic acid pentadecenoic acid hexadecenoic acid heptadecenoic acid octadecenoic acid dodecadienoic acid tridecadienoic acid tetradecadienoic acid pentadecadienoic acid hexadecadienoic acid heptadecadienoic acid octaadecadienoic acid tetradecatrienoic acid hexadecatrienoic acid octadecatrienoic acid undecatetraenoic acid dodecatetraenoic acid hexadecatetraenoic acid octadecatetraenoic acid eicosatetraenoic acid uneicosatetraenoic acid octadecapentaenoic acid eicosapentaenoic acid doeicosapentaenoic acid
12 13 14 15 16 17 18 12 13 14 15 16 17 18 14 16 18 11 12 16 18 20 21 18 20 22
1 1 1 1 1 1 1 2 2 2 2 2 2 2 3 3 3 4 4 4 4 4 4 5 5 5
S13
Percent of Total FA Signal SSML Coarse SSA
Fine SSA
doeicosahexaenoic acid
22
6
Sum of Unsaturated Fatty Acids Oxygenated Saturated Fatty Acids oxopentanoic acid oxohexanoic acid oxoheptanoic acid oxooctanoic acid oxononanoic acid oxodecanoic acid oxoundecanoic acid oxododecanoic acid oxotridecanoic acid oxotetradecanoic acid oxopentadecanoic acid oxohexadecanoic acid oxoheptadecanoic acid oxooctadecanoic acid
5 6 7 8 9 10 11 12 13 14 15 16 17 18
0 0 0 0 0 0 0 0 0 0 0 0 0 0
Sum of Saturated Oxo-Fatty Acids Hydroxylated Saturated Fatty Acids hydroxy-pentanoic acid hydroxy-hexanoic acid hydroxy-heptanoic acid hydroxy-octanoic acid hydroxy-nonanoic acid hydroxy-decanoic acid hydroxy-undecanoic acid hydroxy-dodecanoic acid hydroxy-tridecanoic acid hydroxy-tetradecanoic acid hydroxy-pentadecanoic acid hydroxy-hexadecanoic acid hydroxy-heptadecanoic acid hydroxy-octadecanoic acid
5 6 7 8 9 10 11 12 13 14 15 16 17 18
0 0 0 0 0 0 0 0 0 0 0 0 0 0
Sum of Saturated Oxo-Fatty Acids a
0.02
0.02
0.07
0.04
3
6
4
3
1 0.1 0.0 0.03 0.3 0.05 0.05 0.06 0.03 0.04 0.1 0.23 0.01 0.4
0.09 0.1 0.1 0.02 0.1 0.1 0.08 0.08 0.04 0.08 0.1 0.25 0.01 0.6
0.2 0.1 0.06 0.1 0.3 0.1 0.09 0.09 0.06 0.07 0.06 0.2 0.03 0.4
0.8 0.1 0.09 0.2 0.5 0.3 0.2 0.2 0.1 0.1 0.2 0.3 0.04 0.4
3
2
2
4
0.02 0.09 0.1 0.07 0.1 0.0 0.03 0.1 0.06 0.05 NR 0.2 0.003 NR
0.1 0.07 0.08 0.08 0.08 0.07 0.05 0.0 0.07 0.09 0.01 0.3 0.03 0.2
0.03 0.02 0.02 0.06 0.04 0.03 0.03 0.04 0.01 0.02 0.01 0.1 0.03 0.1
0.04 0.04 0.04 0.09 0.1 0.08 0.05 0.08 0.004 0.1 0.01 0.2 0.02 0.2
0.8
1
0.6
1
"NR" denotes the HRMS response in the samples was below that observed in the field blanks.
S14
Analysis of Organic Anionic Surfactants in Fine and Coarse Fractions of Freshly Emitted Sea Spray Aerosol Richard E. Cochran1, Olga Laskina1, Thilina Jayarathne1, Alexander Laskin2, Julia Laskin3, Peng Lin2, Camille Sultana4, Christopher Lee4, Kathryn A. Moore4, Christopher D. Cappa5, Timothy H. Bertram6, Kimberly A. Prather4,7, Vicki H. Grassian*,4,7, Elizabeth A. Stone*,1 1
Department of Chemistry, University of Iowa, Iowa City, Iowa, USA William R. Wiley Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington, USA 3 Physical Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA 4 Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA 5 Department of Civil and Environmental Engineering, University of California, Davis, Davis, California, USA 6 Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA 7 Scripps Institution of Oceanography, University of California, San Diego, La Jolla, California, USA 2
* Corresponding authors: Vicki H. Grassian, [email protected]. Tel: 858-534-2499, Fax: 858-534-6255 Elizabeth A. Stone, [email protected]. Tel: 319-384-1863, Fax: 319-335-1270
Summary of Contents This 14-page document contains 5 figures and 3 tables: Daily mass and OC concentrations for fine and coarse SSA particles during the mesocosm experiment (Figure S1), total and extracted ion chromatograms for standards and samples (Figure S2), signal distributions for FA and FA derivatives (Figure S3), reaction pathways leading to the formation of FA sat and FAunsat (Figure S4), average molecular weights for select homologous series (Figure S5), descriptions of the mesocosm experiment and biological data for the five sampling periods (Table S1), HRMS data used for the identification of organic compounds (Table S2), relative intensities of FA and FA derivatives (Table S3).
S1
Figure S1. Daily measured total mass (a) and organic carbon mass (b) concentration in fine (Dp ≤ 2.5 µm) and coarse (Dp = 2.5–10 µm) SSA plotted over the observed levels of chlorophyll-a during the mesocosm experiment. All samples were collected daily during the experiment. Daily samples were combined (between 5-6 days) into a single sample prior to analysis, resulting in five separate “periods” of the mesocosm experiment (labels shown). Chlorophyll-a data were replotted from Reference 14.
S2
Figure S2. The tentative assignment of compounds to observed masses in the ESI-LTQ-Orbitrap spectra were further supported by comparing retention times in the UPLC-ESI-QToF to authentic standards as well as the relative retention times in each homologous series. An extracted ion chromatogram (EIC) from the UPLC-ESI-QToFMS analysis a mixture of authentic standards, each at 5 ppm (a), shows the retention times of representative saturated fatty acids, saturated hydroxy-fatty acids and saturated dicarboxylic acids. The total ion chromatogram (TIC) obtained from the UPLC-ESI-QToFMS analysis of the extract of fine SSA collected during Period 5 of the mesocosm is shown in (b). Each homologous series of biogenic anionic surfactants observed in the fine SSA are represented by EICs: (c) saturated fatty acids, (d) monounsaturated fatty acids, (e) diunsaturated fatty acids, (f) saturated oxo-fatty acids and (g) saturated hydroxy-fatty acids. EICs are also shown for anthropogenic linear alkylsulfates (h) and linear alkylbenzenesulfonates (i). Each EIC curve is traced from the measured responses of ions that have a mass within ±0.01 Da of the theoretical mass (the inset of each EIC shows the masses as well as their assigned empirical formulas).
S3
Figure S2 continued.
S4
Figure S2. continued
S5
1 2 3 4 5 6
Figure S3. Distribution of FAs and FA derivatives observed in compartments during the mesocosm waveflume experiment. The full scale distribution is shown in (a) and a zoomed-in scale in (b). The identity of the FA species represented by each bar is shown by the short-hand notation (e.g., hexadecanoic acid, “C16:0”). Responses of each individual compound were obtained from composite HRMS spectra from UPLC-ESI-QToF analysis, with ESI in negative mode.
S6
Figure S4. Reaction pathways leading to the formation of (a) FAsat and FAunsat from cellular components (phospholipids and triacylglycerides) of phytoplankton and bacteria as well as (b) FAsat and FAOH from lipopolysaccharides present in heterotrophic bacteria (gram negative). The reaction pathway for the oxidation of FAunsat, producing 9-oxononanoic acid, is also shown in (a). The pathways shown are proposed to be the major sources of both FAsat and FAunsat as well as FAoxo that were observed in nascent SSA.
S7
Figure S5. Average molecular weights of a) FAunsat, b) FAoxo, c) FAOH and d) saturated dicarboxylic acids within fine (Dp ≤ 2.5 µm), coarse (Dp = 2.5–10 µm) SSA as well as the SSML and SSW compartments. All samples were collected daily during the mesocosm experiment. Daily samples were combined (between 5-6 days) into a single sample prior to analysis, resulting in five “periods” of the mesocosm experiment. Samples were analyzed with UPLC-ESI-QToF MS in negative mode.
S8
Table S1. Details of the sampling of nascent SSA as well as online measurements of biological tracers in sub-surface seawater during the mesocosm experiment. Mesocosm Time Period
a
Dates in Period
Mesocosm Experiment Activity
1
7/5–10
2
7/11–15
3
7/16–19
4
7/20–23
5
7/24–29
System equilibration First half of phytoplankton bloom #1 Second half of phytoplankton bloom #1 Between Phytoplankton blooms Phytoplankton bloom #2
Fine SSA Mass Percent Concentration OC (µg/m3) (%)
Coarse SSA Mass Percent Concentration OC (µg/m3) (%)
Levels of Markers for Biological Activity Chlorophyll-a Heterotrophic (Phytoplankton) Bacteria (µg/L)a (counts, 103)a
28
±
10
28
±
5
79
±
28
3
±
1
1.1
±
0.4
6.1
±
3.2
34
±
11
12
±
3
206
±
74
1.1
±
0.3
8.3
±
6.8
7.5
±
4.3
52
±
15
7
±
1
243
±
75
1.0
±
0.2
17.7
±
6.1
11
±
12
55
±
15
6
±
1
179
±
57
1.4
±
0.3
7.1
±
5.9
26
±
3.6
52
±
17
6
±
2
192
±
73
1.2
±
0.4
24.1
±
3.6
31
±
25
Values are shown as the mean ± one standard deviation.
S9
Table S2. Data used for the identification of organic compounds using ESI-LIT-Orbitrap MS. Tentatively Identified Compounda,b
Carbon Number
Empirical Formula
DBE
Monoisotopic Mass (Da) Theoretical Measurede
Error (ppm)
8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
C8H16O2 C9H18O2 C10H20O2 C11H22O2 C12H24O2 C13H26O2 C14H28O2 C15H30O2 C16H32O2 C17H34O2 C18H36O2 C19H38O2 C20H40O2 C21H42O2 C22H44O2 C23H46O2 C24H48O2
1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
143.10775 157.12340 171.13905 185.15470 199.17035 213.18600 227.20165 241.21730 255.23295 269.24860 283.26425 297.27990 311.29555 325.31120 339.32685 353.34250 367.35815
143.10766 157.12329 171.13892 185.15457 199.17013 213.18581 227.20139 241.21703 255.23265 269.24827 283.26390 297.27954 311.29512 325.31080 339.32640 353.34201 367.35765
-0.65 -0.74 -0.81 -0.73 -1.12 -0.92 -1.16 -1.14 -1.21 -1.23 -1.25 -1.24 -1.41 -1.24 -1.34 -1.40 -1.37
12 13 14 15 16 17 18 12 13 14 15 16 17 18 14 16 18 11 12 16 18 20 21 18 20 22 22
C12H22O2 C13H24O2 C14H26O2 C15H28O2 C16H30O2 C17H32O2 C18H34O2 C12H20O2 C13H22O2 C14H24O2 C15H26O2 C16H28O2 C17H30O2 C18H36O2 C14H22O2 C16H26O2 C18H30O2 C11H14O2 C12H16O2 C16H24O2 C18H28O2 C20H32O2 C21H34O2 C18H26O2 C20H30O2 C22H34O2 C22H32O2
2 2 2 1 2 2 2 3 3 3 3 3 3 3 4 4 4 5 5 5 5 5 5 6 6 6 7
197.15470 211.17035 225.18600 239.20165 253.21730 267.23295 281.24860 195.13905 209.15470 223.17035 237.18600 251.20165 265.21730 279.23295 221.15470 249.18600 277.21730 177.09210 191.10775 247.17035 275.20165 303.23295 317.24860 273.18600 301.21730 329.24860 327.23295
197.15455 211.17025 225.18580 239.20142 253.21700 267.23264 281.24824 195.13888 209.15455 223.17015 237.18579 251.20134 265.21700 279.23258 221.15455 249.18570 277.21700 177.09203 191.10762 247.17010 275.20135 303.23255 317.24820 273.18575 301.21700 329.24816 327.23245
-0.78 -0.49 -0.90 -0.99 -1.20 -1.17 -1.29 -0.91 -0.73 -0.91 -0.90 -1.27 -1.14 -1.34 -0.69 -1.22 -1.09 -0.42 -0.70 -1.03 -1.10 -1.33 -1.27 -0.93 -1.02 -1.35 -1.54
Saturated Fatty Acids octanoic acid nonanoic acid decanoic acid undecanoic acid dodecanoic acid tridecanoic acid tetradecanoic acid pentadecanoic acid hexadecanoic acid heptadecanoic acid octadecanoic acid nonadecanoic acid eicosanoic acid uneicosanoic acid dodeicosanoic acid trieicosanoic acid tetracosanoic acid Unsaturated Fatty Acids dodecenoic acid tridecenoic acid tetradecenoic acid pentadecenoic acid hexadecenoic acid heptadecenoic acid octadecenoic acid dodecadienoic acid tridecadienoic acid tetradecadienoic acid pentadecadienoic acid hexadecadienoic acid heptadecadienoic acid octaadecadienoic acid tetradecatrienoic acid hexadecatrienoic acid octadecatrienoic acid undecatetraenoic acid dodecatetraenoic acid hexadecatetraenoic acid octadecatetraenoic acid eicosatetraenoic acid uneicosatetraenoic acid octadecapentaenoic acid eicosapentaenoic acid doeicosapentaenoic acid doeicosahexaenoic acid Saturated Oxo-Fatty Acids
S10
oxopentanoic acid oxohexanoic acid oxoheptanoic acid oxooctanoic acid oxononanoic acid oxodecanoic acid oxoundecanoic acid oxododecanoic acid oxotridecanoic acid oxotetradecanoic acid oxopentadecanoic acid oxohexadecanoic acid oxoheptadecanoic acid oxooctadecanoic acid
5 6 7 8 9 10 11 12 13 14 15 16 17 18
C5H8O3 C6H10O3 C7H12O3 C8H14O3 C9H16O3 C10H18O3 C11H20O3 C12H22O3 C13H24O3 C14H26O3 C15H28O3 C16H30O3 C17H32O3 C18H34O3
2 2 2 2 2 2 2 2 2 2 2 2 2 2
115.04007 129.05572 143.07137 157.08702 171.10267 185.11832 199.13397 213.14962 227.16527 241.18092 255.19657 269.21222 283.22787 297.24352
115.03996 129.05562 143.07132 157.08694 171.10256 185.11817 199.13380 213.14939 227.16505 241.18070 255.19640 269.21192 283.22752 297.24315
-0.94 -0.79 -0.36 -0.51 -0.66 -0.79 -0.85 -1.07 -0.96 -0.91 -0.66 -1.11 -1.25 -1.24
5 6 7 8 9 10 11 12 13 14 15 16 17 18
C5H10O3 C6H12O3 C7H14O3 C8H16O3 C9H18O3 C10H20O3 C11H22O3 C12H24O3 C13H26O3 C14H28O3 C15H30O3 C16H32O3 C17H34O3 C18H36O3
1 1 1 1 1 1 1 1 1 1 1 1 1 1
117.05572 131.07137 145.08702 159.10267 173.11832 187.13397 201.14962 215.16527 229.18092 243.19657 257.21222 271.22787 285.24352 299.25917
117.05567 131.07130 145.08696 159.10262 173.11818 187.13383 201.14946 215.16506 229.18072 243.19628 257.21194 271.22763 285.24322 299.25891
-0.41 -0.52 -0.44 -0.30 -0.80 -0.74 -0.81 -0.97 -0.87 -1.21 -1.08 -0.88 -1.05 -0.86
3 4 5 6 7 8 9 10 11 12 13 14 15 16 18 19 20 22
C3H4O4 C4H6O4 C5H8O4 C6H10O4 C7H12O4 C8H14O4 C9H16O4 C10H18O4 C11H20O4 C12H22O4 C13H24O4 C14H26O4 C15H28O4 C16H30O4 C18H34O4 C19H36O4 C20H38O4 C22H42O4
2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2
103.00368 117.01933 131.03498 145.05063 159.06628 173.08193 187.09758 201.11323 215.12888 229.14453 243.16018 257.17583 271.19148 285.20713 313.23843 327.25408 341.26973 369.30103
103.00356 117.01922 131.03489 145.05054 159.06616 173.08178 187.09740 201.11306 215.12867 229.14431 243.15997 257.17562 271.19149 285.20684 313.23808 327.25368 341.26922 369.30059
-1.17 -0.98 -0.71 -0.66 -0.75 -0.90 -0.99 -0.88 -0.99 -0.97 -0.88 -0.83 0.02 -1.03 -1.12 -1.23 -1.52 -1.20
3
C3H8O4S
0
139.00705
139.00696
-0.68
Saturated Hydroxy-Fatty Acids hydroxy-pentanoic acid hydroxy-hexanoic acid hydroxy-heptanoic acid hydroxy-octanoic acid hydroxy-nonanoic acid hydroxy-decanoic acid hydroxy-undecanoic acid hydroxy-dodecanoic acid hydroxy-tridecanoic acid hydroxy-tetradecanoic acid hydroxy-pentadecanoic acid hydroxy-hexadecanoic acid hydroxy-heptadecanoic acid hydroxy-octadecanoic acid Saturated Dicarboxylic Acidsc propanedioic acid butanedioic acid pentanedioic acid hexanedioic acid heptanedioic acid octanedioic acid nonanedioic acid decanedioic acid undecanedioic acid dodecanedioic acid tridecanedioic acid tetradecanedioic acid pentadecanedioic acid hexadecanedioic acid octadecanedioic acid nonadecanedioic acid eicosanedioic acid doeicosanedioic acid Sulfates monopropylsulfate
S11
monotetrasulfate monopentasulfate monohexylsulfate monoheptasulfate monooctylsulfate monononasulfate monodecylsulfate monoundecylsulfate monododecylsulfate monotridecylsulfate monotetradecylsulfate monopentadecylsulfate monohexadecylsulfate monoheptadecylsulfate
4 5 6 7 8 9 10 11 12 13 14 15 16 17
C4H10O4S C5H12O4S C6H14O4S C7H16O4S C8H18O4S C9H20O4S C10H22O4S C11H24O4S C12H26O4S C13H28O4S C14H30O4S C15H32O4S C16H34O4S C17H36O4S
0 0 0 0 0 0 0 0 0 0 0 0 0 0
153.02270 167.03835 181.05400 195.06965 209.08530 223.10095 237.11660 251.13225 265.14790 279.16355 293.17920 307.19485 321.21050 335.22615
153.02258 167.03821 181.05384 195.06945 209.08531 223.10075 237.11628 251.13202 265.14756 279.16319 293.17879 307.19445 321.21007 335.22567
-0.81 -0.87 -0.91 -1.05 0.03 -0.92 -1.39 -0.95 -1.30 -1.31 -1.41 -1.32 -1.35 -1.45
16 17 18 19 20
C16H26O3S C17H28O3S C18H30O3S C19H32O3S C20H34O3S
4 4 4 4 4
297.15299 311.16864 325.18429 339.19994 353.21559
297.15257 311.16816 325.18374 339.19936 353.21501
-1.41 -1.54 -1.69 -1.71 -1.64
Linear Alkylbenzene Sulfonatesd decylbenzenesulfonate undecylbenzenesulfonate dodecylbenzenesulfonate tridecylbenzenesulfonate tetradecylbenzenesulfonate a
The empirical formulas for the proposed linear chain acids may also correspond to branched chain acids as well.
b
All monoisotopic masses shown correspond to the deprotonated ion. Empirical formulas for the tentatively identified dicarboxylic acids may also correspond to saturated oxo-hydroxycarboxylic acids. d Stereo-connectivity are not shown as they commonly exist as mixtures of species with the benzenesulfonate at various positions of the alkyl chain. e The measured masses shown were obtained from the analysis of fine SSA collected during time period 3. c
S12
Table S3. Relative intensities of fatty acids and fatty acid derivatives tentatively identified during the mesocosm experiment. Values were calculated based on the signal intensities of the [M-H]- ion for each compound in composite spectra from UPLC-ESI-QToF MS analysis. Acid Class
Acid Name (Tentative)
Saturated Fatty Acids octanoic acid nonanoic acid decanoic acid undecanoic acid dodecanoic acid tridecanoic acid tetradecanoic acid pentadecanoic acid hexadecanoic acid heptadecanoic acid octadecanoic acid nonadecanoic acid eicosanoic acid uneicosanoic acid dodeicosanoic acid trieicosanoic acid tetracosanoic acid
Carbon #
Degree of Unsat.
SSW
8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
0.2 1 0.7 0.2 1 0.4 4 2 34 3 39 0.1 0.1 0.04 0.1 NRa 0.05
1 2 3 0.9 3 1.3 5 4 25 3 28 0.1 0.7 2 2 0.1 0.2
0.2 0.3 0.3 0.2 2 1.1 6 3 32 3 38 0.3 0.2 0.2 0.1 0.03 0.1
0.2 0.6 0.6 0.2 2 1.9 8 4 32 3 30 0.2 0.1 0.04 0.1 0.02 NR
86
81
86
84
0.01 0.02 0.05 0.05 0.4 0.1 1 0.09 0.03 0.02 0.03 0.02 0.01 0.3 0.1 0.01 0.03 0.05 0.06 0.04 0.02 0.05 0.01 0.03 0.1 0.04
0.03 0.02 0.1 0.1 1 0.3 0.8 0.2 0.05 0.03 0.01 0.3 0.03 2 0.5 0.02 0.01 0.01 NR 0.02 0.01 0.05 NR 0.02 0.3 NR
0.03 0.04 0.07 0.09 0.3 0.2 0.9 0.02 0.02 0.03 0.03 0.06 0.1 0.5 0.1 0.01 0.1 0.04 0.04 0.02 0.03 0.1 0.1 0.07 0.2 0.1
0.04 0.05 0.1 0.2 0.4 0.2 0.6 0.1 0.05 0.05 0.1 0.1 0.07 0.2 0.1 0.02 0.06 0.06 0.07 0.04 0.01 0.04 0.03 0.02 0.09 0.02
Sum of Saturated Fatty Acids Unsaturated Fatty Acids dodecenoic acid tridecenoic acid tetradecenoic acid pentadecenoic acid hexadecenoic acid heptadecenoic acid octadecenoic acid dodecadienoic acid tridecadienoic acid tetradecadienoic acid pentadecadienoic acid hexadecadienoic acid heptadecadienoic acid octaadecadienoic acid tetradecatrienoic acid hexadecatrienoic acid octadecatrienoic acid undecatetraenoic acid dodecatetraenoic acid hexadecatetraenoic acid octadecatetraenoic acid eicosatetraenoic acid uneicosatetraenoic acid octadecapentaenoic acid eicosapentaenoic acid doeicosapentaenoic acid
12 13 14 15 16 17 18 12 13 14 15 16 17 18 14 16 18 11 12 16 18 20 21 18 20 22
1 1 1 1 1 1 1 2 2 2 2 2 2 2 3 3 3 4 4 4 4 4 4 5 5 5
S13
Percent of Total FA Signal SSML Coarse SSA
Fine SSA
doeicosahexaenoic acid
22
6
Sum of Unsaturated Fatty Acids Oxygenated Saturated Fatty Acids oxopentanoic acid oxohexanoic acid oxoheptanoic acid oxooctanoic acid oxononanoic acid oxodecanoic acid oxoundecanoic acid oxododecanoic acid oxotridecanoic acid oxotetradecanoic acid oxopentadecanoic acid oxohexadecanoic acid oxoheptadecanoic acid oxooctadecanoic acid
5 6 7 8 9 10 11 12 13 14 15 16 17 18
0 0 0 0 0 0 0 0 0 0 0 0 0 0
Sum of Saturated Oxo-Fatty Acids Hydroxylated Saturated Fatty Acids hydroxy-pentanoic acid hydroxy-hexanoic acid hydroxy-heptanoic acid hydroxy-octanoic acid hydroxy-nonanoic acid hydroxy-decanoic acid hydroxy-undecanoic acid hydroxy-dodecanoic acid hydroxy-tridecanoic acid hydroxy-tetradecanoic acid hydroxy-pentadecanoic acid hydroxy-hexadecanoic acid hydroxy-heptadecanoic acid hydroxy-octadecanoic acid
5 6 7 8 9 10 11 12 13 14 15 16 17 18
0 0 0 0 0 0 0 0 0 0 0 0 0 0
Sum of Saturated Oxo-Fatty Acids a
0.02
0.02
0.07
0.04
3
6
4
3
1 0.1 0.0 0.03 0.3 0.05 0.05 0.06 0.03 0.04 0.1 0.23 0.01 0.4
0.09 0.1 0.1 0.02 0.1 0.1 0.08 0.08 0.04 0.08 0.1 0.25 0.01 0.6
0.2 0.1 0.06 0.1 0.3 0.1 0.09 0.09 0.06 0.07 0.06 0.2 0.03 0.4
0.8 0.1 0.09 0.2 0.5 0.3 0.2 0.2 0.1 0.1 0.2 0.3 0.04 0.4
3
2
2
4
0.02 0.09 0.1 0.07 0.1 0.0 0.03 0.1 0.06 0.05 NR 0.2 0.003 NR
0.1 0.07 0.08 0.08 0.08 0.07 0.05 0.0 0.07 0.09 0.01 0.3 0.03 0.2
0.03 0.02 0.02 0.06 0.04 0.03 0.03 0.04 0.01 0.02 0.01 0.1 0.03 0.1
0.04 0.04 0.04 0.09 0.1 0.08 0.05 0.08 0.004 0.1 0.01 0.2 0.02 0.2
0.8
1
0.6
1
"NR" denotes the HRMS response in the samples was below that observed in the field blanks.
S14
