Supporting Information Appendix - eScholarship

Photochemical Production of Polyols arising from Significant Photo-transformation of Dissolved Organic Matter in the Oligotrophic Surface Ocean Michael Gonsior a,b*, Norbert Hertkorn c, Maureen H. Conte d, William J. Cooper e , David Bastviken b , Ellen Druffel f and Philippe Schmitt-Kopplin c,g a

University of Maryland Center for Environmental Science, Chesapeake Biological Laboratory, Solomons, USA b

Linköping University, Department of Thematic Studies, UnitWater and Environment, Linköping, Sweden c

Helmholtz Center Munich, German Research Center for Environmental Health, Neuherberg, Germany d

Ecosystems Center, Marine Biological Laboratory, Woods Hole, USA

e

Urban Water Research Center, Department of Environmental and Civil Engineering, University of California, Irvine, USA f

Department of Earth System Science, University of California, Irvine, USA

g

Department for Chemical-Technical Analysis, Research Center Weihenstephan for Brewing and Food Quality, Technische Universität München, D-85354 Freising-Weihenstephan, Germany *

Corresponding author. phone: +14103267245, Email address: [email protected]

Supporting Information

Table S1: Acquisition parameters of NMR spectra of SPE-DOM (ATL: Atlantic Ocean SPEDOM, PAC: Pacific Ocean SPE-DOM, ATLPAC after: consolidated ATL+PAC after photoirradiation), shown according to figures. NS: number of scans (for 2D NMR: F2); AQ: acquisition time [ms]; D1: relaxation delay [ms]; NE: number of F1 increments in 2D NMR spectra; WDW1, WDW2: apodization functions in F1/ F2 (EM/GM: line broadening factor [Hz]; QS: shifted square sine bell; SI: sine bell); PR1, PR2: coefficients used for windowing functions WDW1, WDW2, EM/GM are given in [Hz], SI/QS derived functions indicate shift by π/n. Total NMR acquisition time AQΣ is computed as follows: AQΣ = NS × (D1 + AQ) × NE, with NE = 1 for 1D NMR spectra. spectrum 1 H NMR 1 H NMR 1 H NMR 1 H NMR 1 H NMR

SPE-DOM ATL before ATL after PAC before PAC after ATLPAC after

NS 7024 8192 1024 2560 1024

AQ [ms] 5000 5000 5000 5000 5000

D1 [ms] 5000 5000 5000 5000 5000

NE -

WDW1 -

WDW2 EM EM EM EM EM

PR1 -

PR2 1 1 1 1 1

H,1H JRES

ATL before

400

750

750

65

QS

QS

0

0

ATLPAC after

192

750

750

849

QS

EM

3

2

ATLPAC after

1600

250

1250

128

QS

EM

2.5

10

ATLPAC after

1600

250

1250

90

QS

EM

2.5

10

ATLPAC after

1600

250

1250

356

QS

EM

2

2

PAC after ATL after

32 32

250 250

1250 1250

1408 1535

SI SI

EM EM

2.5 2.5

7.5 7.5

1 1 1

1

H, H COSY 13

H, C DEPTHSQC (CH3) 1 13 H, C DEPTHSQC (CH2) 1

1 1

13

H, C HSQCTOCSY 1

H, H TOCSY H,1H TOCSY

Figure S1a: Ultraviolet-visible (UV-Vis) spectra of Pacific Ocean and Atlantic Ocean SPEDOM before and after the exposure to 24 h of simulated sunlight.

Figure S1b: Excitation emission matrix fluorescence (EEM) of Pacific Ocean (aa) and Atlantic Ocean (ba) SPE-DOM and the associated relative decrease in fluorescence (ab, bb) after the exposure to 24 h of simulated sunlight.

a

b

c

Figure S2: Consolidated negative electrospray FT-MS results of the photochemically produced/refractory marine DOM, (a) van Krevelen, (b) H/C ratios and (c) O/C ratios versus the exact mass in Dalton. Note: size of bubbles represent relative change in intensity; orange: photodegraded (decrease in intensity) and green: photo-produced (increase in intensity).

a

b

c

d

Figure S3: The KMD-z* Kendrick mass diagram: a recently introduced visualization tool for ultrahigh resolution mass spectrometry++. (a): KMD-Z mass diagram of all assigned SPE-DOM formulas, prior to photolysis; (b) KMD-Z mass diagram between 307-362 Da to visualize the three most important spacings of molecular formulas within NOM in one diagram (SPE-DOM prior to photolysis); (c) KMD-Z mass diagram of photochemically-induced mass peaks which were increased in amplitude following photolysis (d) decreased intensities of SPE-DOM mass peaks. Note: The size of the bubbles in (a,b) refers to the rel. abundance of the mass peaks and in (c, d) to the rel. change in intensity; orange: photo-degraded (decrease in intensity) and green: photo-produced (increase in intensity). Note: ++ adapted from: Shakeri Yekta, S.; Gonsior, M.; Schmitt-Kopplin, P.; Svensson, B. H., Characterization of Dissolved Organic Matter in Full Scale Continuous Stirred Tank Biogas Reactors Using Ultrahigh Resolution Mass Spectrometry: A Qualitative Overview. Environ. Sci. Technol. 2012, 46, (22), 12711-12719.

Figure S4: 1H NMR spectra (500 MHz) of the Atlantic and Pacific SPE-DOM in CD3OD before and after the exposure to 24h solar simulated irradiation.

Figure S5: 1H, 13C DEPT HSQC NMR spectra of consolidated Atlantic and Pacific SPE-DOM after photo irradiation. Top: CH2-selective (CH2: green, CH and CH3: gray); bottom: CH3selective (CH3: red, CH and CH2: gray), with key substructures indicated; orange: 1H NMR chemical shift range (δH < 1.0 ppm) of methyl terminating classical aliphatic chains (H3CCCC); a sizable fraction of C-CH3 cross peaks (δH > 1.0 ppm) indicated proximity of methyl and carboxylic groups like in carboxyl-rich alicyclic compounds (Hertkorn et al., 2006; Hertkorn et al., 2013). Shaded boxes: green, chemical shift range of methylene in polyols (and common carbohydrates; cf. Fig. S7); purple: chemical shift range of methine in polyols (cf. Fig. S7).

Hertkorn, N. et al., 2006. Characterization of a major refractory component of marine dissolved organic matter. Geochim. Cosmochim. Acta, 70(12): 2990-3010. Hertkorn, N., Harir, M., Koch, B.P., Michalke, B. and Schmitt-Kopplin, P., 2013. High-field NMR spectroscopy and FTICR mass spectrometry: powerful discovery tools for the molecular level characterization of marine dissolved organic matter. Biogeosciences, 10(3): 1583-1624.

Figure S6: JRES NMR spectra (B0 = 11.7 T, left and 18.8 T right) of Atlantic SPE-DOM before (right) and after (left) photo irradiation. Near featureless NMR spectrum of pre-photo SPE-DOM indicated a rather even abundance distribution of many DOM constituents (asterisk: 13C satellite of CD3OD). JRES cross peaks highlighted in purple indicate C*-CH2OH spin systems with asymmetrically substituted carbon, typical of polyols.

Figure S7: NMR spectra (B0 = 11.7 T) of common polyols (C3: glycerol, C4: erythrol, C5: arabitol, C6: sorbitol) in CD3OD, A: JRES NMR; B: COSY NMR; C: DEPT-HSQC NMR spectra, with methylene (green) and methane (blue) indicated, including common chemical shift windows (green and purple shaded boxes; cf. Fig. S5).

Figure S8: 1H NMR spectra (500 MHz) referring to second solid phase extraction (PPL) of consolidated Atlantic and Pacific SPE-DOM to assess extraction efficiency of photochemically formed polyols (Fig. 2). (A) methanolic sample solution (CD3OD) after one year storage at -20 o C (demonstrates full sample integrity without decomposition; blue shade: chemical shift range of JRES NMR spectrum, Fig. S6). (B) consolidated Atlantic and Pacific SPE-DOM in D2O after solvent exchange (shows complete dissolution of polyol fraction of SPE-DOM with “background” SPE-DOM somewhat attenuated and displacements of polyol chemical shift effected by solvent change). (C) Extracted photo-irradiated SPE-DOM in CD3OD indicates largely the absence of polyols. (D) direct eluate from initial loading of PPL cartridge, reconstituted in CD3OD, indicates presence of polyols and other protons at oxygenated aliphatic positions: OCH, suggesting inferior extraction efficiency of the polyol fraction to the PPL resin.