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1 Supporting Information Proposed guidelines for solid phase extraction of Suwannee River dissolved organic matter Yan Li, Mourad Harir,, Marianna Lucio, Basem Kanawati, Kirill Smirnov, Ruth Flerus, Boris P. Koch,, Philippe Schmitt-Kopplin,, Norbert Hertkorn,* Helmholtz Zentrum München, German Research Center for Environmental Health, Research Unit Analytical Biogeochemistry (BGC), Ingolstädter Landstrasse 1, Neuherberg, Germany Chair of Analytical Food Chemistry, Technische Universität München, Freising, Germany GEOMAR Helmholtz-Zentrum für Ozeanforschung Kiel, Düsternbrooker Weg 20, Kiel, Germany Alfred Wegener Institute for Polar und Marine Research, Am Handelshafen 12, Bremerhaven, Germany Hochschule Bremerhaven, University of Applied Sciences, An der Karlstadt 8, Bremerhaven; Germany * Corresponding author contact: Phone: Fax: hertkorn@helmholtz-muenchen.de Number of figures: 10 Number of tables: 1 S-1
2 Figure S1. Van Krevelen diagrams of (A) permeates and (B) wash liquids obtained at different loading volumes at DOC concentrations of 80 mg/l and a flow rate of 0.5 ml/min on 100 mg PPL cartridges; mass-edited H/C ratios of (C) permeates and (D) wash liquids. The pie charts indicate the count of assigned formulas with color code provided. The bubble area corresponds to the mass peak intensities observed in FTICR mass spectra. S-2
3 Figure S2. 1 H NMR spectra (500 MHz, CD 3 OD) of permeates obtained at the following conditions: (A) different loading volumes at a DOC concentration of 80 mg/l and a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (B) variable DOC concentration with identical total loading mass of 0.5 mg DOC on 100 mg PPL cartridges; (C) different DOC concentrations at identical loading volume of 6.25 ml and a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (D) different flow rates at a loading volume of 6.25 ml and DOC concentration of 20 mg/l (identical DOC/PPL mass ratio 1:800) on 100 mg PPL cartridges; and (E) up-scaling with different sizes of PPL cartridges at identical DOC/PPL mass ratio (1:800) at 0.5 ml/min. S-3
4 Figure S3. 1 H NMR spectra (500 MHz, CD 3 OD) of wash liquids obtained at the following conditions: (A) different loading volumes at a DOC concentration of 80 mg/l and a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (B) variable DOC concentration with identical total loading mass of 0.5 mg DOC on 100 mg PPL cartridges; (C) different DOC concentrations at a loading volume of 6.25 ml and a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (D) different flow rates at a loading volume of 6.25 ml and DOC concentration of 20 mg/l (DOC/PPL mass ratio of 1:800) on 100 mg PPL cartridges; and (E) up-scaling with different sizes of PPL cartridges at identical DOC/PPL mass ratio of 1:800 at 0.5 ml/min. S-4
5 Figure S4. DOC concentrations and recoveries under different SPE conditions. (A) Identical total DOC mass at loading DOC mass of 0.5 mg with variable concentrations and volumes at a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (B) variable flow rates at 20 mg/l DOC concentration with 6.25 ml loading volume (DOC/PPL mass ratio of 1:800) on 100 mg PPL cartridges; and (C) up-scaling with PPL cartridge size at a DOC/PPL mass ratio of 1:800 at a flow rate of 0.5 ml/min. The DOC values of the permeates, wash liquids and eluates are shown on the left y-axis, and the DOC recovery is presented on the right y-axis. The recovery was calculated according to the formula: 100% (DOC of eluate)/(original DOC). S-5
6 Figure S5. Van Krevelen diagrams (upper panel) and mass-edited H/C ratios of common masses in FTICR mass spectra of all eluates (lower panel) obtained under different conditions: (A) DOC mass of 0.5 mg at a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (B) different flow rates with a DOC concentration of 20 mg/l and a loading volume of 6.25 ml (DOC/PPL mass ratio of 1:800) on 100 mg PPL cartridges; (C) different up-scaling with PPL cartridges of different sizes at a DOC/PPL mass ratio of 1:800 and a flow rate of 0.5 ml/min. The bubble areas correspond to the FTICR mass peak intensities. S-6
7 Figure S6. 1 H NMR spectra (500 MHz, CD 3 OD) of eluates obtained under different conditions: (A) DOC mass of 0.5 mg at a flow rate of 0.5 ml/min on 100 mg PPL cartridges; (B) different flow rates with a DOC concentration of 20 mg/l and a loading volume of 6.25 ml (DOC/PPL mass ratio of 1:800) on 100 mg PPL cartridges; (C) different up-scaling with PPL cartridges of different sizes at a DOC/PPL mass ratio of 1:800 and a flow rate of 0.5 ml/min. S-7
8 Figure S7. 1 H NMR spectra (800 MHz, CD 3 OD/D 2 O: 50/50 v/v; asterisk: HD 2 OD) of (lower panel) Suwannee river organic matter isolates [blue: Suwannee River water (Suw-water); brown: IHSS reverse osmosis NOM (2R101N: Suw-RO); black/gray: SPE/PPL eluate], and the corresponding difference NMR spectra (top panel, with excess relative abundance colored according to the parent NMR spectra). The shaded box indicates potential carbohydrate NMR resonances (cf. text). S-8
9 Figure S8. 1 H NMR section integrals for key substructures of the three Suwannee River organic matter isolates (Suwannee River water (Suw-water); IHSS reverse osmosis NOM (2R101N; Suw-RO); and SPE/PPL eluates) and a cluster dendrogram of the three samples obtained from 1 H NMR section integrals (0.01 ppm resolution). Figure S9. FTMS-derived Venn diagram of the number of assigned formulas of the three Suwannee River organic matter isolates (Suwannee River water (Suw-water); IHSS reverse osmosis NOM (2R101N; Suw-RO), and SPE/PPL eluate). S-9
10 members of molecular series SPE/PPL Suw-water Suw-RO Suw-RO* CHO-compounds CHOS-compounds CHNO-compounds CHNOS-compounds Total number of assigned mass peaks average H [%] average C [%] average O [%] average N [%] average S [%] computed average H/C ratio computed average O/C ratio computed average C/N ratio computed average C/S ratio DBE average DBE/C average mass weighted average Table S1. Counts of mass peaks as computed from FTICR MS data for singly charged ions with nitrogen rule check and 500 ppb tolerance. Suw-RO*: elemental compositions in terms of atomic ratios as computed from the IHSS database ( S-10
11 Figure S10. Van Krevelen diagrams of the assigned formulas detected in the three samples (left column: from top to bottom: Suwannee River water (Suw-water); IHSS reverse osmosis NOM (Suw-RO); and SPE/PPL eluate). Mass-edited H/C ratios of assigned molecular formulas (right column). The bubble areas correspond to the FTICR mass peak intensities; FTMS-derived cluster analysis shown on the right margin. S-11
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