Macroporous resins (Amberlite® XAD4, Amberlite® XAD7HP, and Supelite™ DAX-8) were purchased from Sigma (Aldrich, Germany). The physical properties of these resins are summarized in Table 5 . To remove monomers and porogenic agents trapped inside the pores, the resins were soaked in 95% ethanol at a resin/solvent ratio of 1:20 for 24 h. The resins were then washed with deionized water and soaked again in 1 M sodium hydroxide for another 24 h after several washes with deionized water; then, the resins were soaked in 1 M hydrochloric acid for 24 h and washed again with deionized water. The resins were dried (60 °C) in a universal drying oven (model 100–800, Memmert, Schwabach, Germany) until constant weight was observed.
Supelite dax 8
Manufactured by Merck Group
Sourced in Germany
Supelite™ DAX-8 is a high-performance liquid chromatography (HPLC) sorbent designed for the purification and separation of a wide range of organic compounds. It features a bonded octyl (C8) stationary phase that provides excellent retention and selectivity for a variety of analytes. The sorbent's high surface area and uniform particle size distribution contribute to efficient chromatographic separations.
Automatically generated - may contain errors
Lab products found in correlation
Amberlite xad4, by Merck Group (2 mentions)
Amberlite xad7hp, by Merck Group (2 mentions)
Amberlite ira958, by Merck Group (1 mentions)
Diaion hp 20, by Merck Group (1 mentions)
Amberlite xad 2, by Merck Group (1 mentions)
Novozym 435, by Novozymes (1 mentions)
Lipozyme tl im, by Novozymes (1 mentions)
Lecitase ultra, by Novozymes (1 mentions)
Sartocollect 1, by Sartorius (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Polyvinylpyrrolidone, by Merck Group (1 mentions)
Oca20, by Dataphysics (1 mentions)
Quercetin, by Thermo Fisher Scientific (1 mentions)
Rutin, by Thermo Fisher Scientific (1 mentions)
Divinylbenzene, by Thermo Fisher Scientific (1 mentions)
Millipore water, by Merck Group (1 mentions)
Styrene, by Merck Group (1 mentions)
2 2 azobis 2 methylpropionitrile, by Merck Group (1 mentions)
Methanol meoh, by Thermo Fisher Scientific (1 mentions)
Gallic acid, by Merck Group (1 mentions)
6 protocols using supelite dax 8
1
Preparation of Macroporous Resins
2
Fractionation of Organic Matter using Resin Chromatography
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Extract was fractionated by using different resins (Supelite DAX-8, Amberlite XAD-4 and Amberlite IRA-958, Sigma-Aldrich, St. Louis, MO, USA) [41 (link),42 (link)]. The fractionation approach suggested by Tao et al. [16 ] was used and is shown schematically in Figure 2 . Each extracts was adjusted to pH = 2 and fed onto a Supelite DAX-8 non-functionalized resin, which retained strongly hydrophobic (SH) organic matter. This fraction was eluted with 0.1 mol/L NaOH. The unabsorbed concentrate from the DAX-8 resin was fed onto an Amberlite XAD-4 resin, which retained weakly hydrophobic (WH) organic matter. This fraction was also eluted with 0.1 mol/L NaOH. The unabsorbed concentrate from the XAD-4 resin, which comprised hydrophilic organic matter attributed to proteins, amino acids, and carbohydrates, was fed onto an Amberlite IRA-958 anion exchange resin, which retained charged hydrophilic (CH) material. This fraction was eluted with 1 mol/L NaOH/NaCl. The remaining neutral hydrophilic (NH) material was not retained by any of the resins.
3
Lipid Modification Protocol: Fatty Acid Profiling
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Analytical standards Supelco 37-component fatty acid methyl ester (FAME mix), methyl heptadecanoate (C17:0), and methyl cis-15-tetracosenoate (C24:1) were purchased from Sigma-Aldrich (Castle Hill, Australia). Granulated soybean L-α-phosphatidylcholine PC95 (PC, purity > 95%) was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Lipases, including Lipozyme TL IM (from Thermomyces lanuginose, immobilized on silica gel carrier), Novozym 435 (from Candida antartica, immobilized on a microporous acrylic resin via interfacial activation), and Phospholipase A1 (Lecitase® Ultra, from T. lanuginose/Fusarium oxysporum) were donated from Novozymes A/S (Bagsværd, Denmark). Amberlite XAD-7 HP, Amberlite XAD-2, Supelite DAX-8, and Diaion HP-20 were purchased from Sigma–Aldrich, Inc. (Castle Hill, Australia). Nervonic acid (98% purity) was purchased from Shenzhen Dieckmann Technology Development Co., Ltd. (Shenzhen, China). Chromatographic grade n-hexane, chloroform, and methanol were obtained from ECP Limited, Auckland, New Zealand.
4
Characterization of Polymeric Beads
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The following polymeric beads were purchased and used in this study: cellulose from Perloza S.r.o (Czech Republic); Sepharose agarose, sephacryl, dextran from GE Healthcare (Germany); Fastline agarose from DSM biologics (The Netherlands); polyacrylamide, ceramic hydroxyapatite I and II, ceramic flouroapatite I and II from Bio-Rad (Germany); polyvinylpyrrolidone, Supelite™ DAX-8, poly(styrene-co-divinylbenzene) from Sigma-Aldrich (Germany); poly(phenol–formaldehyde), poly(ethyl methacrylate) from Polysciences (Germany); Toyopearl HW 65 from TOSOH Bioscience (Germany); and geniaLab (Germany) provided beads of agar, alginate and chitosan. Carrageenan was obtained from Carl Roth (Germany). α-Bromonaphthalene (99% purity) and formamide (99.5% purity) were obtained from Fluka, Buchs, Switzerland. Water was ultrapure quality. All other chemicals used in the buffer preparation were of analytical grade and obtained from Applichem (Germany). The goniometric system (OCA 20) was obtained from Data- Physics Instruments (Germany). Zeta potential measurements were measured using a ZetaSizer Nano ZS from Malvern Instruments (UK). A weighing balance CPA423S connected to computer via RS-232 serial connector and Sartocollect 1.0 was used to control and record the data was obtained from Sartorius AG (Germany).
5
Supelite DAX-8 Resin Synthesis
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Quercetin (hydrate, purity 95%) and rutin (purity 97%) were supplied by Acros Organics. Gallic acid (assay 97.5-102.5%, titration), styrene (S, 99% purity), ethylene glycol dimethacrylate (EGDMA, 98% purity), 2,2′-Azobis(2-methylpropionitrile) (AIBN, 98% purity) and Supelite™ DAX-8 (an adsorbent of moderate polarity also referred as polymethylmethacrylate resin) were purchased from Sigma Aldrich. Divinylbenzene (DVB, 80% purity) and 4-vinylpyridine (4VP, 95% purity) were provided by Alfa Aesar. Analytical reagent grades for acetonitrile (ACN), dimethylformamide (DMF), acetic acid (AcOH), and methanol (MeOH) were bought from Fisher Scientific and for ethanol (EtOH) from PanReac. Millipore water (Milli-Q quality) was used in all the experiments unless otherwise mentioned.
6
Groundwater Humic Substances Extraction
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For all experiments, Milli-Q grade pure water and analyticalgrade chemicals purchased from Wako Pure Chemical Industries were used, unless otherwise noted. HA and FA were extracted from groundwater collected at the À250 m gallery of the Horonobe URL located in the northern part of Hokkaido Prefecture, Japan. The geology and geochemistry of the site are described elsewhere. 35, 42 The À250 m gallery is located at the boundary of the Pliocene Koetoi and the Miocene Wakkanai formations, which are composed of diatomaceous and siliceous mudstones, respectively. Groundwater at the sampling location is a weakly alkaline Na + /HCO 3 À type with relatively high total organic carbon (TOC) and Cl À levels. Groundwater aer ltration and acidication was passed through a column packed with DAX-8 resins (Supelite DAX-8, Sigma-Aldrich). Separation and extraction of HA and FA fractions from the loaded resins and subsequent purication was performed in a laboratory on the surface, according to the protocol recommended by the International Humic Substances Society (IHSS). 43 In total 6.6 g of HA and 3.5 g of FA were obtained by treating approximately 6000 L of the groundwater. These values correspond to approximately 8.5 and 4.5% of the TOC in the groundwater, respectively. Hereaer, the HA and FA fractions are denoted as HHA and HFA, respectively.
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