Milli q 18 mω

In the experiment, the sodium sulfate, ammonium sulfate, and sodium hydroxide purchased from the Sinopharm Chemical Reagent Co., Ltd. (Beijing, China) were used in the chemical analyses. Deionized water (Millipore Milli-Q 18 MΩ, Bedford, MA, USA) was used throughout. The Selemion homogeneous cation exchange membrane CMV and Selemion anion exchange membrane AMV used during the experiment were purchased from Asahi Glass Co., Suzhou, China and the bipolar membrane (BPM) was provided by Tokuyama Astom Co., Tokuyama, Japan. The main properties of the ion exchange membrane are presented in Table 1.

High oleic soybean oil (Perdue Agribusiness LLC, Salisbury, MD, US), sodium dodecyl sulfate (SDS, VWR; Solon, OH, US), and sodium chloride (VWR; Solon, OH, US) were used as received. Methyl methacrylate (MMA, Alfa-Aesar; Ward Hill, MA, USA), butyl acrylate (BA, TCI America, Portland, OR, USA), styrene (St, Sigma-Aldrich, St. Louis, MO, US), and acrylic acid (AA, Alfa-Aesar; Ward Hill, MA, USA) were distilled under vacuum to remove the inhibitor and stored in a refrigerator. Azobisisobutyronitrile (AIBN; Sigma-Aldrich, St. Louis, MO, USA) was purified with recrystallization from methanol. All other solvents used were reagent grade or better and used as received. Deionized water was used throughout the study (Millipore water, MilliQ, 18 MΩ).

Chemicals and solvents were all purchased from Sigma-Aldrich and ultra-pure water was used for buffer preparation (Milli-Q 18 MΩ, Millipore, France).

The DNA samples used in this study were obtained from Sangon Biotech (Shanghai, China) for experimental purposes; the DNA sequences and modifications are listed in Table 1. All DNA samples were purified by HPLC, stocked in 1× TE buffer (pH = 7.8), and stored at −20 °C.
T4 DNA ligase and phi29 DNA polymerase were purchased from Beyotime Biotech (Shanghai, China). Beyotime Biotech also provided a 10× T4 DNA ligation buffer and 10× phi29 DNA polymerase reaction buffer. Streptavidin immunomagnetic beads (IMBs, 10 mg/mL, 0.5 μm), dNTPs, and BSA were purchased from Sangon Biotech (Shanghai, China). Hemin, 3, 3′, 5, 5′-tetramethylbenzidine (TMB), and hydrogen peroxide (H₂O₂) were purchased from Beyotime Biotech. All other chemical reagents were bought from Sangon Biotech and were of analytical grade. All solutions in this study were prepared using ultrapure water sourced from Milli-Q (18 MΩ, Merck Millipore, Shanghai, China). The buffers used in this method are listed as follows:
Binding Buffer (10×): 100 mM Tris-HCl, 500 mM MgCl₂, 500 mM NaCl.
Enzyme Activity Buffer: 100 mM Tris Base, 120 mM NaCl, 10 mM MgCl₂, 100 mM KCl.
Substrate Buffer: 200 mM Na₂HPO₄, 100 mM citric acid.

The polyphenols in the selected fractions were identified and quantified using Waters Acquity (Waters Corporation, Milford, MA, USA) ultra-high performance liquid chromatography coupled with tandem quadrupole mass spectrometry (UHPLC-TQD-MS). The LC separation of the analytes was performed on a Waters Acquity HSS T3 UHPLC column (1.8 μm, 2.1 × 100 mm) using milli-Q^® (18 mΩ) (Merck Millipore, Molsheim, France) water (mobile phase A) and acetonitrile:methanol (1:1) containing 0.5% formic acid (mobile phase B). A gradient program of 0–2.5 min 2% B, 2.5–3 min 10% B, 3–7.5 min 15% B, 7.5–8.5 min 35% B, 8.5–9.5 min 98% B, and 9.5–10.0 min 2% B at a flow rate of 0.5 mL/min was used. A multiple reaction monitoring (MRM) approach was taken for the mass spectrometric determination of the polyphenols using argon as collision gas. The parameters for MRM transitions were obtained using the Waters integrated IntellistartTM software (Waters Corp., Milford, MA, USA) (Table 2).

The polyphenols in the selected fractions were identified and quantified using Waters Acquity (Waters Corporation, Milford, MA, USA) ultra-high performance liquid chromatography coupled with tandem quadrupole mass spectrometry (UHPLC-TQD-MS). The LC separation of the analytes was performed on a Waters Acquity HSS T3 UHPLC column (1.8 μm, 2.1 × 100 mm) using milli-Q^® (18 mΩ) (Merck Millipore, Molsheim, France) water (mobile phase A) and acetonitrile:methanol (1:1) containing 0.5% formic acid (mobile phase B). A gradient program of 0–2.5 min 2% B, 2.5–3 min 10% B, 3–7.5 min 15% B, 7.5–8.5 min 35% B, 8.5–9.5 min 98% B, and 9.5–10.0 min 2% B at a flow rate of 0.5 mL/min was used. A multiple reaction monitoring (MRM) approach was taken for the mass spectrometric determination of the polyphenols using argon as collision gas. The parameters for MRM transitions were obtained using the Waters integrated Intellistart^TM software (Waters Corp., Milford, MA, USA) (Table 1).
The UHPLC-MS/MS data were acquired using electrospray ionisation in negative ion mode (ESI–) with the following ionisation conditions: capillary voltage 3 kV, cone voltage 42 V, extractor voltage 3 V, source temperature 150 °C, desolvation gas flow 1200 L/h, and desolvation temperature 350 °C.