Ga 55

For BM, an aliquot of 10 mL of 0.01 mol/L PBS pH 7.4 was added to 2 g of lyophilized BM and shaken at 200 rotation per minute (rpm) for 15 min. Subsequently, the volume was adjusted up to 16 mL with 0.01 mol/L PBS pH 7.4. For IPM, a volume of 20 mL of 0.01 mol/L PBS pH 7.4 was added to 4 g of IPM and 1.8 g of NaCl and shaken at 200 rpm for 15 min, and the volume was adjusted up to 25 mL with 0.01 mol/L PBS pH 7.4. After the BM and IPM samples were centrifuged at 1670× g for 20 min, the fat layer was removed, and the supernatant was filtered using two glass fiber filters (GA-200 followed by GA-55, Advantec, Tokyo, Japan). The AFM₁ extraction by IAC was carried out with 10 mL of 0.01 mol/L PBS pH 7.4, followed by 10 mL of reconstituted BM or IPM. The IAC was washed with 5 mL of PBS 0.01 mol/L PBS pH 7.4 and 5 mL of distilled water. The water inside the IAC was removed by adding air (30 mL). The elution was performed with 10 mL of CH₃OH:CH₃CN (1:9), and the eluate was dried in a rotary evaporator (Eyela, Tokyo, Japan). In this study, the rapid drying time was an important factor to maintain stability of AFM₁ for analysis.

The water absorption of each flour sample was determined using the technique developed by Matsuki et al. [23 (link)]. Seven holes (1.5 mm diameter) were created in the bottom of a column-shaped plastic container (4.7 cm × 9 cm × 1.5 mm), 12 mm apart. One piece of glass fiber filter paper (GA-55, 4.7 cm diameter, Advantec Tokyo Roshi, Tokyo, Japan) was placed inside the container. The flour sample (10 g) was weighted in the container. A weight (25 g) was placed onto the container to ensure that the sample did not float during the experiment. The container was placed in a tray filled with 1 cm-deep water to let water in through the holes at the bottom. A piece of filter paper (#2, 7.0 cm diameter, Advantec Tokyo Roshi, Tokyo, Japan) was placed under the container in the tray to avoid tight contact between the bottom of the container and the tray. The container was taken out of the water for weighing and was quickly placed back in the water. The amount of water in the flour sample was calculated from the moisture content of the flour.

The electrochemical test was performed in a non-aqueous electrolyte system as follows. Cathode electrodes were fabricated by mixing 90 wt% MM_NFPC/C with 10 wt% polytetrafluoroethylene (PTFE, Daikin Industries Ltd.) and then pressed into disks (loading amount: 30 mg/cm²). After drying the cathode pellet at 120 °C for 12 h under vacuum, the cathode properties were investigated in a 2032 coin-type cell with an organic electrolyte of 1 M NaPF₆/ethylene carbonate (EC): dimethyl carbonate (DMC) (1:1 in volume, Tomiyama Pure Chemical Industries). All coin cells were fabricated using a glass fiber separator (GA-55, Advantec), and Na metal (Sigma Aldrich) as the anode electrode in an Ar-filled glove box.
For the electrochemical test in 17 m NaClO₄ aq., cathode electrodes were assembled by mixing 90 wt% MM_NFPC/C with 10 wt% PTFE and then pressed into disks with the same loading of 30 mg/cm². NASICON-type NaTi₂(PO₄)₃ (denoted as NTP) as anode was synthesized by the solid-state method^{25 (link)}, and then mixed NTP with AB and PTFE in a weight ratio of 70:25:5. The cathode and anode pellets were assembled using Ti mesh as current collector (Thank Metal Co., Ltd.), 17 m NaClO₄ solution as the aqueous electrolyte (molality (m) = mole of solute/weight of solvent), and a silver-silver chloride electrode with saturated KCl (RE-6, BAS Inc.) as a reference electrode.

Biomass concentration was determined gravimetrically via filtration of cell suspension onto pre-weighed glass fiber filters (GA-55, Advantec) and subsequent drying and weighing. Optical density (OD) of cell suspensions was measured in 1-cm cuvettes at 750 nm with a DR3900 spectrophotometer (Hach).

Biomass concentration was determined by suspended solids dry weight measurements. A 10 mL aliquot of the microalgae suspension was filtered through a glass fiber filter paper (GA-55 Advantec, USA), and rinsed several times with UPW. The filter was then placed in oven at 80 °C for 24 h.