Chromnav software v 2

One-dimensional and two-dimensional NMR spectra were acquired on an AVANCE III (400 MHz) (Bruker BioSpin, Rheinstetten, Germany), with chemical shifts expressed in ppm. The NMR spectra were referenced to residual solvent peaks (CD₃OD: ¹H NMR 3.30 ppm, ¹³C NMR 49.0 ppm). HR-ESI-MS spectra were acquired on a Thermo Fisher Scientific Q-Exactive HR-ESI-Orbitrap-MS (Waltham, MA, USA). Medium pressure liquid chromatography (MPLC) was conducted using an AI-580 system equipped with an ULTRA PAK ODS-SM-50D (50 µm, 50 × 300 mm, Yamazen Corporation, Osaka, Japan). Reversed-phase (RP)-HPLC separations were performed with a recycling system comprising a PU-2086 Plus Intelligent prep pump (Jasco, Tokyo, Japan), UV-2075 detector (Jasco, Tokyo, Japan), Capcell Pak UG120 C18 column (5 μm, 20 × 250 mm, Osaka Soda, Osaka, Japan), Capcell Pak UG120 C18 column (5 μm, 10 × 250 mm, Osaka Soda, Osaka, Japan), and HPLC-grade solvents. For analytical HPLC, a PU-4180 RHPLC pump (Jasco, Tokyo, Japan), MD-4017 photodiode array detector (Jasco, Tokyo, Japan), and AS-4050 HPLC autosampler (Jasco, Tokyo, Japan) were used. Data were analysed using ChromNAV software v.2 (Jasco, Tokyo, Japan).

Quantification of trimesic acid (H₃BTC) was performed using HPLC using a reversed phase Jasco LC-4000 series system, equipped with a PDA detector MD-4015 and a multisampler AS-4150 controlled by ChromNav software v.2 (Jasco Inc., Easton, MD, USA). A Purple ODS reverse-phase column (5 µm, 4.6 × 150 mm² Análisis Vínicos, Ciudad Real, Spain) was employed. The mobile phase consisted of a 50:50 solution (v/v) of buffer (0.04 M, pH = 2.5) and methanol (MeOH). The injection volume was set at 30 µL with a flow rate of 1 mL·min⁻¹ and the column temperature fixed at 25 °C. The standards used for the calibration curve consisted of trimesic acid solutions in MilliQ water solution with a concentration range from 9.65 to 0.01 µg·mL⁻¹ (correlation coefficient > 0.99). The chromatogram of the standard solution showed a retention time (rt) of 2.70 min (λ_max at 225 nm).