Eca 500 instrument

Monosaccharide analysis was performed as described previously (Urai et al., 2006 (link)). Briefly, GXM was hydrolyzed by 4 M TFA at 100°C for 3 h, and the monosaccharides obtained were labeled with aminobenzoic acid ethyl ester (ABEE) and analyzed by high performance liquid chromatography (HPLC) (1500 HPLC system, Waters, Milford, MA). All nuclear magnetic resonance (NMR) spectra were recorded at 500 MHz (¹H) with an ECA 500 instrument (JEOL Ltd. Tokyo, Japan). Chemical shifts are given in δ units with acetone (δ¹H 2.23) used as an external reference for samples measured in D₂O solutions. O-deacetylation and sonication of GXM were performed as described previously (Urai et al., 2006 (link)).

The starting materials, reagents, and solvents were purchased and used without further purification. We used Silica gel 70 F254 TLC plates (Wako) for analytical Thin-layer chromatography (TLC), while Silica gel 60 N (spherical, neutral, Kanto Chemical) were for column chromatography. For preparative flash chromatography, used were an automated system (Smart Flash EPCLC AI-580S, Yamazen Corp., Japan) equipped with universal columns of silica gel. Melting points and IR spectra were determined with a MP-500P (Yanaco) and a Nicolet 6700 spectrometer with anATR attachment, respectively. ¹H and ¹³C NMR spectra were determined on a JEOL ECA-500 instrument (500 MHz for ¹H and 126 MHz for ¹³C). Mass spectra were obtained with a high-resolution electro-spray ionization mass spectrometer, JMS-T100LC (JEOL). UV/visible absorption spectra were determined with a spectrophotometer, Cary 60 (Agilent Technologies) (scan speed 600 nm/min; data interval 1 nm). BL and chemiluminescence spectra were measured with a precision spectrophotometer, AB-1850 (ATTO) (data interval: 1 nm). BL intensities were monitored using luminometers, AB-2270 (ATTO) and GL-201A (Microtec Co.). BL imaging was performed with a multifunctional in vivo imaging system (IVIS Spectrum, PerkinElmer).

NMR spectra were recorded at 500 MHz (¹H) and 125 MHz (¹³C) using an ECA 500 instrument (JEOL Ltd., Tokyo, Japan) or at 600 MHz (¹H) and 150 MHz (¹³C) with an ECZ 600 instrument (JEOL Ltd.). Chemical shifts were administered in parts per million (ppm), with acetone (δ ¹H 2.23 ppm, δ ¹³C 31.1 ppm) used as an internal reference for samples measured in D₂O solutions. Signals were assigned based on the results of the heteronuclear single-quantum coherence (HMQC) and heteronuclear multiple-bond coherence (HMBC) experiments. ¹H NMR chemical shifts of the overlapping signals were obtained from the center of the cross peaks in the 2D spectra.