J w db 5ms capillary column

The composition of cellulose, hemicellulose, and lignin in non-pretreated, oxygen-gas-pretreated, and oxygen-radical-pretreated wheat straw was determined according to previously described methods [39 , 40 (link)]. Oxygen-gas- and oxygen-radical-pretreated wheat-straw samples were extracted with water to remove inhibitors for enzymatic reactions. Each treatment portion was washed separately at a weight ratio of 1:10 of pretreated wheat-straw samples to 25 °C Milli-Q water. The mixture was stirred at 100 rpm for 60 min, and the extract was filtered through a nylon membrane (pore size: 0.45 μm). The liquid fraction was lyophilized, trimethylsilylated, and analyzed using a GCMS-QP2010 (Shimadzu) equipped with a J & W DB-5MS capillary column (30 m × 0.25 mm internal diameter × 0.25 μm film thickness; Agilent Technologies, Santa Clara, California, USA) [41 (link)].

Sake meter value, sake acidity, and amino acid levels were measured as previously described⁶ . Organic acids were quantified using HPLC with a Shim-pack SCR-102H column (Shimadzu, Kyoto, Japan). Aroma components were quantified using headspace GC with a J & W DB-WAX capillary column (30 m × 0.32 mm internal diameter × 0.50 μm film thickness; Agilent Technologies, Santa Clara, California, USA). Ethanol concentration was measured using an ethanol analyser (RIKEN KEIKI, Tokyo, Japan). Sugars were determined by monitoring post-column derivative reducing sugars separated using a Prominence reducing-sugar HPLC analytical system (Shimadzu) equipped with a fluorescence detector. Sake fermented with MC87-46 and K901 was separated on a Shim-pack 4.0 × 250-mm ISA-07/S2504 column (Shimadzu) with a linear gradient of 0.1 M potassium borate buffer (pH 8.0) and 0.4 M potassium borate buffer (pH 9.0) for 140 min at a flow rate of 0.6 mL/min^{37 (link),38 (link)}. Sugars were also lyophilized, trimethylsilylated, and analysed using a GCMS-QP2010 (Shimadzu) equipped with a J & W DB-5MS capillary column (30 m × 0.25 mm internal diameter × 0.25 μm film thickness; Agilent Technologies)^{39 (link)}.

The GC-MS analysis was performed using an Agilent 7890B GC with the split-splitless inlet, coupled to a 7010 Triple Quadrupole (MS/MS) system from Agilent, equipped with EI ion source. The system was operated by MassHunter Software with MSD ChemStation. The separation was carried out on an Agilent J&W DB-5 ms capillary column (30 m × 0.25 mm i.d., 0.25 mm film thickness). Helium (purity 99.999%) was employed as carrier gas at a constant column flow of 1.0 mL/min. The GC oven temperature was programmed from 50 °C (held 1 min) to 270 °C at 10 °C/min (held 5 min). The temperature of the transfer line, the quadrupole and the ion source were set at 280, 150 and 230 °C respectively. The injector temperature was set at 250 °C. The injector was operating in the split-less mode and programmed to return to the split mode after 2 min from the beginning of a run. The MSD was operated in full scan acquisition mode. Mass spectra deconvolution of chromatographic signals and tentative identification of unknown bioactive compounds was carried out using the Agilent MassHunter Unknown Analysis tool and the NIST Mass Spectral Library (NIST MS Search 2.0).