Chirasil dex cb column

A 1 mL vial, under inert atmosphere, was filled with NaOH (1.0 mg, 25 μmol), acetophenone (12 μL, 100 μmol), and a solution of ruthenium complex 9 (1.3 mg, 1 μmol) in ⁱPrOH (0.125 mL, 1.63 mmol, around 16 equiv./ketone). The reaction mixture was heated for 1 h. The solution was diluted with 0.2 mL of CH₂Cl₂ and passed through a Millipore filter. An aliquot was analyzed by GC with a Chirasil-DEX CB column (25 m × 0.25 mm) (Agilent Technologies, Santa Clara, CA, USA) to determinate the enantiomeric excess. The remaining solution was concentrated under vacuum and the resulting crude solution was analyzed by ¹H NMR spectroscopy to determinate the conversion.

¹H NMR spectra were acquired on a Bruker-400 spectrometer in CDCl₃, and all signals were reported in parts per million (ppm) downfield relative to tetramethylsilane. Optical rotations were measured with a Perkin Elmer 341 polarimeter. Gas chromatographic analyses were performed on a Thermo Scientific TRACE 1300 gas chromatograph equipped with a CHIRASIL-DEX CB column (Agilent Technologies, USA), and using a flame ionization detector, nitrogen was used as the carrier gas at 5 mL/min, the split ratio was 1:20 (v/v), and the column temperature was programmed as being kept at 50 °C for 1 min and then upgraded to 150 °C at the rate of 3 °C/min. The protein purities were analyzed using SDS-PAGE using 4–12% polyacrylamide gradient gel (SurePAGE, Genscript, Nanjing, China) running in 50 mM MOPS, 50 mM Tris base, 0.1% SDS, and 1 mM EDTA, at pH 7.7, and stained with Coomassie Blue. Protein concentrations were measured using the Bradford assay (Coomassie Brilliant Blue G-250) and bovine serum albumin as the protein standard.