Deae sephadex a 25 40 mg column pyridine acetate form

The glucosinolates were extracted and analyzed as described by Sun et al. [17 (link)]. Briefly, the freeze-dried samples of four stages were boiled for 20 min in 1 mL of distilled water; the supernatant was collected after centrifugation (7000× g, 5 min), and the extraction step repeated once; the supernatants were mixed before being applied onto an analytical column using a DEAE-Sephadex A-25 (40 mg) column (pyridine acetate form) (GE Healthcare, Piscataway, NJ, USA). The glucosinolates were converted into their desulpho analogs after treatment with 200 μL of 0.1% aryl sulphatase for 16 h at room temperature. The resulting desulpho glucosinolates were then eluted with two washes (0.5 mL) of water, and the eluates were mixed and analyzed using a Waters HPLC instrument equipped with a model 2996 PDA absorbance detector (Waters, Milford, MA, USA). The samples were separated on a Waters Symmetry C18 column (250 × 4.6 mm i.d.; 5 μm) and data obtained with the absorbance at 226 nm. Ortho-nitrophenyl-β-d-galactopyranoside was used as an internal standard, and the normalization of peak-areas was used to calculate the glucosinolates.