2.6. Determination of glucosinolates
Finely ground seeds of each accession were analysed for their glucosinolate content according to the EU official ISO 9167-1 method (EEC, 1990) based on the high-performance liquid chromatography (HPLC) analysis of desulfo-GLs, with some minor modifications reported in Whatelet et al. (2004). The desulfo-GLs were analysed using an Agilent Model 1100HPLC system with an Inertsil ODS-3 column (250 × 3 mm, 5 μm), thermostated at 30 °C, with a diode array as detector (Lazzeri et al., 2011). Peak identification was performed comparing retention times and UV spectra of the samples with desulfo-GL standards available in our laboratory. The GL content was calculated using a quantified desulfo-sinigrin amount as internal standard, being sinigrin not present in Isatis genus, and its relative proportionality factor value ( Clarke, 2010). In particular, not being available in our laboratory the standard of the desulfo-glucoisatisin/epiglucoisatisin GLs present in the epimeric mixture, in order to quantify their content, a proportionality factor value of 0.2, similar to the ones used for indolic GLs and relative to the internal standard sinigrin was applied.
2.6. Determination of glucosinolatesFinely ground seeds of each accession were analysed for their glucosinolate content according to the EU official ISO 9167-1 method (EEC, 1990) based on the high-performance liquid chromatography (HPLC) analysis of desulfo-GLs, with some minor modifications reported in Whatelet et al. (2004). The desulfo-GLs were analysed using an Agilent Model 1100HPLC system with an Inertsil ODS-3 column (250 × 3 mm, 5 μm), thermostated at 30 °C, with a diode array as detector (Lazzeri et al., 2011). Peak identification was performed comparing retention times and UV spectra of the samples with desulfo-GL standards available in our laboratory. The GL content was calculated using a quantified desulfo-sinigrin amount as internal standard, being sinigrin not present in Isatis genus, and its relative proportionality factor value ( Clarke, 2010). In particular, not being available in our laboratory the standard of the desulfo-glucoisatisin/epiglucoisatisin GLs present in the epimeric mixture, in order to quantify their content, a proportionality factor value of 0.2, similar to the ones used for indolic GLs and relative to the internal standard sinigrin was applied.
การแปล กรุณารอสักครู่..