A C18 column (Phenomonex, SphereClone 5μ ODS(2)) was equilibrated for 3 h with a mobile phase which consisted of 80 mL (0.02 M) TBA (tetrabutylammonium bromide) and 20 mL ACN (acetonitrile) with detection at 229 nm. The flow rate was set at 1.0 ml/min and separated according to programme for desulfoglucosinolates detailed in Table 3 .
In the case of glucoraphasatin in R. sativus leaves and glucotropaeolin in B. juncea minor alterations were made to avoid peaks co-eluting. The mobile phase programme for R. sativus leaves was 100% A for 5 min, followed by a 35 min linear gradient to 66% B followed by a 5 min linear gradient to 100% B followed by a 5 min linear gradient to 100% A. For B. juncea leaves, an isocratic 85:15, TBA (0.02 M):acetonitrile mobile phase for 70 min was used.
Solution A: 100% TBA (0.02 M)
Solution B: 70:30, TBA (0.02 M):acetonitrile
In the case of glucoraphasatin in R. sativus leaves and glucotropaeolin in B. juncea minor alterations were made to avoid peaks co-eluting. The mobile phase programme for R. sativus leaves was 100% A for 5 min, followed by a 35 min linear gradient to 66% B followed by a 5 min linear gradient to 100% B followed by a 5 min linear gradient to 100% A. For B. juncea leaves, an isocratic 85:15, TBA (0.02 M):acetonitrile mobile phase for 70 min was used.
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