Supplemental file 1
Preparative HPLC method for purification of the anthocyanins from Del/Ros1
tomato transgenic line
A W600 pump system (Waters) with a preparative HPLC column (X-Bridge MS C18,
5µm, 10x150mm, Waters), maintained at 24°C was used for separation. The mobile
phase was composed of 87% water, 3% ACN and 10% acetic acid (solvent A) as
well as 40% water, 50% ACN and 10% acetic acid (solvent B) at a flow rate of 2
ml/min. For elution the following gradient was used: initial 5% B; 80 min, 40% B; 8590 min, 100% B; 98 min, 5% B. Absorbance spectra were recorded with a
photodiode array detector (996, Waters) every 1 s, between 210 and 600 nm, with a
bandwidth of 1.2 nm, and chromatograms were acquired at 280 and 535 nm. Data
were analysed using Waters Empower software. Separated anthocyanins were
collected and the fractions were subjected to analytical HPLC analysis, as described
above, for confirmation.
HPLC methods for profiling of hydrolysate
Hydrolysates were analysed by HPLC (Alliance, Waters, Eschborn, Germany)
combined with a photodiode array detector (996, Waters). Separations were carried
out by reverse-phase chromatography on an Aqua C18, 5 µm, 4.6x250 mm column
(Phenomenex, Aschaffenburg, Germany), maintained at 25°C. The mobile phase
was composed of 87% water, 3% ACN and 10% acetic acid (solvent A) as well as
40% water, 50% ACN and 10% acetic acid (solvent B) at a flow rate of 1 ml/min. The
gradient was as follows: initial 6% B; 20 min, 20% B; 35 min, 40% B; 40 min, 60% B;
45 min, 90% B; 60 min, 6% B.