Grapes at 8, 12, and 16 WAA of 5 cultivars were used to determine individual anthocyanins. Specific extraction steps have been described previously [20 (link)]. Three independent samples were extracted for each skin, and were stored at −40 °C until analysis. Before injection, the extracts were filtered through 0.45 µm filters (cellulose acetate and nitrocellulose, CAN).
Individual anthocyanins of extracts were analyzed using an Agilent 1100 series LC-MSD trap VL equipped with a diode array detector and a Kromasil C18 column (250 × 4.6 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA) utilizing a binary solvent gradient, where mobile phase A was 2% formic acid in water and B was 2% formic acid in acetonitrile. The detailed LC procedures and MS conditions have been described previously report [20 (link)].
The elution order and retention time of the anthocyanins were compared with those of the standard, and the weights of molecular ions and fragment ions were compared with the weights of standard and reference products, allowing different anthocyanins to be identified [25 (link),26 (link)]. Anthocyanins were quantified using malvidin-3-O glucoside as an external standard.
Individual anthocyanins of extracts were analyzed using an Agilent 1100 series LC-MSD trap VL equipped with a diode array detector and a Kromasil C18 column (250 × 4.6 mm, 5 μm) (Agilent Technologies, Santa Clara, CA, USA) utilizing a binary solvent gradient, where mobile phase A was 2% formic acid in water and B was 2% formic acid in acetonitrile. The detailed LC procedures and MS conditions have been described previously report [20 (link)].
The elution order and retention time of the anthocyanins were compared with those of the standard, and the weights of molecular ions and fragment ions were compared with the weights of standard and reference products, allowing different anthocyanins to be identified [25 (link),26 (link)]. Anthocyanins were quantified using malvidin-3-O glucoside as an external standard.
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