Lc dad system

Identification was performed as previously described Ruiz et al. [12 (link)]. The same column, precolumn, and chromatographic separation used for anthocyanin analysis were used. The spectrophotometric detection was set at 320 nm, and MS detection was set using the BPC, both obtained in the LC-MS. The same spectrometric conditions used in Section 2.3.2 were used, with the exception of negative ionization ESI −3500 V. The quantitative analysis was carried out using a Shimadzu LC-DAD system by an external calibration curve prepared with 3-caffeoylquinic acid standard and measured at 326 nm.

Analytical HPLC was carried out using a Shimadzu LC-DAD system (Shimadzu, Kyoto, Japan; Degaser: DGU-20A3R prominence degassing Unit, Liquid Chromatograph: LC-20 AD prominence liquid chromatograph, SIL-20A HAT prominence autosampler, and SPA-M20A prominence diode-array detector) with a YMC-Pack Pro C18 column (S-5 µm, 12 nm, 150 × 10.0 mm). The preparative HPLC was performed according to Buckett et al. (2020) [43 (link)]. Regarding the biosynthesised compounds, each peak was collected using the analytical method and, after approximately 20 runs, this resulted in enough material for NMR.