Peonidin chloride

Promega’s ‘Ultra-pure’ UDP-galactose (UDP-Gal) and UDP-glucose (UDP-Glc) were used as sugar donors in UGT assays (Madison, WI, USA). Cyanidin chloride, peonidin chloride, malvidin chloride, quercetin, kaempferol, isorhamnetin, naringenin, catechin, epicatechin, caffeic acid, chlorogenic acid, quinic acid, coumaric acid and ferulic acid standards were purchased from Sigma Aldrich (St. Louis, MO, USA). All standards were of HPLC grade and diluted to 100 mM in DMSO and stored at –20 °C until use. Acetonitrile, methanol, and formic acid (LC-MS grade) were purchased from Fisher Scientific (Waltham, MA, USA).

The anthocyanin extracts from the samples were analyzed for anthocyanins profile using HPLC (Agilent 1260 LC System, 5301 Stevens Creek Blvd Santa Clara, CA 95051, United States). A Gemini C₁₈ ODS column was used for HPLC analysis, and the detector was set at 520 nm (36 (link)). An injection volume of 20 μl with a 1 ml/min flow rate and the oven temperature of 35°C were used. The eluents were mobile phase A (water/acetonitrile/formic acid, 87/3/10 v/v/v, FINAR Ltd./Sisco Research Laboratories Pvt. Ltd., Mumbai, India) and mobile phase B (100% HPLC grade Acetonitrile, Sisco Research Laboratories Pvt. Ltd.). The chromatographic conditions were: 3% B in A at the time of injection at 45 min; 25% B in A at 46 min; 30% B in A at 47 min; 3% B in A (initial conditions). The anthocyanin standards Cyanidin-3-O-glucoside, Cyanidin-3-O-galactoside, Cyanidin chloride, Peonidin chloride, Pelargonidin chloride, and Malvidin chloride were obtained from Sigma Aldrich, United Kingdom, to identify and quantify anthocyanin fractions.