Dertermination of Anthocyanidin Composition and Content from Rosa damascene tyigintipetala at Different Flowering Periods by HPLC

In this experiment, the HPLC method was used to determine 6 main anthocyanidins, including delphinidin pigment, cyanidin pigment,  pelargonidi pigment,  peonidin pigment, malvidin pigment and petunidin pigment, from Rosa damascene tyigintipetala at 4 different flowering periods (i.e., unfold-petal stage, initial-flowering stage, full-flowering stage and flowering-wilting stage). The anthocyanidins in the sample were extracted by ultrasonication for 30 min with the extracting solvent (ethanol : water : hydrochloric acid = 2 : 1 : 1), and hydrolyzed for 1 h in a boiling water bath. The HPLC was carried out by using an Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm), the flow phase was 1% formic acid aqueous solution and 1% formic acid acetonitrile solution, the flow rate was 0.8 mL·min⁻¹, the detection wavelength was 530 nm, and the column temperature was 35 ℃. The results showed that cyanidin and pelargonidin pigments were detected at 4 flowering periods from Rosa damascene tyigintipetala. Variance analysis showed that the content difference between these two anthocyanidins at different flowering stages was extremely significant (P<0.01). The content of these two anthocyanidins decreased with the flowering process and petal fading, and the content of cyanidin pigment was absolutely dominant at different flowering stages. The established extraction and detection method of anthocyanidins from Rosa damascene tyigintipetala in this study is simple, rapid, stable and reproducible, which can be further used as an effective method for the determination of 6 anthocyanidins, such as delphinidin pigment, cyanidin pigment,  pelargonidi pigment,  peonidin pigment, malvidin pigment and petunidin pigment from Rosa damascene tyigintipetala and other plant-derived materials.