Reversed‐phase and hydrophilic interaction modes of liquid chromatography determination of tulip petal anthocyanins

Abstract Tulip flower petals, often regarded as waste plant material, represent a rich and promising source of anthocyanins for industrial production. In this study, reverse‐phase (RP) chromatography and hydrophilic interaction liquid chromatography (HILIC) with a diol phase were evaluated for the separation of anthocyanins in tulip petals. It was found that reverse‐phase chromatography required gradient elution, whereas HILIC achieved effective separation using a simple isocratic elution. The diol phase in HILIC demonstrated the highest efficiency for anthocyanin separation using mobile phases acidified with either phosphoric or formic acid. In terms of anthocyanin composition in tulip petals, pelargonidin, cyanidin, and delphinidin were identified as the primary anthocyanidin backbones across eight tulip cultivars. The predominant glycosides were 3‐rutinosides and/or acetylated 3‐rutinosides, while 3‐glucosides were present at nearly trace levels. A specific solute wavelength was proposed for area determination in the internal normalization method, enabling relative quantification of anthocyanin content. The total anthocyanin accumulation in the petals was measured using the differential spectrophotometric method. Furthermore, solute retention was compared and analyzed against two types of descriptors—miLogP and total surface polar areas—for both elution types.