Abstract: In order to establish a method for the separation and preparation of β-cryptoxanthin from paprika, the crude β-cryptoxanthin was extracted by ultrasonic acetone extraction, saponificated and separated by silica gel column chromatography. Preparative high performance liquid chromatography was used to refine and purify β-cryptoxanthin. The structure of β-cryptoxanthin was identified by UV, ESI-MS, 1H-NMR and 13C-NMR. The purity of β-cryptoxanthin was determined by analytical HPLC. The results indicated that complete saponification of chili pigment extract was achieved when saponifying its dispersion in 45 mL of isopropanol (9 g/mL) with 6.5 mL of 20% KOH methanol solution for 24 h at room temperature. During the rough separation by silica gel column chromatography using petroleum ether and acetone as elution solvents, β-cryptoxanthin could be better enriched at a volume ratio of 10:1. The preparative high performance liquid chromatography required acetone and water at a ratio of 95:5 (V/V) and a flow rate of 5 mL/min. Under these optimized conditions, β-cryptoxanthin with a purity of 99.0% could be obtained. This developed method is simple, rapid, reproductive, and suitable for the preparation of β-cryptoxanthin with high purity.

Key words: paprika, β-cryptoxanthin, preparative HPLC