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The objective of this research is to present method development and validation for the simultaneous determination of lutein and zeaxanthin using ultra performance liquid chromatography (UPLC). Also, rapid quantification was performed on six green leafy vegetables (Allium tuberosum, Aster scaber, Hemerocallis fulva, Pimpinella brachycarpa, Sedum sarmentosum and Spinacia oleracea) that are commonly consumed in Korea. Separation and quantification were successfully achieved with a Waters Acquity BEH C18 (50×2.1 mm, 1.7 µm) column by 85% methanol within 5 min. Two compounds showed good linearity (r 2 >0.9968) in 1-150 µg/mL. Limit of detection (LOD) and quantification (LOQ) for lutein and zeaxanthin were 1.7 and 5.1 g/mL and 2.1 and 6.3 g/mL, respectively. The RSD for intra- and inter-day precision of each compound was less than 10.69%. The recovery of each compound was in the range of 91.75-105.13%. Aster scaber and Spinacia oleracea contained significantly higher amounts of lutein (4.06±0.24 and 3.97±0.10 mg/100 g of fresh weight), respectively.

Method Validation and Quantification of Lutein and Zeaxanthin from Green Leafy Vegetables using the UPLC System