Rapid determination of nine polymethoxyfl avones in sweet oranges juice using SPE-HPLC-DAD/FLD method

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  • 1 (Citrus Research Institute, Southwest University, Chongqing 400712, China) 2(Chongqing Collaborative Innovation Center for Functional Food, Chongqing University of Education, Chongqing 400067, China) 3(College of Food Science, Southwest University, Chongqing 400715, China) 4(College of Plant Protection, Southwest University, Chongqing 400715, China) 5(College of Biological and Chemical Engineering, Chongqing University of Education, Chongqing 400067, China)

Online published: 2018-02-06

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Abstract

A method using solid phase extraction (SPE) for sample preparation and followed with high performance liquid chromatography coupled with diode array detector and fluorescence detector (HPLC-DAD/FLD) analysis was developed to determine 9 polymethoxyflavones (PMFs) compounds in orange juice. Orange juice was filtered and went through a C18 SPE cartridge; the cartridge was washed in series with ultra-pure water and 30% of acetonitrile; the adsorbed components were then eluted with ethyl acetate, dried under nitrogen current and re-dissolved in methanol [l1] [ql2] to produce the PMFs sample. Using 0.05% of phosphoric acid solution, methanol and 50% of tetrahydrofuran as the mobile phase[l3] [ql4] , the sample was separated under gradient elution condition by reverse phase HPLC. Meanwhile, sample PMFs were qualitatively and quantitatively determined using the ultraviolet (UV) and fluorescence (FL) spectra and the peak area obtained by DAD and FLD. All the nine PMFs separated in 10 min under the solvent gradient composed. Using 330 nm for UV quantification, 340 nm and 450/500 nm for FL excitation and emission, the external standard calibration showed good linearity (R2>0.999). The converted limit of quantitation for juice was [l5] 4.00-61.2 μg/L and 0.024-0.61 mg/L respectively for UV and FL signal; recovery study showed good accuracy for both UV (96.4%-104.2%) and FL (94.2%-102.1%) examination; furthermore, UV and FL determination showed well accordance with each other. This method possessed the advantages including simple preparation, short time-consuming and low cost, which enabled it applicable for the rapid determination of PMFs contained in orange juice. As an instance application, fresh squeezed juices from eleven juice-processing sweet orange cultivars were analyzed; the results showed that Yuzaocheng contained the highest amount of PMFs, exhibiting prospective potency for further exploitation.

Cite this article

LI Gui-jie et al . Rapid determination of nine polymethoxyfl avones in sweet oranges juice using SPE-HPLC-DAD/FLD method[J]. Food and Fermentation Industries, 2018 , 44(1) : 245 . DOI: 10.13995/j.cnki.11-1802/ts.014148

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