关键词: 高效液相色谱, 一测多评, 葱莲, 芦丁, 异槲皮苷, 烟花苷, 紫云英苷

Abstract:

Objective： To establish a method for simultaneously determining the contents of four chemical components （rutin， isoquercitrin， nicotiflorin and astragalin） in Zephyranthes candida flowers by high performance liquid chromatography combined with the quantitative analysis of multi-components by single marker（HPLC-QAMS）.Methods： The HPLC-QAMS method was adopted. The chromatographic column was Lubex Kromasil C₁₈ （250 mm × 4.6 mm， 5 µm）. The mobile phase was a gradient elution of acetonitrile-0.1% phosphoric acid aqueous solution. The detection wavelength was 350 nm. The flow rate was 1.0 mL·min^-1. The column temperature was 30 ℃. The injection volume was 10 µL. Using rutin as the internal reference substance， the relative correction factors of isoquercitrin， nicotiflorin and astragalin relative to rutin were calculated by using the slope correction， multi-point correction and single-point correction methods respectively. The chromatographic peaks were located by the relative retention time. The difference between the results of the QAMS and the external standard method was investigated by the relative error （RE）， and the accuracy and feasibility of the quantitative analysis of multi-components by single marker were evaluated.Results： The linear ranges were 0.279-5.580 µg for rutin， 0.013-0.260 µg for isoquercitrin， 0.208-4.160 µg for nicotiflorin， and 0.019-0.372 µg for astragalin（r ≥ 0.999 9）. The RSDs of the precision， stability and repeatability tests were all less than 2%（n = 6）. The range of the average sample addition recoveries was 97.12% - 98.47% （RSD < 2%， n = 9）. There was no significant difference between the results of the QAMS method using slope correction and multi-point correction and those of the external standard method， and the results of the QAMS method using single-point correction were completely consistent with those of the external standard method.Conclusion： The established content determination method is simple， rapid， accurate and reliable， and can be used to simultaneously determine the contents of the 4 active components in Zephyranthes candida flowers.

Key words:  , High performance liquid chrormatography , （HPLC）； , Quantitative analysis of multi-components by single marker , （QAMS）； , Zephyranthes candida； , Rutin； , Isoquercitrin； , Nicotiflorin； , Astragalin