高效液相色谱–光电二极管阵列法测定虾青素的含量
Determination of Astaxanthin by High Performance Liquid Chromatography with Photodiode Array Detector
Abstract
建立虾青素含量测定的高效液相色谱–光电二极管阵列法。采用PurOSPHEr STAr rP 18(4.6 MMx250MM,5μM)色谱柱,以甲醇–水(体积比为95∶5)为流动相,流速1.0 Ml/MIn,检测波长为482 nM,柱温为30℃,进样量为20μl。在所选定的液相色谱条件下,虾青素主峰与其它杂质峰分离良好,虾青素在0.2~16μg/Ml范围内线性良好,线性相关系数r=0.999 9,检出限为0.01μg/Ml,测定结果的相对标准偏差为0.42%(n=6),平均回收率为100.4%。该法分析快速准确、灵敏度高、重现性好。 A method using high-performance liquid chromatography(HPLC) coupled with photodiode array detector(PDA) was established for determination of astaxanthin.The separation was performed on Purospher STAR RP 18 column(4.6 mm×250 mm,5μm) with the mobile phase of methanol–water(Volume ratio was 95∶5).The detector was photodiode array detector and the variable wavelength detector(VWD) was set at 482 nm.The column temperature was 30℃ and the injection volume was 20 μL.The calibration curve was linear in the range of 0.2–16 μg/mL,and the correlation coefficient was 0.999 9.The relative standard deviation of determination results was 0.42%(n=6).The average recovery was 100.4%, and the detection limit was 0.01 μg/mL.This method is characteristic of rapidity,accuracy,high sensitivity and good reproducibility.