高效液相色谱(HPLC)法测定紫花苜蓿叶片中叶黄素循环组分的含量

Determination of the Content of Xanthophyll Cycle Components in Leaves of Medicago sativa L. by HPLC

【目的】建立适合紫花苜蓿(Medicago sativa L.)叶片叶黄素循环组分:紫黄质(V)、环氧玉米黄质(A)和玉米黄质(Z)含量测定的方法。【方法】采用高效液相色谱(HPLC)法对紫花苜蓿叶片叶黄素循环的3个组分进行了测定,并对试验方法进行了验证。【结果】V、A和Z三个组分的标准品在3~100 μg/mL范围内均与峰面积呈良好的线性关系,相关系数分别为r=0.999 49、r=0.999 61和r=0.999 60,精密度试验(n=5)的RSD分别为0.96%、0.39%和0.87%,稳定性试验(n=7)的RSD值分别为3.33%、1.42%和1.86%,重现性试验(n=5)的RSD分别为0.48%、3.98%和2.59%,平均回收率(n=3)分别为99.55%、96.04%和94.08%,RSD分别为0.96%、1.38%和0.51%。【结论】HPLC法适于紫花苜蓿叶片叶黄素循环组分V、A和Z含量测定,该方法精密度高、重复性和稳定性好,回收率高。

[Purpose] To provide an effective method which can accurately determine xanthophyll cycle components, violaxanthin (V), antheraxanthin (A) and zeaxanthin (Z) in alfalfa (Medicago sativa L.) leaves.[Methods] Under high performance liquid chromatography (HPLC) method, the content of xanthophyll cycle components in alfalfa leaves was determined and the experimental method was validated.[Result] The content of xanthophyll cycle components was around 3-100 μg/mL and it had good linear relationship with the peak area, the correlation coefficients of V, A and Z were r=0.999 49, r=0.999 61 and r=0.999 60, respectively. The RSD of precision (n=5), stability (n=7) and reproducibility (n=3) of V, A and Z were 0.96%, 3.33% and 0.48%;0.39%, 1.42% and 3.98%;0.87%, 1.86% and 2.59%, respectively.[Conclusion] This method is efficient with high precision, reproducibility, stability and high recovery which makes it suitable for determine xanthophyll cycle components in alfalfa leaves.