对照品替代法用于缓解视疲劳片叶黄素含量测定的研究

目的：建立对照品替代法测定缓解视疲劳片中叶黄素的含量。方法：采用HPLC法，在不同条件下测定替代对照品苏丹Ⅰ对叶黄素对照品的校正因子，利用相对校正因子f和替代对照品进行叶黄素的含量测定。色谱条件：采用Shim-pack CLC-ODS色谱柱（6.0 mm×150 mm，5 μm），以甲醇-水（95：5）为流动相，流速1.0 mL·min^-1，检测波长473 nm，柱温30 ℃。结果：叶黄素和苏丹Ⅰ浓度均在0.001~0.05 mg·mL^-1范围内线性关系良好，相关系数分别为0.9999和0.9996；相对校正因子f=0.46；波长、流动相及不同色谱柱对f值没有显著影响；用对照品外标法和替代对照品法2种方法测定的结果无显著性差异，测得平均回收率均在95.6%~103.6%，RSD≤4.0%。结论：采用苏丹Ⅰ代替叶黄素作为对照品，用HPLC法测定叶黄素含量，进行产品质量控制，是经济、可行、实用的。

Objective: To establish and validate a method with a substitute for the reference substance for quantitative determination of lutein in visual fatigue relieving tablets. Methods: HPLC method was adopted.Sudan Ⅰ was selected as the substitute for reference substance of lutein and the correction factor of Sudan Ⅰ vs.the reference substance of lutein was determined under different conditions.The content of lutein was determined by the relative correction factor f and the substitute reference substance.The separation was performed on a Shim-pack CLC-ODS column(6.0 mm×150 mm,5 μm)at 30 ℃ using methanol-water(95:5)as the mobile phase at a flow rate of 1.0 mL·min^-1,and the detection wavelength was set at 473 nm. Results: Good linear correlation of lutein and Sudan Ⅰ was observed within the range of 0.001-0.05 mg·mL^-1;the correlation coefficients were 0.999,and 0.9996,respectively.The relative correction factor (f)was 0.46.Different wavelengths,mobile phases and columns had no significant effects on f values.There were no significant differences between the two quantitative methods by external standardization with reference substance and by reference substance substitution.The average recoveries were 95.6%-103.6% with RSD≤4.0%. Conclusion: The results indicated that the HPLC method by substitute reference substance was feasible,reliable and economical for quantitative determination in product quality control.