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目的:建立高效液相色谱法(HPLC)测定羊蹄大黄素含量的方法,并对酸性和碱性土壤环境中羊蹄各部位的大黄素含量进行比较。方法:采用超声提取法制备供试品溶液,运用高效液相色谱法测定羊蹄大黄素含量。Zorbax XDB-C18色谱柱;流动相为甲醇-1%冰醋酸(70∶30);检测波长:254nm;流速:1.5 mL.min-1;柱温:40℃;进样量:20μL。结果:大黄素进样在1μg.mL-1～500μg.mL-1浓度范围内与峰面积线性关系良好(r=0.9995),加样回收率为98.51%,RSD为2.04%(n=6)。酸性土壤条件下羊蹄根、根茎与叶中大黄素含量分别为1.5796mg.g-1、1.4634mg.g-1和0.1223mg.g-1。碱性土壤条件下羊蹄根、根茎与叶中大黄素含量分别为0.4918mg.g-1、0.3162mg.g-1和0.0806mg.g-1。结论:本法适用于羊蹄中大黄素含量的检测。酸性土壤条件下羊蹄大黄素含量明显高于碱性土壤条件,羊蹄根与根茎中大黄素含量高于叶中大黄素含量。
OBJECTIVE: To establish a HPLC method for the determination of emodin and to compare the emodin content of various parts of the milkvetch in acidic and alkaline soil environments. Method: Ultrasound extraction was used to prepare the test solution, and the contents of emodin in the samples were determined by high performance liquid chromatography. Zorbax XDB-C18 column; mobile phase was methanol-1% glacial acetic acid (70:30); detection wavelength was 254 nm; flow rate was 1.5 mL.min-1; column temperature was 40 ℃; injection volume was 20μL. Results: The emodin injection had a good linear relationship with the peak area (r = 0.9995) in the concentration range of 1μg.mL-1 ~ 500μg.mL-1, the recovery was 98.51% and the RSD was 2.04% (n = 6) . Emamectin, rhizome and leaf emodin contents in acid soil were 1.5796mg.g-1, 1.4634mg.g-1 and 0.1223mg.g-1, respectively. Under alkaline soil conditions, the contents of emodin in roots, rhizomes and leaves of F. graminearum were 0.4918mg.g-1, 0.3162mg.g-1 and 0.0806mg.g-1, respectively. Conclusion: This method is suitable for the detection of emodin in sheep’s hoof. The contents of emodin in acid soil were significantly higher than those in alkaline soil. The content of emodin in the roots and rhizomes was higher than that in the leaves.