高效液相色谱法同时测定怀山药中4个山药素化合物

目的: 建立HPLC法同时测定怀山药不同部位(块茎、皮、茎叶、零余子)中4个山药素化合物(山药素Ⅰ、Ⅲ、Ⅳ、Ⅴ)的含量. 方法: 采用Aglient HC-C₁₈(4.6 mm×250 mm,5 μm)色谱柱,以甲醇(A)-水(B)为流动相,洗脱梯度(0~20 min,50%A;20~65 min,50%A→65%A),流速1.0 mL·min^-1,检测波长274 nm,柱温30℃,进样量20 μL. 结果: 4个山药素化合物在60 min内得到良好的分离,在给定的浓度范围内,4个山药素化合物呈现良好的线性(r≥0.999 9)、精密度(RSD≤0.93%)、稳定性(RSD≤0.62%)和重复性(RSD≤3.2%),回收率在91%~102%之间.其中怀山药的零余子和茎叶中均含有4个山药素化合物,山药皮中含有3个山药素化合物(山药素Ⅰ、Ⅲ、Ⅴ),块茎中含有2个山药素化合物(山药素Ⅰ、Ⅲ). 结论: 经方法学验证,本文建立的方法为怀山药质量标准研究、产品质量控制以及山药素类化合物的化感研究打下了基础.

Objective: To develop an HPLC method for simultaneous determination of four batatasins(Ⅰ,Ⅲ,Ⅳ,Ⅴ)in different parts of Dioscorea opposita Thunb.(tuber,peel,stem,bulbil). Methods: Separation was carried out on a reversed-phase Aglient HC-C₁₈ column(4.6 mm×250 mm,5 μm);the gradient mobile phase consisted of methanol(A)and water(B)with gradient elution(0-20 min,50%A;20-65 min,50%A→65%A)at the flow rate of 1.0 mL·min^-1;the column temperature was 30℃,the UV spectra detector was set at 274 nm and all the injection volume was 20 μL. Results: In the given concentration range,the batatasins showed good liner correlation(r≥0.999 9)with the RSD of precision lower than 0.93%,the RSD of stability lower than 0.62%,RSD of repeatability lower than 3.2%,and the recovery rate between 91%-102%.The results showed that the bubil and stem contained all the four batatasins,the peel contained three batatasins(batatasins Ⅰ,Ⅲ and Ⅴ)and the tuber contained two batatasins(batatasins Ⅰ and Ⅲ). Conclusion: Methodology verification showed that the experimental results laid a good foundation for the quality standard research of Chinese yam and the allelopathic effects of batatasin compounds.

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