HPLC法检测哈巴苷及其黑曲霉转化产物

Determination on Harpagide and Its Transformed Products of Aspergillus niger by HPLC Method

[目的]研究黑曲霉转化液中哈巴苷及其3个转化产物的同时检测方法。[方法]通过用HPLC-SPE法快速测定黑曲霉转化液中哈巴苷及其3个转化产物的含量考察了该法的应用条件。[结果]哈巴苷、哈巴苷代谢素Ⅰ、哈巴苷代谢素Ⅱ和桃叶珊瑚甙B的线性回归方程分别为y=52.32x-89.76、y=32.52x-45.26、y=14.73x-25.64和y=41.25x-33.71,最低检测浓度分别为0.020、0.010、0.015和0.015μg/ml。0.010、0.100和2.500μg/ml哈巴苷的回收率分别为103.0%、99.7%和99.9%,0.010、0.500和2.500μg/ml的哈巴苷代谢素Ⅰ的回收率分别为102.1%、99.8%和99.9%,0.015、0.500和2.500μg/ml的哈巴苷代谢素Ⅱ的回收率分别为99.3%、99.6%和99.6%,0.015、0.500和2.500μg/ml的桃叶珊瑚甙B的回收率分别为100.6%、100.7%和99.9%。该法检测哈巴苷、哈巴苷代谢素Ⅰ、哈巴苷代谢素Ⅱ和桃叶珊瑚甙B的相对标准偏差在1.3%～4.7%。[结论]该方法能够快速、高效地检测黑曲霉转化液中哈巴苷及其转化产物的含量。

[Objective]The purpose was to study the simultaneous determination method of harpagide and its 3 transformed products in the transformation liquid of Aspergillus rage.[Method]The application conditions of HPLC-SPE method were investigated through determining the contents of harpagide and its 3 transformed products in the transformation liquid of A.niger rapidly by this method.[Result]The linear regression equations of harpagide,harpagidemetabolineⅠ,harpagidemetabolineⅡand aucubin B were y = 52.32x - 89.76,y = 32.52x - 45.26,y = 14.73x-25.64 and y = 41.25x -33.71 resp.Their minimum detectable concn.were 0.020,0.010,0.015 and 0.015μg/ml resp.The recoveries of 0.010,0.100 and 2.500μg/ml harpagide were 103.0%,99.7%and 99.9%resp.,that of 0.010,0.500 and 2.500μg/ml harpagidemetabolineⅠwere 102.1%,99.8%and 99.9%resp.,that of 0.015,0.500 and 2.500μg/ml harpagidemetabolineⅡwere 99.3%,99.6%and 99.6%resp.and that of 0.015,0.500 and 2.500μg/ml aucubin B were 100.6%,100.7%and 99.9%resp. The RSDs of determination on harpagide,harpagidemetabolineⅠ,harpagidemetabolineⅡand aucubin B by this method were in 1.3%～4.7%.[Conclusion]The contents of harpagide and its transformed products in the transformation liquid of A.niger could be determined rapidly and efficiently by this method.