HPLC法测定哈蟆油中3β-羟基胆甾-5-烯-7-酮

Determination of 3β-hydroxy-cholesta-5-en-7-one in Oviductus Ranae by Using HPLC Assay

建立哈蟆油中3β-羟基胆甾-5-烯-7-酮的反相高效液相色谱含量测定方法。采用Aglient ZORBAX Bonus-RP（250mm×5mm5μm）色谱柱；检测波长：250nm；流动相：乙腈-水（87：13）；体积流量：1．0mL／min。3β-羟基胆甾-5-烯-7-酮在0．01004-0．11044μg与峰面积值呈良好线性关系（r=0．9998）。平均回收率为96．48％，RSD为1．66％。本方法简便、稳定、精密度高、重复性好，可用于测定哈蟆油中3β-羟基胆甾-5-烯-7-酮的含量。

To establish a HPLC method for the determination of 3 β-hydroxy-cholesta-5-en-7-one in Oviductus ranae. The experiment was carried out using Aglient ZORBAX Bonus-RP （250 mm× 5 mm 5 μ m）at the wavelength of 250 nm,and the mobile phase consisted of acetonitrile-water（87 ： 13 ） at a flow rate of 1.0mL/min. The calibration curve of 3 β -hydroxy-cholesta -5- en- 7-one in good linearity over the range of 0.010 4-0.110 44 μ g （r = 0.999 8 ）. The average recovery was 96.48 % and RSD was 1.66 %. The method is simple, stable, accurate and reproducible. It can be used for determining the content of 3β -hydroxy-cholesta-5-en-7-one in Oviductus ranae and can be used to control the qualities of Oviductus ranae.