麦角菌发酵物中α麦角隐亭的RP-HPLC测定方法

RP-HPLC assay for the determination of α-ergocryptine in the ergot fermentation products

目的 :建立麦角菌发酵产物α麦角隐亭的RP HPLC定量测定方法 ,对在一定条件下α麦角隐亭发酵生产的峰值进行初步探索。方法 :色谱柱 (2 5 0mm× 4.6mm ,填料 :YWG C18,10 μm) ;流动相为甲醇 磷酸盐缓冲液 (pH7.0 ) 乙酸乙酯 (75∶2 5∶1) ;流速为 1.0ml·min-1;检测波长为 312nm。建立回归方程 ,进行方法精密度测定和加样回收率试验。结果 :回归方程A =190 5 8.5 4M +135 2 .6 6 ,r =0 .9997,在 0 .6 375～ 6 .4375 μg范围内线性关系良好 ;日内和日间相对标准偏差 (RSD)分别为1.15 %和 0 .44 % ;标准品浓度为 12 2 ,2 44和 488μg·ml-1时加样回收率分别为 10 8.48% ,10 5 .15 % ,10 9.5 3%。用该方法测得α麦角隐亭发酵生产的峰值一般在发酵的第 15～ 17d。结论 :该方法在定量测定麦角菌发酵产物α麦角隐亭方面灵敏、准确。

OBJECTIVE:To develop a reversed phase HPLC method for the quantitative determination of the ergot fermentation product α ergocryptine and to determine the peak value of α ergocryptine yielded during the fermentation.METHODS:A YWG C 18 column (250 mm×4.6 mm ID,particle size,10 μm) was used;the mobile phase was MeOH phosphate buffer (pH7.0) EtOAc (75∶25∶1) with a flow rate of 1.0 ml·min -1 and detected at 312 nm.A regression equation was set up,the precision of the method and the recovery rates were measured.RESULTS:Regression equation was A=19058.54M+1352.66,r=0.9997,over the range of 0.6375 ～6.4375 μg;the RSD for the intra day and inter day was 1.15%,0.44%,respectively;the mean recoveries were 108.48%,105.15% and 109.53% versus the added authentic sample at the concentrations of 122,244 and 488 μg·ml -1 ,respectively.The peak value appeared between day 15 to day 17 under the given condition.CONCLUSION:The method is sensitive and reproducible for the quantitative determination of the ergot fermentation product α ergocryptine.