样品预处理方法对虫草头孢菌粉中腺苷含量测定的影响

Investigalion of the Effects of Sample Preparation on Delermination of the Content of Adenosine in Fermented Cordyceps Powder

为了建立一种利用RP-HPLC法精确测定虫草头孢菌粉中腺苷含量的方法,实验在现有色谱条件的基础上优化了流动相条件,通过研究提取溶剂、时间、温度对腺苷提取效果的影响,比较超声波提取和加热回流提取的效果,以此探索虫草菌粉中腺苷测定时样品预处理的方法.结果表明,色谱条件为Diamonsil^TM C18色谱柱（250×4.6 mm,5μm）、流动相乙腈-水（9∶91）、紫外检测波长260 nm时,能够将虫草菌粉中的腺苷和其他成分分开;腺苷质量分数在0.097-0.582 mg.mL-1范围内与峰面积呈良好线性关系;平均加样回收率为101.1%;RSD为2.20%.样品预处理方法以在室温下用体积分数50%的甲醇超声波提取60 min为最好,该方法所需时间短,操作简单.

To establish a method for determining the content of adenosine in fermented Cordyceps powder simply and accurately with RP- HPLC, in this experiment, based on the pre-established chromatogram condition, mobile phase condition was optimized, and the sample preparation of fermented Cordyceps powder was investigated by testing the influence of solvent, time, temperature on the extraction of adenosine and comparing the results of ultrasonic treatment and reflux. When the HPLC method was carried on DiamonsilTM C18 column （250 ×4.6 mm, 5 μm）, using acetonitrile -water（9：91 ） as a mobile phase with UV detector at 260 nm, adenosine was separated from other substances. The linearity of this method was good （r = 1） in the range 0.097 -0.582 mg· mL^-1 and average recovery was 101.1% with RSD = 2.20% （ n = 6）. The best sample preparation is to treat sample in 50% methanol solution for 60 minutes at room temperature with ultrasonic treatment. The method is time saving and easy to conduct.