高效液相色谱法测定地顶孢霉培养物中的腺苷

建立测定虫草源饲料添加剂地顶孢霉培养物中腺苷含量的高效液相色谱方法。样品经过研碎、超声处理,离心、过滤后上机测定。腺苷的最佳提取条件:以超纯水为浸提液,用超声波浸提,浸提温度为40℃,浸提时间为55 min。使用Waters Spherisorb ODS2柱(150 mm×3.9 mm,5μm),以甲醇-0.01 mol/L磷酸二氢钾混合液(10∶90)为流动相,流量为1.0 mL/min,进样体积为20μL,检测波长为254 nm。腺苷的质量浓度在0.5~100μg/mL范围内与色谱峰面积成良好的线性关系,相关系数为0.9990。样品测定结果的相对标准偏差为1.65%(n=6),3水平加标的平均回收率为98.19%。该方法简便、快捷,可准确测定虫草饲料添加剂地顶孢霉培养物中腺苷的含量。

Determination of adenosine in the culture of acremonium terricola by high performance liquid chromatography

A HPLC method was established to determine adenosine in the culture of Acremonium terricola.After crushing,ultrasonic treatment,centrifugation and filtration,samples were detected by HPLC.The best extraction conditions:ultra-pure water as the immersion liquid,extrstion by ultrasonic crusher power,immersion temperature of 40℃,immersion time of 55 min.Waters Spherisorb ODS2 column(150 mm×3.9 mm,5μm)was used and methanol-0.01 mol/L potassium dihydrogen phosphate solution(10∶90)was used as the mobile phase,the flow rate was 1.0 mL/min,the sampling amount was 20μL and the detection wavelength was 254 nm.The mass concentration of adenosine was linear with the chromatograph peak area in the range of 0.5-100μg/mL with the correlation coefficient of 0.9990.The relative standard deviation of the detection results was 1.65%(n=6),and the average recovery of three level standard addition was 98.19%.The established method is simple,rapid and accurate,which can be used for the quality control of adenosine in the culture of Acremonium terricola.