动物营养学实验课程中色谱技术研究性实验设计

Researching Experiment Design of Chromatography Technology in Animal Nutrition Experimental Course

  • 摘要: 色谱技术是动物科学专业学生学习《动物营养学实验》课程时,开展研究性实验的重要环节。学生团队在教师的指导下,结合科研项目和生产实际,独立设计实验方案,成功建立了一种利用高效液相色谱法测定饲料调味剂中GMP和IMP含量的方法。样品经前处理后,采用Athena C18色谱柱,以甲醇和0.05 mol/L磷酸二氢钾溶液(2:98,v/v)作为流动相,流速设定为0.6 mL/min,进行等度洗脱。色谱柱的温度控制在25 ℃,检测波长设定为250 nm,进样量为10 μL。实验结果表明,GMP和IMP的浓度在0.05 mg/mL至2.0 mg/mL范围内与峰面积呈现良好的线性关系(R2=0.9999),其中IMP的检测限为0.4 μg/mL,GMP的检测限为0.8 μg/mL。此外,该方法的回收率高达98%,相对标准偏差分别为0.81%和0.75%。该实验项目不仅操作简便,分析时间合理,而且具有高精密度和回收率高,有助于激发学生的学习兴趣和探索精神,同时也有利于培养学生的创新能力和科学素养。

     

    Abstract: Chromatography is an important part of the research experiments by students in the major of Animal Science when they study the course Animal Nutrition Experiment. Under the guidance of the teachers, the student team independently designed the experimental scheme by combining the scientific research project and the actual production, and successfully established a method for the determination of GMP and IMP in feed flavoring agent using high-performance liquid chromatography. After the sample was pre-treated, the isocratic elution was carried out on an Athena C18 column with methanol and 0.05 mol/L potassium dihydrogen phosphate solution (2:98, v/v) as the mobile phases, and the flow rate was set at 0.6 mL/min. The temperature of the chromatographic column was controlled at 25 ℃, the detection wavelength was set at 250 nm, and the injection volume was 10 μL. The experimental results showed that the concentrations of GMP and IMP showed a good linear relationship with the peak areas in the range of 0.05 mg/mL to 2.0 mg/mL (R2=0.9999), with the detection limits of IMP being 0.4 μg/mL, and that of GMP being 0.8 μg/mL. In addition, the recoveries of the method were as high as 98% with the relative standard deviations of 0.81% and 0.75%, respectively. The experimental program is not only easy to operate and the analysis time is reasonable, but also has high precision and high recovery, which can help to stimulate the students’ interest in learning and the spirit of exploration, as well as cultivate their innovative ability and scientific literacy.

     

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