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摘 要：目的：建立用高效液相色谱法同时测定鸟嘌呤、胞嘧啶和胸腺嘧啶的方法，并用此方法对武昌鱼、草鱼、鲢鱼和鲫鱼中的含量进行测定。方法：采用HPLC对标样和样品进行测定；用盐酸溶液作溶剂，用超声提取法对鱼鳞进行提取。结果：鸟嘌呤、胞嘧啶和胸腺嘧啶分别在2.5×10-4 μg~5.2×10-3 μg(R=0.9999)、3.52×10-4 μg~4.41×10-3 μg(R=0.9999)和4.8×10-4 μg~4.8×10-3 μg(R=0.9999)线性范围内呈现良好的线性关系，相对标准偏差RSD分别为0.15 %、1.42 %、0.01 %；在超声条件下，0.5 M的盐酸对鱼鳞中的嘌呤类化合物提取率最高。结论：建立了用HPLC同时测定淡水鱼鱼鳞中嘌呤和嘧啶类化合物含量的方法。本方法准确度和精密度高，重现性良好；淡水鱼鱼鳞中均含有鸟嘌呤和胞嘧啶，并且鸟嘌呤含量较高。淡水鱼鱼鳞是提取鸟嘌呤的较佳原材料。 60903
毕业论文关键词：淡水鱼鱼鳞，鸟嘌呤，胞嘧啶，胸腺嘧啶，HPLC,同时测定
Abstract:Objective: This paper has established a method for simultaneous determination of guanine,cytosine,and thymine compound by high performance liquid chromatography (HPLC) and they were determined in Wuchang fish, Grass carp, Silver carp and Carassius fish scales. Methods: Standard samples and samples were determined by HPLC; The fish scales wered extracted by using ultrasonic assisted hydrochloric acid. Results: Guanine, cytosine and thymine showed good linearity in the ranges of 2.5×10-4~5.2×10-3μg(R=0.9999), 3.52×10-4 ~4.41×10-3μg (R
=0.9999),and4.8×10-4μg~4.8×10-3μg(R=0.9999) respectively, the relative standard deviation RSD were 0.15 %, 1.42 %, 0.01 %; The extracted ratio of guanine in fish scales was highest by 0.5 M hydrochloric acid. Conclusions: Purine and pyrimidine substances in freshwaterfish scales were examined simultaneously by the established method. The method was proved to be accurate and precise and showed good repeatability. All kinds of freshwater fish scales in the study contained both guanine and cytosine and higher content of guanine. The freshwater fish scales are the better raw materials for extracting guanine.
Keywords: freshwater fish scales, guanine, cytosine, thymine, HPLC, examine simultaneously
1 引言 4
2 实验部分 4
2.1 材料、仪器与试剂 4
2.2 高效液相色谱条件 5
2.3 标准曲线的绘制 5
2.4 精密度的考察 6
2.5 检出限和线性范围的测定 6
2.6 鱼鳞中嘧啶和嘌呤类化合物的提取及含量测定 7
2.7 加标回收率试验 7
2.8 重复性考察 8
3 结果与讨论 8
3.1 紫外波长的选择结果 8
3.2 标准曲线的绘制结果 9
3.3 精密度考察结果 11
3.4 检出限和线性范围的测定结果 11
3.5 鱼鳞中嘧啶和嘌呤类化合物提取方法的确定 13
3.6 加标回收率试验结果 16
3.7 重现性考察结果 16
结 论