【作者】 韩雯雯；

【导师】 刘盛辉； 胡宝祥； 莫卫民；

【作者基本信息】 浙江工业大学 ， 应用化学， 2007， 硕士

【摘要】 糖尿病是一种严重危害人体健康的常见慢性终身疾病，盐酸二甲双胍作为糖尿病的治疗药物，现已成为轻、中度Ⅱ型糖尿病患者特别是肥胖患者的首选治疗药。因此，建立准确度高、选择性好、检测限低、灵敏度高的检测方法将对人类医学研究有深刻意义。然而由于盐酸二甲双胍紫外吸收不明显，导致直接测定时灵敏度较低。本论文首次采用对氯苯甲酰氯为衍生化试剂，对盐酸二甲双胍进行柱前衍生化反应，通过实验，得到最佳实验条件为：检测波长249.0nm；最佳流动相甲醇∶缓冲（KH₂PO₄+H₃PO₄，pH=4）=80∶20；流速0.8mL／min；柱温25℃；进样量20μL；最佳溶剂为二氯甲烷；最佳氢氧化钠浓度为2.5mol／L；最佳反应时间为60min；最佳投料比为4∶1；反应温度为室温。在以上最佳实验条件下，测得标准曲线的线性范围为0.0050mg／L～50.0mg／L，最小检测浓度为5.0μg／L，三种不同浓度二甲双胍标准液0.20mg／L、5.0mg／L、20.0mg／L日内精密度和日间精密度RSD分别为2.81％、0.92％、0.47％和3.54％、3.32％、1.41％。该三种浓度下的回收率分别为98.5％～103.3％、96.13％～102.9％、93.93％～96.38％。通过以上研究发现，盐酸二甲双胍通过衍生化后，有明显的紫外特征吸收。本论文采用柱层析色谱分离法提纯衍生化产物，对提纯物进行紫外光谱法、红外光谱法、核磁共振法及质谱法鉴定，确定了盐酸二甲双胍衍生化产物的结构，并在此基础上研究确定了其衍生化机理。从产物结构来看，该衍生化反应与普通的氨基化合物衍生化反应有明显的不同，反应过程中盐酸二甲双胍的主结构发生了变化，成为环状，生成2-氨基-4-二甲氨基-6-对氯苯基-1，3，5三嗪。最后，本论文将该衍生化方法引入模拟血液样品，测定其中的盐酸二甲双胍含量。经对比得出血样的最佳前处理方法。模拟样品工作曲线的一元线性回归方程为：y=11.511x-0.1239，线性回归系数r=0.9985，线性范围为1.0mg／L～20.0mg／L，最低检测浓度为5.0μg／L。模拟血液样品2.0mg／L、5.0mg／L、20.0mg／L三种浓度的日内精密度和日间精密度RSD分别为5.40％、1.98％、3.22％和5.59％、2.52％、4.17％，该三种浓度下的回收率分别为94.92％～101.68％、98.23％～102.78％、96.88％～100.08％。更多还原

【Abstract】 Diabetes is a kind of chronic lifelong disease which is related to thepeople health. Metformin hydrochloride has become first choice oftreatment medicine by the diabetic, especially for the obese patients.Therefore, development a method with high selectivity, accuracy and lowdetection limit will have profound significance to the humanity medicalresearch. However, when determined directly the selectivity is not sosatisfactory because of low or non-absorption of metforminhydrochloride.This paper use p-chlorobenzoyl chloride as a derivatization reagentto react with metformin hydrochloride for the first time. By this way, thepre-column derivatization can be received. According to the experiment,the optimum experimental conditions are as follows: the wavelength is at249.0nm; methanol: buffer solution: （KH₂PO₄+H₃PO₄, pH=4）=80:20;the flow rate is 0.8mL/min; the column temperature is at 25℃; thesamples size is 20μL; the optimum solvent is dichloridethane; theoptimum concentration of NaOH is 2.5mol/L; the optimum reaction time is 60min; the optimum throw material of than is 4:1; the optimumtemperature is room temperature. Under the optimum experimentalconditions, the linear range of calibration curve is 0.0050mg/L～50.0mg/L, the minimum detecting concentration is 5.0μg/L, the within-dayand inter-day precision RSD of 0.20mg/L、5.0 mg/L、20.0 mg/L are2.81％、0.92％、0.47％and 3.54％、3.32％、1.41％respectively. Therecovery are 98.5％～103.3％、96.13％～102.9％、93.93％～96.38％respectively.According to the experiment, the metformin hydrochloride has aclear ultraviolet absorption after derivation. A method named columnchromatography chromatography is used to purify the derivatizationproduct. Then UV、IR、NMR and MS are used to identified the structure.The mechanism of the reaction is also be studied. After reaction,the main structure of metformin hydrochloride is changed. Themechanism of the reaction is also be studied.Finally, the method is used to determine the simulation blood sample.Under the optimum preprocessing, the linear range of calibration curve is1.0mg/L～20.0mg/L, the minimum detecting concentration is 5.0μg/L,the within-day and inter-day precision RSD of 2.0 mg/L、5.0 mg/L、20.0mg/L are 5.40％、1.98％、3.22％and 5.59％、2.52％、4.17％respectively.The recovery are 94.92％～101.68％、98.23％～102.78％、96.88％～100.08％respectively.更多还原