【作者】 冯书惠；

【导师】 屈凌波； 李建军；

【作者基本信息】 郑州大学 ， 分析化学， 2003， 硕士

【摘要】 近年来HPLC在我国的发展突飞猛进，并且此发展势头还在继续增长，在21世纪，HPLC已发展成为使用频率很高的一种仪器分析方法。在药物分析中，HPLC一直是研究体内药物及其代谢物最强有力的手段。药物分析发展的主要趋向就是如何能够简便、快速地从复杂组成的样品中灵敏、可靠地检测一些微量成分，研究机体对药物的处置过程，特别是研究血浆药物浓度随时间而变化的数量规律。此外，疾病诊断和和滥用药物的检测也是药物分析的重要领域。 本文的研究内容主要包括四个方面：1 以高效液相色谱法测定新药莫索尼定胶囊中有效成分为指标，建立了检测莫索尼定的方法，并考察了强光、高温、中温、湿度对莫索尼定胶囊稳定性的影响，色谱柱ODS（200mm×4.6mm；）；流动相：甲醇：水：四氢呋喃（55：45：1）；流速：0.7mL／min；检测波长：254nm。结果表明莫索尼定胶囊在低于120℃、强光、潮湿条件下其主成分都能保持恒定不变，具有很好的稳定性。2 建立了反相高效液相色谱-紫外检测法检测盐酸曲马多浓度的方法，并对其体内代谢动力学进行了研究。血样预处理采用两次液-液反萃法。采用ODS色谱柱（150×4.6mm）；流动相为乙腈：磷酸盐缓冲溶液（0.02mol／L，pH3.7）=17：83（V／V）；流速：1.0mL／min；检测波长：λ=216nm。血浆中盐酸曲马多的最低检测浓度为10.2ng／mL，在25～800ng／mL的浓度范围内线性关系良好；曲马多在健康人体内呈现一级吸收二室开放模型特点。3 研究了高效液相色谱法测定血浆中二甲胺四环素的各种条件，并对其体内代谢动力学进行了研究。采用C₁₈反相色谱柱。流动相为：0.2mol／L 草酸胺：二甲基四胺：0.1mol／LEDTA（550：250：200，V：V：V），用0.1mol／L氢氧化四丁基胺调pH 6.2，紫外检测，λ=355nm，线性范围为1～8μg／mL，检出限为0.2ng，日间和日内变异系数小于4.0％；二甲胺四环素在健康人体内呈现一级吸收二室开放模型特点。4 在疾病诊断方面的工作：苯丙酮尿症（PKU）检查常用的方法很多，如聚合酶链反应技术诊断法、基因诊断法、温度梯度聚丙烯凝胶电泳诊断法、聚酰胺薄膜荧光扫描法等，其中以Guthrie氏细菌抑制法最为方便，具有采血少、特异性强、敏感性较高，对新生儿干扰和伤害小的优点，易取得家长或婴儿室的合作，而且操作要求不高，手续简便，不需特殊设备，极易进行大规模筛查。本文依据微生物法与化学法相结合，运用苯丙氨酸与β-2-噻吩丙氨酸竞争抑制原理开发研制出了苯丙酮尿症（PKU）筛查试剂盒，并对187例临床新生儿血标本进行了筛查，结果与确诊结果相符，灵敏度为2mg／100mL，结果可靠，价格低廉，适合在广大医院进行新生儿血常规筛查使用。更多还原

【Abstract】 The application of HPLC in pharmacology advances very rapidly in our country in recent years, and the trend is still more and more powerful. It is forecasted to be one of the strongest analysis means in 21st century, especially in clinical parmaco-kinetics.The dissertation mainly consists of four aspects. 1 The stabilityof moxonidine capsule was studied, and the content of moxonidine capsule was determined by means of RP- HPLC. The samples were tested by accelerating experiment % room temperature storage and strong light exposure experiment respectively. Under the normal conditions moxonidine capsule is very stable , but when temperature is above 120#, moxonidine is decomposed, and the stability of capsule is destroyed. 2 A reversed-phase high performance liquid chromatography (HPLC) method was established to determine the concentration of tramadol hydrochloride in human plasma, and studied its pharmaco-kinetics. HPLC instrument was used with the column:150mm × 4. 6mm(Lichrosorb C18, 5μm). The mobile phase was composed of phosphate buffer solution (0. 02mol/L, pH=3. 7)-acetonitrile (83:17,V/V). Flow rate was 1.0mL/min. Detection wavelength was 216nm. The standard curve equation was Y=0.03244X+0. 6007, The linear range was 25~800ng/mL. The minimum detection limit was 10. 2ng/mL. Pharmaco-kinetics parameter for tramadol hydrochloride tablet were T1/2Ka=0.45±0.21h, T1/2 beta =5.85±2.48h, Tpeak=1.65 ±0. 52h,Cmax=348.60±98.65ng/mL The area under the curve of concentration versus time after 100mg oral of tramadol hydrochloride tablet was AUC0-=3245. 6±1516. 02(ng/mL).h. The reversed-phase HPLC method is simple, rapid and sensitive. It is applicable to determine the concentration oftramadol hydrochloride in human plasma. 3 Conditions have been studied forthe determination of minocycline in human plasma by RP-HPLC. Chromatographic separation has been achieved on C18 column with 0. 2mol/Loxalic amine: bimethyl-tetramine: 0.1mol/LEDTA(550:250:200) as the mobile phase. Detection was at UV 355nm. The drug and an internal standard were extracted in ethyl-acetate from buffered plasma(pH6.2). The standard curve of minocycline was linear in the range of 1?μg/mL. The minimum detection concentration was 0.2ng. Pharmaco-kinetics parameter for minocycline tablet were Tpeak=2.69±0.9111, (Cmax,=4. 45±0.95μg/mL. The area under the curve of concentration versus time after 100mg oral of minocycline tablet was AUC =32.45±15.16 (μg/mL). h. The reversed-phase HPLC method is simple, rapid and sensitive. It is applicable to determine the concentration of minocycline in human plasma.Moreover, we also have done some work on developing phenylketonuria (PKU) screening kit. The clinical experiments validated that it was simple, sensitive and stable. It is very convenient to extend in many common hospitals.更多还原