毛细管电泳电化学发光检测用于生物分析的两个实例-分析化学专业毕业论文.docxVIP

山东夫学颂L学位论文为7 山东夫学颂L学位论文 为7 78 pM。 Km随维牛素H浓度升高而降低，而Km／Vmax的值基本不变· 说明维牛素H对PLD的抑制为反竞争性抑制。而扑热息痛对脯氨酸肽酶的抑 制为非竞争性抑制，扑热息痛对脯氨酸肽酶的抑制常数为9．73x103 pg／mL． 关键词：毛细管电泳，电化学发光，脯氨酸肽酶，糖尿病，药物。 n 山东大学硕士学位论文ABSlRACT 山东大学硕士学位论文 ABSlRACT In chapter one of this paper，a simple and sensitive method for plasma prolidase activity assay was developed based on Capillary Electrophoresis combined with Electrochemiluminescence detection(CE—ECL)of the formed proline．At optimal conditions(pH 7．5，30 mmol／L phosphate buffer，5 mmol／L Ru(bpy)3”，potential of working electrode：1．3v,injection：1 0 kV,1 0s)，the detection limit of proline was 12．2 fmol(S／N=3)，corresponding t01．22x104 U of prodase catalyzing for I min．In experiments，the conditions of proline detection and the enzymatic reaction system were optimized．The optimal conditions of the enzymatic reaction were：(I)preincubation：plasma diluted with 3．O×1 0。mol／L(pH 7．61 phosphate buffer,37‘C for 24 h with the concentration of Mn2+was 1．0x 1 0‘3 tool／L；(2)incubation：Mn2+，I．0x 1 0’3 mol／L；glycylproline， 0．0 1 tool／L；37’C for 30 min；(3)termination：80‘C for l 0 min．The feproducibility(RSD，n=1 0)was 3．47％and the recoveries of five samples were between97．0％and 1 06％．The prolidase activity of 4 plasmas samples and 2 serum samples determined by CE-ECL was in agreement with that by classic Chinard’s method．The new method was also applied to detect PLD activity in serum of diabetic patients．MeanaSD was 888．1 g 4-2 1 i．48 U／L for the 45 serum samples studied which obey the normal distribution．There was a significant positive correlation between serum PLD activity and fasting serum glucose concentration in the 38 SUbjeets with normal serum triglyceride levels(r=O．495， p0．002)．Serum PLD activity in diabetes mellitus was higher than that in normal fasting glucose(pO．05)，which indicated that coll；,gen degradation was accelerated in diabetes mellitus．High triglyceride concentration maybe another factor th．at stimulate the disorder of collagen metabolism． In chapter two，