血清总蛋白测定试剂的改良与应用效果评价

目的　探讨能消除脂质与葡聚糖双重干扰的血清总蛋白测定方法。方法　用氢氧化钾代替Doumas法试剂中的氢氧化钠,使脂质的溶解度增大,以消除或减低脂质所致反应液混浊。优选调整酒石酸钾钠的浓度,以消除葡聚糖对测定的干扰。用正交法设计实验,优选氢氧化钾和酒石酸钾钠的最佳浓度。结果　含氢氧化钾 143mmol/L、酒石酸钾钠 56.7mmol/L的改良试剂与含 TG 5.6mmol/L、葡聚糖60g/L的血清反应均不显混浊,可不设标本空白。TG>5.6mmol/L时,反应液混浊程度较Doumas法轻,可用标本空白纠正。对蛋白标准液和新鲜混合血清反应的吸收曲线相同。吸收峰530~560nm,最大峰值546nm,与Doumas法一致。改良试剂和Doumas试剂与蛋白反应的比吸光度分别为0.299 2L·g-1·cm-1和0.303 6L·g-1·cm-1。测定线性范围28~140g/L。回归方程y=0.892 5+0.272 7xL·g-1·cm-1,r=0.999 5。与 Doumas法平行测定外观正常的血清 72 例。配对 t检验,t=0.99,P>0.2。结论　改良试剂性能稳定,成本低廉,既能消除脂质与葡聚糖的双重干扰,又保留了Doumas法的优点。适合手工、半自动和全自动生化分析仪测定血清总蛋白。

The modified reaseach and effective evaluation of serum total protein's measure reagent

Objective To explore serum total protein's measure methods of eliminating lipoprotein and glucose polymer's double distraction.Methods Potassium hydroxide was used to replace reagent sodium hydroxide of Doumas method, which enhanced the solubility of lipoprotein and decreased or eliminated the liquid turbidity resulted from lipoprotein. Adjusting the thickness of tartrate kaliumna natrium was to eliminate the distraction of glucose polymer. Designing experiment through planning, we selected the best thickness of potassium hydroxid and tartrate kaliumna natrium.Results The reaction was not turbidness which was between modified reagent containing potassium hydroxide 143 mmol/L, tartrate kaliumna natrium 56.7 mmol/L and blood serum containing triglyceride 5.6 mmol/L,glucose polymer 60 g/L and needn't sample blank.When triglyceride >5.6 mmol/L, the liquid turbidity was lighter than Doumas method and can be adjusted by sample blank. The abstract curves of protein standard liquid and fresh mixture serum were same, which abstract peak value was 530-560nm,the maximum one was 546 nm according to Doumas method. The absorbency which the modified and Doumas reagent reacted with protein were respective 0.2992 and 0.303 6. The measure lineal limit was 28-140 g/L and regress equation was y=0.8925+0.2727x, r=0.9995. At the same time, we measured normal serum 72 cases and did t test, t=0.99, P>0.2.Conclusion Modified reagent has steady capability and lower price. It is not only eliminate double distraction of lipoprotein and glucose polymer but also reserve the advantage of Doumas method. It adapts to handicraft,half-automatic biochemistry analysis and automatic biochemistry analysis.Its effection of appliance is satisfactory.