CN105954526B - 一种胰岛素的酶免疫测定方法 - Google Patents

一种胰岛素的酶免疫测定方法 Download PDF

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CN105954526B
CN105954526B CN201610263735.2A CN201610263735A CN105954526B CN 105954526 B CN105954526 B CN 105954526B CN 201610263735 A CN201610263735 A CN 201610263735A CN 105954526 B CN105954526 B CN 105954526B
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曹朝晖
胡小波
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Abstract

本发明公开了一种胰岛素的酶免疫测定方法,包括胰岛素溶液、与胰岛素溶液反应的葡萄糖溶液、氧化剂溶液、盐酸溶液、酸式滴定管和盛放混合液的容量瓶,胰岛素的酶免疫测定方法包括以下步骤:溶液混合;由碘溶液得出剩余葡萄糖溶液含量,进而得出葡萄糖消耗的量,再根据葡萄糖和胰岛素的化学反应得出胰岛素的含量。该胰岛素的酶免疫测定方法检测配方简单,制作成本低,便于推广使用;根据葡萄糖消耗含量,根据其之间化学反应,能简便有效地对血液中胰岛素的浓度进行测量。

Description

一种胰岛素的酶免疫测定方法
技术领域
本发明涉及酶免疫测定技术领域,尤其涉及一种胰岛素的酶免疫测定方法。
背景技术
胰岛素对物质代谢的作用非常重要,总的效果是促进合成代谢,抑制分解代谢。若缺少胰岛素,则血糖升高。因此,测定血液中胰岛素浓度和血糖值对诊断、治疗糖尿病是很重要的。
发明内容
本发明的目的在于提供一种胰岛素的酶免疫测定方法,以解决上述背景技术中提出的问题。
为实现上述目的,本发明提供如下技术方案:一种胰岛素的酶免疫测定方法,其特征在于:包括胰岛素溶液、与胰岛素溶液反应的葡萄糖溶液、纯碘溶液、氢氧化钠溶液、盐酸溶液、酸式滴定管和盛放混合液的容量瓶。
优选的,该胰岛素的酶免疫测定方法包括以下步骤:
S1:选取纯度较高的一定量的胰岛素溶液、过量的葡萄糖溶液静置备用,
S2:按容量百分比配备葡萄糖溶液并分别放入容量瓶内:0.5摩尔浓度葡萄糖溶液:1份;1摩尔浓度葡萄糖溶液:1份、1.5摩尔浓度葡萄糖溶液:1份、2摩尔浓度葡萄糖溶液:1份、2.5摩尔浓度葡萄糖溶液:1份和3摩尔浓度葡萄糖溶液:1份;
S3:将静置备好的胰岛素溶液分成等量6份,然后分别加入到0.5摩尔浓度葡萄糖溶液:1份、1摩尔浓度葡萄糖溶液:1份、1.5摩尔浓度葡萄糖溶液:1份、2摩尔浓度葡萄糖溶液:1份、2.5摩尔浓度葡萄糖溶液:1份和3摩尔浓度葡萄糖溶液:1份中,用酸式滴定管取适量盐酸溶液6份,分别放入上述6组混合溶液缓缓搅拌15min,将6组混合放入到水浴锅中煮沸8min,冷却后,混合溶液均匀摇晃100-150次重复三次C6H12O6+I2+3NaOH=C5H11O5COONa+2NaI+2H2O②,2I+2NaOH=NaIO+NaI+H2O③,NaIO+NaI+2HCl=I2+2NaCl+H2O④;
S4:根据步骤S3中的反应式④盐酸溶液(HCl)的消耗分别得出6组混合液中碘溶液(I2)的剩余量进而得出碘溶液(I2)的消耗量,由反应式②碘溶液(I2)的消耗量得到反应式①中葡萄糖的剩余量;
S5:根据步骤S4中,由加入总的葡萄糖量减去由反应式②得出葡萄糖剩余量得到胰岛素消耗葡萄糖量,由胰岛素消耗葡萄糖量,由反应式①得出RNH2的含量,进而计算出①胰岛素含量。
与现有技术相比,本发明的有益效果是:本发明提供的一种胰岛素的酶免疫测定方法,该胰岛素的酶免疫测定方法简单,制作成本低,便于推广使用;根据葡萄糖消耗含量,葡萄糖与胰岛素的氨的衍生物之间化学反应,能简便有效地对血液中胰岛素的浓度进行计算。
附图说明
图1为本发明流程示意图。
具体实施方式
下面将结合本发明实施例,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
请参阅图1,本发明提供一种技术方案:一种胰岛素的酶免疫测定方法,其特征在于:包括胰岛素溶液、与胰岛素溶液反应的葡萄糖溶液、纯碘溶液、氢氧化钠溶液、盐酸溶液、酸式滴定管和盛放混合液的容量瓶。
该胰岛素的酶免疫测定方法包括以下步骤:
S1:选取纯度较高的一定量的胰岛素溶液、过量的葡萄糖溶液静置备用,
S2:按容量百分比配备葡萄糖溶液并分别放入容量瓶内:0.5摩尔浓度葡萄糖溶液:1份;1摩尔浓度葡萄糖溶液:1份、1.5摩尔浓度葡萄糖溶液:1份、2摩尔浓度葡萄糖溶液:1份、2.5摩尔浓度葡萄糖溶液:1份和3摩尔浓度葡萄糖溶液:1份;
S3:将静置备好的胰岛素溶液分成等量6份,然后分别加入到0.5摩尔浓度葡萄糖溶液:1份、1摩尔浓度葡萄糖溶液:1份、1.5摩尔浓度葡萄糖溶液:1份、2摩尔浓度葡萄糖溶液:1份、2.5摩尔浓度葡萄糖溶液:1份和3摩尔浓度葡萄糖溶液:1份中,用酸式滴定管取适量盐酸溶液6份,分别放入上述6组混合溶液缓缓搅拌15min,将6组混合放入到水浴锅中煮沸8min,冷却后,混合溶液均匀摇晃100-150次重复三次C6H12O6+I2+3NaOH=C5H11O5COONa+2NaI+2H2O②,2I+2NaOH=NaIO+NaI+H2O③,NaIO+NaI+2HCl=I2+2NaCl+H2O④;
S4:根据步骤S3中的反应式④盐酸溶液(HCl)的消耗分别得出6组混合液中碘溶液(I2)的剩余量进而得出碘溶液(I2)的消耗量,由反应式②碘溶液(I2)的消耗量得到反应式①中葡萄糖的剩余量;
S5:根据步骤S4中,由加入总的葡萄糖量减去由反应式②得出葡萄糖剩余量得到胰岛素消耗葡萄糖量,由胰岛素消耗葡萄糖量,由反应式①得出RNH2的含量,进而计算出①胰岛素含量。
根据④式滴定消耗标准盐酸(HCl)的量得出④式碘,④式碘与③式碘相同,得出③式碘消耗量,根据②式和所准备的碘溶液一定量的过量纯碘可得出①式中反应剩余葡萄糖量,再由所准备的葡萄糖总量减去①式中反应剩余葡萄糖量计算出①式中消耗的葡萄糖量,进而再由①式中消耗的葡萄糖量得出氨基衍生物的量。
尽管已经示出和描述了本发明的实施例,对于本领域的普通技术人员而言,可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由所附权利要求及其等同物限定。

Claims (1)

1.一种胰岛素的测定方法,其特征在于:包括胰岛素溶液、与胰岛素溶液反应的葡萄糖溶液、纯碘溶液、氢氧化钠溶液、盐酸溶液、酸式滴定管和盛放混合液的容量瓶;
包括以下步骤:
S1:选取纯度较高的一定量的胰岛素溶液、过量的葡萄糖溶液静置备用,
S2:按容量百分比配备葡萄糖溶液并分别放入容量瓶内:0.5摩尔浓度葡萄糖溶液:1份;1摩尔浓度葡萄糖溶液:1份;1.5摩尔浓度葡萄糖溶液:1份;2摩尔浓度葡萄糖溶液:1份;2.5摩尔浓度葡萄糖溶液:1份和3摩尔浓度葡萄糖溶液:1份;
S3:将静置备好的胰岛素溶液分成等量6份,然后分别加入到0.5摩尔浓度葡萄糖溶液:1份、1摩尔浓度葡萄糖溶液:1份、1.5摩尔浓度葡萄糖溶液:1份、2摩尔浓度葡萄糖溶液:1份、2.5摩尔浓度葡萄糖溶液:1份和3摩尔浓度葡萄糖溶液:1份中,用酸式滴定管取适量盐酸溶液6份,分别放入上述6组混合溶液缓缓搅拌15min,将6组混合放入到水浴锅中煮沸8min,冷却后,混合溶液均匀摇晃100-150次重复三次C6H12O6+I2+3NaOH=C5H11O5COONa+2NaI+2H2O②,I2+2NaOH=NaIO+NaI+H2O③,NaIO+NaI+2HCl=I2+2NaCl+H2O④;
S4:根据步骤S3中的反应式④盐酸溶液(HCl)的消耗分别得出6组混合液中碘溶液(I2)的剩余量进而得出碘溶液(I2)的消耗量,由反应式②碘溶液(I2)的消耗量得到反应式①中葡萄糖的剩余量;
S5:根据步骤S4中,由加入总的葡萄糖量减去由反应式②得出葡萄糖剩余量得到胰岛素消耗葡萄糖量,由胰岛素消耗葡萄糖量,由反应式①得出RNH2的含量,进而计算出反应式①胰岛素含量。
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CN1729016A (zh) * 2002-10-22 2006-02-01 瓦拉塔药品公司 糖尿病的治疗

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