CN101851665B - 酶法测定1,5-脱水山梨醇试剂盒 - Google Patents

酶法测定1,5-脱水山梨醇试剂盒 Download PDF

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CN101851665B
CN101851665B CN2009100717259A CN200910071725A CN101851665B CN 101851665 B CN101851665 B CN 101851665B CN 2009100717259 A CN2009100717259 A CN 2009100717259A CN 200910071725 A CN200910071725 A CN 200910071725A CN 101851665 B CN101851665 B CN 101851665B
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adp
reagent
nadp
anhydro
glucitol
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CN101851665A (zh
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贺丽雅
张颖
于永光
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Harbin Engineering University
Harbin Medical University
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Harbin Medical University
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Abstract

酶法测定1，5-脱水山梨醇试剂，目前，测定操作费事难以批量检测，并且这些方法只能现配现用，不能长期保存。本发明的组成包括：水、3-羟-甲基氨基甲烷、氧化型辅酶II(NADP+)、海藻糖、2-甲基-4-异噻唑-3-酮、吐温-80、ADP依赖性己糖激酶(ADP-HK)、二磷酸腺苷(ADP)、AG-6-磷酸脱氢酶，各组份含量为1升水中含3-羟甲基氨基甲烷60-140毫摩尔，氧化型辅酶II(NADP+)2.26-2.38毫摩尔，海藻糖1-5克，2-甲基-4-异噻唑-3-酮0.1-0.6活性单位，吐温-800.1-0.6克，ADP依赖性己糖激酶(ADP-HK)1000～3000U，AG-6-磷酸脱氢酶1000～80000U，二磷酸腺苷(ADP)0.1-0.3毫摩尔。本发明用于测定1，5-脱水山梨醇。

Description

酶法测定1,5-脱水山梨醇试剂盒
技术领域：
本发明涉及一种酶法测定1，5-脱水山梨醇试剂。
背景技术：
糖尿病临床常见病，多发病，原因是胰岛素分泌不足引起，血浆中AG水平与血浆葡萄糖、HbAlc和果糖胺(FMN)呈显著负相关，有资料表明，血浆葡萄糖＞8.3mmol/L者，无一例伴有高水平的血浆AG[12]。糖尿病人的血浆AG显著抵于健康人，1型糖尿病人较之2型尤甚，新发病和久病的1型糖尿病人，血浆AG一般都在1umol/L以下[13]，甚至降低到不能测出。测定1，5-脱水山梨醇有高效液项色谱、微柱层析法、化学发光法等。这些方法比较准确，但操作费事难以批量检测，并且这些方法只能现配现用，不能长期保存。
发明内容：
本发明的目的是提供一种酶法测定1，5-脱水山梨醇试剂，具有特异性高，准确、微量、快速，不需要对样品预先处理，操作简单，并且可冷藏保存一年以上，长期使用。
上述的目的通过以下的技术方案实现：
一种酶法测定1，5-脱水山梨醇试剂，其组成包括：水、3-羟-甲基氨基甲烷、氧化型辅酶II(NADP+)、海藻糖、2-甲基-4-异噻唑-3-酮、吐温-80、ADP依赖性己糖激酶(ADP-HK)、二磷酸腺苷(ADP)、AG-6-磷酸脱氢酶，各组份含量为1升水中含3-羟甲基氨基甲烷60-140毫摩尔，氧化型辅酶II(NADP+)2.26-2.38毫摩尔，海藻糖1-5克，2-甲基-4-异噻唑-3-酮0.1-0.6活性单位，吐温-80 0.1-0.6克，ADP依赖性己糖激酶(ADP-HK)1000∽3000U，AG-6-磷酸脱氢酶1000∽80000U，二磷酸腺苷(ADP)0.1-0.3毫摩尔。
所述的酶法测定1，5-脱水山梨醇试剂，PH为8.0-10.0。
本发明的有益效果：
1.本发明配方中3-羟-甲基氨基甲烷为缓冲剂有维持PH的作用，吐温-80有乳化作用，ADP依赖性己糖激酶(ADP-HK)、AG-6-磷酸脱氢酶起催化剂作用、2-甲基-4-异噻唑-3-酮起防腐作用，保护剂海藻糖糖对ADP依赖性己糖激酶(ADP-HK)、AG-6-磷酸脱氢酶有保护作用、氧化型辅酶II(NADP+)作为底物反应，试剂抗干扰能力强，中等程度溶血，乳糜血，血糖20mmOl/l以下，胆红素100umol/l以下对测定不产生干扰，本试剂盒为液体试剂，有效期一年。冷藏一年试剂盒中酶活力损失小于5％。
2.本试剂比原来试剂的优越性是省去了冷冻干燥工艺，降低成本，使用方便。
具体实施方式：
实施例1：
取超纯水一升用前将水加热至沸，密封备用，将3-羟-甲基氨基甲烷60毫摩尔加入水中，调PH至8.5取氧化型辅酶II(NADP+)2.26毫摩尔、保护剂海藻糖1克、2-甲基-4-异噻唑-3-酮0.1活性单位、吐温-80 0.1克、ADP依赖性己糖激酶(ADP-HK)1000U、AG-6-磷酸脱氢酶1000U、二磷酸腺苷(ADP)0.1毫摩尔加入试剂，将所得的试剂溶液分装成10毫升/瓶，冷藏随时备用。
实施例2：
取超纯水一升用前将水加热至沸，密封备用，将3-羟-甲基氨基甲烷140毫摩尔，调PH至9.5取氧化型辅酶II(NADP+)2.38毫摩尔、保护剂海藻糖5克、2-甲基-4-异噻唑-3-酮0.6活性单位、吐温-80 0.6克、DP依赖性己糖激酶(ADP-HK)3000U、AG-6-磷酸脱氢酶80000U、二磷酸腺苷(ADP)0.3毫摩尔加入试剂，将所得的试剂溶液分装成5毫升/瓶，冷藏随时备用。
实施例3：
取超纯水一升用前将水加热至沸，密封备用。将3-羟-甲基氨基甲烷100毫摩尔，调PH至9.5取氧化型辅酶II(NADP+)2.30毫摩尔、保护剂海藻糖3克、2-甲基-4-异噻唑-3-酮0.5活性单位、吐温-80 0.3克、ADP依赖性己糖激酶(ADP-HK)2000U、AG-6-磷酸脱氢酶50000U、二磷酸腺苷(ADP)0.2毫摩尔加入试剂，将所得的试剂溶液分装成5毫升/瓶，冷藏随时备用。

Claims (1)

1.一种酶法测定1，5-脱水山梨醇试剂，其特征是：其由水、3-羟-甲基氨基甲烷、氧化型辅酶Ⅱ（NADP+）、海藻糖、2-甲基-4-异噻唑-3-酮、吐温-80、ADP 依赖性己糖激酶（ADP-HK）、二磷酸腺苷（ADP）、AG-6- 磷酸脱氢酶组成，各组份含量为1升水中含3-羟甲基氨基甲烷60-140毫摩尔，氧化型辅酶Ⅱ（NADP+）2.26-2.38毫摩尔，海藻糖1-5克, 2-甲基-4-异噻唑-3-酮0.1-0.6活性单位，吐温-80 0.1-0.6克，ADP 依赖性己糖激酶（ ADP-HK ）1000∽3000U，AG-6- 磷酸脱氢酶1000∽80000U，二磷酸腺苷（ ADP ）0.1-0.3毫摩尔，pH为8.0-10.0。
CN2009100717259A 2009-04-03 2009-04-03 酶法测定1,5-脱水山梨醇试剂盒 Active CN101851665B (zh)

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CN102175670B (zh) * 2010-12-30 2013-03-20 北京九强生物技术股份有限公司 血液1,5-脱水葡萄糖醇的检测方法及试剂盒
CN107703071B (zh) * 2017-09-11 2021-02-26 三诺生物传感股份有限公司 一种检测1,5-ag的试剂盒及方法

Citations (2)

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EP1213357A2 (en) * 2000-10-11 2002-06-12 Kyowa Medex Co., Ltd. Method and reagent for quantitative determination of 1, 5-Anhydroglucitol

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