采用分光光度顺序注射法连续测定尿蛋白和葡萄糖。

Successive determination of urinary protein and glucose using spectrophotometric sequential injection method.

作者信息

Kanchana Watla-iad, Sakai Tadao, Teshima Norio, Katoh Shuji, Grudpan Kate

机构信息

Department of Applied Chemistry, Aichi Institute of Technology, 1247 Yachigusa, Yakusa-cho, Toyota 470-0392, Japan.

出版信息

Anal Chim Acta. 2007 Dec 5;604(2):139-46. doi: 10.1016/j.aca.2007.10.010. Epub 2007 Oct 13.

DOI:10.1016/j.aca.2007.10.010

PMID:17996535

Abstract

A new sequential injection (SI) system with spectrophotometric detections has been developed for successive determination of protein and glucose. The protein assay is based on ion-association of protein with tetrabromophenolphthalein ethyl ester (TBPE) in the presence of Triton X-100 at pH 3.2. The blue product is monitored for absorbance at 607 nm. For glucose, hydrogen peroxide, generated by the oxidation of glucose in the presence of glucose oxidase immobilized on glass beads packed in a minicolumn, is monitored using iron-catalyzed oxidation reaction of p-anisidine to form a red colored product (520 nm). The SI procedure takes advantage in performing the protein assay during the incubation period for glucose oxidation. Linear ranges were up to 10 mg dL(-1) human serum albumin (HSA) with a limit of detection (LOD) (3sigma) of 0.3 mg dL(-1), and up to 12.5 mg dL(-1) glucose with LOD of 0.08 mg dL(-1). R.S.D.s (n=11) were 2.7% and 2.5% (for 1 mg dL(-1) and 5 mg dL(-1) HSA) and 1.4% (9 mg dL(-1) glucose). Sample throughput for the whole assay of both protein and glucose is 6 h(-1). The automated system has been demonstrated for the successive assay of protein and glucose in urine samples taken from diabetic disease patients, with good agreement with the other methods. This developed SI system is an alternative automation for screening for diabetic diagnosis.

摘要

已开发出一种带有分光光度检测的新型顺序注射（SI）系统，用于连续测定蛋白质和葡萄糖。蛋白质测定基于在pH 3.2的Triton X-100存在下，蛋白质与四溴酚酞乙酯（TBPE）的离子缔合。监测蓝色产物在607 nm处的吸光度。对于葡萄糖，通过固定在填充于微型柱中的玻璃珠上的葡萄糖氧化酶存在下葡萄糖氧化产生的过氧化氢，利用对茴香胺的铁催化氧化反应形成红色产物（520 nm）进行监测。SI程序利用了在葡萄糖氧化的孵育期进行蛋白质测定的优势。线性范围高达10 mg dL⁻¹人血清白蛋白（HSA），检测限（LOD）（3σ）为0.3 mg dL⁻¹，高达12.5 mg dL⁻¹葡萄糖，LOD为0.08 mg dL⁻¹。相对标准偏差（R.S.D.s）（n = 11）对于1 mg dL⁻¹和5 mg dL⁻¹ HSA分别为2.7%和2.5%，对于9 mg dL⁻¹葡萄糖为1.4%。蛋白质和葡萄糖的整个测定的样品通量为6 h⁻¹。该自动化系统已用于对糖尿病患者尿液样本中的蛋白质和葡萄糖进行连续测定，与其他方法具有良好的一致性。这种开发的SI系统是糖尿病诊断筛查的一种替代自动化方法。