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Highly sensitive analysis of the constituents in whole blood diluted with chemiluminescence-FIA using a bioreactor

使用生物反应器对化学发光-FIA 稀释的全血中的成分进行高灵敏度分析

基本信息

批准号：
10672176
负责人：
TABATA Masayoshi
金额：
$ 1.98万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

Highly sensitive analysis of the constituents (glucose, uric acid) in whole blood diluted was tried using FIA (flow injection analysis) system involving a chemiluminescence detector (photon counting type) and an immobilized enzyme column reactor, so that a lot of constituents in blood were able to be determined with blood volume level which a mosquito drew.1. Glucose determination-Beforehand, whole blood was diluted with physiological saline solution or distilled water, and plasma with distilled water to 5000〜400000-fold. Glucose in the diluted samples was determinated using the FIA-chemiluminescence system involving a pyranose oxidase column reactor (2x20 mm). The sample volume injected was 10 μ l. The use of a dispenser (Accu-drive, made in Jamany) and the preservation of distilled water in a glass container contributed to improve the reproducibility of glucose determination sharply. As the resuly, the reproducibility was satisfactory even the sample diluted to a hundred thousand-fold. The results of glucose concentration obtained by this method using whole blood diluted beforehand with distilled water or physiological saline solution to a hundred thousand-fold by a Accu-drive dispenser were compared with those obtained with the glucose oxidase-hydrogen peroxide electrode method. The calculated linear regression and correlation coefficient were y=0.916x-4.05 and r=0.986 for distilled water, and y=0.910x-3.00 and r=0.988 for physiological saline solution, indicating excellent correlation between the results of the two methods. In the case of plasma diluted with distilled water to a hundred thousand-fold, the calculated linear regression and correlation coefficient were y=0.979x-3.1 and r=0.987, indicating excellent agreement between the results of the two methods over a wide range of glucose concentrations.

本研究尝试了使用由化学发光检测器（光子计数型）和固定化酶柱反应器组成的FIA（流动注射分析）系统，对稀释后的全血中的成分（葡萄糖、尿酸）进行高灵敏度分析，从而能够以蚊子吸入的血容量水平测定血液中的多种成分。葡萄糖测定--全血用生理盐水或蒸馏水稀释，血浆用蒸馏水稀释至5000 ~ 400000倍。使用涉及吡喃糖氧化酶柱反应器（2x20 mm）的FIA-化学发光系统测定稀释样品中的葡萄糖。进样体积为10 μ l。使用分配器（Accu-drive，Jamany制造）和将蒸馏水保存在玻璃容器中有助于显著提高葡萄糖测定的重现性。结果表明，样品稀释至10万倍后，其重现性仍令人满意。将通过该方法获得的葡萄糖浓度结果与通过葡萄糖氧化酶-过氧化氢电极方法获得的结果进行比较，其中，所述方法使用预先用蒸馏水或生理盐水溶液稀释至十万倍的全血，通过Accu-drive分配器获得。蒸馏水的线性回归方程为y=0.916x-4.05，相关系数r=0.986;生理盐水的线性回归方程为y=0.910x-3.00，相关系数r=0.988，两种方法测定结果具有良好的相关性。在用蒸馏水稀释至十万倍的血浆的情况下，计算的线性回归和相关系数为y=0.979x-3.1和r=0.987，表明在较宽的葡萄糖浓度范围内，两种方法的结果之间具有极好的一致性。