Disposable electrochemiluminescent biosensor for lactate determination in saliva
Disposable electrochemiluminescent biosensor for lactate determination in saliva
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DOI：
10.1039/b821922b
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发表时间：
2009-01-01
期刊：
影响因子：
4.2
通讯作者：
Capitan-Vallvey, L. F.
中科院分区：
文献类型：
--
作者：
Ballesta Claver, J.;Valencia Miron, M. C.;Capitan-Vallvey, L. F.
研究了一种基于电化学发光的一次性乳酸生物传感器。乳酸识别系统以乳酸氧化酶(LOX)为基础，转导系统由鲁米诺组成。所有需要的试剂，鲁米诺，LOX，牛血清白蛋白，电解液和缓冲液已经被放置在丝网印刷电化学电池工作电极上的Metocel(TM)膜固定。当50亩L样品被放入装有一次性敏感膜的圆形容器的丝网印刷池中时，通过光计数头可以实现电化学发光的测量。对膜的组成和反应条件进行了优化，以获得足够的灵敏度。当施加两个0.5V的1 S脉冲时，一次性生物传感器在20 S后响应乳酸，以获得分析参数，即电化学发光初始速率。乳酸的线性双对数依赖关系的动态范围为10(5)~5×10(4)M，检出限为5×10(-6)M，传感器对传感器的重复性，在该范围的中等水平上的相对标准偏差RSD为3.30%。将ECL一次性生物传感器应用于唾液中乳酸的分析，作为一种非侵入性获取唾液中乳酸水平的替代方法。研究了来自唾液成分的干扰，并以一种易于处理的简单方式消除了干扰。该程序被验证用于人类唾液，并将结果与酶参考程序进行比较。该方法具有快速、廉价、选择性好、灵敏度高等优点，并使用常规的电化学发光仪器。
An electrochemiluminescence-based disposable biosensor for lactate is characterized. The lactate recognition system is based on lactate oxidase (LOx) and the transduction system consists of luminol. All the needed reagents, luminol, LOx, BSA, electrolyte and buffer have been immobilized by a Methocel (TM) membrane placed on the working electrode of the screen-printed electrochemical cell. The measurement of the electrochemiluminescence (ECL) is made possible via a photocounting head when 50 mu l of sample is placed into the screen-printed cell with a circular container containing the disposable sensing membrane. The compositions of the membrane and reaction conditions have been optimized to obtain adequate sensitivity. The disposable biosensor responds to lactate after 20 s when two 1 s pulses at 0.5 V are applied to obtain the analytical parameter, the ECL initial rate. The linearized double logarithmic dependence for lactate shows a dynamic range from 10 (5) to 5 x 10 (4) M with a detection limit of 5 x 10(-6) M and a sensor-to-sensor repeatability, as relative standard deviation, RSD, of 3.30% at the medium level of the range. The ECL disposable biosensor was applied to the analysis of lactate in human saliva as an alternative procedure for obtaining the lactate level in a non-invasive way. Interferences coming from components of saliva were studied and eliminated in a simple way that was easy to handle. The procedure was validated for use in human saliva, comparing the results against an enzymatic reference procedure. The proposed method is quick, inexpensive, selective and sensitive and uses conventional ECL instrumentation.