In this contribution a highly cost-effective flow analysis system for determination of alkaline phosphatase (ALP) activity is presented. This fully-mechanized bioanalytical system is based on economic solenoid micropumps and microvalves (powered and actuated by Arduino microcontroller) and extremely cheap dedicated optoelectronic flow-through detectors allowing absorbance and fluorescence measurements. The detection schemes for ALP assaying realized in this system are based on orthophosphate determination. For the detection of these ions formed in the course of enzymatic conversion a molybdate method requiring only common and inexpensive chemicals is utilized. Thus, for the enzymatic assays the simplest not-chromogenic/not-fluorogenic ALP substrates can be applied. Such approach results in the use of low-cost reagents for ALP assays, whereas the mechanization of assay causes low consumption of reagents as well as samples. In the course of reported investigations six ALP substrates were examined and the most promising results have been obtained for the inorganic compound – monofluorophosphate (MFP). The obtained linear ranges of absorbance and fluorescence measurements are 100–600 U L⁻¹ and 30-30/30-100 U L⁻¹, with sensitivities of 0.7 mV L⁻¹ U⁻¹ and 2.3/1.0 mV L⁻¹ U⁻¹, respectively. The calculated limits of detection are 5.1 U L⁻¹ (photometry) and 0.9 U L⁻¹ (fluorometry). The throughputs of the developed system are 13 and 12 samples/h for photometric and fluorometric detections, respectively. To demonstrate the practical utility of the developed bioanalytical system the ALP assays in complex matrix samples have been carried out. The results of ALP activity determination in serum samples are well-correlated with those obtained using reference method recommended for routine clinical analysis.

在这项贡献中，提出了一种用于测定碱性磷酸酶（ALP）活性的极具成本效益的流量分析系统。这种完全机械化的生物分析系统基于经济的螺线管微型泵和微型阀（由Arduino微控制器提供动力和驱动）以及极其便宜的专用光电流通检测器，可进行吸光度和荧光测量。在该系统中实现的用于ALP测定的检测方案基于正磷酸盐测定。为了检测在酶促转化过程中形成的这些离子，使用了仅需要普通且廉价的化学品的钼酸盐方法。因此，对于酶促测定，可以使用最简单的不发色/不发荧光的ALP底物。这种方法导致将低成本试剂用于ALP分析，而测定的机械化导致试剂和样品的低消耗。在报告的研究过程中，检查了六种ALP底物，无机化合物一氟磷酸盐（MFP）获得了最有希望的结果。所获得的吸光度和荧光测量值的线性范围为100–600 U L^-1和30-30 / 30-100 U L ^-1，灵敏度分别为0.7 mV L ^-1 U ^-1和2.3 / 1.0 mV L ^-1 U ^-1。计算的检测极限是5.1 U L ^-1（光度法）和0.9 U L ^-1（荧光法）。对于光度和荧光检测，开发的系统的吞吐率分别为13和12个样本/小时。为了证明所开发的生物分析系统的实用性，已经在复杂基质样品中进行了ALP分析。血清样品中ALP活性测定的结果与使用推荐用于常规临床分析的参考方法获得的结果密切相关。