Abstract

A new fluorometric method was developed for the determination of α-amylase activity in human serum samples. Firstly, a saturated starch–iodine complex (SI) was prepared. The SI complex was combined with sodium fluorescein to form a starch–iodine–sodium fluorescein complex (SIF). As the SIF complex decomposes with the α-amylase enzymatic hydrolysis of starch, the intensity of its fluorescence emission increases. The α-amylase activity is determined using the increased fluorescence emission intensity following hydrolysis of the SIF complex by α-amylase. The optimum pH, optimum buffer concentration, optimum temperature, and interference effect were identified for the developed fluorometric measurement method. Under the optimum conditions, a linear calibration curve was obtained between 0.18 and 9.00 U/L for α-amylase. The α-amylase activity in the human serum sample was also determined by our prepared measurement system and compared with the result from a medical center. Both methods are in good agreement with each other. Because this newly developed fluorometric method for α-amylase activity in serum samples is inexpensive, easy to use, and carried out to detect a very low amount of human serum α-amylase with sensitivity, it can be proposed this method for alpha-amylase activity assay in all other biological samples.

摘要

开发了一种新的荧光测定方法，用于测定人血清样品中的α-淀粉酶活性。首先，制备了饱和淀粉-碘复合物（SI）。SI复合物与荧光素钠结合形成淀粉-碘-荧光素钠复合物（SIF）。当SIF复合物随着淀粉的α-淀粉酶酶解而分解时，其荧光发射强度增加。在SIF复合物被α-淀粉酶水解后，使用增加的荧光发射强度来确定α-淀粉酶活性。为开发的荧光测量方法确定了最佳pH，最佳缓冲液浓度，最佳温度和干扰效应。在最佳条件下，α-淀粉酶的线性校准曲线在0.18和9.00 U / L之间。人血清样品中的α-淀粉酶活性也通过我们准备的测量系统确定，并与医疗中心的结果进行了比较。两种方法彼此非常吻合。因为这种新开发的用于血清样品中α-淀粉酶活性的荧光测定法价格低廉，易于使用且可用于检测极少量的人血清α-淀粉酶具有敏感性，因此可以建议将该方法用于所有其他生物样品中的α-淀粉酶活性测定。