Department of Chemistry, Forman Christian College (A Chartered University) , Lahore, Pakistan.
Department of Chemistry, School of Science and Technology, University of Management and Technology , Lahore, Pakistan.
Biosci Biotechnol Biochem. 2020 Oct;84(10):1967-1974. doi: 10.1080/09168451.2020.1781593. Epub 2020 Jul 3.
This work presents the development and validation of a simple, rapid, and cost-effective spectrophotometric method for quantitative analysis of uric acid in biological samples. The method relies upon uric acid-led reduction of Fe(III) to Fe(II) of sample/standard solutions which stoichiometrically engages ferrozine to form a magenta-colored complex. Different parameters including pH, metal and chelator concentrations, temperature, etc., were optimized for the maximum intensity and stability of the complex. The uric acid concentrations of synthetic/plasma solutions were determined by comparing the color intensity of Fe(ferrozine) complex produced by test solution with the standard curve formed by known uric acid concentrations. The method was validated in accordance with ICH guidelines and subjected to human plasma analysis. The results obtained were compared with a reference (enzymatic) method which revealed that there was no significant difference between the two methods at 95% confidence level. The method is highly specific, precise, linear, accurate, and robust.
本工作提出了一种简单、快速且经济有效的分光光度法，用于定量分析生物样品中的尿酸。该方法依赖于尿酸诱导的样品/标准溶液中 Fe(III)还原为 Fe(II)，然后化学计量地与菲啰嗪结合形成紫红色络合物。优化了不同参数，包括 pH 值、金属和螯合剂浓度、温度等，以获得络合物的最大强度和稳定性。通过将测试溶液产生的 Fe（菲啰嗪）络合物的颜色强度与已知尿酸浓度形成的标准曲线进行比较，确定合成/血浆溶液中的尿酸浓度。该方法按照 ICH 指南进行了验证，并进行了人血浆分析。所得结果与参考（酶）方法进行了比较，在 95%置信水平下，两种方法之间没有显著差异。该方法具有高度的特异性、精确性、线性、准确性和稳健性。
