Simultaneous quantification of total carbohydrate and protein amounts from aqueous solutions by the sulfuric acid ultraviolet absorption method (SA-UV method).
Simultaneous quantification of total carbohydrate and protein amounts from aqueous solutions by the sulfuric acid ultraviolet absorption method (SA-UV method).
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DOI：
10.1016/j.aca.2021.338712
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发表时间：
2021-08
影响因子：
6.2
通讯作者：
Swen Ehnert;Jürgen Seehase;C. Müller‐Renno;M. Hannig;C. Ziegler
中科院分区：
文献类型：
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作者：
Swen Ehnert;Jürgen Seehase;C. Müller‐Renno;M. Hannig;C. Ziegler
基于硫酸紫外测定法（SA-UV，由Albasmeh等人开发，2013年），我们进一步扩展了该方法，用于通过紫外分光光度法同时定量糖类（碳水化合物）和蛋白质。糖类的吸光度取决于在浓硫酸存在下脱水形成糠醛，而蛋白质不受影响，并且可以通过紫外线活性肽键和芳香族氨基酸残基进行定量。在糖/蛋白质混合物中，SA-UV 测定提供了一种很好的替代方法，可以替代两种不同方法的需要，例如苯酚硫酸（PSA，由 DuBois 等人开发，1951 年）和二辛可宁酸（BCA，由 Smith 等人开发，1985 年）测定法。为了开发该方法，我们使用葡萄糖和 BSA 作为模型底物，并在线性、LOD、LOQ、准确性和精密度方面进行了方法验证。葡萄糖/BSA 混合物的同时定量可以从 30 µL 样品体积低至 20 mg/L，甚至浓度低至 2 mg/L 的低含量混合物也可以通过蒸发技术从更高的体积中适当地定量。
Based on the sulfuric acid-ultraviolet assay (SA-UV, developed by Albalasmeh et al., 2013), we have further expanded this method for the simultaneous quantification of saccharides (carbohydrates) and proteins by ultraviolet spectrophotometry. The absorbance of saccharides depends on the formation of furfurals by dehydration in the presence of concentrated sulfuric acid, whereas proteins are unaffected and can be quantified by UV active peptide bonds and aromatic amino acid residues. In saccharide/protein mixtures the SA-UV assay offers a good alternative and substitutes the need for two different methods, like the phenol-sulfuric acid (PSA, developed by DuBois et al., 1951) and bicinchoninic acid (BCA, developed by Smith et al., 1985) assays. For the development of this method, we used glucose and BSA as model substrates and performed a method validation in terms of linearity, LOD, LOQ, accuracy, and precision. Simultaneous quantification in glucose/BSA mixtures is possible down to 20 mg/L from 30 μL sample volumes, and even low content mixtures with concentrations down to 2 mg/L can appropriately be quantified from higher volumes by an evaporation technique.