Characterizing the Existence of Fluorescence Quenching Agents Using EEM Fluorescence and UV Spectra: Taking the Interaction of Humic Acid and Fe(Ⅲ) as an Example
LI Wei-hua1, 2, WU Gun1, 2, YAO Liang1, 2, HUANG Xian-huai2, WANG Jia-qin1, 2, SHEN Hui-yan1, 2, XUE Tong-zhan1, 2
1. School of Environment and Energy Engineering, Anhui Jianzhu University, Hefei 230601, China 2. Key Laboratory of Water Pollution Control and Wastewater Reuse in Anhui Province, Anhui Jianzhu University, Hefei 230601, China
Abstract：The fluorescence quenching agents was characterized with three-dimensional fluorescence and ultraviolet (UV) spectra. When there was Fe (Ⅲ) in the sample, the humic fluorescence would be quenched and their UV spectra were not affected. The variation of fluorescence intensity (I) at Ex/Em=300/510 nm and UV absorbance(A) at UV300 were investigated in the article. The smaller the ratio of fluorescence intensity versus UV absorbance (I/A) is, the higher the fluorescence quencher Fe(Ⅲ) concentration is. According to Stern-Volmer equation I/I0=1-fc×Kc×[c] /(1+Kc×[c] ) and fitted function I/A=f×[k/(cFe3++c)+b] , the fitted fluorescent quenching constant Kc was ranged between 1.08 to 1.15, the ratio of bounded fluorophores versus total fluorophores, i.e. fc, was ranged between 1.10 to 1.14. The ratio of fluorescence intensity and absorbance of humic acid was fitted with Fe(Ⅲ) concentration and the constants were acquired as following: f=0.83～1.19, k=587.19～612.19, c=0.87～0.92, b=-87.09～-46.36. The correlation curve values were 0.99. The Stern-Volmer formula was used to describe the quenching effect of humic acid fluorescence by Fe (Ⅲ). However, due to the fact that the fluorescence intensity I0 without quencher was difficult to acquire during the analysis of practical samples, the fitted function between the ratio of I/A and Fe(Ⅲ) was used to reflect the quenching effect of Fe(Ⅲ) on the fluorescence of humic acid, which was based on the correlations between the fluorescence intensity I0 and ultraviolet absorbance A. The fitted formula was used to predict the iron ions concentration of the resin separated and concentrated samples from wastewater treatment plant and receiving waters. The predicted values were in good accordance with those determined with inductively coupled plasma atomic emission spectroscopy(ICP-AES) method when the iron ion concentration was above 0.4 mg·L-1, which could be used to ascertain the existence of fluorescence quenching agent and their corresponding concentration.
李卫华1, 2，伍 鲧1, 2，姚 亮1, 2，黄显怀2，王佳琴1, 2，申慧彦1, 2，薛同站1, 2 . 三维荧光与紫外光谱联合表征样品中猝灭剂赋存状态：以胡敏酸与Fe(Ⅲ)相互作用为例 [J]. 光谱学与光谱分析, 2017, 37(03): 783-787.
LI Wei-hua1, 2, WU Gun1, 2, YAO Liang1, 2, HUANG Xian-huai2, WANG Jia-qin1, 2, SHEN Hui-yan1, 2, XUE Tong-zhan1, 2 . Characterizing the Existence of Fluorescence Quenching Agents Using EEM Fluorescence and UV Spectra: Taking the Interaction of Humic Acid and Fe(Ⅲ) as an Example . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(03): 783-787.
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