Rapid Quantification of Luteolin in Peanut Shells by Fluorescence Digital Imaging of Reactions Spotted on Filter Paper
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Abstract:
A rapid luteolin quantification method based on fluorescence imaging of reagents spotted on filter paper was established in this study. The method was based on the ability of luteolin to significantly enhance the fluorescence intensity of Tb3+ in the presence of EDTA. Luteolin solution and detection reagent (including EDTA,Tb3+,NaOH) were spotted onto a paper chip, then red (R), green (G), and blue (B) fluorescence intensities were captured and read under UV light using a smartphone. Image exposure conditions, interfering substances, and luteolin concentration calculation methods were examined using this quantification system. Optimal fluorescence development conditions were as follows: add sequentially 0.4 μL1.0 mol/L NaOH solution, 1.0 μL 1.0×10-2 mol/L EDTA solution, 6 μL luteolin solution, and 0.75 μL 1.5 mmol/L Tb3+solution to the paper chip, then allow reaction to proceed for 5 min at room temperature. Under these conditions, luteolin will have a good linear relationship with B/(R+G+B) at concentrations between 0.05 and 4.0 mmol/L, R2=0.9954. When using this rapid quantification method to detect the luteolin content in peanut shells, recovery rates were between 96.5% and 106.3%, with a relative standard deviation of 3.9%. This method was rapid, simple, and economical, and allowed on-the-spot quantification of luteolin in complex samples.
