The invention relates to fluorescent probe detection, in particular to a derivative L of rhodamine B, preparation thereof and use thereof. The derivative L of the rhodamine B has a structural formula (I). The preparation method of the derivative L of the rhodamine B comprises : firstly, reacting the product of the hydrazinolysis of the rhodamine B with glyoxal to obtain an intermediate compound; and secondly, reacting the intermediate compound with benzyldithiocarbazate in a molar ratio of 1 : 1 in absolute ethanol under a refluxing condition for 4 to 6 hours, cooling, filtering precipitated solid, washing with absolute ethanol and ethyl ether in turn and drying under vacuum to obtain a target compound. The derivative L of rhodamine B can serve as a fluorescent probe. In the invention, the derivative L of rhodamine B is obtained by an effective synthesis means, the copper ion selectivity is high, and the detection of copper ions can be realized on the basis of optimized experimental conditions.
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