| Summary: | Abstract Advantame is a novel sweetener, and 3‐hydroxy‐4‐methoxy benzal acrolein is an important intermediate to synthesize the sweetener. The aim of this study was to evaluate a new low‐cost method to purify 3‐hydroxy‐4‐methoxy benzal acrolein, and the crude intermediate was used as raw material. The intermediate was purified using a low‐pressure column chromatography with a C18 column, using a methanol‐water (6:4, v/v) at a flow rate of 6.0 ml/min, and the loading amount of the crude intermediate in solution was 10.0 mg in total. A method for the analysis of 3‐hydroxy‐4‐methoxy benzal acrolein was established using high‐performance liquid chromatography (HPLC). This method was validated in terms of its linearity, repeatability, accuracy, detection limit, and quantitation limit. The calibration curves of 3‐hydroxy‐4‐methoxy benzal acrolein were linear (r > .999) over a wide concentration range of 0.005–0.08 mg/ml, by comparing the ratio of signal to noise, and the detection limit was 5.0 ng/ml and the quantification limit was 15.0 ng/ml. Good repeatability was obtained (RSD < 2%, n = 6), and the recoveries calculated using mixed sample previously quantified ranged from 94.5% to 106.37%. As long as, this method has been successfully applied to the analysis of 3‐hydroxy‐4‐methoxy benzal acrolein; therefore, the method can be put into practical use during the industrial synthesis and real‐time detection of the intermediate.
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