HPLC analysis of a reference standard of neotame and sample isolates from flavoured milk was performed under the following set of standardized conditions: The HPLC system for neotame was the same as in the case of aspartame. The mobile phase was 0.09% TFA, water: acetonitrile (60:40) pumped at a flow rate of 0.6 ml/min. The column temperature was maintained at 40 ° C in a column heater chamber. The sample volume was 20 μl. A PDA detector at 210 nm was used to monitor the eluate.