Three different mobile phases were chosen for the elution of
neotame viz. 0.09% TFA:water:acetonitrile (70:30, 80:20, 60:40).
0.09% TFA:water:acetonitrile (60:40) gave the best resolution as
compared to the other two mobile phases. Three flow rates (0.6,
0.8 and 1.0 ml/min) of the mobile phase were assayed for the estimation
of neotame. The neotame peak merged with the product
matrix peak at a flow rate of 0.8 and 1.0 ml/min, hence, a flow rate
0.6 ml/min was selected for the analysis. At this flow rate the
retention time increased and better resolution was obtained as
compared to the flow rate of 0.8 and 1.0 ml/min.