The equivalent-circuit model (ECM) is widely used in online estimating the parameters and states of lithium-ion batteries. However, the sampling delay between the voltage and current of a battery is generally overlooked, which is unavoidable in a modular battery management system (BMS) and would lead to wrong results in the estimation of battery parameters and states. In this paper, with the first-order resistor–capacitor (RC) model as our battery model, we analyze the influence mechanism of sampling delay and then propose an optimized method for online estimating battery parameters. The mathematical model derived from the first-order RC model and the approximation method of first-order derivative are optimized. The recursive least squares (RLS) algorithm is used for identifying the parameters of the model. In order to verify the proposed method, a modular battery test system with high sampling frequency and high synchronization accuracy is developed. The experiment results indicate that the sampling delay would cause the estimation process to fluctuate, and the optimized method effectively improves the tolerance range of sampling delay.