Abstract: Disassembly of hydrogen bonded layer-by-layer (LbL) film, composed of poly(acrylic acid) (PAA) and poly(2-ethyl-2-oxazoline) (PEOX), is studied by an electrochemical stimulus. Poly(2-oxazoline)s have been widely studied for biomedical applications due to their biocompatibility, thermal responsive behavior and easy chemical modification. Nowadays, one emerging research area for the development of biomedicine and gene engineer focuses on the controlled dissolution of LbL assemblies. A new method to trigger the hydrogen bonded LbL film dissolution near the positive electrode in the electrolytic bath is reported, which can obtain a uniform and transparent film. Through the analysis of Fourier Transform Infrared Attenuated Total Reflection (FTIR-ATR) and Cyclic Voltammetry (CV), the pH gradient range which is induced by the electrolysis of water is pointed out. Near the positive electrode, OH^- ions moved to the electrode and H⁺ ions produced by electrolysis of water form a pH gradient range which resulting in the closer to electrode, the lower the pH value. Hence, the higher pH value induces the dissolution of hydrogen bonded film. Of course, for the negative electrode, the pH gradient range is the opposite. Then, comparing the film disintegrated near the positive and negative electrode, the pH gradient range makes a big contribution to the dissolution process and results in different morphology and thickness of multilayers. For the hydrogen bonded film on the positive electrode, it takes more time to electrochemically dissolve than the film on the negative electrode. However, the transparency of film on positive electrode is higher than that on negative electrode. Here, the transparency is related to the different ways of film dissolution rather than the disassembled speed. Two different pH gradient ranges induce different disassembled ways that on the positive electrode the film dissolves from outside to inside, and on the negative electrode the film dissolves from inside to outside. The former film always keeps a transparent and uniform topography and the latter film exhibits a cloud and microporous surface. In addition, the pH value affects the stability of hydrogen bond and property of pH gradient range. Results show that the lower pH value is beneficial for the dissolution of film, because the lower pH value can lead the ionic strength in solution to be higher and higher ionic strength allows the electrical conductivity of the solution to be better. Rapid electrolysis of water is able to generate more H⁺ ions on the surface of positive electrode and move more OH^- ions towards the electrode. These two activities let the pH gradient range be able to achieve much stronger acidic and alkaline, which promotes the film dissolution. The applied electrical potential influences the current in the electrolytic bath:higher electrical potential corresponds to higher current and leads to faster dissociation speed. Besides, the proper condition for film dissolution is electrolyte pH=4, applied potential 4 V, and the current about 25 μA. Under this condition, the disassembled speed of film is relatively faster, and the film does not appear cracks but a stable and sustained dissolution.