The invention relates to an autoanalysis method of iron content in water samples. In the method, operated at low pressure, an analytical instrument including a sample flow path, a coloration liquid flow path, an oxidation liquid flow path, a promotion liquid flow path, a low pressure pump, an injection valve, an injection loop, an analysis flow path, an optical flow cell, an optical detector and a computer processing system is used. The method includes the steps of: starting the analytical instrument, mapping a sample spectrum, replacing samples S1 by a series of samples S2 with known concentrations of ferrous ions or ferric ions, repeating the mapping step of the sample spectrum to obtain a series of standard spectra, drawing a working curve by taking the concentrations of standard samples as an abscissa and taking peak heights of the standard sample spectra as a vertical coordinate, comparing the drawn test sample spectra with the standard sample spectra, and calculating the iron content of the sample by a regression equation of the working curve. A coloration liquid R is a neutral red-sulfuric acid aqueous solution, an oxidation liquid O is a potassium periodate aqueous solution, and a promotion liquid C is deionized water.