The work aims to study the tribological properties, physicochemical properties and toxicity of in situ preparation of choline amino acid ionic liquid water lubricated additives. Seven kinds of amino acid ionic liquid lubricating additives ([Ch][AA] ILs) were prepared in situ from amino acid and choline in deionized water. Firstly, its tribological properties and physicochemical properties were systematically studied with deionized water as a reference sample. Then, the traditional ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate (L-B104) was used as control sample, and green algae and sea shrimp were used as experimental objects to evaluate the aquatic toxicity of the additives. The kinematic viscosity of aqueous solution increased significantly and showed an increasing trend after adding different kinds of choline amino acid ionic liquid additives. The corrosion of iron bars immersed in aqueous solution containing 400 mmol/L amino acid choline ionic liquid was very slight. When used as lubricant, water-based lubricants with ionic liquid concentration of 400 mmol/L were selected to test the antifriction and antiwear properties of amino acid ionic liquids prepared in situ, and their friction coefficients were smaller than those of water. Toxicity tests showed that the values of EC50 and LC50 of [Ch][AA] ILs were much higher than those of L-B104. Compared with deionized water, choline amino acid ionic liquid as water lubricating additive exhibits excellent lubrication performance on steel/steel friction pairs. Among them, [Ch][Trp] containing aromatic heterocycles has the best antifriction and antiwear properties, and its tribological coefficient and wear volume are the smallest. This may be attributed to the significant increase in the viscosity of aqueous solution containing the ionic liquid additive due to the introduction of aromatic heterocycles. A strong physical adsorption protective film is formed during the friction process, which makes the amino acid choline ionic liquid possess superior wear resistance. In the toxicity test, the toxicity of amino acid choline ionic liquid lubricant is much less than that of traditional ionic liquid (L-B104). It has no toxic effect on the experimental organisms.