The metallothionein-like(MT-like) was separated and purified by using a high-pressure gel column,then a new hyphenated technique of high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry(HPLC-ICP-MS) was established based on the protein ultraviolet absorption and the metal element combined,and with Tris-acetate buffer solution(20 mmol/L,pH 7.5) as mobile phase at a flow rate of 0.5 mL/min.The separation effect and reproducibility of bovine serum protein from Rabbit Liver Zn-Metallothionein(Zn-MTs) fraction were studied,and the linear relationship,detection limit and correlation of UV and ICP-MS were also investigated.The experimental results indicated that it had a good reproducibility on Zn-MTs group separation with RSD(n=6) less than 5%,and Zn signal had a good quantity relationship with UV absorption in Zn-MTs with a linear correlation coefficient of 0.999 5.The detection limit of Zn-MTs protein was relatively low that was based on Zn detected by ICP-MS.