A new method has been developed for separation and determination of zinc,copper,cobalt and nickel based on their colour chelates with 6,6″-dimethyl-4′-phenyl-2,2′:6′,2″-terpyridine(TPY) by ion pair high performance liquid chromatography in a reversed phase mode.The suitable conditions are the methanol-water(80:20,V/V) containing 10mmol/L NaH2PO4-H3PO4 buffer solution(pH=5.0) and 3.0mmol/L sodium dodecyl sulfonate as mobile phase,on a Symmetry C18 column(3.9mmID×150mm),the flow rate of the mobile phase is 1.0mL/min.Spectrophotometric detection was made at 340nm.The detective limits are 0.05,0.10,2.00 and 0.50ng/mL respectively.The method has good selectivity and sensitivity.It has been applied to the separation and determination of zinc,copper,cobalt and nickel in human hair and tobacco and the result is satisfied.