A method utilizing both MALDI-TOF and HPLC-UV analyses was developed for the determination of different human insulin injection products. Human, bovine and porcine insulins as well as other analogue insulins could be identified by the MALDI-TOF analysis based on the results on the accurate intact molecular weights and the high sequence coverage on the detection of insulin B chain peptides. Assay of the identified insulin was accomplished by a reversed-phase HPLC analysis employing a C5 column, UV detection at 215 nm and gradient elution with a mobile phase composed of acetonitrile and 0.1% of aqueous trifluoroacetic acid solution. As this method enables fast and accurate screening of insulins, it is considered suitable for high throughput analyses of human insulin injection products being sold in Hong Kong with an aim to safeguard the public health in relation to its use.