 Temocillin total and free concentrations were determined with high performance liquid chromotographyHPLC with a LiChrospher 100 RP-18 5 μm column Merck AG, using an elution buffer 100 mM Na acetate buffer pH 7acetonitrile 95:5, vv, a flow rate 1 mLmin and a Waters 2690 Alliance System Waters Corp., Milford, MA, USA, with quantification at 235 nm as previously described. 