A quick and high-through LC-MS/MS method has been developed and applied to simultaneously quantify lamivudine (3TC), stavudine(d4T), zidovudine(AZT), efavirenz(EFV), nevirapine(NVP), and Lopinavir/Ritonavir (LPV/RTV) plasma concentrations. A combination of protein precipitation and liquid–liquid extraction was used to extract all compounds. The method showed a good linearity in a concentration range of 20–3200μg/L for 3TC, d4T, AZT, 40-6400μg/L for EFV and NVP, 62.5-10000μg/L for LPV and 12.5-2000μg/L for RTV. Mean intra- and inter-day precision were within ±20% at the LLOQ and ±15% at the other QC level. The accuracy was between 85% and 115% for all seven analytes.