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Objective:To establish a method for simultaneously determination of drug loading and entrapment efficiency of total lactones from Andrographis Paniculata loaded solid lipid nanoparticles (TLA-SLN),and to optimize the formula of TLA-SLN.Methods:The free drugs in TLA-SLN were separated by ultrafiltration along with HPLC assay for AND、NEO、DEO and DEH.The separation was performed on a Kromasil C18 column (150 mm × 4.6 mm,5μm) with mixture of acetonitrile-water (30 ∶ 70) as the mobile phase and with column temperature at 35 ℃.