Abstract: OBJECTIVE To prepare monoester and diester from D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) using a preparative HPLC method, and to establish determination methods of monoester, diester, vitamin E succinate, α-tocopheryl and polyethylene glycol 1000 in TPGS. METHODS A preparative HPLC method with a DAC-HB50 C₁₈dynamic axis compress chromatographic column (50 mm×250 mm, 10 μm) was developed using the mobile phase of water (A)-ethanol (B) in a gradient mode at a flow rate of 75 mL·min^-1. Chromatograms were observed by a UV detector with wavelength set to 285 nm. Collected target product solution was evaporated by rotary evaporation. The target products were identified. An HPLC method with a Waters XBridge C₈ column (4.6 mm×250 mm, 5 μm) for the determination of prepared components was developed using a gradient elution of 0.1% formic acid solution and acetonitrile-isopropanol (1:1) at a flow rate of 1.0 mL·min^-1. Chromatograms were observed by a UV detector with wavelength set to 285 nm. The HPLC method was used for determination of monoester, diester, vitamin E succinate and α-tocopheryl in TPGS. An HPLC-ESLD method for the determination of polyethylene glycol 1000 in TPGS was developed with the same column and mobile phase but different gradient elution of HPLC method. Chromatograms were observed by an ELSD detector with drift tube temperature of 90.4℃ and gas flow rate of 2.3 L·min^-1. RESULTS The purity of prepared monoester and diester were 99.1% and 99.8%. The methods were validated and they were found that good resolution, linearity (r ≥ 0.999 5) within the certain range of five components and average recoveries of 98.2%-99.7% were achieved. CONCLUSION The preparative HPLC method is used successfully for the preparation of monoester and diester. Two validated detective HPLC methods can be used for detection of monoester, diester, vitamin E succinate, α-tocopheryl and polyethylene glycol 1000 in TPGS.