Direct high-performance liquid chromatographic analysis of d-tocopheryl acid succinate and derivatives

A method of analysis of a Vitamin E derivative d-tocopheryl acid succinate (TS) in biological fluids and commercially available products is necessary to study the kinetics of in vitro and in vivo metabolism, tissue distribution, and content uniformity. A simple and inexpensive high-performance liquid chromatographic method was developed for the direct determination of d-tocopheryl acid succinate in commercially available products, rat serum, and rat tissues. This method can also be applied to the determination of 15 Vitamin E derivatives. Rat serum (0.1 ml) was extracted with sodium dodecyl sulfate, ethanol, hexane, and then dried under nitrogen gas after addition of the internal standard, dl-α-tocopherol acetate. Separation was achieved on a C18 column with UV detection at 205 nm. The calibration curve for d-tocopheryl acid succinate was linear ranging from 0.025 to 100 μg/ml. The mean extraction efficiency was >92%. Precision of the assay was <5% (CV), and was within 5% at the limit of quantitation (0.025 μg/ml). Bias of the assay was lower than 5%, and was within 5% at the limit of quantitation. The assay was applied successfully to the serum and tissue distribution of d-tocopheryl acid succinate in rats, various Vitamin E derivatives, and content uniformity in commercially available products containing d-tocopheryl acid succinate.