Development of HPLC assay method for vitamin E derivatives / Nur Azeila Laila Abd. Rahim

Reverse phase high performance liquid chromatography (HPLC) method using UV detection was developed for the analysis of vitamin E derivatives namely a- and y­ tocotrienol. Zorbax 300SB C-18 (5 urn, 250 x 4.6 mm internal diameter) column was used as a stationary phase. Analytes were monitored by UV detection at 295 nm using 0.5:99.5% water in methanol elution. Analysis was run at a flow rate of 1.5 ml/min. The retention time for a-tocotrienol was slightly longer than y-tocotrienol. Calibration curves for a- and y-tocotrienol were linear over concentration range of 10-100 µg/mL with correlation coefficients 0.999 and 0.997, respectively. the coefficient of variation for within-day analysis of a- and y-tocotrienol were all less than 1 %. The recovery of a-tocotrienols and y-tocotrienol from sodium carboxymethylcellulose (CMC) was in the range of 86.691 to 94.582% and from 91.330 to 100.714% respectively. Assay method for tocotrienols using UV detector was successfully developed with reduced retention time in comparison to previous research findings.