Tocopherols are essential antioxidant vitamins primarily found in the chloroplasts of plants.
Tocopherols’ extreme lipophilicity can challenge the isolation of the different vitamers. This is especially common in reversed-phase chromatography, where water is omitted from the eluent section.
It is also important to note that the different vitamers of tocopherols (α, ß, γ, δ) differ according to methylations on the fused phenoltetrahydropyran core.
Research has shown that tocopherols regulate cellular signaling, proper gene expression, cell proliferation, and serum cholesterol reduction.1,2
It has also been posited that tocopherols protect polyunsaturated lipids from oxidation. Tocopherols are typically regarded as lipid peroxyl radical scavengers able to react with both reactive oxygen and reactive nitrogen species that can lead to disease if left unchecked.3
Tocopherols can be found in various plant seed oils, grains, dairy products, and meats.4
In the example presented here, the vitamers were purified using the distinct adsorption profile linked to Hamilton Company’s PRP-1 HPLC column. This column’s related pore structure and lipophilicity effectively isolated three of the vitamers while continuing to achieve baseline resolution and good peak shape.
This separation centered around the use of tetrahydrofuran with acetonitrile. Using THF as an eluent was vital in ensuring improved mass transfer kinetics in polymeric stationary phases like PRP-1. This is particularly important when there is a high degree of lipophilic interaction with the solute.
The PS-DVB core ensures excellent reproducibility and unparalleled longevity, simplifying and streamlining the analysis process.
Chromatogram and compound results
Source: Hamilton Company
| Column Information |
| Packing Material |
PRP-1, 5 µm |
| Dimensions |
150 x 4.1 mm |
| P/N |
79444 |
| Chromatographic Conditions |
| Gradient |
0.0–10.00 min. 80–100 % B
10.01–12.00 min 100 % B |
| Temperature |
Ambient |
| Injection Volume |
5 µL |
| Detection |
UV at 292 nm |
| Eluent A |
20 mM NH4 OAc |
| Eluent B |
Acetonitrile/Tetrahydrofuran (1:1) |
| Flow Rate |
1.0 mL/min |
Compounds:
- Δ-tocopherol
- ß+γ-tocopherol
- α-tocopherol
Image Credit: Hamilton Company
References and further reading
- Traber, M.G. and Atkinson, J. Free Rad. Biol. Med., 43, 4-15 (2007).
- Sen, C.K., Khanna, S. and Roy, S. Mol. Aspects Med., 28, 692-728 (2007).
- Niki, E. Free Rad. Biol. Med., 49, 503-515 (2010).
- Ruperez, F. J., Martin, D., Herrera, E. and Barbas, C. J. Chromatogr. A, 935, 45-69 (2001).
Acknowledgments
Produced from materials originally authored by Adam L. Moore PhD. from Hamilton Company.
About Hamilton Company
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Hamilton Company specializes in the development, manufacturing and customization of precision measurement devices, automated liquid handling workstations, and sample management systems. Hamilton's processes are optimized for quality and flexibility. Whether it's a custom needle with a quick delivery time frame, a special length pH sensor, or a comprehensive solution to fully automate your assay workflow, trust that Hamilton products will always meet your needs.
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