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Determination of Tocopherols and Tocotrienols in Palm Oils

Explore the significance of tocols (vitamin E) in palm oil, their antioxidant properties, and analytical methods for determination using high-performance liquid chromatography. Learn about the chemical structures and detection techniques involved.

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Determination of Tocopherols and Tocotrienols in Palm Oils

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  1. Determination Of Tocopherols And Tocotrienols In Palm Oils. Name : Ifianly Bin Karim 156496 Supervisor : Dr. Mohd. NazrimMarikar

  2. What is actually palm oil ? • Palm oil is natural oil extracted from the fleshy orange-red mesocarp of the fruit fromthe oil palm (Elaeisguineensis).

  3. Figure 1 – General flow of production of Palm Oil.

  4. What is tocopherols and tocotrienols ? Both are known as tocols (vitamin E)

  5. Amphipathicand lipid-soluble compounds • a polar chromanol ring  • hydrophobic 16-carbon side chain • Each consist of 4 isomers -α-, -β-, -ϒ-, -δ- • palm oil – only 5 (α-T) , (α-T₃) , (ϒ-T) , (ϒ-T₃) , (δ-T₃) T = tocopherol T₃ = tocotrienol

  6. Figure 2- Chemical structures of tocopherols and tocotrienols.

  7. Importance of Tocols • Chemical property – antioxidant activity • The ability to protect polyunsaturated lipids from oxidation. • The palm’s tocol isomers areknown for their antioxidant properties which make them popular ingredients in pharmaceuticals, cosmetics formulations and health supplements.

  8. How to determine tocols ?

  9. Tocols can be analysed by gas chromatography, both with flame-ionization and mass spectrometric detection, but the methods that are usually recommended involve high-performance liquid chromatography with fluorescence detection. Gas chromatography High performance liquid chromatography

  10. High performances liquid chromatography • both normal-phase (NP) and reversed-phase (RP) chromatography. • All eight tocols have been base-line separated by NP-HPLC using silica columns. • Using 4–5 % (v/v) of 1,4-dioxane in hexane as the mobile phase with several silica columns, tocols were always eluted in the following order:  α-tocopherol< α-tocotrienol < β-tocopherol < γ-tocopherol< β-tocotrienol < γ-tocotrienol < δ-tocopherol < δ-tocotrieno

  11. Continued… • RP-HPLC has the advantage over using NP-HPLC that less hazardous mobile phases e.g. methanol or ethanol. • Saponification - removal of acyl lipids to avoid contamination of the RP column. • By using a C30-bonded silica column and methanol as the mobile phase, it was possible to separate α-tocopherol and all tocotrienolsin palm oil.

  12. Detection of tocols • Tocols absorb UV light at λ=290-300 nm • Fluorescence detection (FLD) • Better sensitivity and higher selectivity of HPLC analysis • Excitation wavelength of 290-296 nm and an emission wavelength of 325-330 nm are commonly used

  13. Continued… • Mass spectrometry (MS) has also been used to detect tocols. • Negative ion APCI (atmospheric pressure chemical ionization) was found the best choice among negative and positive ion electrospray and APCI ionization modes to detect and quantify tocopherols.

  14. References • Kamal-Eldin, A. and Appelqvist, L. The chemistry and antioxidant properties of tocopherols and tocotrienols. Lipids, 31, 671-701 (1996). • Eitenmiller, R.R. and Lee, J. Vitamin E, Food Chemistry, Composition, and Analysis. Pages 540. Marcel Dekker Inc., New York, NY (2004). • Kamal-Eldin, A., Gorgen, S., Pettersson, J. and Lampi, A.M. Normal-phase high-performance liquid chromatography of tocopherols and tocotrienols - comparison of different chromatographic columns. J. Chromatogr. A, 881, 217-227 (2000).

  15. Bustamante-Rangel, M., Delgado- Zamarreno, M.M., Sanchez-Perez, A. and Carabias-Martinez, R.. Determination of tocopherols and tocotrienols in cereals by pressurized liquid extraction-liquid chromatography-mass spectrometry. Anal. Chim. Acta, 587, 216-221 (2007). • Ng, M.H., Choo, Y.M., Ma, A.N., Chuah, C.H. and Hashim, M.A. Separation of vitamin E (tocopherol, tocotrienol, and tocomonoenol) in palm oil. Lipids, 39, 1031-1035 (2004). • Abidi, S.L. Tocol-derived minor constituents in selected plant seed oils. J. Am. Oil Chem. Soc., 80, 327-333 (2003).

  16. Summary • Palm oil ( Elaeisguineensis) • Tocols consist of tocopherols and tocotrienols • Important of tocols as in the antioxidant activity. • Highly recommended to determine tocols by using high performance liquid chromatography and fluorescence detection.

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