Skip to main content
SpringerLink
Log in

Rapid Reversed-Phase Chromatographic Method for Determination of Eight Vitamin E Isomers and γ-Oryzanols in Rice Bran and Rice Bran Oil

  • Published:
Food Analytical Methods Aims and scope Submit manuscript

Abstract

Rice bran and rice bran oil contain high amounts of lipid soluble phytochemicals such as tocopherols, tocotrienols and γ-oryzanols with potential health benefits. A reversed-phase high-performance liquid chromatographic method for the quantitation of these phytochemicals was developed. These phytochemicals were extracted using rapid one-step methanol extraction and separated on a Kinetex pentafluorophenyl phase column for quantitation using fluorescence and ultraviolet detectors. Separation was carried out within 20 min at a flow rate of 1.0 mL/min using a gradient elution of mobile phase consisting of methanol and water at column temperature of 30 °C. A good separation of all eight tocol isomers was achieved while the γ-oryzanols were separated near to baseline. Extraction yields of tocopherols, tocotrienols and γ-oryzanols from saponification medium with different pH conditions were studied. Total γ-oryzanols extracted from alkaline medium were significantly lower than those from the neutral and acidic mediums (p < 0.05), while total tocols extracted from these mediums were not significantly different. The method gave a good calibration linear response (R 2 > 0.9994), low detection limit (3–34 ng/mL) and high precision (RSD < 7.5 %) with satisfactory recovery (R% = 81–116 %). The advantages of this method over available reverse-phase methods are the better resolution of β and γ tocol isomers with the γ-oryzanols, usage of non-halogenated mobile phase and shorter analysis time, and it is compatible with the standard laboratorial chromatographic equipment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Bergman C, Xu Z (2003) Genotype and environment effects on tocopherol, tocotrienol, and γ-oryzanol contents of Southern US rice. Cereal Chem 80:446–449

    Article  CAS  Google Scholar 

  • Chen MH, Bergman C (2005) A rapid procedure for analysing rice bran tocopherol, tocotrienol and γ-oryzanol contents. J Food Compos Anal 18:319–331

    Article  Google Scholar 

  • D’Ambrosio M (2013) Performances of CN-columns for the analysis of γ-oryzanol and its p-coumarate and caffeate derivatives by normal phase HPLC and a validated method of quantitation. Food Chem 138:2079–2088

    Article  Google Scholar 

  • Das PK, Chaudhuri A, Kaimal TNB, Bhalerao UT (1998) Isolation of γ-oryzanol through calcium ion induced precipitation of anionic micellar aggregates. J Agric Food Chem 46:3073–3080

    Article  CAS  Google Scholar 

  • Diack M, Saska M (1994) Separation of vitamin E and γ-oryzanols from rice bran by normal-phase chromatography. J Am Oil Chem Soc 71:1211–1217

    Article  CAS  Google Scholar 

  • Eitenmiller RR, Ye L, Landen W (2008) Vitamin analysis for the health and food sciences. CRC Press, Boca Raton

    Google Scholar 

  • Evershed RP, Spooner N, Prescott MC, John Goad L (1988) Isolation and characterisation of intact steryl ferulates from seeds. J Chromatogr A 440:23–35

    Article  CAS  Google Scholar 

  • Grebenstein N, Frank J (2012) Rapid baseline-separation of all eight tocopherols and tocotrienols by reversed-phase liquid-chromatography with a solid-core pentafluorophenyl column and their sensitive quantification in plasma and liver. J Chromatogr A 1243:39–46

    Article  CAS  Google Scholar 

  • Hakala P, Lampi A-M, Ollilainen V, Werner U, Murkovic M, Wähälä K, Karkola S, Piironen V (2002) Steryl phenolic acid esters in cereals and their milling fractions. J Agric Food Chem 50:5300–5307

    Article  CAS  Google Scholar 

  • Heinemann RJB, Xu Z, Godber JS, Lanfer-Marquez UM (2008) Tocopherols, tocotrienols, and γ-oryzanol contents in japonica and indica subspecies of rice (Oryza sativa L.) cultivated in Brazil. Cereal Chem 85:243–247

    Article  CAS  Google Scholar 

  • Huang S, Ng L (2011) An improved high-performance liquid chromatographic method for simultaneous determination of tocopherols, tocotrienols and γ-oryzanol in rice. J Chromatogr A 1218:4709–4713

    Article  CAS  Google Scholar 

  • Inoue T, Tatemori S, Muranaka N, Hirahara Y, Homma S, Nakane T, Takano A, Nomi Y, Otsuka Y (2012) The identification of vitamin E homologues in medicinal plant samples using ESI (+)-LC-MS3. J Agric Food Chem 60:9581–9588

    Article  CAS  Google Scholar 

  • Lerma-Garcia M, Herrero-Martinez J, Simo-Alfonso E, Mendonca CR, Ramis-Ramos G (2009) Composition, industrial processing and applications of rice bran γ-oryzanol. Food Chem 115:389–404

    Article  CAS  Google Scholar 

  • Panfili G, Fratianni A, Irano M (2003) Normal phase high-performance liquid chromatography method for the determination of tocopherols and tocotrienols in cereals. J Agric Food Chem 51:3940–3944

    Article  CAS  Google Scholar 

  • Richheimer SL, Kent MC, Bernart MW (1994) Reversed-phase high-performance liquid chromatographic method using a pentafluorophenyl bonded phase for analysis of tocopherols. J Chromatogr A 677:75–80

    Article  CAS  Google Scholar 

  • Rogers E, Rice S, Nicolosi R, Carpenter D, McClelland C, Romanczyk L Jr (1993) Identification and quantitation of γ-oryzanol components and simultaneous assessment of tocols in rice bran oil. J Am Oil Chem Soc 70:301–307

    Article  CAS  Google Scholar 

  • Seitz LM (1989) Stanol and sterol esters of ferulic and p-coumaric acids in wheat, corn, rye, and triticale. J Agric Food Chem 37:662–667

    Article  CAS  Google Scholar 

  • Shammugasamy B, Ramakrishnan Y, Ghazali HM, Muhammad K (2013) Combination of saponification and dispersive liquid–liquid microextraction for the determination of tocopherols and tocotrienols in cereals by reversed-phase high-performance liquid chromatography. J Chromatogr A 1300:31–37

    Article  CAS  Google Scholar 

  • Stoggl W, Huck C, Wongyai S, Scherz H, Bonn G (2005) Simultaneous determination of carotenoids, tocopherols, and γ-oryzanol in crude rice bran oil by liquid chromatography coupled to diode array and mass spectrometric detection employing silica C30 stationary phases. J Sep Sci 28:1712–1718

    Article  Google Scholar 

  • Vinas P, Bravo-Bravo M, Lopez-García I, Pastor-Belda M, Hernandez-Cordoba M (2014a) Pressurized liquid extraction and dispersive liquid-liquid microextraction for determination of tocopherols and tocotrienols in plant foods by liquid chromatography with fluorescence and atmospheric pressure chemical ionization-mass spectrometry detection. Talanta 119:98–104

    Article  CAS  Google Scholar 

  • Vinas P, Pastor-Belda M, Campillo N, Bravo-Bravo M, Hernandez-Cordoba M (2014b) Capillary liquid chromatography combined with pressurized liquid extraction and dispersive liquid-liquid microextraction for the determination of vitamin E in cosmetic products. J Pharm Biomed Anal 94:173–179

    Article  CAS  Google Scholar 

  • Xu Z, Godber JS (1999) Purification and identification of components of γ-oryzanol in rice bran oil. J Agric Food Chem 47:2724–2728

    Article  CAS  Google Scholar 

  • Xu Z, Godber JS (2000) Comparison of supercritical fluid and solvent extraction methods in extracting γ-oryzanol from rice bran. J Am Oil Chem Soc 77:547–551

    Article  CAS  Google Scholar 

  • Yu S, Nehus ZT, Badger TM, Fang N (2007) Quantification of vitamin E and γ-oryzanol components in rice germ and bran. J Agric Food Chem 55:7308–7313

    Article  CAS  Google Scholar 

Download references

Conflict of interest

Balakrishnan Shammugasamy declares that he has no conflict of interest. Kharidah Muhammad declares that she has no conflict of interest. Yogeshini Ramakrishna declares that she has no conflict of interest. Farhiah Manan declares that she has no conflict of interest. This article does not contain any studies with human or animal subjects.

Author information

Authors and Affiliations

  1. UPM-BERNAS Research Laboratory, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Darul Ehsan, Malaysia

    Balakrishnan Shammugasamy, Yogeshini Ramakrishnan, Farhiah Manan & Kharidah Muhammad

Authors
  1. Balakrishnan Shammugasamy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yogeshini Ramakrishnan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Farhiah Manan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kharidah Muhammad
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kharidah Muhammad.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shammugasamy, B., Ramakrishnan, Y., Manan, F. et al. Rapid Reversed-Phase Chromatographic Method for Determination of Eight Vitamin E Isomers and γ-Oryzanols in Rice Bran and Rice Bran Oil. Food Anal. Methods 8, 649–655 (2015). https://doi.org/10.1007/s12161-014-9929-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12161-014-9929-0

Keywords

Search

Navigation