The Benefits of Ultra-High-Performance Supercritical Fluid Chromatography in Determination of Lipophilic Vitamins in Dietary Supplements
- 16 Downloads
Nine tocopherol derivatives including four tocopherols, four tocotrienols, and one esterified form of α-tocopherol acetate were analyzed using ultra-high-performance supercritical fluid chromatography (UHPSFC) with diode array detection. The separation was carried out on BEH 2-EP column using shallow gradient elution of 1–4% methanol and CO2 in 5 min. Complementary ultra-high-performance liquid chromatography method was developed using Ascentis Express F5 core–shell stationary phase and isocratic elution with 75% acetonitrile/methanol (1:1) and 25% water. This separation took 12 min that is the shortest liquid chromatography run in analysis of all tocopherol derivatives reported so far. However, our UHPSFC method resulted in more than two times faster analysis, and allowed direct injection of heptane solutions and extracts from liquid–liquid extraction. These features made UHPSFC a method of choice for subsequent analysis of dietary supplements containing different forms of vitamin E. Sample preparation methods were optimized for individual dosage forms of drops, capsules, tablets, and granulate.
KeywordsUltra-high-performance supercritical fluid chromatography Ultra-high-performance liquid chromatography Liquid–liquid extraction Vitamin E Dietary supplements Method validation
The authors gratefully acknowledge the Grant project SVV no. 260412/2018 and the project EFSA-CDN (no. CZ.02.1.01/0.0/0.0/16_019/0000841) co-funded by ERDF.
Kateřina Plachká acknowledges funding of the Grant project SVV no. 260412/2017. Lucie Nováková and František Švec acknowledge the funding of the project EFSA-CDN (no. CZ.02.1.01/0.0/0.0/16_019/0000841) co-funded by ERDF.
Compliance with Ethical Standards
Conflict of interest
All authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- 2.San Andrés MP, Otero J, Vera S (2011) High performance liquid chromatography method for the simultaneous determination of α-, γ- and δ-tocopherol in vegetable oils in presence of hexadecyltrimethylammonium bromide/n-propanol in mobile phase. Food Chem 126(3):1470–1474. https://doi.org/10.1016/j.foodchem.2010.11.161 CrossRefGoogle Scholar
- 5.Oberson JM, Campos-Gimenez E, Riviere J, Martin F (2018) Application of supercritical fluid chromatography coupled to mass spectrometry to the determination of fat-soluble vitamins in selected food products. J Chromatogr B Anal Technol Biomed Life Sci 1086:118–129. https://doi.org/10.1016/j.jchromb.2018.04.017 CrossRefGoogle Scholar
- 9.Sieniawska E, Baj T, Sawicki R, Wanat A, Wojtanowski KK, Ginalska G, Zgorka G, Szymanska J (2017) LC–QTOF–MS analysis and activity profiles of popular antioxidant dietary supplements in terms of quality control. Oxid Med Cell Longev 2017:8692516. https://doi.org/10.1155/2017/8692516 CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Young AL, Woodlee JW, McGuffin MM (2018) Chap. 9—dietary supplements. In: Pacifici E, Bain S (eds) An overview of FDA regulated products. Academic Press, New York, pp 199–215. https://doi.org/10.1016/B978-0-12-811155-0.00009-0 CrossRefGoogle Scholar
- 15.Chase GW Jr, Eitenmiller RR, Long AR (1999) A liquid chromatographic method for analysis of all-rac-alpha-tocopheryl acetate and retinyl palmitate in medical food using matrix solid-phase dispersion in conjunction with a zero reference material as a method development tool. J AOAC Int 82(1):107–111PubMedGoogle Scholar
- 21.Zhao J, Yang G, Duan H, Li J (2001) Determination of synthesized alpha-vitamin E by micellar electrokinetic chromatography. Electrophoresis 22 (1):151–154. https://doi.org/10.1002/1522-2683(200101)22:1%3C151::Aid-elps151%3E3.0.Co;2-0 CrossRefPubMedGoogle Scholar
- 22.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. https://doi.org/10.1016/j.chroma.2012.04.042 CrossRefPubMedGoogle Scholar
- 23.Pilařová V, Gottvald T, Svoboda P, Novák O, Benešová K, Běláková S, Nováková L (2016) Development and optimization of ultra-high performance supercritical fluid chromatography mass spectrometry method for high-throughput determination of tocopherols and tocotrienols in human serum. Anal Chim Acta 934:252–265. https://doi.org/10.1016/j.aca.2016.06.008 CrossRefPubMedGoogle Scholar
- 27.Petruzziello F, Grand-Guillaume Perrenoud A, Thorimbert A, Fogwill M, Rezzi S (2017) Quantitative profiling of endogenous fat-soluble vitamins and carotenoids in human plasma using an improved UHPSFC–ESI-MS interface. Anal Chem 89(14):7615–7622. https://doi.org/10.1021/acs.analchem.7b01476 CrossRefPubMedGoogle Scholar
- 28.ICH-Expert-Working-Group (1994) ICH harmonised tripartite guideline: validation of analytical procedures: text and methodology. https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf. Accessed Aug 2018