Quantitative Determination of α-Tocopherol and α-Tocopheryl Acetate in Pharmaceutical and Supplementary Soft Capsules by High Performance Liquid Chromatography

Year 2025, Issue: 26, 500 - 511, 31.08.2025

Nükte Topraksever , Gamze Özgül Artuç

https://doi.org/10.38079/igusabder.1589685

Abstract

Aim: This study aims to determine the amounts of α-tocopherol (α-toc) and α-tocopheryl acetate (α-tocAc) in soft pharmaceutical capsules and supplements using a chromatographic method. Additionally, method validation parameters for both analytes were evaluated.
Method: Analyses were performed using a chromatographic method. As part of the method validation, recovery (% recovery), repeatability (% RSD), linear regression coefficient (R²), limit of detection (LOD), and limit of quantification (LOQ) values were determined.
Results: The recovery values were found to be above 95%. Repeatability (%RSD) was calculated as <3%. The linearity (R²) values were obtained as 0.999. For α-toc: LOD: 0.131 µg/mL and 2.228 µg/mL, LOQ: 0.396 µg/mL and 6.752 µg/mL. For α-tocAc: LOD: 0.260 µg/mL, LOQ: 0.787 µg/mL.
Conclusion: The chromatographic method used in this study enabled the determination of α-toc and α-tocAc with high accuracy and repeatability. This method can be considered a reliable alternative for the analysis of these compounds in pharmaceutical products.

Keywords

α-tocopherol , α-tocopheryl acetate , HPLC , soft capsule

Farmasötik ve Takviye Edici Yumuşak Kapsüllerdeki α-Tokoferol ve α-Tokoferil Asetat’ın Yüksek Performanslı Sıvı Kromatografisi ile Kantitatif Tayini

Abstract

Amaç: Bu çalışmada, yumuşak farmasötik kapsüller ve takviyelerde bulunan α-tokoferol (α-tok) ve α-tokoferil asetat (α-tokAs) miktarlarının kromatografik yöntem kullanılarak belirlenmesi amaçlanmıştır. Ayrıca, her iki analit için yöntem doğrulama (validasyon) parametreleri değerlendirilmiştir.
Yöntem: Analizler kromatografik bir yöntemle gerçekleştirilmiştir. Yöntem validasyonu kapsamında geri kazanım, tekrarlanabilirlik, doğrusal regresyon katsayısı, LOD ve LOQ değerleri belirlenmiştir.
Bulgular: Geri kazanım değerleri %95’in üzerinde bulunmuştur. Tekrarlanabilirlik (%RSD) <%3 olarak hesaplanmıştır. Doğrusallık (R²) değerleri 0.999 olarak elde edilmiştir. α-tok için: LOD: 0.131 µg/mL ve 2.228 µg/mL, LOQ: 0.396 µg/mL ve 6.752 µg/mL α-tokAs için: LOD: 0.260 µg/mL, LOQ: 0.787 µg/mL.
Sonuç: Çalışmada kullanılan kromatografik yöntem, α-tok ve α-tokAs miktarlarının yüksek doğruluk ve tekrarlanabilirlik ile belirlenmesini sağlamıştır. Yöntem, farmasötik ürünlerdeki bu bileşiklerin analizi için güvenilir bir alternatif olarak değerlendirilebilir.

Keywords

α-tokoferol , α-tokoferil asetat , HPLC , yumuşak kapsül

References

• 1. Asgher M, Waseem A, Yaqoob M, Nabi A. Flow injection chemiluminescence determination of retinol and α-tocopherol in blood serum and pharmaceuticals. Anal Lett. 2011;44(1-3):12-24.
• 2. Traber MG, Packer L. Vitamin E: Beyond antioxidant function. Am J Clin. 1995;62(6):1501-1509.
• 3. Dongala T, Palakurthi AK. Stability-indicating LC method for the simultaneous determination of methyl paraben, propyl paraben, butylated hydroxytoluene and alpha-tocopherol contents in marijuana capsules. J Iran Chem Soc. 2020;17(3):631-638.
• 4. Jiang Q. Natural forms of vitamin E: Metabolism, antioxidant, and anti-inflammatory activities and their role in disease prevention and therapy. Free Radic Biol Med. 2014;72:76-90.
• 5. Maldonado A, Riquelme N, Muñoz-Fariña O, García O, Arancibia C. Stability and bioaccessibility of α-tocopherol-enriched nanoemulsions containing different edible oils as carriers. LWT. 2023;174.
• 6. Gianeti MD, Gaspar LR, De Camargo FB, Campos PMBGM. Benefits of combinations of vitamin A, C and e derivatives in the stability of cosmetic formulations. Molecules. 2012;17(2):2219-2230.
• 7. Mandelli De Almeida M, Rosana C, De Castro Lima R, et al. Stability evaluation of tocopheryl acetate and ascorbyl tetraisopalmitate in isolation and incorporated in cosmetic formulations using thermal analysis. Braz J Pharm Sci. 2010;46:129-134.
• 8. Ben-Shabat S, Kazdan Y, Beit-Yannai E, Sintov AC. Use of alpha-tocopherol esters for topical vitamin e treatment: Evaluation of their skin permeation and metabolism. J Pharm Pharmacol. 2013;65(5):652-658.
• 9. Anastassakis K. Androgenetic Alopecia From A to Z. Springer International Publishing; 2022.
• 10. Retzlaff D, Dörfler J, Kutschan S, Freuding M, Büntzel J, Hübner J. The vitamin E isoform α-tocopherol is not effective as a complementary treatment in cancer treatment: A systematic review. Nutr Cancer. 2022;74(7):2313-2336.
• 11. Wallert M, Ziegler M, Wang X, et al. α-Tocopherol preserves cardiac function by reducing oxidative stress and inflammation in ischemia/reperfusion injury. Redox Biol. 2019;26.
• 12. Lee ARY Bin, Tariq A, Lau G, Tok NWK, Tam WWS, Ho CSH. Vitamin E, Alpha-tocopherol, and its effects on depression and anxiety: A systematic review and meta-analysis. Nutrients. 2022;14(3).
• 13. Emecen G. Effect of α- tocopherol and ascorbic acid on the genotoxicity by gamma-irradiation in drosophila melanogaster. Hacettepe J Biol Chem. 2022;50(1):93-98.
• 14. Waseem A, Rishi L, Yaqoob M, Nabi A. Flow-injection determination of retinol and tocopherol in pharmaceuticals with acidic potassium permanganate chemiluminescence. Anal Sci. 2009;25:407-412.
• 15. U.S. Food & Drug Administration. Select Committee on GRAS Substances (SCOGS) Opinion: Alpha-Tocopherol Acetate, Tocopherols, 2015.
• 16. Loughrill E, Govinden P, Zand N. Vitamins A and e content of commercial infant foods in the UK: A cause for concern? Food Chem. 2016;210:56-62.
• 17. Karpińska J, Mikołuć B, Motkowski R, Piotrowska-Jastrzebska J. HPLC method for simultaneous determination of retinol, α-tocopherol and coenzyme Q10 in human plasma. J Pharm Biomed Anal. 2006;42(2):232-236.
• 18. Jaiswal P V, Ijeri VS, Srivastava AK. Voltammetric behavior of-tocopherol and its determination using surfactant + ethanol + water and surfactant + acetonitrile + water mixed solvent systems. Anal Chim Acta. 2001;441:201-206.
• 19. Özgül Artuç G. Farmasötik yumuşak kapsülde α-tokoferolün spektrofotometri ile kantitatif tayini. Experimed. 2020;10(2):72-76.
• 20. Moreno P, Salvado VS, Salvadó S. Determination of eight water-and fat-soluble vitamins in multi-vitamin pharmaceutical formulations by high-performance liquid chromatography. J Chromatogr A. 2000;870:207-215.
• 21. Ruperez FJ, Martın D, Herrera E, Barbas C, Barbas B. Chromatographic analysis of a-tocopherol and related compounds in various matrices. J Chromatogr A. 2001;935:45-69.
• 22. Bonifacio FN, Giocanti M, Reynier JP, Lacarelle B, Nicolay A. Development and validation of HPLC method for the determination of cyclosporin a and its impurities in Neoral® capsules and its generic versions. J Pharm Biomed Anal. 2009;49(2):540-546.
• 23. Luhua Z, Ying T, Zhengyu Z, Guangji W. Determination of aa-tocopherol in the traditional chinese medicinalpreparation sea buckthorn oil capsule by non-aqueous reversed phase-HPLC. Chem Pharm Bull (Tokyo). 2004;52(1):150-152.
• 24. EMEA. ICH Topic Q 2 (R1) Validation of Analytical Procedures.; 1995. http://www.emea.eu.int.
• 25. Goudarzi N, Farsimadan S, Arab Chamjangali M, Bagherian GA. Development of coupled ultrasound-assisted and reversed-phase dispersive liquid-liquid microextraction before high-performance liquid chromatography for the sensitive determination of vitamin A and vitamin E in oil samples. J Sep Sci. 2015;38(18):3254-3261.
• 26. Pasias IN, Kiriakou IK, Papakonstantinou L, Proestos C. Determination of vitamin E in cereal products and biscuits by GC-FID. Foods. 2018;7(1).
• 27. Kadioglu Y, Demirkaya F, Kursat Demirkaya A. Quantitative determination of underivatized α-tocopherol in cow milk, vitamin and multivitamin drugs by GC-FID. Chromatographia. 2009;70(3-4):665-670.
• 28. Demirkaya-Miloglu F, Kadioglu Y, Senol O, Yaman ME. Spectrofluorimetric determination of a-tocopherol in capsules and human plasma. Indian J Pharm Sci. 2013;5(75):563-568.
• 29. Ashour H, Maher H, Kamel S, El-Yazbi F. Novel, cost-effective, eco-friendly spectrophotometric method for the determination of beta-carotene, vitamin C, and vitamin E in their ternary mixtures: Greenness and whiteness appraisal. J Adv Pharm Sci. 2024;1(1):22-28.
• 30. Méjean M, Brunelle A, Touboul D. Quantification of tocopherols and tocotrienols in soybean oil by supercritical-fluid chromatography coupled to high-resolution mass spectrometry. Anal Bioanal Chem. 2015;407(17):5133-5142.
• 31. Jeelani S, Kassymbek Z. Simultaneous determination of ten fat-soluble vitamins by ultra-performance liquid chromatography in multivitamins with mineral capsules. Journal of Separation Science, 2025; 48:e70167
• 32. Wang LH, Wang JF. Determination of retinoids in human serum, tocopherol and retinyl acetate in pharmaceuticals by RP-LC with electrochemical detection. J Pharm Biomed Anal. 2001;25:785-793.