Determination of Rebaudioside A and Stevioside in Beverage and Stevia Samples by High-Performance Liquid Chromatography
Abstract
The optimized conditions for the determination of two steviol glycosides, namely rebaudioside A and stevioside in beverage and stevia samples by using high-performance liquid chromatography coupled with diode array detector was studied. Analytical column was Luna C18 (4.6 mm x 250 mm, 5 micrometer). Both analytes were detected at 210 nm. It was found that the optimized mobile phase comprised acetonitrile and water (pH 3.5) in ratio of 35:65 (%v/v). The flow rate of mobile phase was 0.60 mL/min. Under these optimized conditions, analysis time was about 8 min. The limits of detection were 0.03 mg/L for rebaudioside A and stevioside, and the limits of quantitation were 0.10 mg/L for rebaudioside A and stevioside. Correlation coefficients of calibration curves from 0.01 to 5.00 mg/L were 0.9990 for rebaudioside A, and 0.9998 for stevioside. Intra-day precisions in terms of percent relative standard deviation of retention times and peak areas were 0.17-2.36 for rebaudioside A, and 00.10-2.04 for stevioside. Inter-day precisions in terms of percent relative standard deviation of retention times and peak areas were 0.07-3.99 for rebaudioside A, and 0.11-6.94 for stevioside. Accuracy of the purposed methods for rebaudioside A and stevioside in terms of percent recovery was in the range 92.0-101.3. The proposed method could be applied to determine rebaudioside A and stevioside in beverage and stevia samples. Keywords : rebaudioside A, stevioside, stevia, high-performance liquid chromatographyReferences
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Molina-Calle, M., Sánchez De Medina, V., Delgado De La Torre, M.P., Priego-Capote, F., and Luque De Castro, M.D. (2016). Development and application of a quantitative method based on LC-QqQ MS/MS for determination of steviol glycosides in Stevia leaves. Talanta, 154, 263–269.
Notification of the Ministry of Public Health (No. 381) (2016) Food additives. Retrieved June 30, 2018, from http://food.fda.moph.go.th/law/data/announ_ moph/P381.pdf (in Thai)
Pavlíček, V., and Tůma, P. (2017). The use of capillary electrophoresis with contactless conductivity detection for sensitive determination of stevioside and rebaudioside A in foods and beverages. Food Chemistry, 219, 193–198.
Rieck, U.W, Lankes, C., Wawrzun, A., and Wüst, M. (2010). Improved HPLC method for the evaluation of the major steviol glycosides in leaves of Stevia rebuadiana. European food Research and Technology, 231, 581-588.
Vaněk, T., Nepovím, A., and Valíček, P. (2001). Determination of stevioside in plant material and fruit teas. Journal of Food Composition and Analysis, 14, 383–388.
Bovanová, L., Brandšteterová, E., and Baxa, S. (1998). HPLC determination of stevioside in plant material and food samples. Zeitschrift Für Lebensmitteluntersuchung Und -Forschung A, 207, 352–355.
Gardana, C., Scaglianti, M., and Simonetti, P. (2010). Evaluation of steviol and its glycosides in stevia rebaudiana leaves and commercial sweetener by ultra-high-performance liquid chromatography-mass spectrometry. Journal of Chromatography A, 1217, 1463–1470.
Gasmalla, M. A. A., Yang, R. and Hua, X. (2014). Stevia rebaudiana Bertoni: An alternative sugar replacer and its application in food lndustry. Food Engineering Reviews, 6, 150–162.
Kim, N.-C. and Kinghorn, A. D. (2002). Highly sweet compounds of plant origin. Archives of Pharmacal Research, 25, 725–746.
Liu, J., and Li, S.F.Y. (1995). Separation and determination of stevia sweeteners by capillary electrophoresis and high performance liquid chromatography. Journal of liquid chromatography, 18(9), 1703-1719.
Martono, Y., Riyanto, S., Rohman, A., and Martono, S. (2016). Improvement method of fast and isocratic RP-HPLC analysis of major diterpene glycoside from Stevia rebaudiana leaves. In AIP Conference Proceedings. (pp. 1-8). American Institute of Physics: AIP Publishing.
Molina-Calle, M., Sánchez De Medina, V., Delgado De La Torre, M.P., Priego-Capote, F., and Luque De Castro, M.D. (2016). Development and application of a quantitative method based on LC-QqQ MS/MS for determination of steviol glycosides in Stevia leaves. Talanta, 154, 263–269.
Notification of the Ministry of Public Health (No. 381) (2016) Food additives. Retrieved June 30, 2018, from http://food.fda.moph.go.th/law/data/announ_ moph/P381.pdf (in Thai)
Pavlíček, V., and Tůma, P. (2017). The use of capillary electrophoresis with contactless conductivity detection for sensitive determination of stevioside and rebaudioside A in foods and beverages. Food Chemistry, 219, 193–198.
Rieck, U.W, Lankes, C., Wawrzun, A., and Wüst, M. (2010). Improved HPLC method for the evaluation of the major steviol glycosides in leaves of Stevia rebuadiana. European food Research and Technology, 231, 581-588.
Vaněk, T., Nepovím, A., and Valíček, P. (2001). Determination of stevioside in plant material and fruit teas. Journal of Food Composition and Analysis, 14, 383–388.
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2018-07-05
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