Identification and Analysis of Steviol Glycosides in Stevia Rebaudiana Leaves by HPLC-MS

doi:10.11924/j.issn.1000-6850.casb16090028

Chin Agric Sci Bull ›› 2017, Vol. 33 ›› Issue (30) : 43-50. DOI: 10.11924/j.issn.1000-6850.casb16090028

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Abstract

To improve the measurement method of steviol glycosides (SGs) in Stevia leaves, several SGs standards and leaf extracts of‘Xinfeng 3’were analyzed and determined by using high performance liquid chromatography-mass spectrometer (HPLC-MS). And the retention time and order of 12 different SGs were identified, among which three (rubusoside, steviolbioside and rebaudioside E) were first reported. Based on the rebaudiodside A standard curve and HPLC fingerprints map of SGs, the composition of SGs in‘Xinfeng 3’ leaves was determined. Totally nine kinds of SGs including stevioside, rebaudioside A, B, C, D, E and F, rubusoside and steviolbioside were identified. The rebaudioside D and E were presented in trace amount. This study provides not only detailed ultraviolet-map for further detection of SGs, but also a reference method for finding and identifying new SGs.

Key words

Stevia rebaudiana ; steviol glycosides; HPLC-MS; amio chromatographic column; rebaudioside A

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Identification and Analysis of Steviol Glycosides in Stevia Rebaudiana Leaves by HPLC-MS[J]. Chinese Agricultural Science Bulletin. 2017, 33(30): 43-50 https://doi.org/10.11924/j.issn.1000-6850.casb16090028

References

[1] Yadav A K, Singh S, Dhyani D, et al. A review on the improvement of stevia [Stevia rebaudiana (Bertoni)] [J]. Canadian Journal of Plant Science, 2011, 91(1): 1-27.
[2] 王飞生, 叶荣飞, 闵建. 甜菊糖苷的特性及应用 [J]. 中国调味品, 2009, 10 (34): 91-95.
[3] Sakamoto I, Kohda H, Murakami K, et al. Quantitative analysis of stevioside (author's transl) [J]. Yakugaku zasshi: Journal of the Pharmaceutical Society of Japan, 1975, 95(12): 1507.
[4] Kohda H, Kasai R, Yamasaki K, et al. New sweet diterpene glucosides from Stevia rebaudiana [J]. Phytochemistry, 1976, 15(6): 981-983.
[5] Chaturvedula V S P, Prakash I. Diterpene glycosides from Stevia rebaudiana [J]. Journal of Medicinal Plants Research, 2011, 5(19): 4838-4842.
[6] Mantovaneli I C C, Ferretti E C, Sim?es M R, et al. The effect of temperature and flow rate on the clarification of the aqueous stevia-extract in a fixed-bed column with zeolites [J]. Brazilian Journal of Chemical Engineering, 2004, 21(3): 449-458.
[7] de Oliveira B H, Packer J F, Chimelli M, et al. Enzymatic modification of stevioside by cell-free extract of Gibberella fujikuroi [J]. Journal of biotechnology, 2007, 131(1): 92-96.
[8] Morlock G E, Meyer S, Zimmermann B F, et al. High-performance thin-layer chromatography analysis of steviol glycosides in Stevia formulations and sugar-free food products, and benchmarking with (ultra) high-performance liquid chromatography [J]. Journal of Chromatography A, 2014, 1350: 102-111.
[9] Shah R, De Jager L S, Begley T H. Simultaneous determination of steviol and steviol glycosides by liquid chromatography-mass spectrometry [J]. Food Additives & Contaminants: Part A, 2012, 29(12): 1861-1871.
[10] Shafii B, Vismeh R, Beaudry R, et al. Large-scale profiling of diterpenoid glycosides from Stevia rebaudiana using ultrahigh performance liquid chromatography/tandem mass spectrometry [J]. Analytical and bioanalytical chemistry, 2012, 403(9): 2683-2690.
[11] Jamaluddin Ahmed M, Smith R M. Determination of stevioside by high‐performance liquid chromatography with pulsed amperometric detection [J]. Journal of separation science, 2002, 25(3): 170-172.
[12] Bililign T, Moore J C, Tan S, et al. Development and validation of a reversed-phase high-performance liquid chromatography method for routine identification and purity assessment of high-purity steviol glycoside sweeteners [J]. Journal of agricultural and food chemistry, 2014, 62(6): 1384-1389.
[13] Montoro P, Molfetta I, Maldini M, et al. Determination of six steviol glycosides of Stevia rebaudiana (Bertoni) from different geographical origin by LC–ESI–MS/MS [J]. Food chemistry, 2013, 141(2): 745-753.
[14] Espinoza M I, Vincken J P, Sanders M, et al. Identification, quantification, and sensory characterization of steviol glycosides from differently processed Stevia rebaudiana commercial extracts [J]. Journal of agricultural and food chemistry, 2014, 62(49): 11797-11804.
[15] Molina-Calle M, de Medina V S, de la Torre M P D, et al. Development and application of a quantitative method based on LC–QqQ MS/MS for determination of steviol glycosides in Stevia leaves [J]. Talanta, 2016, 154: 263-269.
[16] Jamaluddin Ahmed M, Smith R M. Determination of stevioside by high‐performance liquid chromatography with pulsed amperometric detection [J]. Journal of separation science, 2002, 25(3): 170-172.
[17] Jan M.C. Genus Stevia and steviol glycosides [M]. 北京：中国轻工业出版社，2013: 41.
[18] Montoro P, Molfetta I, Maldini M, et al. Determination of six steviol glycosides of Stevia rebaudiana (Bertoni) from different geographical origin by LC–ESI–MS/MS[J]. Food chemistry, 2013, 141(2): 745-753.
[19] 谢莹，陈全斌，罗达伟, 等. 高纯度甜茶苷抗疲劳及免疫调节功能研究 [J].时珍国医国药, 2010, 21(6):1421-1422.
[20] 孙传范，李进伟. 甜菊糖苷研究进展[J]. 食品科学，2010, 1(31)：338-340.
[21] 王德骥. 关于甜菊糖苷的甜度、甜味和苦涩后味的成因机理 [J]. 中国食品添加剂, 2007：46-53.
[22] Kwang-He K. Physio-ecological studies on Stevia (Stevia rebaudiana Bertoni)[J]. Korean Journal of Crop Science, 1981, 26(1): 69-89.
[23] Bondarev N I, Sukhanova M A, Reshetnyak O V, et al. Steviol glycoside content in different organs of Stevia rebaudiana and its dynamics during ontogeny[J]. Biologia Plantarum, 2003, 47(2): 261-264.
[24] Bondarev N I, Reshetnyak O V, Bondareva T A, et al. Distribution and dynamics of diterpene glycosides contents in stevia rebaudiana bertoni intact plants[J]. Biotechnology, 2012 (5): 50-54.
[25] Ceunen S, Geuns J M C. Influence of photoperiodism on the spatio-temporal accumulation of steviol glycosides in Stevia rebaudiana (Bertoni)[J]. Plant science, 2013, 198: 72-82.
[26] Modi A R, Raj S, Kanani P, et al. Analysis of differentially expressed genes involved in stevioside biosynthesis in cultures of Stevia rebaudiana Bertoni treated with steviol as an immediate precursor[J]. Journal of plant growth regulation, 2014, 33(3): 481-488.
[27] Ceunen S, Geuns J M C. Steviol glycosides: chemical diversity, metabolism, and function[J]. Journal of natural products, 2013, 76(6): 1201-1228.