Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method

Joannes Martinus Cornelius Geuns

doi:doi:10.11648/j.ijnfs.20180705.13

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Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method

Joannes Martinus Cornelius Geuns

Published in International Journal of Nutrition and Food Sciences (Volume 7, Issue 5)

Received: 29 August 2018 Accepted: 10 September 2018 Published: 11 October 2018

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Abstract

To optimise steviol glycoside analysis, several round-robin tests were organised by the European Stevia Association (EUSTAS). Seventeen laboratories participated in the testing. Only 8 laboratories have sent their results. In the first round-robin testing, 2 samples were analysed. The first sample had a purity of 96.2%. The second sample was a 4/5 dilution with NaHCO₃ of sample 1. This way, the drying process itself could be checked. The purity of sample 2 was 82.35%. The reported purities of sample 1 varied between 79.8 and 96.2%, those of sample 2 varied between 58.1 and 81.8%. The drying of sample 2 showed that weight loss was between 4.9 and 12.7%, demonstrating that not all laboratories dried the sample to a constant weight. In a second round-robin testing, the purity of the sample was 91.1%. The reported purities of the sample varied between 82.74 and 95.86%. About 3% purified RebB was added to the sample to check the quality of the analysis of this compound possessing a carboxylic group. The samples contained the following steviol glycosides: Reb D, Reb E, Reb A, ST, Reb F, Reb C, Dul A, Reb G, Rub, Reb B, SB and SM (1 lab). No SV was detected. The number of SVgly analysed in the different laboratories varied between 4 and 11. One lab only analysed ST and Reb A and gave a percentage composition of these compounds. To improve the accuracy of analysis, different suggestions are made, such as controlling the drying process of samples and standards, purity of standards, injection of sufficient material and use of solvent gradients to shorten the run time and to reduce integration errors. The results of this second round-robin tests are better than those of the first one.

Published in	International Journal of Nutrition and Food Sciences (Volume 7, Issue 5)
DOI	10.11648/j.ijnfs.20180705.13
Page(s)	173-179
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2018. Published by Science Publishing Group

Keywords

Steviol Glycosides, Round-robin Testing, Validation of Analytical Methods, HPLC

References

[1]	Geuns J.M.C. and Struyf T.: EUSTAS Round-Robin Testing of Steviol Glycosides pp. 35-48 in Proceedings of the 3^rd EUSTAS Stevia Symposium “Stevia in Europe” July 1^st – 2^nd 2009, KULeuven, Belgium Ed.: Jan M.C. Geuns, ISBN: 978-90-742-53079.
[2]	Geuns J.M.C.: Stevia and steviol glycosides. (2010) Euprint Ed., Parkbosstraat 3, 3001 Heverlee, Belgium, ISBN: 9789074253116, pp. 307.
[3]	JECFA (2010) Steviol glycosides. FAO JECFA Monograph 10.
[4]	Scaglianti M., C. Gardana, P.G. Pietta and G.M. Ricchiuto (2008) Analysis of the main Stevia rebaudiana sweeteners and their aglycone Steviol by a validated LC-DAD-ESI-MS method. pp. 45-58 in Proceedings of the EUSTAS Stevia Symposium, June 27^th 2008, KULeuven, Belgium Ed.: Jan M.C. Geuns, ISBN: D/2008/6045/50.
[5]	Zimmermann B.F. and U. Wölwer-Rieck (2011) HILIC columns for the analysis of steviol glycosides. pp. 147-152 in Proceedings of the 5^th EUSTAS Stevia Symposium Stevia, Breakthrough in Europe, June 28-29, 2011, KULeuven, Belgium Ed.: Jan M.C. Geuns, ISBN: 978-90-742-53192; D/2011/6045/038.
[6]	Zimmermann B.F. (2018) Beaming steviol glycoside analysis into the next dimension. Food Chemistry, 241, 150-153.
[7]	Chatervedula, V. and Zamora J. (2014) Reversed-phase HPLC analysis of steviol glycosides isolated from Stevia rebaudiana Bertoni. Food and Nutrition Sciences 5, 1711-1716.
[8]	Tada A., Ishizuki K., Mikami H., Hirao Y., Fujita L. et al. (2013) Improvement of the assay methodo for steviol glycosides in the JECFA specifications. American Journal of Analytical Chemistry 4, 190-196.
[9]	Soufi S., D’Urso G., Pizza C., Rezgui S, Bettaieb T and Montoro P (2016) Steviol glycosides targeted analysis in leaves of Stevia rebaudiana (Bertoni) from plants cultivated under chilling stress conditions. Food Chemistry 190(C), 572-580.
[10]	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(C), 263-269.
[11]	Geuns J.M.C. (2008) Analysis of Steviol glycosides: validation of the methods. pp. 59-78 in Proceedings of the EUSTAS Stevia Symposium, June 27^th 2008, KULeuven, Belgium Ed.: Jan M.C. Geuns, ISBN: D/2008/6045/50.
[12]	Struyf T., Chandia N.P., De Borggraeve W., Dehaen W. and Geuns.J.M.C.: Preparation of pure standards of steviol glycosides. Identification of steviol glycosides by LC-MS and NMR. pp. 29-44 in Proceedings of the EUSTAS Stevia Symposium, June 27^th 2008, KULeuven, Belgium Ed.: Jan M.C. Geuns, ISBN: 9789074253-031; D/2008/6045/50.
[13]	Kumari N. and Kumar S. (2017) Chemistry and analytical techniques for ent-kaurene-glycosides of Stevia rebaudiana Bertoni – A review. Journal of Applied and Natural Sciences 9 (4), 2114-2126.
[14]	Ceunen S. and Geuns J.M.C. (2013) Influence of photoperiodism on the spatio-temporal accumulation of steviol glycosides in Stevia rebaudiana (Bertoni). Plant Science 198, 72-82.
[15]	EFSA (2011) EU Commission. Commission Regulation (EU) No. 1131/2011. Official J. Eur. Union 2011, L295, 205−211.

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Joannes Martinus Cornelius Geuns. (2018). Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method. International Journal of Nutrition and Food Sciences, 7(5), 173-179. https://doi.org/10.11648/j.ijnfs.20180705.13

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ACS Style

Joannes Martinus Cornelius Geuns. Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method. Int. J. Nutr. Food Sci. 2018, 7(5), 173-179. doi: 10.11648/j.ijnfs.20180705.13

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AMA Style

Joannes Martinus Cornelius Geuns. Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method. Int J Nutr Food Sci. 2018;7(5):173-179. doi: 10.11648/j.ijnfs.20180705.13

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@article{10.11648/j.ijnfs.20180705.13,
  author = {Joannes Martinus Cornelius Geuns},
  title = {Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method},
  journal = {International Journal of Nutrition and Food Sciences},
  volume = {7},
  number = {5},
  pages = {173-179},
  doi = {10.11648/j.ijnfs.20180705.13},
  url = {https://doi.org/10.11648/j.ijnfs.20180705.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijnfs.20180705.13},
  abstract = {To optimise steviol glycoside analysis, several round-robin tests were organised by the European Stevia Association (EUSTAS). Seventeen laboratories participated in the testing. Only 8 laboratories have sent their results. In the first round-robin testing, 2 samples were analysed. The first sample had a purity of 96.2%. The second sample was a 4/5 dilution with NaHCO3 of sample 1. This way, the drying process itself could be checked. The purity of sample 2 was 82.35%. The reported purities of sample 1 varied between 79.8 and 96.2%, those of sample 2 varied between 58.1 and 81.8%. The drying of sample 2 showed that weight loss was between 4.9 and 12.7%, demonstrating that not all laboratories dried the sample to a constant weight. In a second round-robin testing, the purity of the sample was 91.1%. The reported purities of the sample varied between 82.74 and 95.86%. About 3% purified RebB was added to the sample to check the quality of the analysis of this compound possessing a carboxylic group. The samples contained the following steviol glycosides: Reb D, Reb E, Reb A, ST, Reb F, Reb C, Dul A, Reb G, Rub, Reb B, SB and SM (1 lab). No SV was detected. The number of SVgly analysed in the different laboratories varied between 4 and 11. One lab only analysed ST and Reb A and gave a percentage composition of these compounds. To improve the accuracy of analysis, different suggestions are made, such as controlling the drying process of samples and standards, purity of standards, injection of sufficient material and use of solvent gradients to shorten the run time and to reduce integration errors. The results of this second round-robin tests are better than those of the first one.},
 year = {2018}
}

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TY - JOUR
T1 - Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method
AU - Joannes Martinus Cornelius Geuns
Y1 - 2018/10/11
PY - 2018
N1 - https://doi.org/10.11648/j.ijnfs.20180705.13
DO - 10.11648/j.ijnfs.20180705.13
T2 - International Journal of Nutrition and Food Sciences
JF - International Journal of Nutrition and Food Sciences
JO - International Journal of Nutrition and Food Sciences
SP - 173
EP - 179
PB - Science Publishing Group
SN - 2327-2716
UR - https://doi.org/10.11648/j.ijnfs.20180705.13
AB - To optimise steviol glycoside analysis, several round-robin tests were organised by the European Stevia Association (EUSTAS). Seventeen laboratories participated in the testing. Only 8 laboratories have sent their results. In the first round-robin testing, 2 samples were analysed. The first sample had a purity of 96.2%. The second sample was a 4/5 dilution with NaHCO3 of sample 1. This way, the drying process itself could be checked. The purity of sample 2 was 82.35%. The reported purities of sample 1 varied between 79.8 and 96.2%, those of sample 2 varied between 58.1 and 81.8%. The drying of sample 2 showed that weight loss was between 4.9 and 12.7%, demonstrating that not all laboratories dried the sample to a constant weight. In a second round-robin testing, the purity of the sample was 91.1%. The reported purities of the sample varied between 82.74 and 95.86%. About 3% purified RebB was added to the sample to check the quality of the analysis of this compound possessing a carboxylic group. The samples contained the following steviol glycosides: Reb D, Reb E, Reb A, ST, Reb F, Reb C, Dul A, Reb G, Rub, Reb B, SB and SM (1 lab). No SV was detected. The number of SVgly analysed in the different laboratories varied between 4 and 11. One lab only analysed ST and Reb A and gave a percentage composition of these compounds. To improve the accuracy of analysis, different suggestions are made, such as controlling the drying process of samples and standards, purity of standards, injection of sufficient material and use of solvent gradients to shorten the run time and to reduce integration errors. The results of this second round-robin tests are better than those of the first one.
VL - 7
IS - 5
ER -

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Joannes Martinus Cornelius Geuns

Laboratory of Functional Biology, KU Leuven, Leuven, Belgium

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APA Style

Joannes Martinus Cornelius Geuns. (2018). Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method. International Journal of Nutrition and Food Sciences, 7(5), 173-179. https://doi.org/10.11648/j.ijnfs.20180705.13

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ACS Style

Joannes Martinus Cornelius Geuns. Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method. Int. J. Nutr. Food Sci. 2018, 7(5), 173-179. doi: 10.11648/j.ijnfs.20180705.13

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AMA Style

Joannes Martinus Cornelius Geuns. Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method. Int J Nutr Food Sci. 2018;7(5):173-179. doi: 10.11648/j.ijnfs.20180705.13

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@article{10.11648/j.ijnfs.20180705.13,
  author = {Joannes Martinus Cornelius Geuns},
  title = {Inter-Laboratory Analysis of Steviol Glycosides by an External Standard Method},
  journal = {International Journal of Nutrition and Food Sciences},
  volume = {7},
  number = {5},
  pages = {173-179},
  doi = {10.11648/j.ijnfs.20180705.13},
  url = {https://doi.org/10.11648/j.ijnfs.20180705.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijnfs.20180705.13},
  abstract = {To optimise steviol glycoside analysis, several round-robin tests were organised by the European Stevia Association (EUSTAS). Seventeen laboratories participated in the testing. Only 8 laboratories have sent their results. In the first round-robin testing, 2 samples were analysed. The first sample had a purity of 96.2%. The second sample was a 4/5 dilution with NaHCO3 of sample 1. This way, the drying process itself could be checked. The purity of sample 2 was 82.35%. The reported purities of sample 1 varied between 79.8 and 96.2%, those of sample 2 varied between 58.1 and 81.8%. The drying of sample 2 showed that weight loss was between 4.9 and 12.7%, demonstrating that not all laboratories dried the sample to a constant weight. In a second round-robin testing, the purity of the sample was 91.1%. The reported purities of the sample varied between 82.74 and 95.86%. About 3% purified RebB was added to the sample to check the quality of the analysis of this compound possessing a carboxylic group. The samples contained the following steviol glycosides: Reb D, Reb E, Reb A, ST, Reb F, Reb C, Dul A, Reb G, Rub, Reb B, SB and SM (1 lab). No SV was detected. The number of SVgly analysed in the different laboratories varied between 4 and 11. One lab only analysed ST and Reb A and gave a percentage composition of these compounds. To improve the accuracy of analysis, different suggestions are made, such as controlling the drying process of samples and standards, purity of standards, injection of sufficient material and use of solvent gradients to shorten the run time and to reduce integration errors. The results of this second round-robin tests are better than those of the first one.},
 year = {2018}
}

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