O Que Glicosdeos De Esteviol?

Los glucósidos de esteviol son compuestos químicos que se aíslan de las hojas de Stevia rebaudiana, una planta que crece en la América del Sur y ha sido utilizada por los indígenas durante cientos de años. La estructura química de los glucósidos de esteviol es estable y no sufre cambios durante el proceso de extracción y purificación – por lo que cabe suponer que los glucósidos de esteviol son un producto completamente natural. Numerosas pruebas sensoriales indican que los glucósidos de esteviol son incluso hasta 300 veces más dulces que la sacarosa convencional. Desde los años 70, se utilizan como aditivo alimentario en Japón. En los Estados Unidos, han sido marcados por la FDA con el símbolo GRAS (Generally Recognized As Safe) como completamente seguros. En 2011, la Unión Europea los aprobó como aditivo alimentario con el símboloE-960.

The residual dextrin is mainly removed from the reaction mixture by the use of an adsorption/desorption column that can remove all dextrin to a typical lower level of up to 4% by weight (Documentation provided to EFSA n.5).

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The Panel noted that, in the absence of analytical standards for all the different glycosylated steviol glycosides (including all of the possible regioisomers), this method can only provide semiquantitative results.

The safety of steviol glycosides as a food additive was evaluated by EFSA in 2010 and an acceptable daily intake (ADI) of 4 mg/kg body weight (bw) (expressed as steviol equivalents) per day was established.

Appendix A – Typical components of glucosylated steviol glycoside preparations (Documentation provided to EFSA n. 1)

Although not conclusive, the Panel found this explanation to be reasonable. The Panel noted, however, that the relevance of this last study to the stability of glucosylated steviol glycosides was limited due to the short heating period and temperature range employed (e.g. the latter not being representative of baking processes).

Morphology of Stevia rebaudiana plants at full vegetative growth (a) and at the beginning of flowering (b) cultivated in the open field under temperate climate conditions of Southern Poland or cultivated in the growth chamber under influence of different stress inducing factors (c) (source: unpublished photos by Marta Libik-Konieczny)

Stevia rebaudiana (Bertoni) biology and agronomy

Existen diferentes tipos de glicósidos de esteviol, también llamados esteviósidos y no son un aditivo alimentario nuevo, pues ya han sido aprobados en diferentes países y continentes y se estima que más de 150 millones de personas de diferentes partes del mundo consumen a diario estos edulcorantes como componentes adicionados a diversos alimentos industriales.

Glucosylated steviol glycosides’ preparations were described by the applicant as a mixture of glucosylated steviol glycosides, containing 1–20 additional glucose units bound to the parent steviol glycoside via α‐(1–4) linkages. The Panel noted that the definition of the food additive in the proposed specifications does not include any value for the additional glucose units and that the mixtures on which data have been provided contain, on average, only three or four additional glucose units. The Panel made a conservative assumption that the GSG may contain up to 10 additional glucose units on average in its evaluation and considered that information on the number of additional glucose units (1–20) should be added to the proposed definition of the food additive in the specifications.

Effect of SGs on low-density lipoprotein cholesterol (LDL-C, mg/dL) CI: confidence interval; MD: mean difference; RCTs: randomized controlled trials; SD: standard deviation.

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1. Introduction

Since the glucosylated steviol glycosides are expected to readily undergo stepwise hydrolysis in the GI tract forming steviol, the release of glucose can be estimated as follows.

For the GSG‐RA mixture, by considering that it is composed only of Rebaudioside A (four glucose units/molecule) and that enzymatic glucosylation resulted in the addition of 2.95 glucose units/molecule, a maximum exposure of 4.3 mg/kg bw per day (expressed as steviol equivalents) corresponds to a maximum exposure to steviol (MW 318) of 13.52 µmoles/kg bw per day. Since each mole of steviol equivalents yields 6.95 moles of glucose (MW 180), the maximum exposure to glucosylated steviol glycoside‐derived glucose would be 16.9 mg/kg bw per day.

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The purified steviol glycosides (≥ 95%), extracted from the leaves of the Stevia rebaudiana Bertoni plant, are enzymatically modified such that additional glucose moieties are conjugated to the parent steviol glycosides structure via α‐(1–4) linkages. Glucosylated steviol glycoside preparations consist of not less than 95% total steviol glycosides, made up of any combination of glucosylated steviol glycosides of different molecular weights as well as any remaining unreacted steviol glycosides (parent steviol glycosides). According to the applicant, the percentage of steviol glycosides containing additional molecules of glucose is between 80% and 92% w/w and the parent glycosides between 5% and 15% w/w (Documentation provided to EFSA n. 1, 4).

The Panel considered that rather than having a non‐definite list of components, as proposed by the applicant, the assay value should cover all steviol glycosides falling under the definition of E 960a along with their glucosylated derivatives.

The purified steviol glucosides’ extract is mixed with the liquified tapioca starch along with the CGTase. The reaction mixture is incubated at 60°C for 48 h to generate glucosylated steviol glycosides and dextrin (residual hydrolysed starch). The enzyme is then inactivated by heating at 100°C for 15 min and removed in the subsequent purification steps.

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The present evaluation is based on the data on steviol glycosides in a newly submitted dossier by the applicant and additional information submitted by the applicant during the assessment process either spontaneously or following request from EFSA.

European Food Safety Authority, Panel on Food Additives and Nutrient Sources added to Food. Scientific opinion on the safety of steviol glycosides for the proposed uses as a food additive. EFSA Journal, 8(4):1537. 2010. [3. Biological and toxicological data (pg 20); 3.1. Absorption, distribution, metabolism and excretion (pg 20); 3.1.1. In vitro studies (pg 20);3.1.2. In vivo studies (pg 20)]. www.efsa.europa.eu/en/efsajournal/pub/1537.htm

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According to the applicant, the approach including adsorption chromatography is preferred because it estimates the total steviol equivalency of a glucosylated steviol glycoside preparation which is needed to estimate the use of glucosylated steviol glycosides in different food and beverage applications, where the additive level is defined/expressed as steviol equivalents.

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Glucosylated steviol glycosides are prepared in two production stages. Steviol glycosides mixtures – enriched individual steviol glycosides (e.g. high Rebaudioside A or Rebaudioside D content (> 95%)) or mixtures enriched in two or more individual steviol glycosides – are isolated from dried leaves of S. rebaudiana by hot water extraction and different purifications steps. The purified steviol glycosides extract undergoes enzymatic treatment to achieve glucosylation via α‐(1‐4) linkages of the steviol glycosides. A glucose donor, such as tapioca starch (extracted from Cassava roots), is treated with the enzyme CGTase (EC 2.4.1.19), derived from a non‐genetically modified strain of Anoxybacillus caldiproteolyticus. CGTase (EC 2.4.1.19) is also used to catalyse the intermolecular glucosylation whereby the transfer occurs of α‐glucosyl units from the starch onto the 4‐hydroxy position of a glucosyl moiety on the steviol glycoside (trans‐α‐1,4‐glucosylation). The enzyme is inactivated by heating and removed by treatment with activated carbon. The dextrin by‐product can be removed from the mixture.

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European Commission Regulation (EU) No 1131/2011 of 11 November 2011 amending Annex II to Regulation (EC) No 1333/2008 of the European Parliament and of the Council with regard to steviol glycosides. Official Journal of the European Union. December 11, 2011. Retrieved June 13, 2013: http://eurlex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2011:295:0205:0211:EN:PDF

Glucosylated steviol glycosides’ preparations were described by the applicant as a mixture of glucosylated steviol glycosides, containing 1–20 additional glucose units bound to the parent steviol glycoside via α‐(1–4) linkages. The Panel noted that the definition of the food additive in the proposed specifications does not include any value for the additional glucose units and that the mixtures on which data have been provided contain, on average, only three or four additional glucose units. The Panel made a conservative assumption that the GSG may contain up to 10 additional glucose units on average in its evaluation and considered that information on the number of additional glucose units (1–20) should be added to the proposed definition of the food additive in the specifications.