Stevia - a non-caloric sweetener of natural origin

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September 2015

Steviol glycosides are a group of high-potency, zero-calorie sweeteners isolated from leaves of Stevia rebaudiana. Permitted as sweeteners in the EU since December 2011, steviol glycosides offer an opportunity to reduce the added sugar in a range of foods and beverages without loss of sweetness or reliance on synthetic sweeteners. The origin of the glycosides is described, together with key aspects of their sweetness intensity and quality. EU regulations governing the use of these sweeteners are outlined and formulation advice and guidelines on stability of the glycosides  provided.


ADI              Acceptable Daily Intake
EFSA           European Food Safety Authority
EU               European Union
EVIRA          Elintarviketurvallisuusvirasto Finnish Food Safety Authority
FDA             Food and Drugs Administration (of the United States)
FSA             Food Standards Agency (UK)
GRAS          Generally Recognized As Safe
HPS             High Potency Sweetener
JECFA         Joint Expert Committee (of the World Health Organization and the Food and Agriculture Organization of the United Nations) on Food Additives
SE               Sucrose Equivalent
QS               Quantum Satis


Until recently, synthetic high-potency sweeteners (HPS) were the only practical solution to the problem of removing sugars from food and drinks without loss of sweetness.

Steviol glycosides (E960) were added to the European Union (EU) list of permitted sweeteners in 2011. This meant that, for the first time, there was available a non-caloric HPS, derived from nature, that had a taste quality good enough for it to be a significant source of sweetness in soft drinks and other products. This statement considers the origin, taste quality, regulation and practical application of steviol glycosides.

Botanical Source

The shrub Stevia rebaudiana is a member of the Asteraceae and native to Paraguay, where the leaves have a history of use as a sweetener for over a century (Kinghorn 2002). Uniquely, the plant secretes in its leaves a group of structurally-related high-potency sweeteners, the steviol glycosides. All are ent-kaurene diterpenoid glycosides with a common core of steviol (ent-13-hydroxykaur-16-en-19-oic acid - see Figure 1). The most abundant of these molecules in the wild-type plant is stevioside, but the less-plentiful rebaudioside A (reb A) is widely held to be the better tasting of the two (Fry 2012; Phillips 1987). There are many other glycosides in the leaf, but generally at minor to trace concentrations. Because of the taste preference for reb A, conventional plant breeding programmes have been directed over many years to increasing the yield of this compound. Dried leaves of Stevia rebaudiana typically contain about 7-15% by weight steviol glycosides (Carakostas et al. 2011). Of this, 40% might be reb A in a modern cultivar (Morita and Bu 2000a, 2000b; Morita et al. 2009).

Stevia is widely cultivated in South America, but much of the world's commercial supply currently originates in Asia. There has also been interest in growing the crop in numerous other locations, including Canada, Australia, the USA, Southern Mediterranean countries and Africa.

The glycosides are extracted by steeping dry leaves in warm water, filtering the extract free of solid material, and purifying the resultant solution by a combination of chromatography and crystallisation from water-alcohol solutions. This results in a crude extract comprising a mixture of glycosides in variable proportions, generally containing predominantly stevioside and rebaudioside A. Such extracts may be used directly, but are normally further refined by additional recrystallisation to create ingredient-grade HPS that meet the purity requirements of Western markets.

Commercial glycoside products

The first ingredient-grade product launched in Western markets was essentially pure reb A. Lower-cost extracts containing less reb A quickly followed. These are often described by their reb A content such as “RA80” or “RA50”, meaning a minimum of 80 or 50% reb A respectively. Contrary to expectations, extracts that were not pure reb A but which had high reb A content (say around RA70 and above) were found to be of similar sweetness potency to pure reb A but with a superior taste quality. This observation inspired far-reaching investigations of the taste qualities of individual glycosides and their interactions when blended. The insights thus afforded are the basis of a new generation of stevia ingredients where leaf extracts are carefully blended to maximise potency while reducing unwanted side-tastes such as bitterness or licorice.

Alongside these developments, suppliers have looked to earlier Japanese studies involving enzymic modification of steviol glycosides to improve taste quality. It is a general rule that the greater the number of sugar groups attached to the steviol nucleus, the better the sweetness quality. Enzyme modification increases the number of substituent sugar groups by incubating a low-cost extract (usually high in stevioside) with a glucosyl transferase enzymes and a source of glucose. The original plant glycosides emerge modified by the addition of one to three (or more) extra glucose residues. The exact glycoside composition that results is often indeterminate. The material usually also contains some unreacted glucose and a small quantity of unreacted feedstock glycosides. Enzyme modified stevia does not meet EU regulatory requirements for sweetener-grade steviol glycosides and is primarily sold to flavour houses as a flavouring and sweetness enhancer.

Further study of minor glycosides from the leaf has focused attention on compounds such as reb D and reb M. These have a much higher quality of sweetness than reb A, or even controlled blends of leaf extracts, and have excited much interest. The drawback of such minor glycosides is that they exist in the leaf at such low concentrations that their isolation from this source is prohibitively costly. Work is afoot to make such glycosides commercially available by using a fermentation process based on genetically modified yeast. The glycosides produced are identical to those in the stevia leaf and would be regarded as natural for regulatory purposess in the USA - not the case in the EU. Accordingly, while such products may appear commercially in the USA in 2016, their use in the EU is likely to be more delayed.

Safety and regulation

Modern investigations, including human studies on safety, metabolism and intake, fully support the safety of stevia sweeteners.

Stevia has been the subject of biological and toxicological investigations for more than 50 years. The earliest of these studies were conducted with crude or poorly defined test materials. These historical results were both unacceptable to modern toxicologists, and the source of some confusion about actual safety. Both matters were eventually resolved by work conducted with fully characterised, high-purity reb A extracts (Brusik 2008). This led to the USA FDA issuing a letter of “no objection” to the designation of reb A as “Generally Recognized As Safe” (GRAS) in December 2008. The GRAS designation was subsequently accepted for extracts containing other steviol glycosides (Carakostas et al. 2008).

Elsewhere, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) thoroughly reviewed the data on steviol glycosides and concluded that they are safe for use in food and beverages for people of all ages and populations. JECFA first set tentative purity specifications in 2004 and published a final version in 2010 (JECFA 2010). An Acceptable Daily Intake (ADI) of 4 mg/kg body weight/day (expressed as steviol) was established in 2008 and remains current. This compares with 40 mg/kg for aspartame, 5 mg/kg for saccharin and 15 mg/kg sucralose. To put these figures in context, 4 mg/kg as steviol equates to 12 mg/kg of rebaudioside A.

In 2010, the European Food Safety Authority (EFSA) concluded that steviol glycosides are safe and established the same ADI as JECFA. This resulted in a European regulation permitting steviol glycosides in a range of food and drink from December 2011 (European Commission, 2011).

Today, there are a large number of studies on stevia in the scientific literature that corroborate the safety conclusions of regulatory agencies around the globe. Table 1 includes a list of safety reviews and regulatory approvals that have enabled growth of the stevia sweetener market.


Table 1 Global safety reviews for steviol glycosides






Joint FAO/WHO Expert

Committee on Food Additives



Comprehensively examined the safety data and set a permanent ADI

Steviol glycosides added to the Codex Global Standard for Food Additives in 2011.




Issued official agency response letters to 11 companies who have submitted safety dossiers affirming steviol glycosides as GRAS


Food Standards Australia

New Zealand


2008, 2010

Safety review completed 2008, revised in 2010 to allow increased use levels for two food categories

(non-alcoholic beverages, ice cream)


French Food

Safety Agency



Assessed safety of rebaudioside A (a steviol glycoside)

Temporary legal approval of  97% rebaudioside A for up to a maximum of 2 years, for use in certain

food and beverages in France, published on 6 September 2009

First market in EU to approve




Scientific opinion affirming safety of steviol glycosides established an ADI

Steviol glycosides are approved for use in many other countries, including Canada, Korea, Mexico, Taiwan, China, Russia, Australia, Argentina, New Zealand, Colombia, Peru, Uruguay, Brazil, Japan and Malaysia.

EU regulatory position - composition

The EU recognises 10 steviol glycosides (European Commission, 2011) and currently requires that any commercial ingredient be a minimum 75% of rebaudioside A and/or stevioside (dry weight basis). In addition, up to 5% non-steviol glycoside material is permitted. In practice, this fraction usually comprises non-EU-recognised steviol glycosides. The remainder may be any of the recognised glycosides in any proportion.

The breadth of this specification means there is a wide range of ingredient compositions available on the market, extending from pure rebaudioside A to virtually pure stevioside, and including extracts that contain not-inconsiderable quantities of other EU-recognised steviol glycosides.

The obligatory 75% rebaudioside A/stevioside provision is proving a stumbling block in that best-tasting combinations of glycosides fall outside this limit. These blends, therefore, may not be used in the EU, although they are available in the USA. Formal requests from industry to amend the compositional requirement are likely.

EU regulatory position - use levels

The permission to use this sweetener across the EU is defined by category and addition rates limited by steviol equivalence, all outlined in Commission Regulation No. 1131/2011 (Table 2). Other categories may be added in future.


Table 2 Maximum level of steviol equivalents in specific categoriesMaximum level



Maximum steviol equivalents

(mg/l or mg/kg)


Flavoured fermented milk products


Only energy reduced products or with no added sugar

Edible Ices


Only energy reduced products or with no added sugar

Fruit and vegetables in vinegar, oil or brine


Only sweet-sour preserves of fruits and vegetables

Fruit and vegetable preparations excluding compotes


Only energy reduced

Extra jam and extra jelly as defined by Directive 2001/113/EC


Only energy reduced jams, jellies and marmalades

Jam, jellies and marmalades and sweetened chestnut puree as defined by Directive 2001/113/EC


Only energy reduced jams, jellies and marmalades

Other similar fruit or vegetable spreads


Only dried-fruit based sandwich spreads, energy-reduced or with no added sugar

Cocoa and chocolate products covered by Directive 2000/36/EC


Only energy reduced or with no added sugars

Other confectionery including breath refreshing micro sweets











Only cocoa or dried fruit based, energy reduced or with no added sugar

Only cocoa, milk, dried fruit or fat based sandwich spreads, energy reduced or with no added sugar

Only confectionery with no added sugar

Only breath-freshening micro-sweets with no added sugar

Only strongly flavoured throat pastilles with no added sugar

Chewing gum


Only with no added sugar

Decorations, coatings and fillings, except fruit based fillings



Only confectionery with no added sugar

Only cocoa or dried fruit based, energy reduced or with no added sugar

Breakfast cereals


Only cereals with a fibre content of >15%, and containing at least 20% bran, energy reduced or with no added sugar

Fine bakery wares


Only essoblaten – wafer paper

Processed fish and fishery products including molluscs and crustaceans


Only sweet-sour preserves and semi preserves of fish and marinades of fish, crustaceans and molluscs

Table Top Sweeteners



Soups and broths


Only energy reduced




Except soy-bean sauce

Only soy-bean sauce

Dietary foods for special medical purposes defined in Directive 1999/21/EC



Dietary foods for weight control; diets intended to replace total daily food intake or an individual meal



Fruit nectars as defined by Directive 2001/112/EC and veg nectars


Only energy reduced products or with no added sugar

Flavoured drinks


Only energy reduced products or with no added sugar

Beer and malt beverages


Only alcohol-free beer or with an alcohol content not exceeding 1.2% vol

Other alcoholic drinks including spirits with less than 15% alcohol and mixtures of alcoholic drinks with non-alcoholic drinks



Potato, cereal, flour or starch -based snacks



Processed nuts






Only energy reduced products or with no added sugar

Food supplements sold in solid form



Food supplements sold in liquid form



Food supplements sold in syrup type or chewable form




EU regulatory position - steviol equivalents

The reason for expressing use limits as steviol equivalent is that the steviol nucleus is the only part of the sweeteners that is of toxicological interest; the attached sugar residues are toxicologically irrelevant. Unfortunately, because the glycosides differ in structure and molecular weight, the legally-recognised glycosides each contribute different amounts of steviol (see Figure 1). As a result, manufacturers need to know the steviol equivalent of the particular mixture of glycosides they are using. From the manufacturers’ point of view it is highly desirable that this figure does not change from lot to lot as, in the worst case, this could necessitate reformulation to avoid exceeding statutory limits. Consistency of ingredient supply is thus critical.

To calculate the steviol equivalence of a particular stevia extract the proportions of the various components must be known (Table 3). Steviol equivalent concentration is the product of the conversion factor and the concentration of the specific steviol glycoside concerned.


Table 3 Conversion factors



Regulatory* conversion factor to steviol equivalents





Rebaudioside A


Rebaudioside B


Rebaudioside C


Rebaudioside D


Rebaudioside E


Rebaudioside F


Dulcoside A








                                    *(European Commission 2012)

EU regulatory position - labelling

Disclaimer: Acceptable labelling of EU products containing steviol glycosides is not always clear and may differ between member states. The views expressed here have not been tested in the courts and are not to be interpreted as definitive statements of the legal position in any EU member state.


Mandatory labelling applies to the denomination of any product containing steviol glycosides. According to Regulation (EC) 1333/2008 and Directive 2000/13/EC, the product denomination of a food or beverage containing any low-calorie sweetener shall be accompanied by the statement ‘with sweetener(s)’ or if containing both added sugar(s) and sweetener(s) ‘with sugar(s) and sweetener(s)’. For table-top sweeteners the wording is ‘steviol glycoside-based table-top sweetener’.


Also mandatory is the permitted description of steviol glycosides in the ingredient list. As stipulated in Directive 2000/13/EC, this shall be expressed either as ‘sweetener: steviol glycosides’ or ‘sweetener: E 960’.


Voluntary labelling is more open to interpretation. The principal attraction of steviol glycosides is that they are derived from a natural source. This is viewed by many consumers as preferable to synthetic low-calorie sweeteners. Making this natural sourcing clear within a legal label is not always straightforward, however. Although steviol glycosides are made by the Stevia plant and are not chemically altered by any of the extraction and purification steps, the glycosides may not be referred to as ‘natural sweetener(s)’. However, ‘natural’ phrasing may be possible when referring to the source plant material (the stevia leaf).


Accordingly, descriptions such as:-

  • sweetness (sweetener) from a leaf
  • sweet taste from a natural source
  • sweetener (purified /extracted) from a natural source
  • calorie-free sweetener from the Stevia plant
  • Stevia plant (leaf) extract naturally sweet

and others broadly similar, are likely to be acceptable.


It is important to note that pictures used on the label will influence the acceptability. In addition, the use of natural claims should be thought of in the context of both consumer expectation and regulatory authority guidance/enforcement history. In both instances, the considerations may be specific to individual countries.


For example, the FSA publishes guidance on the use of the term ‘natural’ (FSA, 2008), Finland bans the use of the words ‘stevia’ or ‘stevia extract’ where steviol glycosides are involved and gives various examples of both approved and unauthorised expressions relating to stevia (EVIRA, 2013). Belgium likewise lists both banned and allowed descriptions for steviol glycosides, and specifically limits (but does not prohibit) the use of pictures of stevia leaves (Anon, 2012). Other countries may have their own particular regulations on these issues.

Food and drink use of stevia-based sweeteners

As with all high-potency sweeteners, each steviol glycoside’s concentration–sweetness response behaviour is a curve tending to a maximum value of intensity, expressed in % sucrose equivalent (% SE). In other words, the potency of the sweeteners (in terms of the number of times they are as sweet as sucrose) is non-linear and depends on the concentration used. There is no agreed standard concentration, but values around 5% SE are realistic indicators and often used. The potency of reb A in room temperature water at 5% SE is 250 (Fry et al. 2011), but that of stevioside is only half this (DuBois et al. 1991). The latter glycoside is thus at an immediate disadvantage in that twice as much is required to reach the same sweetness intensity as a given concentration of reb A (Figure 2). To compound this disadvantage, regulatory reasons mean the maximum amount of stevioside that can be used in any application is substantially lower than that of reb A. (See conversion factors Table 3 and Figure 4).

In addition, the quality of sweetness available from stevioside is significantly inferior to that from reb A. Both compounds have inherent non-sweet side tastes in addition to sweetness, but reb A is widely regarded as the better-tasting of the two (Phillips 1987; Fry 2012). Control of these side tastes is key to successful product formulation. This encourages formulators to use the lowest practicable concentration of the sweeteners because side tastes are disproportionately less apparent at modest concentrations than at higher levels.

The maximum sweetness available is, however, limited. For example, in soft drinks, legal restrictions will mean that, even using pure reb A, the highest sweetness achievable will be about 5–6%SE (Figure 4). In contrast, most carbonated soft drinks contain around 10% sucrose. Accordingly, steviol glycosides are blended with other sweeteners except where a product, such as a flavoured water, is acceptable with a low sweetness intensity.

While it would be legal to blend with synthetic high-potency sweeteners (e.g. aspartame, sucralose and the like) the preferred candidates for blending are sugars, as these allow a natural ingredient image to be retained. As a result, most stevia-sweetened soft drinks are ‘reduced sugar’ rather than ‘sugar-free’. This development has been encouraged by the fact that the steviol glycosides taste remarkably good in combination with low concentrations of sugars. Figure 3 illustrates the use of reb A in conjunction with different background concentrations of sugar - and some of the EU regulatory constraints on this.

Sucrose is not the only source of sweetness that works well with steviol glycosides, and sugars from fruit, syrups such as honey, agave and fructo oligosaccharides, and sweetness from sugar alcohols all serve too.

While non-alcoholic drinks represent the largest market opportunity for any high-potency sweetener, the steviol glycosides are suited to much wider application. Dairy products, particularly yoghurt, work well and European formulators have already turned their attention to this as well as ice cream and flavoured milks. Sugar-free confectionery is another successful application. Other categories include jams, jellies and fruit spreads. In the USA snack foods, alcoholic drinks and desserts in general also feature stevia sweeteners. Stevia-based table top sweeteners are particularly successful sugar substitutes on both sides of the Atlantic and are presented in powder, tablet or liquid form. They use a carrier to dilute the steviol glycosides so that consumers can readily dispense the sweetness equivalent to that of a teaspoon of sugar. This wide range of products is in part possible because of steviol glycosides’ high stability.

Reb A has been extensively studied and is stable to food processes such as, extrusion, pasteurisation, ultra high temperatures, hot fill and canning. The sweetener is also very stable in baking (Fry 2011a, 2012) and although industrially-baked goods are generally not a permitted application for any high-potency sweetener in the EU, home baking has proved a popular way of using table-top sweeteners in the EU and especially the USA.

Taste Profile

The variability of composition has consequences for the taste of different extracts. As previously noted, the taste of reb A is preferred to that of stevioside. However, these two major glycosides also differ in their effectiveness as sweeteners. Figure 2 contrasts the concentration-response behaviour of stevioside and rebaudioside A. As for all HPS, the concentration-response graph is, in each case, a hyperbolic curve. In contrast to sugars, all HPS become less effective at sweetening the higher their concentration. For this reason potency figures should be accompanied by the sucrose equivalent intensity (SE) at which they were measured. In the case of reb A, its potency in water is 250 at 5% SE (Fry et al. 2011). Figure 2 shows that rebaudioside A is twice as potent as stevioside. Thus, reb A is not only preferred for its taste, but is also a more effective sweetener on a weight basis than stevioside.

All sweeteners differ in their temporal (intensity/time) responses too. Steviol glycosides, in common with other HPS, can exhibit slightly delayed onset of sweetness and some glycosides have a tendency to linger. These qualities vary with glycoside composition and mean each version of stevia has its own challenges when optimising the taste of consumer products.

The steviol glycosides exhibit tastes in addition to sweetness. These non-sweet side tastes include sensations commonly described as bitter and liquorice. These tastes are intrinsic to steviol glycosides and cannot be eliminated by further refining. However, the side tastes of reb A, and mixtures rich in reb A, are generally only apparent at relatively high concentrations and are not significant at lower levels (Prakash et al. 2008). Nevertheless, some flavours (such as citrus and mint) and systems (such as tea-based beverages and alcoholic drinks) are remarkably acceptable with higher concentrations of steviol glycosides. The optimum amount to use is application-dependent and normally located by trial-and-error.

As noted earlier, there is a trend to use leaf extracts containing lower amounts of reb A, blended to maximize beneficial taste synergies between different glycosides. Such blends provide superior sweetness quality to reb A alone or high reb A extracts, especially at higher sweetness intensities. Some suppliers are now developing steviol glycoside mixes particularly suited to specific applications, such as dairy. These improvements are more advanced in the USA, which is free of use limits as well as the EU restriction that steviol glycosides must include a minimum of 75% reb A with stevioside.

Practical Application

A key advantage of the steviol glycosides is that they are seen by consumers as having been derived from nature. Accordingly, despite the limited sweetness intensity allowed by EU regulation, there is some reluctance to blend them with synthetic sweeteners in order to achieve zero or near-zero calorie beverages. The main use of the glycosides is thus in combination with caloric sweeteners in drinks of reduced sugar and/or energy content.

Figure 3 illustrates the effects of such blending with various different concentrations of sugars. The sweetness of the steviol glycosides is additive to that of sugars and the overall sweetness available is thus boosted by the contribution of the background sugar level. This blending permits customary degrees of sweetness to be reached without difficulty. However, in products that have added sugars there is still the regulatory requirement to achieve a minimum 30% calorie saving. This is shown in Figure 3 by dotted portions of the concentration-response curves. It would not be legal to make drinks with the composition shown by the dotted lines, as the calorie saving would be insufficient. In contrast, the solid lines describe fully-legal potential compositions, and it can be seen that not only can the 30% requirement be met readily, but much higher calorie reductions are also attainable within EU regulatory limits.

The success of this approach can be seen in the USA, where a number of major brands enjoy substantial sales of juice-based beverages, partly sweetened with reb A, generally claiming about 50% lower energy contents on their labels, and ranging in juice content from 24% to 75%.

Another trend seen in the USA is the use of erythritol in beverages in combination with steviol glycosides. Erythritol performs exceptionally well in improving the sweetness quality of stevia, and has the advantage of being zero-calorie. Up to 3.5% erythritol is permitted in soft drinks in the USA, although generally lower concentrations are used. Legislative changes in the EU are expected in the last quarter of 2015 to permit erythritol in non-alcoholic drinks at up to 1.6%.


Practical application of the steviol glycosides requires that they are sufficiently stable to provide adequate product shelf life. The glycosides’ stability depends on pH, temperature and time. The effect of these variables is shown in Figure 5, which demonstrates how the time needed for a specified degree of reb A breakdown (25%) to occur depends on temperature for the pH 3-4 range typical of most soft drinks.

The 25% loss figure is a common industry guideline for the maximum acceptable decline in sweetness in those drinks that employ the relatively unstable sweetener, aspartame. However, it should be noted that the breakdown products of reb A are other glycosides, most of which are themselves high-potency sweeteners. This means that a 25% analytical loss of reb A results in only about 7%  drop in sweetness (Fry et al, unpublished), and that Figure 5 gives a highly pessimistic view of the sweetener’s shelf life in terms of taste. In addition, at refrigerated and room temperatures reb A is very stable indeed. Moreover, the sweetener is more than capable of withstanding the heat processing involved in treatments such as pasteurisation and hot fill as well as UHT and canning.


Steviol glycosides are non-caloric, high-potency sweeteners extracted from the leaves of Stevia rebaudiana. The glycosides, permitted as sweeteners in the EU and widely elsewhere, are ideal for use alone or blending with caloric sweeteners in the formulation of a range of reduced sugar foods. The glycosides are stable to processing and offer few shelf life limitations.


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