Food and Chemical Toxicology 107 (2017) 302e317

Contents lists available at ScienceDirect

Food and Chemical Toxicology

journal homepage: www.elsevier.com/locate/foodchemtox

Review

Sweeteners as food additives in the XXI century: A review of what is

known, and what is to come

rcio Carocho a, Patricia Morales b, *, Isabel C.F.R. Ferreira a, **
Ma
a
nia, 5300-253, Bragança, Portugal
Mountain Research Centre, (CIMO), ESA, Polytechnic Institute of Bragança, Campus de Santa Apolo
b
n y Cajal, s/n, 28040, Madrid, Spain
Department of Nutrition and Bromatology II, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramo

a r t i c l e i n f o a b s t r a c t

Article history: Sweet has always been a very important basic taste for mankind, although sweetness is always related to
Received 23 May 2017 either weight gain or teeth decay. Sweeteners entered the food industry back in the 1800's and are now
Received in revised form staple in foodstuffs. Despite their long relationship with food, sweeteners have been in the spotlights for
26 June 2017
many reasons. Since being the perfect choice for diabetics, to the dangers concerning toxicity, cancer and
Accepted 28 June 2017
Available online 5 July 2017
other health issues associated with their consumption, sweeteners have come a long way. The conflicting
results for the same sweeteners and the divergent regulations are fuel for a wide debate on the impact of
sweeteners in the industry, health and lifestyle of mankind. In this review, the history, main concerns,
Keywords:
Food additives
benefits, disadvantages, classification and future trends are revisited for nutritive, intense and natural
Sweeteners food additives, while future perspectives are hypothesized.
Natural sweeteners © 2017 Elsevier Ltd. All rights reserved.
Intense
Nutritive

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 302
2. General considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
3. Classification, properties and sweeteners role in food . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 303
3.1. Polyols: classification and applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305
4. Intensive sweeteners . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
4.1. Synthetic intensive sweeteners . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
4.2. Natural intensive sweeteners . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311
5. Conclusions and future perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 314
Conflict of interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315
Transparency document . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315

1. Introduction human condition makes humans tend to prefer some stimuli, and
hate other types of negative sensations (Mooradian et al., 2017).
Mankind experiences the world through its five primary senses, With regard to taste, it is widely known that children prefer sweet
and interprets the inputs with reason and emotion. This fragile taste over the other basic flavors, and although this can change with
age, sweet taste is still one of the most desired flavors for mankind,
preferred over sour and bitterness (Kim et al., 2017). Research
* Corresponding author. deems this preference as innate and linked it to sensations of
** Corresponding author. pleasure and happiness. Moreover, other studies have proven that
E-mail addresses: patricia.morales@farm.ucm.es (P. Morales), iferreira@ipb.pt
sugar craving could be genetic, making some individuals struggle
(I.C.F.R. Ferreira).

http://dx.doi.org/10.1016/j.fct.2017.06.046
0278-6915/© 2017 Elsevier Ltd. All rights reserved.
 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317 303

with adverse effects of overconsumption of sweet food (Drexler and Table 1

Sou cek, 2016; Padulo et al., 2017). Difference of sweetness among different molecules, calculated
based on the assumption that sucrose is equivalent to 1 unit of
In a profit driven economy, the food industry finds new ways to sweetness. Data extracted from Mitchell (2006); Otabe et al.
lure consumers to consume their products, paying little attention to (2011c); Varzakas et al. (2012).
the potential health effects of the cumulative consumption of
Sweetener Sweetening power
various food products throughout the day. Thus, especial attention
should be given to children and toddlers, for they are unaware of Advantame 37000
Neotame 7000e13000
the consequences (Hert et al., 2014). For many years, the con-
Neohesperidin 1500e2000
sumption of excessive sugar is known to have adverse effects in Sucralose 400e800
humans, and thus, to reduce its intake, sweeteners appeared back Saccharin 240e300
in the 1800's. Since their inception, sweeteners have come a long Aspartame 200
Acesulfame-K 150e200
way, and while they were once regarded as one of the most
Cyclamate 30e80
important achievements for the food industry, many controversies, Fructose 1.1e1.5
conflicting regulations and laws have deemed them to untrusted Sucrose 1
molecules that are added to food to make it sweeter. Today, Xylitol 1
metabolic disorders are increasing, obesity is increasing, and so are Dextrose 0.9
Maltitol 0.75
diabetes and other diseases that are directly related to sugar con-
Glucose 0.75
sumption, in fact, the WHO studies have considered metabolic Erythritol 0.7
disorders as being of “epidemic proportions” in industrialized Mannitol 0.6
countries (Pradhan, 2007; Rani et al., 2016). With the increasing Sorbitol 0.6
prevalence of diseases related to sugar consumption, sweeteners Isomaltose 0.55
Maltose 0.4
are now widespread in foodstuffs, and are highly researched for Lactitol 0.35
their impact on sweetening potentials, on health, on the economy Galactose 0.3
and in social studies (Mooradian et al., 2017). Raffinose 0.2
In this article, the authors review the sweeteners state of the art
(classes, behaviors, applications, benefits, disadvantages, scandals,
and corporative tactics), and postulate future trends in food 1971). For a compound to display a sweet taste, the molecular
sweetening. distance between A and B must be at least 0.25e0.40 nm. The
sensation of sweetness also depends on the configuration of the
2. General considerations molecule, which in sugars comes from the dextrorotary confor-
mations, but not from the inverse, levorotary. For this reason, some
The most important aspect of sweeteners is undoubtedly their sugars, like cellobiose are insipid, but others are sweet (D-glucose),
sweetness; it is measured in relation to sucrose, which is the while L-glucose is slightly salty. The three-dimensional structure of
reference sugar. Thus, a solution of 30 g L
1 at 20  C has a sweet- D-glucose binds to the receptor with the hydroxyl group of C4 (AH
ening power of 1, with the threshold of the minimum concentration function) and the oxygen in C3 (B function), together with the
to detect sugar being 1e4 mM. For the sweet flavor intensity to be hydroxyl group of C6, they bind to the receptor through hydrogen
perceived, the substance must first be dissolved in saliva and come bonds (Crammer and Ikan, 1979; Lindemann, 2001). The activation
into contact with the receptors that are present on the tongue. of such receptors by sweet substances releases adenosine triphos-
Other parameters influence the sweet flavor, namely the sugar phate (ATP) that consequently activates neurons that transmit this
structure (in which the intensity decreases as the number of input to the brain. Apart from depending on physical and chemical
monosaccharides increase), temperature of perception, pH and the properties of the substances, sweet flavor is also determined by
presence of other molecules that can influence receptors. differences in humans, namely age, genetics, race and ethnicity
Taking sucrose as a reference (its reference value can be (Nelson et al., 2001; Ohtsu et al., 2014; Smutzer et al., 2014;
considered 1), it has a higher sweetening power when compared to Weerasinghe and DuBois, 2006).
other simple sugars, for instance, galactose (0.3), while others show
higher values, namely fructose (1.7). Furthermore, there are other
highly complex sweeteners that have thousands of times the 3. Classification, properties and sweeteners role in food
sweetening power of sucrose, neotame (13,000 times sweeter) or
even advantame, which is 37,000 times stronger than sucrose Sweeteners (as legal food additives and non-additives) can be
(Table 1). classified by intrinsic properties or origin. Some of the most com-
Biologically, the perception of sweetness happens through the mon classifications are in terms of their nutritive value, sweetening
receptors on the taste buds, based on a donor/acceptor proton power and their provenance. Thus, they can be divided in nutritive
system, establishing an AH/B/X system between the food and the vs. intensive sweeteners (Fig. 1A and B), but also between synthetic
receptors of the taste buds. A and B are electronegative atoms, like and natural origin (Fig. 2). While the former is a classification used
oxygen or nitrogen, H represents hydrogen, which is connected to by governing bodies like the European Food Safety Authority of the
an atom (A) through a covalent bond. X represent hydrophobic European Union (EFSA) the division in natural and synthetic food
groups that are attracted by the taste buds in order for the AH/B/X additives is not used by these organizations, and is based solely on
to become tridimensional. The receptors of the taste buds are the origin of the sweetener (see Fig. 3).
coupled to G proteins (T1R2 and T1R3), forming part of the C class In the nutritive sweeteners group (Fig. 1A) are the simple sugars
of proteins (GPCR), which are structurally similar to the glutamate but also high fructose corn syrup, isomaltulose, trehalose, which,
metabotropic receptors. The bond between sweet molecules with under the Regulation (EU) No 1333/2008 cannot be considered food
the AH/B/X structure with these receptors happens through hy- additives, but ingredients. Furthermore, polyols (which are
drophobic hydrogen bonds. This bond changes the configuration of considered as food additives) are also included in this classification;
the “taste sensible” receptor, altering the permeability of the ionic examples of polyls are erythritol, isomaltitol, lactitol, maltitol,
environment, aiding the entrance of Naþ (Shallenberger and Acree, sorbitol, mannitol, and xylitol. On the other hand, the intensive
304 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
Fig. 1. A e Examples of nutritive sweeteners. B e Examples of intensive sweeteners.
sweeteners, all of them considered as food additives (Fig. 1B), have
negligible caloric contribution and high sweetening capacity, being
used in low quantities in food. Generally, they are not cariogenic
and do not trigger glycemic response, thus being extensively used
in hypocaloric diets, for diabetes patients and other specific cases
where caloric intake must be controlled (Baines and Seal, 2012;
Varzakas et al., 2012).
The other classification, presented in Fig. 2 divides sweeteners
by their nature or origin, which can be either synthetic or natural.
Sucrose, a disaccharide, the most used table sugar, known
commonly as sugar and the most used sweetening agent in the
world. It is composed of a molecule of glucose in which the alde-
hyde carbon is joined to the ketone one of fructose, forming a b(1,2)
bond, preventing any reducing properties and forming an adequate
structure to bind to the taste bud receptors, conferring the tradi-
tional sweet taste. For some time now, the relation between the
consumption of this sugar and dental decay has been established,
given that it is the substrate for bacteria, like Streptococcus mutans
and S. sanguis that use this disaccharide and convert it to pyruvic,
acetic and lactic acid, which dissolve the tooth enamel, fostering
bacterial colonization. Furthermore, the very fast absorbance of
sucrose can make glycemic values spike, causing hormonal prob-
lems, thus the danger if its consumption by some diabetic patients.
Other diseases and disorders are also related to sugar consumption,
among them are cardiovascular diseases (coronary), type II dia-
betes, metabolic syndrome, hypertriglyceridemia, insulin resis-
tance, cancer (breast, colon), obesity, childhood obesity,
hypertension and kidney diseases (Bostick et al., 1994; Grundy,
1999; Johnson et al., 2007; Ludwig et al., 2001; Mente et al.,
2009; Slattery et al., 1997; Stanhope et al., 2013; Touger-Decker
and van Loveren, 2003; Yang et al., 2014).
For these reasons, the role of sweeteners has been paramount in
the dichotomy of food and health, and their impact on our daily life,
longevity and quality thereof is staggering. Thus, the changes of
food consumption have been drastic, with sweeteners chugging
alongside the industrialization of food and food components, being
their discovery a revolution in the food sector. This allowed the
production of sweet food without caloric intake, changing the
paradigm of how we eat, and of vital importance to people with
hypocaloric diets and diabetics. For the latter, polyols, for instance,
as like fructose, display a metabolization that is independent from
insulin, given that they may enter the hepatic cells through the
 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317 305

Fig. 2. Examples of synthetic and natural sweeteners.

A B

Fig. 3. Chemical structure or sorbitol (A) and mannitol (B).

action of the enzyme fructocinase, which is independent from in-
sulin, which also deems them safe for these patients. Furthermore,
many sweeteners are not cariogenic, so they are not used as sub-
tract for oral bacteria.
Some of the most used sweeteners worldwide are aspartame
(E951), cyclamates (E952), acesulfame K (E950), tagatose (consid-
ered “generally regarded as safe” (GRAS) by the Food and Drug
Administration of the United States of America (FDA)), sucralose
(E955) and more recently steviol glucosides (E960).
3.1. Polyols: classification and applications
Polyols, polyhydric alcohols or polyalcohols, are food additives
that result of the hydrogenation of reducing sugars, being the
presence of an alcohol group in the place of the carbonyl group
quite common in the aldose or ketose fractions of mono, di, oligo
and polysaccharides. Polyols are stable at high temperature, pH
changes and do not intervene in Maillard reactions. They can be
found in nature, especially in fruit and vegetables, being partially
responsible for their sweetness. Their industrial production started
in the 20's with hopes of solving health problems related to
excessive sucrose consumption. Nearly 90 years after, polyols are
the most consumed group of sweeteners because of their lack of
cariogenic properties, salivation induction, and no interference in
insulin levels, being used in “light” foods (Nabors, 2001 (Chapter II);
Varzakas et al., 2012). On the other hand, their consumption is not
recommended for toddlers under 1 year of age, due to their laxative
effects that can unleash severe diarrheas. Technologically, polyols
are also important, namely for reduction of water activity, as
humectants, inert to Maillard reactions, texturizing agents, sugar
crystallization mediators, flavoring solubilizers, and so on. When
being used for these purposes, they are labelled quantum satis (latin
for “just far enough” meaning that there is no limit for its use, as
long as the least amount for the specific outcome is used) (EU
Regulation 1333/2008; EU Regulation 1129/2011). The most used
polyols are sorbitol (E420), mannitol (E421), isomaltose (E953),
maltitol (E965), lactitol (E966), xylitol (E967), erythritol (968). Less
used, but also with relevance are arabitol and hydrogenated starch
hydrolysates (HSH), although not allowed within the EU.
Sorbitols (E420) and mannitols (E421) are isomeric polyols and
have been use in food since the 40's through glucose syrups,
inverted sugars and other hydrolyzed starches. Their production is
based on the catalytic hydrogenation of glucose with subsequent
purification. The separation of the isomers is done by solubility
difference, yielding a very hygroscopic sorbitol and a much less
mannitol. Sorbitol is 50e60% sweeter than mannitol, making it
preferably used as sweetener, while mannitol is employed as an
anti-caking agent (Song and Vieille, 2009). Overall, these polyols
are used in baked goods, sweets, bubble gum, surimi, sausages and
drinks (Nabors, 2001 (Part II)). Although there is no evidence of
sorbitol toxicity, in 2016, a study found that sorbitol can be geno-
toxic and induce metabolic reactions in offspring of female wistar
albino rats fed sorbitol. A study found that patients with irritable
bowel syndrome (IBS) have adverse gastrointestinal reactions to
polyols, especially sorbitol and mannitol, being these reactions in-
dependent of the absorption patterns of each molecule. While
sorbitol can be of concern for patients with IBS, it seems to be safe
for healthy individuals, although there are reports of laxative
306 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317

effects when consumed in high doses. Some studies point out that
this effect is related to the fructose:glucose:sorbitol ratio that is
consumed, and not to sorbitol itself (Hoekstra et al., 1993; Yao et al.,
2014). Mannitol, although less sweet than sorbitol is also used in
food, given its high metabolization ratio, about 75%, being the other
25% absorbed before being excreted in urine. Because it is virtually
inert, it does not react with active components of drugs and confers
a cool sweet taste, apart from being used in the food industry, it is
also widely used in the pharmaceutical area in dental hygiene
products, drug filler and as a diuretic in intravenous fluids
(Biesiekierski et al., 2011; Lee, 2015; Livesey, 2003; Mitchell, 2006).
Isomaltose, isomaltitol, or isomalt (E953) in a legal polyol in the
EU and the USA, obtained through enzymatic transformation of
sucrose (Cammenga and Zielasko, 1996). It is stable at high tem-
peratures and has a very low hygroscopic value. Its sweetening
power is in line with other polyols, about 45e60% of sucrose, but a
very low caloric contribution, of about 2 kcal g
1. This molecule is
Fig. 5. Chemical structure of lactitol.
not absorbed by the small intestine, and is readily fermented in the
colon through colonic bacteria (Caballero et al., 2016). Isomalt is
used in bubble gums, gelatins, chocolate, coatings, baked goods and hydrogenation of starch, resulting in a sweetener with about 90% of
yogurts, among others (Grenby, 1996; JECFA, 2003). (See Fig. 4). sweetening capacity, no other residual flavors and very high sta-
Lactitol (E966) is a disaccharide that is obtained by hydroge- bility (Maguire et al., 2000; Pratt et al., 2011). Of all the sugar al-
nation of lactose. It was discovered around 1920 and since then has cohols, it is the one that most resembles the flavor of sugar. It is not
been used in many different foods. Given its limited sweetening cariogenic and safe for diabetics. In terms of solubility and hygro-
power compared to the other polyols it is usually used in combi- scopicity it is very similar to sucrose, thus being the preferred sugar
nation with intense sweeteners, like acesulfame K, aspartame and to use in the production of chocolate in which the label says “no
sucralose. The human body does not metabolize this polyol, sugars added” (Joshi et al., 2016). It has a very slow digestion rate,
therefore it has no caloric contribution (Mitchell, 2006). Still, it only being fermented in the colon. Apart from chocolates, it is also
has a sweetening power of 30e40% of sucrose and a lower solu- employed in lactic products, baked goods, muffins, bubble gum,
bility that xylitol and sorbitol. Its taste, apart from being sweet gives jams, jelly and other sweets (Grenby, 1996; Mitchell, 2006). (See
a fresh aftertaste, and is therefore used to confer different kind of Fig. 6).
sweetness to food. Furthermore, it is also used to increase food Xylitol (E967), a five-carbon polyol, obtained by hydrogenation
volume, as a probiotic, while also not being cariogenic. Lactitol of xylose, was first synthesized in 1891 and has about 95% the
exists in four crystallized crystalline forms: anhydrous lactitol, sweetness of sucrose. Of all the polyols, it is the sweetest,
lactitol monohydrate, lactitol dehydrate, and lactitol trihydrate, contributing only 2.4 kcal g
1. This compound is obtained by
being the anhydrous the most stable form of this compound. The extraction from birch and other woods, almond husks, corncobs,
foods it is used in are chocolates, baked goods, bubble gums and ice straw and paper production surplus. Apart from this, and although
creams (Aidoo et al., 2013; Grenby, 1996; Halttunen et al., 2001; not viable for industrial production, xylitol is also found in many
Mitchell, 2006). (See Fig. 5). fruits and vegetables (Nabors, 2001 (Part II); Mitchell, 2006). It is
Maltitol (E965) is obtained by hydrolyzation, reduction and known to increase salivation, thus increasing teeth cleansing and
reducing the bacterial load in the mouth and therefore teeth decay.
Xylitol is used in bubble gum, refreshments, baked goods, among
others. It is estimated that this sweetener has a market of 670
million dollars worldwide, which has been increasing 6% per year,
and is expected to continue growing until 2020. These figures, in
line with what is happening transversally to all food additives,

Fig. 4. Chemical structure of isomaltose. Fig. 6. Chemical structure of maltitol.

 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
represent the increasing awareness of naturally derived food ad-
ditives (Dasgupta et al., 2017; Mitchell, 2006; Peterson, 2013). (See
Fig. 7).
Erythritol (E968), used legally both in the EU and the USA, ap-
pears naturally in some fruits (melon, pears and grapes), but also in
vegetables, mushrooms, honey and seaweeds, but its primary
method for industrial production is through yeasts (Boesten et al.,
2015 Tomaszewska et al., 2014). Being discovered in 1848, today
it takes part in a multitude of products, like food coatings, baked
goods, fermented milk, glazed goods, candy, and chocolate
(Carocho et al., 2015). Its sweetening power is about 60e80% of that
of sucrose and a very low caloric contribution, of just 0.3 kcal g
1,
therefore being safe for diabetics. Furthermore, erythritol is easily
absorbed through the intestine and has a very low metabolization,
being almost completely excreted in urine (Arrigoni et al., 2005;
Ro€ per et al., 1993). Erythritol is considered as a safe additive after
many specific tests regarding toxicity, carcinogenicity and repro-
ductive hazards resulted negative, although in 2013, there was
report of an 11-year-old child that had erythritol induced anaphy-
laxis (Shirao et al., 2013). This sweetener also displays antioxidant
capacity and has protective endothelial properties (Boesten et al.,
2015). (See Fig. 8).
Apart from these polyols, there are others, although these
cannot be used in food inside the EU, namely arabitol, which is
obtained by reduction of arabinose, and HSH, which is a mixture of
polyols that can achieve about 90% of sweetening power. In the
USA, these sweeteners are legal, and actually considered GRAS food
additives. Arabitol has a very similar structure to sorbitol (6-carbon
skeleton) and is used for its rheological properties, viscosity
improvement, humidifying, crystallization and rehydration prop-
erties of the foodstuff it is used in (Carpentier et al., 2013;
Modderman, 1993). HSH are a family of bulk nutritive sweeteners
that comprehend hydrogenated glucose, maltitol and sorbitol
syrups. They were first developed in the 60's in Sweden and have
been used in foodstuffs since then. They are produced by hydrolysis
and hydrogenation of corn, wheat or potato starch, and a caloric
contribution of 3 calories per gram, not causing tooth decay.
Furthermore, HSH can be used as viscosity and bodying agents,
humectants, crystallization modifiers and rehydration aids. (Larry
and Greenly, 2003). On Table 2 some of the most important poly-
ols used in food are displayed, along with their chemical structure
and caloric contribution per gram. (See Figs. 9 and 10).
4. Intensive sweeteners
Intensive sweeteners are those that present a high sweetening
power, higher than sucrose, thus only being necessary in very low
doses to obtain intense sweetness. Their caloric contribution is also
very low or even virtually zero, they also present no danger in terms
of cariogenicity or insulin reaction and have no other function in
food apart from sweetening.
4.1. Synthetic intensive sweeteners
Among all the intense sweeteners used in the industry, the most
Fig. 7. Chemical structure of xylitol.
307
Fig. 8. Chemical structure of erythritol.
Table 2
Most used polyols and caloric contribution in kcal g
1. Data extracted from Varzakas
et al. (2012).
Name Caloric contribution
Hydrogenated starch hydrolysates 2.8
Maltitol - E 965 2.7
Sorbitol - E 420 2.5
Xylitol - E 967 2.5
Isomaltose - E 953 2.1
Lactitol - E 966 2
Mannitol - E 421 1.5
Erythritol - E 968 0.2
notable ones are acesulfame K (E950), aspartame (E951), cycla-
mates (E952), saccharin (E954), sucralose (E955) and neotame
(E961), which are detailed in Table 3. Recent research has showed
the impact of neural mechanisms involved in the sweet taste, and
related that, like all things sweet, sweeteners modulate neural
systems, perpetuating their intake, although in different manners.
Both caloric and non-caloric sweeteners act on the reward mech-
anisms, and in situations of caloric deficit, caloric sweeteners
excerpt stronger craving. Thus, non-caloric sweeteners potentially
act with lower intensity in neuronal paths of reward, but is their
contribution to health and industry innocuous? Probably not …
Recent studies suggest that non-nutritive sweeteners can, sur-
prisingly, be related to weight gain and type 2 diabetes risk through
3 potential mechanisms: a) interference with learned responses
that contribute to control glucose and energy homeostasis; b)
interference with the gut microbiota, inducing glucose intolerance;
c) interaction with sweet-taste receptors that may trigger insulin
secretion (Murray et al., 2016; Pepino, 2015).
Acesulfame K (E950) corresponds to the potassium salt of ace-
sulfame, and was discovered in 1967, although today, its industrial
production has changed, being obtained through sulfamic acid and
deketene which will eventually produce sulphur. Acesulfame K is
one of the most used synthetic sweeteners due to the lack of re-
sidual flavors and a sweetening power of over 200 times the one of
sucrose. It can be used in synergies with other sweeteners, namely
aspartame, cyclamates and sucralose to further improve sweet-
ening and flavor. Unlike polyols, this compound suffers metabo-
lization by the human body, thus having an admissible daily intake
(ADI) of 15 mg kg
1 of body weight. The ADI is the maximum
amount of a compound that can be ingested per kg of body weight
per day, considering all the sources of the compound. Many studies
have described its innocuousness, although other studies up until
2000 pointed some type of toxicity, but were then disproved
(Carocho et al., 2014; Shankar et al., 2013). Still, studies carried out
by Stohs and Miller (2014) claim that there is some type of hyper-
sensitivity at a dose dependent manner. Acesulfame K is used in
baked goods, cereals, sweets, confectionary products, marmalades,
canned food and fruit, bubble gum and as tabletop sweeteners (for
coffee and others) (Nabors (Part I), 2001; Mitchell, 2006; O'Donnell
and Kerasley, 2012). A new problem related to acesulfame k and
other synthetic sweeteners is their ubiquity in the environment
given the high amounts that are consumed by populations and
308 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317

A B

Fig. 9. Chemical structure of arabinose (A), the precursor of arabitol (B).

A B

Fig. 11. Chemical structure of sulfamic acid (A) and acesulfame K (B).

Fig. 10. Chemical structure of hydrogenated starch hydrolysates. prolonged heat and pasteurization procedures, thus having very
high stability, even higher then saccharin and acesulfame K. It has a
pleasant taste without sourness or metallic residue (usual in some
Table 3
types of sweeteners) and a sweetening capacity of 180e200 times
Structure, ADI and sweetening power of the most common synthetic intense
sweeteners. Data extracted from Mitchell (2006); Otabe et al. (2011c); Varzakas et al. sucrose. It is however, considered a source of phenylalanine thus
(2012). not being advised for people with phenylketonuria (Shankar et al.,
2013). Furthermore, there are reports of toxicity and hepatocellular
Name ADI (mg/kg/day) Brand Names Sweetening power
alteration in long-term exposure to it (Abhilash et al., 2011). Ac-
Advantame - E 969 5 e 37000
cording to regulation of the EU No. 1169/2011, all food that uses
Neotame - E 961 2 Newtame® 7000e13000
Sucralose e E955 5 Splenda® 400e800 aspartame has to have a visible section containing “contains
Saccharin - E 954 5 Sweet and Low® 240e300 aspartame (source of phenylalanine)”. Its ADI is 40 mg kg
1 of body
Sweet Twin® weight. Further recommendations should be considered, given that
Sweet N'Low® the use of aspartame in food with a pH higher than 6 can make it
Aspartame - E 951 40 Nutrasweet® 200
Equal®
transform into diketopiperazine, a carcinogenic compound (Rycerz
Sugar Twin® and Jaworska-Adamu, 2013). Aspartame is used in refreshments,
Acesulfame K - E950 9 Sweet One® 150e200 yogurts, lactic beverages, desserts, baked goods and others
Sunett® (Mitchell, 2006; Nabors (Part I), 2001; O'Donnell and Kearsley,
Cyclamates - E 952 11 Assugrin® 30e80
2012). It has always been a target of intense studies regarding its
Chuker®
Cologran® safety, although being widely regarded as safe, this sweetener has
Hermesetas® seen recent studies, (2010 and onwards) pointing outs its nephro-
Huxol® toxicity, hepatotoxicity, damage to nerves, cancer, and even type 2
Novasweet® diabetes (which is strange given the non-nutritional nature), being
Rio®
all these diseases reported in murine models (Ashok et al., 2013;
Sucaryl®
Sugar Twin® Fagherazzi et al., 2013; El Haliem and Mohamed, 2011; Okasha,
Suitli® 2016; Saleh, 2014; Soffritti et al., 2010). Despite having evident
Sweet N'Low® differences, the human body and metabolism shares some simi-
larities with these models, which perpetuates the mistrust of this
molecule by the public. Inversely, a review published in the Food
excreted into wastewaters. Thus, a considerable amount of research and Chemical Toxicology Journal, authored by Kirkland and Gate-
groups are trying to find new ways to inactivate this contaminant, house in 2015, deems aspartame safe, after reviewing all available
for it is excreted unaltered due to the lack of metabolization in the data from various sources. The authors state no toxicity in gene
human body. In surface waters, its concentration can achieve mutations, some evidence of chromosomal damage in vitro, while
1 mg L
1, which is higher than the concentration of the average bone marrow micronucleus, chromosomal aberration and comet
contaminant. The major problem is that the residue produced by its assays found no toxicity in somatic cells, supporting the claim by
inactivation is more toxic than acesulfame k itself, which consists of the EFSA of a non-genotoxic compound. In 2014, an article signed
yet another challenge to the food/environmental industries with by Suez et al. and published in Nature referred to aspartame as
regard to synthetic food additives (Yin et al., 2017). (See Fig. 11). inducing glucose intolerance by altering gut microbiota, requesting
Aspartame (E951), discovered in 1965 is obtained through the that a reassessment of non-caloric artificial sweeteners be carried
combination of amino acids, namely L-phenylalanine, L-aspartic out. Concomitantly, PepsiCo announced in 2015 that they would
acid, and connected through methyl ester bonds. Given is remove aspartame from their diet version of the famous drink,
extremely low solubility in water and unstable pH it cannot be used bowing to consumer demand of an aspartame free drink. In 2016,
in refreshments with low pH like juices, and does not resist to after a crash in sales, PepsiCo reintroduced aspartame in their
 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
beverage, claiming that some consumers found a difference in
taste, making the company produce three types of beverage: the
original Pepsi, one with aspartame and one with a natural sweet-
ener to meet all customer demands. This small industrial maneuver
further deepens the plot and casts more distrust and fear in cus-
tomers, which are left baffled without actually knowing if aspar-
tame is safe or not (CNBC, 2015, 2016; Paolini et al., 2016). (See
Fig. 12).
Cyclamates (E952), discovered in 1937 at the University of Illi-
nois, are a very good example of the legislative discrepancies be-
tween the EU and USA. The European Union has approved its use in
food, while the FDA removed its GRAS status in 1969 and
completely banned it in 1970. It is now pending approval for re-
admission. The base for this ban is a study that relates the metab-
olization of cyclamates to cyclohexylamine (toxic compound), and
although a later study pointed out that this metabolization only
takes place in a small amount of the population, it has not been
enough for the FDA to remove the ban. Its industrial production
could be the reason for this intransigent maneuver, given that it is
obtained though the sulfonation of cyclohexylamine (Renwick and
Nordmann, 2007). China, Indonesia, Taiwan and Spain are the
biggest producers of this sweetener, which, along with saccharin
are the least expensive to produce (Nabors (Part I), 2001). In the EU,
the ADI is set at 11 mg Kg
1 of body weight and is used in desserts,
baked and processed food, soft drinks, canned fruits, gelatins and as
tabletop sweeteners (Carocho et al., 2014). One of the drawbacks of
cylamates is the slight sour taste, although its sweetening capacity
is set between 35 and 50 times stronger than sucrose. The long-
lasting sweetness if put off by a sour aftertaste, so it is almost al-
ways combined with saccharin (Martins et al., 2010; Mitchell, 2006;
O'Donnel and Kearsley, 2012; Renwick et al., 2004; Roberts, 2016;
Takayama et al., 2000). (See Fig. 13).
Saccharin (E954) was the first discovered intense sweetener,
back in 1878. Today, more than 100 years later, it is produced at an
industrial scale, though a process called Maumee, which derives
from the company that developed the technique (Maumee Chem-
ical Company). In this process, phathalic anhydride is converted to
anthranilic acid to then react with nitrous oxide, nitrogen dioxide,
chlorine and ammonia, forming saccharin Carocho et al., 2014
Mitchell, 2006. This compound is stable at low pH and resists
high temperatures, which makes it an ideal sweetener to be used in
production steps of foods and for long lasting goods. It has a sweet
taste, but also a slight acid contamination so it is combined with
A B
C nations, water consumption, clinical pathology evaluations, no re-
Fig. 12. Chemical structure of the amino acids L-phenylalanine (A), L-aspartic acid (B),
which are the building blocks of aspartame (C).
309
B
A
Fig. 13. Chemical structures of the precursor cyclohexylamine (A) and sodium cycla-
mate (cyclamate) (B).
cyclamates and aspartame. It can have 300 times the potency of
sucrose in terms of sweetening, but has the lowest ADI of all non-
caloric sweeteners, only 5 mg Kg
1 of body weight. In terms of
consumption, it has always been controversial, with Canada ban-
ning its use in 1977 after animal testing showed toxicity. In that
same year, the US considered doing the same, but Congress placed a
moratorium on this decision, having other studies then ruled out
the adverse effects. All these studies were based on the formation of
tumors in rats, namely in the bladder (Nabors (Part I), 2001). Thus,
given the anatomical differences between mouse and man, the
danger for humans was ruled out. Today, numerous studies have
deemed saccharin safe and its consumption is, today, safe, fostering
the increase of its use all over the world (Shankar et al., 2013). As
like acesulfame K, saccharin is excreted through urine and is not
metabolized in the body, although it can cross the placenta of
pregnant woman and can be transferred through breast milk, thus
not being recommended to pregnant women or breastfeeding ones.
It is employed in fruit juices, processed fruit, gelatins, marmalades,
coverings, sauces, desserts, bubble gum and tabletop sweetener
(Swithers, 2013; Takayama et al., 1998; Whysner and Williams,
1996).
Neotame (E961) is a sweetener with a very similar structure to
aspartame, in fact, they are isomers, but neotame has a very high
sweetening power, from 7000 to 13000 times stronger than sucrose
and less than 1.2 kJ g
1 Mayhew et al., 2003. It has a clean taste,
with no metallic or acid aftertaste. Like sucralose, it was only
discovered in the 80's and is obtained by reductive alkylation of
aspartame, which is converted into 3,3-dimethylbutyraldehyde.
Given the lack of phenylalanine in its composition, it is safe for
phenylketonuria patients, but also safe for diabetics O'Donnell and
Kearsley, 2012. It is used mainly in synergies with other sweeteners
(except acesulfame-k and saccharin) and for soft and lactic drinks,
sauces, yogurts, lemon tea, as tabletop and in bubble gum, but also
as an enhancer of natural flavors, namely acidic fruit flavors. It is
stable under dry storage conditions, not hygroscopic and appears a
white crystalline odorless powder. In terms of metabolization, half
of the ingested neotame is not absorbed and excreted through the
feces while the other half in excreted in the urine as de-esterified
neotame. It meets the five basic criteria for commercial viability
of a nonnutritive sweetener: taste, solubility, stability, safety and
cost. Regarding safety, neotame, as has been subject to a battery of
tests, in which, even at doses higher that its ADI, no toxicity was
detected. No adverse findings were reported for physical exami-
ports of morbidity, mortality, organ toxicity, macroscopic or
microscopic postmortem findings in murine models and other test
animals (Mitchell, 2006; Nabors (Part I), 2001; Nofre and Tinti,
2000; Zhu et al., 2016).
Advantame (E969) is one of the newest sweeteners to be
approved in the EU, in 2013. It is obtained through chemical syn-
thesis from aspartame and isovaniline. Contrary to neotame,
advantame is a source of phenylalanine, and even though it is
310 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
derived from aspartame, it has a very different structure. The
sweetening power of this molecule is around 20,000 times the one
of sucrose and it appears a white to yellow powder (Warrington
et al., 2011). It has a very sweet flavor with little intensity of
bitter and sour. It has been successfully used in coffee, iced tea,
powdered beverage formulations, chewing gum, yogurt, and also as
a flavor enhancer (Otabe et al., 2011a). In terms of stability, it can
withstand high temperatures and be used in low pH products. In
July 2013, EFSA experts defined advantame as non-toxic or carci-
nogenic and there are no risks of its consumption as a food additive.
The ADI was established at 5 mg kg
1 of body weight per day.
Model animal (rats, dogs) and human trial data suggest that there
are no issues with the use of advantame as a food sweetener (EFSA,
2013; Otabe et al., 2011b, 2011c; 2011d; Ubukata et al., 2011). (See
Fig. 14).
Sucralose (E955) is another intense synthetic sweetener, ob-
tained industrially by substitution of the three hydroxyl groups in
sucrose Roberts et al., 2008. This transformation renders this
molecule 750 times sweeter than the precursor, sucrose. Its ADI is
Fig. 14. Chemical formulas of A e saccharine, B e neotame, and C e advantame.
5 mg kg
1 of body weight (as saccharin), although sucralose suffers
metabolization in the human body. This metabolization is related to
migraines, intestinal unrest and inhibition of colonic bacteria when
consumed in high amounts. The principal applications of sucralose
are yogurts, ice cream, canned fruits, biscuits, caramels, soft drinks,
lactic products, baked goods, gelatins, marmalades, bubble gum,
among others Mitchell, 2006; Nabors (Part I), 2001; O'Donnell and
Kearsley. Despite some research pointing out the possible
connection of the consumption of sucralose with cancer at the
beginning of the century, a review article authored by Berry et al.
(2016), stressed that there is no relation, even at higher dosages
that the previewed in the ADI. Furthermore, back in 2009, another
review article that thoroughly reviewed all the available studies at
the time pointed out that there are no risks in consuming this
additive, and many studies in vivo, in vitro and with human trials
point out the overall safety of sucralose Grotz and Munro, 2009; In
2012, another paper refuted claims of previous ones that ques-
tioned the safety of sucralose, namely in stability in vivo, chemical
reactivity of sucralose, stability at high temperatures and
 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
interaction with cytochrome P450 Shiffman and Abou-Donia, 2012.
Recently, more controversy shattered the public opinion, when in
2016 an article pointed out that consumption of sucralose increases
food intake through a neuronal fasting response Wang et al., 2016.
The research was carried out in Drosophila flies for their simple
genome and fast replicating speeds. Recently, in the same journal,
Cell Metabolism, a new research implies the contrary, stating that
sucralose suppresses food intake, using the same test subjects.
Another study, dated 2017 claimed that children, due to their lower
weight and blood volume, have a higher amount of sucralose in
circulation, and that measures should be taken to determine the
security of this occurrence Sylvetsky et al., 2017; Park et al., 2017.
These conflicting results just bring more doubt towards the con-
sumption of sucralose (see Fig. 15).
4.2. Natural intensive sweeteners
In recent years, the use of natural sweeteners has been
increasing, mainly due to demand from consumers. Even though
natural organisms like the EFSA or FDA do not differentiate
sweeteners by synthetic or natural, being all sweeteners regulated
by the EU normative 1129/2011.
The most common natural sweeteners are steviol glycosides
(E960), thaumatin (E957) and neohesperidine dihydrochalcone
(E959). Others, like tagatose and glycyrrhizin also exist in the
market but are not allowed to be used within the EU (Table 4).
Steviol glycosides (E960) are molecules extracted from the
leaves of Stevia rebaudiana Bertoni, a plant from the Asteraeae
family that is native from Paraguay and Portugal. The plant has such
a high concentration of steviosides that it can be directly used as a
sweetener (Lobete et al., 2017). The leaves are not allowed to be
used in the EU, but the purified, steviol glycosides compounds, are
(Periche et al., 2015). They are extracted with hot water and then
recrystallized in a hydroalcoholic solution. Steviol glycosides
consist of mixtures of different compounds, namely stevioside
(5e10%), rebaudioside A (2e5%), rebaudioside C (1%), dulcoside A
(0.5%), rebaudioside D, E and F (0.2%). Among them, the sweetest
compound is rebaudioside A (Gonzalez et al., 2014; Momtazi-
Borojeni et al., 2016). The combination of these molecules has a
sweetening power of over 300 times sucrose and an ADI limit of
4 mg kg
1 of body weight per day. These molecules are metabolized
by the colonic bacteria, converting them to steviol glucoronides to
finally be excreted through urine. In terms of caloric contribution, it
is neglectable, a thus safe for diabetic patients, while properties like
anti-inflammatory and diuretic effects are also attributed to the
compounds that comprise the mixture. Furthermore, it is relatively
stable to heat, can operate at a pH of 2e10. It displays a clean
sweetness, although some of the components have a bitter taste. A
A B
Fig. 15. Molecular structure of sucrose (A) the precursor of the sweetener sucralose (B).
311
small slice of the research concerning this sweetener has shown
concern about toxicity and genotoxicity of these compounds,
although many authors claim the database of in vitro and in vivo
studies is robust and there is no indication of the toxicity of ste-
vioside and rebaudioside (Barriocanal et al., 2008; Brusick, 2008;
Wheeler et al., 2008). Recently, concerns about the endocrine
disruption potential of steviol glucosides has become a hot topic,
with publications claiming there is a potential risk of these com-
pounds to have disrupting effects, suggesting further and deeper
research on this subject (Shannon et al., 2016; Urban et al., 2013,
2015). The uses of stevia encompass ice creams, yogurts, cakes,
sauces, drinks, bread, pastry, flavored milk, spices and as a tabletop
sweetener (Baines and Seal, 2012; Brandle et al., 1998; Carocho
et al., 2015; Nabors (Part I), 2001; Prakash and Chaturvedula,
2016). (See Fig. 16).
Thaumatin (E957) is also a mixture of compounds, namely
proteins, which are extracted from the Thaumatococcus danielli
Bennett plant, which is endemic to Africa (Baines and Seal, 2012). It
is a single-chain of 207 amino acid residues, which provide a sweet
taste at concentrations as low as 50 nM. There are other examples
of sweet proteins, like monellin, brazzein, and lysozyme, but
thaumatin is the most widespread example of protein-based
sweeteners (Beauchamp, 2016; Firsov et al., 2016). The extraction
is carried out with water and mechanical processes, being the most
important compounds Thaumatin A and B. In the plant, these
proteins display antimicrobial and protective functions, as sweet-
eners, they can be 2000 to 3000 times stronger than sucrose,
although the sweetness is quite slow to develop and has a residual
taste of licorice, thus being used in combination with other sugar
substitutes. In terms of stability, it has a high resistance to heat and
acidic pH, while being very soluble in water. Furthermore, it has no
caloric contribution and is very soluble in water and stable at high
temperatures (Baines and Seal, 2012; Pawar et al., 2013). It also has
a use as a flavor enhancer at a maximum of 0.5 mg kg
1, being used
in food commodities and supplements (EFSA, 2015). As a sweet-
ener, it is approved both in the EU (since 1984) and the US, where it
is considered GRAS. Soups, sauces, processed vegetables and egg-
derived products are the main foods where it is used. Given the
instabilities of its endemic region in West Africa, and climate
change, the production of thaumatin is not enough for demand,
therefore, many studies have focused on the production of re-
combinant thaumatin through microorganisms and transgenic
plants (Jain and Grover, 2015; Masuda, 2016; Nabors (Part I), 2001).
(See Fig. 17).
Neohesperidin dihydrochalcone (E959) is another intense semi-
natural sweetener that comes from the skin of the immature fruits
of Citrus aurantium L. When extracted, this compound is a flavone,
neohesperidin, which after suffering hydrolysis becomes a
312 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317

Table 4
Natural intense sweeteners, ADI, structure and sweetening power.

Name ADI (mg/kg/day) Brand Names Sweetening power

Thaumatin - E957 50 e 2000

Neohesperidine dihydrochalcone e E959 35 e 1500
Steviol glucosides - E960 4 Truvia® 300
PureVia®
Enliten®
Glycyrrhizin Not specified e 100
Tagatose Not considered a food additive e 0.92

C
A

Fig. 16. The most representative compounds of steviol glycosides, namely stevioside (A), rebaudioside A (B), rebaudiocide C (C), and dulcoside A (D).

dihydrochalcone (Baines and Seal, 2012). Other method to obtain
neohesperidin dihydrochalcone is through synthesis of naringenin,
extracted from the fruit of Citrus paradise Macfad. This sweetener is
reasonably hygroscopic and is stable at high temperatures, namely
during pasteurization, but quite insoluble, and presents an off-
white crystalline powder, although being poorly soluble in water
at room temperature but highly soluble in hot water (Nabors (Part
I), 2001). This fact is attenuated by the low quantities needed in
food, and the fact that it is used in combination with polyols or
glucose syrup. Its sweetening power, although quite high is far from
the most potent ones, achieving only 1500 times stronger than
glucose, and also has a very slow sweetening speed and a menthol
residue. One of its functions when used with other sweeteners is to
mask their unwanted tastes. It has a high ADI, of 35 mg kg
1 of body
weight per day and does not accumulate in tissues given to its quick
metabolization and excretion (EFSA, 2011). Approved in the EU
since 1994 but not in the US. This natural sweetener is also used to
thicken liquid foods, so it is used for ice creams, bubble gums,
pastry, water-based flavored drinks, milk and derivatives, snacks,
confectionary foodstuffs, beer, soups, food supplements and as a
tabletop sweetener and fruit derived food (El-Samragy, 2012;
Spillane, 2006). (See Fig. 18).
Glycyrrhizin, or more correctly glycyrrhizinic acid is a triterpe-
noid saponin that is obtained from the roots and rhizome of Gly-
cyrrhiza glabra, a plant known as licorice. It is described as being
from 30 to 200 times sweeter than sucrose. Apart from this, it is also
 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
Fig. 17. Representation of the protein thaumatin created with PYMOL software.
recognized for having several pharmacological and biological ac-
tivities, namely anticancer, anti-inflammatory, hepatoprotective,
antioxidant and antiviral. In the US, glycyrrhizin is considered GRAS
although there is a guideline for the maximum permitted levels for
the saponins on several preparations (Karkanis et al., 2016). In the
EU, although the Commission Report states that the consumption is
safe, a limit of 100 mg per day is recommended, given the gluco-
corticoid effects id the glycyrrhetinic acid present in the extract.
Still, given that the use of this extract is not widespread, these levels
are related to the consumption as licorice rather than a sweetener
(Komes et al., 2015). One drawbacks of using glycyrrhizin is the
potential hypertensive effects. Still, it has been used previously in
candies, chewing gum, toothpaste, beverages and tobacco.
Furthermore, its specific aroma and intense aftertaste are reasons
that it has not been very widespread (Graebin, 2016; Omar et al.,
2012). (See Fig. 19).
Tagatose is a ketohexose that is structurally similar to lactose,
displaying a hydroxyl group in C4, and can be found in small
quantities in fruits (cacao) and in dairy products. It is obtained from
lactose, though an enzymatic process followed by an isomerization
and purification steps. It displays a staggering 92% of sweetness
potency, but with only one third of the calories, making it inter-
esting for sweetening food (Bell, 2016; Jayamuthunagai et al., 2016).
Even though it is considered a sugar, it does not promote tooth
decay. Furthermore, tagatose, by having a different metabolization
to sucrose, does not interfere with blood glucose levels, deeming it
safe for diabetic individuals. Of all the consumed tagatose, only 20%
is absorbed by the intestine and readily excreted in urine (Ensor
Fig. 18. Chemical structure of neohesperidin dihydrochalcone.
313
Fig. 19. Representation of glycyrrhizin.
et al., 2014; Lu et al., 2007; Tandel, 2011; Levin, 2002). Although
tagatose is naturally occurring, its production is now industrialized
(through enzymatic reactions), making its classification as a semi-
synthetic compound in foodstuffs, and rendering a colorless crys-
talline powder with a bitter aftertaste, but compatible with a wide
range of food ingredients It has considerable advantage when
compared to other natural and synthetic sweeteners since it can be
considered a prebiotic and has a similar taste to sucrose (Dobbs and
Bell, 2010; Oh, 2007). It is stable at pH 2
7, displays a high solu-
bility in water, making it ideal to also be used as a flavor enhancer,
although it lacks some stability. Furthermore, it shows humectant
behavior like sorbitol, but decomposes faster than sucrose at high
temperatures. Tagatose is approved in New Zealand, Korea and in
the EU, although its use in the EU is not as a sweetener, rather as a
food ingredient. In the USA, it is considered GRAS and can be used
as a low-calorie sweetener. Its applications encompass beverages,
cereals, bubble gum, chocolate, caramels yogurts, ice creams,
nutritional supplements, and candy. Some conflicting results have
been found regarding its effects on gastrointestinal unrest. Geno-
toxic studies have been carried out and determined tagatose as
non-genotoxic (Baines and Seal, 2012; Kim, 2004; Nabors (Part I),
314 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
A to avoid these types of occurrences that just discredit both the in-
B avoid legislation gaps and harmonizing legislation across the many
Fig. 20. Lactose and tagatose.
2001). (See Fig. 20).
5. Conclusions and future perspectives
The WHO directives state that sugar should not represent more
than 10% of the daily caloric contribution, and is preparing to
propose a reduction to 5% in the near future (Mooradian et al.,
2017). This is a huge burden and at the same time an opportunity
for the food industry, governments and consumers! For one, the
opportunity to reduce the consumption of sugar is beneficial,
lowering the health issues associated with its excess consumption,
translating in a reduction of health spending by governments. On
the other hand, stronger and more effective sweeteners are
necessary to be added to food to carry out sweetening functions
without increasing the quantity of added sweetener, due to legal
restrictions and the alleged health implications of sweeteners.
The recent discovery that non-caloric sweeteners also seem to
increase the prevalence of diabetes and weight, unravels a grim
future for patients with sugar restrictions and the average con-
sumer, whom cannot be free from the dangers of eating? Or is
research “simply just seeking to deep?” Is research trying to find, at
all costs, culprits for sicknesses, and links between sweeteners and
illnesses (Fowler et al., 2008; Imamura et al., 2015; de Koning et al.,
2011)? Could some of these metabolic disorders happen to be, like
cancer, which recent studies state that two thirds of carcinomas are
a matter of “luck” rather than lifestyle or gene disorders? What will
be the fate of eating in the beginning of the century? (Nowak and
Waclaw, 2017; Tomasetti and Vogelstein, 2015).
Further entropy arises when similar studies end up in conflict-
ing results. One incredible example is detailed by Bes-Rastrollo
et al. (2013), in which similar studies found contradicting results.
In one, funded by food industry companies, in 83% of studies stated
that insufficient proof was detected for a correlation among the
consumption of sugar-sweetened beverages and weight gain, while
in other studies, without industrial financing, the same percentage
could relate the increase in weight with the consumption of such
beverages. Even before this study, uncomfortable positions have
been identified by researcher, especially clinical investigators when
carrying out their research while being funded by enterprises. The
risk of biased reports in the food industry is continuous, given the
different ways associations of enterprises and researchers can be
achieved, namely through sponsorships of journals and confer-
ences, sponsorships of research studies, partnerships and alliances.
Although there is always a latent risk in virtually all areas, because
advances in industry come from research, tight regulation is critical
dustry and research (Boyd et al., 2003; Nestle, 2001).
After a “not so peaceful” coexistence between consumers and
sweeteners, fueled by scandals, legislation gaps and controversies,
and occasional deception by the industry, the public wants to be
assured that they are consuming safe sweeteners. There are many
forms to induce trust in consumers, but they rely on combined
efforts from the industry, governments, press/media and con-
sumers. The industry must, at all costs research and produce safe,
sound, cheap, easily producible, sustainable and strong sweeteners.
Governments must act in two manners. Firstly, combining efforts to
governing bodies (EFSA, FDA …), which can be an incredible leap in
gaining consumer trust. In the XXI century, it is unthinkable that
cyclamates and neohesperidine dihydrochalcone are allowed in the
EU and not in the US, while D-tagatose is not considered a sweet-
ener in the EU and cannot be used as one, if the scientific back-
ground is available for both governing bodies to consult and
legislate accordingly. Why the legislation difference? The second
effort that could be put forward by governments is the public ed-
ucation regarding eating, healthy lifestyles and choices. The press
and media play a role in the scandals, speculation and exaggeration
of studies and legislation, and should tone down when dissemi-
nating information that can be badly interpreted by the public,
especially the uninformed fringe. Finally, the consumer must, at all
costs seek trustworthy information to make informed decisions
about what they are eating. In the era where information is
everywhere, finding reliable information can be the difficulty, but
scientific publications are somewhat easier to find as they tend to
become open source. Reading the article or study that originated a
news article is always better than to read the news itself, for the
scientific reports are stripped from exaggeration and extrapolation,
conveying specific findings.
What are the next generation of sweeteners, and how are they
going to affect the population is the question to be asked. There
should be a constant seek for new low caloric sweeteners, namely
isomers of known ones and other related compounds that have
should have higher sweetening powers, lower production costs,
lower caloric contribution and no health impact. Concomitantly,
the major investment should be on both synthetic and natural non-
caloric sweetener. The synthetic molecules will be rearrangements
of known sweeteners or prospection for new ones, relying on
combinatorial chemistry, but also the improvement of the pro-
duction of widespread sweeteners. The answer to passing the idea
of safety could be the production of these compounds through
ecological and sustainable processes, translating in a better un-
derstanding of the public, while associating sweeteners with eco-
friendly and safety ideas. The marketing of studies that deem
artificial sweeteners safe is critical to, through scientific discovery
inform consumers that they are consuming safe chemicals. The
natural sweeteners financing will be for better extraction methods
of currently impracticable compounds and the discovery of new
sources of sweetening molecules. Natural sweeteners have been
becoming more widespread due to the public's perception that
what is natural is less hazardous for health, and both manufactures
and food companies have started to shift considerable amounts of
funds to new natural sweeteners.
Further innovations in the sector could arise from new types of
sweeteners or improvements. Compounds like miraculin, which is
not a sweet molecule, but alters taste buds to perceive food as
sweet could be improved while, other similar molecules can be
found in nature or synthesized and used in the food industry. These
molecules or extracts could be added to food to make it be
perceived as sweet, rather than making them actually sweet, ulti-
mately avoiding alleged health impacts from sweeteners.
 M. Carocho et al. / Food and Chemical Toxicology 107 (2017) 302e317
Although some improvements and innovations may seem
temporally closer than others, the truth is that the sweeteners
market, mainly regarding sweeteners as food additives, will be
suffering changes and having to tackle health and economic issues,
while coping with regulation that should become more harmonic
to ensure trust and peace of mind for consumers, because the
reduction of sugar intake seems imminent and necessary. By 2020,
EFSA will be providing scientific advice of added sugar in food,
establishing a science-based cut-off value for daily exposure to
added sugars from all sources which is not associated with adverse
health effects. This means that the concerns about sugar's effects on
health are real, making this an opportunity for the sugar substitutes
to become safer, trustworthy, and, expectedly sweeter.
Conflict of interest
The authors state no conflicts of interest regarding the
manuscript.
Acknowledgments
The authors are grateful to the Foundation for Science and
Technology (FCT, Portugal) and FEDER under Programme PT2020
for financial support to CIMO (UID/AGR/00690/2013). Author
Ma
rcio Carocho also thanks FCT for his post-doctoral grant SFRH/
BPD/114650/2016.
Transparency document
Transparency document related to this article can be found
online at http://dx.doi.org/10.1016/j.fct.2017.06.046.
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