Phytochemistry 64 (2003) 913–921

www.elsevier.com/locate/phytochem

Molecules of Interest

Stevioside
Jan M.C. Geuns*
Laboratory of Plant Physiology, Catholic University of Leuven, Kasteelpark Arenberg 31, B 3001 Leuven, Belgium

Accepted 18 June 2003

Abstract
Stevioside is a natural sweetener extracted from leaves of Stevia rebaudiana (Bertoni) Bertoni. The literature about Stevia, the
occurrence of its sweeteners, their biosynthetic pathway and toxicological aspects are discussed. Injection experiments or perfusion
experiments of organs are considered as not relevant for the use of Stevia or stevioside as food, and therefore these studies are not
included in this review. The metabolism of stevioside is discussed in relation with the possible formation of steviol. Different
mutagenicity studies as well as studies on carcinogenicity are discussed. Acute and subacute toxicity studies revealed a very low
toxicity of Stevia and stevioside. Fertility and teratogenicity studies are discussed as well as the effects on the bio-availability of
other nutrients in the diet. The conclusion is that Stevia and stevioside are safe when used as a sweetener. It is suited for both
diabetics, and PKU patients, as well as for obese persons intending to lose weight by avoiding sugar supplements in the diet. No
allergic reactions to it seem to exist.
# 2003 Elsevier Ltd. All rights reserved.
Keywords: Stevia rebaudiana; Stevioside; Sweetener; Biosynthesis; Toxicology

1. Introduction depending on the cultivar and growing conditions. Ste-

vioside 3 is the main sweet component. Other com-
Stevia rebaudiana (Bertoni) Bertoni is a perennial pounds present but in lower concentration are:
shrub of the Asteraceae (Compositae) family native to steviolbioside 2, rebaudioside A 4, B 5, C 6, D 7, E 8, F
certain regions of South America (Paraguay and Brazil). 9 and dulcoside A 10 (Kennelly, 2002; Starrat et al.,
It is often referred to as ‘‘the sweet herb of Paraguay’’. 2002). The presence of steviolbioside and rebaudioside
Stevioside, the main sweet component in the leaves of B in extracts might be due to artifacts of the extraction
Stevia rebaudiana (Bertoni) Bertoni tastes about 300 procedure (Refs. in Kennelly, 2002).
times sweeter than sucrose (0.4% solution). Structures Details on the genus Stevia, its botany, its sweet and
of the sweet components of Stevia occurring mainly in non-sweet constituents, modifications of the naturally
the leaves are given in Fig. 1. Their content varies occurring sweeteners to improve the taste can be found
between 4 and 20% of the dry weight of the leaves in the recent excellent book by Kinghorn (2002). Both
the Stevia plant, its extracts, and stevioside have been
used for several years as a sweetener in South America,
Abbreviations: ADI, Allowable daily intake; BW, Body weight; Asia, Japan, China, and in different countries of the
CHL, Chinese hamster lung fibroblast cell line; GA, gibberellin; Glc, EU. In Brazil, Korea and Japan Stevia leaves, stevioside
Glucose; ICH, International Council of Harmonisation; DXR, 1- and highly refined extracts are officially used as a low-
deoxy-d-xylulose-5-phosphate reductoisomerase; DXS, 1-deoxy-d-
xylulose-5-phosphate synthase; EST, expressed sequence tags; IPP, calorie sweetener (Mizutani and Tanaka, 2002; Kim et
isopentenyldiphosphate; JECFA, Joint FAO/WHO Expert Committee al., 2002). In the USA, powdered Stevia leaves and
on Food Additives; LD50, Lethal dose at which 50% of the animals refined extracts from the leaves have been used as a
die; MEP, 2-C-Methyl-d-erythritol-4-phosphate; MVA, mevalonic dietary supplement since 1995. In 2000, the European
acid; NOEL, No-observable effect level; OECD, Organisation for Commission refused to accept Stevia or stevioside as a
economic co-operation and development; PKU, Phenylketonuria;
Rha, Rhamnose; Xyl, Xylulose
novel food because of a lack of critical scientific reports
* Tel.: +32-16-321510; fax: +32-16-321509. on Stevia and the discrepancies between cited studies
E-mail address: jan.geuns@bio.kuleuven.ac.be (J.M.C. Geuns). with respect to possible toxicological effects of stevioside
0031-9422/03/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0031-9422(03)00426-6
914 J.M.C. Geuns / Phytochemistry 64 (2003) 913–921

Fig. 1. Structures of stevioside and related compounds. In rebaudioside D and E R1 is composed of 2 b-Glc-b-Glc(2!1). In rebaudioside A, B, C,
D, E and F in group R2 an additional sugar moiety is added on carbon 3 of the first b-Glc. In rebaudioside F one b-Glc is substituted for by-b-Xyl.

and especially its aglycone steviol 1 (Fig. 1) (Kinghorn, plant DXS and DXR enzymes. Furthermore, it was
2002; Geuns, unpublished). The advantages of stevio- demonstrated through heterologous expression in
side as a dietary supplement for human subjects are Escherichia coli that the cloned cDNAs encode func-
manifold: it is stable, it is non-calorific, it maintains tional proteins (Totté et al., 2003). Kim et al. (1996a)
good dental health by reducing the intake of sugar and found a high activity of HMG-CoA reductase, a key
opens the possibility for use by diabetic and phenylke- enzyme of the mevalonic acid (MVA) route to IPP, in
tonuria patients and obese persons. the chloroplast fraction from Stevia. They suggested
that steviol was synthesised via MVA, but no direct
proof was given to support this assessment. The results
2. Biosynthesis of stevioside concerning the involvement of the MEP pathway in the
biosynthesis of steviol cast doubt on their hypothesis as
The ent-kaurene skeleton of stevioside and hence also no contribution of the MVA pathway could be detected.
of gibberellins (GAs) is formed via the recently dis- These conclusions were confirmed by Brandle et al.
covered 2-C-Methyl-d-erythritol-4-phosphate pathway (2002) who sequenced 5548 expressed sequence tags
(MEP; Totté et al., 2000). The genes of the enzymes (ESTs) from a Stevia leaf cDNA library. The ESTs were
catalysing the first two steps of this pathway, 1-deoxy-d- classified according to their function in primary or sec-
xylulose-5-phosphate synthase (DXS) and 1-deoxy-d- ondary metabolism. In the last category many candidate
xylulose-5-phosphate reductoisomerase (DXR) were genes specific to the MEP pathway and no members of
cloned using reverse transcriptase-PCR. DXS and DXR the MVA pathway were identified, suggesting that the
from Stevia both contain an N-terminal plastid target- primary source of IPP for diterpene biosynthesis is via
ing sequence and show high homology to other known the MEP pathway. A kaurene oxidase, a cyt P450 type
 J.M.C. Geuns / Phytochemistry 64 (2003) 913–921 915

of enzyme, was highly represented. Other P450s respectively male and female animals. In rats and mice
involved in GA biosynthesis, like ent-kaurenoic acid the LD50 was above 15 g/kg BW demonstrating that of
oxidase, were not found among the ESTs. These results the tested animals hamsters are much more sensitive to
demonstrate the divergence of steviol and GA steviol (Toskulkao et al., 1997).
biosynthesis after the production of kaurenoic acid. In chronic subacute toxicity studies with rats during 3
Kim et al. (1996b) claimed to have purified and partially months (Mitsuhashi, 1976; Akashi and Yokoyama,
characterized the ent-kaurenate 13-hydroxylase, the key 1975; Aze et al., 1991) or with hamsters over several
enzyme in steviol biosynthesis. However, use of the pub- generations (Yodyingyuad and Bunyawong, 1991) a
lished N-terminal sequence for constructing primers led NOEL higher than 2.5 g/kg BW was found. From this an
to fructose-bisphosphate aldolase instead of an ent-kaur- ADI of 25 mg/kg BW can be deduced (safety factor 100).
enate 13-hydroxylase (Totté, personal communication).
Looking at the accumulation percentage of the sweet
glycosides, it is clear that a large fraction of total plant 4. Steviol, the controversial metabolite of stevioside
metabolism is committed to the synthesis of these
structurally complex molecules. In contrast, gibberellins Mutagenic effects of steviol, the aglycone of stevio-
such as GA20 are present in Stevia leaves at concentra- side, and/or its metabolites were reported in Salmonella
tions of 1.2 mg/kg fresh weight, over 10,000 times lower typhimurium TM677 (Pezzuto et al., 1985; Compadre et
than steviol glycosides (Alves and Ruddat, 1979). How al., 1988; Matsui et al., 1996a; Temcharoen et al., 1998;
can these differences in the occurrence of such structu- Terai et al., 2002). After metabolic activation it was
rally related products be explained? Richman et al. shown that so far unknown steviol metabolites caused
(1999) concluded that profound changes in the regula- mutations in Salmonella typhimurium TM677, i.e. tran-
tion of copalylphosphate synthase and kaurene synthase sitions, transversions, duplications and deletions at the
expression in Stevia leaves have enabled the synthesis guanine phosphoribosyltransferase (gpt) gene (Matsui,
and accumulation of high concentrations of sweeteners. 1996b). However, stevioside and steviol were inactive in
The fact that expression levels are highest in mature various TA strains of Salmonella typhimurium, Escher-
tissues as compared to young rapidly growing tissues ichia coli WP2 uvrA/pKM101 and the rec-assay using
raises the possibility of temporal and spatial separation, Bacillus subtilis even when activated microsomal frac-
preventing an overlap of steviol and GA biosynthesis. tion was present (Matsui et al., 1996a; Klongpanichpak
Various plant-growth regulator effects, mostly gib- et al., 1997). The direct mutagenic activity of 15-oxo-
berellin-like, have been described of stevioside, steviol steviol was refuted by Procinska et al. (1991), but con-
and isosteviol or of their metabolites (Hersenhorn et al., firmed by Terai et al. (2002). The activity of steviol in
1997; de Oliveira et al., 1999; Bearder et al., 1975; Salmonella typhimurium TM677 was very low and was
Gianfagna et al., 1983; Orihara et al., 1991; Ruddat et only about 1/3000 that of 3,4-benzopyrene and that of
al., 1963; de Oliveira and Strapasson, 1996). steviol methyl ester 8,13 lactone was 1/24,500 that of
furylfuramide (Terai et al., 2002). Although a weak
activity of steviol and some of its derivatives was found
3. Acute and chronic toxicity in the very sensitive S. typhimurium TM677 strain, the
authors concluded that the daily use of stevioside as a
The toxicology and safety of stevioside used as a sweetener is safe. Moreover, the presence in the blood of
sweetener were recently reviewed (Geuns, 2002; Hux- the chemically synthesised steviol derivatives after feed-
table, 2002). An acceptable daily intake (ADI) of 7.9 mg ing stevioside is not proven at all. Very high doses of
stevioside/kg BW was calculated (Xili et al., 1992). steviol (90% purity) intubated to hamsters (4 g/kg bw),
However, this ADI should be considered as a minimum rats and mice (8 g/kg BW) did not induce micronucleus
value as the authors did not test concentrations of ste- in bone marrow erythrocytes of both male and female
vioside higher than 793 mg/kg BW. animals. However, these doses showed some cytotoxic
Neither those scientific studies where Stevia extract or effect to the female, but not to the male of all treated
solution of pure stevioside were injected in animals, nor animal species (Temcharoen et al., 2000). It is not
those studies employing perfusion experiments of excluded that the toxicity is due to the 10% impurities
organs, are considered relevant for the use of Stevia or present. After metabolic activation of steviol some gene
stevioside as a food additive and are not discussed in mutation and chromosomal aberration was found in
this review. Stevioside has a very low acute oral toxicity Chinese hamster lung fibroblasts (Matsui et al., 1996a).
in the mouse, rat and hamster. An oral LD50 between It has to be said that of all animals tested hamsters had
8.2 and 17 g/kg BW was found (Mitsuhashi, 1976; the most sensitive response. Moreover, in hamster sev-
Medon et al., 1982; Toskulkao et al., 1997). eral metabolites of stevioside were found that are not
In hamsters the LD50 of steviol (90% purity), the formed in rats or humans. Therefore, the relevance of
aglycone of stevioside, was 5.2 and 6.1 g/kg BW for experiments with hamsters should be questioned.
916 J.M.C. Geuns / Phytochemistry 64 (2003) 913–921

5. Uptake and metabolism of stevioside 15a-hydroxysteviol and steviolbioside. It is not possible

to deduce the amounts of metabolites from the chro-
It has been shown that oral stevioside is not taken up matograms given. As all the extract fractions were trea-
by the human body or the uptake is extremely low ted with type H-5 b-glucuronidase/sulfatase at 55 ! C for
(Yamamoto et al., 1985; Bracht et al., 1985; Koyama et 3 h some of the metabolites might possibly originate
al., 2003b; Geuns et al., in press a) and none of the from this treatment. For steviol was the only metabolite
digestive enzymes from the gastro-intestinal tract of of stevioside when bacteria from the hamster caecum
different animals and man are able to degrade stevioside were incubated (Hutapea et al., 1997). Moreover, it is
into steviol, the aglycone of stevioside (Wingard et al., unlikely that compounds as steviol-16,17a-epoxide or
1980; Hutapea et al., 1997; Koyama et al., 2001, 2003a). 15a-hydroxy-steviol are formed in the anoxic intestines
Nevertheless, in feeding experiments with rats and of hamsters. Isosteviol might be an artifact due to acid
hamsters stevioside was metabolised to steviol by the conditions and is normally not detected even not after
bacterial flora of the caecum. Steviol was found in the incubating stevioside in human stomatal juice for 6 h
blood of the animals with the maximum concentration (Geuns, unpublished). The peaks in the chromatograms
occurring after 8 h (Nakayama et al., 1986; Koyama et of plasma and urine identified as stevioside were impu-
al., 2003a). In the cited studies, it was not indicated that rities cochromatographing at the same RT as shown
coprophagy, occurring in rodents, was prevented, so it later (Hutapea, personal communication).
is not clear whether the steviol occurring in the blood When steviol was fed to rats is was easily taken up by
was taken up directly from the colon or indirectly from the intestines (Wingard et al., 1980; Nakayama et al.,
the ingested faeces (after passing through the intestines 1986; Koyama et al., 2003b). The easy uptake of steviol
again). Although bacteria isolated from the human by the gastro-intestinal tract was demonstrated in
colon are able to transform stevioside into steviol in experiments with everted gastrointestinal sacs (Koyama
vitro (Hutapea et al., 1997; Koyama et al., 2001, 2003a; et al., 2003b) and Caco-2 cell monolayers (Geuns et al.,
Gardana et al., in press), it has never been proven that in press a). Although it was demonstrated that the
this is also the case in vivo nor that the steviol possibly absorptive transport of steviol was high in Caco-2 cell
formed in the colon is taken up directly from it. More- monolayers, and that stevioside fed to pigs (68 mg/kg
over, studies with roosters (Pomaret and Lavieille, 1931) BW) was completely converted in the colon into steviol,
and chickens (laying hens and broylers, Geuns et al., no steviol could be detected in the blood of the animals,
2003) indicated that stevioside was rapidly eliminated suggesting that the possible uptake from the colon is
from the body, largely untransformed. Opposed to these very low (Geuns et al., in press a). The lack of steviol in
results, in pigs oral stevioside was completely degraded the blood samples can probably not be attributed to
into steviol that was the only metabolite in the faeces. metabolism during or after uptake as was the case with
However, no stevioside or steviol were found in the soy isoflavones that after uptake were metabolised to
blood (Geuns et al., in press a). From the references compounds that were hydrolyzable with a combined b-
cited above it is concluded that only the bacteria from glucuronidase and sulfatase enzyme preparation (Setch-
the caecum or colon were able to degrade stevioside into ell, 2002); indeed free steviol was detected in the plasma
steviol (caecum of mice, rats, hamsters and chickens; of rats up to at least 8 h after feeding stevioside or ste-
colon of pigs and man). In one experiment, the bacteria viol (Compadre et al., 1988; Koyama et al., 2003b).
from the human colon also formed steviol epoxid in Moreover, the in vitro conversion of steviol by liver
vitro, that was again metabolised to steviol (Hutapea et microsomal fraction from Aroclor 1254-pretreated rats
al., 1997). However, in vivo this epoxid formation most was rather low (about 0.3% after 2 h, Compadre et al.,
probably will not occur due to the anaerobic conditions 1988). The intrinsic clearance of steviol by human
of the human colon. It was correctly concluded that microsomes was about 4 times lower than that of rat
steviol is the only metabolite in faeces and is not further microsomes (Koyama et al., 2003b).
metabolised (Hutapea et al., 1997; Koyama et al., 2001, Taking into account the very low detection limits of
2003a; Gardana et al., in press; Geuns et al., 2003, in steviol (50 pg) when analysed as the (7-methoxy-
press a). Anyway, steviol epoxid has been tested in coumarin-4-yl)methyl ester the amount of steviol possi-
mutagenicity studies and showed to be inactive (Pezzuto bly remaining undetected in blood samples of pigs fed
et al., 1985). In contrast to the above studies is the work stevioside can be estimated to be very low (below 1 mM,
by Hutapea et al. (1999) who found, besides stevioside i.e. below 318 ng/ml; Geuns et al., in press a). This
and steviol, steviol-16,17a-epoxide, 15a-hydroxysteviol, hypothetical maximum steviol concentration in the
isosteviol and steviolbioside in the faeces 24 h after blood would probably not be toxic, as in hamsters fed
force-feeding hamsters with 1 g stevioside/kg BW. In the steviol up to 250 mg/kg BW no toxic effects were found
urine steviol-16,17a-epoxide, stevioside, 15a-hydro- (Wasuntarawat et al., 1998). In this case steviol would
xysteviol, steviolbioside and isosteviol were found. The have been easily taken up by the intestines. When steviol
plasma contained steviol-16,17a-epoxide, stevioside, was fed to rats (45 mg/kg BW) a fast uptake was found
 J.M.C. Geuns / Phytochemistry 64 (2003) 913–921 917

Table 1
Steviol concentration (mg/ml plasma) measured in the blood of rats (Koyama et al., 2003b) or estimated to be present in the blood of hamsters
(Wasuntarawat et al., 1998) and pigs (Geuns et al., in press a) after the administration of steviol to the animalsa

Amount administered After

(mg/kg BW)
15 min (mg/ml) 2 h (mg/ml) 4 h (mg/ml) 8 h (mg/ml) 16 ha (mg/ml)

45 (rats) 18.3 2.5 3.5 2.5 1

250 (hamsters) 102 13.6 19 13.6 5.4
9.7 (pigs) 3.94 0.5 0.7 0.5 0.2
a
Amount estimated from the decay of the proceeding data points.

and the highest plasma concentration of 18.3 mg/ml was refuted by Shiotsu (1996) who did more reliable experi-
observed after 15 min (the first data point, Table 1; ments with many more animals using methods as simi-
Koyama et al., 2003b). The plasma concentration lar as possible to the methods used by Planas and Kuc.
declined to about 2–3 mg/ml at 8 h. Although we are No effect on general condition, body weight, water con-
aware that species differences might occur, we have sumption, live birth rate or litter size was found. No
extrapolated the data obtained in rats to hamsters and effects of stevioside were found on fertility or reproduc-
pigs (Table 1). Assuming a similar uptake and metabo- tion in mice (Akashi and Yokoyama, 1975), rats (Mori et
lism in hamsters the reported NOEL of 250 mg steviol/ al., 1981; Xili et al., 1992; Sinchomi and Marcorities,
kg BW would correspond to a plasma concentration of 1989) or hamsters (Yodyingyuad and Bunyawong, 1991).
102 mg/ml 15 min after intubating steviol and about 13.6 No significant effect was found on spermatogenesis,
mg/ml after 8 h. In pigs having at the most 9.7 mg ste- nor on the interstitial cell proliferation and tumor for-
viol/kg BW available in the colon for uptake, this would mation in the testes of F344 rats fed a ration containing
then be 3.94 and 0.5 mg/ml respectively (Geuns et al., in up to 1% stevioside (95.2% purity) for 22 months
press a). Although these concentrations are above the (Yamada et al., 1985).
detection limit of the (7-methoxycoumarin-4-yl)methyl Whereas Melis (1999) suggested a possible decrease of
ester of steviol, no steviol was detected in the blood of the fertility of male rats by a very high dose of Stevia
these animals. Therefore, it was suggested that in vivo extract, Oliveira-Filho et al. (1989) who administered
the uptake of the carboxy acid steviol from the colon is extracts with similar stevioside content stated that there
neglectible and that it is rather adsorbed to the com- is certainly not an effect on male fertility. It is not sure
pounds present in the colon (pH 7–7.5) of which the that the observed effects were due to the stevioside pre-
contents is being concentrated by withdrawal of water. sent in the extract. It should also be mentioned that the
used extract concentrations were extremely high, at the
start of the experiments even 5.34% of the body weight
6. Stevioside and carcinogenicity (or around 5.3 g stevioside/kg bw). For an adult person
of 65 kg this means 3.47 kg of dry Stevia leaves or about
A weak mutagenic effect of steviol (only 90% purity) in 34.7 kg fresh leaves/day, i.e. more than 50% of the body
one sensitive Salmonella typhimurium TM 677 strain (see weight! The significance of such experiments where only
above) does not mean that stevioside used as a sweetener one extremely high concentration was tested, should be
should be carcinogenic in se, even if the stevioside might questioned. Melis’ results are also in contradiction with
be transformed to steviol by bacteria in the colon. The the above and below cited studies that could not reveal
safety of oral stevioside in relation to carcinogenic activ- any effect on fertility of male or female animals.
ity is evidenced by the work of Yamada et al. (1985), Xili Chicken embryos react very sensitively to adminis-
et al. (1992), Toyoda et al. (1997) and Hagiwara et al. tered toxicants. Fertile broiler eggs (Ross) were injected
(1984) with rats. Very significant inhibitory effects of ste- with stevioside or steviol (Geuns et al., in press b). At
vioside were reported on tumor promotion by 12-O- hatch (day 21) and 1 week later no influence of the dif-
tetradecanoylphorbol-13-acetate in carcinogenesis in ferent treatments could be found on embryonic mortal-
mouse skin (Yasukawa et al., 2002). In 1999 the JECFA ity, body weight of the hatchlings, deformations (e.g.
clearly stated that there was no indication of carcinogenic bone, beak and head malformations, abnormal feather-
potential of stevioside (WHO, 1999). ing, open vent) or abnormal development of the gonads.
The hatchlings developed normally. It was concluded
that prenatal exposure to stevioside and steviol was not
7. Fertility and teratogenicity toxic to the chicken embryo.
Applied stevioside has no effect on fertility, mating
The results of a decrease of live birth rate in rats performance, pregnancy, number of fetuses, nor on the
(Planas and Kuæ, 1968) by Stevia decoctions were growth and fertility of the offspring (Yodyingyuad and
918 J.M.C. Geuns / Phytochemistry 64 (2003) 913–921

Bunyawong, 1991; Mori et al., 1981; Oliveira-Filho et Melis, 1996; Xili et al., 1992; Oliveira-Filho et al., 1989;
al., 1989; Sinchomi and Marcorities, 1989; Usami et al., Das et al., 1992) and hamsters (Yodyingyuad and
1995; Geuns et al., in press b). However, when steviol Bunyawong, 1991).
(the aglucone of stevioside) was given to hamsters on In all of the above cited experiments, no indications
days 6–10 of pregnancy at doses of 500–1000 mg/kg of any influence on the bio-availability of nutrients, nor
body weight/day it induced toxicity (Wasuntarawat et on physiological effects were found. The animal feed
al., 1998). The number of live foetuses per litter and used in the experiments did not contain sugar supple-
mean foetal weight decreased. The maternal kidneys ments. Therefore, no reduction of weight gain was
showed a dose-dependent increase in severity of con- observed in the experiments as Stevia or stevioside did
voluted tubules in the kidneys. This study with steviol not substitute for added sugar.
has nothing to do with the use of stevioside as a
sweetener. When stevioside is fed to hamsters, no toxic
effects were found even not in 3 successive generations 9. Stevia, stevioside and special groups of the
(Yodyingyuad and Bunyawong, 1991). When steviol is population: nutritional significance
given in the feed, it can be resorbed directly by the
intestines, whereas stevioside is not. Moreover, ham- Boeckh-Haebisch (1992) concluded that concentrated
sters are known to be very sensitive to steviol and Stevia extracts in normal doses to sweeten can be used
stevioside (Toskulkao et al., 1997), this is the reason without restriction by normal persons as well as by dia-
that hamsters were chosen in this study. The NOEL betics. The omission of excessively added sugar in the
of steviol was 250 mg/kg bw (Wasuntarawat et al., food is beneficial to diabetics by lowering the blood
1998), which corresponds to 625 mg stevioside/kg bw. sugar content (Boeckh-Haebisch, 1992). Stevia and ste-
Even under these very unfavourable conditions an vioside are also safe for phenylketonuria (PKU)
ADI of 2.5 mg steviol/kg bw, which corresponds to patients as no aromatic amino acids are involved. Obese
6.25 mg stevioside/kg bw, can be calculated, which is persons might lose weight by the fact that excessive
close to 7.9 mg/kg bw obtained for stevioside (Xili et sugar in the food is replaced by Stevia or stevioside.
al., 1992). Omitting added sucrose in foods increases the relative
proportion of polymeric carbohydrates. This has a ben-
eficial effect for a balanced food intake and for human
8. Bio-availability of nutrients from the diet health (Anonymous, 1996).
Curi et al. (1986) reported that Stevia extracts from 5
Modern broiler chickens are intensively selected for g of dried leaves thrice a day administered for 3 days to
growth rate and BW increases with a factor of more healthy volunteers lowered the plasma glucose levels.
than 50 in a time span of 6 weeks, making these ani- However, care should be taken interpreting these results
mals especially suited to study the influence of feed as the plasma glucose level of the Stevia treated group
additives on growth. However, they have become very was already significantly lower before the administra-
susceptible to even slight deviations from optimal tion of the extract. Intravenous administration of ste-
environmental and nutritional conditions. If such vioside (95% pure, 50, 100 or 200 mg/kg BW) resulted
aberrations occur, this is readily reflected in feed intake in a significant hypotensive effect in spontaneously
and growth rate. No effects of stevioside on the hypertensive rats without adverse effects on heart rate
growth, feed uptake or feed conversion of broiler or serum catecholamine levels (Chan et al., 1998). In a
chickens were found (Wood et al., 1996; Geuns et al., study with humans stevioside (250 mg thrice a day) was
2003). It can be inferred that stevioside did not influ- administered for 1 year to 60 hypertensive volunteers
ence the uptake of other essential nutrients such as (Chan et al., 2000). After 3 months the systolic and
amino acids, vitamins, minerals etc. When stevioside diastolic blood pressure significantly decreased and the
was supplemented to the feed of laying hens (667 mg/ effect persisted during the whole year. Blood biochem-
kg), no significant differences were found for the total istry parameters including lipid and glucose showed no
feed consumption, body weight gain, the total egg pro- significant changes. No significant adverse effect was
duction nor for the feed conversion calculated as the observed and quality of life assessment showed no
ratio between total feed uptake and total gram of egg detoriation. The authors concluded that stevioside is a
mass produced during the 10 days of the experiment well tolerated and effective compound that may be
(Geuns et al., 2003). The percentage of yolk and egg considered as an alternative or supplementary therapy
white was not significantly different between the control for patients with hypertension. Liu et al. (2003) repor-
and the stevioside-treated group. These results showing ted that the underlying mechanism of the hypotensive
the lack of effects on growth and hence on bioavail- effect of administered stevioside in dogs (200 mg/kg
ability of essential nutrients are in good accordance BW) was due to inhibition of Ca2+ influx from extra-
with studies performed with rats (Yamada et al., 1985; cellular fluid.
 J.M.C. Geuns / Phytochemistry 64 (2003) 913–921 919

10. Stevioside and caries Das, S., Das, A.K., Murphy, R.A., Punwani, I.C., Nasution, M.P.,
Kinghorn, A.D., 1992. Evaluation of the Cariogenic Potential of the
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Acknowledgements Metabolism of stevioside in pigs and intestinal absorption char-
acteristics of Stevioside, Rebaudioside A and Steviol. Food Chem.
The author acknowledges the ‘‘Onderzoeksraad Toxicol. (in press a).
KULeuven’’ for grant OT/00/15, the FWO for grant Geuns, J.M.C., Malheiros, R.D., Moraes, V.M.B., Decuypere, E.M.-
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