1  

Sweetness and sensory properties of commercial and novel oligosaccharides of 1 

prebiotic potential 2 

3 

Laura Ruiz-Aceitunoa, Oswaldo Hernandez-Hernandeza, Sofia Kolidab, F. Javier 4 

Morenoa,* and Lisa Methvenc 5 

6 

a Institute of Food Science Research, CIAL (CSIC-UAM), Nicolás Cabrera 9, 28049 7 

Madrid (Spain) 8 

b OptiBiotix Health plc, Innovation Centre, Innovation Way, Heslington, York YO10 9 

5DG (UK) 10 

c Sensory Science Centre, Department of Food and Nutritional Sciences, The University 11 

of Reading, PO Box 226, Whiteknights, Reading RG6 6AP (UK) 12 

13 

*Corresponding author: javier.moreno@csic.es Tel (+34) 91 0017948 14 

2  

Abstract 15 

This study investigates the sweetness properties and other sensory attributes of ten 16 

commercial and four novel prebiotics (4-galactosyl-kojibiose, lactulosucrose, lactosyl-17 

oligofructosides and raffinosyl-oligofructosides) of high degree of purity and assesses the 18 

influence of their chemical structure features on sweetness. The impact of the type of 19 

glycosidic linkage by testing four sucrose isomers, as well as the monomer composition 20 

and degree of polymerization on sweetness properties were determined. Data from the 21 

sensory panel combined with principal component analysis (PCA) concludes that chain 22 

length was the most relevant factor in determining the sweetness potential of a 23 

carbohydrate. Thus, disaccharides had higher sweetness values than trisaccharides which, 24 

in turn, exhibited superior sweetness than mixtures of oligosaccharides having DP above 25 

3. Furthermore, a weak non-significant trend indicated that the presence of a ketose sugar 26 

moiety led to higher sweetness. The novel prebiotics tested in this study had between 18 27 

and 25% of relative sweetness, in line with other commercial prebiotics, and samples 28 

varied in their extent of off flavour. Therefore, these findings suggest a potential use for 29 

clean tasting prebiotics as partial sugar replacers, or in combination with high intensity 30 

sweeteners, to provide a well-balanced sweetness profile. 31 

32 

Keywords: sweetener; enzymatic synthesis; sensory evaluation; free sugar substitute; 33 

non-digestible oligosaccharides. 34 

3  

1. Introduction 35 

A high level of free sugars intake is associated with poor dietary quality, dental caries, 36 

obesity or diabetes among other noncommunicable diseases (WHO/FAO Expert 37 

Consultation, 2003). Free sugars are defined as monosaccharides and disaccharides added 38 

to foods and beverages and sugars naturally present in honey, syrups, fruit juices and fruit 39 

juice concentrates. In 2015, the World Health Organization (2015) published a guideline 40 

on sugar intake for adults and children where the main and strong recommendation was 41 

to reduce the intake of free sugars to less than 10% of total energy intake, with a 42 

conditional recommendation for further reduction to below 5% of total energy intake. 43 

Different policy-makers have rapidly taken into account these recommendations and 44 

some governments have introduced tax on sugary drinks, among other measures 45 

developed to decrease the intake of free sugars (Briggs, 2016). In this scenario, it has been 46 

recently reported that the reformulation to reduce sugar concentration in sweetened 47 

beverages could be the most beneficial and healthy industry strategy (Briggs et al., 2017). 48 

Therefore, the use of high-potency sweeteners (also known as non-nutritive sweeteners 49 

or low-calorie sweeteners) and/or their blending with sugars is recognized as a 50 

technologically feasible, economically viable and effective strategy in reducing free 51 

sugars in foodstuffs (Gibson et al., 2017a; Di Monaco, Miele, Cabisidan, & Cavella, 52 

2018). The current high-intensity sweeteners (HIS) more commonly used in Europe are 53 

synthetic, such as aspartame, saccharin, sucralose, acesulfame-K, neotame, although 54 

some of them are derived from a natural source as is the case of steviol glycosides. 55 

However, due to the absence of solid scientific evidence supporting the role of synthetic 56 

sweeteners in preventing weight gain, together with the lack of studies on other long-term 57 

effects on health, the use of common synthetic sweeteners as part of a healthy diet is 58 

currently under question (Edwards, Rossi, Corpe, Butterworth, & Ellis, 2016; Borges et 59 

4  

al., 2017; Azad et al., 2017). In addition, HIS tend to have a different sweetness profile 60 

to natural sugars, often having a lingering sweetness and, in some cases, additional off-61 

notes such as bitterness or specific flavours such as liquorice (Prakash, Dubois, Clos, 62 

Wilkens, & Fosdick, 2008). In this context, it has been claimed that the replacement of 63 

free sugars with any HIS will continue to be primarily governed by the required sweetness 64 

profile, making sensory science and in-depth understanding of consumer attitude key 65 

players on the potential incorporation of any new sweetener into a normal diet (Miele et 66 

al., 2017). 67 

Carbohydrates with prebiotic properties, which are selectively utilized by host 68 

microorganisms conferring health benefit(s)  to the gastrointestinal tract (GIT), among 69 

other body sites (Gibson et al., 2017b), exhibit a high resistance to digestion and 70 

absorption in the upper GIT having, thus, a low calorific content. Prebiotic carbohydrates 71 

are mainly produced either by extraction from natural sources, as well as by enzymatic 72 

hydrolysis or synthesis using naturally-occurring polysaccharides or disaccharides (such 73 

as lactose, sucrose and maltose) (Díez-Municio, Herrero, Olano, & Moreno, 2014). The 74 

assessment of the sweetness properties of oligosaccharides with prebiotic properties, or 75 

with slow digestion rate, produced from natural sources and "green technology" can 76 

provide valuable insights to better understand their potential as suitable and healthy low-77 

calorie sweeteners. Although there are several studies dealing with the determination of 78 

the sweetness of carbohydrates, such as those evaluating monosaccharides (Schaafsma, 79 

2002; Gwak, Chung, Kim, & Lim, 2012), maltodextrins (Marchal, Beeftink, & Tramper, 80 

1999; Pullicin, Penner, & Lim, 2017), lactose (Pangborn & Gee, 1961), glucose polymers 81 

(Lapis, Penner, & Lim, 2014) or polyalcohols (Gwak et al., 2012; Grembecka, 2015), the 82 

information gathered on commercial prebiotic carbohydrates, such as lactulose, FOS, 83 

GOS or XOS, is scarce (Parrish, Talley, Ross, Clark, & Phillips, 1979; Niness, 1999; 84 

5  

Schaafsma, 2008; Bali, Panesar, Bera & Panesar, 2015; Samanta et al., 2015). 85 

Interestingly, recent works have demonstrated that the incorporation of GOS (Belsito et 86 

al., 2017) or XOS (Ferrao et al., 2018) into processed cheese led to an improvement of 87 

the sensory characteristics. 88 

In recent years, the effective production of a series of novel prebiotic oligosaccharides 89 

enzymatically synthesized, using microbial transglycosidases acting on sucrose, has been 90 

reported (Diez-Municio, Kolida, Herrero, Rastall, & Moreno, 2016a), and whose 91 

sweetness potential is unknown. Thus, the objective of this work is to comparatively 92 

evaluate the sweetness and flavour profiles of fourteen different carbohydrates, including 93 

novel prebiotics as well as a range of commercially available carbohydrates in order to 94 

infer findings from the relationship between the structural features and the sweetness 95 

properties of the tested carbohydrates. 96 

97 

2. Material and methods 98 

2.1. Carbohydrates and chemicals 99 

Orafti® HP, Orafti® P95 and Palatinose® were acquired from Beneo-Orafti 100 

(Tienen, Belgium) and IMO Syrup (isomaltooligosaccharide) was bought from Vitafiber 101 

(Bioneutra, Alberta, Canada). Kojibiose, leucrose, maltulose and turanose were acquired 102 

from Carbosynth (Compton, UK). Lactose and lactulose were purchased from Sigma-103 

Aldrich (Steinheim, Germany). All material was stored at ambient temperature, except 104 

for IMO Syrup which was stored at 5 °C. 105 

Water (Harrogate Spa mineral water) and white granulated sugar (Sainsburys, 106 

London, UK) used for sensory testing were purchased in local supermarkets in Reading 107 

(UK). 108 

109 

6  

2.2.Synthesis and purification of novel oligosaccharides 110 

The novel carbohydrates were produced by enzymatic synthesis using microbial 111 

transglycosidases acting on sucrose. 4-Galactosyl kojibiose (β-D-Gal-(1→4)-D-Glc-112 

(2→1)-α-D-Glc) was produced as described by Diez-Municio et al. (2012a), 113 

lactulosucrose (-D-Gal-(1→4)--D-Fru-(2→1)--D-Glc) as in Diez-Municio, Herrero, 114 

Jimeno, Olano & Moreno (2012b), lactosyl-oligofructosides (LFOS) (β-D-Gal-(1→4)-α-115 

D-Glc-[(1→2)-β-D-Fru]n, n = 2–4) as in Diez-Municio et al. (2015) and raffinosyl-116 

oligofructosides (RFOS) (α-D-Gal-(1→6)-α-D-Glc-[(1→2)-β-D-Fru]n, n = 2–5) as in 117 

Diez-Municio et al. (2016b). 118 

The synthesized carbohydrates were isolated and purified by high performance 119 

liquid chromatography with refractive index detector (HPLC-RID) from the 120 

corresponding reaction mixtures on an Agilent Technologies 1260 Infinity LC System 121 

(Boeblingen, Germany) using a Zorbax NH2 PrepHT preparative column (250 mm x 21.2 122 

mm, 7 µm particle size) (Agilent Technologies, Madrid, Spain). Two mL of reaction 123 

mixtures (approx. 150 mg of total carbohydrates) were eluted with acetonitrile:milli-Q® 124 

ultrapure water with a resistivity of 18.2 MΩ·cm at 25 °C (75:25, v:v) as the mobile phase 125 

at a flow rate of 21 mL/min for 30 min. The separated compounds were collected using 126 

an Agilent Technologies 1260 Infinity preparative-scale fraction collector (Boeblingen, 127 

Germany) and the fractions were evaporated in a rotatory evaporator R-200 (Büchi, 128 

Flawil, Switzerland) at a temperature below 25 ºC and freeze-dried to avoid any cross 129 

contamination (microbial or chemical). 130 

The obtained purified oligosaccharides were sterilized by filtration (0.22 μm 131 

filter). Moreover, in order to ensure all solvent was removed, total carbon, hydrogen, 132 

nitrogen and sulfur contents were determined in all the carbohydrates using a LECO 133 

7  

analyzer (Model CHNS-932, Leco Corp., St Joseph, MI) from the Service 134 

Interdepartmental Research (SIdI-UAM) in Madrid. 135 

All samples underwent microbiological clearance testing. The presence of yeast 136 

and molds, total and sporulated aerobic microorganisms and enterobacteria were analyzed 137 

in the samples. Serial dilutions were performed in triplicate with peptone water (Biocult 138 

BV, Roelofarendsveen, The Netherlands). Yeast and molds were plated on Sabouraud 139 

chloramphenicol agar and incubated at 25 ºC for 5 days. The total and sporulated aerobic 140 

bacteria were determined by plating appropriately diluted samples onto plate count agar. 141 

The samples were incubated at 30 ºC for 72 h for total aerobic bacteria and at 37 ºC for 142 

48 h for sporulated aerobic bacteria after heat treatment of stock dilution at 80 ºC for 10 143 

min. For enterobacteria counts, violet red bile dextrose agar was used and incubation was 144 

carried out at 30ºC for 24h. All microbial counts were reported as colony forming units 145 

per gram (cfu g-1). All culture media were of Difco (Becton, Dickinson & Company, 146 

Franklin Lakes, NJ, USA). 147 

148 

2.3.Conditions for sensory analysis 149 

The sweetness intensity of commercial and novel prebiotic oligosaccharides was 150 

evaluated using an experienced sensory evaluation panel of subjects. The study was given 151 

approval by the University of Reading Research Ethics Committee (UREC study number 152 

16_19). Sensory analysis was performed in an air-conditioned (23-24°C, room 153 

temperature) sensory laboratory with individual booths and artificial daylight. 154 

The sweetness intensity and several flavor attributes of novel and commercial 155 

oligosaccharides was carried out by a screened and trained sensory panel which consisted 156 

of 10 panelists (9 female, 1 male; 30-60 years of age) with between 5 months and 8 years’ 157 

experience. 158 

8  

The panelists were trained at the Sensory Science Centre (Department of Food 159 

and Nutritional Sciences, University of Reading, UK). Using a QDA (quantitative 160 

descriptive analysis) profiling approach the panel first developed a consensus vocabulary 161 

and then scored independently each attribute, in duplicate. 162 

The panel used 11 attributes to define the oligosaccharide samples (sweet, overall 163 

strength of off taste/flavor, bitter, cardboard/stale, candyfloss, sour, metallic, salty, crusty 164 

bread, perfume flavour and sweet aftertaste) as defined in Table 1. 165 

The training focused on ensuring each panelist could reliably score sweetness 166 

relative to four sucrose standards (5, 10, 20 and 26 g/L). The average panel ratings for 167 

these standards were 10, 35, 75 and 100 respectively on a 0-100 line scale, and hence 168 

these four positions were used as anchors to provide a structured scale on which to rate 169 

all oligosaccharide samples. All other attributes were scored as relative values using 170 

unstructured line scales (0-100). Due to the limited sample availability each panelist was 171 

presented with only 0.5 ml of sample for each scoring session. Therefore, training 172 

additionally focused on ensuring panelists were able to sip this small sample volume from 173 

a 30 ml transparent polystyrene cup and allow it to flow over the top of their tongue 174 

before swallowing and scoring sweetness reproducible. Palate cleansing before and 175 

between sample scoring was done using filtered water and low salt crackers (Carr’s water 176 

crackers, United Biscuits Ltd., Hayes, UK). 177 

Oligosaccharide samples were prepared as a 50 g/L solution (weighed to an 178 

accuracy of ±0.005 g) in mineral water (Harrogate Spa mineral water), stirring over a 179 

magnetic plate to ensure thorough sample dispersion. In a pilot tasting session it was first 180 

ensured that 50 g/L was of sufficient concentration to be tasted by all panel members 181 

(data not shown); a higher concentration was not used due to limited sample availability. 182 

The samples dispersed well and solubilized easily in water, with the exception of 183 

9  

raffinosyl-oligofructosides (RFOS) which were more difficult to disperse and separated 184 

out of solution on standing. However, all samples were shaken immediately before 185 

serving to each sensory panelist. Samples were labelled with random 3-digit codes and 186 

sample order presentation was done in a monadic sequential manner. 187 

The sucrose standards were presented at the start of each panel rating session for re-188 

familiarization in order that the panelists could score the sweetness of the 189 

oligosaccharides accurately against the standard anchors. 190 

The mean sweet ratings of the four sucrose standards were used to plot a dose-191 

response curve, the linear regression for which was Perceived Sweetness = 37.5 x Sucrose 192 

Concentration (g/L) (r2 = 0.98). The mean sweet ratings for each 50 g/L oligosaccharide 193 

were the converted to equivalent sweetness (ES) values from this equation. Sugars and 194 

sweeteners are usually compared to sucrose by relative sweetness (RS) values, the ES on 195 

a dry weight basis. To account for the 50 g/L of each oligosaccharide, the RS was 196 

determined as RS = ES /50. 197 

198 

2.4.Statistical analysis. 199 

Data were analyzed using a mixed model ANOVA where panelists were treated as 200 

random effects and samples as fixed effects, the main effects were tested against the 201 

sample by assessor interaction. Multiple pairwise comparisons were carried out using 202 

Fishers LSD and a significant difference was declared at an alpha risk of 5% (p 0.05). 203 

Data analysis was carried out using Senpaq software (Qi Statistics, Reading, UK). 204 

PCA tests and Spearman rank correlation analyses were done using the statistical 205 

software XLSTAT (Addinsoft, version 2015, Paris, France). 206 

207 

3. Results and Discussion 208 

10  

3.1. Chemical structure and degree of purity of the tested carbohydrates 209 

Table 2 shows the chemical structures and degree of purity of the carbohydrates 210 

used in the present study. A range of sucrose isomers sharing the monomer composition 211 

but differing in the glycosidic bond (leucrose, maltulose, turanose and palatinose) were 212 

included in order to determine the potential influence on the glycosidic linkage on the 213 

resulting sweetness. In addition, lactose and lactulose were assayed as disaccharides 214 

forming the core structure of some of the novel prebiotics tested. Moreover, a wide range 215 

of degrees of polymerization (DP) were studied (from 2 to an average of 23). Glycosidic 216 

linkages varied in the structures (α(1→2), β(1→4) and β(2→1) bonds), and the 217 

monomeric composition was based on glucose, galactose or fructose, which are the main 218 

building blocks of the majority of food oligosaccharides presently available or in 219 

development as functional food ingredients. For instance, kojibiose consists of two 220 

glucose units linked by an α(1→2) bond, whereas 4-galactosyl-kojibiose contains three 221 

monomers (two glucose and one galactose units), and lactulosucrose has three different 222 

monomers (galactose, fructose and glucose) linked by (1→4) and (2→1) bonds. In the 223 

case of RFOS, which contained galactose, glucose and up to five molecules of fructose, 224 

are linked by (1→6) and (2→1), respectively. Compounds having a higher DP, such 225 

as commercial oligofructoses, isomaltooligosaccharides and long chain inulin were also 226 

included. 227 

The degree of purity was determined in all assayed carbohydrates in order to avoid 228 

any bias in the sweetness properties induced by the possible presence of minor 229 

carbohydrates, especially monosaccharides. The levels of purity were satisfactory and 230 

ranged from 87 to 99% (Table 2). 231 

Microbiological assays showed that the microbial load (yeast and molds, total and 232 

sporulated aerobic bacteria, enterobacteria) was, in all assayed carbohydrates, lower than 233 

11  

103 cfu g-1, indicating that the synthesized oligosaccharides were microbiologically safe 234 

and could be used as a food ingredient. 235 

Determination of carbon, hydrogen, nitrogen and sulfur in the novel oligosaccharides 236 

(i.e., 4-galactosyl-kojibiose, lactulosucrose, RFOS and LFOS) revealed normal values for 237 

these elements, including low nitrogen contents (between 3 and 5.7 g/L) which is in 238 

accordance with their high degree of purity. 239 

240 

3.2. Sensory profile of commercial and novel carbohydrates 241 

Significant differences in sweet taste were found for the oligosaccharides tested 242 

with mean scores ranging from 11.2 to 68.3 (out of 100) (Table 3). Turanose was 243 

significantly sweeter than all other samples except leucrose. The sweet scores for 244 

disaccharides ranged from 49.8 to 63.0. Kojibiose was the disaccharide with the lowest 245 

sweet mean value (49.8), it was significantly less sweet than both leucrose and turanose 246 

and was not significantly different from either of the trisaccharides, lactulosucrose (46.5) 247 

and 4-galactosyl-kojibiose (41.4). Among the oligosaccharides having a DP above 3, the 248 

sweetest samples were the oligofructose with relatively low DP (Orafti® P95), and 249 

RFOS, followed by LFOS and IMO syrup, whereas the long chain inulin (Orafti® HP) 250 

was noticeably the least sweet sample. Differences in sweet aftertaste (post swallowing) 251 

followed the same trend (Table 3). The relative sweetness (RS) of the oligosaccharides 252 

varied from 0.06 to 0.36, indicating that on a weight basis these molecules had between 253 

6% and 36% the sweetness of sucrose. 254 

The differences in the overall strength of off taste/flavours in the oligosaccharide 255 

samples were also significant with palatinose having the least off flavor value (8.9), and 256 

kojibiose having a significantly higher level than all other assayed carbohydrates. 257 

Although bitter taste and cardboard/stale flavor values were particularly low in all 258 

12  

carbohydrates, values determined for lactosyl-oligofructosides (LFOS) were significantly 259 

higher than in the rest of the studied carbohydrates. None of the remaining off-notes 260 

characterized were significantly different between samples. There was a candyfloss 261 

(cooked sugar) flavor at low levels in some samples, particularly in leucrose and 262 

maltulose that was absent in LFOS. Crusty bread flavor tended to be slightly higher in the 263 

commercial oligosaccharides (DP≥3), specifically in oligofructose (Orafti P95®), while 264 

the novel oligosaccharides did not present this attribute. Kojibiose and 4-galactosyl-265 

kojibiose were rated slightly higher for the perfume note, although at a low level with no 266 

significant differences between samples. Lastly, sour (rancid), salty and metallic did not 267 

appear to substantially contribute to the overall off flavour, nor to discriminate between 268 

samples. 269 

In order to better correlate sweetness and chemical structure of the tested 270 

carbohydrates, a multivariate analysis was carried out with the aim to group the different 271 

carbohydrates and visualize main trends. Concretely, Figure 1 graphically shows the 272 

Principal Component Analysis (PCA) of the sweet scores and DP, using two other factors 273 

regressed onto the plot as supplementary variables (presence of ketose groups and types 274 

of linkage). The main factor contributing to sweetness was a low DP, which is in good 275 

agreement with previous findings (Kaulpiboon, Rudeekulthamrong, Watanasatitarpa, Ito 276 

& Pongsawasdi, 2015). As can be seen in Figure 1 the DP and mean sweet score are at 277 

opposite sides of dimension 1, the Spearman’s correlation coefficient between the two 278 

was -0.87 (p<0.0001) as indeed all oligosaccharides with a DP above 3 were substantially 279 

less sweet. Moreover, presence of a ketose sugar moiety did not have a significant 280 

influence on sweetness although there was a very weak non-significant trend that the 281 

presence of a ketose sugar led to higher sweetness (Spearman’s correlation coefficient 282 

0.17, p=0.56). This weak trend could partly explain the fact that kojibiose, the only tested 283 

13  

disaccharide comprised by two glucose monomers, had a lower sweetness than the 284 

sucrose isomers or lactulose (comprising galactose and fructose monomers). In this sense, 285 

Schaafsma (2002) stated that fructose has higher sweetness properties than glucose. 286 

Moreover, despite turanose (α-D-Glc-(1→3)-β-D-Fru) was significantly sweeter than 287 

maltulose (α-D-Glc-(1→4)-β-D-Fru) and palatinose (α-D-Glc-(1→6)-β-D-Fru) but not 288 

significantly sweeter than leucrose (α-D-Glc-(1→5)-β-D-Fru), PCA revealed the lack of a 289 

clear relationship between the type of linkage and sweetness of the resulting 290 

oligosaccharide (Figure 1). 291 

Concerning the novel prebiotics tested in this study, the purified trisaccharides 292 

lactulosucrose and 4-galactosyl-kojibiose had around 25% of the sweetness of sucrose 293 

which was similar to or even higher than the relative sweetness of oligofructose with low 294 

DP (Orafti® P95) (Table 3) which has previously been described as sweet with a pleasant 295 

flavor and, consequently, it could be used in combination with high intensity sweeteners 296 

to replace sucrose, providing a well-balanced sweetness profile (Niness, 1999). LFOS 297 

(DP 4-6) and RFOS (DP 4-7), which exhibited around 18% of the sweetness of sucrose, 298 

were significantly sweeter than the long-chain inulin (Table 3). 299 

Finally, the tested oligosaccharides were also characterized by other flavour 300 

attributes, however, the results showed no clear association regarding off notes related to 301 

commercial or non-commercial oligosaccharides. 302 

303 

4. Conclusions 304 

Information related to sweetness and sensory properties of prebiotic 305 

oligosaccharides which could potentially be used as sweeteners is rather scarce at present. 306 

The present study may initially contribute to fill this gap because a wide range of 307 

commercial and novel prebiotic oligosaccharides displaying different chemical 308 

14  

structures, such as degree of polymerization, monomer composition and order, presence 309 

of ketone vs. aldehyde group, or different glycosidic linkages, were assayed in order to 310 

propose relationships between carbohydrate structure and sweetness properties. Data 311 

from the sensory panel and further supported by PCA pointed out that chain length was 312 

the most relevant factor in determining the sweetness potential of a carbohydrate. Thus, 313 

disaccharides had higher sweetness values (49.8-68.3) than trisaccharides (41.4-46.5) 314 

which, in turn, exhibited greater sweetness than mixtures of oligosaccharides having DP 315 

above 3 (11.2-37.1). Less remarkably, a weak and non-significant trend indicated that the 316 

presence of a ketose sugar moiety led to higher sweetness, whereas the type of glycosidic 317 

linkage did not have a clear impact on the sweetness properties of the tested 318 

oligosaccharides. 319 

The novel prebiotic oligosaccharides studied in the current study had between 18 320 

and 25% of the sweetness of sucrose (relative sweetness), showing, thus, a sweetness 321 

potential in line with other commercial prebiotics. Therefore, these findings suggest a 322 

potential use for clean tasting prebiotics as partial sugar replacers, or in combination with 323 

high intensity sweeteners, to provide a well-balanced sweetness profile. 324 

325 

Acknowledgments 326 

This work has been funded by Optibiotix Health plc (York, UK) and by Ministerio de 327 

Economía, Industria y Competitividad (MINEICO) of Spain (project AGL2017-84614-328 

C2-1-R). L. R-A. thanks the Spanish Research Council (CSIC) and the Spanish Ministry 329 

of Economy and Competitiveness for a “Juan de la Cierva-Formación” contract. 330 

331 

15  

Declarations of interest: The authors declare the following competing financial 332 

interest(s): SK is the Research and Development director of Optibiotix Health plc. 333 

16  

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Figure captions 468 

Figure 1. Principal component analysis (PCA) plot of the sweet scores and degree of 469 

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Additional supplementary variables as presence of ketose groups and type of linkage were 471 

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Observations plot. 473