Application of HPLC Coupled with a Charged Aerosol Detector to the Evaluation of Fructose, Glucose, Sucrose, and Inositol Levels in Fruit Juices, Energy Drinks, Sports Drinks, and Soft Drinks
1
Department of Bromatology, Medical University of Gdansk, Al. Gen. J. Hallera 107, 80-416 Gdansk, Poland
2
The Institute of Sport and Health Studies, State University of Applied Sciences in Koszalin, ul. Leśna 1, 75-582 Koszalin, Poland
*
Author to whom correspondence should be addressed.
Beverages 2024, 10(4), 94; https://doi.org/10.3390/beverages10040094
Submission received: 16 July 2024
/
Revised: 26 September 2024
/
Accepted: 29 September 2024
/
Published: 1 October 2024
(This article belongs to the Section Tea, Coffee, Water, and Other Non-Alcoholic Beverages)
Abstract
:The study aimed to estimate the levels of fructose, glucose, sucrose, and inositol levels in sweetened beverages with a newly developed method using HPLC coupled with a charged aerosol detector (CAD). In total, 85 commercially available non-alcoholic beverages, including 18 energy drinks, 8 sports drinks, 15 soft drinks, 14 fruit drinks, 7 fruit nectars, and 22 fruit juices were analyzed by HPLC-CAD. The method was validated, and it was characterized by a wide concentration range (1–150 µg/mL), sensitivity, and good accuracy (94.9–103%). The results showed significant variation in fructose, glucose, and sucrose concentrations in energy drinks, sports drinks, soft drinks, fruit drinks, fruit nectars, and juice. The highest total sugar contents (fructose, glucose, sucrose, and inositol) were found in energy drinks (14.2 g/100 mL), followed by fruit nectars (13.7 g/100 mL) and soft drinks (12.7 g/100 mL). Statistical analysis (Spearman correlation test, Kruskal–Wallis test) of the data showed significant relationships between particular sugars in the analyzed products.
1. Introduction
The metabolic response is identical, regardless of the sugars’ origin, whether natural or added [1]. However, natural carbohydrates are usually accompanied by essential nutrients and minerals, whereas those added only supply calories without extra nutritional value. The term “added sugars” refers to sucrose, fructose, glucose, starch hydrolysates (glucose syrup, high-fructose syrup), and other isolated sugar preparations that are used as such or added during food preparation and manufacture [2,3]. Overconsumption of both naturally occurring and added sugars, especially in the form of sweetened beverages, increases the risk of developing dental cavities and becoming overweight [1,2,3,4,5,6].
Dietitians and researchers conducting dietary surveillance should be able to estimate accurately the consumption of sugars, such as glucose, fructose, and sucrose. High-quality and nutrient dense diets have been demonstrated to have a positive correlation with food labeling [5]. It would be highly beneficial to change food labels to include the amounts of added sugars. However, even national food composition tables contain limited data on mono- and disaccharides [7,8,9,10].
Therefore, there is a need to evaluate the composition of sweetened beverages in terms of fructose, glucose, and sucrose levels using reliable analytical approaches. Various methods have been reported for the determination of sugars in food matrices. The first and most widely used methods concern high-performance liquid chromatography coupled with a refractive index (RI) detector [11,12,13,14] but also methods with electrochemical detectors can be found [15]. There are also applications that are based on gas chromatography [16,17,18,19] or capillary electrophoresis (CE) [20]. Gas chromatography, however, is not commonly utilized because it requires time-consuming and laborious sample derivatization. Because HPLC procedures only require basic water dilutions, they are the preferable option.
According to the Official Methods of Analysis of AOAC International [13], a recommended method developed for the analysis of major saccharides in corn syrup (AOAC 979.23) is based on an HPLC system coupled with an RI detector. A differential refractive index detector detects peaks based on the difference in the refractive index between the analyte and the background mobile phase. It is one of the universal detectors for the analysis of substances that has refractive index. The advantage of the RI detector is its ability to work with relatively high concentrations of eluents and to cover the entire refractive index range from 1.000 to 1.750 RI with a single, easily balanced cell [21]. However, it also has some disadvantages, such as low sensitivity compared to UV or other detectors and a limited ability to distinguish the sample ions from the mobile phase ions. Moreover, it is sensitive to factors that affect refractive index, such as temperature, pressure, and mobile-phase composition [21]. Therefore, the refractive index detector is not suitable for use with gradient elution. UV detection, which is one of the most popular methods in analytical chemistry, cannot be used for carbohydrate quantification as the compounds do not possess chromophores. However, mass spectrometry and fluorescence detection can be applied, but they require a lengthy and time-consuming derivatization process and are quite expensive.
Other universal detectors that can be used for carbohydrate analysis are based on evaporative light scattering detection (ELSD) or charged-aerosol detection (CAD). Both detectors are based on a common principle, i.e., pneumatic nebulization of the mobile phase containing the analyte, which elutes from the column, to form droplets that are subsequently dried into particles. They differ in the mode of detection; ELSD uses a laser beam and measures the reflected light scattered to a sensitive photomultiplier, whereas CAD uses a high-voltage corona needle to charge nitrogen gas, which collides with analyte particles, resulting in the formation of charged particles, which are collected by a sensitive electrometer [22]. However, the ELSD detector is claimed to be less sensitive and not very precise in comparison with the CAD detector [22,23,24,25]. Moreover, ELSD response curves are typically more complex and less consistent among the analytes than those of CAD and are often sigmoidal. Among all universal detectors, CAD works independently of the compound chemical structure. It is also easy to operate, measures the physical properties of the analyte, and responds to almost all non-volatile species.
The study aimed to develop a new method for sugar determination using HPLC-CAD and to determine the composition of sweetened beverages concerning the fructose, glucose, sucrose, and inositol levels.
2. Materials and Methods
2.1. Apparatus and HPLC Conditions
The equipment consisted of an HPLC Ultimate 3000 system (Dionex, Germering, Germany) and CAD detector (ESA, Chelmsford, MA, USA). Data processing was carried out with Chromeleon 6.8 software (Dionex). The nitrogen gas (nitrogen generator Sirocco-5, Schmidlin-DBS, Geneva, Switzerland) flow rate was regulated automatically at 35 psi and monitored by the CAD device. The CAD detector response range was set to 50 pA on a full scale, and a medium filter was applied. The separation column was a Shodex Asahipak, NH2P-50 4E 5 µm (4.6 × 250 mm), and the column temperature was set at 25 °C. Satisfactory chromatographic separation was obtained using a mobile phase composed of water/acetonitrile in a ratio of 25/75 (v/v) with an isocratic run at a flow rate of 0.8 mL/min. The injection volume was 10 µL.
2.2. Chemicals, Reagents and Standards
Sucrose, glucose, fructose, inositol, and erythritol were purchased from Sigma Aldrich. Other chemicals were analytical-reagent grade, and all aqueous solutions were prepared using ultra-pure water (18.2 MΩ/cm) from a Milli-Q system (Millipore, Burlington, MA, USA). HPLC-grade acetonitrile was obtained from J.T. Baker (Malinckrodt Baker B.V., Deventer, Holland). Standards were made by dissolving 0.1000 g of each individual analyte in ultra-pure water in separate 100.00 mL volumetric flasks and were further successively diluted with ultra-pure water. The standard working solutions (1–150 µg/mL) were prepared daily by appropriate dilution from the individual stocks. All solutions were stored in a refrigerator (+4 °C) when not in use. Samples were stored for no more than 14 days. The chromatogram of the analyzed standards is shown in Figure 1.
2.3. Sample Preparation
Eighty-five commercially available non-alcoholic beverages, including 19 energy drinks, 8 sports drinks, 15 soft drinks, 14 fruit drinks, 7 fruit nectars, and 22 fruit juices, were randomly obtained from the local market in Gdansk (Poland), although some of them are sold worldwide. The characteristics of the analyzed products are listed in Table A1. The abbreviation ED stands for energy drinks, SD for sports drinks, and SfD for soft drinks. Numbers (1–9) in the upper index correspond to various producers. Capital numbers (1–17) correspond to the particular products. There were 3 samples taken for each collected type of beverage from different batches. Then, after two weeks, most of the products were duplicated in order to confirm the first results. Each sample represented unique production batches as well as production time. In general, the purchased products represented a variety of beverages that contained various levels of natural and added sugars. The analyzed beverages were stored in a dark, cool place for no more than 14 days. All of them were originally packaged and sealed. The analyzed products were still suitable for consumption at the time of analysis, as they had not expired. After opening, samples were immediately prepared for chromatographic analysis. Carbonated drink samples were degassed in a laboratory shaker bath (Elpan type 357, Elpan, Warsaw, Poland) for 30 min (at room temperature, 100 rpm) and in an ultrasound bath (VWR USC900THD, Kuala Lumpur, Malaysia) for 15 min, and filtered through filter paper MN 615¼ (Ø 110 mm; Macherey-Nagel, Dűren, Germany). Then, samples were diluted with ultra-pure water as required, and prior to analysis, all the final solutions were filtered through 0.45 µm syringe filters (regenerated cellulose, RC-45/25; 0.45 μm pores; Macherey-Nagel, Dűren, Germany). Samples were stored in a refrigerator (+4 °C) when not in use. All products were stored under the same environmental conditions and then prepared in the same manner. Each of the 510 analytical samples was injected in triplicate.
2.4. Validation of the Method
The calibration curves were obtained by plotting concentration (µg/mL) against peak area. For each compound analyzed, 12 standards were prepared in the range of 1–150 µg/mL; each one was injected six times.
The limit of detection (LOD) and the limit of quantification (LOQ) were calculated based on the calibration curve, using formulas proposed by Konieczka and Namieśnik [26], i.e., 3 SD/a and 10 SD/a (SD is the standard deviation of the curve and “a” is the slope of the curve).
Erythritol was chosen as an internal standard, which was helpful for monitoring retention time and the area under the peaks. The concentration of the internal standard throughout the analyses was 25 µg/mL. The recovery test was used to evaluate the accuracy of this method. The selected samples of orange juice and energy drink were analyzed before and after the addition of known quantities of erythritol, fructose, glucose, sucrose, and inositol in the amounts of 10, 25, and 50 µg/mL and 15, 30, and 45 µg/mL, respectively. The average recoveries were calculated using the following formula: recovery (%) = (observed amount-original amount)/spiked amount × 100% [26].
The precision of the method (expressed as RSD) was calculated by measuring the repeatability and reproducibility of each sugar in the samples. The inter- and intra-day experiments were evaluated for samples of orange juice with the addition of known quantities of erythritol, fructose, glucose, sucrose, and inositol in amounts of 10, 25, and 50 µg/mL. The precisions were calculated from three consecutive injections for each concentration. Intermediate precision was calculated over a period of 2 days.
2.5. Statistical Analysis of Data
The results were tested for normality using the Shapiro–Wilk and Kolmogorov–Smirnov tests. The variables did not meet the criteria of normality; thus, non-parametric tests were applied. The results were subjected to Kruskal–Wallis and Dunn’s tests as well as R-Spearman correlation. Statistical analysis of the data was performed using Statistica 13.0 (TIBCO Software Inc., Palo Alto, CA, USA).
3. Results
3.1. Results of the Validation of the Method
The developed method was fully validated by evaluating the linearity range, intra- and inter-day precision and repeatability, limit of quantification (LOQ), limit of detection (LOD), and accuracy. The data concerning the method validation are summarized in Table 1.
All calibration curves showed good linear regression (R2 = 0.9995–0.9999) across the entire range. The linearity was maintained up to 150 µg/mL for all the analyzed samples and met the requirements typically specified in method validation protocols. The observed precision (expressed as RSD) ranged from 1.02% to 2.79%, whereas recoveries (as a measure of accuracy) were between 94.9% and 102% for the orange juice and between 96.0% and 105% for the energy drink (Table 1). The limit of detection and LOQ for the four analytes were in the range of 0.05–0.16 µg/mL and 0.16–0.48 µg/mL, respectively. The values obtained for each substance are shown in Table 1.
3.2. Quantitative Analysis of Sugars in Beverages
The developed HPLC-CAD method was successfully applied to the analysis of fructose, glucose, and sucrose in energy drinks, sports drinks, soft drinks, fruit drinks, fruit nectars, and juices. Inositol was not detected in the analyzed samples. Table 2 summarizes the composition of the analyzed sugars in sweetened beverages, whereas Table A2 provides full details concerning the sugar content in the analyzed samples.
The results showed significant variation in fructose, glucose, and sucrose concentrations within each group analyzed. The highest total sugar contents were found in energy drinks (14.2 g/100 mL), followed by fruit nectars (13.7 g/100 mL) and soft drinks (12.7 g/100 mL). The highest fructose and glucose contents were determined in soft drinks (5.77 and 6.04 g/100 mL, respectively), whereas the lowest were in sports drinks (1.02 and 2.63 g/100 mL, respectively). In the case of sucrose, its highest levels corresponded to samples of fruit nectars (5.56 g/100 mL), and the lowest to soft drinks (1.13 g/100 mL) (Table 2).
Among energy drinks, the highest total sugar contents were determined in beverages ED8 and ED6, i.e., 17.2 g and 16.8 g/100 mL, respectively (Table A2). The first energy drink was also characterized by the highest glucose (8.10 g/100 mL) and fructose (7.32 g/100 mL) levels. Although several energy drinks, including ED7 and ED8, did not list the addition of HFCS on their labels (Table A1), they contained nearly equal amounts of glucose and fructose, which could be attributed to sucrose degradation over time due to pH changes.
On average, sports drinks contained 1.02 g of fructose per 100 mL, with the highest value observed in SD3, a beverage with added juice. In contrast, three sports drinks—SD1, SD2*, and SD5—did not contain any fructose (Table A2).
Fruit drinks, which typically contain 0 to 20% juice, had varied fructose concentrations, ranging from 0.16 g in lemon drink6 to 7.52 g/100 mL in apple drink3 (Table A2). Among the 14 fruit drinks tested, lemon drink6 contained the lowest amounts of monosaccharides but quite notable amounts of sucrose. This finding is in accordance with the producer’s declaration. Lemon drink5 had the highest amount of sucrose. The lowest concentration of this disaccharide was determined in apple drink3 (1.51 g/100 mL).
Variations in fructose, glucose, and sucrose levels were significant in fruit nectars and juices (Table 2). It might be explained by the fact that fruit nectars are obtained by adding water, sugar, and/or honey to fruit juice or fruit juice reconstituted from concentrate or their mixtures [26,27,28]. The amount of juice added is strictly determined by legislation [27,28,29] according to the type of nectar, and so the minimum level for banana, citric, or currant nectar is 25%, whereas for pineapple or apple it is 50%. The amount of added sugars in nectars cannot exceed 20% by weight of the finished product and can be partially replaced by intense sweeteners if reduced energy value is required [27]. The highest levels of fructose were determined in apple juices and black currant nectars (Table A2). In addition, black currant nectar4 did not contain sucrose but only fructose and glucose (8.48 g and 6.76 g/100 mL, respectively), which implies the use of HFCS during its production.
Among fruit juices, the highest total carbohydrate content was determined in pineapple juice6-19.36 g/100 mL. Apple juices were characterized by the highest fructose concentration (7.65–7.86 g/100 mL), whereas pineapple juice6 contained the greatest amounts of glucose (6.3 g/100 mL) (Table 3). Sucrose levels in different juices were varied and ranged from 1.28 g/100 mL (apple juice5) to 6.91 g/100 mL (pineapple juice6) (Table A2).
An exemplary chromatogram of the analyzed energy drinks is depicted in Figure 2.
According to FDA guidelines and those of the European Commission Health and Consumers Directorate, products can contain up to 120% of the stated sugar content [28,30]. In general, the total sugar content within each group varied from the information provided by the manufacturer, with some products containing more and others containing less sugar than the label stated (Figure 3). The total sugar content of the analyzed beverages ranged from 45.3% to 186% of that listed on the label (Figure 3). The consistency of labeling was confirmed for all sugar-free products. When comparing the sugar content declared by the manufacturer with our results, it can be concluded that most of the analyzed energy drinks contained more sugar than indicated on the packaging. In contrast to energy drinks, the differences between the sugar content of sports drinks declared by the manufacturer and the values found in the analysis are much smaller, ranging from +2% to +13%. This might be evidence of the producers’ awareness of the target group’s (athletes) needs and requirements. In all soft drinks (except for light beverages in which no sugars were detected) and most fruit drinks, an excess of carbohydrates compared to the declared value was found. Most fruit drinks were sweetened with both HFCS and sugar (Table A1). In the case of fruit nectars, the largest difference between the declared and found values, amounting to +59.8%, was recorded for one black currant nectar4. Interestingly, the label of this product listed both sugar and HFCS as components of the final product (Table A1). On the other hand, grapefruit1 (sweetened with HFCS) and banana nectars4 (sweetened with sucrose) were found to have lower sugar content (sum of fructose, glucose, and sucrose) than declared by the manufacturer (−12.35% and −2.91%, respectively).
3.3. Recommended Daily Requirements
All the results were recalculated to one serving portion (in Poland—one glass, i.e., 250 mL) to facilitate comparison between various sweetened beverages. On average, one glass of the analyzed beverages provides between 14 and 35.4 g of sugars (Table 3). Their highest amount was determined for energy drinks, most of which contained only added sugars such as sugar, sucrose, or HFCS (Table A1). The consumption of an average glass of energy drink provides 142 kcal, which constitutes 4.72% of the total daily energy intake for a Polish male adolescent (3000 kcal) [31]. This group was selected as a representative because, according to Polish studies [32,33], a significant portion of this population drinks sweetened beverages. Although fruit juices supply 3.90% of total daily energy, they also provide natural sugars, as they are produced from fruit concentrates (Table A1).
3.4. Statistical Analysis
The statistical analysis (Spearman correlation test) of the data showed a significant relationship between particular sugars in the analyzed products. For instance, fructose content was negatively correlated with sucrose (p < 0.001) and positively correlated with glucose (p < 0.001). There was also a strong negative relationship between glucose and sucrose content (p < 0.001). The presence of fructose typically indicates the use of HFCS in the product, which could explain the negative correlation. A negative correlation with sucrose can result from the addition of sugar to a beverage, which usually signifies the absence of HFCS.
The Kruskal–Wallis test identified relationships between types of beverages and specific sugar content. There was a strong influence of the type of drink on fructose (H = 24.8, p = 0.0002), glucose (H = 39.3, p = 0.0000), and sucrose levels (H = 24.6, p = 0.0002). Further analysis using the post hoc test confirmed significant differences (p < 0.05) in multiple comparisons. The following product groups showed notable differences in the averages of specific sugars: glucose in energy drinks versus sports drinks, fruit drinks, and juices; sports drinks versus soft drinks; soft drinks versus fruit drinks; fructose in sports drinks versus soft drinks and fruit juices; soft drinks versus fruit drinks; and sucrose in soft drinks versus fruit drinks and nectars.
4. Discussion
We developed a novel method using HPLC coupled with a CAD detector for the simultaneous analysis of fructose, glucose, sucrose, and inositol. This method is characterized by a comparable coefficient of determination (linearity), precision, and accuracy when compared to the method developed for the International Society of Beverage Technologists (ISBT) and verified by White et al. [34]. The cited method [34] used high-performance liquid chromatography coupled with an RI detector and was adapted for measurement of HFCS sugars (fructose, glucose, maltose, maltotriose, and maltotetraose) in carbonated beverages. For the main sugar of interest, fructose, clean separation and baseline resolution were obtained, while acceptable resolution was achieved for the other sugar components.
However, the CAD detector in both isocratic and gradient elution can also be used to determine other compounds with different physicochemical characteristics in the same samples, such as artificial sweeteners, preservatives, acidity regulators [35], or bioactive compounds like caffeine, which can be found in various beverages [36]. In our study, the total run time was 20 min to check for peaks from any non-analyzed sugars. Since no other peaks were observed, the analysis time could be shortened to 14 min, as all the analyzed sugars were separated within 12 min. This is faster than the methodology proposed by White et al. [34], where peaks were separated within 16 min.
Our data indicate discrepancies between the declared and determined values for the analyzed non-alcoholic beverages. Other researchers also found differences between the values declared and determined for the sweetened beverages [37]. Walker et al. [19] analyzed beverages sweetened with and without HFCS and found significant differences between the total sugar content declared by the producer and that determined in the laboratory. Similar results were obtained for commercial infant formulas, baby foods, and common grocery items marketed toward children [18].
The Polish Food Composition Tables [8] contain only limited data on total carbohydrates and sucrose content in beverages. When comparing the sucrose content in cola-type drinks, a significant difference was observed between the data in the tables [8] and our results. In our samples, the presence of glucose and fructose was demonstrated, with negligible sucrose content, which may indicate the addition of HFCS instead of sugar. However, the total sugar content was similar. This may suggest that the manufacturer previously added sucrose to this type of drink but has now replaced it with HFCS. In our studies, we determined different levels of total sugar content in orange juice (8.55–11.41 g/100 mL), depending on the manufacturer. Kunachowicz et al. [8] indicate that the total sugar content is 9.9 g/100 mL, while the sucrose content is 2.2 g/100 mL. We determined the sucrose content in orange juice to be in the range of 3.73–5.10 g/100 mL (Table A2). In the case of apple juice, the determined sucrose levels were comparable to those in the literature [8], while the total sugar content in our case was higher. Similarly, for pineapple and grapefruit juices, our results were higher than those reported in the Polish nutritional value tables [8]. According to Kunachowicz et al. [8], black currant nectar contains an average of 12.7 g of sugars per 100 mL, while the average sugar content in the same product, determined in the present study, amounted to 16.7 g/100 mL. The discrepancies between the declared and determined total sugar content in fruit nectars and juices can also be partially explained by the diversification of sugar profiles in fruits. In addition to fructose, often referred to as fruit sugar, glucose, sucrose, maltose, and various sugar alcohols, including sorbitol and xylitol, can also be found. Among fruits, there are significant differences in sugar profiles. For instance, the highest percentage of fructose is found in apples and pears (65% and 75%, respectively), whereas other fruits contain between 42% and 55% [38]. This variability can also be observed in our results. Apple juices, compared to others, contained the highest levels of fructose (Table A2). Menezes et al. [10] found that pineapple is a rich source of soluble sugars, which might explain their levels in the juice. According to Sanz et al. [39], freshly squeezed apple juice contains up to 13.3 g of sugars in 100 mL, which is higher than the values obtained in this study. Such disparities in sugar content might result from the varied composition of products, which depends on factors like origin, harvest time, and weather conditions in a given year.
Fruits also contain mono-, di-, and oligosaccharides, particularly fructo-oligosaccharides and raffinose-family oligosaccharides, as well as fiber, which is not classified as sugar [14]. Nevertheless, sugar alcohols and oligosaccharides do not constitute a major fraction of the total carbohydrates in fruit juices. They are rarely present in fruit juices, most likely as a result of technological processing [40]. Türkmen and Ekşi [41] determined the presence of sorbitol and xylitol in pomegranate juice in concentrations ranging from 51 to 200 mg/L. According to Jovanovic-Malinovska et al. [14], most fruits are characterized by low levels of fructo-oligosaccharides, with the highest value observed in nectarines (0.89 g/100 g fresh weight). However, the CAD detector used in this study enables the detection of all sugars present in the sample. Nevertheless, no peaks other than those corresponding to glucose, fructose, and sucrose were observed. Furthermore, according to White [34], HFCS-42 and HFCS-55, which are commonly used as sweeteners even in fruit nectars, are mixtures of fructose (42 or 55%) and glucose, with a small percentage of residual glucose oligosaccharides. However, in our method, we did not estimate the concentration of maltose, which could alter glucose and fructose estimates, or other oligosaccharides; thus, we cannot confirm the reasons for these discrepancies. Initially, we tried to determine the amount of maltose, which ought to be observed at 12.5 min under the given conditions (simultaneously with inositol). Unfortunately, we were unable to detect maltose in any of the samples (LOD = 0.32 µg/mL) (Figure 1). In the case of beverages sweetened with sucrose, the inversion of sugar in an acidic environment should also be considered. Sucrose, which is a disaccharide comprising glucose and fructose in equal parts, hydrolyzes at a low pH of beverages and other foods, releasing both monosaccharides. According to White [34], this process can result in a misleading sugar profile in the product, with equal parts of glucose and fructose, which may suggest the addition of HFCS.
The beverage market continues to grow, with more and more people buying sweetened, carbonated, and non-carbonated drinks [42]. In 2012, an EFSA study team [43] conducted a survey on the consumption of energy drinks in Europe. The survey found that 68% of respondents who are most likely to consume energy drinks are adolescents, followed by adults and children [43]. In Poland, the consumption of soft drinks varied between 2015 and 2021. In 2015, each person consumed 99.9 L of soft drinks, according to Statista [44]. This quantity decreased to 91.4 L by 2021. The largest amount, 103.1 L, was consumed in 2019. Piekara and Krzywnos [33] showed that soft drink consumption declines as Polish consumers age. Drinks with added sugars are consumed less frequently by adults than by younger consumers and less commonly by women than men. Energy drinks are most commonly consumed by people between the ages of 30 and 44. The authors [33] speculate that it might be related to the respondents’ overall physical activity and profession. They also suggest that the consumption of sweetened beverages may be contributing to obesity in Poland, and therefore, these products should not be consumed in excess. The intake of sweetened beverages among Polish adolescents was found to exceed the recommendations of the World Health Organization (WHO) based on an analysis conducted by Kowalska et al. [32]. Drinking substantial volumes of sweetened beverages during the day resulted in the consumption of a significant, additional amount of energy by a large group of the surveyed adolescents. They estimated that the energy derived from consumed beverages amounted to an average of 256 kcal per capita per day, accounting for a mean of 8.7 ± 9% of daily energy requirements [32]. Considering the group of beverages in terms of energy provided, it was mainly derived from the group of fruit drinks. In our study, we found that fruit nectars, along with energy drinks, provided the greatest amounts of energy. The main source of energy in sweetened beverages is sugar. Campos-Ramirez et al. [45] suggested that the sugar content unique to caloric drinks is the addictive component capable of stimulating the dopaminergic and opioid systems. This effect may promote excessive sweetened beverage consumption.
Studies regarding beverage consumption rely on nutritional data provided on labels by the producer or in national food composition tables. Unfortunately, the latter usually contain scarce data concerning mono- and disaccharides. That is why it is important to constantly monitor the composition of beverages. Our data can greatly benefit such assessments, making them more accurate and reliable.
5. Conclusions
The developed HPLC-CAD method is fast, reliable, simple, and offers excellent linearity and good repeatability, making it appropriate for routine analysis in the food industry. Accurately quantifying specific sugars in sweetened beverages, which are sources of fructose, glucose, and sucrose in our diet, is important. Therefore, our research provided new data on the sugar content of sweetened beverages, which is significant for estimating their consumption. This research should be continued on a larger scale to ensure accurate estimates of sugars in the daily diet, which is paramount to our metabolic health.
Author Contributions
Conceptualization, M.G.; Data curation, M.G.; Formal analysis, M.G.; Funding acquisition, A.L.; Investigation, M.G.; Methodology, M.G.; Project administration, M.G.; Supervision, M.G. and P.S; Validation, M.G.; Visualization, M.G.; Writing—original draft, M.G.; Writing—review and editing, M.G. and P.S. All authors have read and agreed to the published version of the manuscript.
Funding
This work was funded by Polish National Scientific Research Fund, grant No. N N404 270840.
Data Availability Statement
All data are contained within the article.
Acknowledgments
I would like to thank Monika Mróz for her laboratory support.
Conflicts of Interest
The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.
Appendix A
Table A1.
Characteristics of the analyzed products.
| Product | Declared Sugar Content (g/100 mL) | Type of Sugar Listed on Label |
|---|---|---|
| Energy drinks | ||
| ED1 | 10.8 | sugar, HFCS |
| ED2 | 9.4 | sugar |
| ED3 | 10.8 | sugar |
| ED3 * | 0 | - |
| ED4 | 10.8 | sugar, HFCS |
| ED5 | 10.8 | sugar |
| ED6 | 11 | sucrose, glucose |
| ED7 | 13.3 | sugar |
| ED8 | 12.2 | sugar, vegetable juice (color), fruit juices (grapes, blackberry, black currant, strawberry, elderberry, raspberry) |
| ED9 | 10.6 | sugar, glucose |
| ED10 | 10.6 | sugar, glucose |
| ED11 | 12 | sugar, glucose syrup |
| ED12 | 11 | sucrose, glucose |
| ED13 | 13.5 | sugar, glucose |
| ED14 | 10.9 | sugar |
| ED14 * | 0 | - |
| ED15 | 3.8 | sugar, HFCS |
| ED16 | 11 | sugar |
| ED17 | 10.5 | sugar |
| Sports drinks | ||
| SD1 | 5.8 | glucose, maltodextrin |
| SD2 * | 0 | - |
| SD3 | 8.5 | concentrated apple juice, sugar, concentrated aronia juice |
| SD4 | 6 | sugar, dextrose |
| SD5 | 3.9 | glucose, maltodextrin |
| SD6 | 6.65 | sugar, glucose syrup |
| SD7 | 6.36 | HFCS |
| SD8 | 3.8 | dextrose, fructose |
| Soft drinks | ||
| SfD1 | 11.2 | HFCS |
| SfD2 | nd | sugar |
| SfD3 | 10.6 | sugar |
| SfD4 | 10.6 | sugar |
| SfD5 * | 0 | - |
| SfD6 * | 0 | - |
| SfD7 | 6.8 | sugar |
| SfD8 | 9 | sugar, concentrated orange juice |
| SfD9 | 11 | sugar, HFCS |
| SfD10 | 12.9 | HFCS, concentrated orange juice |
| SfD11 | 12.9 | HFCS |
| SfD12 | 10.64 | sugar |
| SfD13 * | 0 | - |
| SfD14 | 10.1 | sugar |
| SfD15 | 9.5 | concentrated grape juice, sugar, HFCS |
| Fruit drinks | ||
| Apple drink 2 | 9.9 | concentrated apple juice, sugar, HFCS |
| Apple drink 3 | nd | concentrated apple juice |
| Apple-mint drink 4 | 9.1 | concentrated apple juice, sugar, HFCS |
| Banana-lemon drink1 | 14 | banana pomace, HFCS, concentrated lemon juice |
| Cherry-apple drink 4 | 9.8 | concentrated apple juice, sugar, HFCS, concentrated aronia juice |
| Green Tea Lemon drink 5 | 4.6 | sugar |
| Lemon drink 5 | 6.9 | sugar, concentrated lemon juice |
| Peach drink 5 | 5 | sugar, concentrated peach juice |
| Green Tea drink 5 | 6.8 | sugar, HFCS |
| Lemon drink 6 | 7 | sugar, concentrated lemon juice |
| Peach drink 6 | 6.6 | sugar, concentrated peach juice |
| Orange drink 2 | 9.8 | concentrated orange juice, sugar, HFCS |
| Black tea & peach drink 7 | 7.2 | sugar, HFCS, concentrated peach juice |
| Green tea&strawberry | 7.7 | sugar, HFCS, concentrated strawberry juice |
| Fruit nectars | ||
| Banana 4 | 11.8 | banana pomace, sugar |
| Black currant 4 | 13.2 | concentrated black currant juice, sugar, HFCS |
| Black currant 5 | 12.3 | concentrated black currant juice, HFCS |
| Black currant 6 | 11 | concentrated black currant juice, sugar |
| Grapefruit 1 | 9.2 | HFCS |
| Orange 4 | 11.2 | concentrated orange juice, sugar |
| Red orange 4 | 11.4 | concentrated orange and carrot juice, sugar |
| Fruit juices | ||
| Apple antonovka 4 | 10.4 | juice reconstituted from apple concentrate, apple pomace |
| Apple antonovka 5 | 10.2 | juice reconstituted from apple concentrate, apple pomace |
| Apple 1 | 11.3 | juice reconstituted from apple concentrate |
| Apple 4 | 10.2 | juice reconstituted from apple concentrate |
| Apple 5 | 10.8 | juice reconstituted from apple concentrate |
| Apple 6 | 9 | juice reconstituted from apple concentrate |
| Apple 7 | 11.2 | natural apple juice |
| Birch juice | 5.43 | natural birch juice, sugar |
| Birch juice with aronia | 7 | natural birch juice, aronia juice, sugar |
| Orange 1 | 10.9 | juice reconstituted from orange concentrate |
| Orange 3 | nd | juice reconstituted from orange concentrate |
| Orange 4 | 10.2 | juice reconstituted from the orange concentrate |
| Orange 5 | 9.7 | juice reconstituted from orange concentrate |
| Orange 6 | 9 | juice reconstituted from orange concentrate |
| Orange 7 | 9.8 | natural orange juice |
| Orange 8 | 9.3 | natural orange juice |
| Orange 9 | 9.5 | juice reconstituted from orange concentrate |
| Pineapple 4 | 12.2 | juice reconstituted from pineapple concentrate |
| Pineapple 6 | 12 | juice reconstituted from pineapple concentrate |
| Red grapefruit 4 | 9.5 | juice reconstituted from red grapefruit concentrate |
| White grapefruit 5 | 9 | juice reconstituted from grapefruit concentrate |
| White grapefruit 6 | 9 | juice reconstituted from white grapefruit concentrate |
1–9—various producers. *—sugar-free product according to declaration. ED = energy drinks; SD = sports drinks; SfD = soft drinks. 1–17—particular products. nd—not determined by the manufacturer.
Table A2.
Concentrations of fructose, glucose, sucrose, and total sugars in the analyzed beverages (g/100 mL) (average ± SD, range and median calculated for the group of products in bold).
Table A2.
Concentrations of fructose, glucose, sucrose, and total sugars in the analyzed beverages (g/100 mL) (average ± SD, range and median calculated for the group of products in bold).
| Product | Fructose | Glucose | Sucrose | Total Sugars Determined Concentration |
|---|---|---|---|---|
| (g/100 mL) | (g/100 mL) | |||
| Energy drinks | 4.29 ± 1.71 | 6.33 ± 1.70 | 4.83 ± 2.08 | 14.2 ± 3.45 |
| 0.12–7.32 | 3.46–10.1 | 1.4–7.47 | 3.32–17.2 | |
| (4.86) | (6.44) | (5.39) | (15.6) | |
| ED1 | 5.67 ± 0.14 | 6.85 ± 0.19 | 2.82 ± 0.08 | 15.3 |
| ED2 | 5.22 ± 0.48 | 5.91 ± 0.54 | ND | 11.1 |
| ED3 | 3.92 ± 0.05 | 4.38 ± 0.06 | 6.56 ± 0.06 | 14.9 |
| ED3 * | ND | ND | ND | ND |
| ED4 | 5.32 ± 0.29 | 7.21 ± 0.29 | 3.08 ± 0.14 | 15.6 |
| ED5 | 5.26 ± 0.03 | 6.00 ± 0.03 | 4.42 ± 0.1 | 15.7 |
| ED6 | 5.20 ± 0.17 | 10.11 ± 0.6 | 1.40 ± 0.15 | 16.7 |
| ED7 | 4.67 ± 0.06 | 5.16 ± 0.10 | 6.98 ± 0.14 | 16.8 |
| ED8 | 7.32 ± 0.79 | 8.10 ± 0.68 | 1.76 ± 0.19 | 17.2 |
| ED9 | 1.92 ± 0.03 | 4.77 ± 0.04 | 7.47 ± 0.14 | 14.2 |
| ED10 | 3.35 ± 0.22 | 6.44 ± 0.37 | 5.99 ± 0.07 | 15.8 |
| ED11 | 3.64 ± 0.13 | 7.21 ± 0.74 | 5.39 ± 0.49 | 16.2 |
| ED12 | 2.64 ± 0.07 | ND | 7.43 ± 0.13 | 10.1 |
| ED13 | 5.13 ± 0.09 | 7.96 ± 0.11 | 3.36 ± 0.06 | 16.5 |
| ED14 | 4.86 ± 0.43 | 4.81 ± 0.24 | 6.12 ± 0.23 | 15.8 |
| ED14 * | ND | ND | ND | ND |
| ED15 | 0.12 ± 0.01 | ND | 3.20 ± 0.03 | 3.32 |
| ED16 | 5.74 ± 0.2 | 6.63 ± 0.16 | ND | 12.8 |
| ED17 | 2.98 ± 0.01 | 3.46 ± 0.28 | 6.47 ± 0.29 | 12.9 |
| Sports drinks | 1.02 ± 0.91 | 2.63 ± 1.31 | 3.93 ± 1.63 | 5.59 ± 2.18 |
| 0.28–2.37 | 0.50–4.01 | 2.50–5.73 | 2.88–9.54 | |
| (0.46) | (2.62) | (3.74) | (5.84) | |
| SD1 | ND | 3.35 ± 0.12 | 2.59 ± 0.12 | 5.94 |
| SD2 * | ND | ND | ND | ND |
| SD3 | 2.37 ± 0.12 | 1.45 ± 0.15 | 5.73 ± 0.12 | 9.54 |
| SD4 | 1.55 ± 0.07 | 2.62 ± 0.02 | 2.50 ± 0.12 | 6.67 |
| SD5 | ND | 4.00 ± 0.14 | ND | 4 |
| SD6 | 0.46 ± 0.04 | 0.50 ± 0.02 | 4.89 ± 0.11 | 5.84 |
| SD7 | 0.42 ± 0.01 | 2.47 ± 0.02 | ND | 2.88 |
| SD8 | 0.28 ± 0.00 | 4.01 ± 0.07 | ND | 4.28 |
| Soft drinks | 5.77 ± 1.13 | 6.04 ± 0.88 | 1.13 ± 1.11 | 12.7 ± 1.30 |
| 3.72–7.48 | 4.30–7.81 | 0.13–2.91 | 10.9–14.7 | |
| (5.83) | (6.13) | (0.44) | (12.45) | |
| SfD1 | 6.53 ± 0.23 | 5.42 ± 0.18 | ND | 11.9 |
| SfD2 | 3.72 ± 0.16 | 4.30 ± 0.14 | 2.91 ± 0.13 | 10.9 |
| SfD3 | 5.77 ± 0.01 | 6.58 ± 0.1 | 0.44 ± 0.02 | 12.8 |
| SfD4 | 6.06 ± 0.21 | 6.83 ± 0.4 | 0.26 ± 0.02 | 13.2 |
| SfD5 * | ND | ND | ND | ND |
| SfD6 * | ND | ND | ND | ND |
| SfD7 | 5.12 ± 0.19 | 5.60 ± 0.15 | 0.38 ± 0.04 | 11.1 |
| SfD8 | 4.67 ± 0.31 | 5.29 ± 0.32 | 2.14 ± 0.24 | 12.1 |
| SfD9 | 6.44 ± 0.09 | 7.81 ± 0.15 | 0.20 ± 0.00 | 14.4 |
| SfD10 | 7.40 ± 0.7 | 6.19 ± 0.61 | ND | 13.6 |
| SfD11 | 7.48 ± 0.17 | 6.40 ± 0.26 | ND | 13.9 |
| SfD12 | 5.68 ± 0.24 | 6.06 ± 0.74 | 0.13 ± 0.04 | 11.9 |
| SfD13 * | ND | ND | ND | ND |
| SfD14 | 5.89 ± 0.19 | 6.26 ± 0.27 | 2.54 ± 0.17 | 14.7 |
| SfD15 | 4.50 ± 0.25 | 5.68 ± 0.25 | 1.20 ± 0.15 | 11.4 |
| Fruit drinks | 2.74 ± 2.20 | 2.71 ± 1.81 | 4.29 ± 1.89 | 9.74 ± 2.77 |
| 0.16–7.52 | 0.25–6.48 | 1.51–7.38 | 4.98–15.5 | |
| (1.85) | (2.14) | (3.97) | (9.50) | |
| Apple drink 2 | 4.57 ± 0.29 | 4.60 ± 0.08 | 2.34 ± 0.03 | 11.5 |
| Apple drink 3 | 7.52 ± 0.41 | 3.65 ± 0.12 | 1.51 ± 0.09 | 12.7 |
| Apple-mint drink 4 | 4.42 ± 0.34 | 3.94 ± 0.06 | 2.69 ± 0.11 | 11.1 |
| Banana-lemon drink 1 | 6.92 ± 0.29 | 6.48 ± 0.21 | 2.14 ± 0.18 | 15.5 |
| Cherry-apple drink 4 | 4.69 ± 0.4 | 4.65 ± 0.4 | 3.94 ± 0.14 | 13.3 |
| Green Tea drink 5 | 1.43 ± 0.24 | 0.42 ± 0.08 | 6.72 ± 0.26 | 8.6 |
| Green Tea Lemon drink 5 | 0.95 ± 0.02 | 0.97 ± 0.03 | 4.05 ± 0.15 | 6 |
| Lemon drink 5 | 0.99 ± 0.15 | 2.06 ± 0.19 | 7.38 ± 0.22 | 10.4 |
| Peach drink 5 | 0.52 ± 0.04 | 0.49 ± 0.05 | 3.97 ± 0.14 | 5 |
| Lemon drink 6 | 0.16 ± 0.01 | 0.25 ± 0.01 | 6.45 ± 0.14 | 6.9 |
| Peach drink 6 | 1.63 ± 0.13 | 1.85 ± 0.12 | 4.85 ± 0.23 | 8.3 |
| Orange drink 2 | 3.61 ± 0.16 | 4.58 ± 0.15 | 3.23 ± 0.14 | 11.4 |
| Black tea & peach drink 7 | 2.55 ± 0.05 | 3.20 ± 0.16 | 2.20 ± 0.19 | 8 |
| Green tea & strawberry drink 7 | 1.33 ± 0.01 | 3.73 ± 0.28 | 3.36 ± 0.11 | 8.4 |
| Fruit nectars | 4.49 ± 2.36 | 4.46 ± 2.13 | 5.56 ± 2.55 | 13.7 ± 4.05 |
| 1.83–8.48 | 2.12–7.52 | 0.70–7.79 | 8.06–21.1 | |
| (4.38) | (4.26) | (6.17) | (13.7) | |
| Banana 4 | 1.83 ± 0.04 | 2.37 ± 0.07 | 7.26 ± 0.36 | 11.46 |
| Black currant 4 | 8.48 ± 0.44 | 6.76 ± 0.1 | 5.86 ± 0.22 | 21.1 |
| Black currant 5 | 6.17 ± 0.46 | 7.52 ± 0.19 | ND | 13.69 |
| Black currant 6 | 5.02 ± 0.16 | 5.17 ± 0.28 | 5.26 ± 0.22 | 15.45 |
| Grapefruit 1 | 4.38 ± 0.91 | 2.99 ± 0.34 | 0.70 ± 0.16 | 8.06 |
| Orange 4 | 1.96 ± 0.05 | 2.12 ± 0.02 | 7.79 ± 0.22 | 11.88 |
| Red orange 4 | 3.61 ± 0.26 | 4.26 ± 0.31 | 6.47 ± 0.20 | 14.34 |
| Fruit juices | 4.68 ± 1.97 | 3.63 ± 1.26 | 3.38 ± 1.53 | 11.7 ± 2.74 |
| 2.20–7.86 | 2.42–7.11 | 1.28–6.91 | 7.33–19.4 | |
| (4.03) | (3.24) | (3.23) | (11.3) | |
| Apple antonovka 4 | 7.65 ± 0.05 | 3.56 ± 0.04 | 1.82 ± 0.08 | 13.04 |
| Apple antonovka 5 | 6.57 ± 0.24 | 2.48 ± 0.14 | 1.28 ± 0.07 | 10.32 |
| Apple 1 | 6.74 ± 0.31 | 3.83 ± 0.07 | 1.47 ± 0.05 | 12.04 |
| Apple 4 | 6.53 ± 0.22 | 2.71 ± 0.28 | 2.68 ± 0.13 | 11.92 |
| Apple 5 | 7.09 ± 0.24 | 2.79 ± 0.18 | 2.26 ± 0.23 | 12.14 |
| Apple 6 | 7.86 ± 0.09 | 2.90 ± 0.04 | 1.75 ± 0.05 | 12.51 |
| Apple 7 | 6.57 ± 0.11 | 2.42 ± 0.16 | 2.21 ± 0.30 | 11.19 |
| Birch juice | 2.20 ± 0.04 | 2.51 ± 0.05 | 2.62 ± 0.09 | 7.33 |
| Birch juice with aronia | 4.30 ± 0.42 | 4.59 ± 0.13 | 1.62 ± 0.08 | 10.5 |
| Orange 1 | 2.24 ± 0.06 | 2.44 ± 0.22 | 3.87 ± 0.06 | 8.55 |
| Orange 3 | 2.80 ± 0.26 | 2.94 ± 0.27 | 4.20 ± 0.36 | 9.94 |
| Orange 4 | 3.04 ± 0.03 | 3.20 ± 0.11 | 3.73 ± 0.40 | 9.97 |
| Orange 5 | 2.42 ± 0.06 | 2.72 ± 0.22 | 4.03 ± 0.09 | 9.17 |
| Orange 6 | 2.74 ± 0.36 | 2.96 ± 0.28 | 4.42 ± 0.37 | 10.12 |
| Orange 7 | 3.10 ± 0.09 | 3.55 ± 0.11 | 4.09 ± 0.03 | 10.74 |
| Orange 8 | 3.27 ± 0.11 | 3.49 ± 0.12 | 5.09 ± 0.26 | 11.85 |
| Orange 9 | 3.04 ± 0.03 | 3.28 ± 0.05 | 5.10 ± 0.03 | 11.41 |
| Pineapple 4 | 4.05 ± 0.14 | 5.02 ± 0.28 | 5.98 ± 0.10 | 15.05 |
| Pineapple 6 | 6.15 ± 0.29 | 6.30 ± 0.3 | 6.91 ± 0.17 | 19.36 |
| Red grapefruit 4 | 6.72 ± 0.02 | 7.11 ± 0.26 | 3.86 ± 0.17 | 17.69 |
| White grapefruit 4 | 4.01 ± 0.06 | 4.68 ± 0.07 | 2.72 ± 0.25 | 11.4 |
| White grapefruit 5 | 3.91 ± 0.48 | 4.37 ± 0.25 | 2.56 ± 0.02 | 10.84 |
1−9—various producers (products characteristics given in Table A1). *—sugar-free product. ND—not detected.
References
- ADA Reports. Position of the American Dietetic Association: Use of nutritive and nonnutritive sweeteners. J. Am. Diet. Assoc. 2004, 104, 255–275. [Google Scholar] [CrossRef]
- EFSA Panel on Dietetic Products, Nutrition, and Allergies (NDA). Scientific Opinion on Dietary Reference Values for carbohydrates and dietary fibre. EFSA J. 2010, 8, 1462–1539. [Google Scholar]
- Otten, J.J.; PitziHellwig, J.; Meyers, L.D. Dietary Reference Intakes (DRI). The Essential Guide to Nutrient Requirements; The National Academies Press: Washington, DC, USA, 2010. Available online: https://nap.nationalacademies.org/read/11537/chapter/1#iii (accessed on 15 August 2024).
- Dietary Guidelines for Americans (DGA). 2005. Available online: https://health.gov/sites/default/files/2020-01/DGA2005.pdf (accessed on 14 August 2024).
- Welsh, J.A.; Cunningham, S.A. The role of added sugars in pediatric obesity. Pediatr. Clin. N. Am. 2011, 58, 1455–1466. [Google Scholar] [CrossRef]
- Kostecka, J.; Jackowska, I.; Chabros, I.; Kostecka-Jarecka, J.; Kawecka, P.; Kostecka, M. Beverage Consumption and Factors Influencing the Choice of Beverages among Polish Children Aged 11–13 Years in 2018–2023. Nutrients 2024, 16, 1625. [Google Scholar] [CrossRef]
- Brisbois, T.D.; Marsden, S.L.; Anderson, G.H.; Sievenpiper, J.L. Estimated intakes and sources of total and added sugars in the Canadian diet. Nutrients 2014, 6, 1899–1912. [Google Scholar] [CrossRef]
- Kunachowicz, H.; Nadolna, I.; Przygoda, B.; Iwanow, K. Tabele Składu i Wartości Odżywczej Żywności; Wydawnictwo Lekarskie PZWL: Warszawa, Poland, 2016. (In Polish) [Google Scholar]
- Menezes, E.W.; de Melo, A.T.; Lima, G.H.; Lajolo, F.M. Measurement of carbohydrate components and their impact on energy value of foods. J. Food Comp. Anal. 2004, 17, 331–338. [Google Scholar] [CrossRef]
- Menezes, E.W.; Giuntini, E.B.; Dan, M.C.T.; Lajolo, F.M. New information on carbohydrates in the Brazilian Food Composition Database. J. Food Comp. Anal. 2009, 22, 446–452. [Google Scholar] [CrossRef]
- Chávez-Servín, J.L.; Castellote, A.I.; López-Sabater, M.C. Analysis of mono- and disaccharides in milk-based formulae by high-performance liquid chromatography with refractive index detection. J. Chromatogr. A 2004, 1043, 211–215. [Google Scholar] [CrossRef]
- Eyéghé-Bickong, H.A.; Alexandersson, E.O.; Gouws, L.M.; Young, P.R.; Vivier, M.A. Optimisation of an HPLC method for the simultaneous quantification of the major sugars and organic acids in grapevine berries. J. Chromatogr. B 2012, 885–886, 43–49. [Google Scholar] [CrossRef]
- Horwitz, W. Official Methods of Analysis of AOAC International, 17th ed.; AOAC International: Gaithersburg, MD, USA, 2002. [Google Scholar]
- Jovanovic-Malinovska, R.; Kuzmanova, S.; Winkelhausen, E. Oligosaccharide profile in fruits and vegetables as sources of prebiotics and functional foods. Int. J. Food Prop. 2014, 17, 949–965. [Google Scholar] [CrossRef]
- Khajehei, F.; Merkt, N.; Claupein, W.; Graeff-Hoenninger, S. Yacon (Smallanthus sonchifolius Poepp. & Endl.) as a novel source of health promoting compounds: Antioxidant activity, phytochemicals and sugar content in flesh, peel, and whole tubers of seven cultivars. Molecules 2018, 23, 278–297. [Google Scholar] [CrossRef]
- Medeiros, P.M.; Simoneit, B.R.T. Analysis of sugars in environmental samples by gas chromatography–mass spectrometry. J. Chromatogr. A 2007, 1141, 271–278. [Google Scholar] [CrossRef] [PubMed]
- Molnár-Perl, I. Simultaneous quantitation of acids and sugars by chromatography: Gas or high-performance liquid chromatography? J. Chromatogr. A 1999, 845, 181–195. [Google Scholar] [CrossRef]
- Walker, R.W.; Goran, M.I. Laboratory determined sugar content and composition of commercial infant formulas, baby foods and common grocery items targeted to children. Nutrients 2015, 7, 5850–5867. [Google Scholar] [CrossRef]
- Walker, R.W.; Dumke, K.A.; Goran, M.I. Fructose content in popular beverages made with and without high-fructose corn syrup. Nutrition 2014, 30, 928–935. [Google Scholar] [CrossRef]
- Wang, X.; Chen, Y. Determination of carbohydrates as their p-sulfophenylhydrazones by capillary zone electrophoresis. Carbohydr. Res. 2001, 332, 191–196. [Google Scholar] [CrossRef] [PubMed]
- Dolan, J.W. Avoiding Refractive Index Detector Problems. LCGC N. Am. 2013, 30, 1032–1037. Available online: https://www.chromatographyonline.com/view/avoiding-refractive-index-detector-problems-0 (accessed on 10 June 2024).
- Bailey, B.; Plante, M.; Crafts, C.; Acworth, I. Advances in Universal Detection. Available online: http://www.dionex.com/en-us/webdocs/87448-PO-HPLC-Universal-Detection-30June2010-LPN2562-01.pdf (accessed on 10 June 2024).
- Eom, H.Y.; Park, S.Y.; Kim, M.K.; Suh, J.H.; Yeom, H.; Min, J.W.; Kim, U.; Lee, J.; Youm, J.R.; Han, S.B. Comparison between evaporative light scattering detection and charged aerosol detection for the analysis of saikosaponins. J. Chromatogr. A 2010, 1217, 4347–4354. [Google Scholar] [CrossRef] [PubMed]
- Jia, S.; Park, J.H.; Lee, J.; Kwon, S.W. Comparison of two aerosol-based detectors for the analysis of gabapentin in pharmaceutical formulations by hydrophilic interaction chromatography. Talanta 2011, 85, 2301–2306. [Google Scholar] [CrossRef]
- Márquez-Sillero, I.; Cárdenas, S.; Valcárcel, M. Comparison of two evaporative universal detectors for the determination of sugars in food samples by liquid chromatography. Microchem. J. 2013, 110, 629–635. [Google Scholar] [CrossRef]
- Konieczka, P.; Namiesnik, J. Quality Assurance and Quality Control in the Analytical Chemical Laboratory: A Practical Approach; CRC Press-Taylor & Francis Group: Boca Raton, FL, USA, 2009. [Google Scholar]
- Council Directive 2001/112/EC of 20 December 2001 Relating to Fruit Juices and Certain Similar Products Intended for Human Consumption. Available online: http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2002:010:0058:0066:EN:PDF (accessed on 16 August 2024).
- Food and Drug Administration. Nutrition Labeling of Foods. Available online: http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?CFRPart=101 (accessed on 3 June 2024).
- Neves, M.F.; Trombin, V.G.; Lopes, F.F.; Kalaki, R.; Milan, P. The Orange Juice Business. A Brazilian Perspective. Wageningen Academic Publishers, 2012. Available online: https://books.google.co.jp/books?id=1SHrExQdXfIC&printsec=frontcover&hl=zh-CN#v=onepage&q&f=false (accessed on 15 August 2024).
- European Commission. Regulation (EU) No 1169/2011 of the European Parliament and of the Council of 25 October 2011 on the Provision of Food Information to Consumers. Available online: https://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:32011R1169 (accessed on 10 June 2024).
- Jarosz, M.; Rychlik, E.; Stoś, K.; Charzewska, J. Normy Żywienia dla Populacji Polski i ich Zastosowanie; Narodowy Instytut Zdrowia Publicznego-Państwowy Zakład Higieny: Warszawa, Poland, 2020. (In Polish) [Google Scholar]
- Kowalska, A.; Sińska, B.; Kucharska, A.; Piechowiak, K.; Dąbrowa, K.; Paskal, W.; Szypowski, W.; Szypowska, A. Sugar sweetened beverages consumption among polish adolescents—A pilot study. Pediatr. Endocrinol. 2015, 14, 25–34. [Google Scholar] [CrossRef]
- Piekara, A.; Krzywonos, M. Assessment of the Frequency of Sweetened Beverages Consumption among Adults in Poland. Int. J. Environ. Res. Public Health 2021, 18, 7029. [Google Scholar] [CrossRef] [PubMed]
- White, J.S.; Hobbs, L.J.; Fernandez, S. Fructose content and composition of commercial HFCS-sweetened carbonated beverages. Int. J. Obes. 2015, 39, 176–182. [Google Scholar] [CrossRef] [PubMed]
- Grembecka, M.; Baran, P.; Błażewicz, A.; Fijałek, Z.; Szefer, P. Simultaneous determination of aspartame, acesulfame-K, saccharin, citric acid and sodium benzoate in various food products using HPLC–CAD–UV/DAD. Eur. Food Res. Technol. 2014, 238, 357–365. [Google Scholar] [CrossRef]
- Grembecka, M.; Szefer, P. Simultaneous determination of caffeine and aspartame in diet supplements and non-alcoholic beverages using liquid-chromatography coupled to Corona CAD and UV-DAD detectors. Food Anal. Method 2012, 5, 1010–1017. [Google Scholar] [CrossRef]
- Ventura, E.E.; Davis, J.N.; Goran, M.I. Sugar content of popular sweetened beverages based on objective laboratory analysis: Focus on fructose content. Obesity 2011, 19, 868–874. [Google Scholar] [CrossRef] [PubMed]
- White, J.S. High-fructose syrup—So, what is it exactly? In Dietary Sugars and Health; Goran, M.I., Tappy, L., Lê, K.-A., Eds.; CRC Press Taylor & Francis Group: Boca Raton, FL, USA, 2015; pp. 81–97. [Google Scholar]
- Sanz, M.L.; Villamiel, M.; Martinez-Castro, L. Inositols and carbohydrates in different fresh fruit juices. Food Chem. 2004, 87, 325–328. [Google Scholar] [CrossRef]
- Grembecka, M.; Lebiedzińska, A.; Szefer, P. Simultaneous separation and determination of erythritol, xylitol, sorbitol, mannitol, maltitol, fructose, glucose, sucrose and maltose in food products by high performance liquid chromatography coupled to charged aerosol detector. Microchem. J. 2014, 117, 77–82. [Google Scholar] [CrossRef]
- Türkmen, I.; Ekşi, A. Brix degree and sorbitol/xylitol level of authentic pomegranate (Punica granatum) juice. Food Chem. 2011, 127, 1404–1407. [Google Scholar] [CrossRef]
- Fakhouri, T.H.I.; Kit, B.K.; Ogden, C.L. Consumption of Diet Drinks in the United States, 2009–2010. NCHS Data Brief, No 109. 2012. Available online: http://www.cdc.gov/nchs/data/databriefs/db109.pdf (accessed on 14 August 2024).
- Zucconi, S.; Volpato, C.; Adinolfi, F.; Gandini, E.; Gentile, E.; Loi, A.; Fioriti, L. Gathering Consumption Data on Specific Consumer Groups of Energy Drinks. Supporting Publications 2013:EN-394. 2013. Available online: https://efsa.onlinelibrary.wiley.com/doi/epdf/10.2903/sp.efsa.2013.EN-394 (accessed on 14 August 2024).
- Statista. Available online: https://www.statista.com/statistics/1290812/poland-per-capita-soft-drink-consumption/ (accessed on 17 August 2024).
- Campos-Ramírez, C.; Camacho-Calderon, N.; Villagran-Herrera, M.E.; Aguilar-Galarza, A.; Anaya-Loyola, M.A.; Palacios-Delgado, J. Soft Drink Addiction Scale: Reliability and Validity Analysis in Young Mexican People. Beverages 2024, 10, 15. [Google Scholar] [CrossRef]
Figure 1.
HPLC-CAD chromatogram of a standard mixture of erythritol (ER), fructose (FR), glucose (GL), sucrose (SU), and inositol (IN) of 50 µg/mL. Separation was achieved on a Shodex Asahipak, NH2P-50 4E, column packed with 5 µm shell particles (4.6 × 250 mm), water/acetonitrile in a ratio of 25/75 v/v with isocratic run at a flow rate of 0.8 mL/min at 25 °C. An aliquot of 10 µL of sample solution was injected.
Figure 1.
HPLC-CAD chromatogram of a standard mixture of erythritol (ER), fructose (FR), glucose (GL), sucrose (SU), and inositol (IN) of 50 µg/mL. Separation was achieved on a Shodex Asahipak, NH2P-50 4E, column packed with 5 µm shell particles (4.6 × 250 mm), water/acetonitrile in a ratio of 25/75 v/v with isocratic run at a flow rate of 0.8 mL/min at 25 °C. An aliquot of 10 µL of sample solution was injected.
Figure 2.
HPLC-CAD chromatograms of selected energy drinks samples. Separation was achieved on a Shodex Asahipak, NH2P-50 4E, column packed with 5 µm shell particles (4.6 × 250 mm), water/acetonitrile in a ratio of 25/75 v/v with isocratic run at a flow rate of 0.8 mL/min at 25 °C. An aliquot of 10 µL of sample solution was injected. *—sugar-free product.
Figure 2.
HPLC-CAD chromatograms of selected energy drinks samples. Separation was achieved on a Shodex Asahipak, NH2P-50 4E, column packed with 5 µm shell particles (4.6 × 250 mm), water/acetonitrile in a ratio of 25/75 v/v with isocratic run at a flow rate of 0.8 mL/min at 25 °C. An aliquot of 10 µL of sample solution was injected. *—sugar-free product.
Figure 3.
Total sugar content in the analyzed beverages determined by HPLC-CAD compared to producer nutrition declaration (in %; Min—minimum average deviation from the declaration, Max—maximum average deviation from the declaration).
Figure 3.
Total sugar content in the analyzed beverages determined by HPLC-CAD compared to producer nutrition declaration (in %; Min—minimum average deviation from the declaration, Max—maximum average deviation from the declaration).
Table 1.
Validation data of the analytical methodology.
| Substance | Linearity | Fortification Levels (µg/mL) Orange Juice/Energy Drink | Recovery (%) | RSD Intra-Day (%) | RSD Inter-Day (%) | LOD (µg/mL) | LOQ (µg/mL) | ||
|---|---|---|---|---|---|---|---|---|---|
| Calibration Curve Range (µg/mL) | R2 | Calibration Curve | |||||||
| Erythritol | 1.0–150 | 0.9998 | y = 0.79x + 1.74 | 10/15 25/30 50/45 | 99.1/98.0 98.5/101 99.1/100 | 1.72 1.72 1.82 | 3.82 4.22 4.56 | 0.06 | 0.16 |
| Fructose | 1.0–150 | 0.9996 | y = 0.79x + 0.94 | 10/15 25/30 50/45 | 98.1/98.9 100/102 100/98.7 | 1.25 1.88 2.79 | 3.89 2.65 4.15 | 0.16 | 0.48 |
| Glucose | 1.0–150 | 0.9995 | y = 0.62x + 1.20 | 10/15 25/30 50/45 | 94.9/97.1 96.8/105 102/98 | 1.02 1.91 1.66 | 3.88 2.34 3.41 | 0.05 | 0.16 |
| Sucrose | 1.0–150 | 0.9999 | y = 0.79x + 0.97 | 10/30 25/60 50/90 | 96.7/101 98.6/98.4 98.5/96.0 | 2.49 2.08 2.25 | 3.4 3.02 2.86 | 0.07 | 0.21 |
| Inositol | 1.0–150 | 0.9998 | y = 0.92x + 0.58 | 10/15 25/30 50/45 | 101/99.7 103/100 103/99.9 | 1.84 1.95 2.46 | 2.76 2.78 3.02 | 0.09 | 0.26 |
Table 2.
Concentrations of fructose, glucose, sucrose, and total sugars in the analyzed groups of beverages (g/100 mL) (average ± SD, range, and median in brackets).
Table 2.
Concentrations of fructose, glucose, sucrose, and total sugars in the analyzed groups of beverages (g/100 mL) (average ± SD, range, and median in brackets).
| Product | Fructose | Glucose | Sucrose | Total Sugars Determined Concentration |
|---|---|---|---|---|
| (g/100 mL) | (g/100 mL) | |||
| Energy drinks | 4.29 ± 1.71 | 6.33 ± 1.70 | 4.83 ± 2.08 | 14.2 ± 3.45 |
| 0.12–7.32 | 3.46–10.1 | 1.4–7.47 | 3.32–17.2 | |
| (4.86) | (6.44) | (5.39) | (15.6) | |
| Sports drinks | 1.02 ± 0.91 | 2.63 ± 1.31 | 3.93 ± 1.63 | 5.59 ± 2.18 |
| 0.28–2.37 | 0.50–4.01 | 2.50–5.73 | 2.88–9.54 | |
| (0.46) | (2.62) | (3.74) | (5.84) | |
| Soft drinks | 5.77 ± 1.13 | 6.04 ± 0.88 | 1.13 ± 1.11 | 12.7 ± 1.30 |
| 3.72–7.48 | 4.30–7.81 | 0.13–2.91 | 10.9–14.7 | |
| (5.83) | (6.13) | (0.44) | (12.45) | |
| Fruit drinks | 2.74 ± 2.20 | 2.71 ± 1.81 | 4.29 ± 1.89 | 9.74 ± 2.77 |
| 0.16–7.52 | 0.25–6.48 | 1.51–7.38 | 4.98–15.5 | |
| (1.85) | (2.14) | (3.97) | (9.50) | |
| Fruit nectars | 4.49 ± 2.36 | 4.46 ± 2.13 | 5.56 ± 2.55 | 13.7 ± 4.05 |
| 1.83–8.48 | 2.12–7.52 | 0.70–7.79 | 8.06–21.1 | |
| (4.38) | (4.26) | (6.17) | (13.7) | |
| Fruit juices | 4.68 ± 1.97 | 3.63 ± 1.26 | 3.38 ± 1.53 | 11.7 ± 2.74 |
| 2.20–7.86 | 2.42–7.11 | 1.28–6.91 | 7.33–19.4 | |
| (4.03) | (3.24) | (3.23) | (11.3) | |
Table 3.
Realization of recommended daily energy requirements through consumption of one glass of the analyzed beverages (250 mL #) (mean ± SD, range).
Table 3.
Realization of recommended daily energy requirements through consumption of one glass of the analyzed beverages (250 mL #) (mean ± SD, range).
| Average Total Sugar Concentration per Glass (g/250 mL) | Energy per Glass a (kcal/250 mL) | % Total Energy Intake b (%) | |
|---|---|---|---|
| Energy drinks | 35.4 ± 8.63 8.30–43.00 | 142 ± 34.5 33.2–172 | 4.72 ± 1.15 1.11–5.73 |
| Sports drinks | 14.0 ± 5.45 7.2–23.9 | 55.9 ± 21.8 28.8–95.4 | 1.86 ± 0.73 0.96–3.18 |
| Soft drinks | 31.6 ± 3.24 27.3–26.8 | 127 ± 13.0 109–147 | 4.20 ± 0.43 3.63–4.90 |
| Fruit drinks | 24.4 ± 6.93 12.5–38.8 | 97.4 ± 27.7 50.0–155 | 3.25 ± 0.92 1.67–5.17 |
| Fruit nectars | 34.3 ± 10.1 20.2–52.8 | 137 ± 40.5 80.6–211 | 4.57 ± 1.35 2.69–7.03 |
| Fruit juices | 29.2 ± 6.86 18.3–48.4 | 117 ± 27.4 73.3–194 | 3.90 ± 0.91 2.44–6.45 |
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Grembecka, M.; Lebiedzińska, A.; Szefer, P. Application of HPLC Coupled with a Charged Aerosol Detector to the Evaluation of Fructose, Glucose, Sucrose, and Inositol Levels in Fruit Juices, Energy Drinks, Sports Drinks, and Soft Drinks. Beverages 2024, 10, 94. https://doi.org/10.3390/beverages10040094
AMA Style
Grembecka M, Lebiedzińska A, Szefer P. Application of HPLC Coupled with a Charged Aerosol Detector to the Evaluation of Fructose, Glucose, Sucrose, and Inositol Levels in Fruit Juices, Energy Drinks, Sports Drinks, and Soft Drinks. Beverages. 2024; 10(4):94. https://doi.org/10.3390/beverages10040094
Chicago/Turabian StyleGrembecka, Małgorzata, Anna Lebiedzińska, and Piotr Szefer. 2024. "Application of HPLC Coupled with a Charged Aerosol Detector to the Evaluation of Fructose, Glucose, Sucrose, and Inositol Levels in Fruit Juices, Energy Drinks, Sports Drinks, and Soft Drinks" Beverages 10, no. 4: 94. https://doi.org/10.3390/beverages10040094
APA StyleGrembecka, M., Lebiedzińska, A., & Szefer, P. (2024). Application of HPLC Coupled with a Charged Aerosol Detector to the Evaluation of Fructose, Glucose, Sucrose, and Inositol Levels in Fruit Juices, Energy Drinks, Sports Drinks, and Soft Drinks. Beverages, 10(4), 94. https://doi.org/10.3390/beverages10040094
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