Analysis of simple sugars in children’s fruit drinks

1. Introduction

Drinks are one of the most popular beverages worldwide (Rohman et al., 2022), with the busy lifestyle and the increased welling for ready to eat and drink items, the sugar drinks industry has expanded globally with consumers’ preference to satisfy thirst (Arboleda et al., 2021). The children’s age group was also targeted within the food and beverages business (Marx et al., 2022). However, dealing with children’s beverages is so tricky in terms of manufacturing and marketing as it requires characters to grip the attention of children such as colour and taste (Naderer, 2021). Moreover, it also needs to be appealed to and approved by the parents (Hall et al., 2022). However, the consumption of soft drinks has been argued in many studies due to the impact of sugar on health. Sugar has been known as an energy provider with no nutritional value (Riediger wt al., 2021). Indeed, it can cause serious health problems such as being overweight putting the child at a higher risk to develop cardiovascular diseases, diabetes due to irregular insulin response, and dental problems such as dental caries and cavities (Potempa-Jeziorowska et al., 2022). The additional problems that the sugar intake cause during childhood are digestive system problems such as stomachaches, acidic digestive tract, and diarrhoea (Uddin and Rumman, 2022). On the same hand, advertising drinks as being suitable for children or having no added sugar can mislead parents to think that they are making healthier choices for their children (Richter et al., 2022). Moreover, it has been reported that the front package label and the juice and fruit image used confuse the parent who are seeking natural and low or no added sugar drinks (Duffy et al., 2021). Measuring sugar in food is an important step in beverage manufacturing to provide accurate data and keep the product under control and within legislation (Jurica et al., 2021). Still, many studies are arguing the reasons behind high sugar intake among this age group along and the need for clear labels to assist parents in making healthier choices (Roberton et al., 2021).

Aim of the research:? to investigate sugar intake in children and adolescents, analyse the free sugar content of fruit drinks advertised as being suitable for children, or having low / no added sugar, and study the performance of the analytical method in comparison to other methods.

2. Methodology and Experimental Design

2.1 Equipment and materials?????????

Table 3: Equipment and material used for the sugar determination experiment.

2.2 Protocol

Table 4: Contents of the Megazyme K-SUFRG Enzyme Assay Kit

22ml of distilled water was added to Bottle 2 and mixed to dissolve, then 20ml of distilled was added to bottle 6- freeze in 1ml aliquots. All bottles were vortexed to be mixed before use. The sample was diluted with distilled water to give a concentration of sugars between 0.05- 0.8g/ltr. A note of the dilution factor was taken. The turbid solutions were filtered before use.? After the solutions were prepared as directed in the Magazyme manual kit and aliquoted into Eppendorf tubes. 200μL of bottle 6 was added to the blank sucrose samples and the actual sucrose samples, a P200 variable pipette was used to take l00μL of the same samples and were put into respective cuvettes for both sucrose and the glucose/fructose analyses, mixed via aspiration, and incubated at room temperature for 5 minutes. Then? lO0μL of both reagents 1 and 2 were pipetted to all cuvettes, ?both blanks and test samples. Finally, deionized water was added the amount of deionized water used in this step was different between blanks and samples with a total volume of 2.4mL for all cuvettes. Next, the samples were in each cuvette by aspiration, to ensure all reagents and samples had been equally mixed together. After 3 min incubation, the cuvettes were taken to a spectrophotometer at 340m. Then, 20μL of reagent 3 was added to each cuvette to trigger the conversion of fructose to glucose-6-phosphate, mixed and incubated at room temperature for 5 minutes. Then A2 readings were taken using a spectrophotometer. The same method was applied for A3 readings.

2.3 Calculations?

The first initial reading of the spectrophotometer was the A1 reading, this reading occurred before any actual conversion of sugars to NADPH has occurred, so any detected absorbance is due to the background of reagents and cuvettes. So, there was a need to subtract the initial absorbance from the final samples to ensure the defection of the true conversion of sugars to NADPH.

In addition,? the following calculations were used If the sample had been diluted the result must be multiplied up by the dilution factor. ???????????

1. (A2-A1) sucrose sample –(A2-A1)blank sucrose =ΔAtotal glu

2. (A2-A1)glu/fru sample-(A2-A1)blank Glu/fru =ΔAglu

3. ΔA totalglu-ΔAglu =ΔAsucrose

4. (A3-A2)glu/fru sample-(A3-A2)blank Glu/fru =ΔAfru

5. g Sucrose/ltr= (1.315 xΔAsucrose

6. g D-glucose/ltr =0.6920 xΔAglu

7. g D- fructose/ltr =0.6978 x ΔAfru

3. Results

Figure 5: calculated and advertised sugar content in different drinks samples

The figure represents the mean of duplicated samples tested for free sugar content of fruit drinks advertised as being suitable for children after proper dilutions for the blackcurrant and orange high juice vs the advertised sugar content. The results showed that the difference between the calculated and the advertised sugar content is 5 g/100 mL on average, except for Sainsbury’s Orange High Juice and Tesco Orange High Juice No Added Sugar using a spectrophotometer.

?

4. Discussion

The results showed the calculated sugar content of the fruit drink samples was less than 8 g/100mL for total sugars and less than 1 g/100ml for those advertised as having no added sugars. . The European commission guideline tolerance mentioned that if the amount of sugar is less than 10g/100ml? then a ± 2 g ?difference is accepted (Gebhart, 2013). In compression with the advertised sugar content which was almost similar except for Sainsbury’s Orange High Juice with higher calculated reading by 10.15 g/100mL as absolute value difference and Tesco Orange High Juice No Added Sugar with 4.33 g/100mL using a spectrophotometerStudies have shown that drinks and other beverages marketed toward children were also using the concept of lower sugar or no added sugar content as it grips parents’ attention (Swift et al., 2018). Many brands were lurching their drinks with only 1 gram of sugar such as Rethink Brands as well as drinkable yoghurt and drinks (Moore et al.,2020).? However, Magriplis et al (2021) claimed that recent data showed a sugar intake range between 7% to 34.8% in children and from?15.4% to 29.6%?in adolescents?globally. On the other hand, replacing sugars with sweeteners in drinks is another misleading point for parents who focus on the No Added sugar labels reference. In addition, fruit images were found on drinks that contain no fruit and the word “juice” was found on fruit-flavoured drinks (Harris and Pomeranz, 2021). Therefore, it is arguable that drinks advertised as no added sugars are not the healthier choice for children. Therefore, it is very important to check the label’s sugar values (Musicus et al., 2022). Regarding the performance of enzyme assay kits and spectrophotometer, the variation between calculated and advertised sugar content was less than 1g/100ml except for two brands which showed large variation. Méndez et al (2021) contented that K-SUFRG Enzyme Assay Kit is an appropriate test for D-fructose, D-glucose, and sucrose in food samples. However, Enzyme Assay Kit cannot detect non-sugar sweeteners (Nguyen et al., 2021). Moreover, Mohammed et al (2021) stated that UV spectrophotometers can be used in the food industry to detect dietary sugars. However,? G?owacka et al (2021) highlighted that there is a must to consider that UV spectrophotometers use cuvettes with two sides that were designed to be the 'light path', where the light is going to be shone through and these were often marked with an arrow at the top of the cuvette (Michalec and Tymecki, 2018). The lab technicians need to line this up with the light source inside the spectrophotometer and the initially detected absorbance would be due to the background of reagents and cuvettes (Jesenkovi?-Habul et al., 2019). Given this, it is arguable that the misreading variation in some sample results could be due to human error in addition to the environmental effects on the photometer, and this could affect the accuracy of the data (reference). to improve this, Infrared Spectroscopy could be used (Ba?kan et al., 2016). Aouadi et al (2020) suggested that? Infrared Spectroscopy is the number of different sugar types in different drink samples with distinguished efficiency and accuracy. Moreover, PLS analysis can be used as a supplemental method for quantitative analysis. Interestingly,? Riswahyuli et al (2022) argued that combining the Attenuated total reflection (ATR) technique with Infrared Spectroscopy will enable sugar determination directly without sample preparation. Therefore, this would be so helpful in obtaining results using no reagents and thus fewer or no human errors. SACN guidelines for childhood sugar intake (2015) contended that daily sugar consumption for? Children from 7 to 10 should be no more than 24g and a?19g maximum for children from 4 to 6 (Keith et al., 2020). And the average total sugar on drinks labels was 5.7 g and the average sugar on drinks advertised as No Added Sugar was 0.8. therefore, having one drink, not being advertised as No Added Sugar a day should not necessarily be labelled as bad or unhealthy for children (Fleming‐Milici et al., 2022). Indeed, many studies nowadays are arguing the validity of market choices caused by no added sugar fruit drinks advertised as being suitable for this age group as they could have other sweetens or maintain a habit of having drinks that do not add any nutritional value. On another hand, Li et al (2022) claimed that sweet drinks should not be aimed at children as they can enhance picky eating, digestion problems tooth decay and obesity (Arora et al., 2022). Therefore, Pomeranz et al (2020) suggested that fresh fruit and vegetable are a better option, as they contain fibres, maintain healthier eating habits, and teach children to chew different textures and try different tastes (Gandhi, 2022). However, Harris and Pomeranz (2021) suggested that if there was a need to insert juice into the child’s diet, then it should be limited to fresh homemade 100% fruit juice especially citric fruit as they are rich with vitamins and has a low amount of sugars (Lu et al., 2021).

5. Conclusion

The calculated sugar content of the fruit drink samples agrees with the value on the label except for two samples. The findings for sugar drinks match other similar beverages marketed toward children. Regarding the UV Spectrophotometer method used, the possibility of human error can explain the variation of some readings. This suggests that practitioners must be aware that variation in results will require reperforming the experiments or using more advanced analytical methods that don’t require adding reagents or pretreating the samples such as NIR spectroscopy. SACN guidelines for childhood sugar intake (2015) suggest less than 19 g (5 cubes) are allowed for children from the 4-6 age group daily. Therefore, in practice, it is a must to consider that children are having a lot of other sources of sugar intake during the day. Thus, it is arguable that a cup of drink with 6 g of sugar can be considered a problem. Therefore, there is a need for alternatives, this could be changing the formula of the drinks to citrus fruit with no sugar and no additives or having fresh vegetables or fruit which will have positive health contributions to children.

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