An Analysis of Gum and the Role Structures of Carbohydrates, Pigments, and Proteins Play in Longer Lasting Flavor

Team Eppendorf

Abstract

In investigating different properties of gum, we believed that different sugars and their varying composition played roles in determining the longevity of flavor in gum. The purpose of this study was to determine how certain brands of gum retain their flavor longer and compare the flavor difference between different brands of gum. The brands of gum involved and tested in this lab were Bubble Yum, Orbit, Juicy Fruit, Trident, and Dentyne Ice. Several tests were run to determine the carbohydrates found in gum including Barfoed’s, Benedict’s, Bial’s, and Selivanoff’s tests. It was determined that free ketones were present with monosaccharides, disaccharides, and polysaccharides; ketoses and aldoses were also found to vary with the different brands of gum. This helped to support our hypothesis by showing that sugars played a role in the flavor longevity of gum.

No photosynthetic pigments were found in cinnamon and mint leaves using the paper chromatography test; therefore, an Rf factor could not be determined. The Bradford Assay concluded that the protein amount was found in every brand tested; this also confirmed with our hypothesis.

A single-blinded random flavor longevity study was performed with twenty human subjects and Dentyne Ice was found to have the longest lasting flavor while Juicy Fruit had the least lasting flavor. The study was performed to help gauge how our scientific tests compare to our results. Our hypothesis was supported and showed that sugars may control the longevity of the lasting flavor.

Figure 1. Benedict’s Test. Bubble Yum – The picture on the left (Picture A) is showing the three trials run for Benedict’s test for Bubble Yum using a saliva solution. Benedict’s reagent and Bubble Yum were added to the saliva solution before the test was run. All three trials resulted in a reddish/brown color. This indicates that free aldehydes or ketones were present in all of the three trials of Bubble Yum. The picture on the right (Picture B) is showing the three trials that were run using distilled water. Bubble Yum and Benedict’s reagent were mixed with the distilled water before the test was run. Again, all three trials resulted in a brown/reddish color. This shows that free aldehydes or ketones were present in Bubble Yum.

Discussion

Which gum should be crowned king for its flavor longevity? That was the purpose and investigation of our experiments. According to our study participants, Dentyne Ice deserves the honor as longest lasting gum for flavor. The real question isn’t which gum’s flavor lasts the longest though. The question we pursued in this lab was why gum retains its flavor. We believed at the beginning of the experiment that different sugars and their structures played the role of determining flavor longevity while photosynthetic pigments, enzymes, and proteins had little or no role in the process.

Throughout our experiments and tests we compared five different brands of gum: Bubble Yum, Juicy Fruit, Dentyne Ice, Trident, and Orbit. We expected to find several similarities between the different brands of gum but also differences which would lead to support for what determines flavor longevity in gum. In order to answer this question we conducted an array of tests in order to discover what affects the flavor longevity of the five different types of gum. These tests were designed to characterize the carbohydrate compositions, protein content, and photosynthetic qualities of each type of gum. Also, a single-blinded human taste study was conducted. This allowed us to compare the scientific data to human judgment for flavor longevity. The results and their meanings are discussed below.

Benedicts Test
Carbohydrates consist of simple single-monomer sugars (monosaccharides), two–monomer sugars (disaccharides), and other multi-unit sugars (polysaccharides) (Campbell et al, 2000). They contain several different compounds including sugars, starches, and celluloses (Krha et al, 2003). If the gum contains reducing sugars (free aldehyde or ketone groups), a red color would form as the copper is reduced. If no reducing sugars are present the trial prepared remains blue. Bubble Yum and Juicy Fruit turned a reddish color, (Figure 2, Figure 3, Table 1) indicating that these two brands of gum contain reducing sugars. While the saliva solution trails showed an orange-brown color, the distilled water trials resulted in a dark red color. While they both tested positive for reducing sugars, salivary amylase may have been a factor for the different color results. Orbit, Trident, and Dentyne Ice showed to have no reducing sugars as all of their trials for both the control and saliva solutions remained blue (Figure 1, Figure 4, Figure 5, Table 1). When compared to the human flavor longevity study (Figure 21), the results of Benedict’s show that Dentyne Ice, Trident, and Orbit, which contain no reducing sugars, were the three longest lasting gums in the study. This can help to confirm with our hypothesis that sugars do play a role in the flavor longevity in gum. It does this by showing reducing sugars shorten the flavoring of gum while gums that do not have reducing sugars does the opposite and expands the flavor in gum.

Barfoed’s Test
In lower pH levels and with shorter incubation times, monosaccharides react to form copper ions. A rust color will show that monosaccharides were present and no reaction indicates that disaccharides and polysaccharides were present in the trials (Krha et al, 2003). We expected to see a mixture of both monosaccharides and di/polysaccharides as not gums are made of exactly the same sugars. All five brands of gum (Bubble Yum, Trident, Juicy Fruit, Orbit, and Dentyne Ice) that were tested remained blue (Figure 6, Figure 7, Figure 8, Figure 9, Figure 10, Table 2); they were all disaccharides or polysaccharides. It is a little suspicious that all of the bands of gum remained blue for Barfoed’s test although this could simply be because disaccharides and/or polysaccharides are more beneficial in maintaining flavor longevity than monosaccharides; hence their overwhelming presence in all of the gums. Since we do not know for sure if this is the reason why, Barfoed’s test can neither support nor refute our hypothesis.

Selivanoff’s Test
Selivanoff’s test was used to see the difference between ketoses and aldoses. If the solution turned red in under a minute, the sugar is a monosaccharide ketose; if it turned red in approximately one minute, the sugar was a disaccharide ketose and if the solution took longer than a minute to turn red it was an aldose. Bubble Yum and Juicy Fruit (Figure 11, Figure 13, Table 3) turned red in less than a minute showing that ketoses were present. Orbit and Dentyne Ice (Figure 12, Table 3) resulted in a red color in approximately one minute indicating that disaccharide ketoses were present. Trident (Figure 14, Table 3) was the only brand of gum that took over one minute to produce a red color showing that aldoses were present in that particular brand of gum. When comparing the results of Selivanoff’s test to the human flavor longevity study (Figure 21) a conclusion can be made. Bubble Yum and Juicy Fruit were determined to contain ketoses and were the two lowest gums for flavor longevity in the human study. Dentyne Ice and Orbit, the two highest gums for flavor longevity in the human study, were found to contain disaccharide ketoses. Trident, also a higher ranked gum, was found to contain aldoeses. It was found that disaccharide ketoses may provide the longest lasting flavor in gum while aldoses provide longer flavor than ketoses. The results of Selivanoff’s test help to support our hypothesis.

Bial’s Test
Bial’s test was conducted to see if any furanoses (five-membered rings) were in the gums tested. The gums tested were Bubble Yum, Dentyne Ice, Juicy Fruit, Orbit, and Trident. If a green color results then the gum contains a furanose. An olive green color indicates that there is a pentose-furanose present while a brown color shows the presence of a hexose-furanose. If the trials remain yellow then a pyronose is contained in that gum (Krha et al, 2003). Bubble Yum and Juicy Fruit (Figure 15, Figure 17, Table 4) both displayed a brown color indicating that hexose-furanoses were present. Pyranoses were found to be present in Orbit and Trident (Figure 18, Figure 19, Table 4) as no color change was noted. Dentyne Ice displayed a green color after running Bial’s test signifying that it contains furanoses. These results helped to confirm our hypothesis.

Paper Chromatography and Rf Factor
Each pigment has a unique Rf value in a certain solvent and conditions. The Rf value can be altered by such things as solute, pH levels, and temperature (Krha et al, 2003). We substituted mint leaves and ground-up cinnamon in this portion of our experiments as it proved too difficult to dissolve the gum well enough to run this test. We expected to find some pigments in the mint leaves but were not expecting any pigments in the cinnamon. No pigments were found in any of gums that were tested using Paper Chromatography (Figure 20, Figure 21). We were a little surprised to find no pigments in the mint leaves, but after running multiple trials and ensuring proper procedures we were confident that the test was run correctly. It was determined that photosynthetic pigments do not play an important role in terms of flavor longevity. This helps to support our hypothesis as we thought that sugars played the major role in flavor longevity determination.

Absorption Spectrum
By using a spectrophotometer we were allowed to compare the light absorbency between different brands of gum. The gums that were tested were: Bubble Yum, Dentyne Ice, Juicy Fruit, Orbit, and Trident. As in the Paper Chromatography test, cinnamon and mint leaves were used. This was done because it was too difficult to dissolve the gums to a satisfactory amount to run the tests. We expected the cinnamon and mint leaves to have similar absorbencies. The results for absorption spectrum (Figure 22, Figure 23, Table 5) were slightly similar however they portrayed the same pattern. The general pattern that they both displayed was the higher the wavelength (in nm). This helped to confirm our hypothesis as it was determined not to play a role in the determination of flavor longevity.

Bradford Assay
Bradford’s Protein Assay was used to determine the amount of protein in each of our five brands of gum (Bubble Yum, Juicy Fruit, Dentyne Ice, Orbit, and Trident) by using protein concentration compared to absorbance. A bovine serum albumin (BSA) standard curve (Figure 24, Figure 25, Table 6) was prepared, allowing us to compare the absorbencies of our gum products to determine their protein concentrations. A tread-line was found for the BSA Standard Curve and an equation was drawn from that tread-line. The protein amounts were then calculated from the equation using the averages from the two trials (Table 7). We expected to find very little or no proteins in all of the gums that we tested. As expected, the protein amount found in all the gums was little to none (Table 8) The highest protein amount was found in Juicy Fruit at 5.16 mg/ml. Several of our gums when calculated were found to have negative amounts of protein; these were considered to contain no protein (0.0 mg/ml). Since the protein amounts in all the brands of gum were insignificant or none they helped to support our hypothesis that the sugars play the main role in determining flavor longevity in gum.

Conclusion
When comparing Tables 1, 3, and 4 you can see a pattern emerging. Bubble Yum and Juicy Fruit both display similar properties. Both contain hexose rings and are both ketoses. Orbit and Trident also display similar properties. Each of them contained pryanose rings and are aldahydes. Dentyne Ice is similar to Orbit and Trident but contains furanose rings. We believe that hexose rings and ketoses break down faster due to the fact that Bubble Yum and Juicy Fruit take fourth and fifth place in terms of their flavor longevity. Since Orbit and Trident took third and fourth place, one could deduce that pyranose rings and aldahydes might break down slower than hexose rings and ketoses. Dentyne Ice was the forerunner in the human flavor longevity survey and was the only gum to stand out due to the fact it contained a furanose ring.

Protein could also play a role in how long flavor lasted in gum. Table 8 shows all the protein amounts calculated and found in Bubble Yum, Juicy Fruit, Dentyne Ice, Trident, and Orbit. As noted before, all five brands of gum were found to have little to no protein amounts in them. It is not very likely however that protein does in fact play a role in terms of flavor longevity. The protein percentages are not significant and noteworthy enough to make any assumptions about how they affect flavor longevity.

From our information and data that was gather in this lab we believe that the reason some gums last longer than others in how the gum’s sugar molecules are arranged. It appears that a furanose ring is most stable and does not break down as fast as pryanose rings. Hexose was found to be the least stable and may break down the most rapidly. Protein content and photosynthetic properties were found not play a significant role in flavor longevity. Therefore our hypothesis can neither be fully upheld or fully refuted. We did seem to find evidence from the sugar tests that indicate they determine the flavor longevity in gum but we were not able to fully answer how and why. Why reason may be because studies have been conducted and it is now possible to attach the gum’s flavoring to the water-insoluble restrainer in the gum. This provides a controlled but efficient release of the gum’s flavoring during chewing (Song, 1996). This could have caused our carbohydrate tests to give inaccurate results as the gum’s flavor was being manipulated to release the flavoring over time and skewing our data. It would also not support our hypothesis but provides a reasoning of why gum’s flavor has longer longevity.

Even though precautions were taken and precise as possible steps were used for the preparation and execution of all of the tests used, there are several possible areas and sources for error and miscalculations. One error involves the pre-planning of our experiment. It has been found that -glucans can be co-dried with sweeteners or co-evaporated with syrups to product sweeteners that can help to prolong the gums flavor (Yatka, 1996). Since this was known, we could of devised better pathways around this obstacle to provide more accurate results for out five brands of gum that were tested. These errors and missteps cold have potentially embellished or understated our results leading to the incorrect support or weaken our hypothesis. Human error could have also played a large role in our experiments. Not cleaning test tubes and other materials properly, not following directions to minute detail or incorrect observations are just a few examples where errors could have occurred. Despite our research, it was found that some gums do contain a delayed release system. The sweeteners aspartame and acesulfame undergo a series of physical modifications using coating and drying. They are coated by encapsulation, partially coated by agglomeration, and entrapped by absorption. Then they are co-dried and particle sized to produce release modified aspartame-acesulfame (Muhammad et al, 1997). Also in preparing the gum for the Bradford Protein Assay, we dissolved the five different brands of gum in 3 Molar Hydrochloric Acid. This could very easily of denatured the structure of the gum while it was being dissolved and caused the results for the test to be askew. While these are not all of the possible errors that could have occurred in this lab, they are the most likely and steps were taken through the experiments to reduce the chance of error to the lowest amount possible.

References

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Krha, Maleszewski, Wilterding, Sayed, Luckie (2003) Testing Organic Compounds. Photosynthesis. Enzymes. LBS-145 Cell and Molecular Biology Fall 2003.

Muhammad J, Yatka RJ, McGrew GN, Record DW, Broderick KB, Song, JH (1997) Chewing gum containing encapsulated aspartame/acesulfame K. Trends in Food Science & Technology 8: 197.

Song JH and Brokeriek KB (1996) Chewing gum with prolonged flavour intensity. Trends in Food Science & Technology. 7: 143.

Yatka RJ and Meyers MA (1996) Chewing gum containing -glucans. Trends in Food Science & Technology. 7: 272-273.

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