Carbohydrate tests reveal acetylsalicylic acid alters sugars composition of Bellis perennis
Abstract
By: Liz Venettis, revised by Sarah Goins, re-revised by The Biles
We hypothesized that acetylsalicylic acid would prolong the health of daisies, but Floralife would keep the daisies healthy longer. We tested the effects of acetylsalicylic acid and Floralife on cut daisies, Bellis perennis (Hackney 2003). By adding these substances to the plants we found Floralife was best able to preserve flowers, while in fact aspirin caused premature wilting. We performed a Benedict’s test for reducing sugars, a Barfoed’s test for monosaccharide reducing sugars, a Selivanoff’s test for ketoses and aldoses, a Bial’s test for furanose rings, and an Iodine test for the presence of starch. We also performed a paper chromatography test that separates the pigments present in the daisies, an absorbance spectrum that tests which ranges of light are being absorbed, and a test for the presence of PPO. The results showed that reducing sugars and starches are not present in any treatments, and both the control and the Floralife solutions contain ketoses and furanose rings, while the aspirin solutions contain aldoses and pyranose rings. The paper chromatography test showed only one pigment, presumably the same pigment, was present in all of the solutions. We found that Floralife absorbed within the normal ranges for photosynthesis, 400-500nm and 650-700nm, for plants while the other two specimens did not. According to the enzyme test we found that polyphenoloxidase was not present in any of our three samples. Differences in the sugar composition due to Aspirin suggest that Floralife is a more useful way to prolong the health of a cut flower.
Discussion:
By Taisa Tomycz, revised by Adam Nack, re-revised by The Biles
The goal of our experiment was to determine the effects of Floralife and Aspirin on daisies. According to our hypothesis, we expected that the flowers with aspirin would have a longer life and be healthier than those with plain tap water, but a shorter, unhealthier life than those treated with Floralife. Assuming that the health of a plant would interfere with its life processes, we decided to test this hypothesis by performing a series of tests designed to determine the composition of the daisies that were treated with Floralife and Aspirin.
Jasmonic acid is a hormone secreted by a plant when it is attacked. “It works a little like a shot of pain,” says Dr. Ralph Backhaus of the University of Arizona (1998). Using Aspirin on plants inhibits this signal in the same way it does in animals. Aspirin binds to the enzyme that produces jasmonic acid, rendering it inactive (Backhaus, 1998).
However, our research indicated that many plants, in fact, produce a form of non-acetylated aspirin (salicylic acid) to help fight off infection (EHC, 1998). These plants use aspirin in the same way that those, studied by Dr. Backhaus, use jasmonic acid. We decided to test this with our hypothesis.
From visual examination, we noticed that the Floralife flowers appeared very healthy; they were green, moist, and strong. Aspirin appeared unhealthy; the flowers were wilted, brownish, and very dry. This indicated to us that aspirin was less effective than Floralife, which we had expected. What was unexpected, however, was that water appeared healthier than Aspirin. There was little wilting, the plants were green, and they were only slightly brittle. This indicated that the aspirin, in fact, had an adverse affect.
There was a higher water level in the Aspirin group. This could be explained by the fact that our Aspirin solution was saturated and may have been hypertonic to the daisies, thus pulling the water from the cells of the plant. Since we observed the stems of the Aspirin group becoming brown, it is possible that the cells were drying out and dying. This hypothesis is supported by research conducted by M. S. Neff on cut roses, which found that saturated sugar solutions appeared to result in excessive wilting (Neff 1942). So the saturated nature of the Aspirin solution could ultimately have been the cause of the extreme wilting and dryness of the daisies.
While three of our carbohydrate tests turned up negative for all subjects, there were some interesting results. From our results all we could infer that the daisies do not contain the sugars that were being tested.
Benedict’s test indicated that there were no reducing sugars present in the daisies in detectable amounts. Barfoed’s test confirmed this result by testing for reducing monosaccharides: none were present. The iodine test indicated that there was no starch to be found in the plants, which was unexpected, but because all three groups tested negative for the presence of starch, the results of the iodine test were inconclusive.
Selivanoff’s test indicated that there was a difference in the groups. Both Floralife and water tested positive for disaccharide ketoses, while Aspirin showed neither ketoses nor aldoses. This indicates that Aspirin possessed no sugars whatsoever, which was unexpected and may have been due to experimental error. However, if the findings were valid, it would indicate that Aspirin had used up its chemically stored energy almost completely, which would in turn indicate that it was unhealthier.
Bial’s test indicated that there were pentose furanose rings present in both Floralife and water. Again, Aspirin came up negative. This indicates that Aspirin possessed no furanose rings, which is to be expected if there is no sugar present.
The graph of the absorption spectrum of the three groups indicates that there was no difference in the specimen’s absorbance in any group. The reason there was no apparent change in this graph is probably due to the fact that all of the pigments remained in all of the specimens. This finding is supported by the results from our paper chromatography strips.
The paper chromatography strips from all groups showed only one similarly colored band. These tests were performed three times, only once did it yield any results. We feel that the reason the first two trials did not yield results was because we used the stems of the daisies to make our solution, while the third time, leaves were used. We felt that there was a possibility that the leaves of the daisies would contain more chlorophyll, which would produce darker solutions and react better with the phosphate buffer. We found none of the chlorophyll pigments that we expected. However, we did see a brown-yellow stripe that indicated a pigment known as Allagochrome. This pigment is normally blue-green, but when oxidized in solution it turns yellow-brown (Habermann 1973). The Rf values of these pigments were progressively lower from Floralife to Aspirin to control. This data indicates that the different solutes had varying effects on the solubility of this pigment.
The experiments to test for PPO turned up negative. This means there was no PPO present in any of our groups, and this data is inconclusive.
Our enzyme experiment dealt with adding PPO to our three solutions. The results of this experiment indicated that there was a general logarithmic trend in the reaction rates that reversed itself between Floralife and Aspirin. This data would need to be compounded with several other experiments to see if the trend holds. If the trend does in fact hold, then it would indicate that Aspirin affected PPO by forcing it to do a reaction opposite from the reaction that occurs in solution with Floralife.
A possible source of error in the carbohydrate section of the lab could be the ambiguous coloration of the Bial’s results in both the control and Floralife solutions. The coloration could have been interpreted as either olive green or olive brown, thus, making the results of the Bial’s test open to dispute. We determined by consensus that the color was in fact green and not brown.
For the chromatography portion of the lab, some possible sources of error could be due to an inadequate amount of leaves and stems needed to make the necessary concentration of solution. When these tests were performed, the leaves were very dry and limited in number, so extra phosphate buffer had to be added to make sufficient amounts of liquid. This additional phosphate buffer may have caused our solutions to be too diluted to reveal all of the pigments present.
A possible source of error in the enzyme portion of the lab could be the scalar values of the logarithmic trends, which vary greatly with each test and could make the results slightly misleading.
In conclusion, we found that the effect of aspirin on daisies is in fact negative from both health and ascetic perspectives. With the exception of absorbance of light, tap water seems preferable to aspirin. As expected, however, Floralife did prolong the health of the plant.
One possible way to expand on our results would be to perform the carbohydrates tests on solutions containing only Floralife plant food and Aspirin tablets, which would allow us to see whether these substances affected the results of our experiment.
A possible way to expand on our results would be to test different preservative brands against each other as opposed to against aspirin. It is possible that some brands may perform more optimally than others.
Our results demonstrate that saturated aspirin solution causes a plant to wilt prematurely. Another experiment that could be performed using this information would be to determine if different concentrations of aspirin would affect plant preservation in radically different ways. It may be found that lower concentrations of aspirin in fact do preserve plant health longer than ordinary tap water.
By: Liz Venettis, revised by Sarah Goins, re-revised by The Biles
We hypothesized that acetylsalicylic acid would prolong the health of daisies, but Floralife would keep the daisies healthy longer. We tested the effects of acetylsalicylic acid and Floralife on cut daisies, Bellis perennis (Hackney 2003). By adding these substances to the plants we found Floralife was best able to preserve flowers, while in fact aspirin caused premature wilting. We performed a Benedict’s test for reducing sugars, a Barfoed’s test for monosaccharide reducing sugars, a Selivanoff’s test for ketoses and aldoses, a Bial’s test for furanose rings, and an Iodine test for the presence of starch. We also performed a paper chromatography test that separates the pigments present in the daisies, an absorbance spectrum that tests which ranges of light are being absorbed, and a test for the presence of PPO. The results showed that reducing sugars and starches are not present in any treatments, and both the control and the Floralife solutions contain ketoses and furanose rings, while the aspirin solutions contain aldoses and pyranose rings. The paper chromatography test showed only one pigment, presumably the same pigment, was present in all of the solutions. We found that Floralife absorbed within the normal ranges for photosynthesis, 400-500nm and 650-700nm, for plants while the other two specimens did not. According to the enzyme test we found that polyphenoloxidase was not present in any of our three samples. Differences in the sugar composition due to Aspirin suggest that Floralife is a more useful way to prolong the health of a cut flower.
Discussion:
By Taisa Tomycz, revised by Adam Nack, re-revised by The Biles
The goal of our experiment was to determine the effects of Floralife and Aspirin on daisies. According to our hypothesis, we expected that the flowers with aspirin would have a longer life and be healthier than those with plain tap water, but a shorter, unhealthier life than those treated with Floralife. Assuming that the health of a plant would interfere with its life processes, we decided to test this hypothesis by performing a series of tests designed to determine the composition of the daisies that were treated with Floralife and Aspirin.
Jasmonic acid is a hormone secreted by a plant when it is attacked. “It works a little like a shot of pain,” says Dr. Ralph Backhaus of the University of Arizona (1998). Using Aspirin on plants inhibits this signal in the same way it does in animals. Aspirin binds to the enzyme that produces jasmonic acid, rendering it inactive (Backhaus, 1998).
However, our research indicated that many plants, in fact, produce a form of non-acetylated aspirin (salicylic acid) to help fight off infection (EHC, 1998). These plants use aspirin in the same way that those, studied by Dr. Backhaus, use jasmonic acid. We decided to test this with our hypothesis.
From visual examination, we noticed that the Floralife flowers appeared very healthy; they were green, moist, and strong. Aspirin appeared unhealthy; the flowers were wilted, brownish, and very dry. This indicated to us that aspirin was less effective than Floralife, which we had expected. What was unexpected, however, was that water appeared healthier than Aspirin. There was little wilting, the plants were green, and they were only slightly brittle. This indicated that the aspirin, in fact, had an adverse affect.
There was a higher water level in the Aspirin group. This could be explained by the fact that our Aspirin solution was saturated and may have been hypertonic to the daisies, thus pulling the water from the cells of the plant. Since we observed the stems of the Aspirin group becoming brown, it is possible that the cells were drying out and dying. This hypothesis is supported by research conducted by M. S. Neff on cut roses, which found that saturated sugar solutions appeared to result in excessive wilting (Neff 1942). So the saturated nature of the Aspirin solution could ultimately have been the cause of the extreme wilting and dryness of the daisies.
While three of our carbohydrate tests turned up negative for all subjects, there were some interesting results. From our results all we could infer that the daisies do not contain the sugars that were being tested.
Benedict’s test indicated that there were no reducing sugars present in the daisies in detectable amounts. Barfoed’s test confirmed this result by testing for reducing monosaccharides: none were present. The iodine test indicated that there was no starch to be found in the plants, which was unexpected, but because all three groups tested negative for the presence of starch, the results of the iodine test were inconclusive.
Selivanoff’s test indicated that there was a difference in the groups. Both Floralife and water tested positive for disaccharide ketoses, while Aspirin showed neither ketoses nor aldoses. This indicates that Aspirin possessed no sugars whatsoever, which was unexpected and may have been due to experimental error. However, if the findings were valid, it would indicate that Aspirin had used up its chemically stored energy almost completely, which would in turn indicate that it was unhealthier.
Bial’s test indicated that there were pentose furanose rings present in both Floralife and water. Again, Aspirin came up negative. This indicates that Aspirin possessed no furanose rings, which is to be expected if there is no sugar present.
The graph of the absorption spectrum of the three groups indicates that there was no difference in the specimen’s absorbance in any group. The reason there was no apparent change in this graph is probably due to the fact that all of the pigments remained in all of the specimens. This finding is supported by the results from our paper chromatography strips.
The paper chromatography strips from all groups showed only one similarly colored band. These tests were performed three times, only once did it yield any results. We feel that the reason the first two trials did not yield results was because we used the stems of the daisies to make our solution, while the third time, leaves were used. We felt that there was a possibility that the leaves of the daisies would contain more chlorophyll, which would produce darker solutions and react better with the phosphate buffer. We found none of the chlorophyll pigments that we expected. However, we did see a brown-yellow stripe that indicated a pigment known as Allagochrome. This pigment is normally blue-green, but when oxidized in solution it turns yellow-brown (Habermann 1973). The Rf values of these pigments were progressively lower from Floralife to Aspirin to control. This data indicates that the different solutes had varying effects on the solubility of this pigment.
The experiments to test for PPO turned up negative. This means there was no PPO present in any of our groups, and this data is inconclusive.
Our enzyme experiment dealt with adding PPO to our three solutions. The results of this experiment indicated that there was a general logarithmic trend in the reaction rates that reversed itself between Floralife and Aspirin. This data would need to be compounded with several other experiments to see if the trend holds. If the trend does in fact hold, then it would indicate that Aspirin affected PPO by forcing it to do a reaction opposite from the reaction that occurs in solution with Floralife.
A possible source of error in the carbohydrate section of the lab could be the ambiguous coloration of the Bial’s results in both the control and Floralife solutions. The coloration could have been interpreted as either olive green or olive brown, thus, making the results of the Bial’s test open to dispute. We determined by consensus that the color was in fact green and not brown.
For the chromatography portion of the lab, some possible sources of error could be due to an inadequate amount of leaves and stems needed to make the necessary concentration of solution. When these tests were performed, the leaves were very dry and limited in number, so extra phosphate buffer had to be added to make sufficient amounts of liquid. This additional phosphate buffer may have caused our solutions to be too diluted to reveal all of the pigments present.
A possible source of error in the enzyme portion of the lab could be the scalar values of the logarithmic trends, which vary greatly with each test and could make the results slightly misleading.
In conclusion, we found that the effect of aspirin on daisies is in fact negative from both health and ascetic perspectives. With the exception of absorbance of light, tap water seems preferable to aspirin. As expected, however, Floralife did prolong the health of the plant.
One possible way to expand on our results would be to perform the carbohydrates tests on solutions containing only Floralife plant food and Aspirin tablets, which would allow us to see whether these substances affected the results of our experiment.
A possible way to expand on our results would be to test different preservative brands against each other as opposed to against aspirin. It is possible that some brands may perform more optimally than others.
Our results demonstrate that saturated aspirin solution causes a plant to wilt prematurely. Another experiment that could be performed using this information would be to determine if different concentrations of aspirin would affect plant preservation in radically different ways. It may be found that lower concentrations of aspirin in fact do preserve plant health longer than ordinary tap water.
Figure 4: This picture is a comparison of our three specimens. It highlights the differences between the three specimens, in particular the difference between the daises treated with Floralife and those treated with Aspirin. Notice the extreme wilting and brown discoloration of the stems on the flowers on the right side of the picture, which were the ones treated with Aspirin. The flowers on the left, which were put in water, and the flowers in the middle, which were treated with Floralife demonstrate the extreme contrast between the two specimens just mentioned and the specimen treated with aspirin. The flowers on the left and in the middle still appear to be healthy, with very minimal wilting, and green stems.