The Chemical Difference Between Pancrease and Lipram

Becky Haugen
John Lambrix
Kimberly Fagerman
Tyler Alden




Abstract
By: John Lambrix – revised, Becky Haugen
Our goal in this lab is to compare Pancrease and Lipram to determine if they are chemically different.  Our hypothesis is that there will be a difference between the two.   First, we performed carbohydrate comparison tests on both drugs.  The tests we used were Benedict’s, for free aldehyde and ketone groups, Barfoed’s, to distinguish monosaccharides from disaccharides and polysaccharides, Selivanoff’s, to differentiate between aldoses and ketoses, and Iodine to test for the presence of starch.  We found that Pancrease contained a Ketose and Lipram did not.  We took the products of these tests and checked their absorptions on the spectrometer to determine concentrations.
Next we compared the absorption spectrums of Pancrease and Lipram after performing the Hill reaction on eight spinach leaves.  We covered spinach leaves with each drug and performed Hill reaction.  We found that the enzymes inhibited all leave’s photosynthetic abilities.  Using enzyme testing we determined in what pH Pancrease and Lipram worked best (Table 6).  We put each drug with starch and then performed Barfoed’s reaction on the product to test for the presence of monosaccharides.  We also performed Iodines test to see how much starch remained.  In the carbohydrate tests we found that there was a free Ketose group on Pancrease.  This was shown because Pancrease turned orange/red in the first minute of being in boiling water with Selivanoff’s solution.  From our results we were able to support our hypothesis that these two drugs were, in fact, not identical.

                FIGURE    1

Table 1:  Testing for Organic Compounds:  The data from the Benedict’s test shows that Copper is not reduced.  The Barfoed’s test results show that monosaccharides are not present.  The data from Selivanoff’s test show that Pancrease is a ketose. Lastly, the Iodine test shows that there were no reactions.  

Trial 1:                   
     Benedict's    Barfoed's    Selivanoff's    Iodine
Pancrease    No reaction    No reaction    Red-brown/Ketose    No reaction
Lipram    No reaction    No reaction    No reaction    No reaction
Trial 2:                   
     Benedict's    Barfoed's    Selivanoff's    Iodine
Pancrease    No reaction    No reaction    Red-brown/Ketose    No reaction
Lipram    No reaction    No reaction    No reaction    No reaction


Discussion
By:  John, Tyler, Becky, & Kim
Our hypothesis is that there is a difference between Lipram, a generic drug, and Pancrease, a name brand drug.  The FDA has researched this same topic through their own private studies.  Our goal is to test these drugs ourselves to compare the results and search for inconsistencies.  We believe there is a chemical difference between these drugs.  We compared the structure of the two drugs by checking for simple and complex carbohydrates.  We tested both drugs on spinach chloroplasts to determine if it affected photosynthesis.  Lastly, we compared the activity of the enzymes in the drugs and how environmental changes affect them.  To test the structure of the two drugs we decided to use Benedict’s, Barfoed’s, Iodine, and Selivanoff’s test.  We predicted that the results of the tests alone would be similar; however we expected the absorbencies to be different.  
Our results from the Benedict’s test, Barfoed’s, and Iodine, were identical.  In Benedict’s test, no reaction occurred, showing that both drugs had reducing sugars or contained a free aldehyde or ketone.  Barfoed’s test showed that both drugs contained no monosaccharides.  The results were negative.  When we introduced iodine into the stock solution of each drug, starch was not present.  Although there three tests had the same negative results, the absorbencies of these results showed variation, which helps support our hypothesis.
An even more exciting breakthrough in our research was when we did Selivanoff’s test.  Pancrease produced a red-brown precipitate which is evidence that it may have a ketose, whereas Lipram did not change color.  Therefore, Lipram did not contain a ketose.  These results also helped support our hypothesis.
The spectrometer readings that we took of the results from Benedict’s, Barfoed’s, Selivanoff’s and Iodine tests all indicated that there were differences in the drugs.  For Benedict’s and Barfoed’s test, Lipram had a higher absorbance, whereas for the other two tests, Pancrease had the higher absorbance levels.  A higher absorbency indicates that the solution is more concentrated.  These absorbency results basically contradict each other.  Although this does support our hypothesis, is shows inconsistency.
    To test the effects of Lipram and Pancrease on the proteins, lipids, and carbohydrates of chloroplasts, we soaked spinach leaves in our stock solutions for twelve hours.  This amount of time allowed the enzymes in the drugs to fully work on the components of the chloroplasts.  We also used the Hill Reaction to help show how it affected the photosynthesis of the leaves.  Our predictions were that Pancrease would disrupt the photosynthesis activity more than Lipram because it is the national brand drug.  The results of the Hill Reaction were that Lipram had lower absorbencies at red light, blue light, white light, and no light.  A lower absorbency implies that more photosynthesis is occurring in the tested chloroplasts.  Since Lipram had lower absorbencies this means that the drug did not breakdown the carbohydrates, lipids, and proteins of the chloroplasts as well as Pancrease. Also, we visually compared the leaves after they had been acted on by the drugs for twelve hours.  The leaves submerged in the Pancrease stock solution appeared spotted and darker green.  In the solution, the spinach leaves appeared light green and seemed to have little change.  These tests helped negate our null hypothesis, which states that the two drugs were identical.
    To test how environmental changes affected the enzyme activity of the drugs, we performed several pH and temperature tests.  Initially, to determine how effective the enzymes were on starch, we mixed starch with the separate drugs and performed Barfoed’s and Benedict’s tests on the resulting solutions.  Both tests were negative.  This illustrates that neither drug broke starch down to a monosaccharide.  These results are acceptable; however we believe that if we would have allowed the solutions to react longer, the drugs may have broken down the starch further.  Next, we tested the enzyme activity at different pHs.  We predicted the optimal pH for the drugs would be approximately two, because it is the pH of the stomach.  After testing the enzyme activity with starch at pH’s ranging from two to seven and a half, we concluded that the optimal pH for both drugs was at seven and a half.  These results are logical because seven and a half is closest to the pH of the duodenum.  Lastly, we tested for the optimal temperature of the enzymes.  Our predictions were that out of the hot bath (90 degrees Celsius), ice bath (0 degrees Celsius), and body temperature, the enzymes would work best at body temperature.  Our results were skewed because Pancrease worked best at body temperature but Lipram worked best at the ice bath temperature.  This is another example of the difference between the drugs.
    With this data, we found that we were able to refute our null hypothesis that the two drugs were chemically identical.  Our data supports our original hypothesis that the two drugs were different.  According to the laws of biology and physiology, we were able to use tests that resembled the conditions in the body and see how well the drug worked.  We used pH’s that resembled the stomach (acidic) and the blood (more neutral).  We used a water bath of 37 degrees Celsius to replicate body temperature.  The higher temp water bath did not work as well, because the enzyme was probably denatured.
Within any experiment, research teams will encounter sources of error.  Within our complex experiment, there were many possible sources of error.  The fact that we did not repeat our tests as many times as we would have liked to due to time constraints and budget, could have made our results less accurate.  Also, we may not have let the enzymes in our drugs react long enough in the enzyme lab to breakdown starch to its maximum capacity.  If we would have had the capabilities to test the packaging of the drugs, it would have made our results more factual.
If we were to replicate our experiment there would be a couple changes we would make.  We would have tested our drugs on the other organic molecules (lipids, proteins, etc.) other than simply starch.  Another word to the wise, repetition, repetition, repetition.  Lastly, if we had better resources such as money, equipment, and time, we would have tested our drugs more thoroughly rather than with just the three assigned labs.  For example, we would make a computer generated model of the two drugs to compare them structurally.