Sugar, Photosynthesis and Protein Tests of Generic and Brand Name Feverfew Pills Maintain Cost Does Not Yield Value

By: Becky Barr, Andrea Kendziorski, Jennifer Martin, and Emily Schafer

Abstract

Two types of herbal aids on the current pharmaceutical market that claim specifically to reduce the pain of migraine headaches were analyzed. A generic feverfew supplement, GNC, and a brand name, Nature’s Way, were tested to determine if higher price reflected higher quality, leading to greater effectiveness. We predicted a higher cost would have no effect on pain relief. Five carbohydrate tests, Benedict’s, Barfoed’s, Selivanoff’s, Bial’s, and Iodine, were performed to see if both pills were sugar free, as boasted on both labels. All tests turned negative, excepting Selivanoff’s. All samples analyzed in Selivanoff’s Test changed color, from clear to red, after approximately one minute. This indicated the presence of disaccharide ketoses, and therefore sugar, in both pills; contrary to their claims, though supportive of our hypothesis. Pigment separation through paper chromatography was then carried out to determine pigment similarities in the pills. This test was inconclusive; no pigments separated out onto the chromatogram strips. The Bradford Assay was subsequently administered to determine pill protein concentration. Ingesting drugs containing protein inhibiting agents stop the inflammation which leads to migraines, arguably a method in reducing migraine pain. Therefore, finding additional protein in the pills would be detrimental in fighting migraine pain. Results were inconclusive, as some curve values were negative, and others were sporadic. Two volunteers also took the supplements, finding them ineffective in stopping their migraines. We have here no experimental evidence showing differences between pills. Matching our prediction, paying more does not offer a quicker end to migraine pain.

Figure 6: Selivanoff’s test results shown after the completion of the experiment. Selivanoff’s test, indicates the presence of an aldose or a ketose. If a sample turns red in less than one minute, then a monosaccharide ketose is present. If it turns red in approximately one minute, it is a disaccharide ketose. Finally, if the sample turns red in greater than one minutes time, it is an aldose. Figure 6A-C are the Nature’s Way trials, these all turned red in approximately one minute. Figure 6D-F, are the GNC trials, these also turned red in approximately one minute. Figure 6G, was fructose, the control. At over one minute of time, the control did change to a light peach color, indicating an aldose. Both the samples being tested contain the same type of sugar, a disaccharide ketose, disproving the label which states the supplements are sugar free.

Discussion

Between ten and fifteen percent of the population suffers from migraines, but many do not seek medical treatment (Diener, 2000). Herbal supplements can be used as a remedy for the painful effects of these debilitating types of headaches. A major component of the supplements is an extract of the plant Feverfew, which helps relieve “inflammation and hot swelling,” (McKenna, 2002). Feverfew extract has anti spasmodic and antiaggregatory properties that suggest it may be effective for relieving disorders with inflammatory and contractile responses, such as migraines (McKenna, 2002). Like many drugs on the market, there are name brand and generic brands containing Feverfew that are available for the customers to buy to help alleviate their migraine pain. Generally, the generic brand costs less money than the name brand pills. Many people wonder if in fact the name brand pills and the generic brand pills contain the same ingredients. We tested a name brand called Natures Way and a generic GNC brand to determine if they contained the same ingredients that were possible to test for the presence. We also tested the effectiveness of the pills in two volunteer subjects. Our group hypothesized that the organic pills would contain the same ingredients, thereby showing the lack of difference between generic and name brand and price. We also hypothesized that because of the properties of Feverfew, those who tested both pills would experience a moderate amount of relief from migraines. After testing the name brand and generic brand organic Feverfew pills, we discovered that they contain the same ingredients. Both the name brand and generic brand did not, however, work in reducing the symptoms of migraines in our study. We hypothesized that there would be low protein concentrations in our solutions, indicating the presence of anti-inflammatory properties of the Feverfew.

There were a number of tests performed to study the similarities in the pills. First, we performed five sugar tests on both the Natures Way and GNC brands using three trials of both drugs. The five sugar tests were: “Benedict’s Test” testing for reducing sugars, “Barfoed’s Test” for monosaccharides, “Selivanoff’s Test” for aldose or ketose sugars, “Bial’s Test” for furanose, and “Iodine Test” for starch (Krha, 2003). We predicted the five sugar tests to all result in no change, indicating an absence of sugar in the pills. This was hypothesized because both the GNC brand and the Natures Way brand claimed that the pills were sugar free on the bottle labels. Our group assumed that the companies marketing the drugs were truthful in their labeling and therefore based our hypothesis on the claim that the sugar tests would result in no changes. The results of four sugar tests showed no change in both pill brands. The test that did show a positive change for both pill types, however, was “Selivanoff’s Test.” The change in that test showed disaccharide ketoses were present in both types of pills. The lack of color change in the other four sugar tests showed that the pills did not contain reducing sugars, monosaccharides, or starches (Krha, 2003). A disaccharide ketose presence should not turn positive in “Benedict’s“, “Barfoed’s“, “Starch“, or “Bial’s” Tests. Our hypothesis that both GNC and Natures Way pills contained the same ingredients was supported by the sugar tests. The hypothesis on the absence of sugar in the pills was not supported by the tests, given that a disaccharide ketose was present in the pills. The companies that market the pills were also, therefore, incorrect in their labeling on the pill bottles.

The next test performed to demonstrate similarities in the contents of the name brand and generic pills involved photosynthesis of the organic substances in the pills. A pigment identification test done using paper chromatography enabled us to determine the pigments present in the pills (Krha, 2003). We predicted that chlorophyll a and b would be represented on the chromatography strip for both pill samples. Our chromatography tests yielded inconclusive results. The chromatograms were run and no color bands were produced on the paper strips. Our hypothesis was not supported or refuted because of the inconclusive results of the chromatogram tests.

The final test performed on the pills was the Bradford Assay to determine protein concentration in both pill brands. Our group hypothesized that the protein concentration curve that resulted from the Bradford Assay would be lower than the BSA curve, therefore demonstrating the lack of high protein concentration in the pills. Low protein concentrations would indicate the presence of anti-inflammatory properties of the Feverfew. Our tests were once again inconclusive for this experiment. It was unclear if the dye CBBG bound to any proteins in the Feverfew solutions because there were vastly different outcomes for the various absorbance readings. The resulting Bradford Assay curve for both pill brands and for each trial performed yielded numbers that were very scattered on a data plot. For the most part, however, the absorbance readings of the Feverfew solutions were less than those for the standard BSA absorbance readings. We can therefore conclude that our hypothesis was generally acceptable because, even with scattered numbers, the absorbance values for the Feverfew solutions were less than the data for the standard BSA curve.

The efficiency of the Feverfew supplements was also tested in a mini-experiment conducted on two volunteers. One subject claims to average about three headaches per week, and the second subject about four per week. Both subjects take pills prescribed by their respective doctors at the onset of a migraine. The volunteers decided they would experiment with the effectiveness of the Feverfew by taking both pills for migraine relief, instead of their usual prescriptions. The first two weeks of the lab, the subjects took neither of the pills, and recorded their headache occurrences. The third and fourth weeks of lab, the subjects took Natures Way brand pills once everyday as recommended on the label and recorded their number of headaches. The subjects took the GNC brand pills once a day for the last two weeks of lab, and also recorded their headache frequencies. By the end of the six-week trial period, neither volunteer found relief from the Feverfew supplements. The first subject claimed three headaches the first two weeks, three the third and fourth weeks, and four headaches the fifth and six weeks. The second subject claimed four headaches the first two weeks, five the third and fourth weeks, and four the last two weeks. Subject one still averaged her normal four headaches per week, and subject two averaged her normal three headaches per week. Both volunteers also claimed no change in the intensity of their migraines. It is not indicated on the GNC and Natures Way brand pill bottles the amount of time necessary for the Feverfew extract to begin to affect the migraine occurrence in subjects.

Reasonable objections may be made regarding the results of our study. There are many more, even hundreds more types of herbal headache and migraine relief pills. These other types of pills may contain higher concentrations of sugars or proteins than the two pill varieties studied by our group. Different herbal pills that are more expensive may contain more of the anti-inflammatory proteins in Feverfew than the GNC and Natures Way brand tablets. Our group, due to time and financial restraints, only tested two types of pills, and therefore our results can only make conclusions regarding those particular remedies. Even though the subjects we tested for pill effectiveness claimed no relief using the GNC and Natures Way brands, there are countless people that swear by herbal migraine remedies. The volunteers for our experiment had prior biased opinions of the effective properties of herbal benefits since both subjects normally take prescription drugs for their migraine relief. We also had no knowledge of the necessary amount of time for the Feverfew extract to become effective in reducing inflammation.

There are various reasons for the inconclusiveness of our pigment testing and the Bradford Assay results. There were several reasons for the uncertain results of the pigment identification tests. The stock solutions that were dropped onto the chromatograms were made with water and therefore took much longer to dry. If the solutions had been made with ethanol, the drops would have dried much faster. Our stock solutions may have also not even contained chloroplasts in the first place. The contents of the pill gel caplets were made of dried Feverfew leaves. The chloroplasts most likely died or lost their pigments in the drying process. The Bradford Assay also yielded results that were inconclusive. The scattered and odd absorbance readings of the Bradford and pill solutions may have been affected by the time sensitivity of the dye used to determine protein concentrations. Our herbal solutions may have had an odd chemical interaction, unbeknownst to our lab group, with the Bradford or CBBG solutions and changed time sensitivity and therefore absorbance measurements of the solutions.

In conclusion, our tests produced results that somewhat varied with the initial hypotheses. The sugar tests showed that both GNC and Natures Way pills did not contain any sugars except for a disaccharide ketose. The presence of this disaccharide ketose went contrary to our initial hypothesis that no sugars would be present in each type of pill. The photosynthetic tests yielded no conclusive results since the chromatograms did not run properly. Protein concentrations in the Feverfew pills were also uncertain yet showed a general curve to be less than that of the standard BSA concentration curve. The Feverfew pills that we used did not work as a migraine preventative for the two subjects studied for the pill effectiveness. Our predictions were correct that the pill contents were consistent with each other, demonstrating that for these two brands, price was not an issue.