Final End of Year Report
Throughout the 2018 – 2019 school year, I worked collaboratively with my faculty mentor, Dr. Elmer Mojica, on studying the chromatographic characterization of bee pollen from different countries around the world. The original overall objective was to determine the composition of different solvent extracts of bee products, specifically from the Philippines. This was successfully completed using the five solvents of methanol, ethanol, dichloromethane, hexane, and chloroform. This led to the natural curiosity to determine the antioxidant activity and phenolic content of said solvent extracts. Both objectives were met using the HP 6890 Gas Chromatography Mass Spectroscopy and a microplate reader where the DPPH and Folin-Ciolcalteu assays were performed. Being given the opportunity by the Undergraduate Research Initiative to brainstorm, conduct, and discuss my research via presentations and blog-posts has been a critical and valuable asset to my analytical research.
Before beginning the physical research in the lab, I established three major learning outcomes that I wanted to work diligently towards throughout the academic school year. The first goal I wanted to reach was the articulation of a proper research question with an effective objective. I wanted my research to have meaning and a purpose. For this reason, I chose to work with bee pollen in relation to the health and nutrition benefits that it could possibly have on humans. For example, working with the assay gave me an insight to the amount of antioxidant levels. Antioxidants prevent the formation of free radicals, which are often cancer-causing, in the human body. Establishing an understanding of what bee pollen consists of was imperative before moving onto further research. Next, I wanted to utilize an appropriate and efficient methodology to find an answer to my research question. I did so using instrumental methods and mass spectroscopy analysis. Lastly, I aimed to present the research in a conference setting. I applied with my research abstract and was gratefully accepted to present in an Undergraduate Poster Session at the American Chemistry Society Annual Conference in Orlando, Florida. I can successfully say that I accomplished all three of these outcomes.
An imperative part of my research was the collaboration between my faculty mentor and myself. Dr. Mojica first helped introduce me to the GC-MS instrument and the sample preparation methods involved such as extraction and filtration methods using solvents and bee pollen. Once I was able to acquire the compounds that make up the pollen, he challenged the research to go even further by teaching me how to determine the phenol content and antioxidant levels. Two assays were used with the Biotek Cytation instrument, DPPH and Folin-Ciocalteu. The DPPH assay involved dissolving the DPPH solution and pollen extracts in methanol on a microtiter plate. The Folin-Ciocalteu assay involved diluted methanol mixed with Na2CO3. The absorbance was then measured at 745 nm post-incubation. We gathered that the phenol content was highest when methanol was used for extraction and lowest when hexane was used. The antioxidant amount was highest when extracted with chloroform and lowest when extracted with methanol. Without Dr. Mojica’s guidance and assistance in the assay processes, the procedure would not have reached the scientific caliber that it did. By meeting with him on a regular basis to discuss all aspects of the research, I heightened my ability to work with others in terms of problem-solving and effective communication.
The main initiative for my research project was being able to find an answer to the question: what components make up bee pollen? I wanted the answer to this to be used in future studies for human health benefits. While my study did not focus on how bee pollen benefits humans, it focused rather on what bee pollen consists of. For this reason, further analysis can be used to interpret the benefits of these components, specifically in terms of antioxidants. Although problems were encountered during the school year, most were fixed through collaborative problem solving. For example, during a reading of bee pollen in hexane, no peaks were found. Through doing several trials, we realized that a greater amount of hexane, in microliters, must be injected into the GC-MS for a proper reading to be established. We therefore injected 10 microliters as opposed to 5 microliters. For precise readings, it was necessary that trials be taken multiple times. It was then found that the non-polar solvents contained mainly non-polar compounds, specifically long-chain hydrocarbons.
The Undergraduate Research Initiative provided me with the motivation to ask, learn, and challenge my previous bee exploration. By collaborating with Dr. Mojica, I established a solidified understanding of the GC-MS instrumentation, filtering process, and two different assay methods. Through constant communication, we were able to establish a proper methodology and ability to solve rising issues in the lab. The conducting of chromatographic research on bee pollen has served as an educational and eye-opening experience with results that can be implemented by future researchers.