Final Blog Post: An Exotic Butter Formulation to Enhance Bacterial Resistance and UV Protection

Rudra Persaud

Over the course of this past 2018-2019 academic year, I have worked with Dr. Jamielee Rizzo in studying how to generate antimicrobial surfaces that exhibit UV resistance. The overall end goal of this project is to find a more natural, yet effective way to address wound healing. As I prepare to graduate in May, having this chance to work with Dr. Rizzo on a scientific research project has encouraged my interest in relevant work and given me the drive to pursue a Ph.D. in science.

Through all of the hard work put into this project and time spent doing background research, I find that there were many instances where I had to find a balance with research along with school work and daily life- but I can say that having this opportunity taught me how to adequately balance my time. One of the reasons I enjoyed working with Dr. Rizzo is because she encourages me to be diligent in my work and gave me the chance to answer thought-provoking questions we came up with together, I felt as though she was able to provide an authentic in-field research experience because often times the questions that are asked may not have an answer yet. Dr. Rizzo was able to give me a chance to think like a real scientist while providing a safe distance to guide me whenever I had my doubts. Another reason why I enjoyed working with Dr. Rizzo is that she encouraged me to participate in as many research presentations as possible. Just recently I had the opportunity to go to the national American Chemical Society symposium in Orlando to present my findings to thousands of scientist from all over the world.

Based on the work completed over the course of this academic year I was able to discover may-interesting facts from the protocol we implement with our butter and oil antimicrobial samples. Base butter by themselves, tamanu, ucuuba, and Aloe Vera, have a known effect when it comes to protecting against UV light and having antimicrobial resistance (see RP1, RP6, and RP11 respectively). In general, it is observed that samples that have a darker tint tend to have a better UV resistance, this is expected because in darker pigments absorb a wide spectrum of light. In fact, the most favorable butter base to use is uccuba butter and tamanu butter, as it has a dark brown tint when liquefied. Butters that naturally have a lighter tint like Aloe Vera tend to have relatively good antimicrobial resistance, having a range of clearance of about 1 cm naturally, but due to its tint does not have good resistance to UV light. As we add essentials oils to our butter bases, we can observe an increase in antimicrobial resistance and UV resistance as well. Essentials oils and powder supplements that add a noticeable favorable result include dragon’s blood, cumin oil, marine powder, sweet marjoram oil, cedarwood oil, rosehip oil, ginseng powder, petitgrain oil, kukui oil, patchouli oil, sacred mushroom powder, and cinnamon cassia oil. These additives are identified to have a significant effect in presenting a favorable outcome, in comparison to blanks, because antimicrobial zone’s of clearance have an increase to 1.5 cm or greater. With these oils and powdered supplements, we plan to test different concentration to identify the smallest ration of materials needed to get antimicrobial resistance and UV resistance while additionally testing different known pathogens that cause skin problems. Although this UV light resistance is very favorable, with regards to the marketability, we need to find a way to make samples that do not cause much deviation to natural skin color. One example of this is usually when a powdered supplement, such as marine powder, is added- marine powder has a deep green tint and with consideration to aesthetic, it would be interesting to see people walk around with green skin.

63 samples were able to demonstrate antimicrobial resistance and 75 samples demonstrated UV light protection. Of the total number of samples, 45 of them were proven to have both UV and antimicrobial protection properties. The samples that we identified to provide the best results include: RP5, RP12, RP21, RP22, RP28, RP30, RP62, RP63, RP64, RP65, RP66, RP67, RP68, RP69, RP72, RP73, RP74, RP75, RP76, RP77, RP87, RP89, RP90, RP91, RP92, RP93, RP94, RP97, RP98, RP99, RP100, RP101, RP102, RP103, RP104, RP105, RP106, RP107, RP108, RP109, RP110, RP111, RP115, RP116, and RP117. Although a majority of these samples do provide great antimicrobial resistance and UV resistance, we need to revise samples RP 63, RP64, RP67, RP68, and RP72 because they have a deep green color that may stain skin. With all samples, we plan to rework the concentration ratios in hopes of identifying if we can decrease the amount of additive materials needed to make samples pungent.

In the future, we anticipate producing more samples that prove better UV protection by the incorporation of compounds from natural powdered supplements and to actively expand our collection of essentials oils in hand based on knowledge of herbal effects demonstrated by plants. To expand on the antimicrobial effects of our samples, we hope to test against different known pathogens that cause epidermal irritation, such as Candida albicans. By the end of this project, it is anticipated that we will be able to provide a means to address more sterile wound healing techniques and a more natural alternative to UV protectants- or produce a unique cure all that reflect both results.

Blog 3: An Exotic Butter Formulation to Enhance Bacterial Resistance and UV Protection

Rudra Persaud

March 11, 2019

An Exotic Butter Formulation to Enhance Bacterial Resistance and UV Protection

Dr. Jaimelee Rizzo

My research is progressively culminating in finding the best antimicrobial formulation that will give rise to UV protection. So far I have been able to create 129 different samples- 63 samples were able to demonstrate antimicrobial resistance and 75 samples demonstrated UV light protection. Of these, 45 of them were proven to have both UV and antimicrobial protection properties.

As we continue down this research path, one question that I have is how can we lower the concentration of exotic butter, essential oil, and powder supplement combination to generate a product that will have a low risk for being toxic to humans. Although plant-based fatty acids and other metabolites do a great job at killing bacteria, at high concentration they may be harmful to human health- as the saying goes, too much of a good thing is bad. Another question/ challenge that comes up is how can we address the shelf life of these products we are creating. This question would require keeping samples stored over time and progressively testing them against bacteria every month. It is important that we can create an antimicrobial surface that has great resistance to bacteria and provide UV resistance over the course of months without the need for chemical additive/ preservative because the mission to keep everything all natural.

While we move forward with this project with these questions in mind, I am also actively seeking new essential oils that can be used to enhance my arsenal of antimicrobial surfaces to consider for moving to the production steps of this project. Some oils that have previously worked successfully to reach both outcomes include bergamot, ginger, cinnamon cassia, patchouli, kukui, petitgrain, cedar wood, rosehip, sweet marjoram, cumin, hemp, vetiver, chaulmoogra and the powder supplements that have been successful include marine powder, ginseng, and sacred mushroom. For the next wave of surfaces created, oils such as bael, cranberry seed, and calendula seem to have ideal chemical constituents that will work for both antimicrobial activity and UV resistance.

So far Dr. Rizzo has been a great mentor in helping me stay on track with research, the school year can get hectic and it is easy to lose time. Dr. Rizzo has also pushed me to present my research on many different platforms as well, in fact, I am looking forward to presenting my research in a national American Chemical Society Conference in Orlando (special thanks to the Office of Student Success for helping fund this trip!).

My biggest take away from this project is that often times research leads to more questions than answers, and because of this patience is key. Although some of our formulations did not work successfully, we are constantly trying to make them work by adding more natural products to increase their effects- but again this may be a double-edged sword because we do not want to create a toxic product.

Blog #2: Exotic Butter Formulation to Enhance Bacterial Resistance and UV Protection

During the entirety of the fall semester I have devoted time to participate in research with Dr. Jaimelee Rizzo- under Dr. Rizzo’s supervision, I synthesized antimicrobial surfaces. Our formulation of surfaces is produced via the combination of various organic butters as a base and the strategic addition of plant-based oils and powders to promote the healing effects produced by the butters.

The defining feature of this research project lies in exploring natural ways to prevent and cure bacterial infection while preventing UV damage produced by natural causes. With the understanding that Tamanu butter, Aloe Vera butter, and Ucuuba butter are known to have soothing antimicrobial effects, we wish to find chemical counterparts that are found from other organic sources that work synergistically to enhance the probability of the desired outcomes. To date I have generated about 130 samples- 62 have proven to have great antimicrobial resistance and 74 have ample UV resistance. Of all samples, 44 have demonstrated the best outcome of having both antimicrobial resistance and UV resistance.

As the primary researcher, the biggest question I have is what are ways that I can revise the samples that did not work to further enhance their effects to produce the desired epidermic outcome. By exploring the effect of adding powdered supplements such as Graviola, wheatgrass, or sacred mushroom I hope to multiply the healing and protective effects of samples. Another question that comes up during research involves understanding what other bacterial strains can be tested to ensure we are making a holistic cure to infection. So far, the bacteria we study in relation to antimicrobial resistance is E. coli, S. aureus, and P. aeruginosa.

The problem-solving methodology used to address questions like these rely on prior observations and informative literature searches to find potent organic chemicals that prevent infections and disease- by relying on past works conducted, I can stand on the shoulders of past researchers who have studied the chemical make up of plants and role that plant chemicals play in bacterial resistance.

Exotic Butter Formulation to Enhance Bacterial Resistance and UV Protection

Rudra Persaud and Dr. Jaimelee Rizzo

My research project aims to enhance the cosmetic chemistry world by introducing a novel line of antimicrobial surfaces derived from a synergetic fusion of exotic butters, plant-based essential oils, and plant-based powder supplements. Antimicrobial surfaces are any agent that has a potential to inhibit the growth and development of microorganisms- bacterial and fungus being the target group in this study. In addition to having this retardation effect, we also examine the potential for our antimicrobial surfaces in repelling the absorption of UV light. The ingredients that constitute the basis of our formulation are ucuuba butter, aloe vera butter, and tamanu butter; the variable ingredients include a wide variety of oils and powders including, but not limited to, chaulmoogra oil, Jamaican black castor oil, dragon’s blood, marine phytoplankton, and sacred mushroom. The products that we generate in our lab are considered vegan and 100% organic- one of the biggest trends found in the cosmetology market is a global shift for consumers to favor products that are derived naturally instead of synthetically. In general, natural skin products are earth-friendly, biodegradable, reduce any uncomfortable chemical irritation, are nutrient rich with natural fatty acids, and prevent adverse chemical imbalances. The overall premise of this project is to provide more natural means to sterile wound healing techniques and prevent harmful cancers caused by UV rays.

The antimicrobial surfaces are synthesized and tested for UV resistance in our research lab at Pace University, but to test for antimicrobial resistance our samples are sent to a partner university, Long Island University Post.

To generate the antimicrobial surface, the individual butters used are initially liquefied through the use of a hot water bath. About 4 mL of each sample is aliquoted into a beaker and 2 mL of an essential oil is infused. The mixture is then plated in a petri dish and left to solidify at room temperature.

Using a tryptic soy agar plate, 0.8 cm plugs of agar were removed. 175 µL of each butter and oil sample was liquefied and placed in different agar holes, which were left to be hardened. 100 µL of 106 S. aureus was spread on the agar using a sterile spreader and each plate was left to incubate overnight at 37 °C. Antimicrobial activity was assessed based on if the zones of clearance were greater than 1 cm around each sample plug.

Following the creation of the surface, UV radiation tests are conducted. Each antimicrobial formulation is generated using the above procedure. A ziplock bag is filled with UV detecting color beads, and each antimicrobial surface is applied evenly over the bag. Each bag is then placed under UV radiation for about 20 seconds. Samples were then rated based on how drastic the color change was in relation to a blank sample of individual butters and a comparative sunscreen.

Ucuuba, Tamanu, and Aloe Vera Butter As A New Formulation to Enhance Bacterial Resistance And UV Protection- Final Report

In the past 3 months, I have been working diligently under my mentor, Dr. Jaimelee Rizzo, to investigate various ways to synthesize natural antimicrobial surfaces that provide protection from UV rays. Antimicrobial surfaces have the potential to prevent the growth of microorganisms, i.e. bacteria. The recipe for my surfaces is a combination of exotic butters, natural plant-based oils, and powdered supplements. In my experiments, the fixed dependent variable is ucuuba butter, aloe vera butter, and tamanu butter and the independent variables are the different plant-based oils and powdered supplements-which are infused to enhance the properties of the butter. Naturally, the butters have known restorative properties that aboriginal civilizations used for centuries to help with varying dermatological conditions; by infusing different oils and powdered supplements, we can fortify these butters to have increased wellness abilities. The antidotal effect of our starting materials is attributed to fatty acids naturally produced in their respective flowering plants. Various studies suggest essential fatty acids and their metabolites are effective in reducing the epidermal burdens caused by microbes. Common small chain fatty acids found in the starting material are trimyristin, lactins, salicylic acid, glycerols, and glycolipids- these are known to inhibit the growth of bacteria. Inherently these biological constituents are the first line defense in warding off infections in the body.

Using all-natural ingredients is favorable in this project because the skincare industry is straying away from the use of chemical products. In general, natural skin products are earth-friendly, biodegradable, reduce any uncomfortable chemical irritation, nutrient-rich with natural fatty acids, and can prevent adverse chemical imbalances throughout the body. In concluding this project, it is anticipated that we will be able to provide a naturopathic remedy for sterile wound healing while working to prevent sun-induced cancer.The formulation of antimicrobial surfaces can be divided into two protocols: the first is to generate and test the antimicrobial surfaces followed by demonstrating ample resistance towards UV radiation.

To being, individual butters are liquefied through via a hot water bath. About 4 mL of each sample butter is aliquoted into separate beakers followed by the addition of 2 mL of an essential oil is infused, and no more than 1 gram of powdered supplement. The mixture is then plated in a petri dish and left to solidify at room temperature. These samples are then transported to a partner university, LIU Post, to be tested for bacterial resistance.

Using a tryptic soy agar plate, 0.8 cm plugs of agar were removed. 175 µL of each surface was liquefied and placed in different agar holes, which were left to solidify again. 100 µL of 106 S. aureus was spread on the agar using a sterile spreader and each plate was left to incubate overnight at 37 °C. Antimicrobial activity was assessed the next day based on the diameter with which bacteria did not grow around the antimicrobial surface- this area is considered the zone of clearance, where a diameter greater than 1 cm is classified as antimicrobial.

In a transparent ziplock bag filled with UV detecting color beads, each antimicrobial surface is applied evenly over the bag- in a manner similar to the application of sunscreen. UV radiation is then induced over each ziplock bag for about 20 seconds. Samples were then ranked on a 1-10 scale based on the drastic color change in relation to a blank sample of individual butters and a comparative sunscreen.

So far, 14 different essential oils were incorporated in each of the three butters. Overall, 94 different samples were concocted, and about 45 different samples provided ample microbial resistance when tested against S. aureus. When tested for UV protection, about 40 samples demonstrated some degree of UV protection. About 35 samples were able to demonstrate both UV protection and antimicrobial resistance. The materials most commonly combined to get desirable results incorporate black cumin oil, marine powder, and ginseng powder. The most favorable results recorded to date are samples RP 65-69, 73-77, 83-85. Specifically, RP 65 had the widest margin to inhibit bacterial growth at greater than 4 cm and no UV light penetration- this sample was made with a mixture of 3.85 mL of ucuuba butter, 3 mL of black cumin oil, and 0.2 grams of marine powder. Looking forward to the continuation of this project I hope to discover more combinations that offer a zone of clearance greater than 4 cm while advancing this study to test our samples on live living cells.

As I am to graduate in spring 2019, weighing my options for a career is a pressing matter. Having spent time in Dr. Rizzo’s lab studying a cosmetic chemistry based project, this seems like a potential field of work. Understanding that the hardest part of research is failure and that patience in making samples work is the only drawback; striving to help out others is the reward and motivating force. Having Dr. Rizzo as a mentor is really beneficial because she gives me enough space to explore my research personally in order to figure out what works and what doesn’t, but she is opened enough where she welcomes questions to further my understanding of what is happening in the lab.

Components and Results -1vu56hy

 

Ucuuba, Tamanu, and Aloe Vera Butter As A New Formulation to Enhance Bacterial Resistance And UV Protection: Blog 2

Throughout the course of the summer, I have been conducting research, under the supervision of Dr. Jaimelee Rizzo, where I have studied ways to generate antimicrobial surfaces that can provide UV resistance by combining indigenous kinds of butter, natural oils, and organic powdered ingredients. Since the beginning of this investigative study, the base of my formulations has been ucuuba butter, aloe vera butter and tamanu butter. These kinds of butter, individually, are known for having a degree of resistance to bacterial growth, cleansing properties, and rejuvenating ability which is due to the relative amounts of fatty acids naturally found in the plants they are derived from. Various studies suggest the use of essential fatty acids and their metabolites are effective in reducing the epidermal burdens caused by microbes. Small chain fatty acids common in the starting materials used in this study include trimyristin, lactins, salicylic acid, glycerols, and glycolipids, which are known to inhibit the growth of bacteria. Our bodies also naturally produce these biological chemicals to ward off and protect us from infection. The incorporation of natural oils in the production of antimicrobial surfaces further enhances microbial resistance as well as provides a potential means to obstruct UV penetration- ultimately prevent the development of cancer.

A partnered group at LIU Post, students under the supervision of Dr. Karin Melkonian, follows a protocol for the bacterial testing done on the samples generated in my experiments. At LIU post, our research associates test the bacteria S.aureus against samples and measure the diameter for which bacteria does not grow. We assess samples as being antimicrobial when the diameter of clearance is greater than 1 cm, however, we are looking for products with a far great zone of clearance, which will be tested further against other bacteria. Throughout this project, 14 different essential oils were incorporated with each of the three kinds of butter. Overall 94 different samples were concocted, and about 45 different samples provided ample microbial resistance when tested against S. aureus. When tested for UV protection, about 40 samples demonstrated some degree of UV protection. UV protection is determined based on a 1-10 scale where the degree of color change is visible on UV color changing beads, 10 being no UV light penetration. Overall, about 35 samples were able to demonstrate both UV protection and antimicrobial resistance. In general, the materials that are most commonly combined to get the results that we consider most favorable are samples that incorporate black cumin oil, marine powder, and ginseng powder. The most favorable results that have been recorded today are samples RP 65 which had the widest margin to inhibit bacterial growth at greater than 4 cm and absolutely no UV light penetration- this sample was made with a mixture of 3.85 milliliters of ucuuba butter, 3 milliliters of black cumin oil, and 0.2 grams of marine powder. Looking forward in the continuation of this project I hope to find more combinations that offer a zone of clearance greater than 4 cm and advancing this study to test our samples on live living cells. By the end of this project, it is anticipated that we will be able to provide a means to address more sterile wound healing techniques.

Looking to the future, as I am projected to graduate in spring 2019, weighing my options for a career is a pressing matter. Having spent a quality amount of time in Dr. Rizzo’s lab studying a project closely related to cosmetic chemistry, this seems like a potential field of work. Understanding that the hardest part of any research project is failure and that patience in finding samples that work is the only drawback to this type of study, but striving to help out a population of others is the potential reward and driving force. This research project has the potential to change the way wounds are treated in active environments. Although chemically enhanced skin care products, which are used daily, are effective they carry a lot of side effects and some unknown long-term changes may be observed. Switching to natural products is a safe way to treat wounds or infections while preserving a natural chemical balance of the human body.

 

Investigative Questions:

What are different ways to increase the zones of clearance of the tested surfaces, which will increase antimicrobial resistance?

How can we maximize UV resistance and antimicrobial growth concurrently while maintaining the smallest possible ratio of materials?

 

Components and Results -1vu56hy

“Ucuuba, Tamanu, and Aloe Vera Butter As A New Formulation to Enhance Bacterial Resistance And UV Protection” Blog Post

The research project that I have been working on over this summer encapsulates the world of cosmetic chemistry, specifically, I am studying how to generate various types of all natural antimicrobial surfaces through infusing exotic butters and various types of oils. An antimicrobial surface is an agent that has the potential to prevent the growth of microorganisms, such as bacteria; concurrently in the lab, ultraviolet tests are done to determine if these surfaces will also provide resistance to UV light. For these experiments, the fixed variable is the type of base butter that is used- ucuuba butter, aloe vera butter, and tamanu butter- whereas different plant-based oils are combined with these butters to enhance the healing and protective characteristics of the butter. One of the reasons why the use of all natural ingredients is favorable in this project is due to the fact that the skin care industry, typically high-end companies, is straying away from chemical products. In general, natural skin products are earth-friendly, biodegradable, reduce any uncomfortable chemical irritation, nutrient-rich with natural fatty acids, and prevent adverse chemical imbalances. By the end of this project, it is anticipated that we will be able to provide a more natural means to sterile wound healing techniques and prevent harmful cancers caused by UV rays.

The methods of this project can be broken up into two parts: the protocol to generate the antimicrobial surfaces and the protocol for the UV radiation test.

To generate the antimicrobial surface, the individual kinds of butter used are initially liquefied through the use of a hot water bath. About 4 mL of each sample is aliquoted into a beaker and 2 mL of an essential oil is infused. The mixture is then plated in a petri dish and left to solidify at room temperature.

Using a tryptic soy agar plate, 0.8 cm plugs of agar were removed. 175 µL of each butter and oil sample was liquefied and placed in different agar holes, which were left to be hardened. 100 µL of 106 S. aureus was spread on the agar using a sterile spreader and each plate was left to incubate overnight at 37 °C. Antimicrobial activity was assessed based on if the zones of clearance were greater than 1 cm around each sample plug.

Following the creation of the surface, UV radiation tests are conducted. Each antimicrobial formulation is generated using the above procedure. A ziplock bag is filled with UV detecting color beads, and each antimicrobial surface is applied evenly over the bag. Each bag is then placed under UV radiation for about 20 seconds. Samples were then rated based on how drastic the color change in relation to a blank sample of individual kinds of butter and a comparative sunscreen.