Third blog Spring 2011

 

Biomind program provided me the opportunity to develops fundamental research techniques. This competitive program definitively was a helpful one, because allows me to be involved in a scientific world. Also through out this program I have the opportunity to participate in different symposiums doing oral and poster presentation and this experience enhances my scientific knowledge and help me to known expertise people in my research field. This is a very important thing for my future because it work as a motivation and model to me as a scientific. Is important to mention that thanks to this program I have the opportunity to have a close interaction with my mentor, which allows me to get advise and advantage of her knowledge. As part of my scientific skills, this program help me to develop my writing, speaking, and reading. One of the most important benefits that I get from this program was the opportunity to have a good overview about scientific research world, the requirements to be a good researcher and the importance of the investigations for society development. Finally, Biomind program had big influence in my decision to continue graduate school, because it allows me to adquire and expand my scientific knowledge and techniques and motivates me to want to persuade the PhD degree to have the opportunity to conduct research independently.

To see the collage please check the following link: http://researchadventures.blogspot.com/    

If you have any problem to access the link be free to contact me.

April 5, 2011 at 1:25 pm Leave a comment

2011 Spring Second Bioblog

 

      During this semester I wasn’t able to obtain critical results because the principal instrument to analyze the data is not function (HPLC). However I prepared several trials for this analysis so I’m waiting to fix the equipment to get several important data I hope. However I’m working in other research route as results of data obtained last semester. Specifically,I developed a soxhlet extraction to the momordica charantia leaves using methanol as a solvent. The extract obtained was subjected to a column chromatography and in this moment I’m grouping the fractions obtained for further identification HPLC, NMR and UV analysis. In other part I’m performing an Artemia salina bioassay to the fractions recovered last semester from Preparative TLC analysis and water soxhlet extract column chromatography. Some of the fractions obtained in the previous analysis mentioned presented consistent signals in NMR with compounds reported in the literature that had presented biological activity. Having this in mind I decided to perform an Artemia Salina Bioassay.

     In this moment I’m at the middle stage of my semester goals even I can’t complete the data analysis because the sample is prepare and I’m waiting until the equipment will be fixed. So I can said that this one obstacle that I had this semester and to overcome it, I continued preparing sample and following other research route to be sure to maintain obtaining result. I get some trouble to perform the Artemia Salina Bioassay following the protocol using by other researcher here in UPR Cayey. For example I found that in the negative control where all the shrimp are suppose to be alive, they died. To overcome that I decide to reduce the growing time of the shrimp (was 2 day and now I’m doing the analysis at 1.5 days). Right now I’m waiting for results. I’m sure that in the next month I will have significance results.

March 8, 2011 at 11:09 pm Leave a comment

First Blog Spring 2011

Isolation and Identification of Potentially Bioactive Compounds from Momordica
charantia L. “Cundeamor”

 

Momordica charantia L. locally known as “Cundeamor” has been identified as natural medicinal source for diabetes treatment. According to regional claims, the tea preparation from the leaves is used for reduction blood glucose level. Moreover a recent literature review revealed that species of this plant has been extensively studied, reporting the characterization of compounds with potential hypoglycemic activity from the leaves and fruit. However, in Puerto Rico “Cundeamor” has not been widely studied. Recognizing the variation of physical appearance of this plant along the countries, the overall chemical composition should have active metabolites. The purpose of this research is to validate the hypoglycemic of isolated compounds from Puerto Rico “cundeamor” species infusion and Soxhlet extracts. Currently, this research is in the middle stage. Preliminary spectroscopic results showed comparative and consistent signals in 1H NMR analysis from dichloromethane infusion and Soxhlet extracts with active compounds previously reported in the literature. As part of the methodology an infusion was performed in order to simulate homemade tea preparation and a Soxhlet extraction with dichloromethane and water respectively, were carried out to obtain a crude extract. Column chromatography and HPLC analysis were used as purification techniques. Qualitative, quantitative and spectroscopic results obtained from infusion and Soxhlet extracts will be presented.

February 3, 2011 at 1:58 pm Leave a comment

Third Blog (Fall 2010)

   Working in the Laboratory

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            During this semester I’m developing different analysis from the plant Momordica charantia, commonly used to treat Diabetes Mellitus type II according to popular claims. These analyses include quantitative, spectroscopic and chromatographic ones.  The extraction process in the infusion analyze was optimize, and the results revealed that more extraction with less solvent allow to recover the same or higher amount of extract. The maximum yield recovered was 0.048%.  The TLC analysis was optimized, founding a mobile phase that allowed a better separation of the components in the infusion and soxhlet extracts, TLC preparative fractions. This mobile phase was the mixture of Hexane: Ethyl Acetate: Methanol (6:3:1).   Additional, spectroscopic analysis were performed and the 1H NMR spectrum revealed comparative signals between the extract recovered from the infusion using dichloromethane as a solvent and soxhlet analysis using water following by dichloromethane liquid-liquid extractions. Also these spectrums present consistent signals with compounds reported in the literature which a hypoglycemic effect was validated. Then, UV-Visible analysis of the Preparative TLC fractions has been revealed that even theses fractions were obtained from each signal in the TLC analysis, various absorption wavelength was founded from each fraction. According to this spectroscopic analysis the fraction 4 of the preparative TLC is the less complex, presenting 2 important absorption bands. Finally using an stationary C18 columns and H2O:Methanol (1:1) mobile phase a HPLC analysis was carried out for the fraction 4 of the preparative TLC.  The results obtained showed four trace signals, but the most abundance was obtained at 5.82 minutes and it is close to the 100% peak area.

November 22, 2010 at 6:53 pm 1 comment

Fall 2010 Second Blog

Saraí Quiles

Research Topic

Study of characteristics of different algae that will contributes for rivers contamination.

According to this research, algae are involves in the river contamination. The purpose of this research is to determine the mechanism or properties of the algae that contribute for the environment pollution. This investigation involves the use of different important techniques. The microscope is used for the identification of different algae culture and physical properties of each one.  In other part the isolation of algae include complicated procedures that involve the use of chromatography techniques. This semester the research focus include the identification of the 5 most abundance algae in the rivers and to study the growing and environmental resistant properties. 

I really enjoy to learn about this kind of research because it is related directly to understand or solve some nature issues that have several impact in a society. I’m very sure that this research will allow to Saraí to develop new skills or techniques.

Priscila Rodríguez

Research topic

This research is conducted in order to build  E. coli clone’s library.

When Priscila started in Biomind Program she was working in a multidisciplinary research which involved molecular biology and electrochemistry. The main purpose of this research was to study Geobacteraceae microorganisms. This research was focus to understand the microbe-electrode interaction in order to increase the power output of these microbial fuel cells. This investigation allows acquiring some molecular biology techniques such as bacteria’s plasmid clones and other technique that allows the isolation of a specific species of Geobacteraceae. This semester she has the opportunities to practice some previous techniques such as bacteria’s plasmid clones, plasmid extractions and insertion on E. coli bacteria and to develop and understand the importance of PCR technique.

I think that the change of research definitively is a challenge that Priscila accepts, but this opportunity will result in the acquisition of new techniques and improving of research experience. E. coli bacteria is a versatile organism that will help to study and discover some DNA information or techniques important to help to solve some issues of the society.

October 23, 2010 at 1:41 am 1 comment

Fall 2010 First Entry

2010 Summer internship

 During this summer I had the opportunity to make research in Department of Chemistry on Penn State University. There, I was working in a research entitled: Stopped flow studies of Compound I in Cytochrome P450 CYP119. Cytochrome P450s are heme proteins involves in hydroxylation of unactived C-H bonds in many organisms. These proteins has pharmaceutical interest because can allows develop new pharmaceutics and the optimization of known drugs production. The study of two intermediates identified in the catalytic cycle of these proteins (Compound I and II), is the focus of Dr. Green’s laboratory. My research was conducted in order to study Compound I in a thermostable Cytochrome CYP119. Determination of kinetic rate constants involved in the formation and decay process of Compound I and the optimal conditions to produce high yield of this intermediate in P450 CYP119 were my contribution for P450 studies.

This REU summer experience was great to know more about graduate studies in chemistry. This research involved multiple disciplines such as Physical, Inorganic and Biochemistry, allowing me to learned many new techniques. Some of these are DNA extraction, cells transformation, proteins expression in E. coli. Also, I learned how to purify three different proteins (P450BM3, CYP119 and Chloroperoxidase), including how to use the Fast Protein Liquid Chromatography and BIO-RAD instruments. I investigated formation of Compound I using Stopped flow and GC-MS. To analyze this data, I learned how to use Igor Pro and Pro-Kineticist programs. This research has been extremely helpful to increase my research experiences and I know that will help me a lot to achieve one of the most important goals of my life, earn a PhD in Chemistry.

Research purpose for fall 2010

For this semester I have many objective that I will achieve:

  1. I will develop effectively TLC analysis for the infusion extract.
  2. Also I will identify and characterize some compounds from the complex momordica charantia crude extract using the Preparative TLC analysis in order to simplify the extract, the HPLC analysis as the principal technique for optimum purification process and finally spectroscopy techniques such as UV and NMR for the characterization of purified compounds.
  3. I will identify and characterize some compounds from the infusion extract using the HPLC analysis for optimum purification process and spectroscopy techniques such as UV and NMR for the characterization of purified compounds.
  4. In addition I will obtain comparative data from the infusion and crude extract with data reported in the literature.
  5. Finally I will obtain reproducible data in term of the recovery percent from the infusion and soxhlet extract with data previously reported.

This semester is a continuation of the last one. All the results obtained last semester has been helpful to plan this semester. The comparative results obtained last semester in the NMR of infusion of soxhlet extract, help us to known that the extraction procedure was effective and this semester I will continue using it. Also I will continue analyzing the fraction obtained in the column chromatography column last semester. As I said before the progress done last semester allows me to be close to achieve one of the principal goal of this research.

During this semester I will using some new techniques:

  1. Preparative TLC: used to simplify the complex sample obtained from the soxhlet extraction of Momordica charantia leaves. 
  2. HPLC: used to separate the compound in the extract using polarity properties.
  3. UV: used to monitoring the extraction procedure.

September 23, 2010 at 2:14 am 1 comment

2010 Third Bio-blog

During the visitation of three blogs of biomind researcher I find: 

Nahira Arocho

 Nahira Arocho new research sounds interesting but in my particular case I don’t understand the purpose of the research. I know that is related with zoologist field research, but maybe in the future after be more engaged  with the research she can explain about the goal of the research and the specific objective. My perception is that she is now working in the adaptation process of this new research. In the future probably am going to look again the blog.

Rigoberto de Jesus

Rigoberto de Jesús biochemistry research is so interesting, because a lot of researchers are working in the production of bioethanol, but this specific procedure sound different. Work with enzymes revealed that he is working arduous. Also is more difficult optimize the ethanol production finding the better enzyme-bioreactor interaction. Hopefully in the future his research will help for the production of ethanol reducing the environmental pollution.

Lilliana Emmanuelli

 Recently a lot of researchers conducted their research to biomedical goals. That is good because a lot of medical diseases can be understood and a treatment can be obtained. Lilliana’s neurosciences research is so interesting and I know that in the way as the research is conducted positive data will be obtained. I hope that she can continue working so hard to get her research team goals and help the society.

Research Work

This semester was very successful in comparison with the last one. Last semester I was learning apply the techniques acquired in Organic Laboratory to a macroscale experiment in the research.  During this semester I don’t learned a new technique but I practice and optimize the techniques learned last semester. In other part I learned more about compounds presented in the infusion and soxhlet extract using the thin layer chromatography analysis. Also I learned that an arduous work giving to you a satisfaction, when you can obtain positive data.

                This semester I had some challenges that I overcome working hard and applying my knowledge about the research techniques and literature review information. One of this challenge was the process of obtain reproducible data with previous results. In this case it wasn’t so hard, but I had to work hard to do the same work as previous researcher in my investigation. Other challenge was tried to obtain positive results in the artemia salina bioassay, because in the literature several papers reported bioactivities presented of compounds extracted from Momordica charantia in others countries. However wasn’t possible obtain positive results, but applying the knowledge about the plant and the extract was possible explain the negative results obtained in this analysis.

So this analysis let us to keep the artemia salina bioassay out of this investigation and just get focus in the identification and characterization of the compounds obtained in the infusion and in the soxhlet extract and compare the results between them and with the bioactive compounds reported in the literature. So for the next semester I have a more arduous work in the process to characterize the compounds purifying it using the Column Chromatography and High Performance Liquid Chromatography (HPLC) analysis.

April 18, 2010 at 11:03 pm Leave a comment

Jan-May 2010 Second Entry

        During my second year as undergraduate student I learn an interesting technique in my Organic class used to identify and compare compounds; that it the Thin Layer Chromatography. This is a cheaper technique that does not need more than a Silica Gel plate (stationary phase), and a solvent mixture (mobile phase). In my research this technique is highly used as a way to get a technique to identify and compare the sample obtained after the extraction, between previously reported. Also this technique allows me to monitoring the extraction process knowing if the interested compound is present in the sample. In my research this technique is very useful, in order to obtain reproducible data, several times the sample obtained after the infusion is subjected to this technique to compare the results with previously reported. This allows me be sure that the compounds extracted are the same as reported previously, that presented an interesting signals in the 1H NMR.

      In this moment I can say that my research purpose for this semester is a 3, define as intermediate progress. I did two trials of the infusion extractions, and Soxhlet extraction with Dichloromethane. Also, I try to identify the sample obtained after the extraction process with comparative TLC, and NMR analysis was performance. The Soxhlet extraction with methanol has not been performance yet and some struggle with the Artemia Salina Bioassay doesn’t allow finishing the semester working purpose. So I’m still working in it, to be sure to achieve my goals for this semester. In the infusion extraction process, at the first time made during this semester the recovered percent wasn’t enough to performance an Artemia Salina Bioassay. In order to overcome it another infusion extraction was carry out more carefully to be sure that enough sample would be recovered. Other difficulty occurs in the process to performance the Artemia Salina Bioassay. In this analysis the positive control doesn’t work well, so this analysis was carry out two times, but it still presenting difficulty.

March 7, 2010 at 1:11 am 1 comment

2010 First Entry

      During this semester I will be repeating again the infusion and soxhlet extraction with the goals to optimize different analysis that the past semester doesn’t revealed satisfactory results. One of my objectives for this semester is optimize the TLC mobile phase solvent ratio to have a successfully separation of the compounds obtained in the sample from the “Momordica charantia” leaves. The second objective is optimizing the Artemia Salina bioassay procedure to obtain reproducible results. In order to achieve that I’m going to follows an accurate procedure founded in the literature. And the last but not less important objective is the identification of the compounds recovered in the sample by the Hydrogen Nuclear Magnetic Resonance (NMR) analysis.

       These objectives are highly related with the work realized during the last semester. The first things that establish the relation of these objectives are the different optimization processes of the analysis performance during the past semester. Also we are following a common investigation steps, where the first thing is isolated the compound from the “Momordica charantia” leaves, then use an different characterization analysis of the compounds obtained and finally the identification of those compounds.

     Throughout this semester I will be expose to strengthen different research skills such as working with accuracy, organization and good research development; use of literature, develops and infusion and soxhlet extraction, use of the computer in the NMR analysis, work in a macro scale and others.

February 4, 2010 at 10:09 pm 1 comment

Third Entry

      Ariel González’s research is about an interesting topic. He is study the Ostoblastic cell and how can these cells grown over different surfaces, like Ti-6Al-4V, this is a metal conveniently used in the aerospatiale equipment for its mechanical properties. Ariel is already prepared to apply some techniques like electrophoresis and DNA amplification; he did a great job.

     Angela Alicea‘s research topic is so interested to the immunology and virology fields. This research is about the antibiotic resistance of bacteria, and how can be altered the genes sequences. Also mention that bacteria can be antibiotic resistance without been exposed to a antibiotic. This research causes a curiosity to me because I never had thought that this can be possible. In this research are studying the E.coli bacteria using Tetracycline antibiotic to induce the antibiotic resistant in the bacteria.

        Nydia Burgo’s research topic is helpful to the humanity health. This research is about the cause of cardiovascular diseases such hypertension, arrhythmia, etc. Her work team wants to know more about how can, the variation on different ions concentration like calcium (Ca) can provoke some cardiovascular disease. Their future goal is create a drug that can remediate some cardiovascular diseases.

        During this semester I really learned about my research topic. The literature review helped me in this process to know the biological activities of compounds presents in the Momordica charantia plant reported in different investigation (antioxidant-anti-VIH, antidiabetic, antiulcer, etc). Also with this process I learn about different extraction methodology used to isolate biological active compounds as triterpenoids and phenolic compounds from this plant. In the extraction process of Momordica charantia leaves using the infusion technique the first challenge was the recuperation of compound after finish the infusion extraction process. Low quantity of sample were recovered because that was my first extraction process using a macro scale and I didn’t have enough aware about the process. In order to solve it I did again the extraction process more carefully and the extract quantity was higher than the first time. Also we have some struggle with the artemia salina bioassay because in this test wasn’t possible the shrimp growing following the protocols established. In order to overcome it, we were checking the different variable that can be affected the shrimp growing. After several trials we concluded that the problems wasn’t the methodology but was that the shrimp eggs weren’t in good condition to growing (they had some damage).

     Right now, I’m in the process of do the Thin Layer Chromatography analysis to the infusion extract and soxhlet extract. I hope finish at least the artemia salina bioassay of the infusion extract and soxhlet extract and can characterized those extract by Proton (1H) and carbon (13C) Nuclear Magnetic Resonance. For the next semester our research goals will be made a HPLC analysis from the extract recovered in the infusion and soxhlet techniques, make a more specific biological test to know the antidiabetic activity of those extract and finally try to make a reproducible infusion extraction and characterized those extracts.

November 15, 2009 at 2:08 pm 1 comment

Older Posts


Recent Comments

Rosa Buxeda on Third Blog (Fall 2010)
Rosa Buxeda on Fall 2010 Second Blog
Rosa Buxeda on Fall 2010 First Entry
Rosa Buxeda on Jan-May 2010 Second Entry
Rosa Buxeda on 2010 First Entry
Rosa Buxeda on Third Entry
Rosa Buxeda on Second Assingment
Rosa Buxeda on First Assignment
Mr WordPress on Hello world!

Calendar

January 2021
M T W T F S S
 123
45678910
11121314151617
18192021222324
25262728293031