Valentine is still so far away from now, but I am feeling my world is so pink. I don’t know what happened to me that currently I am being crazy in pinky-pinky. Look at that! I bought a pinky mirror, either my body wash, detergent, my hand phone and CD holder. What so cute and romantic they are. Am I romantic guy?? I think so, he3x……
Last time, pink was favorable by ladies but nowadays, when we go to the mall, café, and so many places, we will find out, a gentleman also has been confident wearing a shirt, tie, or bringing laptop with the pink color. No need to be shy and just be confident with yourselves that if you feel comfortable with anything you wore, so does people look at you.
So what??? Actually, I just finished one part of my research. As we knew that I had been successfully synthesizing five new compounds which was classified as benzilidene ketone derivatives. Based on the structure-activity relationships, that such related compounds shown the antioxidant activity. What is that?? And the story goes…..
From the word ANTI (=against) and OXIDANT (=any chemical which cause oxidation process). Oxidation is the process in which 2 chemicals having action to donate-accept electron one to each other. Normally, this kind reaction is required to conduct some biochemical process such as combustion, oxidative phosphorylation, conjugation and so forth. But, in an extremely high energy, this reaction is becoming uncontrolled and triggering some further reaction that impact to cell damages.
Combustion need O2 as a fuel
In the normal condition, the donating-accepting electron will be occurred in pair form (pair electron) what we call as heterolytic reaction while an extremely high energy will INITIATE the reaction to be homolytic one (singlet electron). God always create everything in the world to be a couple; big-small, rich-poor, boys-girls, etc, so when anything is in single it seemed like not too stable and tendentious to be looking for the couple.
Homolytic & Heterolytic process in forming free radicals
So does the electron, when it is in singlet state, it will become RADICAL. This free radical will wildly attack anything to get a doublet state. Worst, the high energy will spread out some propagation reaction (chain reactions) that expands the target as large as possible. The simplest example, our body which is composed by organic compounds (polysaccharide, protein, lipid and nucleic acid) will be easily exposed by free radical especially when it is not in a fit condition.
Free radical resources can be derived from pollutant (industrial waste, uncompletely combustion, fume), short wave UV light exposure, chemicals (preservatives, coloring agent), etc. Usually, we never realized when we contaminated which those matters. The reaction is taking a time and chronically so our body will responds when the product of reaction has been accumulated. Once the radical attacks our cell’s body, some metabolic pathways will modulate some organ function disorders and it causes degenerative diseases such as Diabetic Mellitus, cancer, and aging progress.
To protect the cells and organ systems of the body against reactive oxygen species, humans have evolved a highly sophisticated and complex antioxidant protection system. It involves a variety of components, both endogenous and exogenous in origin, that function interactively and synergistically to neutralize free radicals.These components include:
1. Nutrient-derived antioxidants like ascorbic acid (vitamin C), tocopherols and tocotrienols (vitamin E), carotenoids, and other low molecular weight compounds such as glutathione and lipoic acid.
structure of vitamin C
structure of tocopherol
2. Antioxidant enzymes, e.g., superoxide dismutase, glutathione peroxidase, and glutathione reductase, which catalyze free radical quenching reactions.
3. Metal binding proteins, such as ferritin, lactoferrin, albumin, and ceruloplasmin that sequester free iron and copper ions that are capable of catalyzing oxidative reactions.
4. Numerous other antioxidant phytonutrients present in a wide variety of plant foods.
In recent years, epidemiological studies show that consumption of food with high phenolic content correlates with decreasing cardiovascular diseases. Phenolic compounds may produce their beneficial effect by scavenging free radicals. There has been much researchs which showed the implication of oxidative and free radical in the mediated reaction on the degenerative processes related to aging and other diseases.Several methods, both in vivo and in vitro, have been developed to measure antioxidant performance.
These methods focus on different mechanisms of antioxidant including scavenging of oxygen and hydroxyl radicals,5 reduction of lipid peroxyl radical, inhibition of lipid peroxidation or chelation of metal ions. Thus, some methods that are based on the mechanisms include β-carotene bleaching method,6 DPPH assay, thiobarbituric acid reactive substance (TBARS) method, lipid peroxidation, and deoxyribose assay.
deoxyribose degradation by oxidant will form malonaldehyde as product (MDA)
The MDA product can be detected by reacting with Thiobarbituric Acid (TBA) as apink color solution
Theoretically, free radical will be formed if a covalent bond happens to break. The compound which is scavenging hydroxyl radical can decrease deoxyribose degradation. Deoxyribose degradation will produce malonaldehyde that is identified by PINK color of the thiobarbituric acid (TBA) complex. Synthesis of Hydroxyl Radical Scavengers. Deoxyribose degradation is assumed as a our DNA damages when the radical attacak our cell’s body. Remember that DNA stands for Deoxyribonucleic acid.
mixing of D-ribose + Phosphat Buffer pH 7.4 + H2O2 + FeSO4, no color changing before reacting with TBA
A concentration seri of my MH1 will be tested as antioxidant
boiling will speed up the complex reaction of MDA-TBA
Done!!! My”pinky-pinky” color was formed while the orange one shown the inhibition of my MH1 toward oxidation process
My compound (MH1) which is classified as benzylidene ketone derivatives has a conjugated system and is expected to be easily oxidized. The more the double bond, the easier it will be oxidized. Therefore, it is assumed that benzilidene ketone and its derivatives will show antioxidant activity. Therefore, the objectives of this study are to develop an oxidation system using deoxyribose assay and to determine the IC50 value of each of the antioxidant.
The skeleton of my MH1, the long conjugated double bond having antioxidant effect
The method is based on the determination of MDA , a degradation product of 2-DR, by measurement of the condensation product with TBA. Typical reactions were started by the addition of either Fe(II) or Fe(III) to solutions (0.5 mL final volume) containing 5 mM 2-DR, 10 mM phosphate buffer (pH 7.2), H2O2 (peroxide was omitted in the Fe(II) autoxidation assays) and the synthetic compound (50 ppm, 100 ppm, 250 ppm, 500 ppm, 1000 ppm). Reactions were carried out for 10 min at room temperature (24–25 °C) and stopped by the addition of 0.5 mL 4% phosphoric acid v/v followed by the addition of 0.5 mL TBA solution. After boiling for 15 min, solutions were allowed to cool to room temperature, and the absorbance was measured at 532 nm.
The above method actually is the most optimized result I did since no satisfactorily PINK color produced at my previous methods. What a happy I was when the PINKY shown the FASHION. How cute it was, he3x…..
I love “pinky” like a crazy
The result shown that my compound inhibited superoxide radical with a 62.02% inhibition at 500 ppm. It was no radical responding at 50 ppm and the inhibition was getting started when the concentration was increased. Unfortunately, the radical scavenging was decreasing when the concentration of the sample at the highest one. It might be caused by the solubility of compound is getting lower at 1000ppm, in is assigned by a bit precipitation occurred during heating process.
Last but not least, my MH1 was successfully having antioxidant property and it will be a candidate for IN VIVO examination. So, when you are targeted by THE FREE RADICALS, just call me “Mr. PINKY-PINKY!!!!” …..and I will be your ‘GUARDIAN ANGEL’ to scavenge that suck PREDATORS!!!!