Fluorescein dye has been used for a very long time and it serves many purposes within the lab and in everyday life. As a product fluorescein appears to be a brownish-orange powder solid that appears to be bright, neon green under a UV lamp. Fluorescein, Yellow No.7 dye, is used as a color addictive to food, medications, and beauty products.2 It is also used as a fluorescent tag for labeling antibodies that will be targeting specific areas of a cell1 or identify and locate life threating thrombotic emboli.4 Green Chemistry is being used in order to prevent the creation of waste products. Creating less waste will have a positive effect on the environment. Also running the reaction using less harmful solvents or potentially no solvents at all.2 …show more content…
A proton transfer occurs causing the formation of water leaving group and this water leaving group is kicked off by a second resorcinol equivalent. In the second Friedel-Crafts resorcinol attacks a protonated carbonyl in order to form a tetrahedral intermediate. The lone pair found on the oxygen of the newly attached resorcinol attacks the carbon of a protonated carbonyl. This forms an ether ring. A second proton transfer occurs causing water to leave. The molecule requires a final step in which it undergoes an intermolecular Fischer esterification leading to the fluorescein final product. Analysis via melting point, UV/Vis spectroscopy, 60 MHz 1H NMR, and 400 MHz 1H NMR were performed to ensure the formation of the fluorescein …show more content…
Resorcinol (153mg) and phthalic anhydride (100mg) were added to a large test tube and placed below sand level. 3 drops of 8M sulfuric acid were added to test tube and reaction was monitored for 30 minutes ensuring that temperature of sand bath remained between 180°C-200°C. After 30 minutes, solution was cooled to room temperature. Acetone (5mL) was added and the solution was stirred for 10 minutes. Acetone (10mL) was added once again and stirred until solid dissolved. Reaction progress was monitored by TLC (80% ethyl acetate/20% hexanes) and after stirring was complete the solvent was evaporated using a stream of nitrogen. Residue was purified via column chromatography (100% ethyl acetate). Twelve 10mL fractions were collected and monitored by TLC (1-8 100% ethyl acetate, 9-12 50% ethyl acetate/50% hexanes). Fractions (1-8) that contained the fluorescein product were combined. Solvent was evaporated to yield brownish orange powder solid (.170g, 77.3%); (mp: 310°C-313°C ); 1H NMR (60 MHz, DMSO) δ 6.2 (s, 4H), 6.5 (s, 2H), 7.7 (d, 1H), 7.8 (m, 1H), 7.9 (m, 1H), 8 (d, 1H); 1H NMR (400 MHz, DMSO) δ 6.6 (s, 4H), 6.7 (s, 2H), 7.3 (d, 1H), 7.7 (t, 1H), 7.8 (t, 1H), 8 (d, 1H), 10.2 (s, 2H); UV/Vis λmax 498.6
The purpose of this experiment was to determine the relationship between tail spine length and hemoglobin levels as well as the relationship between tail spine length and heart rate. The concentration of the hemoglobin in Daphnia is dependent on the oxygen available to them.
The SN1 mechanism leads to substitution products, and the E1 mechanism leads to formation of alkenes, therefore in this case, it is shown that this mechanism leads to a substitution of products since the Cl- ion is replacing the OH group by the addition of a strong acid (HCl). When the nucleophile
Green fluorescent protein (GFP) comes from the jellyfish Aequorea Victoria is rare proteins with high fluoresce and absorbance. The purpose this experiments is to purify and express a His2-tagged recombinant from of GFP (rGFP) from the E. coli strain BL21(DE3)< pRSETA-GFPUV > through a series of experiments by using Ni+2 agarose affinity chromatography technology. The GFPuv gene (UV-optimized GFP) was over expressed in the E. Coil strain BL21 (DE3) (pLysS) as an n-terminal His6/Xpress epitope tagged bind protein. Then using Ni2+ Agarose affinity chromatography to obtain purification of the crude extract. Then observe under the long wavelength UV light, the activity of the rGFP in the column fraction. Bradford assay was performed to obtain the total protein amount. When calculating the
Every cell transports materials in and out throught something called a membrane. There are many different methods of transport in the cell Saccharomyces cerevisiae (Serrano, 1977) We want to know does adding higher concentrations of azide more effectively block dye transport? We tested the transport of dye in yeast cells with a metabolic inhibitor. When we did this we showed no difference in the absorbance between different azide solutions, and our control. From this we concluded that azide has no effect on the transport through a yeast cell membrane.
In the Dyes and Crimes laboratory experiment, the phosphorescence, fluorescence, and chemiluminescence properties i.e. traits of several chemicals were examined using (UV) Ultraviolet lamp. Observations on color, intensity, duration of glow, etc. were analyzed to determine the traits of the several chemicals. Correspondingly, the author of the unsigned note was determined through ink extraction, (TLC) Thin Layer Chromatography, (UV-Vis) Ultraviolet-visible spectroscopy along with the Rf values for of each individual sample of ink compared to the unknown: the ink of the author on the unsigned note. Phosphorescence substances
The experiments conducted for this lab report focused on water contamination and filtration. Experiment 1 was effects of groundwater contamination. Oil, vinegar, and laundry detergent were added to clean water with no means of filtration. The clean water was found to be contaminated. A filtration system consisting of cheesecloth and 60 ml of soil was created and the contaminated samples were filtered through it. The soil and cheese cloth did not affectively filter the contaminants. Experiment 2 focused on
The Uptake of Neutral red dye by saccharomyces cerevisiae in the presence of a metabolic inhibitor
When forming the Grignard reagent, the solution became brown in color and as it proceeded the solution became lighter. After a week, the solution was pink/red with clear/white crystals. In the addition of HCl, the solution began to become yellow and two layers began to form. The top layer was yellow and the lower layer was clear/white. The final alcohol product yield was calculated to be 88.317 percent by dividing the weight of the product (6.789 g) by the theoretical yield of 7.69 g. When placed in the infrared spectroscopy machine, alcohol (υO-H) was found at wavenumber of 3366.75 cm-1
b.Describe how the organism would look if it is stained with nigrosin. How does this stain interact with the cell?
In Table 1, fluorene travels further up the TLC plate versus fluorenone since a more nonpolar 80:20 pentane-ether developing solvent is used. Since fluorine is more nonpolar than 9-fluorenone, it will be more soluble in the highly nonpolar solvent and enter the mobile phase easier than the polar 9-fluorenone. Fraction 2 is expected to have two different spots because this is the collection of the eluted solution between Fraction 1 and Fraction 3, where the fraction contains both the remaining fluorene and 9-fluorenone. Fraction 1, however, should not contain any 9-fluorenone since if correctly performed only fluorene and pentane should have eluted out in the mobile phase. A crack in the column during the elution of Fraction caused one section
Chemiluminescence is a reaction well known to those who enjoy crime scene shows. Luminol is first synthesized in the experiment. Then, the crude luminol produced is oxidized to release photons giving off a light blue glow in a dark room. In a crime scene show, the blood acts as a catalyst aiding in the oxidation of luminol and hydrogen peroxide resulting in the blue glow 1.
The name given to E.coli was Bacilli because it is a rod- shaped bacteria. Cocci was given to S. aureus because its structure appears spherical.
Garner, C., George, C., & Primrose, P. (2017). CHE 3238 Organic Chemistry Laboratory Manual – Baylor University (Fall 2017, Spring and Summer 2018 ed.).
This experiment demonstrated the separation of pigments based on relative polarity and proved to be a substantial way to separate compounds. The results were much like that of an experiment performed, which separated carbohydrates in a very similar method with the use of paper chromatography (Inome, Y., & Yamamoto, A.). Proper pipetting technique, which is described by John Husler, was also demonstrated in this experiment. The technique was followed as to prevent contamination and deliver the right amount of solution each time (John Husler: 1983).
Data were collected in September and November 2017 at the Elk Valley Preserve and Field Station in Banner Elk, NC. The preserve is located at 36°10'17.2"N 81°54'45.9"W, in Avery County in western North Carolina. The area is a mixed deciduous forest at an altitude of 1,127 m above sea level. Data were collected in early fall, during which temperatures average around 10-18 degrees Celsius. Summers in Banner Elk are typically mild, with temperatures averaging 21 degrees Celsius, and winters are cold and snowy, averaging near zero degrees Celsius. During the months of September and November there is an overall average precipitation of 12-25cm.