...from cells through different techniques and their presence can be tested through different qualitative tests. The sample choice is egg yolk and was used as a source of lipids in the experiment. Liquid-liquid extraction, separation of the organic and aqueous layer was used to extract the supernatant or extract. Also, thin layer chromatography or TLC was used to separate the different lipid components by using the Rf values computed. The farther the distance traveled by the compound (higher Rf), the more nonpolar the component, while the smaller the distance traveled, the more polar the component (lower Rf). Lecithin and cholesterol was not able to travel the plate. After, the isolated lipid was subjected to qualitative tests such as Acrolein test, test for phosphates, Leibermann-Burchard test and test for unsaturation. Acrolein tests determine the presence of glycerin; the test for phosphate detects phosphate groups in the structure of the lipid, Leibermann-Burchard uses cholesterol as the standard and detects the presence of steroids and terpenes. The test for unsaturation on the other hand detects the presence of double bonds through halogenation reaction. The extract contains glycerol, phosphate, cholesterol and double bonds. Errors in the experiment can be attributed to the human error, false methodology and presence of contaminant in the reagent used....
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...determine which two amino acids where contained within an unknown mixture by comparing the results of a primary amine test, an amide test, a benzene ring test, a thiol test, and paper chromatography to that of amino acids with known compositions. I hypothesize that alanine and lysine will test positive for primary amines because they are the only amino acids being tested in this reaction that contain a primary amine (-NH2). I hypothesize that glutamine will test positive for amides because it is the only amino acid being tested in this reaction that has an amide, or (R1-N-R2). I hypothesize that tryptophan will test positive for benzene rings because it is the only amino acid being tested that contains a benzene ring. I hypothesize that cysteine will test positive for thiols because it is the only amino acid being tested with a thiol, or (R-S-H). I also hypothesize that the hydrophilic amino acids will have smaller Rf values than the hydrophobic amino acids because the chromatography paper is more hydrophilic than the solvent being used, so they will have a higher affinity for the paper and will not travel very far with the solvent. It is important to be able to identify unknown amino acids so that it can be determined which amino acids are found in cells and proteins. Methods: Paper Chromatography 1. Pick either unknown A, B, or C. 2. Draw a pencil line along a long side 2cm from the edge and dot 5uL of the following samples on the line 2cm apart. a. Unknown ...
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...Isolation and Characterization of Proteins: Hydrolysis and Chromatography Niña Maria Inez Torno, Kimberly Victoriano, Steven John Villafuerte, Klarissa Viloria, Liana Yamzon Group 8 2F-Pharmacy Biochemistry Laboratory ABSTRACT Chemical analysis can be either qualitative or quantitative in nature. In qualitative analysis, we work to identify the substances present in a given sample. We are only concerned whether certain substances are present or absent while in quantitative analysis, we are concerned in determining the amount of each component present in a sample. This experiment aims to isolate and characterize the intact protein in wheat flour which is Gluten through hydrolysis and chromatography. Gluten was isolated then hydrolyzed by proteolytic enzyme and was subjected to different qualitative tests. Paper Chromatography was also performed to analyze the different amino acid components of gluten. Tests showed that Gluten has several amino acid components. INTRODUCTION Proteins are a class of organic compounds which are present in and vital to every living cell. In the form of skin, hair, callus, cartilage, muscles, tendons and ligaments, proteins hold together, protect, and provide structure to the body of a multi-celled organism. In the form of enzymes, hormones, antibodies, and globulins, they catalyze, regulate, and protect the body chemistry. In the form of hemoglobin, myoglobin and various lipoproteins, they affect the transport of oxygen and other substances...
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...Cell Processes: Photosynthesis Part 1: Pigments in Plant Leaves Purpose: The purpose of this experiment is to visualize the pigments present in plant leaves by using paper chromatography. Materials Used: Chromatography vial with cap, wax pencil, disposable transfer pipet, chromatography solvent, chromatography paper strip, sharp pencil, ruler, pair of scissors, piece of fresh pre-soaked spinach, coin, pair of forceps. Observations and Data: Distance of Pigments From Original Pencil Line | | Pigment | Color | Migration Distance (mm) | Rf Value | | Carotene | Orange | 59 mm | .94 | | Xanthophyll | Yellow | 56 mm | .89 | | Chlorophyll a | Light Green | 29 mm | .46 | | Chlorophyll b | Dark Green | 14 mm | .22 | Solvent | | 63 mm | | Analysis and Results: 1. Which pigment migrated the farthest on the chromatogram? Explain how this migration occurred. According to the data Carotene travel furthest, this is because it was the most soluble and it didn't form bonds with the paper. 2. What does the Rf value represent? If you were to perform your experiment on a chromatography paper twice the length of the one used, would your Rf values still be the same? Rf value represent the distance traveled by the pigments divided by distance traveled by solvent. The Rf value would stay the same because if the pigments travel further so will the solvent. 3. How do plant pigments and the absorption spectrum relate to photosynthesis? In a plant there are...
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...AccuData® GTS/GTS The Advantage test strip contains the enzyme glucose dehydrogenase which converts the glucose in a blood sample to gluconolactone. This reaction liberates an electron that reacts with a mediator. The oxidized form of the mediator hexacyanoferrate (III) accepts the electron, forming the reduced form of the mediator, hexacyanoferrate (II). The Advantage test strip employs the electrochemical principle of biamperometry. The monitor applies a voltage between two identical electrodes, which causes the reduced mediator formed during the incubation period to be reconverted to an oxidized mediator. This generates a small current that is read by the monitor. If controls do not fall within the acceptable range after one repeat 1 Do not report the patient results (this would be if the operatoris running controls for the purpose of checking the meter; otherwise, the QC lockout would not have allowed the patient to be tested prior to getting the controls in acceptable range). 2 Review the test policy and procedure. 3 To get a trade, take all of the reagents and supplies out of the drawer and take the entire GTS to the High Volume Lab of Chemistry, 5th floor McCullough Bldg. 4 If feasible for the testing site, the Nurse Manager/designee or Clinic Manager can authorize the borrowing of a GTS unit from another location. cont. NOTE: The proper comment code (09) identifying a temporary GTS unit is being used should be entered after the patient result is displayed...
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...Name: Srikant Tulsi Reg # 10/0705/0497 Experiment # 4 Date: 5/11/2014 Partners: Shefali Seecharran Pride Ade-Thomas Claude Fraser Title: Synthesis of 9,10-Dihydroanthracene-9,10-Endo-α,β – Succinic Anhydride Reactions: Reaction Mechanism Physical Properties Table: Chemical | Formula | M.W (g/mol) | Quantity (g or ml) | Amount( moles) | Molar Equivalent | M.P(OC) | B.P (OC) | Density\(g/cm3) | Yield (g) | % yield | Anthracene | C14H10 | 178.23 | 2.018 g | 0.0113 | 1 | 215 | 339 | 1.28 | | | Xylene | C8H10 | 106.16 | 2.5 ml | 0.0022 | 1 | -47.4 | 138.5 | 0.864 | | | Maleic Anhydride | C4H2 O3 | 98.06 | 1.001 g | 0.0102 | 1 | 52.8 | 202 | 1.480 | | | Hexane | C6H14 | 86.18 | - | - | | -96 | 68 | 0.655 | | | DCM | C10Cl2 | 84.93 | - | - | | -96.7 | 39.6 | 1.327 | | | Ethyl Acetate | C4H8O2 | 88.11 | - | - | | -83.6 | 77 | 0.897 | | | 9,10-Dihydroanthracene-9,10-Endo-α,β – Succinic Anhydride | C18H12O3 | 276.29 | - | - | | 262-264 | | | 2.326 | 82.43 | Calculations: * Calculating the number of moles of Anthracene Mass of Anthracene Used is 2.018 g Molar Mass is 178.23 g/mol Number of moles = MassMolar mass = 2.018 g 178.23gmol = 0.0113 moles * Calculating the number...
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...Name: Srikant Tulsi Reg # 10/0705/0497 Experiment # 4 Date: 5/11/2014 Partners: Shefali Seecharran Pride Ade-Thomas Claude Fraser Title: Synthesis of 9,10-Dihydroanthracene-9,10-Endo-α,β – Succinic Anhydride Reactions: Reaction Mechanism Physical Properties Table: Chemical | Formula | M.W (g/mol) | Quantity (g or ml) | Amount( moles) | Molar Equivalent | M.P(OC) | B.P (OC) | Density\(g/cm3) | Yield (g) | % yield | Anthracene | C14H10 | 178.23 | 2.018 g | 0.0113 | 1 | 215 | 339 | 1.28 | | | Xylene | C8H10 | 106.16 | 2.5 ml | 0.0022 | 1 | -47.4 | 138.5 | 0.864 | | | Maleic Anhydride | C4H2 O3 | 98.06 | 1.001 g | 0.0102 | 1 | 52.8 | 202 | 1.480 | | | Hexane | C6H14 | 86.18 | - | - | | -96 | 68 | 0.655 | | | DCM | C10Cl2 | 84.93 | - | - | | -96.7 | 39.6 | 1.327 | | | Ethyl Acetate | C4H8O2 | 88.11 | - | - | | -83.6 | 77 | 0.897 | | | 9,10-Dihydroanthracene-9,10-Endo-α,β – Succinic Anhydride | C18H12O3 | 276.29 | - | - | | 262-264 | | | 2.326 | 82.43 | Calculations: * Calculating the number of moles of Anthracene Mass of Anthracene Used is 2.018 g Molar Mass is 178.23 g/mol Number of moles = MassMolar mass = 2.018 g 178.23gmol = 0.0113 moles * Calculating the number...
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...AP® Investigation #5 Cell Processes: Photosynthesis –STUDENT GUIDE Kit # 36W7405 Table of Contents Background. . . . . . . . . . . . . . . . . . . . . . . . . . 1 SAFETY PRECAUTIONS. . . . . . . . . . . . . . . . . . . . . . 3 Part 1: plant pigments and chromatography (Structured inquiry). . . . . . . . . . . . . . . . . . . . . 4 Part 2: floating disc assay (Guided Inquiry) . . . . . . . . . 6 Part 3: design an experiment (OPEN inQUIRY) . . . . . . . . . 9 MATERIAL SAFETY DATA SHEETS . . . . . . . . . . . . . . . . . 11 **AP® and the Advanced Placement Program are registered trademarks of the College Entrance Examination Board. The activity and materials in this kit were developed and prepared by Ward’s Natural Science Establishment, which bears sole responsibility for their contents. Copymaster. Permission granted to make unlimited copies for use in any one school building. For educational use only. Not for commercial use or resale. US: www.wardsci.com Canada: www.wardsci.ca ©2012, Ward’s Natural Science All Rights Reserved 250-7455 v.7/12 AP® Investigation #5: Cell Processes: Photosynthesis –STUDENT GUIDE Kit # 36W7405 Background OBJEcTIVES ‹ Design a plan for collecting data to show that all biological systems are affected by complex biotic and abiotic interactions. ‹ Use models to predict and justify that changes in the subcomponents of a biological polymer affect...
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...In biology, in order to purify biological molecules, a common process called chromatography is often used. This process involves the separation of the large particles in the solution from the smaller particles. This is instrumental in filtering the solution, but without specialty scientific instruments, there is no way to immediately know the concentration of the, now separated, solution. However, there are multiple processes that are utilized to find unknown concentrations of a solution. One of these, known as the Bradford method, allows for an accurate, fairly simple, and timely assessment of these unknown concentrations by using a dye called the Coomassie Brilliant Blue G-250. The samples that contain this dye are run through a spectrophotometer,...
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...Pigment Color | Pigment Type | Distance Traveled by Pigment / a (cm) | Distance Traveled by Solvent / b (cm) | Rf Value ab | Yellow | Carotene | 6.2 | | | Yellow-brown | Xanythophyll | 4.0 | | | Blue-green | Chloropyhll a | 2.9 | | | Green | Chloropyhll b | 1.3 | | | RESULT: Table 1: The result of the experiment is to determine the colour of the pigment on filter paper by paperchromatography. Besides, the a and b which represents the distance travelled by pigment and distance travelled by solvent were measured and recorded in the table. The Rf value was then calculated by using the formula ab. A= distance moved by the pigment from its original position B= distance moved by solvent from the same position. *Round Rf values to 2 decimal places CONCLUSION: We used paper chromatography to separate the pigments found in spinach. Since plants use several different pigments to capture light energy for the light dependent reaction in photosynthesis, we expected that we would see several different bands of pigments on our chromatography paper. As the data in Table 1 indicates, we obtained four bands of different pigments from the spinach leaf: yellow- brown, light green, green, and yellow. The bands moved different distances up the chromatography paper, and were of different widths. According to the data, our hypothesis was correct. Different types of pigments are present in plants. The distances traveled by the pigments can indicate how big...
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...quantities of nicotine as small as 0.4 microgram. INTRODUCTION During the course of our study on the chemical patterns of plant growth and development (Peters et al., 1972), it was necessary to use rapid procedures to analyze chemical contents of growing tissues. The present study deals with the development of a procedure for the rapid detection of the alkaloids. The histochemical detection of alkaloids in growing tissues, as demonstrated by others (Chaze, 1932; James, 1950), is based on the reaction with iodine in potassium iodide solution. These methods are complicated by the presence of carbohydrates and proteins, which also react positively to iodine in potassium iodide solution. James (1946) was able to overcome the difficulty of liberating alkaloids from denatured proteins by blotting the tissue on filter paper prior to other treatment; however, the blotting procedure results in partial loss of cellular alkaloid content. A tedious solvent extraction procedure followed by thin-layer chromatography has been used by Speake et al. (1964). Other workers have used the reliable but time-consuming steam-distillation extraction technique (Solt, 1957; Brown and Byerrum, 1952). This paper describes an improved procedure whereby tobacco...
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...transmittance by/through plant chlorophyll and accessory pigments. Photosynthesis is the process by which plants use energy from light (i.e. the sun, photons, electromagenetic energy), water, and carbon dioxide to produce ‘food’ for themselves, or some for of sugar, like glucose. Photosynthesis has two stages which take place within the chloroplasts of a plant cell, where the photosynthetic pigments, chlorophyll reside. The first stage occurs in the thylakoid of the chloroplasts and is called ‘the light reactions’. During this stage water is split, the oxygen changes (or is evolved) and energy is harvested through photophosphorylation. Photophosphorylation transforms the light energy into the chemical energy of ATP and NADPH, which aid in the next stage, the Calvin Cycle or light independent reactions. During the Calvin Cycle, carbon dioxide is reduced to glucose and oxygen is released as a by product, at which time the ADP and the NADP+ are returned to the light reactions to start the cycle over again, or regenerate. A simplified equation for photosynthesis is as follows: 6CO2 + 6H20 + Light Energy → C6H12O6 (glucose) + 6O2 Photosynthesis is basically a plants version of cellular respiration and is essential for their survival. During the first stage, light energy or electromagnetic energy/radiation, is absorbed by the chlorophyll, except for the green wavelengths of the visible light spectrum. This is why they appear to be green, or why we see green...
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...to determine if it changed in response to the independent variable. What solutions were used to test for the 4 types of organic molecules? Iodine- Polysaccharide Benedict’s Reagent- Sugar Biuret Test- Protein Brown Paper Test- Lipids Vegetable Oil- Solubility of Lipids What does a positive test look like? -Iodine test for polysaccharide: dark purple/black/blue -Vegetable Oil test for solubility of lipids: 1 layer -Biuret test for protein: violet color -Benedict’s Reagent for sugar: very high concentration/orange-red How do you convert Celsius to Fahrenheit and vise versa? Degrees Fahrenheit= 9/5 degreesC + 32 degrees Degrees Celsius= 5/9(degreesF - 32 degrees) What is the compound scope magnification equation? eyepiece mag x objective mag What is the resolution equation?...
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...composition, structure and function. However, they share a common feature as each macromolecule is composed of repeating subunits, characteristic of the macromolecule. The subunits are linked together by a bond between two adjacent subunits, formed by the loss of water (condensation). Thus, macromolecules can be broken down by the addition of water across the bond, in a process known as hydrolysis. This process was used in the experimental procedure to allow analysis of each individual macromolecule in its subunit form. Proteins are hydrolyzed into amino acids, nucleic acids are hydrolyzed into sugar, base and phosphate, and polysaccharides are broken down into simple sugars. In the second part of the experiment, the principle method of chromatography was used to analyze the macromolecules isolated in part one of the experiment. With this technique, individual molecular species were separated from one another. This separation technique was performed for the protein and nucleic acid components. In addition, for each case, a series of knowns including one unknown...
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...obtain information about the protein. The myoglobin extracted was brownish red in color. The isolated protein was subjected to autoclaving for five hours at 15 psi. After autoclaving, the resultant solution was clear and colorless. The acid component (HCl) added prior to autoclaving was neutralized by the addition of a base, 1-molar NaOH. The neutralized acid hydrolysate was used for characterization tests and chromatography. INTRODUCTION Proteins, from the Greek proteios, meaning first, are a class of organic compounds which are present in and vital to every living cell. In the form of enzymes, hormones, antibodies, and globulins, they catalyze, regulate, and protect the body chemistry. In the form of hemoglobin, myoglobin and various lipoproteins, they effect the transport of oxygen and other substances within an organism.(1) Myoglobin is an extremely compact heme protein found primarily in cardiac and red skeletal muscles. It functions in the storage of oxygen and facilitates the transport of oxygen to the mitochondria for oxidative phosphorylation. (2) Hydrolysis is performed to obtain information about the said protein. (3) It is a process in which the compound reacts with water leading to its decomposition. It takes place via Nucleophilic Acyl Substitution. Nucleophilic Acyl Substitution involves the attack of nucleophiles to acid carbonyls of the compound. This reaction would yield substitution products. EXPERIMENTAL A. Sample/s used: The sample...
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