...from spinach leaves (including chlorophyll a, chlorophyll b and beta carotene). The experiment also compared and contrasted two different methods of separation. The first part of the experiment explored column chromatography and the second part explored thin-layer chromatography. Introduction: (3 marks)* The three pigments that can be extracted from spinach (chlorophyll a, chlorophyll b and beta carotene) all differ based on their polarity, which arises from functional groups in their structure. The most effective way to separate pigments from spinach is based on their differences in their polarities and this lab explored two chromatographic methods to do so. Chromatography is the technique of separating organic mixtures by allowing them...
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...The purpose of this lab was to purify crude product by column chromatography. The crude product to be purified in this lab was acetylferrocene contaminated with ferrocene. Column chromatography is a technique that utilizes the liquid solid absorption method. This technique has the advantage by isolating more of the product that need to be analyzed. In Column Chromatography the mixture that is being examined, mixture of our product or compound is dissolved in small amount solvent that is then placed on top of the column. Finely packed solid absorbent (silica gel) act as the stationary phase. In order for the mixture to move down the column a eluting solvent (mobile phase) is placed. The separation of the organic compounds depends on how...
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...Jacobsen that the chiral Jacobsen’s Salen(Co) complex 6 is effective in acting as a catalyst for the hydrolytic kinetic resolution (HKR) for regioselecting epoxides. The complex is prepared in four steps. A mixture of 1,2-diaminocyclohexane isomers was selectively crystallized to form 1,2-diaminocyclohexane L-tartrate salt. The salene ligand was formed by reacting 3,5-di-tert-butyl-2-hydroxybenzaldehyde with 1,2-diaminocyclohexane. The Salen(Co) complex was formed by treating the salen ligand with cobalt(II) acetate. Finally, the catalyst was formed by processing the ligand through aerobic oxidation in the presence of acetic acid. The catalyst was used to resolve phenyl glycidyl ether and the diol 9 and epoxide 8 were separated by flash chromatography. The products were characterized through chiral HPLC, IR, H-NMR, and polarimitry tests. HPLC spectrum showed that the catalyst had successfully yielded enantiomerically pure diol 9 and epoxide 8 molecules with 8 being resolved to 99.4% ee and 9 with 93.8% ee. Although (R,R) and (S,S) Jacobsen’s Salen(Co) Catalysts select for different enantiomers of epoxides, reactions utilizing either catalyst resulted in similar yields, within 1%. The data collected is useful in showing that the HKR reaction was highly effective in regioselecting the respective epoxides. Introduction Despite their opposite structures, enantiomers posses identical chemical properties and only differ by their physiological activity in chiral environments, one being...
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...Types of Chromatography Adsorption Chromatography Adsorption chromatography is probably one of the oldest types of chromatography around. It utilizes a mobile liquid or gaseous phase that is adsorbed onto the surface of a stationary solid phase. The equilibriation between the mobile and stationary phase accounts for the separation of different solutes. Partition Chromatography This form of chromatography is based on a thin film formed on the surface of a solid support by a liquid stationary phase. Solute equilibriates between the mobile phase and the stationary liquid. Ion Exchange Chromatography In this type of chromatography, the use of a resin (the stationary solid phase) is used to covalently attach anions or cations onto it. Solute ions of the opposite charge in the mobile liquid phase are attracted to the resin by electrostatic forces. Molecular Exclusion Chromatography Also known as gel permeation or gel filtration, this type of chromatography lacks an attractive interaction between the stationary phase and solute. The liquid or gaseous phase passes through a porous gel which separates the molecules according to its size. The pores are normally small and exclude the larger solute molecules, but allows smaller molecules to enter the gel, causing them to flow through a larger volume. This causes the larger molecules to pass through the column at a faster rate than the smaller ones. Affinity Chromatography This is the most selective type of chromatography employed...
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...Separation techniques Chromatography Chromatography is a separation technique used to separate the different components in a liquid mixture. It was introduced by a Russian Scientist Michael Tswett. Chromatography involves the sample being dissolved in a particular solvent called mobile phase. The mobile phase may be a gas or liquid. The mobile phase is then passed through another phase called stationary phase. The stationary phase may be a solid packed in a glass plate or a piece of chromatography paper. The various components of the mixture travel at different speeds, causing them to separate. There are different types of chromatographic techniques such as column chromatography, TLC, paper chromatography, and gas chromatography. Paper chromatography is one of the important chromatographic methods. Paper chromatography uses paper as the stationary phase and a liquid solvent as the mobile phase. In paper chromatography, the sample is placed on a spot on the paper and the paper is carefully dipped into a solvent. The solvent rises up the paper due to capillary action and the components of the mixture rise up at different rates and thus are separated from one another. Distillation Simple distillation is a method used for the separation of components of a mixture containing two miscible liquids that boil without decomposition and have sufficient difference in their boiling points. The distillation process involves heating a liquid to its boiling points, and transferring...
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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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...Torres 1 Luis A. Torres Group #11 USC Chemistry 322b Formal Lab Report 6th November 2015 I. II. Enzymatic Resolution of 1-Phenylethanol and Diastereomer Analysis Objective/Abstract Enzymatic transesterification reaction was performed to study the resolution of diastereomers using 1H-NMR analysis. The stereo-selectivity of acylase I, an enzyme, for a 50:50 racemic mixture of 1-phenylethanol was determined. In the first of a two-step reaction, 1-phenylethanol was reacted with vinyl acetate with the help of acylase I to form an ester, unreacted 1-phenylethanol, and vinyl alcohol. The unreacted 1phenylethanol was separated from the ester by column chromatography and confirmed by thin-layer chromatography (TLC). In the second reaction, the unreacted 1-phenylethanol was reacted with (R)-(-)-acetoxyphenylacetic acid to form a diastereomer ester. In the latter reaction, four different 1-phenylethanol samples were used in order to compare 1HNMR data of the resulting diastereomer esters and determine which enantiomer of the 50:50 racemic mixture was preferred by acylase I. Those four samples were: (1) racemic 1-phenylethanol, (2) unreacted 1-phenylethanol, (3) (R)-1-phenylethanol, and (4) (S)-1phenylethanol. After 1H-NMR analysis, it was found that the (S)-1-phenylethanol was preferred by acylase I. III. Introduction and Background Information Scheme 1: Reaction of 1-phenylethanol with vinyl acetate in the presence of the enzyme acylase I to produce 1-phenylethyl...
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...Paper and Column Chromatography Report Guidelines 1. (1) – Distance from origin compound (2) –Distance from origin to solvent front (3) –Retention factors of known an unknown (4) –polarity of side chain (5)-relative size | Tyro | Ser | Leu | Lys | Tryp | Asp | Glyc | Ala | unknown | (1) | 4.74 cm | 3.93 cm | 6.67 cm | 3.34 cm | 5.85 cm | 3.32 cm | 3.86 cm | 4.12 cm | 3.79 cm | (2) | 8.72 cm | 8.88 cm | 8.72 cm | 8.71 cm | 8.76 cm | 8.54 cm | 8.71 cm | 8.70 cm | 8.87 cm | (3) | 0.544 cm | 0.443 cm | 0.765 cm | 0.383 cm | 0.668 cm | 0.384 cm | 0.443 cm | 0.474 cm | 0.427 cm | (4) | polar | polar | Non polar | Mediumpolar | polar | polar | Nonpolar | Non polar | Medium polar | (5) | 13 | 7 | 9 | 10 | 17 | 9 | 5 | 6 | 10 | 2. Code: CHI, The conclusion of the unknown substance is that it is Lysine. The reasoning is based primarily off of the paper chromatography experiment. 3. In paper chromatography, the stationary phase is a very uniform absorbent paper. The mobile phase is a suitable liquid solvent or mixture of solvents. But, the stationary phase is more polar because the paper is not actually polar at all, compared to, probably, water as the mobile phase. 4. 5. 6. Tryptophan, Tyrosine, and Lysine. Listed from largest to smallest they all are polar, with the smallest, Lysine, having a medium polarity. 7. 8. 9. references: http://www.macalester.edu/~kuwata/Classes/2001-02/Chem%2011/Revised%20Amino%20Acids%20(9%201%2001).pdf http://www...
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...http://www.labcompliance.com/tutorial/methods/default.aspx?sm=d_d TutorialHome | Contact Us | Newsletter | Usersclub | Books | Audio Seminars Seminars Audio Seminars Video Seminars Workshops Literature Books SOPs Validation Examples Free Literature Glossary Usersclub Intro Log-in Register Preview Renewal Tutorials Risk Management Practices Computer Validation Part11 Method Validation ISO 17025 Lab Equipment Qualification Good Laboratory Practices About About Labcompliance Contact Labcompliance Scope Tax/Bank Information All come with 10+ Best Practice Documents: SOPs, Checklists, Examples Transfer of Analytical Procedures According to the New USP Chapter <1224> With SOPs, templates and examples for easy implementation March 21, 2013 Quality by Design (QbD) for Analytical Method Development and Validation Learn how to design robustness for easy transfer and to avoid OOS situations Recorded, available at any time Validation of Analytical Methods for GLP and Clinical Studies Learn how to design, prepare, conduct and document for FDA compliance Recorded, available at any time Eight Steps for Cost-effective Laboratory Compliance Up-to-date overview, hot topics and trends. Recorded, available at any time Verification of Compendial Methods according to the New USP Chapter <1226> Understand the new risk based approach and and get real world case studies for testing Recorded, available...
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...Kerry Nguyen Experiment 9: Preparation of Acylferrocene, by Friedel-Crafts Acylation and Purification by Column Chromatography Balanced Equation for the Reaction Carried Out Using Structural Formula: Name of Compound Prepared: YIELD DATA: Compound | Theoretical Yield | Actual Yield | Percent Yield | Ferrocene | 0.094 | 0.043 | 45.74% | Acetylferrocene | 0.115 | 0.017 | 15.65% | Theoretical Yield: Mass of Ferrocene weighed/used: 0.094g Mass of Acetylferrocene calculated 0.094gferrocene11moleferrocene186.04gferrocene1moleacetylferrocene1moleferrocene228.08gacetylferrocene1moleacetylferrocene=0.11524gacetylferrocene Actual Yield: Ferrocene : 26.699g – 26.656g = 0.043g Acetylferrocene: 37.811 – 37.794 = 0.017g Percent Yield: Ferrocene 0.043g0.094g∙100%=45.74% Acetylferrocene 0.017g0.115g∙100%=15.65% PHYSICAL PROPERTIES: Powder/grainy, dark brown-orange Literature melting point: 172.5 oC Literature boiling point: 249 oC Powder/grainy, orange-yellow Literature melting point: 85 – 86 oC Literature boiling point: 161 oC ANALYSIS OF SPECTRA AND CHROMATOGRAMS OF YOUR PRODUCTS: Solvent (mixture) used to develop plate – 3:1 toluene:EtOH Compound spotted | Spot 1: Rf | Spot 2: Rf | Spot 3: Rf | Ferrocene | 7.55cm/8.20cm=0.92 | 0.92 | 0.92 | Acetylferrocene | 3.17cm/8.20cm=0.39 | 0.39 | 0.39 | Compound | Melting Point (lit. M.P.) | Ferrocene | 164 – 171.5 oC (lit. 172 – 174 oC) | Actylferrocene | 78.9 - 81...
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... . . . . . . . . . . . . . . 2.4.2 Diet . . . . . . . . . . . . . . . . . . . . . . . 2.4.3 Using Drugs to Reduce Detection Times . . . 3 Test Methods 3.1 Substances that are Detectable . . . . . . . . . . . 3.2 DrugAlert . . . . . . . . . . . . . . . . . . . . . . . 3.3 Gas Chromatography . . . . . . . . . . . . . . . . . 3.4 Gas Chromatography / Mass Spectrometry . . . . . 3.5 Hair testing . . . . . . . . . . . . . . . . . . . . . . 3.6 High Performance Liquid Chromatography . . . . . 3.7 ImmunoAssay . . . . . . . . . . . . . . . . . . . . . 3.7.1 Radio ImmunoAssay (aka AbuScreen) . . . 3.7.2 Enzyme Multiplied Immunoassay Technique 3.7.3 Fluorescence Polarization ImmunoAssay . . 3.8 PharmChek . . . . . . . . . . . . . . . . . . . . . . 3.9 TestCup . . . . . . . . . . . . . . . . . . . . . . . . 3 11 13 13 13 15 16 16 16 17 17 18 18 18 19 19 21 21 21 22 24 24 25 25 25 26 26 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 CONTENTS 3.10 Thin Layer Chromatography . . . . . . . . . . . . . . . . . . . 26 4 Test...
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...CHROMATOGRAPHY Chromatography (from Greek word Chroma means “color” and Graphein means “to write”) is the collective term used for the separation of the mixture. The mixture is dissolved in a fluid called the mobile phase, which carries it to another material known as the stationary phase. The separation is based on the speed of different constituents of the mixture and on the differential partition of the mobile and the stationary phase (web link7). Distribution or partition coefficient is the basis of chromatography which describes a way in which constituents of a mixture distributes between two immiscible phases. For two such phases A and B, the value for this coefficient is a constant at a given temperature and is given by the expression (36): Kd = Concentration in phase A Concentration in phase B COLUMN CHROMATOGRAPHY For separations of solid and liquids, column chromatography is one of the most important techniques. It is a solid liquid separation technique in which the stationary phase is solid and the mobile phase is liquid. It is based on the principle of differential adsorption of the substance by adsorbent (web link 8). The stationary phase is packed into a glass or metal column and mixture of analytes are then applied, the mobile phase (eluent) is passed through the column either by use of a pumping system or applied gas pressure. The stationary phase is either coated onto discrete small particles (the matrix)...
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...CLC204 – Analytic Chemistry Experiment Quantitative Analysis of an Unknown Liquid Sample Objective: To be familiar with Ultra Violet Spectrophotometer (UV) and Gas Chromatography (GC) used in chemical analysis UV Abstract In this report, people who consumed tom yum soup suffered from nausea and vomiting due to copper poisoning which was found to leach from a pot into the soup under high heat and acidic condition. It was also suspected that an organic compound was present in the soup which enhanced the absorption of copper in consumer. Hence, as an analytical chemist, we have to use UV to determine the actual concentration of copper standards and blank using external calibration standards. The result of the test solution was measured by comparing it with the calibration of copper. Introduction The goal of this experiment is to obtain the concentration of copper in a known solution. Therefore, in this experiment, we will be using UV to measure the absorbance of the solution. A spectrophotometer is used to measure the amount of light that a sample absorbs. Ultraviolet (UV) light is an electromagnetic radiation with a wavelength in the range of 10nm to 400nm and energy from 3eV to 124Ev. With a shorter wavelength as compared to visible light, UV light is able to penetrate more readily through obstacles. Its name came from a spectrum which humans identify as the colour violet. UV light is invisible, but it can be seen indirectly when it makes other substances...
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...10 Technology used by Medical Technologist 1. Description: A centrifuge is a piece of equipment that puts an object in rotation around a fixed axis (spins it in a circle), applying a potentially strong force perpendicular to the axis of spin (outward). The centrifuge works using the sedimentation principle, where the centripetal acceleration causes denser substances and particles to move outward in the radial direction. At the same time, objects that are less dense are displaced and move to the center. In a laboratory centrifuge that uses sample tubes, the radial acceleration causes denser particles to settle to the bottom of the tube, while low-density substances rise to the top.[1] There are 3 types of centrifuge designed for different applications. Industrial scale centrifuges are commonly used in manufacturing and waste processing to sediment suspended solids, or to separate immiscible liquids. An example is the cream separator found in dairies. Very high speed centrifuges and ultracentrifuges able to provide very high accelerations can separate fine particles down to the nano-scale, and molecules of different masses. Large centrifuges are used to simulate high gravity or acceleration environments (for example, high-G training for test pilots). Medium-sized centrifuges are used in washing machines and at some swimming pools to wring water out of fabrics. Gas centrifuges are used for isotope separation, such as to enrich nuclear fuelfor fissile isotopes. From: https://en...
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...David O. Johnston David Lipscomb College I II The Methclnolysir of Acetal A chemical kinetics a n d g a r chromatography experiment Nashville, Tennessee The methanolysis of acetal and gas chromatography complement perfectly to provide a rare and interesting experiment for the physical chemistry student. The reaction of acetal with methanol is a consecutive reaction in which diethyl acetal (aretal) replaces its two ethoxy groups with methoxy groups forming methy1et)hylacetal first and dimethyl acetal in the second step. Investigating the reaction kinetics for this type of reaction mas almost impossible before the advent of gas chromatography (I). There are many advantages to using the methanolysis of acetal in this study: Sufficient data for a complete analysis can be collected within three hours. The reaction requires reagents which are readily available in high purity. The reactants, intermediate, and products are liquids at temperatures easily accessible for kinetic runs. The primary reactant, intermediate, and product are sepe rated rleanly on a typical gas chromatographic column. The peak heights are linear with concentration in the range studied (2). Analysis of the data affords an application of firshorder kin~ticsand the steady state hypothesis. The experiment presented herc is an adaptation of the detailed study of the reaction by Juvet and Chiu (2), who have shown it to be acid catalyzed and first order in diethyl acetal and methylethyl acetal. By decreasing...
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