...Chromatography which literally means “color writing” was invented by Mikhail Tsvet, a Russian botanist in 1903 while studying the coloring materials in plant life. It is defined as a laboratory technique utilized by scientists for the separation of organic and inorganic mixtures into their respective components in order to analyze, identify, purify and/or quantify the mixture or components. There are different types of chromatography which are use including liquid chromatography, gas chromatography, ion-exchange chromatography, paper chromatography and thin-layer chromatography. In consonance with these, while each type of chromatography uses different methods to separate compounds, all forms work on the same principle. They all have a stationary...
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...The first of these methods was loss of weight.1 This method, although widely used at the time of its inception, was not the most accurate or effective. This type of analysis was not very involved but had its drawbacks. These included using too much sample, approximately one to two grams, the lack of specifics in the results, and the fact that the atmosphere could alter the results. This is where gas chromatography and micro-solid phase extraction remedied the issue. Although gas chromatography (GS) and solid-phase microextraction (SPME) were not the first types of separation and analytical determinants for identifying and removing residual solvents, they are extremely more accurate and effective compared to the methods that preceded...
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...Chromatography Aim: To test the purity of aspirin using chromatography. Vocational context The results which we have collected while doing this chromatography will be helpful to many different types of people. One group of people who will be interested in the results is doctors when a patient comes to visit the doctor with a virus or something the doctor can look at these results and they will be able to make a decision on how powerful the aspirin to give the patient. Another group of people who might take an interest to these results are drug testers Background science Aspirin also known as acetylsalicylic acid, is a salicylate drug, often used as an analgesic to relieve minor aches and pains, as an antipyretic to reduce fever, and as an anti-inflammatory medication. Aspirin separates and decomposes very quickly in hydroxide and alkali metals. Aspirin is stable in dry air but gradually hydrolyses when it comes into contact with moisture Chromatography can be used to separate mixtures of coloured compounds. Mixtures that are suitable for separation by chromatography include inks, dyes and colouring agents in food. There is two phases the mobile phase which flows through column, carries analyte and the stationary phase which stays in a place, does not move. The separation of the molecules is based on the partitioning between the mobile and stationary phase. Chromatography is used in labs often to separate different compounds A TLC plate is a sheet of glass...
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...separated based on their ligand exchange interactions with these metal ions: due to the very minute differences in energy of the enantiomers (10-14 J mol-1), separation via ligand exchange is possible. Protein selectors, work on the principal that proteins bond to only one enantiomer, as each enantiomer possesses different pharmacological properties. Since a protein is functional only in the presence of a necessary compound, the proteins in a specific CSP will interact only with one type of enantiomer, thus displaying different retention times for each chiral molecule. Brush (Prickle-concept). Come in many varieties, used for different chiral compounds, but all Prickle-type phases work on three principals: aromatic groups, capable of donor-acceptor interaction (acids/bases) polar hydrogen-bonds, and (iii) non-polar groups, for steric repulsion, based on Van der Waal interactions. Though the above CSPs come with great variety for identifying many compounds, this list is not exhaustive, and many more types of CSPs exist; the listed CSPs are just a few of the most common used in chiral...
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...Separating and Determining the Colored Components of Capsicum fructescens Using Column and Thin Layer Chromatography Duran, K., Ednalino, M., Encarnacion, F., Fernandez, T., Fernando, S., Garcia, D. 2D-Pharmacy, Group 4, Department of Pharmacy, Faculty of Pharmacy, University of Santo Tomas, España Boulevard, 1015 Manila, Philippines ABSTRACT Chromatography was used to separate and determine the purity of the colored components present in a substance. Separation of the colored components of Capsicum fructescens (Red Siling Labuyo) using column chromatography, determining the purity of the components using thin layer chromatography (TLC), and measuring the retention factor (Rf) values of the colored components in TLC were the aims of the experiment. Dichloromethane:Hexane (DCM:Hexane) (1:1) was used to extract the pigments of the red siling labuyo. For the column chromatography, 0.5 mL of the extract was placed on top of the column containing silica gel. DCM-Hexane, DCM, and DCMmethanol was introduced in succession. Colored eluates were collected on separate test tubes and noted the number of drops for each. For the thin layer chromatography, the eluates were spotted on a pre-coated TLC plate and was placed on an equilibrated chamber, with DCM:Hexane as the solvent system. After visualizing the results on the TLC plate, the distance traveled by the pigments were computed for the Rf values. The results showed that the yellow pigment had the greatest distance traveled...
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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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...------------------------------------------------- Gas chromatography Gas chromatography (GC), is a common type of chromatography used in analytical chemistry for separating and analyzing compounds that can be vaporized without decomposition. Typical uses of GC include testing the purity of a particular substance, or separating the different components of a mixture (the relative amounts of such components can also be determined). In some situations, GC may help in identifying a compound. In preparative chromatography, GC can be used to prepare pure compounds from a mixture.[1][2] In gas chromatography, the mobile phase (or "moving phase") is a carrier gas, usually an inert gas such as helium or an unreactive gas such as nitrogen. The stationary phase is a microscopic layer of liquid or polymer on an inert solidsupport, inside a piece of glass or metal tubing called a column (a homage to the fractionating column used in distillation). The instrument used to perform gas chromatography is called a gas chromatograph (or "aerograph", "gas separator"). The gaseous compounds being analyzed interact with the walls of the column, which is coated with a stationary phase. This causes each compound to elute at a different time, known as the retention time of the compound. The comparison of retention times is what gives GC its analytical usefulness. Gas chromatography is in principle similar to column chromatography (as well as other forms of chromatography, such as HPLC, TLC), but has several notable...
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...Plan an experiment to investigate if the pigment in red flowers consists of one or more colors. In your investigation think about how you can extract the pigment and check whether it consists of one or more color. PLAN FOR INVESTIGATION A. Purpose of the Investigation The purpose of this investigation is to know whether the pigment in red flowers consist one or more colors. The goal of this experiment is to analyze the pigments found in flowers using paper chromatography. B. Materials/ Apparatus The materials used in this experiment include filter paper, scissors, pencil, ruler, clean jar or drinking glass, large- mouth glass jar, water (at least 60 mL, distilled water is preferable, but tap water is also suitable), 70% isopropyl alcohol, measuring cup, piece of scratch paper, coin, timer or clock, and of course red flower petals. You will need at least 2 flower petals from at least 3 different plants. Tip: Larger petals work better than smaller petals. C. Steps/ Procedure Cut the chromatography filter paper into strips that are about 2 cm wide and as long as the large-mouth glass is tall (the strips should all be the same size). You will want to make at least three strips for each flower you want to investigate, or a minimum total of nine strips. Use a ruler and pencil to draw a line across each paper strip, horizontally, 2 cm from the bottom, as shown in Figure 1. This is the origin line, where the sample will later be spotted. Tip: Do not use a pen for...
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...Introduction: The experiments of Beadle and Tatum proposed a hypothesis that stated that certain gene traits are related to particular enzymes that are missing. Investigation #19 allowed students to investigate and identify the chemical differences within species of fruit flies, in order, to understand the actions of their genes. The following materials were used to visually see the chemical differences in Drosophila melanogaster mutants: the eyes from the flies, paper chromatography, a good solvent for the paper chromatography to interact with and a UV light source. The seven species of flies used include: wild type, white, cinnabar, brown, cinnabar brown double mutant, sepia and white apricot. The phenotypes of these species varied from a red pigment, brown pigment and a white pigment. When a red pigment and brown pigment are crossed, the resulting color is brick red....
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...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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...Chapter 4 Optimisation of a Protein A Chromatography Process Introduction The optimisation of a chromatography, we know, is vital to the efficiency of a separation process. Protein A chromatography by virtue of its incredible ability to achieve high purity of biopharmaceutical products has made itself indispensable for downstream purification of biomolecules (Hammond et al.) Even though many variations of different chromatographic separations have been employed as ‘capture’ steps over the years, Protein A affinity chromatography has come to be used as the method of choice for direct ‘capture’ and purification of monoclonal antibodies. The highly specific binding between the Fc-region of an antibody and Protein A, as elucidated to in Chapter...
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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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...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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... 4. PROCESSES FOR PURIFICATION 1. Crystallization 2. Sublimation 3. Distillation 4. Chromatography 5. Differential Extraction 6. Qualitative Analysis 5. 1. CRYSTALLIZATION Aim To separate a solid compound in pure and geometrical form. Principle A saturated solution of the impure substance in a suitable solvent is made at a temperature higher than the room temperature. On cooling this solution, the substance reappears in the form of well shaped crystals. Process Purification by crystallization involves the following steps: Choice of solvent Preparation of solution Filtration of the solution Separating the crystals Drying of crystals Example Crystallisation of Phthalic acid 6. 2. SUBLIMATION Aim To separate volatile solids, which pass directly into vapour state on heating from a non-volatile solid. Principle A mixture of solid substances, such as camphor, benzoic acid, ammonium chloride, iodine etc., containing non-volatile substances, when heated, change directly into vapour without passing through the liquid state. Process 7. Fig :-Sublimation 8. 3. DISTILLATION Aim To separate a solution of a solid in a liquid and for separating a solution of two liquids whose boiling points are different. Principle Distillation involves the conversion of a liquid into its vapors upon heating and then cooling the vapors back into the liquid. Depending on the difference in boiling points of liquids. Types of distillation Simple Distillation Fractional Distillation Distillation Under Reduced...
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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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