using cation exchange solid phase extraction‚ repeated extraction with 10 mM formic acid solution and precipitation at low temperature by centrifugation. The utilise of C-18 reversed-phase columns were slightly more polar performed to achieve a better separation of acrylamide. Eluent system of liquid chromatography used a time programme gradient in order to obtain a better selectivity. The mobile phase was a mixture of 10 mM formic acid and methanol. The performance parameters of precision‚ accuracy
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SAMPLE PREPARATION METHODS FOR LIQUID SAMPLES Table of Contents SAMPLE PREPARATON Analytical procedures consist of numerous stages‚ the most important of which is the collection of a sample and its preparation for analysis as samples are usually not in a suitable form for direct introduction into analytical instruments (Tankiewicz et al.‚ 2011). Sample preparation can be thought of as any treatment that the sample is subjected to following its collection‚ prior to its analysis
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Connor Lauffenburger 3/17/13 pGlo Transformation Lab Report I Introduction The purpose of this experiment was to show the genetic transformation of E. coli bacteria with a plasmid that codes for Green Fluorescent Protein (GFP) and contains a gene regulatory system that confers ampicillin resistance. A plasmid is a genetic structure in a cell that can replicate independently of chromosomes. In this lab‚ the Green Fluorescent Protein‚ which is typically found in the bioluminescent jellyfish Aequorea
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fermentation broth by various chromatography and analysis using RP HPLC techniques- specializations on lipstatin drug. Abstract: The main focus area of this project is to purify the microbe derived drug lipstatin by using various trials of chromatographic columns with different packing materials or varying mobile phase composition or altering the concentration or volume of the load of the crude sample‚ which has to be purified. Till date I have done trials with silica gel columns. I propose in future to
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Prior to use‚ the column was equilibrated using 20mM sodium phosphate buffer (pH6.5). The column was washed for 10 CV with the same buffer after the protein loading and the bound proteins were eluted with a linear gradient of NaCl (0-0.5M) in the same buffer over 10 CV at a flow rate of 1mL/min. The negative
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different chemical properties of each compound. We will accomplish this by a separation procedure known as distillation‚ which relies on each compound having a distinct and separate boiling point. Our pure products will be analyzed with gas chromatography to determine the success of the distillation. Procedures The experiment was performed as stated in the course textbook: Pavia‚ D. L.‚ Lampman‚ G. M.‚ Kriz‚ G. S.‚ Engel‚ R. G. Introduction to Organic Laboratory Techniques: A Microscale Approach
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degradation products. The chromatographic conditions were optimized to achieve the highest performance parameters using Zorbax Eclipse Plus C18 rapid resolution column (4.6 X 100 mm‚ 3.5 µm)‚ with a mobile phase composed of 72.5 % acetate buffer pH 5.5 and 27.5 % ethanol‚ flowing at 1 ml min-1. The Diode Array Detector (DAD) was set at 315 nm and the column oven was adjusted at 45 °C. The method was validated according to the ICH guidelines with respect to system suitability‚ specificity‚
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300 microliter predictor columns often give higher recoveries than the Tricorn columns‚ with the minimum value of 66% recovery‚ even giving good recoveries using smaller salt concentrations and buffer ionic strength‚ which Tricorn cannot‚ particularly struggling during pH 5.75‚ when almost a third of the contour plots gave recoveries 40% or below. In addition‚ there were more samples taken when using the Predictor columns (9 samples per contour plot)‚ with Tricorn columns only using 5 samples per
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supernatant is to be removed and labeled accordingly in a separate vessel. Supernatant is the liquid above the sediment after a solution has been run through a centrifuge. The now centrifuged solution should be ran through a device that achieves column chromatography‚ such as an automated fraction collector. These devices sort a solution based on particle size. Other methods to extract proteins (which can be many different
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procedures of this laboratory will be used to gain a conceptual and practical understanding of separation techniques used to isolate monomers from their respective polymers. Namely‚ the techniques that will be explored are dialysis and gel filtration chromatography. In the remaining two experimental procedures‚ colorimetric assays will be used to detect the presence of certain carbohydrates. Glucose oxidase and Iodine Reactions will be performed in conjunction to demonstrate the procedure of such assays
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