Electrophoresis Case Study

There are various techniques to separate a mixture of compounds from each other. One of the commonly used way to isolate compounds from a mixture of two compounds is called extraction. This method of extracting two compounds from each other relies on the different solubility of the compounds in two different solvents.

Me and my lab partner, obtained a mixture of a un known proportion from the instructor and then flow the guide line in our lab manual to separate the mixture by applying the separation method motioned in our lab manual pages 33-40 . In this experiment, the separation methods were decantation,

There are millions of different organic compounds. Most of them are found in mixtures and in order to achieve a pure form they need to be separated, isolated, and purified. However, there are endless numbers of possible mixtures, which make it impossible to have a pre-designed procedure for every mixture. So chemists often have to make their own procedures. The purpose of this experiment was to prepare the student to the real world by them designing their own procedure which will help them understand the techniques of separation and purification better. The goal was to extract two of the components of the

(a) (3 pts.) This video discusses 3 different types of chromatography. List each one mentioned, and describe their differences in as much detail as possible (your points earned will be proportional to the level of detail in your discussion). Which one was used in this lab demonstration?

Howlett, S. E. and Steiner, R. R. 2011. Validation of Thin Layer Chromatography with AccuTOF-DART™ Detection for Forensic Drug Analysis*. Forensic Sciences [e-journal] 56 (5), pp. 1261--1267. Available through: Anglia Ruskin University Library website http://libweb.anglia.ac.uk [Accessed on 11 March 2014].

The purpose of this experiment is to observe how chromatography can be used to separate mixtures of chemical substances. Chromatography serves mainly as a tool for

     Conventional Liquid Chromatography is most commonly used in preparative scale work to purify and isolate some components of a mixture. It’s also used in ultra trace separations where small disposable columns are used once and then discarded. Analytical separations of solutions for detection or quantification typically use more sophisticated high-pressure liquid chromatography instruments.

Thin layer chromatography (TLC) separates compounds, through migration, from a mixed solution with the assistance of a solvent and an absorbent strip of cellulose. The purpose of this lab is to allow students to determine an unknown amino acid by comparing results to six known amino acids (slowest to fastest: lysine, glycine, alanine, tyrosine, phenylalanine, and leucine) and properly separate a combined solution of amino acids using the TLC method. But to fully understand the lab, two terms must be understood: amino acid and protein. Amino acids are building blocks of proteins linked by peptide bonds that contain both carboxyl and amino functional groups.

Gel Electrophoresis is used to separate macromolecules like DNA and RNA by their size, or proteins by their charge and size. In this experiment, I used the Gel electrophoresis to determine the presence of all dyes in a specific dyes mixture.

Further investigate and determine its identity. By utilizing chromatography, a forensic analyst can identify each material and/or chemical that were meshed together to form the sample under investigation. The identification can be determined once the sample is placed in a liquid or when absorbed on a solid surface, and the separation of the molecules becomes either mobile or stationary. This basic analytical technique is beneficial with classifying drugs that has been collected from a crime scene and transported to a crime laboratory for analyses. There are four main variations of chromatography, however, Gas Chromatography is the most widely used technique (Woodford, 2017). Gas chromatography is a valuable method in forensic science because

The purpose of the lab was to separate a mixture using gas chromatography and analyze the produced chromatogram. Also, the effect of temperature variance of the Mini GC on the chromatogram was analyzed and the best temperature-pressure profile was determined based on the results.

When raising the pH of mobile phase buffer will cause the molecule become less protonated and hence less positively charged. The result is that the protein no longer can form an ionic interaction with the negatively charged solid support, which ultimately results in the molecule to elute from the column. Phenylalanine eluted fastest (pI=5.48) because it is lower than pH6. Leucine and Alanine was eluted next in pH6 because of their pI number 5.98 and 6.00, respectively. The next to elute out in pH11 buffer was Lysine. It has highest pI number (pI=9.74) and it will lose its charge at pH 11. Refer to figure 3, you can see phenylalanine and alanine showed up at pH 6 and pH 3, there was an error at pH 3 because the amino acid should not show up here based on their isoelectric point close to 6. The pH must be between the pI or higher in order for it to elute. The error maybe due the contamination while performing the experiment or can be not properly performed. Also, we can see that the lysine remained positively charge until it gets to pH 11 because the pI number is 9.74. On figure 3, it also showed phenylalanine at pH 11. This could be the amino acids were still in contact with the Dowex resin and Dowex was not

Nevertheless, GC still plays an important role in certain types of quantitative analysis and has broad application in qualitative analysis. The advantages of GC may be overlooked since HPLC is dominating the quantitative analysis in pharmaceutical industry nowadays. Capillary GC is capable of performing much more efficient separations than HPLC with the limitation that the compounds being analyzed must be volatile or must be rendered volatile by formation of suitable derivative and must also be thermally stable. GC is widely used in environmental science, brewing, the food industry, perfumery and flavourings analysis, the petrochemical industry, microbiological analyses and clinical biochemistry. (R. Kellner,

1. Gas chromatography is a type of chromatography that allows for separation and analysis of a compound. It does this by vaporizing the sample then using a carrier gas to push it through a column which then separates it based on an equilibrium formed between solution and dissolution.

AIM : Thin-Layer Chromatography can show many different characteristics of a mixture. It is recognized for isolation , separation ,identification, and anaylsis of the mixture’s components. The purpose of this experiment is to separate carbohydrates into its pure components such as mixtures of monosacrides by TLC. TLC is used to identify sugars in normal and pancreatic disease urine, the procedure is easy and reproducible .