You are the manager in a laboratory where you develope methods that are useful i

Question

You are the manager in a laboratory where you develope methods that are useful in metabolomics (study of metabolites) and proteomics (study of proteins), meaning that you need to study both relatively small (tens to hundreds of Da) to large (thousands of Da) molecules, and you are tasked with buying analytical instumentation to do these analyses. In the method that you are developing, you would like to qualitatively and quantitatively study metabolites and proteins from human body fluids (mixture of many compounds in a complex matrix). Although money is not a big concern, you would like to develope a method that would keep money in mind as well as do the analysis as quickly as possible. Only a small subset of analytes is electroactive or have chromophores. Analyte amounts can vary from pmol to mmol, and you would like to minimize the amount of body fluids you must obtain. Most of the compounds have limited or no volatility up to 300 degrees C.

a. What technique(s) would you buy and why? "Why" should answer how the technique(s) meet or take care of analytical constraints.

b. In as much detail as you can, outline the instrumentation that you would use with your techniques. If more than one is used, specify the order.

c. From these techniques how do you identify your components (obtain qualitative data)?

Explanation / Answer

Ans a) NMR (Nuclear magnetic Resonance) is one of the techniques or instrument that can be purchased for studying the process of metabolomics and proteomics. It helps in analysing multi-component mixtures as sample preparation required for analysis is important. It is a rapid as well as non-destructive process and is integrated with advanced technology for studying the genomics and proteins. It is highly sensitive when it comes to studying the complex metabolites. It is used for studying and analysing as it helps in observation of the local magnetic fields that is present around the nuclei to understand the structure. However there are some constraints that are observed while using NMR. The main constraint is incorporating or inputting the structure general programs. Each of the structure needs to put into separate set.

Ans b) It is an analytical method which helps in analysing the content and the purity of the material used. It helps in quantitative analyse of mixture. The principle of the NMR is that it is identified based on the nuclei around the spin and the electrical charged. In this the energy is transferred from base to higher level helping in identifying frequency and the energy emitted. The principle that all the nuclei are electrically charged and on application of magnetic field, there is transfer of energy which helps in identification of the protein.

Ans c) The quantification of the protein or the component from the mixture is done by dispersion. The separation can be done on the base of the frequency of the energy. Every component has its own frequency and wavelength which helps in analysing and quantifying the compound.

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