Pretend you need to analyze solutions that contain variable kinds and amounts of amino acids|My homework helper

Posted: March 5th, 2023

Pretend you need to analyze solutions that contain variable kinds and amounts of amino acids ( not whole proteins or peptides ). To do this analysis , you need a practical method that can separate and quantitatively detect all twenty fundamental – amino acids . Briefly discuss one procedure , other than paper chromatography / ninhydrin , that could be used to solve this analytical problem . It’s okay if your procedure requires more than one step ( ie a separation technique followed by an analytical step ).

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a ) a description of the molecular basis for the separation chemistry

b ) a description of the technique that will be used for detection of the analytes and how /why detection would be quantitative

c ) the strategy that will be used to characterize which of the twenty amino acids were in a given mixture

d ) the limitations of the procedure especially regarding whether it would really work for all 20 amino acids and whether it would truly be quantitative . ( for example , if you think it might not separate every amino acid, explain specifically which ones might be left unseparated and why).

e ) the peer- reviewed reference for your chosen technique (bear in mind that an “open source” reference such as Wikipedia does not count as a peer-reviewed reference)

 

SOLUTION

One procedure that could be used to separate and quantitatively detect all twenty amino acids is high-performance liquid chromatography (HPLC) coupled with fluorescence detection. ) HPLC separates molecules based on their differential affinities for a stationary phase and a mobile phase. In this case, a reversed-phase HPLC column would be used, where the stationary phase is nonpolar (e.g., C18) and the mobile phase is polar (e.g., an aqueous buffer containing organic modifiers such as acetonitrile or methanol). Amino acids differ in their hydrophobicity and polarity, which affects their interactions with the stationary and mobile phases, leading to differential retention times and separation.

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