Chromatography lecture and my learning diaries
The chromatography is a collective terms for separation of mixtures
techniques. The sample (Mixtures) is dissolved in the mobile phase,
which carried it through the stationary phase. The separation was
achieved by the difference of travel time/speeds. Several chromatography
methods have been developed during the past time including paper
chromatography, gas chromatography, and high performance liquid
chromatography (HPLC). In this lecture, the teacher mainly introduced
the principle of HPLC.
HPLC is a technique to separate a mixture of compounds in analytical
chemistry and biochemistry with the purpose of identifying, quantifying
and purifying the individual component. The instruments consists of PUMP
(moves the mobile phase and sample through the column), injector (add
samples), column and detector.
Different liquid chromatography including gel filtration, ion exchange,
affinity and reversed phase chromatography are utilized according to the
protein/peptide properties including size, net surface charge,
hydrophobicity, respectively. Different methods can be combined
according to the purpose, but the most important is keep it simple
because quite a lot of samples are lost during the process.
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