HPLC Chromatography Basics Explained

(You can also watch new HPLC video with demonstration here - 👉 • HPLC Chromatography De... )
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All you need to know about the HPLC Chromatography!
HPLC(High-performance liquid chromatography) also referred as high-pressure liquid chromatography, is a technique used in analytical labs to separate, identify, and quantify each component in a mixture.HPLC separates and purifies compounds according to their polarity.
I have tried to simplify the whole process of HPLC Chromatography and explain in simple terms. Let’s look at the main components involved in HPLC in this video.
Later in this video, I discussed about Normal Phase Chromatography, Reverse Phase Chromatography and their comparison and differences.
Watch this video for more details.
Other Video Links:
ER TO GOLGI Transport: • Endoplasmic Reticulum ...
KDEL and KKXX Sequence: • KDEL and KKXX Signal, ...
If you have any doubts: Please leave a comment in the comments section below.
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Thank you very much for your presentation. Would like to know about the buffer you mentioned to equilibrate before the sample run (getting the base line). can you explain more about the equilibration and the buffers to use thank you!

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Hi mam, I want to quantify triphenylantimony from my sample, for that which mobile phase n which column is needed?

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[@namitadhiman6756]

Mam so the main principle of HPLC technique will be this or something else. The separation principle of HPLC is based on the distribution of the analyte (sample) between a mobile phase (eluent) and a stationary phase (packing material of the column). Depending on the chemical structure of the analyte, the molecules are retarded while passing the stationary phase. The specific intermolecular interactions between the molecules of a sample and the packing material define their time “on-column”. Hence, different constituents of a sample are eluted at different times. Thereby, the separation of the sample ingredients is achieved. A detection unit (e.g. UV detector) recognizes the analytes after leaving the column. The signals are converted and recorded by a data management system (computer software) and then shown in a chromatogram. After passing the detector unit, the mobile phase can be subjected to additional detector units, a fraction collection unit or to the waste. In general, a HPLC system contains the following modules: a solvent reservoir, a pump, an injection valve, a column, a detector unit and a data processing unit (Fig. 1). The solvent (eluent) is delivered by the pump at high pressure and constant speed through the system. To keep the drift and noise of the detector signal as low as possible, a constant and pulseless flow from the pump is crucial. The analyte (sample) is provided to the eluent by the injection valve.

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That was simple way of explaining...Pls explain us about Method development by HPLC,it's process etc..

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So finally how can we know that the different component is separate ? and how we identify the component ?

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Very nice mam almost done hplc

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Can you tell me please ! Question What is advantage of reverse phase HPLC over normal phase HPLC ?

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Reply me

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