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How do you choose a stationary phase?

How do you choose a stationary phase?

In other words, you should choose a stationary phase with similar polarity characteristics to those of the sample components. For example, higher polarity of the stationary phase will increase the selectivity of the cartridge for the polar components in the sample.

What stationary phase is used in HPLC?

The typical stationary phase for HPLC can be silica-based or polymer-based (native or synthetic). Once filled into the column it does not shift. Due to the polarity they are divided into reversed stationary phases and normal stationary phases.

How are mobile phase chosen in HPLC?

The nature of the carrier (solvent) for the analytes can be chosen during the sample preparation to satisfy specific requirements such as volatility and miscibility with the mobile phase in HPLC. In reverse phase columns, the mobile phase is more polar than the stationary phase.

What is C18 stationary phase?

C18 stationary phases are not identical from one C18 HPLC column to another. C18 simply means that the molecules contain 18 carbon atoms, so the other atoms in the molecule can vary, leading to significantly different substances.

What is selectivity in HPLC?

Selectivity is the ability of an HPLC method to separate two analytes from each other. Selectivity usually is abbreviated with the Greek letter α, and is calculated as: α = k2 / k1 where k1 and k2 are the retention factors, k, of the first and second peaks of a peak pair.

How can we increase selectivity in HPLC?

Often the simplest and most effective way to improve your chromatographic results is to change your column’s phase or solid support.

What is a stationary phase?

Stationary phase is the stage when growth ceases but cells remain metabolically active. Several physical and molecular changes take place during this stage that makes them interesting to explore. The characteristic proteins synthesized in the stationary phase are indispensable as they confer viability to the bacteria.

Does HPLC use polar stationary phase?

The combination of a polar stationary phase and a nonpolar mobile phase is called normal-phase chromatography. In reversed-phase chromatography, which is the more common form of HPLC, the stationary phase is nonpolar and the mobile phase is polar.

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