Which solvent is used in reverse phase HPLC?

The solvents must be miscible with water, and the most common organic solvents used are acetonitrile, methanol, and tetrahydrofuran (THF). Other solvents can be used such as ethanol or 2-propanol (isopropyl alcohol).

What are the common solvents used in TLC?

Solvent Systems for Thin Layer Chromatography

• Polar compounds: 100% EtOAc or 5% MeOH/dichloromethane.
• Normal compounds: 10-50% EtOAc/Hexane.
• Nonpolar compounds: 5% EtOAc/hexane, 5% ether/hexane, 100% hexane.

How do you reverse phase chromatography?

Reverse-phase HPLC involves binding an organic molecule to a stationary phase, often silica derivatized with alkyl chains, in a relatively polar environment (the mobile phase), which could contain water, and then eluting the organic molecule using a gradient of a less polar organic solvent.

Why acetonitrile is used as solvent in HPLC?

Acetonitrile is often used because of its low UV cutoff, lower viscosity (methanol forms highly viscous mixtures with water at certain concentrations), and higher boiling point.

Which are polar solvents?

Polar solvents have a “positive” and a “negative” charge at different places in their structures and will dissolve other polar substances. Water is a polar solvent; other polar solvents include acetone, acetonitrile, dimethylformamide (DMF), dimelthylsulfoxide (DMSO), isopropanol, and methanol.

Why is ethyl acetate and hexane used in TLC?

In TLC, the separation of mixture is based on polarity between stationary phase and mobile phase. Ethyl acetate has the polar property whilst hexane has the non polar property. Both can be used as mobile phase in TLC where the solubility may also affect the migration of the seperated substances.

What is reversed phase liquid chromatography?

Reversed phase liquid chromatography (RPLC) is a mode of high-performance liquid chromatography (HPLC) that employs a nonpolar stationary phase (most frequently a hydrocarbon chain chemically bonded to porous silica particles) and a polar mobile phase constituted by water and at least a water-miscible organic solvent.

How do you convert reverse phase to normal phase?

To switch from reversed- to normal-phase – prime with methanol (or acetonitrile) followed by ethyl acetate and then hexane, all between 50 to 100 mL. For most flash systems, the above solvent switching is a manual operation.

Why is methanol used in HPLC?

Methanol is a polar-protic solvent, whereas acetonitrile is a polar-aprotic solvent and possesses a stronger dipole moment. This means that the organic modifier used in the mobile phase can have a powerful effect on chromatographic selectivity.

What are the solvents used in HPLC?

Normal Phase HPLC separates the analytes using their varying degree of affinity to the phase which is polar by nature. Non-polar solvents like chloroform, cyclohexane, benzene, carbon tetrachloride etc. are used.