HILIC uses a polar stationary phase and is ideal for separating polar compounds (e.g., sugars, amino acids). RPC uses a non-polar stationary phase and is best for non-polar compounds (e.g., lipids, hydrophobic drugs). HILIC relies on hydrophilic interactions, while RPC relies on hydrophobic interactions. Both differ in their mobile phases: HILIC uses polar solvents, while RPC uses less polar or organic solvents.
BT21CME111 Shreya Dhote Could you go into more detail about how the mobile phase composition influences separation in HILIC, particularly with polar analytes?
The mobile phase composition in HILIC significantly influences separation, especially for polar analytes. Higher water content enhances polar interactions, increasing retention, while higher organic solvent content (like acetonitrile) decreases retention by weakening these interactions. Buffers and pH also play a role in ionizing analytes and ensuring optimal interaction with the stationary phase. Balancing these factors is key to achieving good separation and resolution in HILIC.
What is the advantage of HILIC over reverse-phase chromatography?
HILIC is better suited for polar and hydrophilic compounds, which often have poor retention in reverse-phase systems.
BT21CME047 Harshita
Difference between HILIC and Reverse phase Chromatography
HILIC uses a polar stationary phase and is ideal for separating polar compounds (e.g., sugars, amino acids). RPC uses a non-polar stationary phase and is best for non-polar compounds (e.g., lipids, hydrophobic drugs). HILIC relies on hydrophilic interactions, while RPC relies on hydrophobic interactions. Both differ in their mobile phases: HILIC uses polar solvents, while RPC uses less polar or organic solvents.
BT21CME026
What types of compounds are analyzed using HILIC?
Polar compounds, including sugars, amino acids, nucleotides, and small polar metabolites are analyzed using HILIC
BT21CME043 How is retention time affected in HILIC?
Thankyou Devendra for asking this question, in HILIC retention increases with higher water content in the mobile phase.
BT21CME111 Shreya Dhote
Could you go into more detail about how the mobile phase composition influences separation in HILIC, particularly with polar analytes?
The mobile phase composition in HILIC significantly influences separation, especially for polar analytes. Higher water content enhances polar interactions, increasing retention, while higher organic solvent content (like acetonitrile) decreases retention by weakening these interactions. Buffers and pH also play a role in ionizing analytes and ensuring optimal interaction with the stationary phase. Balancing these factors is key to achieving good separation and resolution in HILIC.