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What are the advantages of UPLC over HPLC?
UPLC (Ultra Performance Liquid Chromatography) typically provides faster analysis times, higher resolution, and improved sensitivity compared to traditional HPLC (High Performance Liquid Chromatography). UPLC systems use smaller particle sizes in stationary phases and higher pressures, leading to better separation efficiency and lower solvent consumption.
How do you distinguised np-hplc and rp-hplc?
NP-HPLC is "Normal Phase" HPLC, wherein the solvents used are less polar than the substrate in the HPLC column (e.g. using hexane or dichloromethane with a silica HPLC column). RP-HPLC is "Reverse-Phase" HPLC, wherein the solvents used are more polar than the substrate in the HPLC column (e.g. using Water and Methanol with a octadecylsilane (ODS or C18) column).
How do you calculate concentration from peak area in HPLC analysis?
To calculate concentration from peak area in HPLC analysis, you can use the formula: Concentration Peak Area / (Slope x Injection Volume). The peak area is obtained from the chromatogram, the slope is the calibration curve slope, and the injection volume is the volume of the sample injected into the HPLC system.
What is mean by delay volume in HPLC analysis?
Delay volume in HPLC analysis refers to the volume of liquid in the system that is not actively participating in the separation process. It includes the volume of tubing, fittings, and the void volume of the column. Minimizing the delay volume is important for maintaining good chromatographic resolution and reducing analysis time.
Why caffeine is used in calibration of hplc?
Caffeine is used as a calibration standard in HPLC because it is a readily available, stable compound with known retention times and peak shapes. Its use allows for the accurate determination of column efficiency, resolution, and peak symmetry, making it a valuable compound for calibration purposes in HPLC.
What is RRT and RRF in hplc?
In HPLC RRT means Relative Retention Time and RRF is Relative Response Factor
How pH play an important role in hplc sepration?
depending on pH resolution is there
What is the use of calculating asymmetry factor in hplc?
The asymmetry factor in HPLC is used to assess the peak shape of a chromatographic peak. It is calculated by dividing the front part of the peak by the back part, providing information on the peak tailing or fronting. A symmetrical peak typically has an asymmetry factor close to 1, indicating good peak shape.
Why chiral hplc methods are non aqueous?
Chiral HPLC methods are often non-aqueous because many chiral stationary phases are not compatible with high levels of water due to stability and performance issues. Using non-aqueous solvents can also improve the resolution and selectivity of chiral separations in HPLC.
What are the HPLC Calibration parameters and elaborate it?
1. Flow rate 2. Temp. of column 3. Detector function 4. Resolution
Is resolution the most important factor when choosing an LCD monitor?
resolution
Peak-to-valley ratio in HPLC?
The peak-to-valley ratio in high-performance liquid chromatography (HPLC) is a measure of the separation between the highest peak and the adjacent valleys in a chromatogram. It is calculated by dividing the peak height by the lowest valley height around the peak. A higher peak-to-valley ratio indicates better resolution and a more efficient separation of analytes.
Why is column revers phase in hplc?
Column reverse phase in High-Performance Liquid Chromatography (HPLC) refers to the use of a nonpolar stationary phase and a polar mobile phase. This configuration allows for the separation of compounds based on their hydrophobicity, where more hydrophobic compounds interact more strongly with the stationary phase and elute later. Reverse phase HPLC is widely used because it provides high resolution and sensitivity for a variety of analytes, making it suitable for pharmaceuticals, environmental samples, and biological fluids.
What are the advantages of UPLC over HPLC?
UPLC (Ultra Performance Liquid Chromatography) typically provides faster analysis times, higher resolution, and improved sensitivity compared to traditional HPLC (High Performance Liquid Chromatography). UPLC systems use smaller particle sizes in stationary phases and higher pressures, leading to better separation efficiency and lower solvent consumption.
What is difference between high pressure liquid chromatography and high performance lequid chromatography?
High pressure liquid chromatography (HPLC) and high performance liquid chromatography (HPLC) are often used interchangeably. HPLC refers to modern liquid chromatography systems with high resolution and efficiency, while high pressure liquid chromatography specifically highlights the use of higher pressures in the system to improve separation and speed. Both terms generally refer to the same chromatographic technique.
How do you distinguised np-hplc and rp-hplc?
NP-HPLC is "Normal Phase" HPLC, wherein the solvents used are less polar than the substrate in the HPLC column (e.g. using hexane or dichloromethane with a silica HPLC column). RP-HPLC is "Reverse-Phase" HPLC, wherein the solvents used are more polar than the substrate in the HPLC column (e.g. using Water and Methanol with a octadecylsilane (ODS or C18) column).
Retention time calculation for hplc?
why RT was shifting & how to RT calculation in HPLC
