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The resolution factor in HPLC is used to quantify the degree of separation between two adjacent peaks on a chromatogram. It is calculated by dividing the difference in retention times of the two peaks by the sum of their peak widths. A higher resolution factor indicates better separation between the peaks.
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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 (normal phase HPLC) separates compounds based on their polarity, where the stationary phase is polar and the mobile phase is nonpolar. RP-HPLC (reverse phase HPLC) separates compounds based on their hydrophobicity, where the stationary phase is nonpolar and the mobile phase is polar. RP-HPLC is more commonly used due to its versatility and ability to handle a wider range of compounds.
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 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.
What is dead volum in hplc?
Dead volume in HPLC refers to the volume in the system that is not actively involved in the separation process. It includes the volume of tubing, fittings, and detector cell that the mobile phase passes through without interacting with the stationary phase or analytes. Dead volume can lead to band broadening and decreased resolution in chromatographic separations.
How pH play an important role in hplc sepration?
depending on pH resolution is there
What is RRT and RRF in hplc?
In HPLC RRT means Relative Retention Time and RRF is Relative Response Factor
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.
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 (normal phase HPLC) separates compounds based on their polarity, where the stationary phase is polar and the mobile phase is nonpolar. RP-HPLC (reverse phase HPLC) separates compounds based on their hydrophobicity, where the stationary phase is nonpolar and the mobile phase is polar. RP-HPLC is more commonly used due to its versatility and ability to handle a wider range of compounds.
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.
Retention time calculation for hplc?
why RT was shifting & how to RT calculation in HPLC
