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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 volume HPLC?
The dead volume in HPLC is 137.45. The dead volume in science is used in retention measurements and also in thermodynamic studies and the abbreviation HPLC stands for High Pressure Liquid Chromatography.
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 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 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
What type of species can be separated by HPLC but not by gas liquid chromatography?
mixture of enantiomers can be separated by HPLC
What is hplc column volume?
Since an HPLC column is a cylinder, the simplest estimate for the column volume is the equation V=L*pi*r2, where L = length of column (typically 50-250 mm, or 5-25 cm), and r=radius of the column, where typical internal diameters of HPLC columns are 2.1 mm, 3 mm, 4 mm, and 4.6 mm. For example, suppose you have a column that is 25 cm long by 4.6 mm internal diameter (ID). Since the ID is in mm, you first convert to cm, then divide by 2 to get 0.23 cm radius. The column volume equation then is: V = 25 * pi * (0.23)2 = 25 * pi * 0.0529 = 1.3225 * pi = 4.15 cm3 From there, you can convert cm3 to mL directly, so your column has a volume of 4.15 mL. However, you must also allow for the relative porosity of the packing material in your column, which is harder to measure. Typically, an unretained analyte will be injected through the column at a known flow rate, and the time it takes for the analyte to exit the column is used to determine a better approximation of column volume. In the case of using an unretained analyte (which in reversed-phase HPLC, the analyte might be Uracil), using the same 25 cm by 4.6 mm column above and a 1 mL/min flow rate, suppose the analyte elutes from the column at 3.2 minutes. The column volume would then be 3.2 minutes * 1.0 mL/min = 3.2 mL, which does not agree with the calculated column volume. This is due to the fact that the particles in the column take up some of the volume of the column, so the total column volume is reduced by the amount of space they take up.
What is rs-hplc?
"RS-HPLC method" means "Related Substance HPLC Method".
Where can one purchase HPLC detectors?
You can purchase used HPLC detectors and other equipment from the usedhplc website or from the ebay bidding website. Alternatively you can buy HPLC detectors from the equipnet website.
What is RRT and RRF in hplc?
In HPLC RRT means Relative Retention Time and RRF is Relative Response Factor
What are the key differences between normal phase HPLC and reverse phase HPLC in terms of their separation mechanisms and applications?
Normal phase HPLC separates compounds based on their polarity, with the stationary phase being polar and the mobile phase being nonpolar. Reverse phase HPLC separates compounds based on their hydrophobicity, with the stationary phase being nonpolar and the mobile phase being polar. Normal phase HPLC is typically used for separating polar compounds, while reverse phase HPLC is used for separating nonpolar compounds.
