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HPLC Column is one type of tube containing a stationary phase react with mobile phase to detect peak

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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).


What can one read in the HPLC Column?

In an HPLC column one can see very small molecules such as ATP, histidine, glucose, uracil, and pyridine. It is a form high quality of liquid Chromatography.


What happen after used dipotassium buffer ph 9 on hplc column?

Nothing will happen


Retention time in Hplc?

Retention time in High Performance Liquid Chromatography (HPLC) refers to the time it takes for a compound to travel through the chromatography column and elute from the detector. It is a key parameter for identifying and characterizing compounds in a sample. Retention time is influenced by factors such as the column type, mobile phase composition, and compound properties.


Does HPLC can read histamine and TVB-N?

Yes, HPLC can be used to analyze histamine and TVB-N (Total Volatile Basic Nitrogen) in food samples. HPLC is a sensitive and selective technique that can separate and quantify various compounds, including histamine and TVB-N, based on their chemical properties. Pre-column derivatization may be required for some compounds to enhance their detection sensitivity in HPLC analysis.


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.


Can you use a C18 column for HPLC with fluorescence detector and methanol as the mobile phase?

Yes, you can use a C18 column and methanol as a mobile phase with fluorescence detector. Fluorescence detector is generally used as it can detect the presence of compounds at a very low concentration.


What is the function of pump in HPLC?

A high-performance liquid chromatography, or HPLC, refers to a technique in analytic chemistry that is used to separate the components in a mixture. The pump in HPLC passes a pressurized liquid solvent that contains the sample mixture through a column filled with a solid adsorbent material.


Definition of standard in HPLC?

In HPLC, a standard is a known compound with a defined chemical structure and purity used for comparison and identification purposes. Standards are essential for calibrating instruments, determining retention times, and quantifying unknown compounds in samples during analysis.


What is negative peak in hplc?

In HPLC, the negative peak refers to a trough or valley observed in the chromatogram where the signal intensity drops below the baseline. This can occur due to factors such as noise, interference, or improper column packing. Negative peaks can sometimes affect the accuracy and precision of peak integration and quantification in HPLC analysis.


Can C18 column for HPLC be used with UV and visible spectra detector and Acetonitril as mobile phase?

Of course.... there isn´t problem....


What are the HPLC Calibration parameters and elaborate it?

1. Flow rate 2. Temp. of column 3. Detector function 4. Resolution