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What is an HPLC column?

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


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 you can select buffer in hplc according to pka value?

In HPLC, you can select a buffer based on its pKa value to achieve better separation of analytes by controlling pH of the mobile phase. Choose a buffer with a pKa value close to the desired pH for the separation, as this ensures the buffer will be most effective in maintaining stable pH. Selecting a buffer with a pKa within ± 1 unit of the desired pH is a commonly used guideline in HPLC method development.


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.


How buffer concentration effects on retention time in hplc method?

Buffer concentration can affect retention time in HPLC by influencing the pH of the mobile phase, which can in turn impact interactions between the analyte and stationary phase. Higher buffer concentrations can alter the ionization state of the analyte, leading to changes in its retention time. Additionally, buffer concentrations can also affect peak shape and resolution in the chromatogram.


How can analyze ibandronate sodium by hplc method?

column ----250mm 4.6mm L1 flow rate.--- 1ml/minute wavelength-- 195nmInjection Volume 10ul temperature ambent Buffer Preprations Take 600ml distilled water adjust pH 2.5 with phosphric acid take 500mg 1-pentane sulphonic sodium salt. add 100ml distilled watere after disolved of sodium salt Mobile Phase Ratio. Buffer 7: acetronitrile 3 (Buffer 700ml:Acetronitrile 300ml)


What types of buffer are used for hplc?

Common types of buffers used for HPLC include phosphate buffers, acetate buffers, citrate buffers, and ammonium acetate buffers. These buffers help to maintain the pH of the mobile phase, stabilize analytes, and provide consistent elution profiles. It's important to choose the right buffer based on the pH requirements of the analytes being analyzed.


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.


Why the pH of buffer is not changes in robustness of validation of hplc method?

it must change by (+- 0.3) to have control in pH meter error


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.