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firs you mist know the polarity for sample, wen the sample polar you can use "RP" column like C18 or C8 ( C18 first in pharmaceutical) . wen sample non polar use "NP" column like silica or CN Column.
after that you can change the column in same packing to solve tailing, retention time, Resolution..... or any problem by change column length, particle size or carbon loud
What else can I help you with?
Why is anthracene is used for calibration of hplc?
Anthracene is used as a calibration standard in High Performance Liquid Chromatography (HPLC) because it has a well-defined retention time and peaks in the UV-visible spectrum, making it easy to detect and quantify. Its consistent behavior helps in determining retention times, resolving power, and column efficiency during method development and troubleshooting in HPLC.
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.
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.
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 the definition of method development by hplc?
Method development is a process amenable to continuous improvement
What are the guidelines for method development of hplc according to ich?
the same guidelines for method validation
Why is anthracene is used for calibration of hplc?
Anthracene is used as a calibration standard in High Performance Liquid Chromatography (HPLC) because it has a well-defined retention time and peaks in the UV-visible spectrum, making it easy to detect and quantify. Its consistent behavior helps in determining retention times, resolving power, and column efficiency during method development and troubleshooting in HPLC.
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 is rs-hplc?
"RS-HPLC method" means "Related Substance HPLC Method".
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
What has the author Sukwinder Kaur Gill written?
Sukwinder Kaur Gill has written: 'Development of a method for the determination of chlortetracycline (CTC) in a three component mixture (Microfac) by HPLC'
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.
