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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
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∙ 13y ago
When selecting an HPLC column for method development, consider factors such as analyte characteristics (polarity, size, etc.), stationary phase chemistry (C18, phenyl, etc.), column dimensions (length, inner diameter), and particle size. Additionally, consider the mobile phase composition and operating conditions to optimize separation efficiency and resolution. Conduct preliminary scouting runs with different columns to determine the best fit for your analytes.
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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?
HPLC column volume refers to the total volume of the stationary phase in an HPLC column. It influences the separation efficiency and peak resolution of compounds in the chromatographic process. It is an important parameter to consider when optimizing HPLC methods for specific applications.
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 (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.
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 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'
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.
What is the HPLC method for related impurities in atorvastatin calcium tablets?
One common HPLC method for related impurities in atorvastatin calcium tablets involves using a C18 column with a mobile phase consisting of acetonitrile and buffer (e.g., ammonium acetate or phosphate buffer) at a specific pH. Detection can be done at a wavelength of around 240-245 nm. The method typically involves gradient elution to separate and quantify impurities present in the sample.
