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Why water is not used in normal phase hplc analysis?
In normal-phase chromatography, the stationary phase is polar and the mobile phase is a mixture of non-polar solvents such as hexane and slightly more polar solvents such as isopropanol. water is the most polar solvent of all solvents. If you use water as a mobile phase, the polar analytes will remain dissolved in water and there will be no retention of analytes on the stationary phase. If there is no retention on stationary phase, there is no separation
Why the retention time will decrease if the polarity of mobile phase increase in normal phase HPLC?
because in normal phase HPLC mobile phase is non polar and stationary phase is polar. Most of the compound of interest are polar, if you increase the polarity of mobile phase compound of analyte will stay in mobile phase and will elute faster and retention time will be shorter.
How do you estimate Serratiopeptidase in Pharmaceutical products Is there a HPLC method available?
To estimate Serratiopeptidase in pharmaceutical products, HPLC (High-Performance Liquid Chromatography) is a commonly used method. A validated HPLC method can separate and quantify Serratiopeptidase in a sample, providing accurate results for quality control purposes in pharmaceutical analysis. It is essential to use appropriate standards and optimize chromatographic conditions for this analysis.
What is difference between isocratic and gradient hplc?
In isocratic HPLC, the mobile phase composition remains constant throughout the entire run, leading to constant elution times for all analytes. In gradient HPLC, the mobile phase composition is changed during the run, allowing for better separation of complex mixtures by adjusting the solvent strength over time.
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 key differences between reverse phase and normal phase HPLC techniques?
Reverse phase and normal phase HPLC techniques differ primarily in the polarity of the stationary phase and mobile phase. In reverse phase HPLC, the stationary phase is nonpolar and the mobile phase is polar, while in normal phase HPLC, the stationary phase is polar and the mobile phase is nonpolar. This polarity difference affects the retention and separation of compounds in the sample.
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.
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 an HPLC column?
HPLC Column is one type of tube containing a stationary phase react with mobile phase to detect peak
What are the key differences between HPLC reverse phase and normal phase chromatography techniques?
In reverse phase HPLC, the stationary phase is nonpolar and the mobile phase is polar, while in normal phase HPLC, the stationary phase is polar and the mobile phase is nonpolar. This difference in polarity affects how compounds interact with the stationary phase, leading to variations in separation and elution times.
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 are the key differences between HPLC normal phase and reverse phase chromatography techniques?
In normal phase HPLC, the stationary phase is polar and the mobile phase is nonpolar, while in reverse phase HPLC, the stationary phase is nonpolar and the mobile phase is polar. This difference in polarity affects how compounds interact with the stationary phase, leading to different separation mechanisms and selectivity in each technique.
Why water is not used in normal phase hplc analysis?
In normal-phase chromatography, the stationary phase is polar and the mobile phase is a mixture of non-polar solvents such as hexane and slightly more polar solvents such as isopropanol. water is the most polar solvent of all solvents. If you use water as a mobile phase, the polar analytes will remain dissolved in water and there will be no retention of analytes on the stationary phase. If there is no retention on stationary phase, there is no separation
What are the differences between reverse phase HPLC and normal phase chromatography techniques?
Reverse phase HPLC and normal phase chromatography are two types of chromatography techniques that differ in the polarity of the stationary phase and mobile phase. In reverse phase HPLC, the stationary phase is non-polar and the mobile phase is polar, while in normal phase chromatography, the stationary phase is polar and the mobile phase is non-polar. This difference in polarity affects the separation of compounds based on their interactions with the stationary phase, leading to different retention times and selectivity in each technique.
Why the retention time will decrease if the polarity of mobile phase increase in normal phase HPLC?
because in normal phase HPLC mobile phase is non polar and stationary phase is polar. Most of the compound of interest are polar, if you increase the polarity of mobile phase compound of analyte will stay in mobile phase and will elute faster and retention time will be shorter.
How you can analyze pregabalin on HPLC with UV detector?
We can quantitatively analyse pregabalin on hplc with uv detector, wavelength will be 210 n.m. and mobile phase will be 5 % acetonitrile. standard & sample solution preparation should be in mobile phase.
What is difference bet GLC and HPLC?
GLC has a stationary liquid phase and gas moving phase HPLC had a stationary solid phase and liquid moving phase HPLC is done under high pressure. HPLC can be used for thermally unstable compounds as opposed to GLC HPLC can be used for polar or low volatile compounds as opposed to GLC
