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HPLC - High Performance Liquid Chromatography
Principle of hplc -
HPLC seperates mixture of compounds on the basis of polarity.
{Polarity refers to : the greater the difference in electron affinity i.e. electronegativity between atoms in a covalent bond, the more polar the bond.partial negative charges are found on the most electronegative atoms, the others are partially positive.The moleacular electrostatic potential energy of a hydrogen ion at a particular location near a molecule. Negative electrostatic potential corresponds to : partial negative charges , Positive electrostatic potential corresponds to : partial positive charges.}
It is used to analyze,identify,purify and quantify componds.
It has a mobile phase,a stationary phase,detector.The mobile phase is continuously pumped at a fixed flowrate through the system and mixed by the pump.The injector is used to introduce a plug of a sample into the mobile phase without having to stop the mobile phase flow,& without introducing air into the system.
The mixture of components is carried in a narrow band to the top of the column.Some compounds in the sample mixture will have greater preference for stationary phase than the mobile phase and will be retained in the column longer.
POINT TO REMRMBER : "Like attracts Like" i.e if the column is non-polar the compound to elute first will be the most polar one.
Those components that are not retained as strongly as are carried by the mobile phase down the column.
The detector is then used to respond to a physico-chemical property of analyte,this response is digitally amplified and sent to a data system where it is recorded as a 'chromatogram'.
What else can I help you with?
What type of species can be separated by HPLC but not by gas liquid chromatography?
mixture of enantiomers can be separated by HPLC
Can melamine detect by HPLC?
Yes, melamine can be detected by HPLC (High Performance Liquid Chromatography). HPLC is a common analytical technique used to separate and quantify compounds in a mixture, including melamine. Detection methods such as UV-Vis spectroscopy or mass spectrometry can be used in conjunction with HPLC to identify and quantify melamine accurately.
Why volatile compounds are analysed in gc but not in hplc?
GC can give very resolved sharp peaks with short run time compared to hplc. additionally, there is less compatibility issue in setting an MS up to a GC than HPLC
Which are the top commercial uses for HPLC?
HPLC is used to analyze, identify, purify and quantify organic compounds.
Why ordinary liquid pump cannot be used in hplc system?
An ordinary liquid pump cannot be used in HPLC systems because HPLC requires pumps that can generate high pressure to push solvents through the column at a constant and precise flow rate. Ordinary pumps do not provide the level of pressure and flow rate control needed for HPLC separations, which can negatively impact the accuracy and reproducibility of chromatographic results. HPLC pumps are specifically designed to handle the high pressures and flow rates required for efficient separations.
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).
Retention time calculation for hplc?
why RT was shifting & how to RT calculation in HPLC
What type of species can be separated by HPLC but not by gas liquid chromatography?
mixture of enantiomers can be separated by HPLC
What is rs-hplc?
"RS-HPLC method" means "Related Substance HPLC Method".
Where can one purchase HPLC detectors?
You can purchase used HPLC detectors and other equipment from the usedhplc website or from the ebay bidding website. Alternatively you can buy HPLC detectors from the equipnet website.
What is RRT and RRF in hplc?
In HPLC RRT means Relative Retention Time and RRF is Relative Response Factor
What is dead volume HPLC?
The dead volume in HPLC is 137.45. The dead volume in science is used in retention measurements and also in thermodynamic studies and the abbreviation HPLC stands for High Pressure Liquid Chromatography.
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 melamine detect by HPLC?
Yes, melamine can be detected by HPLC (High Performance Liquid Chromatography). HPLC is a common analytical technique used to separate and quantify compounds in a mixture, including melamine. Detection methods such as UV-Vis spectroscopy or mass spectrometry can be used in conjunction with HPLC to identify and quantify melamine accurately.
What is an HPLC column?
HPLC Column is one type of tube containing a stationary phase react with mobile phase to detect peak
Why volatile compounds are analysed in gc but not in hplc?
GC can give very resolved sharp peaks with short run time compared to hplc. additionally, there is less compatibility issue in setting an MS up to a GC than HPLC
Which one is advanced gc or hplc?
HPLC (High-performance liquid chromatography) is generally considered more advanced than GC (Gas chromatography) due to its broader application range, higher sensitivity, and ability to separate a wider range of compounds. HPLC is often preferred for analyzing complex mixtures and compounds that are not volatile.
