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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.
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
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.
How do you determine lauric acid in coconut milk using hplc?
To determine lauric acid in coconut milk using HPLC, you would first extract the fatty acids from the coconut milk using a suitable solvent. Then, prepare the HPLC system with a column and mobile phase that can separate lauric acid from other compounds present. Inject the extracted sample into the HPLC system and use a suitable detector to quantify the amount of lauric acid present based on retention time and peak area.
What is the H.P.L.C. chromatography?
HPLC stands for high performance liquid chromatography. It is a liquid chromatography which involves the separation of the compounds on the basis of their polarity. It is used to analyze, identify, purify & quantify the compounds.
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 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 the difference between low pressure and high pressure gradient HPLC?
In a high pressure gradient pump, each different mobile phase is delivered by an individual pump head and then the mobile phases are mixed at the pump outlet. In a low pressure gradient pump, different mobile phases are mixed using a valve before entering the pump head. As a result of the fact that the low-pressure gradient design uses only one pump head, it is of lower cost. It can also use more types of mobile phase without significant increase of cost. Since solvent mixing point is much closer to the column head in the high-pressure gradient design, it provides a much faster gradient. This is measured using delay volume. The value can be 50-300 uL for high pressure gradient pump and can be 2 to 3 times larger for a low pressure gradient pump. A small delay volume is important when the analysis time is short or the flow rate is low. If the delay volume is too large, it become impossible to obtain reproducible gradient run since the planed composition cannot reach the column head before a run is finished.
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
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.
How do you determine lauric acid in coconut milk using hplc?
To determine lauric acid in coconut milk using HPLC, you would first extract the fatty acids from the coconut milk using a suitable solvent. Then, prepare the HPLC system with a column and mobile phase that can separate lauric acid from other compounds present. Inject the extracted sample into the HPLC system and use a suitable detector to quantify the amount of lauric acid present based on retention time and peak area.
What is the H.P.L.C. chromatography?
HPLC stands for high performance liquid chromatography. It is a liquid chromatography which involves the separation of the compounds on the basis of their polarity. It is used to analyze, identify, purify & quantify the compounds.
How to interpret a HPLC chromatogram effectively?
To interpret a HPLC chromatogram effectively, first identify the peaks representing different compounds. Then, analyze peak shape, height, and area to determine concentration and purity. Compare retention times to standards for identification. Consider factors like column efficiency and mobile phase composition. Finally, use software or calculations to quantify results accurately.
What is the use of gradient elution analysis?
Gradient elution analysis is used in chromatography to improve separation of complex mixtures by changing the composition or strength of the mobile phase over time. This technique enables better resolution of components that may have similar retention times in isocratic elution. Gradient elution is particularly useful for separating compounds with a wide range of polarities or concentrations.
How is protein purification achieved using HPLC techniques?
Protein purification using HPLC techniques involves separating proteins based on their size, charge, and hydrophobicity. The process involves passing a protein mixture through a column filled with a stationary phase that interacts with the proteins differently, allowing for their separation. The proteins are then eluted from the column using a solvent gradient, with each protein eluting at a different time based on its unique characteristics. This results in the isolation of pure proteins for further analysis or use.
How do you change from reversed phase chromatography to normal phase chromatography?
How do you change from reversed phase chromatography to normal phase chromatography? answer:Water -------> Ethanol ---------> Acetone -----> Ethyl acetate ------>Chloroform ------->HeptaneHow to Change from normal phase to reversed phase chromatography?Heptane ------->Chloroform -------> Ethyl acetate ---->Acetone --------->ethanol -------> WaterMohammad Abdel Qader (Mousa)Lab. SupervisorChemical , Biological and Drug Analysis CenterAn-Najah National University.Nablus Palestinezawatehm@gmail.com 1)To ues reverse phase chromatography solvents like:-Acetonitrile,Methanol in HPLC Grade 2) To use normal phase chromatography sovents like:-Iso propyl alcohol,n-Haxane HPLC Grade
How does chiral Hplc work?
As a very crude analogy, think of a playground slide with dents in it that are the size and shape of a left shoe print. If we put some shoes at the top and let them slide down, the left shoes will be more likely to get stuck in the dents than the right shoes (which don't fit as well and therefore slide over them more easily).
