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What is difference between an HPLC UV detector and a fluorescence detector?
HPLC UV detectors measure absorbance of UV light at a specific wavelength, while fluorescence detectors measure the emission of light at a longer wavelength after excitation with UV light. Fluorescence detectors are more sensitive and selective than UV detectors, but may require additional steps such as derivatization for certain 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.
How does an RID detector work for HPLC?
An RID (Refractive Index Detector) for HPLC works by measuring changes in refractive index caused by the presence of analytes eluting from the column. As analytes pass through the detector cell, they displace the mobile phase, causing changes in refractive index that are detected and converted into a signal. The signal is then plotted against retention time to create a chromatogram that can be used to identify and quantify analytes in the sample.
Definition of standard in HPLC?
In HPLC, a standard is a known compound with a defined chemical structure and purity used for comparison and identification purposes. Standards are essential for calibrating instruments, determining retention times, and quantifying unknown compounds in samples during analysis.
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.
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 between an HPLC UV detector and a fluorescence detector?
HPLC UV detectors measure absorbance of UV light at a specific wavelength, while fluorescence detectors measure the emission of light at a longer wavelength after excitation with UV light. Fluorescence detectors are more sensitive and selective than UV detectors, but may require additional steps such as derivatization for certain compounds.
Why is Erbium perchlorate is used in calibration of HPLC?
Erbium has a strong absorption in uv and visible range, It is used in HPlc calibration for the wavelength accuracy verification of the PDA detector.
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 difference between pda and uv detector in hplc?
PDA - Photo diode array UV- Ultra violet with use of PDA detector, we can measure the area or height of particular peak at different wavelengths ranging from 200 to 800 nm by injecting the solution at once. Where as in uv detector we can measure the area or height of the peak only at two different wavelengths. But that wavelengths also to be selected before injecting the solution.
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 HPLC Calibration parameters and elaborate it?
1. Flow rate 2. Temp. of column 3. Detector function 4. Resolution
How does an RID detector work for HPLC?
An RID (Refractive Index Detector) for HPLC works by measuring changes in refractive index caused by the presence of analytes eluting from the column. As analytes pass through the detector cell, they displace the mobile phase, causing changes in refractive index that are detected and converted into a signal. The signal is then plotted against retention time to create a chromatogram that can be used to identify and quantify analytes in the sample.
Definition of standard in HPLC?
In HPLC, a standard is a known compound with a defined chemical structure and purity used for comparison and identification purposes. Standards are essential for calibrating instruments, determining retention times, and quantifying unknown compounds in samples during analysis.
What is post run in HPLC?
Post run in HPLC refers to the time after the completion of a chromatographic analysis where the system continues running to ensure that any remaining compounds are fully flushed out of the column and detector to prevent contamination and achieve a clean baseline for subsequent runs. It is an important step to maintain the integrity and performance of the HPLC system.
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 difference between dad and uv detector in hplc?
Liquid chromatography (LC) encompasses all chromatographic techniques using liquid mobile phase, including planar chromatography (paper chromatography and thin-layer chromatography) and column chromatography (classical column chromatography, and high-performance liquid chromatography on packed and capillary columns). The term liquid chromatography is nowadays often used as a sinonim for high performance liquid chromatography (HPLC) and ultra-high performance liquid chromatography (UHPLC).
