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Can you use a C18 column for HPLC with fluorescence detector and methanol as the mobile phase?
Wiki User
∙ 15y ago
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.
Wiki User
∙ 15y ago
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Why you note optical density of protein in chromatography technique?
Chromatography is a group of techniques to divide components of a mixtures basically on the ground of their physical dimension, even if elected types of chromatography exists where separation happens for example on the ground of electron affinity. In general terms the mixture is dissolved in a fluid called the mobile phase, which carries it through a structure holding another material called the stationary phase. The various constituents of the mixture travel at different speeds in the stationary phase, causing them to separate. Liquid phase chromatography is the most used type of chromatography, where the mobile phase is a liquid and the stationary phase a material composed of very small particles strictly packed one with the other. Also gas chromatography, where the mobile phase is a gas, is largely used. The name chromatography derives from the fact that the very first version of this separation techniques used different colors added to the mobile phase in different moments to visually distinguish the components coming out from the chromatographic column in different moments, but this technique is no more used, substituted by more sophisticate ways to quantify components that comes out in different instants from the chromatographic column. Optical detection is frequently used, generally illuminating the flow out of the column with UV light and observing fluorescence lines. The fluorescence intensity is proportional for diluted solutions to the concentration of the substance presenting the observed optical transition. Since many proteins are characterized by well identified fluorescence lines this detection method can be used for proteins quantification.
Basic parts of hplc?
(1) Solvent reservoirs, (2) Solvent degasser, (3) Gradient valve, (4) Mixing vessel for delivery of the mobile phase, (5) High-pressure pump, (6) Switching valve in "inject position", (6') Switching valve in "load position", (7) Sample injection loop, (8) Pre-column (guard column), (9) Analytical column, (10) Detector (i.e. IR, UV), (11) Data acquisition, (12) Waste or fraction collector. Please seehttp://en.wikipedia.org/wiki/File:HPLC_apparatus.svg
What is the importance of a mixed elution solvent in column chromatography?
In column chromatography, the stationary phase, a solid adsorbent, is placed in a vertical glass (usually) column and the mobile phase, a liquid, is added to the top and flows down through the column (by either gravity or external pressure). Column chromatography is generally used as a purification technique: it isolates desired compounds from a mixture.
Is IPA can be used for mobile phase in column chromatography?
Yes why not! i didn't try it but u can try it to observe results. it is a viscous solvent may be fraction come out slowly.
What is dead volum in 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.
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 is mean by mobile sniffer?
it is a mobile phone detector
Will a metal detector detect a mobile phone in grass?
Yes the metal detector would but it is a large object and you would see it also.
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 mobile phase?
The mobile phase as indicated is the moving phase. Either the mobile or stationary phase is polar and the other is Non-polar. A common polar phase is Methanol, and non-polar is hexane
Do people really use a portable gas detector?
Yes people use a portable gas detector. This enables them to move their location and detect gas there. This can be better if you are more mobile and going to different locations.
What is an HPLC column?
HPLC Column is one type of tube containing a stationary phase react with mobile phase to detect peak
What will be the mobile phase for Thin Layer Chromatography of Schiff base?
Schiff bases are imines formed by the condensation of aldehydes or ketones with primary amines. The mobile phase for this could be ethyl acetate in hexane or chloroform in methanol.
Stability indicating method for febuxostat tabs by hplc?
Methods The determination of febuxostat and the separation of its related substances was performed on a C18 column( 200 mm ×4.6 mm, 5 μm ) . The mobile phase was methanol-acetonitrile-0.05%(w)phosphoric acid (V:V:V=24:46:30) . The flow rate was 1.0 mL·min-1. Ultraviolet absorption detector was set at 315 nm and column temperature at 35 ℃. Results The linear range of febuxostat was between15.7 and 94.3 mg·L-1 ( r = 0. 999 8) . The average recovery was 100.5% with RSD of 1.0%.The related substances of febuxostat could be completely separated from febuxostat. The limit of detection (LOD) was 0.5 ng.
Why you note optical density of protein in chromatography technique?
Chromatography is a group of techniques to divide components of a mixtures basically on the ground of their physical dimension, even if elected types of chromatography exists where separation happens for example on the ground of electron affinity. In general terms the mixture is dissolved in a fluid called the mobile phase, which carries it through a structure holding another material called the stationary phase. The various constituents of the mixture travel at different speeds in the stationary phase, causing them to separate. Liquid phase chromatography is the most used type of chromatography, where the mobile phase is a liquid and the stationary phase a material composed of very small particles strictly packed one with the other. Also gas chromatography, where the mobile phase is a gas, is largely used. The name chromatography derives from the fact that the very first version of this separation techniques used different colors added to the mobile phase in different moments to visually distinguish the components coming out from the chromatographic column in different moments, but this technique is no more used, substituted by more sophisticate ways to quantify components that comes out in different instants from the chromatographic column. Optical detection is frequently used, generally illuminating the flow out of the column with UV light and observing fluorescence lines. The fluorescence intensity is proportional for diluted solutions to the concentration of the substance presenting the observed optical transition. Since many proteins are characterized by well identified fluorescence lines this detection method can be used for proteins quantification.
What is void volume in hplc?
Total volume of mobile phase in a fully wet packed column- the space between the particles of the stationary phase plus the volume within the particles
Basic parts of hplc?
(1) Solvent reservoirs, (2) Solvent degasser, (3) Gradient valve, (4) Mixing vessel for delivery of the mobile phase, (5) High-pressure pump, (6) Switching valve in "inject position", (6') Switching valve in "load position", (7) Sample injection loop, (8) Pre-column (guard column), (9) Analytical column, (10) Detector (i.e. IR, UV), (11) Data acquisition, (12) Waste or fraction collector. Please seehttp://en.wikipedia.org/wiki/File:HPLC_apparatus.svg
