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What is HPLC and how do you use it?
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.
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propyl paraben being a stable compound with reproducible chromatographic parameters used as a internal standard for the calibration of the HPLC systems
This is a difficult question to answer because there are so many uses for the HPLC. The HPLC is widely used to separate a complex mixture into individual compounds. Sc…ientists use standards in order quantify the amount of a particular compound they have by generating calibration curves. How all this is done... Choose an eluent that will separate your compounds - test out different eluents on a TLC plate to make sure that your compounds are separating. Set the temperature on the HPLC so that it is high enough to evaporate the eluent. Insert your sample into the injection port (typically around 20 uL). Wait for your peaks to all come out. There are different ways to miminimize the amount it takes to run samples, but this is unique to your experiment, and so you have to just try out different things.
To separate mixtures.
large columns, specifically for packing. * analytical columns, for quantitative analysis, usually accompanied by a UV-vis detector. * Narrow bore columns, for more sensiti…ve analysis * capillary columns, very this silica columns used almost exclusively with GC mass spectroscopy. * packed bed columns. with silica beads. and may have groups attached, e.g. C18.
Caffeine having different wavelents it having 2 maximas and 1 minima. 1st maxima is 205nm 2nd maxima is 273nm minima is 245nm and it is primary reference standa…rd and also suggested in pharmacopiea. -Rajesh,Orchid
UV cut off of acetone is 330 nm and it is easyly available,non expensive. you can use any other solvent which has UV cut off above 250.
it is retained to a lesser extent....has dual wavelength detection (245nm, 350nm) and gives a sharp and consistent peak.
Caffeineis dule absorbance,it gives multiwavelength responds like 205,245,273.
Analytical chemists and Polymer Chemists
caffein is a highly stable, this is show dual lamda max like 205 and 275 nm, it is not a degradable. this is cheep chemical.
it used a mobile phase to carry a sample through a tube containing a stationary phase which is a liquid coated on solid particles.
Caffeine responds in 3 wavelength regions i.e 196 - 208nm in which caffeine maxima at 203±2nm 240 - 248nm in which caffeine minima at 244±2nm 270 - 278nm in which caffeine m…axima at 273±2nm So it covers the operation range of HPLC UV detector to perform wavelength accuracy. Apart from this Caffeine elutes in approx 2 min when using any C18 column which helps in reducing run time. That s the caffeine is preferred than any other standards for HPLC calibration K S Badrinath
In the laboratory, acetonitrile is used as a medium-polarity solvent that is miscible with water and has a convenient liquid range. With a dipole moment of 3.84 D, acetonitril…e dissolves a wide range of ionic and nonpolar compounds and is useful as a mobile phase in HPLC and LCMS.
higly unretainable and has high absorption at 260 nm
High Performance Liquid Chromatography [HPLC] is used commercially in the production of foods like coffee, in order to determine their chemical compositions.
HPLC is normally used to detect Vitamin D levels in blood. Some people need this test to make sure they are getting enough vitamin d in their diets and such.