By Donald Parriott
This consultant for the practising chromatographer who desires a prepared resource of data on HPLC detection explores and compares latest detection platforms and detectors, outlines the typical difficulties linked to a given detector, and provides confirmed ways to averting such difficulties.
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Additional resources for A Practical Guide to HPLC Detection
A functional group that absorbs ultraviolet or visible light) cannot be detected. One example would be aliphatic hydrocarbons. A technique called inverse chromatography has been developed to get around the problem of detecting non-UV-absorbing analytes with an absorbance detector. A UV-absorbing component, such as terphthalic acid, is added to the mobile phase, which artificially elevates the baseline. When non-UV-absorbing components elute, they cause a negative peak because they absorb less than the mobile phase.
III. QUANTITATIVE MEASUREMENT The goal of quantitation is to accurately measure the concentration of one or more components in a solution or mixture. Usually, some measurement of the sample is made and compared to a known standard. In an analytical system 51 3 · Absorbance Detection that has no bias, the deviation of the measurements from the "true" value are totally random in nature. The precision of the data, as measured by the percent relative standard deviation, is a measure of how close the values obtained deviate from the "true" value.
3-6 Beam splitter. gradient elution or due to a sample solvent different than the mobile phase eluting at the void volume of the column, a shift in the baseline occurs. This is because some of the light entering the flow cell is bent so that it strikes the wall of the cell, and is either absorbed or is scattered at such an angle that it does not reach the photodiode at the cell exit. A change in the refractive index of the mobile phase changes the degree to which light is bent, and therefore how much light is lost.