Some Basic Concepts In The Field Of HPLC Columns Chromatography
Chromatography, also known as chromatography or chromatography, is a physicochemical analysis method that uses the difference between the forces (distribution, adsorption, ion exchange, etc.) between different solutes (samples) and the stationary phase and mobile phase. When the two phases move relatively, each solute is equilibrated multiple times between the two phases, so that the solute is separated from each other. The following related concepts are commonly found in the use of HPLC columns.
Chromatogram related concepts
Chromatogram: A plot of the response signal generated by the HPLC column effluent as it passes through the detector system versus time or carrier gas volume.
Chromatographic peak: A differential curve of the response signal produced by the column as it exits the detector system.
Baseline: The line connecting the start and end of the peak.
Peak height: The distance from the peak maximum to the peak baseline.
Peak Width: The distance between the tangent at the inflection point on both sides of the peak and the point at which the peak baseline intersects.
Half-width: The line passing through the midpoint of the peak height equal to the bottom of the peak, the distance between the line and the two sides of the peak.
Peak area: The area between the peak and the peak baseline.
Related concepts of chromatographic processing
Data acquisition: During the process of collecting data, the signal output by the analytical instrument is converted from an analog signal to a digital signal in the collector. The digital signal is sent to the N2010 chromatography station and saved in the signal data file.
Integration: Integration is to determine the peak from the signal curve and calculate its size. Points are essential for quantitative calculations. When integrating the N2010 Chromatography Workstation, first identify the start and end times of each peak, and mark the points with the “|” symbol, find the vertices of these peaks, determine the retention time, establish a baseline, and calculate the peak area, peak height, and peak width. These processes are controlled by integral parameters and manual integration event tables.
Quantitation: Use peak area or peak height to determine the concentration of a compound in a sample, including the following: clarify and identify the compound you are analyzing; establish a method for analyzing a sample containing the compound; analyze one or more of the concentrations of the known compound Standard samples to obtain the response at this concentration, and calculate the correction factor; analyze the sample of the compound at an unknown concentration to obtain the response of the unknown concentration; compare the sample of unknown concentration with the standard sample, and use the correction factor of the standard sample To determine the concentration of a compound in an unknown sample.
Calibration: Calibration is the process of determining the correction factor for the absolute component concentration by injecting the specified prepared standard sample.
The number of repetitions: The number of parallel injections of standard samples of the same concentration.
Calibration points: Consists of a calibration point that calibrates different sample concentrations.
Standard sample: Also called a calibration sample or standard mixture, is a sample containing a known amount of compound for quantification. Standard samples are available from national standard reagent suppliers.
Standard curve: A graphical representation of the number of compounds and the cause data obtained from one or more standard samples.
Report: The report contains information on the quality and quantity of the sample being analyzed. Reports can be printed directly or displayed on the screen, and the report can include details of the peaks measured during the run and the resulting spectra.