Introduction of Chromatogram and Peak Parameters of High Performance Liquid Chromatograph

Abstract: High performance liquid chromatography only requires that the sample can be made into a solution, which is not limited by the volatility of the sample. The mobile phase can be selected from a wide range. There are many types of stationary phases, so it can separate thermally unstable and nonvolatile, dissociative And non-dissociative substances in various molecular weight ranges.

1. Chromatogram (chromatogram)-the sample flows through the chromatographic column and detector, the resulting signal-time curve, also known as chromatogram elution profile (elutionprofile).

2. Baseline-The mobile phase is flushed. After the column and mobile phase reach equilibrium, the detector measures the outflow curve for a period of time. Generally it should be parallel to the time axis.

3. Noise-the fluctuation of the baseline signal. Usually caused by poor power contact or transient overload, unstable detector, mobile phase containing bubbles or chromatographic column contamination.

4. The drift baseline changes slowly with time. Mainly due to the unstable operating conditions such as voltage, temperature, mobile phase and flow rate, the contaminants or stationary phase in the column will continue to elute and drift.

5. Chromatographic peak (peak)-the curve produced by the corresponding continuous signal when the component flows through the detector. The protruding part on the outflow curve. Normal chromatographic peaks approximate a symmetric normal distribution curve (Gaussian Gauss curve). There are two types of asymmetric chromatographic peaks: leading peak and tailing peak. The former is rare.

6. Tailing factor (T)-T = B / A, used to measure the symmetry of chromatographic peaks. Also known as symmetry factor (symmetryfactor) or asymmetry factor (asymmetryfactor) "Chinese Pharmacopoeia" stipulates that T should be 0.95 ~ 1.05. T1.05 is the tailing peak.

7. The bottom of the peak-the distance from the start point to the end point of the peak on the baseline.

8. Peak height (Peakheight, h)-the distance from the highest point of the peak to the bottom of the peak.

9, peak width (peakwidth, W)-the distance between the two tangents made at the inflection points on both sides of the peak and the two intersections of the baseline. W = 4σ.

10. Half-width (peakwidthathalf-height, Wh / 2)-the peak width at half of the peak height. Wh / 2 = 2.355σ.

11. Standard deviation (σ)-half of the peak width when the normal distribution curve x = ± 1 (inflection point). The inflection point of the normal peak width is 0.607 times the peak height. The size of the standard deviation indicates the degree of dispersion of the components during the flow out of the column. σ is small, the degree of dispersion is small, the pole concentration is high, the peak shape is thin, and the column efficiency is high; otherwise, the σ is large, the peak shape is fat, and the column efficiency is low.

12. Peak area (peakarea, A)-the area enclosed by the peak and the bottom of the peak. A = × σ × h = 2.507σh = 1.064Wh / 2h.


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