Full analysis of HPLC principles: analysis methods, benefits, operating procedures and detection applications
The HPLC principle may seem complicated, but the core lies in "separate first, then detect". This article will start with 3 major analysis methods, 6 operating steps, and 2 common questions to help you understand how HPLC separates samples, interprets spectra, and masters the precautions when entrusting professional laboratories to perform testing.
Table of Contents
(1) What is HPLC? Comparison of the differences between HPLC, FPLC and GC
(2) HPLC principle analysis: understand it from the analysis mechanism to common methods at once
(3) What are the uses of HPLC? HPLC Qualitative and Application Analysis
(4) What are the advantages of HPLC? Why has it become a mainstream analytics technology?
(1) HPLC instrument composition and functional architecture
(2) HPLC operating principle and substance separation mechanism description
(3) HPLC standard operating steps and process description
(1) Why are the HPLC measurement times different?
(2) Why is the HPLC pressure too high?
1. Full analysis of HPLC principles: understanding the separation mechanism, operating procedures and application advantages at once
HPLC is a common high-performance liquid chromatography technology. Methods such as reversed-phase chromatography, normal-phase chromatography, ion exchange chromatography or size exclusion chromatography can be selected according to the characteristics of the sample. After the separation is completed, it is then used with UV-Vis, FLD, LC-MS and other detectors for qualitative and quantitative analysis.
HPLC can separate complex samples by adjusting mobile phase conditions through isocratic or gradient elution, so it is widely used in food inspection, pharmaceutical analysis, environmental monitoring and biotech medical research. Because of its high sensitivity, high resolution and good reproducibility, HPLC has become a commonly used mainstream analytical technology in laboratories.
(1) What is HPLC? Comparison of differences between HPLC, FPLC and GC
HPLC in Chinese is High Performance Liquid Chromatography. It is an analytical technology that uses high-pressure liquid as the mobile phase to push samples through a chromatography column. The principle is to use the force difference between different components in the sample and the stationary phase and mobile phase to make each component pass through the column at different speeds, thereby achieving the purpose of separation, qualitative and quantitative analysis.
From the actual analysis process, the core logic of HPLC is "separate first, then detect". When a sample contains multiple components, if detected directly, the signals are likely to interfere with each other, thus affecting the interpretation results. HPLC first separates different compounds according to their characteristics through a liquid phase system, and then uses a detector to read the signals of each component, thus improving the accuracy and stability of analysis.
Since HPLC does not require vaporization of the sample, it is suitable for analyzing samples that are not resistant to high temperatures, are not volatile, have complex composition, or are soluble in liquids. It is commonly used in the fields of medicine, food, environment, biotechnology, and chemical analysis.
In addition to HPLC, laboratories often choose FPLC (fast protein liquid chromatography) or GC (gas chromatography) depending on the purpose of analysis. Although these three techniques are all chromatography methods, their mobile phases, applicable samples, and main uses are different. The following is a one-time explanation for you:
