The major difference between HPLC and LC-MS is that HPLC uses phosphate buffer in the mobile phase, whereas LC-MS uses formic acid to avoid MS contamination. MS is 1000X more sensitive than the UV or PDA detector used in HPLC.
Chromatography is a technique of separation in chemistry, where analytes are separated using the mobile phase and stationary phase. There are different types of chromatography such as paper chromatography, column chromatography, thin-layer chromatography (TLC), and gas chromatography, adsorption chromatography, partition chromatography, and affinity, etc. each type works on the different principle.
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HPLC is a separation method, whereas MS is used to evaluate the mass-to-charge ratio of charged particles, in which the chromatographic conditions of both techniques are about the same. The technique of combining HPLC and MS is known as LC-MS. Combining the two techniques reduces error and improves accuracy. Liquid chromatography-mass spectrometry is valuable in applications that contain a large number of compounds.
What is HPLC?
High-performance liquid chromatography is a popular separation process used in analytical chemistry to identify and quantify each compound in a sample mixture. Since it relied on pumps to flow a pressurized liquid solvent containing the sample combination through a column filled with a solid adsorbent (Generally, silica) material, this technique was previously known as high-pressure liquid chromatography.
The sample is injected through the injector, and the mobile phase flows it through the column. The solid adsorbent material interacts differently with each analyte in the sample mixture, resulting in different flow rates for various analytes. The separation and retention time depend on the affinity of the analyte towards the stationary phase.
What is LC-MS?
LC-MS is a sample analytical technique that combines liquid chromatography (LC) and mass spectrometry (MS). Mass spectrometry is used to determine the mass-to-charge ratio of charged particles, while liquid chromatography is a separation technique.
In comparison to HPLC, LCMS is a popular analytical technique with high sensitivity, accuracy, and specificity. As a result, it has a wide range of applications, including pharmaceutical, environmental, research, and food analysis, etc. In the presence of complex sample mixtures, LCMS is primarily used to separate, detect, identify, and quantify the biochemical properties of a specific sample.
Difference between HPLC and LCMS:
- HPLC is a separation technique that employs mainly to separate, identify and determine each component in the sample of a complex mixture, while LC-MS is a hybrid system in which a UV or PDA detector of HPLC is replaced with a mass spectrometer.
- The accuracy and sensitivity of HPLC results are less than LCMS, while LCMS results are highly accurate and sensitive than HPLC.
- HPLC is a time-consuming method than the LCMS, while LCMS has a faster analysis than HPLC.
- HPLC is a liquid chromatography technique only, while the LCMS is a combination of liquid chromatography and the mass spectroscopy method.
- HPLC can be considered as a component of LCMS, while LCMS can be considered as a component of HPLC.
- The HPLC analysis is less specific than the LCMS, while the LCMS analysis is more specific than the HPLC.
- Using an HPLC the biomolecules, organic molecules, polymers, and ions can be analyzed, while using an LCMS the biomolecules and organic molecules can be analyzed.
- An instrument of high-performance liquid chromatography does not have an ion source, while in the instrument of liquid chromatography-mass spectrometry the ion source is present.
HPLC vs LC-MS
Which is better HPLC or LCMS?
LCMS is more precise, specific, accurate, and sensitive than HPLC. The HPLC is part of LC-MS that separates solutes within a sample, and the mass spectrometer offers mass to charge ratio data that can help determine the compound's structural identification.
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