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High-Performance Liquid Chromatography: History
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Contributor: Yu Peng

High-performance liquid chromatography (HPLC) is a separation technique in analytical chemistry in which a liquid mobile phase is forced under elevated pressure through a column packed with a stationary phase, effecting the differential migration of dissolved sample components according to their distribution between the two phases [1][2]. The technique employs a liquid mobile phase, typically a mixture of two or more solvents, delivered by a high-pressure pump at inlet pressures sufficient to drive flow through columns packed with fine solid particles of narrow size distribution [2][3]. Separation arises from the repeated equilibrium partitioning of analyte molecules between the stationary phase and the mobile phase as the mobile phase percolates through the column, causing components with different affinities for the stationary phase to elute at characteristic retention times. The instrumentation comprises a solvent delivery system, a sample injection valve, a chromatographic column containing the stationary phase, a detector, and a data acquisition unit [1]. HPLC encompasses multiple separation modes—including reversed-phase, normal-phase, ion-exchange, and size-exclusion chromatography—selected according to the physicochemical properties of the analytes and the composition of the stationary and mobile phases. The technique is defined by its capacity to resolve, identify, and quantify components within complex mixtures through pressure-driven liquid-phase chromatography [1][3].

  • Liquid mobile phase
  • high-pressure pumping
  • differential partitioning
  • column chromatography

References

  1. Luigi Mondello; Paola Dugo; Paola Donato; Miguel Herrero; Lidia Montero; Oliver J. Schmitz; Comprehensive two-dimensional liquid chromatography. Nat. Rev. Methods Prim. 2023, 3, 1-17, 10.1038/s43586-023-00269-0.
  2. Petar Žuvela; Magdalena Skoczylas; J. Jay Liu; Tomasz Ba̧cZek; Roman Kaliszan; Ming Wah Wong; Bogusław Buszewski; Column Characterization and Selection Systems in Reversed-Phase High-Performance Liquid Chromatography. Chem. Rev. 2019, 119, 3674-3729, 10.1021/acs.chemrev.8b00246.
  3. Matthew J. Sorensen; Brady G. Anderson; Robert T. Kennedy; Liquid chromatography above 20,000 PSI. TrAC Trends Anal. Chem. 2020, 124, 115810, 10.1016/j.trac.2020.115810.
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