1923–1970

Partition and Lipid Chromatography Evolution

Partition-driven thinking and two-phase liquid systems established the core framework for systematic biomolecule separations. Paper-based partition methods and colorimetric detection enabled quantitative analyses of amino acids, proteins, and vitamins. Thin-layer and column techniques expanded analytical capabilities for complex lipid mixtures and sugars. Automated instrumentation and the retention index system improved reproducibility and enabled cross-laboratory library-based identification.

Period 1923-1947: Researchers separated biomolecules and vitamins using partition, paper, and colorimetric methods, supported by clinical workflows and new indicators[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15]. Foundational theoretical frameworks and liquid-liquid partition techniques guided subsequent method design and solvent-system development[16][2][8]. Period 1948-1970: Thin-layer and gas chromatography isolated lipids, sugars, and amino acids using automated recorders, papers, and DEAE-cellulose columns[17][18][19][20][21][22][23][24][25][26][27][28][29][30][31]. Retention indices, automated recording, two-dimensional thin-layer methods, colorimetric sugar assays, and phosphorus detection shaped later analytical chemistry[32][24][33][34][35].

1971–1995

Advancements in Reversed-Phase Liquid Chromatography

High-performance liquid chromatography matured through advances in column packing, particle size, and chemically bonded stationary phases that enabled faster analyses. Systematic evaluations of bond chemistry and porosity-focused media guided predictable retention patterns for complex biochemical separations. Gradient-driven control of retention and selectivity subsequently emerged as the prevailing paradigm for small molecules and peptides. Stationary-phase engineering expanded to cyclodextrin architectures while fundamental liquid chromatography behavior provided practical, predictive insights into method robustness.

Period 1971-1977: Engineered small-particle silica packings and chemically bonded stationary phases advanced high-speed reversed-phase liquid chromatography for biomedical and metabolic analyses[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]. Influential chromosome banding, sister chromatid staining, and Concanavalin A affinity methods shaped modern cytogenetics and protein separation workflows[18][19][20][21]. Period 1978-1995: Gradient elution, stationary phase engineering, and fundamental chromatographic studies optimized reversed-phase separations for peptides, biomolecules, and trace analytes[22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37]. These methodological advances enabled rapid preparative techniques, ultra-sensitive amino acid assays, peptide hydrophilicity mapping, DNA GC content quantification, and standard volatile analyses[38][39][40][41].

1996–2009

Multidimensional Hyphenated Chromatography and Mass Spectrometry

Multidimensional hyphenated chromatography became the dominant paradigm for analyzing complex mixtures. Innovations in stationary phase design and monolithic packings significantly improved separation efficiency. Advanced sample preparation techniques integrated seamlessly with mass spectrometry detection. This established robust workflows for high-precision trace analysis across diverse matrices.

Period 1996-2002: Hyphenated two-dimensional chromatography and mass spectrometry coupled advanced stationary phases and integrated sample preparation to resolve complex biological and environmental matrices[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]. Foundational separation principles, metal-affinity phosphoproteomics, and matrix-effect evaluations shaped subsequent quantitative workflows and natural-product analytics[18][19][20][21][10]. Period 2003-2009: Researchers coupled dispersive extraction and orthogonal two-dimensional liquid chromatography with mass spectrometry to achieve reliable multi-residue trace quantification in complex matrices[22][23][24][25][26][27][28][29][30][31][32][33][34][35][36]. Foundational works optimized GC-MS spectral libraries, multi-stage LC-MS fragmentation annotation, nonlinear preparative scale-up, and counter-current solvent selection to shape modern analytical workflows[37][38][39][40].

2010–2023

High-Throughput and Cannabinoid Chromatographic Profiling

Automated online sample preparation and advanced sorbent materials matured within liquid chromatography tandem mass spectrometry workflows for multi analyte screening. Hydrophilic interaction liquid chromatography and automated feature deconvolution enhanced complex metabolomic and environmental analyses. Integrated chromatographic platforms paired with high resolution mass spectrometry enabled untargeted profiling and targeted residue analysis in botanical and food products. Novel extraction strategies and multi dimensional separation approaches delivered superior resolution for regulatory compliance and quality assessment.

Period 2010-2016: Automated online SPE, MOFs, and hybrid TOF systems enabled high-throughput LC-MS/MS trace screening across matrices[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]. CAMERA[18], HILIC[19], GC-MS continuity[20], TiO2 enrichment[21], and bisphenol assays[22] shaped future data analysis, metabolomics, and environmental monitoring. Period 2017-2023: Coupled chromatographic platforms and advanced preconcentration strategies enabled high-resolution metabolome profiling, multidimensional separations, and cannabinoid standardization across complex matrices[23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40]. Foundational GC syntheses, modern TLC appraisals, hemp extraction workflows, and LC-HRMS profiling established enduring standards for accuracy, validation, and multi-residue analysis[41][42][32][43].

Partition and Lipid Chromatography Evolution era

F. Smith and 3 others

F. Smith[1] conducted research while affiliated with the University of Minnesota[5] and the University of California, Los Angeles[6]. Their work advanced colorimetric detection for quantitative analyses through Colorimetric Method for Determination of Sugars and Related Substances[10], which expanded analytical capabilities for complex sugars. P. A. Rebers[2] performed investigations at Rutgers, The State University of New Jersey[7] and the National Animal Disease Center[8]. Through Colorimetric Method for Determination of Sugars and Related Substances[10], they contributed to the colorimetric detection methods that enabled systematic biomolecule and sugar measurements. J. K. Hamilton[3] pursued studies at the University of Minnesota[5]. They advanced the expansion of analytical capabilities for complex sugar mixtures via Colorimetric Method for Determination of Sugars and Related Substances[10], supporting quantitative biomolecule analysis. K. A. Gilles[4] was based at North Dakota State University[9] and the University of Minnesota[5]. They helped establish quantitative colorimetric analyses for complex mixtures through Colorimetric Method for Determination of Sugars and Related Substances[10].

Advancements in Reversed-Phase Liquid Chromatography era

W. Clark Still and 2 others

W. Clark Still[11] conducted research while based at Columbia University[14]. Through the publication of Rapid chromatographic technique for preparative separations with moderate resolution[19], this work contributed to advancements in reversed-phase liquid chromatography by introducing faster analytical and preparative capabilities. Michaël Kahn[12] developed further insights while affiliated with the University of Illinois Chicago[15] and the University of Washington[16]. The study Rapid chromatographic technique for preparative separations with moderate resolution[19] advanced the era's main theme by refining techniques for rapid separation processes. Abhijit Mitra[13] pursued investigations at the University of California, Davis[17] and the CSIR National Physical Laboratory of India[18]. The findings presented in Rapid chromatographic technique for preparative separations with moderate resolution[19] contributed to the methodological refinements characteristic of this era in liquid chromatography.

Multidimensional Hyphenated Chromatography and Mass Spectrometry era

Steven J. Lehotay and 3 others

Steven J. Lehotay[20] conducted research while based at the Eastern Regional Research Center[24] and the Agricultural Research Service[25]. He advanced high-precision trace analysis across diverse matrices through his foundational work in Fast and Easy Multiresidue Method Employing Acetonitrile Extraction/Partitioning and “Dispersive Solid-Phase Extraction” for the Determination of Pesticide Residues in Produce[30] and further established robust analytical workflows via Validation of a Fast and Easy Method for the Determination of Residues from 229 Pesticides in Fruits and Vegetables Using Gas and Liquid Chromatography and Mass Spectrometric Detection[31]. Frank J Schenck[21] performed investigations at the United States Food and Drug Administration[26] and the Center for Food Safety and Applied Nutrition[27]. He contributed to the dominant paradigm of complex mixture analysis by utilizing Fast and Easy Multiresidue Method Employing Acetonitrile Extraction/Partitioning and “Dispersive Solid-Phase Extraction” for the Determination of Pesticide Residues in Produce[30] to streamline sample preparation workflows for mass spectrometry detection. Michelangelo Anastassiades[22] contributed to this field while working at the Eastern Regional Research Center[24] and the Chemisches und Veterinäruntersuchungsamt Stuttgart[28]. He shaped advanced sample preparation techniques through the development reported in Fast and Easy Multiresidue Method Employing Acetonitrile Extraction/Partitioning and “Dispersive Solid-Phase Extraction” for the Determination of Pesticide Residues in Produce[30], supporting robust trace analysis workflows. Darinka Štajnbaher[23] carried out scientific studies at the Institute for Environmental Protection and Sensors[29]. She aided the integration of advanced sample preparation with mass spectrometry detection by co-authoring Fast and Easy Multiresidue Method Employing Acetonitrile Extraction/Partitioning and “Dispersive Solid-Phase Extraction” for the Determination of Pesticide Residues in Produce[30].

High-Throughput and Cannabinoid Chromatographic Profiling era

H. Brzuszkiewicz and 2 others

H. Brzuszkiewicz[32] contributed to high-throughput and cannabinoid chromatographic profiling through foundational separation research. Their work on the paper thin layer chromatography[35] advanced multi analyte screening capabilities by refining separation techniques applicable to complex botanical and quality assessment workflows. H. Kraczkowski[33] also contributed to high-throughput and cannabinoid chromatographic profiling within the era's analytical framework. Their involvement with the paper thin layer chromatography[35] enhanced complex metabolomic and environmental analyses through improved resolution strategies. J OPIENSKA BLAUTH[34] further supported high-throughput and cannabinoid chromatographic profiling by participating in vital separation studies. Their contribution through the paper thin layer chromatography[35] advanced integrated chromatographic platforms paired with high resolution mass spectrometry for targeted residue analysis.