{"id":16666,"date":"2023-06-15T10:30:52","date_gmt":"2023-06-15T08:30:52","guid":{"rendered":"https:\/\/www.rct-online.de\/magazin\/?p=16666"},"modified":"2026-07-07T15:39:10","modified_gmt":"2026-07-07T13:39:10","slug":"chromatography-the-history-of-separation-techniques","status":"publish","type":"post","link":"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/","title":{"rendered":"Chromatography: From Colour Writing to Modern Separation Science"},"content":{"rendered":"<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_73 ez-toc-wrap-center counter-hierarchy ez-toc-counter ez-toc-white ez-toc-container-direction\">\n<div class=\"ez-toc-title-container\">\n<p class=\"ez-toc-title\" style=\"cursor:inherit\">Table of contents<\/p>\n<span class=\"ez-toc-title-toggle\"><a href=\"#\" class=\"ez-toc-pull-right ez-toc-btn ez-toc-btn-xs ez-toc-btn-default ez-toc-toggle\" aria-label=\"Toggle Table of Content\"><span class=\"ez-toc-js-icon-con\"><span class=\"\"><span class=\"eztoc-hide\" style=\"display:none;\">Toggle<\/span><span class=\"ez-toc-icon-toggle-span\"><svg style=\"fill: #999;color:#999\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" class=\"list-377408\" width=\"20px\" height=\"20px\" viewBox=\"0 0 24 24\" fill=\"none\"><path d=\"M6 6H4v2h2V6zm14 0H8v2h12V6zM4 11h2v2H4v-2zm16 0H8v2h12v-2zM4 16h2v2H4v-2zm16 0H8v2h12v-2z\" fill=\"currentColor\"><\/path><\/svg><svg style=\"fill: #999;color:#999\" class=\"arrow-unsorted-368013\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"10px\" height=\"10px\" viewBox=\"0 0 24 24\" version=\"1.2\" baseProfile=\"tiny\"><path d=\"M18.2 9.3l-6.2-6.3-6.2 6.3c-.2.2-.3.4-.3.7s.1.5.3.7c.2.2.4.3.7.3h11c.3 0 .5-.1.7-.3.2-.2.3-.5.3-.7s-.1-.5-.3-.7zM5.8 14.7l6.2 6.3 6.2-6.3c.2-.2.3-.5.3-.7s-.1-.5-.3-.7c-.2-.2-.4-.3-.7-.3h-11c-.3 0-.5.1-.7.3-.2.2-.3.5-.3.7s.1.5.3.7z\"\/><\/svg><\/span><\/span><\/span><\/a><\/span><\/div>\n<nav><ul class='ez-toc-list ez-toc-list-level-1 ' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Ferdinand_Runges_%E2%80%9CChemical_Coats_of_Arms%E2%80%9D\" title=\"Ferdinand Runge\u2019s \u201cChemical Coats of Arms\u201d\">Ferdinand Runge\u2019s \u201cChemical Coats of Arms\u201d<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Spot_Tests_and_pH_Measurement\" title=\"Spot Tests and pH Measurement\">Spot Tests and pH Measurement<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Ring_Chromatography_and_the_Role_of_Capillary_Action\" title=\"Ring Chromatography and the Role of Capillary Action\">Ring Chromatography and the Role of Capillary Action<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#From_Colour_Script_to_the_Purification_of_Natural_Products_The_First_Chromatography_Column\" title=\"From Colour Script to the Purification of Natural Products: The First Chromatography Column\">From Colour Script to the Purification of Natural Products: The First Chromatography Column<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#The_Slow_Rise_of_Column_Chromatography\" title=\"The Slow Rise of Column Chromatography\">The Slow Rise of Column Chromatography<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-6\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Chromatography_Becomes_a_Standard_Across_Many_Fields\" title=\"Chromatography Becomes a Standard Across Many Fields\">Chromatography Becomes a Standard Across Many Fields<\/a><ul class='ez-toc-list-level-3' ><li class='ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-7\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Size-Exclusion_Chromatography\" title=\"Size-Exclusion Chromatography\">Size-Exclusion Chromatography<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-8\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#High-Performance_Liquid_Chromatography\" title=\"High-Performance Liquid Chromatography\">High-Performance Liquid Chromatography<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-9\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Thin-Layer_Chromatography\" title=\"Thin-Layer Chromatography\">Thin-Layer Chromatography<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-10\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Gas_Chromatography\" title=\"Gas Chromatography\">Gas Chromatography<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-11\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Ion-Exchange_Chromatography\" title=\"Ion-Exchange Chromatography\">Ion-Exchange Chromatography<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-3'><a class=\"ez-toc-link ez-toc-heading-12\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Affinity_Chromatography\" title=\"Affinity Chromatography\">Affinity Chromatography<\/a><\/li><\/ul><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-13\" href=\"https:\/\/www.rct-online.de\/magazin\/en\/chromatography-the-history-of-separation-techniques\/#Ferdinand_Runges_Legacy_Today\" title=\"Ferdinand Runge\u2019s Legacy Today\">Ferdinand Runge\u2019s Legacy Today<\/a><\/li><\/ul><\/nav><\/div>\n<p style=\"text-align: justify;\"><strong>Where the history of chromatography begins remains uncertain. Nature itself performs the process: a chemical mixture separates into its individual components as it passes through an environment made of different materials. Around 1900, the American geochemist David Talbot Day (1859 \u2013 1925) observed one such case. He found that the components of crude oil, as they seep through certain porous or fine-grained rocks such as limestone, can sort themselves into long-chain, cyclic and aromatic hydrocarbons. In doing so, they leave differently coloured bands in the rock.<\/strong><\/p>\n<h2><span class=\"ez-toc-section\" id=\"Ferdinand_Runges_%E2%80%9CChemical_Coats_of_Arms%E2%80%9D\"><\/span>Ferdinand Runge\u2019s \u201cChemical Coats of Arms\u201d<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p style=\"text-align: justify;\">Separating mixtures of dyes still serves as a form of entertainment today. Dab a blot of ink onto a piece of blotting or filter paper. Then add a few drops of diluted alcohol to the same spot. The paper absorbs the liquid, and the various dyes that make up the ink migrate outwards from the centre in rings. The result is a set of imaginative patterns. Similar images can be produced from plant extracts, using petroleum spirit as a \u201cdeveloping fluid\u201d on absorbent paper.<\/p>\n<p style=\"text-align: justify;\">One person recognised the potential of such images early on: the German chemist Friedlieb Ferdinand Runge (1794 \u2013 1867). He explained them as <em>\u201cbonds formed through chemical interaction\u201d<\/em>. In 1855, he described them in detail in his treatise <em>\u201cDer Bildungstrieb der Stoffe, veranschaulicht in selbst\u00e4ndig gewachsenen Bildern\u201d<\/em> (\u201cThe Formative Drive of Substances, Illustrated in Independently Grown Images\u201d).<\/p>\n<div class=\"box info  \"><div class=\"box-inner-block\"><i class=\"fa tie-shortcode-boxicon\"><\/i>\n\t\t\tFrom today\u2019s perspective, it is debatable whether Runge\u2019s grown images \u2014 which he also called \u201c<em>chemical coats of arms<\/em>\u201d \u2014 already count as chromatography, or whether they are better described as a kind of spot test.\n\t\t\t<\/div><\/div>\n<h3><span class=\"ez-toc-section\" id=\"Spot_Tests_and_pH_Measurement\"><\/span>Spot Tests and pH Measurement<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">In a spot test, a drop of the dissolved sample is brought together with a specific reagent on a piece of filter paper. A change in colour then indicates the presence of a particular substance.<\/p>\n<p style=\"text-align: justify;\">The familiar acid\u2013base test using pH paper is one such spot test for <a href=\"https:\/\/www.rct-online.de\/magazin\/en\/ph-measurement-in-the-laboratory\/\">pH measurement<\/a>. The paper consists of filter-paper strips impregnated with an acid\u2013base colour indicator. It fell to the Austrian chemist Fritz Feigl (1891 \u2013 1971) to develop this simple spot test into a rapid analytical method. It remains in regular use in many chemical laboratories today.<\/p>\n<figure id=\"attachment_8806\" aria-describedby=\"caption-attachment-8806\" style=\"width: 450px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-8806 size-full\" title=\"Measuring the pH Value Using Impregnated pH Paper\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/messung-ph-werts.jpg\" alt=\"Measuring the pH value using impregnated pH paper\" width=\"450\" height=\"298\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/messung-ph-werts.jpg 450w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/messung-ph-werts-300x199.jpg 300w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/messung-ph-werts-310x205.jpg 310w\" sizes=\"(max-width: 450px) 100vw, 450px\" \/><figcaption id=\"caption-attachment-8806\" class=\"wp-caption-text\"><center>Measuring the pH Value Using Impregnated pH Paper<\/center><\/figcaption><\/figure>\n<h3><span class=\"ez-toc-section\" id=\"Ring_Chromatography_and_the_Role_of_Capillary_Action\"><\/span>Ring Chromatography and the Role of Capillary Action<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">Runge developed his \u201cchemical coats of arms\u201d on filter paper from mixtures of dyes, using various solvents. He watched how the solvents influenced the resulting colour patterns. He also let chemical reagents act on substances in the same way. By comparing these results with unknown substances, he could already draw conclusions about their composition. Runge\u2019s \u201cchemical coats of arms\u201d can therefore be seen as a forerunner of ring chromatography. This variant of paper chromatography is now rarely used.<\/p>\n<p style=\"text-align: justify;\">Among the first to study the mechanism behind this separation effect were two scientists working in 1861. The German-Swiss chemist and physicist Christian Friedrich Sch\u00f6nbein (1791 \u2013 1868) worked alongside his much younger colleague, the Swiss chemist Christoph Friedrich Goppelsr\u00f6der (1837 \u2013 1919). They observed how aqueous dye solutions diffused through paper and other absorbent materials. Differences in migration speed, they found, came down to capillary action.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"From_Colour_Script_to_the_Purification_of_Natural_Products_The_First_Chromatography_Column\"><\/span>From Colour Script to the Purification of Natural Products: The First Chromatography Column<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p style=\"text-align: justify;\">Chromatography as a practical method for separating substances only began with the work of Mikhail Semyonovich Tsvet (1872 \u2013 1919). This Russian botanist, born in Italy, described a method in 1901 for separating plant pigments from one another.<\/p>\n<div class=\"box note  \"><div class=\"box-inner-block\"><i class=\"fa tie-shortcode-boxicon\"><\/i>\n\t\t\tFor this, he used a vertical glass tube filled with finely ground calcium carbonate as the stationary, solid phase \u2013 the first <a href=\"https:\/\/www.rct-online.de\/en\/filtration\/liquid-chromatography-columns-for-hplc-and-mplc\">chromatography column<\/a>. He applied a leaf pigment extract to the top of the column and eluted it with a solvent mixture of petroleum ether and ethanol.\n\t\t\t<\/div><\/div>\n<p style=\"text-align: justify;\">The hydrophilic, green chlorophyll migrates more slowly through the carbonate layer than the less hydrophilic, yellow-orange carotenoids. As a result, the pigments accumulated at different points along the column. Tsvet had thus become the first person to separate natural pigments successfully.<\/p>\n<p><a href=\"https:\/\/www.rct-online.de\/en\/filtration\/liquid-chromatography-columns-for-hplc-and-mplc\/30900\/medium-pressure-liquid-chromatography-column-for-analytical-and-preparative-separation\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-16673 size-full\" title=\"Medium Pressure Liquid Chromatography Column for Analytical and Preparative Separation\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2023\/06\/medium-pressure-liquid-chromatography-column-for-analytical-and-preparative-separation.png\" alt=\"Medium Pressure Liquid Chromatography Column for Analytical and Preparative Separation\" width=\"300\" height=\"203\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2023\/06\/medium-pressure-liquid-chromatography-column-for-analytical-and-preparative-separation.png 300w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2023\/06\/medium-pressure-liquid-chromatography-column-for-analytical-and-preparative-separation-110x75.png 110w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a>\u00a0 <a href=\"https:\/\/www.rct-online.de\/de\/filtration\/fluessig-chromatographie-saeulen-fuer-hplc-und-mplc\/chromatographie-saeule\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-8828\" title=\"Chromatography Column\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/chromatographie-saeule1-2.jpg\" alt=\"Chromatography column\" width=\"300\" height=\"209\" \/><\/a><\/p>\n<p style=\"text-align: justify;\">The name \u201cTsvet\u201d means \u201ccolour\u201d in Russian. In 1906, he coined the term \u201cchromatography\u201d from the Greek \u2013 combining <em>chr\u014dma<\/em> (colour) and <em>graphein<\/em> (to write). The word is best translated loosely as \u201ccolour writing\u201d. It has served as his lasting memorial ever since.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"The_Slow_Rise_of_Column_Chromatography\"><\/span>The Slow Rise of Column Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">At first, though, Tsvet\u2019s \u201cchromatographic adsorption analysis\u201d, as he called it himself, remained largely unknown, because he published exclusively in Russian. The mechanisms behind the separation were also far from understood. Around the same time, the German-Jewish biochemist Richard Willst\u00e4tter (1872 \u2013 1942) was investigating the structure of chlorophyll at the <a href=\"https:\/\/www.mpic.de\/3537786\/Overview\">Kaiser Wilhelm Institute for Chemistry<\/a> in Berlin. He succeeded in isolating the pigment and in determining its molecular formula. For this work on plant pigments, he received the Nobel Prize in Chemistry in 1915.<\/p>\n<p style=\"text-align: justify;\">The column chromatography described by Tsvet only became more widely used in the 1930s. The German biochemists Edgar Lederer (1908 \u2013 1988) and Richard Kuhn (1900\u20131967) developed it into a practical laboratory method in Heidelberg.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"Chromatography_Becomes_a_Standard_Across_Many_Fields\"><\/span>Chromatography Becomes a Standard Across Many Fields<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p style=\"text-align: justify;\">The final breakthrough for chromatography came from understanding the separation mechanism. This insight grew out of the work of two British chemists in the 1940s: Archer John Porter Martin (1910 \u2013 2002) and Richard Laurence Millington Synge (1914 \u2013 1994). Drawing on experience from liquid extraction, they discovered how the separation actually works. Chromatographic separation depends on how differently substances distribute themselves between the stationary and the mobile phase.<\/p>\n<div class=\"box success  \"><div class=\"box-inner-block\"><i class=\"fa tie-shortcode-boxicon\"><\/i>\n\t\t\tNo specific binding to the stationary phase is required. Instead, the process relies on a repeated cycle of absorption and desorption between hydrophilic and hydrophobic phases. The resulting partition chromatography opened up new applications.\n\t\t\t<\/div><\/div>\n<p style=\"text-align: justify;\">It was soon used to decode the sequences of peptides. In doing so, it laid one of the cornerstones of modern biochemistry and molecular biology.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"Size-Exclusion_Chromatography\"><\/span>Size-Exclusion Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">Size-exclusion chromatography separates molecules by size, as its name suggests. Depending on the solvent used, it is also known as <a href=\"https:\/\/www.rct-online.de\/magazin\/en\/gel-permeation-chromatography-in-polymer-analysis\/\">gel-permeation chromatography<\/a> or gel-filtration chromatography.<\/p>\n<figure id=\"attachment_8809\" aria-describedby=\"caption-attachment-8809\" style=\"width: 450px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-8809 size-full\" title=\"Schematic Diagram of Paper Chromatography\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/Chromatography_tank.svg_.jpg\" alt=\"Schematic diagram of paper chromatography\" width=\"450\" height=\"493\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/Chromatography_tank.svg_.jpg 450w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/Chromatography_tank.svg_-274x300.jpg 274w\" sizes=\"(max-width: 450px) 100vw, 450px\" \/><figcaption id=\"caption-attachment-8809\" class=\"wp-caption-text\"><center>Schematic Diagram of Paper Chromatography (1: lid, 2: paper as stationary phase, 3: solvent front, 4: solvent)<\/center><\/figcaption><\/figure>\n<p style=\"text-align: justify;\">The analytes are separated by size as they pass through a porous gel. Small molecules enter the gel particles more easily. They therefore travel a longer path and elute later than larger ones. This makes the method particularly suitable for analysing <a href=\"https:\/\/www.rct-online.de\/en\/tubes-hoses\/tubings-and-pipes-made-of-rigid-plastics\">plastics<\/a>, where substances differ more by size than by their chemistry.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"High-Performance_Liquid_Chromatography\"><\/span>High-Performance Liquid Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">Biochemistry and natural-product chemistry deal with large molecules. For these, a further development of partition chromatography came into play: <a href=\"https:\/\/www.rct-online.de\/magazin\/en\/hplc-a-method-for-biochemical-research\/\">high-performance liquid chromatography (HPLC)<\/a>. Here, the separation process is accelerated. Two factors are decisive. The first is the high pressure of up to 400 bar. Secondly the use of very small stationary-phase particles, which offer a large surface area.<\/p>\n<p style=\"text-align: justify;\">The peripheral equipment required for HPLC must meet equally high standards of pressure resistance and chemical inertness. The <a href=\"https:\/\/www.rct-online.de\/en\/tubes-hoses\">tubing<\/a> therefore has to be especially pressure-resistant. Suitable options include <a href=\"https:\/\/www.rct-online.de\/en\/tubes-hoses\/tubings-and-pipes-made-of-rigid-plastics\/peek-tubing\">PEEK tubing<\/a> as well as <a href=\"https:\/\/www.rct-online.de\/en\/tubes-hoses\/tubing-pipes-and-capillarys-made-of-metal\/titanium-and-stainless-steel-capillaries\">titanium and stainless steel capillaries<\/a>. These are joined using <a href=\"https:\/\/www.rct-online.de\/en\/tube-hose-connectors\/connectors-made-of-metal\/capillary-connectors-for-hplc\">capillary connectors<\/a>, which allow connection to columns, detectors and other instruments.<\/p>\n<p><a href=\"https:\/\/www.rct-online.de\/en\/tubes-hoses\/tubings-and-pipes-made-of-rigid-plastics\/peek-tubing\/28754\/peek-high-pressure-capillary-tube-solid-colour\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-8817 size-full\" title=\"PEEK High-Pressure Capillary Tubing \u2013 Single Colour\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/peek-hochdruck-kapillarschlauch.jpg\" alt=\"PEEK high-pressure capillary tubing, single colour\" width=\"300\" height=\"300\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/peek-hochdruck-kapillarschlauch.jpg 300w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/peek-hochdruck-kapillarschlauch-150x150.jpg 150w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a> <a href=\"https:\/\/www.rct-online.de\/en\/tube-hose-connectors\/connectors-made-of-plastic\/capillary-connectors-for-hplc\/28886\/high-pressure-straight-capillary-connector-made-of-peek\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-8818 size-full\" title=\"Straight High-Pressure Capillary Connector, PEEK\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/kapillar-verbinder-fuer-die-hplc-hochdruck.jpg\" alt=\"Straight high-pressure capillary connector, PEEK\" width=\"300\" height=\"300\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/kapillar-verbinder-fuer-die-hplc-hochdruck.jpg 300w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/kapillar-verbinder-fuer-die-hplc-hochdruck-150x150.jpg 150w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a><\/p>\n<p style=\"text-align: justify;\">This high-pressure technique boosts the performance of column chromatography. Separation by hydrophobicity is not the only option, however. The method also helps to separate molecules by other properties, such as size, charge or affinity for a specific binding partner.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"Thin-Layer_Chromatography\"><\/span>Thin-Layer Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">Paper chromatography was later superseded by another powerful method: thin-layer chromatography. The technique dates back to 1938 and the work of two largely unknown Russians, N. A. Izmailov and M. S. Shraiber. In the early 1960s, the German chemist and pharmacist Egon Peter Gustav Stahl (1924 \u2013 1986) refined it further. Working with the companies Merck (Darmstadt) and Degussa (Heidelberg), he established it as a near-universal analytical method for non-volatile substances. He also developed the technical equipment it required.<\/p>\n<p style=\"text-align: justify;\">Thin-layer chromatography is technically simple and quick to carry out. Instead of paper, silica gels are the preferred stationary phase, along with aluminium oxides and modified celluloses. The stationary phase is applied in a thin layer to a flat support, usually a glass plate. An inert adhesive and binder, such as gypsum, holds it in place. The layer is then dried.<\/p>\n<p style=\"text-align: justify;\">As in paper chromatography, the samples are applied to the dry, solid phase. The plate is then placed upright in a closed tank, with the solvent level well below the application point. Capillary forces draw the eluent up into the solid phase. This separates the substances according to how differently they adsorb to and desorb from that phase.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"Gas_Chromatography\"><\/span>Gas Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">In the late 1940s, the German physical chemist Erika Cremer (1900 \u2013 1996) built on partition chromatography with her doctoral student Fritz Prior (1921 \u2013 1996). Instead of a liquid, they used gases such as helium or nitrogen as the mobile phase. For the stationary phase, they used a high-boiling, viscous liquid on a porous, inert support \u2013 for example, silicone oils on silica gel. Their fundamental work led to a new type of column separation method: gas chromatography. It works for gaseous substances and for those that can be vaporised without decomposing.<\/p>\n<p style=\"text-align: justify;\"><a href=\"https:\/\/www.rct-online.de\/en\/tubes-hoses\/tubing-pipes-and-capillarys-made-of-metal\/titanium-and-stainless-steel-capillaries\/28812\/stainless-steel-capillary-1.4301\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-8820 size-full\" title=\"Stainless Steel Capillary 1.4301\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/edelstahl-kapillare1.jpg\" alt=\"Stainless steel capillary 1.4301\" width=\"300\" height=\"300\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/edelstahl-kapillare1.jpg 300w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/edelstahl-kapillare1-150x150.jpg 150w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a> <a href=\"https:\/\/www.rct-online.de\/en\/tube-hose-connectors\/connectors-made-of-metal\/capillary-connectors-for-hplc\/29243\/male-screw-made-of-stainless-steel-1.4571\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-8821 size-full\" title=\"Hollow Screw, Stainless Steel 1.4571\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/hohlschraube-aus-edelstahl.jpg\" alt=\"Hollow screw, stainless steel 1.4571\" width=\"300\" height=\"300\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/hohlschraube-aus-edelstahl.jpg 300w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/hohlschraube-aus-edelstahl-150x150.jpg 150w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a><\/p>\n<p style=\"text-align: justify;\">Its development into a powerful laboratory technique is closely linked to two British chemists: Archer John Porter Martin (1910 \u2013 2002) and Anthony Trafford James (1922 \u2013 2006). The 1950s also brought compatible detectors. These included the thermal conductivity detector, the flame ionisation detector, the electron capture detector and, finally, mass-spectrometric detectors. Together, they were decisive in establishing gas chromatography as a standard analytical and micro-preparative method.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"Ion-Exchange_Chromatography\"><\/span>Ion-Exchange Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">Ion-exchange chromatography, also called ion chromatography, separates charged molecules, including nucleic acids and proteins. It does so according to their net charge, which depends on their isoelectric point. Their charge determines whether anion-exchange or cation-exchange chromatography is used. The principle is straightforward. The molecules bind ionically to an oppositely charged stationary phase. They are then eluted again by counter-ions from the eluent, at a suitable pH or ionic strength. An <a href=\"https:\/\/www.rct-online.de\/en\/filtration\/ion-exchange-membranes\">ion exchange<\/a> therefore takes place.<\/p>\n<p style=\"text-align: justify;\">In anion-exchange chromatography, negatively charged molecules \u2013 such as nucleic acids or proteins \u2013 bind to a positively charged stationary phase, for example a diethylaminoethyl (DEAE) resin. In cation-exchange chromatography, positively charged ions such as metal cations bind to a negatively charged stationary phase. One of the first applications came in the 1940s: the separation of rare-earth metals. These are hard to tell apart because of their chemical similarity. Ion-exchange chromatography is still used today to obtain pure rare-earth metals. These are indispensable raw materials for the entire field of microelectronics.<\/p>\n<h3><span class=\"ez-toc-section\" id=\"Affinity_Chromatography\"><\/span>Affinity Chromatography<span class=\"ez-toc-section-end\"><\/span><\/h3>\n<p style=\"text-align: justify;\">Affinity chromatography is widely used in the life sciences. It was developed in the 1960s by the Spanish-American pharmacologist Pedro Cuatrecasas (born 1936) and the Israeli biochemist Meir Wilchek (born 1935). The method relies on the interactions between biomolecules and their ligands \u2013 for example, between an enzyme and its substrate, or between an antibody and an antigen. The biomolecule binds to its immobilised ligand. It is then eluted by deliberately changing conditions such as pH, salt concentration or the addition of competing ligands.<\/p>\n<figure id=\"attachment_8811\" aria-describedby=\"caption-attachment-8811\" style=\"width: 450px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-8811 size-full\" title=\"High-Pressure Tubing at the Pump of an HPLC System\" src=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/hochdruckschlaeuche-an-der-pumpe.jpg\" alt=\"High-pressure tubing at the pump of an HPLC system\" width=\"450\" height=\"300\" srcset=\"https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/hochdruckschlaeuche-an-der-pumpe.jpg 450w, https:\/\/www.rct-online.de\/magazin\/wp-content\/uploads\/2024\/06\/hochdruckschlaeuche-an-der-pumpe-300x200.jpg 300w\" sizes=\"(max-width: 450px) 100vw, 450px\" \/><figcaption id=\"caption-attachment-8811\" class=\"wp-caption-text\"><center>High-Pressure Tubing at the Pump of an HPLC System<\/center><\/figcaption><\/figure>\n<p style=\"text-align: justify;\">The method is also often used to purify genetically tagged proteins. These proteins carry a tag, such as a histidine tag or glutathione-S-transferase, that enables interaction with a specific solid phase. Affinity chromatography makes it possible to purify biomolecules very specifically, even in complex mixtures. It is also used to purify vaccines. Most of us have therefore benefited directly from the power of this technique at some point.<\/p>\n<h2><span class=\"ez-toc-section\" id=\"Ferdinand_Runges_Legacy_Today\"><\/span>Ferdinand Runge\u2019s Legacy Today<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p style=\"text-align: justify;\">More than 150 years ago, Ferdinand Runge presented his chemical coats of arms and independently grown images to the public. He explained their formation through chemical interactions. Little could he have realised that he stood at the start of a dramatic development in the separation of substances.<\/p>\n<p style=\"text-align: justify;\">Later generations of scientists took up his basic ideas and developed them further. Today, chromatography exists in many technically mature forms. It separates substances both on the micro scale, to analyse mixtures, and on the macro scale, to obtain pure substances. Without these modern chromatographic separation and purification methods, we would have to do without much that we value or even depend on. There would be no mobile phones, no specific vaccines and no specific medicines \u2013 and probably a great deal else besides.<\/p>\n<pre><strong>Image sources: <\/strong>\r\nFeatured image | \u00a9 sinhyu \u2013 stock.adobe.com\r\npH measurement | \u00a9 Kim \u2013 stock.adobe.com\r\nSchematic diagram of paper chromatography | \u00a9 Theresa Knott, CC BY-SA 3.0 &lt;https:\/\/creativecommons.org\/licenses\/by-sa\/3.0&gt;, via Wikimedia Commons\r\nHigh-pressure tubing at the pump of an HPLC system | \u00a9 vladim_ka \u2013 stock.adobe.com<\/pre>\n","protected":false},"excerpt":{"rendered":"<p>Where the history of chromatography begins remains uncertain. Nature itself performs the process: a chemical mixture separates into its individual components as it passes through an environment made of different materials. Around 1900, the American geochemist David Talbot Day (1859 \u2013 1925) observed one such case. He found that the components of crude oil, as &hellip;<\/p>\n","protected":false},"author":10,"featured_media":13088,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1,3028],"tags":[4261,3103,4264,4259,3108,4260,4263,4262],"class_list":["post-16666","post","type-post","status-publish","format-standard","has-post-thumbnail","","category-all-articles","category-alle-beitraege-en","tag-affinity-chromatography","tag-chromatography","tag-ferdinand-runge-en","tag-gas-chromatography","tag-high-performance-liquid-chromatography","tag-ion-exchange-chromatography","tag-size-exclusion-chromatography","tag-thin-layer-chromatography"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v23.1 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>Chromatography: The History of Separation Techniques<\/title>\n<meta name=\"description\" content=\"What is chromatography, and which separation methods are used today? 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