[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"$fxLN3MUwXCdr5RPjwZYIDpOj8CHyjOmngWTgoKXPtZbg":3,"$f1KHuf8PYEWHAaQHKaztGSELDcDckbnwUJWc0HmTDN7M":32,"$f3Ft0rKFJHppdzE-vuveecxx1BUcg9iOlMLtyzf_MJDg":80},[4,8,12,16,20,24,28],{"title":5,"slug":6,"path":7},"About Microbeonline.com","about-microbeonline-com","\u002Fabout-microbeonline-com\u002F",{"title":9,"slug":10,"path":11},"About Me","about-me","\u002Fabout-microbeonline-com\u002Fabout-me\u002F",{"title":13,"slug":14,"path":15},"Advertise with Us","advertise-us","\u002Fadvertise-us\u002F",{"title":17,"slug":18,"path":19},"Privacy Policy","privacy-policy","\u002Fprivacy-policy\u002F",{"title":21,"slug":22,"path":23},"Abbreviations","abbreviations","\u002Fabbreviations\u002F",{"title":25,"slug":26,"path":27},"Microbes","microbes","\u002Fmicrobes\u002F",{"title":29,"slug":30,"path":31},"Books","recommended-books","\u002Frecommended-books\u002F",{"type":33,"data":34},"blog",{"slug":35,"title":36,"description":36,"seoTitle":37,"seoDescription":37,"author":38,"createdDate":39,"lastUpdatedDate":40,"draft":41,"category":42,"image":37,"body":43,"faq":44,"tags":45,"related":47},"chromatography-an-overview","Chromatography: An Overview",null,"Ashma Shrestha","2022-05-15","2026-07-05",false,"lab-equipment","Did you know that green is a mixture of yellow and blue? This theory can be demonstrated by the separation technique, **chromatography** which means colored writing. It is one of the newest techniques for separating, identifying, and purifying the mixture of components.\n\nThe [IUPAC defines it](https:\u002F\u002Fgoldbook.iupac.org\u002Fterms\u002Fview\u002FC01075) as a “physical method of separation in which the components to be separated are distributed between two phases; one of which is the stationary phase while the other is the mobile phase that moves in a definite direction.\n\n## Brief History\n\n### Early Discovery\n\n- The first use of the chromatography method was purely by dye chemists. In 1861, Friedrich Goppelsroder described a method of obtaining a colored ring of drops of inorganic compounds in the center of filter paper. He gave it the name “capillary analysis.”\n- However, Russian botanist Mikhail Semyonovich Tsvet was the first documented inventor of the chromatography technique. He discovered this technique while researching chlorophyll in 1900. He mentioned the details of the process and named it chromatography.\n\n### Further Modification\n\n- In 1941, Archer J.P Martin and Richard L.M Synge for the separation of amino acids in wool. They were awarded the [Nobel Prize in Chemistry in 1952 for discovering partition-chromatography.](https:\u002F\u002Fwww.nobelprize.org\u002Fprizes\u002Fchemistry\u002F1952\u002Fsummary\u002F)The technique was similar in principle to Tsvet’s approach.\n- Due to different defects in the silica used in his technique, Martin and his team used filter paper as the stationary medium This modification gave the technique more reproducibility. It also replaced the column used by Tsvet to a large extent. The method was termed **“paper chromatography.”**\n- Likewise, two Soviet pharmacists, Nikolay A. Izmaylov and Maria S. Shrayber distributed a thin layer of support material on a glass plate. The German Chemist Egon Stahl further studied the spectrum of its application. And the system was termed **“thin layer chromatography.”**\n- Similarly, chemist Erika Cremer defined a new chromatographic technique where the stationary phase was solid instead of liquid. The technique is termed **gas chromatography.**\n\n## Principle of Chromatography\n\nThe basic principle of the chromatography technique is the same even though it has various types. This technique consists of the **two-phase; stationary and mobile phase.** The stationary phase does not move. The desired mixture of compounds (analytes) is present in the mobile phase. The analytes and the mobile phase move around the stationary phase.\n\nThe different components of analytes react at different times with the stationary phase while navigating through the stationary phase. It is because of their polarity with the stationary phase. This reaction time is termed retention time. Since the various components have different retention times, separation of the components occurs at different levels. The separated components pass through a detector. Afterward, they are plotted in a chromatogram.\n\n![Chromatography techniques - Chromatography techniques](https:\u002F\u002Fassets.microbeonline.com\u002Fblogs\u002Fchromatography-pic1.png)Figure: Chromatography techniques\n\n## Types of Chromatography\n\nThere are eight common types of chromatography which are; paper, thin-layer, gas, column, ion exchange, affinity, high-performance liquid, and immunochromatography.\n\n1. **Paper chromatography** is a technique that uses paper sheets that are absorbent as the stationary phase through which the liquid sample passes. This type commonly applies to check the purities of chemicals in various laboratories, determine the level of drugs in humans and animals, etc.\n2. **Thin-layer chromatography** is the process that uses aluminum foils or glass coated with a thin layer of absorbent material like cellulose, silica, or gel as a stationary phase through which liquid mobile moves. This movement and affinity of components in the mixture with the stationery phase helps separate the components. It is helpful for various qualitative and quantitative analyses of different compounds.\n3. [Gas chromatography](\u002Fgas-chromatography-gc-principle-parts-and-uses\u002F) is the separation process that uses gas (carrier gas) as a stationary phase where the liquid or gas sample gets injected\u002Finserted. It is useful in the separation of volatile substances.\n4. **Column chromatography:** It is the technique that uses a column with absorbent material as a stationary phase where the mobile phase along with sample travel. The movement depends on the difference in the affinity of components of the sample with the stationary phase leading to the separation of compounds at different times.\n5. **Ion exchange chromatography** is the technique that separates charged ions or polar compounds based on ionic interactions between the stationary phase and the mixture present in the mobile phase. It is applicable in various fields and works for any compounds that are capable of ionization.\n6. **Affinity chromatography**: It is the method based on the specific binding reaction between an immobilized stationary phase and mobile phase with the mixture.\n7. [High-performance liquid chromatography (HPLC)](\u002Fhplc-high-performance-liquid-chromatography\u002F) is the method in which pumps help liquid solvent to pass in high pressure through the column with solid absorbent material (stationary phase).\n8. Immunochromatography or Lateral flow Immunoassay: It is the combination of immunoassay and chromatography where immobilized antigen\u002Fantibody in the nitrocellulose membrane captures antibody\u002Fantigen present in the sample when allowed to pass through it.\n\n## Applications\u002F Uses of Chromatography\n\nThis method has wide applications in various fields of science. Some of the applications of chromatography are as follows:\n\n1. Chromatographic techniques apply in quality control testing in various industries.\n2. **The food industry** uses different chromatographic techniques to detect the presence of **chemical additives in food**. This technique also helps in assessing the **nutritional value of food and the shelf life**.\n3. **Pharmaceutical industries** use these techniques to detect the **chemicals and traces of foreign substances in the drugs**. This method applies to **developing new drugs**too.\n4. The **chemical industries** detect the **quality of air, water, pesticides, and impurities in petroleum products and oils** using this technique.\n5. **Forensic science** also uses this technique to **analyze various evidence.**\n6. This technique also applies in **molecular science** to separate**various proteins and nucleic acid components.**\n\n## Limitations\n\nAlthough chromatography has a wide spectrum of applications, it has few limitations. Some of its limitations are as follows:\n\n1. Chromatographic instruments and materials are expensive.\n2. Trained manpower is necessary for operating this technique.\n3. Maintenance and care of the equipment are necessary from time to time.\n4. Some chromatographic techniques may require a high load of electricity whereas others may require a high amount of solvents.\n\n**Reference**\n\n- MacNaught, A., & Wilkinson, A. (1997). *Compendium of chemical terminology*. Blackwell Science.\n- *Chromatography – New World Encyclopedia*. Newworldencyclopedia.org. (2022). Retrieved 11 May 2022, from \u003Chttps:\u002F\u002Fwww.newworldencyclopedia.org\u002Fentry\u002FChromatography>.\n- *Chromatography | Definition, Types, & Facts*. Encyclopedia Britannica. (2022). Retrieved 11 May 2022, from \u003Chttps:\u002F\u002Fwww.britannica.com\u002Fscience\u002Fchromatography>.\n- Coskun, O. (2016). Separation Techniques: CHROMATOGRAPHY. *Northern Clinics Of Istanbul*. \u003Chttps:\u002F\u002Fdoi.org\u002F10.14744\u002Fnci.2016.32757>.",[],[46],"chromatography",[48,55,61,67,73],{"slug":49,"title":50,"description":50,"seoTitle":37,"seoDescription":37,"author":51,"createdDate":52,"lastUpdatedDate":40,"draft":41,"category":42,"image":37,"faq":53,"tags":54},"hplc-high-performance-liquid-chromatography","High-Performance Liquid Chromatography (HPLC)","Sushmita Baniya","2022-07-01",[],[46],{"slug":56,"title":57,"description":57,"seoTitle":37,"seoDescription":37,"author":51,"createdDate":58,"lastUpdatedDate":40,"draft":41,"category":42,"image":37,"faq":59,"tags":60},"gas-chromatography-gc-principle-parts-and-uses","Gas Chromatography (GC): Principle, Parts and Uses","2022-06-15",[],[46],{"slug":62,"title":63,"description":63,"seoTitle":37,"seoDescription":37,"author":51,"createdDate":64,"lastUpdatedDate":40,"draft":41,"category":42,"image":37,"faq":65,"tags":66},"gel-filtration-chromatography","Gel Filtration Chromatography","2022-05-19",[],[46],{"slug":68,"title":69,"description":69,"seoTitle":37,"seoDescription":37,"author":51,"createdDate":70,"lastUpdatedDate":40,"draft":41,"category":42,"image":37,"faq":71,"tags":72},"thin-layer-chromatography","Thin Layer Chromatography (TLC): Principle, Procedure, and Applications","2022-05-17",[],[46],{"slug":74,"title":75,"description":75,"seoTitle":37,"seoDescription":37,"author":76,"createdDate":77,"lastUpdatedDate":40,"draft":41,"category":42,"image":37,"faq":78,"tags":79},"immunochromatography-principle-application","Immunochromatography\u002F Lateral Flow Immunoassay: Principle and Uses","Acharya Tankeshwar","2020-03-25",[],[46],[81,87,93,97,101,105,110,115,119,123],{"slug":82,"name":76,"description":83,"image":84,"body":85,"postCount":86},"acharya-tankeshwar","Editor-in-chief","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Ftankeshwar-acharya-author-microbeonline.jpg","***Tankeshwar Acharya, MSc (Medical Microbiology)***\n\n*Tankeshwar Acharya is an Assistant Professor in the Department of Microbiology at Patan Academy of Health Sciences (PAHS), Nepal, where he has been teaching and practicing clinical microbiology for over 14 years. He is the founder of Microbe Online, one of the leading free microbiology education resources on the web, covering bacteriology, mycology, parasitology, immunology, and clinical laboratory diagnostics written from direct experience in both the classroom and the diagnostic laboratory.*",432,{"slug":88,"name":38,"description":89,"image":90,"body":91,"postCount":92},"ashma-shrestha","SEO Copywriter and Science Communicator\nKathmandu, Nepal","https:\u002F\u002Fassets.microbeonline.com\u002Fauthors\u002Fashma-shrestha.png","Ashma Shrestha holds a Master of Science in Medical Microbiology from the Institute of Science and Technology (IOST), Tribhuvan University, Nepal, where she developed a strong foundation in virology, molecular biology, and diagnostic microbiology.\n\nShe now works as an SEO Copywriter at Resolution Digital, where she combines her scientific training with research-driven content strategy. She is certified in Google Analytics and Google Business Profile (GBP), and brings a data-informed approach to science communication writing content that is not only accurate but structured to reach and serve the students who need it most.\n\nAt microbeonline, Ashma contributes articles primarily in virology and molecular biology, areas she finds most compelling for their mechanistic depth and their growing clinical relevance. Her writing reflects the same standard the site is built on: factual rigor, clear explanation of the *why* behind microbiology concepts, and content that helps students move from memorization to genuine understanding.\n\nShe is passionate about making complex microbiological concepts accessible without sacrificing accuracy; a skill that sits at the intersection of her scientific training and her professional work in content and SEO.",81,{"slug":94,"name":51,"description":95,"image":37,"body":37,"postCount":96},"sushmita-baniya","Author \u002F Contributor",32,{"slug":98,"name":99,"description":95,"image":37,"body":37,"postCount":100},"samikshya-acharya","Samikshya Acharya",20,{"slug":102,"name":103,"description":95,"image":37,"body":37,"postCount":104},"alisha-tripathi","Alisha Tripathi",6,{"slug":106,"name":107,"description":108,"image":37,"body":37,"postCount":109},"aastha-shrestha","Aastha Shrestha"," Author \u002F Contributor",10,{"slug":111,"name":112,"description":113,"image":37,"body":37,"postCount":114},"guest-author","Guest Author","Guest Author \u002F Contributor",2,{"slug":116,"name":117,"description":95,"image":37,"body":37,"postCount":118},"srijana-khanal","Srijana Khanal",18,{"slug":120,"name":121,"description":113,"image":37,"body":37,"postCount":122},"dr-poonam-acharya","Dr. Poonam Acharya",1,{"slug":124,"name":125,"description":95,"image":37,"body":126,"postCount":127},"nisha-rijal","Nisha Rijal","**Nisha Rijal** is a microbiologist and quality assurance specialist. She served for nearly 12 years as a microbiologist at the National Public Health Laboratory (NPHL), Nepal's national reference laboratory, and continues to work as a consultant microbiologist in international public health organization. ",51]