{"id":468,"date":"2021-01-11T17:17:53","date_gmt":"2021-01-11T17:17:53","guid":{"rendered":"https:\/\/laboratoryinfo.com\/?p=468"},"modified":"2022-03-09T17:51:14","modified_gmt":"2022-03-09T17:51:14","slug":"hplc","status":"publish","type":"post","link":"https:\/\/laboratoryinfo.com\/hplc\/","title":{"rendered":"High Performance Liquid Chromatography (HPLC) : Principle, Types, Instrumentation and Applications"},"content":{"rendered":"\n<p>High-performance liquid chromatography, abbreviated as HPLC, is a chromatographic technique of great versatility and analytic power used in many aspects of drug manufacturing and research.<\/p>\n\n\n\n<p>It separates or identifies mixtures of substances into their components based on their molecular structure and composition. <\/p>\n\n\n\n<p>The other name for high-performance liquid chromatography is high-pressure liquid chromatography.&nbsp;<\/p>\n\n\n<style>.kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-table-of-content-wrap{padding-top:var(--global-kb-spacing-sm, 1.5rem);padding-right:var(--global-kb-spacing-sm, 1.5rem);padding-bottom:var(--global-kb-spacing-sm, 1.5rem);padding-left:var(--global-kb-spacing-sm, 1.5rem);background-color:#ecf2f7;}.kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-table-of-contents-title-wrap{padding-top:0px;padding-right:0px;padding-bottom:0px;padding-left:0px;}.kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-table-of-contents-title{font-weight:regular;font-style:normal;}.kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-table-of-content-wrap .kb-table-of-content-list{font-weight:regular;font-style:normal;margin-top:var(--global-kb-spacing-sm, 1.5rem);margin-right:0px;margin-bottom:0px;margin-left:0px;}.kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-toggle-icon-style-basiccircle .kb-table-of-contents-icon-trigger:after, .kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-toggle-icon-style-basiccircle .kb-table-of-contents-icon-trigger:before, .kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-toggle-icon-style-arrowcircle .kb-table-of-contents-icon-trigger:after, .kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-toggle-icon-style-arrowcircle .kb-table-of-contents-icon-trigger:before, .kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-toggle-icon-style-xclosecircle .kb-table-of-contents-icon-trigger:after, .kb-table-of-content-nav.kb-table-of-content-id6113_3b9ebd-8c .kb-toggle-icon-style-xclosecircle .kb-table-of-contents-icon-trigger:before{background-color:#ecf2f7;}<\/style>\n\n\n<h2 class=\"wp-block-heading\">Introduction<\/h2>\n\n\n\n<p>High-performance liquid chromatography (HPLC) is the most widely used separation technique. It can be very sensitive, specific, and precise. <\/p>\n\n\n\n<ul><li>It is a particular form of column chromatography used in biochemistry and analysis to separate, identify, and quantify the active compounds in a mixture.&nbsp;<\/li><li>In HPLC, a column holds packing material (stationary phase), a pump moves the mobile phase(s) through the column, and a detector shows the retention times of the molecules. <\/li><li>Retention time is variable and mainly depends on the interactions between the stationary phase, the molecules being analyzed, and the solvent(s) used.&nbsp;<\/li><li>A small volume of sample to be analyzed is introduced to the mobile phase stream and is retarded by specific chemical or physical interactions with the stationary phase. <\/li><li>The amount of retardation mainly depends on the nature of the analyte and the composition of both stationary and mobile phases. <\/li><li>The most common solvents used in high-performance liquid chromatography (HPLC) are methanol and acetonitrile.&nbsp;<\/li><\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Brief history<\/h2>\n\n\n\n<blockquote class=\"wp-block-quote\"><p>Michael Tswett (1872-1920) is credited as the father of chromatography due to his demonstration of liquid chromatography. In 1903, he separated the green-leaf pigments into bands of colors.&nbsp; After that, in 1937-38, thin-layer chromatography (TLC) was used. The next significant advancement was the use of <strong><a href=\"https:\/\/laboratoryinfo.com\/paper-chromatography\/\" target=\"_blank\" rel=\"noreferrer noopener\">paper chromatography<\/a><\/strong> in the mid-1940s.<\/p><\/blockquote>\n\n\n\n<p>Thin-layer chromatography (TLC) advanced slowly during the next few years, but Egon Stahl made significant development in 1956. Egon Stahl standardized the preparation of the sorbents used to make the plates.&nbsp; High-pressure liquid chromatography (HPLC) was later developed in the 1970s.&nbsp;<\/p>\n\n\n\n<p>The term high-performance liquid chromatography (HPLC) was introduced in the 1970s to distinguish the modern high-performance technique from classical low-pressure column chromatography, developed in the 1930s.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">HPLC Principle<\/h2>\n\n\n\n<p>High-performance liquid chromatography (HPLC) involves the injection of a small volume of liquid sample into a tube packed with tiny particles (3 to 5 microns (\u00b5m) in diameter called the stationary phase) where individual components of the sample are moved down the packed tube with a liquid (mobile phase) forced through the column by high pressure delivered through a pump.<\/p>\n\n\n\n<p>The column packing is used to separate the components from one another. It involves various chemical and\/or physical interactions between their molecules and the packing particles. <\/p>\n\n\n\n<p>The separated components are then detected at the exit of the column by a detector that measures their amount. Output from this detector is called a \u201c<strong>liquid chromatogram.<\/strong>\u201d<\/p>\n\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-full\"><img decoding=\"async\" width=\"968\" height=\"352\" src=\"http:\/\/laboratoryinfo.com\/wp-content\/uploads\/2021\/01\/Figure-1.1-Describes-the-basic-principle-of-HPLC.jpg\" alt=\"basic principle of High-performance liquid chromatography (HPLC)\" class=\"wp-image-7468\" srcset=\"https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2021\/01\/Figure-1.1-Describes-the-basic-principle-of-HPLC.jpg 968w, https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2021\/01\/Figure-1.1-Describes-the-basic-principle-of-HPLC-300x109.jpg 300w, https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2021\/01\/Figure-1.1-Describes-the-basic-principle-of-HPLC-768x279.jpg 768w\" sizes=\"(max-width: 968px) 100vw, 968px\" \/><figcaption>Figure: 1 Describes the basic principle of HPLC. <strong>Step 1<\/strong>. The sample is introduced (mobile phase). <strong>Step 2. <\/strong>The sample is separated into its components(stationary phase). <strong>Step 3<\/strong>. The sample is separated into its components (mobile phase).<\/figcaption><\/figure><\/div>\n\n\n\n<p><\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Advantages over low-pressure column liquid chromatography<\/h2>\n\n\n\n<p>There are many advantages of High-performance liquid chromatography (HPLC) over traditional low-pressure column liquid chromatography.&nbsp;<\/p>\n\n\n\n<ul><li>Greater sensitivity (various detectors can be employed)<\/li><li>Improved resolution<\/li><li>Speed&nbsp;<\/li><li>Easy sample recovery (less eluent volume to remove)<\/li><li>A wide variety of stationary phases<\/li><\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Branches of HPLC<\/h2>\n\n\n\n<p>Chromatography is divided into gas, liquid, and supercritical fluid techniques. <strong><a href=\"https:\/\/laboratoryinfo.com\/gas-chromatography-principle-applications-procedure-and-diagrams\/\" target=\"_blank\" rel=\"noreferrer noopener\">Gas chromatography<\/a><\/strong> is further divided into gas-liquid and gas-solid techniques. <\/p>\n\n\n\n<p>Liquid chromatography is divided into a relatively large collection of techniques like thin layer chromatography. Pressurized liquid chromatography can be divided into ion exchange, exclusion, partition, and liquid-solid chromatography.&nbsp;<\/p>\n\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-full\"><img decoding=\"async\" width=\"723\" height=\"476\" src=\"http:\/\/laboratoryinfo.com\/wp-content\/uploads\/2022\/03\/Describes-the-types-Branches-of-Chromatography-hplc.jpg\" alt=\"types Branches of Chromatography hplc High-performance liquid chromatography\" class=\"wp-image-7505\" srcset=\"https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2022\/03\/Describes-the-types-Branches-of-Chromatography-hplc.jpg 723w, https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2022\/03\/Describes-the-types-Branches-of-Chromatography-hplc-300x198.jpg 300w\" sizes=\"(max-width: 723px) 100vw, 723px\" \/><figcaption><strong><em>Figure 2.<\/em><\/strong><em> Describes the Branches of Chromatography.<\/em><\/figcaption><\/figure><\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Types of HPLC<\/h2>\n\n\n\n<p>The following variants of HPLC depend upon the phase system (stationary) in the process.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">1. Normal Phase HPLC<\/h3>\n\n\n\n<ul><li>They are also known as normal-phase or absorption chromatography. This method separates analytes based on polarity. <\/li><li>It has a polar stationary phase and a non-polar mobile phase. <\/li><li>Therefore, the stationary phase is usually silica, and typical mobile phases are hexane, methylene chloride, chloroform, diethyl ether, and mixtures. <\/li><li>The technique is used for water-sensitive compounds, geometric isomers, cis-trans isomers, class separations, and chiral compounds.&nbsp;<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">2. Reverse Phase HPLC<\/h3>\n\n\n\n<ul><li>The stationary phase is nonpolar (hydrophobic), while the mobile phase is an aqueous, moderate polar. <\/li><li>It works on the principle of hydrophobic interactions; hence the more nonpolar the material is, the longer it will be retained. <\/li><li>This technique is used for non-polar, polar, ionizable, and ionic molecules.&nbsp;<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">3. Size-exclusion HPLC<\/h3>\n\n\n\n<ul><li>It is also known as gel permeation chromatography or gel filtration chromatography. <\/li><li>The column is filled with a material having precisely controlled pore sizes, and the particles are separated according to their molecular size. <\/li><li>Larger molecules are rapidly washed through the column; smaller molecules penetrate the porous packing particles and elute later. <\/li><li>Size-exclusion chromatography is also helpful in determining the tertiary and quaternary structure of proteins and amino acids. <\/li><li>It is also used for the determination of the molecular weight of polysaccharides.<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">4. Ion-Exchange HPLC<\/h3>\n\n\n\n<ul><li>In this type of chromatography, retention is based on the attraction between solute ions and charged sites bound to the stationary phase. <\/li><li>Same charged ions are excluded. <\/li><li>This technique is used in purifying water, Ligand and Ion-exchange chromatography of proteins, high-pH anion-exchange chromatography of carbohydrates and oligosaccharides, etc.&nbsp;<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">5. Bio-affinity HPLC<\/h3>\n\n\n\n<ul><li>In this type of chromatography, separation is based on the reversible interaction of proteins with ligands.&nbsp;<\/li><\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Instrumentation of HPLC<\/h2>\n\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-full\"><img decoding=\"async\" width=\"800\" height=\"295\" src=\"http:\/\/laboratoryinfo.com\/wp-content\/uploads\/2015\/07\/High-performance-liquid-chromatography-hplc.jpg\" alt=\"High performance liquid chromatography hplc Instrumentation parts mechanism\" class=\"wp-image-469\" srcset=\"https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2015\/07\/High-performance-liquid-chromatography-hplc.jpg 800w, https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2015\/07\/High-performance-liquid-chromatography-hplc-300x111.jpg 300w, https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2015\/07\/High-performance-liquid-chromatography-hplc-768x283.jpg 768w, https:\/\/laboratoryinfo.com\/wp-content\/uploads\/2015\/07\/High-performance-liquid-chromatography-hplc-20x7.jpg 20w\" sizes=\"(max-width: 800px) 100vw, 800px\" \/><figcaption><em>The schematic diagram the above figure shows that the basic HPLC system consists of a pump, injector, column, detector, and integrator or acquisition and display system. <\/em><br><em><strong>The heart of the system is the column where separation occurs.<\/strong><\/em><\/figcaption><\/figure><\/div>\n\n\n\n<p class=\"has-text-align-center\"><br><\/p>\n\n\n\n<h3 class=\"wp-block-heading\">1. Solvent Reservoir<\/h3>\n\n\n\n<ul><li>Mobile phase contents are contained in a glass reservoir. <\/li><li>The mobile phase, or solvent, in HPLC, is usually a mixture of polar and non-polar liquid components whose respective concentrations are varied depending on the composition of the sample.<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">2. Pump<\/h3>\n\n\n\n<ul><li>A pump aspirates the mobile phase from the solvent reservoir and forces it through the system\u2019s column and detecter. <\/li><li>Depending on several factors, including column dimensions, the particle size of the stationary phase, the flow rate and composition of the mobile phase, operating pressures of up to 42000 kPa (about 6000 psi) can be generated.<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">3. Sample Injector<\/h3>\n\n\n\n<ul><li>The injector can be a single injection or an automated injection system. <\/li><li>An injector for an HPLC system should provide an injection of the liquid sample within the range of 0.1-100 mL of volume with high reproducibility and under high pressure (up to 4000 psi).<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">4. Columns<\/h3>\n\n\n\n<ul><li>Columns are usually made of polished stainless steel, are between 50 and 300 mm long, and have an internal diameter between 2 and 5 mm. <\/li><li>They are commonly filled with a stationary phase with a particle size of 3\u201310 \u00b5m. <\/li><li>Columns with internal diameters of less than 2 mm are often called microbore columns. <\/li><li>Ideally, the temperature of the mobile phase and the column should be kept constant during an analysis.<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">5. Detector<\/h3>\n\n\n\n<ul><li>The HPLC detector, located at the end of the column, detects the analytes as they elute from the chromatographic column. <\/li><li>Commonly used detectors are UV-spectroscopy, fluorescence, mass-spectrometric and electrochemical detectors.<\/li><\/ul>\n\n\n\n<h3 class=\"wp-block-heading\">6. Data Collection Devices<\/h3>\n\n\n\n<ul><li>Signals from the detector may be collected on chart recorders or electronic integrators that vary in complexity and their ability to process, store and reprocess chromatographic data. <\/li><li>The computer integrates the detector&#8217;s response to each component and places it into a chromatograph that is easy to read and interpret.<\/li><\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Applications of HPLC<\/h2>\n\n\n\n<p>The information that HPLC can obtain includes resolution, identification, and quantification of a compound. It also aids in chemical separation and purification. The other applications of HPLC include<\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Pharmaceutical Applications<\/h3>\n\n\n\n<ol><li>To control drug stability.<\/li><li>Tablet dissolution study of pharmaceutical dosages form.<\/li><li>&nbsp;Pharmaceutical quality control.<\/li><\/ol>\n\n\n\n<h3 class=\"wp-block-heading\">Environmental Applications<\/h3>\n\n\n\n<ol><li>Detection of phenolic compounds in drinking water.<\/li><li>Bio-monitoring of pollutants.<\/li><\/ol>\n\n\n\n<h3 class=\"wp-block-heading\">Applications in Forensics<\/h3>\n\n\n\n<ol><li>Quantification of drugs in biological samples.<\/li><li>Identification of steroids in blood, urine, etc.<\/li><li>Forensic analysis of textile dyes.<\/li><li>Determination of cocaine and other drugs of abuse in blood, urine, etc.<\/li><\/ol>\n\n\n\n<h3 class=\"wp-block-heading\">Food and Flavour<\/h3>\n\n\n\n<ol><li>Measurement of Quality of soft drinks and water.<\/li><li>Sugar analysis in fruit juices.<\/li><li>Analysis of polycyclic compounds in vegetables.<\/li><li>Preservative analysis.<\/li><\/ol>\n\n\n\n<h3 class=\"wp-block-heading\">Applications in Clinical Tests<\/h3>\n\n\n\n<ol><li>Urine analysis, antibiotics analysis in blood.<\/li><li>Analysis of bilirubin, biliverdin in hepatic disorders.<\/li><li>Detection of endogenous Neuropeptides in the extracellular fluid of the brain etc.<\/li><\/ol>\n\n\n\n<h2 class=\"wp-block-heading\">Limitations<\/h2>\n\n\n\n<p>The limitation of using high-performance liquid chromatography (HPLC) is the following.<\/p>\n\n\n\n<ul><li>HPLC is much more costly requires a large number of expensive organics.<\/li><li>HPLC may have low sensitivity for certain compounds, and some cannot even be detected as they are irreversibly adsorbed.<\/li><li>Complexity<\/li><li>Volatile substances are much better to be separated by gas chromatography.<\/li><\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Frequently asked Questions<\/h2>\n\n\n<div id=\"rank-math-faq\" class=\"rank-math-block\">\n<div class=\"rank-math-list \">\n<div id=\"faq-question-1621931861282\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 1. What is liquid chromatography?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>Liquid chromatography is one of the three main branches of chromatography. It involves a small volume of liquid sample placement into a tube packed with porous particles. <br \/>The individual components of the sample are transported along the column by a liquid moved with gravity. <br \/>The sample components are separated and then collected at the exit of this column.\u00a0<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932091903\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 2. What is the principle of HPLC?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>The principle of HPLC is based on analyte distribution between the mobile and stationary phases. It is crucial to remember that the sample\u2019s different constituents elute at various times before the sample ingredients\u2019 separation is achieved. The intermolecular interactions between sample and packaging materials molecules determine their time on-column.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932225527\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 3. What are the types of HPLC?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>There are four primary types of HPLC<br \/>1. <strong>Normal phase HPLC <\/strong>(effective method for separating phospholipid classes)<br \/>2. <strong>Reverse phase HPLC<\/strong> (the most common method used to separate compounds that have hydrophobic moieties)<br \/>3. <strong>Size-exclusion HPLC\/molecular sieve chromatography <\/strong>(Used in large molecules\/macromolecular complexes such as industrial polymers and proteins)<br \/>4. <strong>Ion-exchange HPLC <\/strong>(separates ions and polar molecules according to their ion exchanger.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932232606\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 4. What are the four types of chromatography?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>The four types of chromatography are<br \/>1. <strong>Liquid chromatography <\/strong>(test for pollution in water samples like lakes and rivers)<br \/>2. <strong>Gas chromatography<\/strong> (detect bombs and valuable in forensic investigations)<br \/>3. <strong>Thin-layer chromatography<\/strong> (used to check the purity of organic compounds such as the presence of insecticide or pesticide in foods)<br \/>4. <strong>Paper chromatography<\/strong> (uses a strip of paper in the stationary phase).<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932238388\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q5. Why is high pressure needed in HPLC?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>HPLC uses a moderate to high pressure to achieve the desired flow rate of the solvent through the chromatographic column as small particles have more excellent resistance to flow.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932261613\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 6. What is the difference between isocratic and gradient?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>Your application can be run in different ways \u2013 isocratic and gradient. <br \/><strong>Isocratic <\/strong>is when the mobile phase mixture is consistent over the total testing time. <br \/>With a <strong>gradient<\/strong>, the compounding of the eluent mixture is changed during measurement, which significantly affects analyte retention. It can accelerate or decelerate the separation process.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932268342\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 7. What solvent is used in HPLC?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>Different solvents are used in HPLC, such as aqueous solvent (water) and organic solvent (methanol, acetonitrile, and propanol). To improve the chromatographic peak shape, acids such as acetic acid, formic acid, and trifluoroacetic acid can be used.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932276495\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 8. What is the difference between UV and PDA detectors?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>The PDA and UV are both absorbance detectors, which provide sensitivity for light-absorbing compounds. The UV detector is most commonly used for HPLC analysis. <br \/>The UV absorbance differs on the wavelength used, so it is essential to choose the right wavelength based on the type of analyte. On the other hand, the PDA detector adds a third dimension wavelength, which is a more convenient way of finding out the wavelength without repeating the analysis.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932285192\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 9. What are the advantages of HPLC?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>The advantages of HPLC are as follows:<br \/>1.\u00a0 It can test both raw materials and finished products.<br \/>2.\u00a0 It can reverse engineer formulations.<br \/>3.\u00a0 It helps solve product failure problems.<br \/>4.\u00a0 It can detect contaminants and other impurities.<br \/>5.\u00a0 It can perform competitor product analysis.<br \/>6.\u00a0 It can determine product stability and shelf life.<br \/>7.\u00a0 The testing can be done even with just a small sample size.<br \/>8.\u00a0 It enables you to modify the testing depending on the needed quantification level.<br \/>9.\u00a0 The results it produced are reliable.<br \/>10. It helps develop better products.<br \/>11. It\u00a0lets you gain a better understanding of the competitor\u2019s products.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1621932299079\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 10. What is Rf value?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>In chromatography, the RF value pertains to the distance a particular component traveled divided by the distance traveled by the solvent front. In other words, it is the characteristic of the component which is helpful in the identification of the components.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1646414428512\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 11. What are the two main types of chromatography?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>There are different types of chromatography, but the two primary types are liquid chromatography and gas chromatography.<\/p>\n\n<\/div>\n<\/div>\n<div id=\"faq-question-1646414460301\" class=\"rank-math-list-item\">\n<h3 class=\"rank-math-question \">Q 12. What is retention time?<\/h3>\n<div class=\"rank-math-answer \">\n\n<p>It is when a specific analyte comes out of the end of the column.<\/p>\n\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n\n\n<p><strong>References:<\/strong><\/p>\n\n\n\n<ol><li><em>Malviya, Rishabha &amp; Bansal, Vvipin &amp; Pal, Om &amp; Sharma, Pramod. (2010). High-performance liquid chromatography: A short review. Journal of Global Pharma Technology. 2. 22-26.&nbsp;<\/em><\/li><li><em>https:\/\/microbenotes.com\/high-performance-liquid-chromatography-hplc\/<\/em><\/li><li><em>https:\/\/www.chemguide.co.uk\/analysis\/chromatography\/hplc.html<\/em><\/li><li><em><a href=\"https:\/\/www2.chemistry.msu.edu\/courses\/cem434\/Swain_2015_Lecture%20Notes\/HPLC%20Lecture1a.pdf\" rel=\"nofollow noopener\" target=\"_blank\">https:\/\/www2.chemistry.msu.edu\/courses\/cem434\/Swain_2015_Lecture%20Notes\/HPLC%20Lecture1a.pdf<\/a><\/em><\/li><li><em>https:\/\/www.nature.com\/subjects\/liquid-chromatography<\/em><\/li><li><em>https:\/\/www.knauer.net\/en\/Systems-Solutions\/Analytical-HPLC-UHPLC\/HPLC-Basicsprinciples-and-parameters<\/em><\/li><li><em>https:\/\/www.azom.com\/article.aspx?ArticleID=8468<\/em><\/li><li><em><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC5702474\/\" rel=\"nofollow noopener\" target=\"_blank\">https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC5702474\/<\/a><\/em><\/li><\/ol>\n\n\n\n<p><\/p>\n","protected":false},"excerpt":{"rendered":"<p>High-performance liquid chromatography, abbreviated as HPLC, is a chromatographic technique of great versatility and analytic power used in many aspects of drug manufacturing and research. It separates or identifies mixtures of substances into their components based on their molecular structure and composition. The other name for high-performance liquid chromatography is high-pressure liquid chromatography.&nbsp; Introduction High-performance&#8230;<\/p>\n","protected":false},"author":4,"featured_media":469,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"rank_math_lock_modified_date":false,"_kad_post_transparent":"","_kad_post_title":"","_kad_post_layout":"","_kad_post_sidebar_id":"","_kad_post_content_style":"","_kad_post_vertical_padding":"","_kad_post_feature":"","_kad_post_feature_position":"","_kad_post_header":false,"_kad_post_footer":false,"footnotes":""},"categories":[2],"tags":[31,40],"acf":[],"_links":{"self":[{"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/posts\/468"}],"collection":[{"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/users\/4"}],"replies":[{"embeddable":true,"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/comments?post=468"}],"version-history":[{"count":0,"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/posts\/468\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/media\/469"}],"wp:attachment":[{"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/media?parent=468"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/categories?post=468"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/laboratoryinfo.com\/wp-json\/wp\/v2\/tags?post=468"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}