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Explains how stationary and mobile phases operate, how retention time (Rt) and peak area relate to analyte identity and quantity, and how separation behavior links to polarity. Includes HPLC application categories such as pharmaceuticals, environmental compounds, bioscience, consumer products, and clinical substances, plus an exam-style question and answer rationale. Also outlines GC requirements and the detection principle during elution.",{"@graph":69,"@context":126},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":20,"@type":76,"position":81},"https://docshare.wps.com/document/exam/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/chromatography-hplc-and-glc-column-chromatography-hplc-chromatogram-applications-and-gc/315884/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/chromatography-hplc-and-glc-column-chromatography-hplc-chromatogram-applications-and-gc/315884.png","ImageObject",300,407,{"name":92,"@type":93},"Ophelia","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-11-03","2026-09-20",true,{"@type":102,"interactionType":103,"userInteractionCount":39},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118,122],{"name":109,"@type":110,"acceptedAnswer":111},"What determines sample identity and what indicates quantity in HPLC chromatograms?","Question",{"text":112,"@type":113},"Retention time (Rt) determines sample identity. Peak area or height is proportional to the quantity of analyte.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How do stationary phase and mobile phase interact in HPLC?",{"text":117,"@type":113},"The stationary phase is very finely packed solid particles. The mobile phase is a liquid under high pressure that forces separation through the solid.",{"name":119,"@type":110,"acceptedAnswer":120},"Why does a non-polar molecule show the greatest retention time on an HPLC column described as non-polar stationary with a polar mobile phase?",{"text":121,"@type":113},"A non-polar molecule has the greatest attraction to the non-polar stationary phase, so it moves more slowly through the column and shows the longest retention time.",{"name":123,"@type":110,"acceptedAnswer":124},"What conditions are required for compounds in GC and what is used to detect eluted components?",{"text":125,"@type":113},"Samples and their solvent must be volatile because they are heated in an oven. Eluted components are detected using a flame ionisation detector.","https://schema.org",{"og:url":83,"og:type":128,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":130,"canonical":83},"index,follow",{"doc_id":132,"site_id":62},315884,1790024738,{"code":4,"msg":5,"data":135},{"doc_id":132,"user_id":136,"nickname":92,"user_avatar":137,"doc_module":4,"category_id":19,"category_name":20,"doc_title":65,"doc_description":67,"doc_content":138,"file_id":139,"file_url":140,"file_type":141,"file_size":142,"view_count":39,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":143,"language":144,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":145,"faqs":146,"seo_title":147,"seo_description":67,"update_tm":148,"read_time":149,"preview_page_count":4},7971461741311,"https://ap-avatar.wpscdn.com/avatar/74000253aff267980c6?x-image-process=image/resize,m_fixed,w_180,h_180&k=1779345379180704826","Watch this lesson online:  \n[https://edrolo.com.au/vce/subjects/chemistry/vce-chemistry/aos-1-chemical-analysis/chromatography-hplc-glc/column-chromatography/\\#watch](https://edrolo.com.au/vce/subjects/chemistry/vce-chemistry/aos-1-chemical-analysis/chromatography-hplc-glc/column-chromatography/#watch)  \nCHEMISTRY  \nUnit 3, Area of Study 1: Chemical Analysis  \nChromatography HPLC and GLC  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive and up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n1  \nColumn chromatography  \n(The basis for HPLC and GLC)  \n• In the simplest form, column  \nchromatography is carried out in the laboratory in a glass column packed with a solid such as alumina, Al 2 O 3 .  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive and up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n2  \nHPLC  \n• Separation occurssin a short column  \n• Stationary phase:  \nvery Iine solid particles tightly packed eg silica  \n• Mobile phase: liquid under high pressure to force its way through the solid  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive and up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n3  \nThe Chromatogram  \nRt  \nmAU  \nRt- retention time-determines sample identity  \nRt  \nArea or height is proportional to the quantity of analyte .  \nInjection  \ntime  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive a4nd up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n4  \nHPLC Applications  \n• HPLC is very sensitive!  \nChemical  \npolystyrenes dyes phthalates  \nPharmaceuticals  \nEnvironmental  \npolyaromatic hydrocarbons Inorganic ions  \nherbicides  \ntetracyclines corticosteroids antidepressants barbiturates  \nBioscience  \nproteins peptides nucleotides  \nConsumer Products  \nlipids antioxidants sugars  \nClinical  \namino acids vitamins homocysteine  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive a5nd up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n5  \nHPLC Sample exam question  \nHPLC uses a n~~on~~-­‐~~polar stationary phase togeth~~er with a polar ~~mobile phase. ~~Which of the following molecules will have the greatest retention time on such a column?  \n(VCAA June 2009 Q 5)  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive and up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n6  \nAnswer  \nThe answer is A  \n• Molecules B, C and D are all polar. To see this observe the –OH and –NH groups on the hydrocarbon chain of the molecules.  \n• A non-­‐polar molecule will have the greatest attraction to the stationary phase, so will take the longest time to move along the column = greatest retention time.  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive and up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n7  \nGC  \n• Sample and the solvent it is dissolved in must be volatile as they are heated in an oven  \n• Mobile phase: Inert gas  \n• Stationary phase: porous beads coated with high bpt liquid  \nVCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014  \nWe do our best to make these slides comprehensive and up-to-date, however there  \nWe'd appreciate it if you pointed these out to us!  \nmay be errors.  \n8  \n• Most sensitive of all chromatography techniques  \n• Used for testing urine samples for drugs  \n• Compounds must be able to be vaporised without decomposing (usually low molec","cbCaigOsoFbJlD7D","https://ap.wps.com/l/cbCaigOsoFbJlD7D","pdf",493260,23,"English","# Column chromatography\n## HPLC\n## The Chromatogram\n## HPLC Applications\n## HPLC Sample exam question\n## Answer\n## GC\n## Separation of mixture","[{\"question\":\"What determines sample identity and what indicates quantity in HPLC chromatograms?\",\"answer\":\"Retention time (Rt) determines sample identity. Peak area or height is proportional to the quantity of analyte.\"},{\"question\":\"How do stationary phase and mobile phase interact in HPLC?\",\"answer\":\"The stationary phase is very finely packed solid particles. The mobile phase is a liquid under high pressure that forces separation through the solid.\"},{\"question\":\"Why does a non-polar molecule show the greatest retention time on an HPLC column described as non-polar stationary with a polar mobile phase?\",\"answer\":\"A non-polar molecule has the greatest attraction to the non-polar stationary phase, so it moves more slowly through the column and shows the longest retention time.\"},{\"question\":\"What conditions are required for compounds in GC and what is used to detect eluted components?\",\"answer\":\"Samples and their solvent must be volatile because they are heated in an oven. Eluted components are detected using a flame ionisation detector.\"}]","Chromatography HPLC and GLC - Column chromatography, HPLC, Chromatogram, Applications and GC | PDF",1789907547,58]