Chromatography HPLC and GLC - Column chromatography, HPLC, Chromatogram, Applications and GC

Sep 20, 2026

Uploaded by Ophelia

VCE Chemistry Unit 3 Area of Study 1 learning notes on chromatography, covering the core ideas behind column chromatography, HPLC, and GLC/GC. 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.

LanguageEnglish

CategoryExam

FormatPDF

Pages23

Reading Time58 min

AccessFree to download

Column chromatography

HPLC

The Chromatogram

HPLC Applications

HPLC Sample exam question

Answer

Separation of mixture

Watch this lesson online: [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) CHEMISTRY Unit 3, Area of Study 1: Chemical Analysis Chromatography HPLC and GLC VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive and up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 1 Column chromatography (The basis for HPLC and GLC) • In the simplest form, column chromatography is carried out in the laboratory in a glass column packed with a solid such as alumina, Al 2 O 3 . VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive and up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 2 HPLC • Separation occurssin a short column • Stationary phase: very Iine solid particles tightly packed eg silica • Mobile phase: liquid under high pressure to force its way through the solid VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive and up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 3 The Chromatogram Rt mAU Rt- retention time-determines sample identity Rt Area or height is proportional to the quantity of analyte . Injection time VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive a4nd up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 4 HPLC Applications • HPLC is very sensitive! Chemical polystyrenes dyes phthalates Pharmaceuticals Environmental polyaromatic hydrocarbons Inorganic ions herbicides tetracyclines corticosteroids antidepressants barbiturates Bioscience proteins peptides nucleotides Consumer Products lipids antioxidants sugars Clinical amino acids vitamins homocysteine VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive a5nd up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 5 HPLC Sample exam question HPLC 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? (VCAA June 2009 Q 5) VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive and up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 6 Answer The answer is A • Molecules B, C and D are all polar. To see this observe the –OH and –NH groups on the hydrocarbon chain of the molecules. • 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. VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive and up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 7 GC • Sample and the solvent it is dissolved in must be volatile as they are heated in an oven • Mobile phase: Inert gas • Stationary phase: porous beads coated with high bpt liquid VCE Chemisrtry Unit 3 AOS 1 © Ms Lanna Derry & Edrolo 2014 We do our best to make these slides comprehensive and up-to-date, however there We'd appreciate it if you pointed these out to us! may be errors. 8 • Most sensitive of all chromatography techniques • Used for testing urine samples for drugs • Compounds must be able to be vaporised without decomposing (usually low molec

FAQs

What determines sample identity and what indicates quantity in HPLC chromatograms?

Retention time (Rt) determines sample identity. Peak area or height is proportional to the quantity of analyte.

How do stationary phase and mobile phase interact in HPLC?

The stationary phase is very finely packed solid particles. The mobile phase is a liquid under high pressure that forces separation through the solid.

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?

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.

What conditions are required for compounds in GC and what is used to detect eluted components?

Samples and their solvent must be volatile because they are heated in an oven. Eluted components are detected using a flame ionisation detector.

Sep 20, 2026

Uploaded by Ophelia