Chromatography Systems Basics Course

Lesson 1 • Core Separation Principles

Explains partitioning, adsorption, and differential migration as the basis of all separations. Links these mechanisms to observable chromatographic behaviour.

Lesson 2 • Band Broadening and Efficiency

Introduces the van Deemter equation and plate theory to explain peak width. Connects efficiency metrics to practical column and flow-rate choices.

Lesson 3 • Stationary and Mobile Phases

Defines the roles of stationary and mobile phases in controlling selectivity. Introduces polarity, viscosity, and compatibility as phase-selection criteria.

Lesson 4 • Retention and Selectivity Concepts

Quantifies how analytes interact with phases using retention factors and selectivity ratios. Builds the vocabulary needed for method development discussions.

Lesson 1 • Data Acquisition and System Control

Explains analog-to-digital conversion, chromatography data systems, and instrument control software. Links hardware signals to digital chromatograms.

Lesson 2 • Detectors and Signal Transduction

Surveys UV-Vis, fluorescence, refractive index, and mass spectrometric detectors. Explains how each transducer converts analyte presence into an electrical signal.

Lesson 3 • Sample Introduction Devices

Describes manual injectors, autosamplers, and headspace samplers used to introduce analytes. Connects injection volume and technique to reproducibility.

Lesson 4 • Columns and Column Hardware

Examines column geometry, packing materials, and fittings that define separation performance. Introduces guard columns and column switching configurations.

Lesson 1 • Resolution and Peak Quality in HPLC

Applies the resolution equation to diagnose and improve peak separation. Connects plate count, selectivity, and retention to practical resolution strategies.

Lesson 2 • HPLC System Overview and Modes

Introduces reversed-phase, normal-phase, and ion-exchange HPLC as distinct separation strategies. Connects analyte polarity and ionisation state to mode selection.

Lesson 3 • Mobile Phase Composition and Gradients

Covers solvent strength, buffer selection, and gradient programming for HPLC. Explains how gradient shape affects resolution and run time.

Lesson 4 • HPLC Method Parameters and Setup

Defines flow rate, column temperature, injection volume, and detection wavelength as key run parameters. Demonstrates systematic parameter adjustment for peak optimisation.

Lesson 1 • GC Detectors and Their Selectivity

Surveys FID, TCD, ECD, NPD, and FPD detectors with their selectivity and sensitivity profiles. Matches detector choice to analyte class and regulatory requirements.

Lesson 2 • Temperature Programming in GC

Explains isothermal and temperature-programmed runs and their effect on elution order. Demonstrates ramp rate optimisation for complex mixtures.

Lesson 3 • GC System Architecture

Maps the flow path from carrier gas supply through injector, column oven, and detector. Establishes how each component contributes to separation quality.

Lesson 4 • Injection Techniques in GC

Compares split, splitless, on-column, and programmed-temperature vaporisation injection modes. Links injection choice to analyte volatility, concentration, and matrix complexity.

Lesson 1 • Limits of Detection and Quantitation

Defines LOD and LOQ using signal-to-noise and regression approaches. Applies these limits to assess method suitability for target analyte concentrations.

Lesson 2 • Peak Identification Strategies

Uses retention time, relative retention, and spectral matching to confirm analyte identity. Introduces co-injection and spiking as confirmation tools.

Lesson 3 • Uncertainty and Result Reporting

Introduces measurement uncertainty propagation from calibration, integration, and sample preparation. Guides compliant result reporting with appropriate significant figures.

Lesson 4 • Calibration Methods and Models

Compares external standard, internal standard, and standard addition calibration approaches. Explains when each model is appropriate based on matrix and instrument variability.

Lesson 1 • Parameter Optimisation Strategies

Applies one-variable-at-a-time and multivariate optimisation to refine flow rate, temperature, and gradient. Balances resolution, run time, and sensitivity trade-offs.

Lesson 2 • Defining Method Goals and Analyte Properties

Establishes target analytes, concentration ranges, matrix types, and regulatory requirements before column selection. Prevents costly rework by aligning goals with technique capabilities.

Lesson 3 • Method Documentation and Transfer

Structures method documents with sufficient detail for successful transfer to another laboratory. Covers system suitability criteria and transfer acceptance criteria.

Lesson 4 • Robustness and Ruggedness Testing

Evaluates method performance under deliberate small variations in critical parameters. Identifies control limits that protect method performance during routine use.

Lesson 1 • Validation Framework and Terminology

Defines validation parameters—specificity, linearity, accuracy, precision, range, and robustness—within a regulatory context. Establishes the validation plan as the governing document.

Lesson 2 • Validation Report and Lifecycle Management

Compiles validation data into a structured report with conclusions and regulatory submission readiness. Introduces post-approval change control and revalidation triggers.

Lesson 3 • Precision: Repeatability and Reproducibility

Measures intra-day repeatability and inter-day intermediate precision using replicate injections. Applies %RSD criteria to confirm consistent method performance.

Lesson 4 • Linearity, Range, and Accuracy

Establishes the working range with regression analysis and evaluates accuracy through recovery experiments. Links these parameters to the method's intended concentration span.

Lesson 1 • Baseline and Noise Issues

Identifies sources of baseline drift, spikes, and high noise in detector and electronic systems. Guides corrective actions from mobile phase degassing to detector cleaning.

Lesson 2 • Systematic Troubleshooting Approach

Introduces a structured fault-isolation workflow from symptom observation to root-cause confirmation. Prevents misdiagnosis by separating instrument, column, and method variables.

Lesson 3 • Pressure and Flow Anomalies

Diagnoses high pressure, low pressure, and flow instability from pump, column, and connection faults. Provides step-by-step isolation procedures for each pressure scenario.

Lesson 4 • Common Peak Shape Problems

Diagnoses tailing, fronting, splitting, and ghost peaks using chromatographic evidence. Links each defect to specific hardware, column, or method causes.