Liquid and Gas Chromatography Course

Lesson 1 • Resolution, Selectivity, and Efficiency

Defines resolution mathematically and decomposes it into efficiency, selectivity, and retention factor. Provides tools to diagnose and improve separation quality.

Lesson 2 • Band Broadening and Plate Theory

Introduces theoretical plates and the van Deemter equation to quantify column efficiency. Connects peak width to practical column and flow optimization.

Lesson 3 • Thermodynamic Basis of Retention

Explains partition, adsorption, and distribution coefficients governing analyte retention. Links Gibbs free energy changes to observed chromatographic behavior.

Lesson 4 • Stationary and Mobile Phase Interactions

Surveys polarity, hydrogen bonding, and dispersion forces that drive analyte partitioning. Establishes solvent strength and selectivity concepts used throughout the course.

Lesson 1 • Gas Chromatograph Architecture

Describes carrier gas supply, flow controllers, injectors, oven, column, and detector as an integrated system. Emphasizes how each component affects separation quality.

Lesson 2 • Data Acquisition and System Control

Covers analog-to-digital conversion, chromatography data systems, and instrument control software. Establishes data integrity practices required for regulated environments.

Lesson 3 • Liquid Chromatograph Architecture

Maps solvent reservoir, degasser, pump, injector, column compartment, and detector in an HPLC system. Highlights pressure ratings and flow path integrity.

Lesson 4 • HPLC Pump Technology

Explains reciprocating piston, syringe, and quaternary pump designs and their flow accuracy. Covers pulse dampening and gradient formation methods.

Lesson 1 • Column Care, Conditioning, and Troubleshooting

Details conditioning protocols, storage, and common failure modes such as contamination and breakage. Enables students to extend column lifetime and diagnose performance loss.

Lesson 2 • Packed Columns and Their Applications

Covers support materials, liquid loadings, and applications where packed columns outperform capillary columns. Addresses gas analysis and preparative-scale GC.

Lesson 3 • Capillary Column Design and Geometry

Explains fused-silica capillary construction, deactivation, and dimensional parameters. Links inner diameter, film thickness, and length to efficiency and capacity.

Lesson 4 • Column Selection Strategy

Provides a systematic workflow for matching column polarity, dimensions, and temperature range to target analytes. Integrates Kovats retention index data into selection.

Lesson 1 • Thermal Conductivity Detector Operation

Describes Wheatstone bridge design and carrier gas thermal conductivity principles. Positions TCD for universal detection of inorganic and permanent gases.

Lesson 2 • Detector Performance Metrics and Selection

Defines sensitivity, selectivity, linearity, minimum detectable quantity, and dynamic range. Guides detector choice based on analyte matrix and regulatory detection limits.

Lesson 3 • Selective Detectors: ECD, NPD, and FPD

Covers electron capture, nitrogen-phosphorus, and flame photometric detectors for targeted analyte classes. Addresses radioactive source handling and detector-specific calibration.

Lesson 4 • Mass Spectrometric Detection in GC

Introduces quadrupole, ion trap, and time-of-flight mass analyzers coupled to GC. Covers scan, SIM, and MS/MS acquisition modes for identification and quantitation.

Lesson 1 • Ion-Exchange and Ion Chromatography

Describes strong and weak ion-exchange resins and suppressed conductivity detection for ionic analytes. Covers eluent selection and suppressor operation.

Lesson 2 • Mobile Phase Scouting and Optimization

Applies systematic scouting grids and triangle optimization to identify optimal mobile phase composition. Integrates pH, buffer type, and organic modifier into a structured workflow.

Lesson 3 • Normal Phase and HILIC Separations

Contrasts normal-phase adsorption with hydrophilic interaction chromatography for polar analytes. Addresses water layer formation and mobile phase water content in HILIC.

Lesson 4 • Particle Technology and Column Formats

Compares fully porous, superficially porous, and sub-2-micron particles for efficiency and backpressure. Introduces monolithic and core-shell column formats.

Lesson 1 • LC-MS Interface Technologies

Explains electrospray ionization, atmospheric pressure chemical ionization, and APPI interfaces. Covers ion suppression, matrix effects, and interface parameter optimization.

Lesson 2 • LC-MS/MS Quantitative Analysis

Applies multiple reaction monitoring and data-independent acquisition for trace quantitation in complex matrices. Covers internal standard selection and matrix-matched calibration.

Lesson 3 • Refractive Index and Evaporative Light Scattering

Describes RI and ELSD as universal detectors for non-UV-absorbing analytes such as sugars and lipids. Addresses gradient incompatibility and nebulization optimization.

Lesson 4 • Fluorescence and Electrochemical Detectors

Explains excitation-emission fluorescence detection and amperometric electrochemical detection for high-sensitivity applications. Covers derivatization strategies to extend detector applicability.

Lesson 1 • Design of Experiments in Chromatography

Applies factorial and response surface designs to optimize multiple method parameters simultaneously. Interprets interaction plots and contour maps to locate robust operating conditions.

Lesson 2 • Systematic Scouting and Screening

Applies column and mobile phase screening arrays to rapidly identify promising separation conditions. Uses gradient scouting runs to estimate retention and selectivity space.

Lesson 3 • Defining Method Requirements and Goals

Establishes analyte list, matrix complexity, detection limits, and throughput targets before column or mobile phase selection. Translates analytical goals into measurable method performance criteria.

Lesson 4 • Temperature and Flow Rate Optimization

Quantifies temperature effects on viscosity, diffusion, and selectivity in both LC and GC. Applies van Deemter optimization to set carrier gas or mobile phase flow rates.

Lesson 1 • System Suitability and Ongoing QC

Establishes system suitability criteria for plate count, tailing factor, resolution, and RSD before each analytical run. Integrates control charts and QC samples for ongoing performance monitoring.

Lesson 2 • Calibration Strategies and Curve Fitting

Compares external standard, internal standard, and standard addition calibration for different matrix types. Evaluates linear, quadratic, and weighted regression models for best fit.

Lesson 3 • Validation Parameters and Acceptance Criteria

Defines specificity, linearity, accuracy, precision, LOD, LOQ, range, and robustness as core validation parameters. Links each parameter to its regulatory acceptance criterion.

Lesson 4 • Robustness and Ruggedness Testing

Applies Plackett-Burman screening to identify method parameters most sensitive to small deliberate changes. Distinguishes robustness from ruggedness and documents acceptable operating ranges.