Become A Master HPLC - Analysis Specialist Training

Operate the piston pump to move the mobile phase and inject samples through the system, delivering liquid to the column and generating detector peaks.

Learn to operate the HPLC pump by opening the valve counterclockwise, priming lines A and B, removing air bubbles with the purge valve, and adjusting flow rates for analysis.

Open the pump valve and start flow to purge air bubbles until the line is air-free, with zero bar pressure; then condition the system with mobile phase.

Explore how a red liquid-detection sensor prevents leaks and stops the pump to protect mobile phase flow to the column, with hands-on troubleshooting and reset steps.

Compare pump modules across brands, showing similar working principles yet different designs, with compact and modular layouts, confirming identical analysis results across devices.

Understand the auto sampler module in HPLC, including automatic injections, 100 μl or 200 μl loop options, and how temperature control and method validation ensure stable readings.

Understand how autosampler trays and numbered positions organize sample wheels, standards, and solvents, using amber wheels to protect solutions, with software confirming tray capacity for accurate injections.

Explore how the autosampler injection system works, contrasting a moving needle with a fixed-tray design and showing how a tray with 25 wheels across five trays enables 125 samples.

Operate the column oven to set and monitor temperatures, observe how higher temperatures affect pressure and flow, and understand leak sensors that protect the HPLC system.

Learn practical techniques for washing an HPLC system, including cleaning the pump and column, using an intermediate connection, and applying proper flow rates to prevent contamination and preserve column integrity.

Understand the HPLC columns, including brands, sizes, diameters, and filling materials, and how specifications like C18, 3 micrometers, 4.6 x 100 millimeters affect retention time and traceability.

Learn to assemble the HPLC column by aligning the ends, protruding the line slightly, and tightening the nut to secure the seal and prevent sensor warnings.

Explore detectors used in HPLC systems, including UV-visible, fluorescence, and refractive index detectors, and learn how UV lamps and fixed versus scanning wavelengths determine absorption peaks and signals.

Learn how hplc software supports online analysis and offline data processing, set up methods for isocratic and gradient runs, configure detectors, wavelengths, and autosampler and column conditions.

Define the reference standard and prepare the sample for HPLC analysis, then formulate mobile phase and calibration solutions to quantify four components using retention time.

Plan the mobile phase volume for washing, conditioning, and analysis, using about 2300 ml as a practical guide, and include injector wash and auto purge to keep the system ready.

Learn to prepare a mobile phase by weighing and dissolving reagents in ultrapure water, creating a 71/29 buffer–methanol mix, and ensuring complete dissolution and homogeneity.

Learn to clean the HPLC system after analysis by washing lines, switch between reverse and normal phase, use isopropyl alcohol for polar residues, and maintain GMP logbooks for traceability.

Prepare and clean HPLC system before connecting mobile phase, using a two-stage wash to remove residues and flush lines A–D and injector washing line with methanol and water.

Learn how to wash and clean the HPLC system, assemble and connect a column with proper interconnects, prevent leaks, and store the column using manufacturer guidelines and specifications.

Learn how column impurities and mobile phase changes elevate pressure in HPLC, and use usage records to diagnose and correct pressure discrepancies.

Open the device modules and log into the software to enable real-time online analysis. Run auto purge to clean lines, including the autosampler injector line, with mobile phase and acetonitrile–water washes.

Clean the pump module by rinsing with pure water to remove salts and prevent residue, monitor for fluid leaks, and resume flow only after drying the sensor area.

Learn how the hplc pump module starts flow in isocratic mode, linking interconnection and column effects to pressure, with methanol, water, and acetonitrile altering flow and pressure.

Connect the mobile phase and washing solutions to the device; perform two-stage washing with a high-water solution followed by a high-organic solution, then condition and connect the column.

Prepare and analyze four active ingredients using a validated HPLC method, building calibration curves from three-point standards and ensuring ppm-appropriate concentrations with fully dissolved samples.

Learn to prepare and weigh reference solutions for HPLC analysis, using vitamin C and B3 standards, solvent as water, ultrasonic dissolution, and careful weighing to ensure accurate results.

Weigh stock standards, dissolve, and transfer to 100 ml volumetric flasks to prepare 2000 ppm stock solutions for HPLC analysis. Use separate spatulas, record each weight, and avoid contamination.

Weigh a ten milliliter sample, ensure homogeneity, dissolve in the standard solvent using density calculations, then fill to the volume line and dilute to 100 ml for four components.

Prepare stock standards by mixing vitamin C, B1, B3, and B6, use an ultrasonic bath to dissolve, then dilute to three calibration levels for accurate HPLC.

Prepare and filter standards and samples for HPLC by using 0.45 micron RC filters, loading amber wheels, and ensuring proper injector setup before analysis.

Place standards and samples on the autosampler tray according to a batch table, mix standards by concentration, and record positions for vitamin C and recombinants to ensure correct timing.

Learn to create and save a new HPLC method in real-time software, configure modules, detectors, autosampler, pump, and gradient or isocratic settings, and set analysis and purge parameters.

Create and configure a batch table for an HPLC analysis, assign samples to autosampler positions and trays, select methods, set injection volumes, and define data file names for automated reporting.

Condition the device and column by running the mobile phase to establish a stable baseline, monitor pressure and detector settings, and ensure smooth peaks before analysis.

Initiate hplc analysis by starting the batch, monitor injections and retention times, assess peak symmetry, replace the column if needed, and perform data integration with calibration standards.

Configure a shutdown wash for two lines (lines C and D) with 15-minute washes, stop the pump and detector, and turn off the degasser in the batch table.

Learn to determine retention times of reference standards using automatic integration in chromatography software, including wizard-guided parameter setup, drift adjustments, and manual vs automatic integration practices.

Explore manual and automatic integration in HPLC, adjust slope, drift, and minimum area settings, and learn to edit, delete, and simulate peaks for a single reference standard like Vitamin C.

Discover how to quantify vitamin C by hplc using external standards, select retention time, integrate peak areas, and build a three-point linear calibration curve with proper units.

Determine retention times and peaks for vitamins B1, B3, and B6 in an HPLC analysis, assess impurities using standards, injections, and solvent checks for accurate identification.