HPLC Introduction & Safety

P. van Poppel – Field Support Specialist March 2010

Resources

Solvents

Pump

Sampler

Sample

Column

Detector

Accessories (Fraction Collector, Oven, Valves…)

Data System

Available Pumps

Pumps

• Current– 210/218 Isocratic, one pump one solvent, combine

two to get a gradient system.– 212 LCMS pump of choice– SD1/SD2 Dual piston, pulseless, isocratic pump.

• Obsolete– 220 Isocratic, single piston pump.– 230 Ternary pump with proportioning valves, single

piston– 240 Quaternary pump, single piston

Available Pumps

• Interchangeable heads: 5/10/ 25/50/100/200• 0.1 to 200ml/min flow rate• Easy maintenance• Upto 8700psi rating• Add pumps for gradients!

210/218Analytical to Preparative

Pump

SD1 Preparative Pump

• Interchangeable 50, 200,500,800 mL/min pump heads• Pulse-free flow from 0.001 mL/min to 3200 mL/min

Available Samplers

Samplers

• Current– 410 Mid-size, single tray, full, partial and user program

– 420 Full-size, 8 segment interchangeable trays, full, partial and user program

– 430 Full size (optional robotic arm), 96-well plates full, partial and user program.

– Manual injector

• Obsolete– 400 Single tray, full loop injection

Note: Cooling options are available in 410, 420, 430

Available Samplers

420 Autosampler

• 1ul to 1ml capacity• Multi vial formats

Manual Injector 410 Autosampler

• 1ul to 10ml capability• 84 2ml vials• 24 10ml vials• Interchangeable Syringes

430 Autosampler

• 0.1ul to 1ml capacity• 96/384 well plates

Available Detectors

Detectors

• Current

– 325 Single/dual wavelength UV/VIS

– 335 Photodiode array - PDA

– 356 RI detector

– 380/385 Evaporative Light Scattering detector

Available Detectors

• 190-950nm wavelength range• Analytical - Preparative flowcells• Peak Purity• Spectral Calculator•“On the fly” spectra• Unlimited wav. extracts

335 Diode Array

• 190-900nm wavelength range • Analytical - Preparative flow cells• Benchmark Noise & Drift specs• Patented Dual Path cell• UV only option• Upgradeable from single-dual wavelength

325 UV-Vis 380/385 ELSD

• No chromaphore required• Universal response• Full gradient and solvent compatibility• No need for derivatisation• Excellent baseline stability• Complements other LC detectors• 10 degree evaporation• Patented Gas flow design• Programmable LED Intensity

356 RI

• High sensitivity for non-absorbing molecules•Superior peak shape•Full Automation via Galaxie™ •Low dead-volume flow-cell •Active temperature compensation •Long-Life LED

Available Detectors

Detectors

• Obsolete

– 340 single wavelength UV, D2 (Thermo)

– 350 RI detector

– 355 RI detector

– 363 Fluorescence (Hitachi D2 lamp)

– 370 Electro chemical (Antec)

– ELSD 2100 (/ice)

– Hitachi L-2485 FL

Available Accessories

Accessories

• Current– 440 Fraction Collector– 500 Column Valve Module– 510 Mistral oven, serial communication– 530 Column Valve Organizer – Various Rheodyne and Valco manual valves

• Obsolete– 701 Full size x-y movement fraction collector– 704 Half size x-y movement fraction collector– 520 Oven

Available Accessories

• 3 or 6 column selection• 2 independent heater zones• Diverter valves for column switching/back flushing and fraction collection• 6 solvent selection for method development• Utilised with Fusion AE for Automated Method development

500 Column Valve Module 530 Fluidics Module

• Utilised in tandem with LC ReSponder• 10 collects,1 waste,1 recycle valve• Blow fractions lines clear with N2• Configures multi-solvent inlets for pump• Pneumatically driven for safety• No moving parts

440 Fraction Collector

• Accurate, reproducible fraction collection• Efficient sample collection• Large fraction capacity• Huge flow rate range• Simple, intuitive software• Compact footprint• Optional Fume Extraction Kit