hplc introduction & safety p. van poppel – field support specialistmarch 2010
TRANSCRIPT
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