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Varian 940-LC Analytical to Prep HPLC

Varian 940-LC Analytical to Prep HPLC. Varian 940-LC Silver Bullets. The 940-LC one instrument – many uses: High performance automated Analytical HPLC High performance semi-preparative HPLC High performance prep LC. Varian 940-LC Silver Bullets. Flexible and capable

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Varian 940-LC Analytical to Prep HPLC

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  1. Varian 940-LC Analytical to Prep HPLC

  2. Varian 940-LC Silver Bullets • The 940-LC one instrument – many uses: • High performance automated Analytical HPLC • High performance semi-preparative HPLC • High performance prep LC

  3. Varian 940-LC Silver Bullets • Flexible and capable • Wide flow rate from 0.1 ml/min to 200 ml/min • Dual path-length flowcells for extended Abs • One Injector, analytical or prep volumes (5ul to 4.5ml) • Superb fraction collecting • Galaxie real-time peak detection • Practical collect and inject • Much Easier interface, it does what you tell it! • Cleaner UI

  4. Goals of the 940-LC Analytical to Prep • Aim • Become market leader in this $180M segment • Double market share • Market • Continue to focus on Pharma / BioPharma • Academic • Strategy • Deliver innovation in hardware, consumables, software and services

  5. Varian 940-LC Instrument Overview • Factory tested, pre-plumbed instrument: • Plug and play operation • High performance semi-preparative HPLC • Gradient Analytical HPLC • Autoprep System • Choice of UV or PDA detector • Dual path flow cell for unmatched range and scalability • No compromise performance for prep and analytical – switch with a key stroke – no plumbing or hardware changes needed! • The most flexible analytical to prep HPLC available

  6. Varian 940-LC Instrument Overview • Scale-up Module for automated methods transfer from analytical to semi-prep, or switching from Analytical to Prep work • Versatile and robust solvent-delivery system • New loopless autosampler – wide volume range • NEW Fraction collection with • Complete range of collection modes such as slope, threshold and time slicing • Wide range of vessels, including autosampler rack • Comprehensive range of columns and consumables • More productivity via GalaxieTM software • Valve automation for genuine unattended operation • NEW direct control interface

  7. Varian 940-LC Versatility and Performance • Easy to change pump heads • Versatile solvent delivery systems • Powerful pumps allow 4000psi at 25mL/min operation for fast chromatography • Ideal for 4.6mm and 1” column • Able to use smaller 3 & 5 um particles columns at elevated pressures • Improves productivity • Save solvents and waste costs • Short run times save time • High resolution separations = high purity 1ml/min Analytical 21ml/min Prep

  8. Varian 940-LC Pumps • Same range of accessories as PS210/218 • 5ml Head to 200ml Head • Easily interchangeable pressure modules thru removable side panel. • Easy access to mixer • Self maintenance and versatility features have been completely retained with no compromises – integrated systems are notoriously difficult to maintain so this is Unique

  9. Varian 940-LC Pumps - FAB

  10. Varian 940-LC Autosampler • Prep version of the 920-LC Sampler • 5ul to 4.5ml Injection range • No necessity to combine a Manual injector • No need to dedicate injector to prep or analytical • One Sampler does all! • Cooled version • Loopless Design • Needle in path Design • No carryover • 2008 Technology – no changing loops

  11. Varian 940-LC Autosampler - FAB

  12. Varian 940-LC UV/Vis - PDA Detectors • High Sensitivity for Analytical applications • Extended absorbance – desensitised, for Prep Applications • All in One Cell! • NEW 4 x 0.25mm Cell low dead volume • Excellent performance at low flow rates for Analytical • Excellent Extended Absorbance for Prep

  13. Varian 940-LC Dual Pathlength Flowcell • Absorbance Detector • Extended absorbance range without re-plumbing (<70 AU) • NEW 4mm x 0.25 mm • 4mm x 0.15 SuperPrep • Low dispersion • UV/Vis Detector • PDA Detector • Low noise ~ +10 mAU • Low Drift ~ 1 mAU/hr

  14. Agilent 1200 Flowcells

  15. Varian 940-LC UV/Vis - PDA Detectors - FAB

  16. Varian 440-LC Fraction Collector • Fully controlled by software for foolproof collection by slope, level or timeslice • Known fluid path for accurate fraction collection • Software display of collected fractions and source injections • Reliable operation • Compatible with a wide range of containers • Belart and other optional racks • Microvial to 50ml tube capacity • Autosampler rack compatibility provides practical collect and re-inject capabilities

  17. Varian 440-LC Fraction Collector • We chose not to collect Peak 3 • Peaks 1,2,4,5,6,7 collect with exceptional accuracy • Excellent algorithm, and simple delay volume calculation • Slope/Threshold based on real numbers!!

  18. Varian 440-LC Fraction Collector • Fraction Log shows peak location using colour coded rack assignment

  19. Varian 440-LC Fraction Collector - FAB

  20. Optimized flow paths for automated scale-up Separate optimised flow paths for Prep and Analytical columns Solvent selection valves for 4 solvents ( 2 per pump) Integrated priming valves for easy solvent changes Two Integrated Push Button Manual valves for Analytical and prep Injections Varian 445-LC Scale-up Module

  21. Varian 445-LC Scale Up Module( SUM) - FAB

  22. Varian 940-LC Software Usability • Software designed for Prep users • Enhanced manual control • One page status and chromatogram • Real time fraction collection display • Advanced peak sensing for better fraction collection • Purity displays for absorbance detection – standard! • Documentation and audit trails for traceability and regulatory needs

  23. Varian 940-LC Configurations

  24. 940-LC Pricing • Market driven pricing • Similar pricing to existing systems at the same level • Opportunity to move system price up with valuable options • General move to higher ASP systems

  25. Varian 940-LC Consumables • Columns designed for Prep users • From 3 to 10mm media • Wide range of C4 to C18 • Silica and Polymer media • From 2mm to 2” ID • Unique solutions for dedicated prep with load and lock systems • Matched Analytical to Prep columns and packing's to assure smooth scale up • Solutions for hardware, software and consumables

  26. Applications of the 940-LC

  27. Using Linear Scale-up Principles and the Varian 940-LC with Scale-Up Module for the Isolation of 5-Hydroxytryptophan from Griffonia Simplicifolia Dennis Hoobin Varian Australia Pty Ltd 679 Springvale Rd Mulgrave VIC Australia

  28. Varian 940-LC Analytical to Prep Instrument

  29. Introduction • Griffonia simplicifolia is a tall bushy vine found in Ghana, Nigeria, Liberia, Ivory Coast, Cameroon and Congo • The vine produces pods containing 3 large legumes • The legumes are a rich source of 5-hydroxytryptophan (5HTP) • 5-hydroxytryptophan produced by the human body from the essential amino acid L-hydroxytryptophan (LT) • 5-HTP has been clinically used to increase serotonin production to combat many mental illnesses like depression and schizophrenia among others

  30. Instrumentation • Varian 940-LC Semi-Prep HPLC (part no 02102201-00) • Binary gradient pumps • 128 position autosampler with 5mL syringe • Scale Up Module to allow seamless scalability from Analytical to Semi-preparative applications • Optimised Dual path flowcell • Diode Array detector • Accurate Fraction collection • Columns - Varian XRs C18 • Analytical column -150 mm, 4.6 mm ID, 5u • Preparative column - 150 mm, 21.2 mm ID, 5u

  31. Instrument Features • Scale Up Module • Fully Automated unit allowing switching between analytical and preparative columns • software-controlled module switches from analytical to preparative operation to ensure that chromatograms are correlated • Flow cell • optimized dual path flowcell that spans analytical to prep flow rates (1–25 mL/min) • the dual path flowcell design provides the ability to work over an extended absorbance range (mAu to tens of Au)

  32. Instrument Conditions • Scaling Up • Scaleable System Elements Absolute System Elements • Flow Rate Packing Material • Tubing Area Column Length • Sample Injection Volume Sample concentration • Solvent Consumption Gradient Slope • Run Time • Back Pressure • Flow Rate Packing Material

  33. Instrument ConditionGradient conditions • Mobile phase 85% A (H2O) 15% B (MeOH) • Flow rates: • 1 mL/min (analytical) • 21.2 mL/min (preparative) • Injection volume 50 μL analytical–4 mL preparative • Dual Array PDA detector: monitoring 2 channels at 230 and 280 nm.

  34. Analytical to Preparative 50 μL analytical injection of 5HTP standard

  35. Diode Array Chromatogram Diode array display. Quadrant 1: Chromatogram. Quadrant 2: 3D diode array data. Quadrant 3: Isogram. Quadrant 4: UV spectrum derived from chromatogram at cursor position

  36. Instrument Scale up Chromatograms 50 μL analytical injection of legume extract 4 mL preparative injection of legume extract, full scale. Note the extended range absorbance maximum of 8.5 abs

  37. Purity Check • Fraction between 3 and 3.5 min in the preparative chromatogram was collected resulting in a fraction volume of 10.1 mL • Reinjection of the collected fraction onto the Analytical column results in a single peak chromatogram. Overlayed with the PDA peak purity trace, the flat top of the purity trace indicates a pure compound

  38. Discussion • A good estimate of the amount of the % of HTP in dry Griffonia seeds can be made • The entire 10 mL fraction contains 17.2 mg • 1gm of seeds contains 215mg of 5HTP • Approx. 20% dry weight • G. Simplicifolia seeds are a very rich source of 5HTP

  39. Conclusion • Varian 940-LC is a powerful automated system to allow easy development of HPLC preparative separations • Developing an analytical method – seamless scale up to an optimised preparative columns • Excellent correlations between analytical and preparative chromatograms • Confidence in the purity of fractionated material

  40. Varian 940-LC Summary The 940-LC one instrument – many uses • High performance automated Analytical HPLC • High performance semi-preparative HPLC • Versatility • Pumping system able to operate at analytical flow rates and scalable up to prep flows • Dual path-length flow cell technology for unmatched range • Scale-up Module for automated methods transfer • Advanced fraction collection with full software control • Complete range of columns and consumables • Prep enhanced Galaxie TM software and valve automation for genuine unattended operation

  41. Status of Varian HPLC at Release • Installation/setup  • Perceptions  • Ease of ordering  • Operational flexibility  • Competitive performance  • Competitive specifications  • Ease of use  • Reliability  • Productivity  • Validation and Compliance  • Flexibility/expansibility (especially in semi-prep)  • Competitive pricing  • Wide product range – hardware, columns, consumables 

  42. What About the Modular HPLC?

  43. Modules Overview • What about modules? • Low price manual, isocratic systems • After market module business • Odd configurations/applications not served by iHPLC • Retaining • 210/218, SD-1 and 212 pumps • 325/335/356 detectors • CVM 500 • Still available but no need to actively sell due to iHPLC • 420 for Online SPE • 410/430 Yet, they will come! Price Increases so beware!

  44. Modules Overview • Obsoleting • 220/230/240 ternary pump module • 340 UV detector • 355 RI detector • 363 Fluorescence detector • 400 • CVO module • 701 fraction collector

  45. Varian 356-LC RI Detector

  46. Conclusion for 900 Series Compact and Stylish Integrated Systems Retain flexibility and maintenance features Top Tier hardware performance Top Tier User Interface – pleasant experience Simplicity in a box – Install , Use and Maintain 1st Truly Integrated HPLC’s on the Market 46

  47. Questions??

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