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
Fast prep
Integ fc co;\llection
Perform
Consumables
Ana to skid

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
We have uncovered the market need -

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 Feature Advantage Benefit
Interchangeable Pump Heads
(5 to 200ml/min)
Unrivalled flexibility for any application
Economical and Convenient with Excellent Performance
Open access for ease of maintenance
Easy for the user to correct basic issues resulting in more uptime
Dependable
Simple and system Durability
Analytical to Prep on one System
One system will do the work of two providing high performance flexibility
Performance and Reliability at an Excellent Price/Performance ratio

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
Feature
Advantage
Benefit
Needle in Path
Design
No need for Injection loops
No Carryover
Versatility, Economical
Performance
One Injector, large or small inject volumes
Simplifies the system, no need to swap components for analytical to prep
Accuracy and Seamless Simplicity
One Autosampler, all rack formats
No need for multiple injector options
Versatility
Simplicity

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
Feature
Advantage
Benefit
NEW 4x0.25 Low Volume Cell
Optimised for Analytical to Prep, no need to change Cells
Performance and Versatility and Dependable
Tool Free maintenance
Easy to change lamps and flow cells
Simplicity
Range of cells for ul/min to Litres/min
Fits all applications from Analytical to Benchtop Prep
Versatility and Performance

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
Feature
Advantage
Benefit
Modern Design, improved movement and hardware
Significant improvement ergonomically on 701
Pleasing Appearance with Stylish fast Collector movement
Simplified user interface
Easy to use, does what you ask it to do both manually and automated
Simplicity and precious Dependability for customer fractions
Inject/Collect and Re-inject
Collect into A/sampler rack then re-insert into the Sampler for Fraction Purity Determination
Provides Confidence in Fraction Puritys without the need to do any liquid transfers

20. Varian 445-LC Scale-up Module
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

21. Varian 445-LC Scale Up Module( SUM) - FAB
Feature
Advantage
Benefit
2 Solvent Selection valves for solvent switching and flexibility
Can prime up 4 most popular solvents thus saving time on re-priming
Simplicity and Dependability
Analytical and Prep Columns Positions for Automated Scale Up or Switching
Seamless and simple transfer from an Analytical analysis to Prep
Simplicity and Versatility
Integrated Manual Valves which are dedicated for Analytical and Prep
Push button valve positioning with automatic return, eliminate operator error
Even Simpler to use, eliminates user error improving Confidence in results

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
pump
inject
detect
SUM FC
Modular
Isocratic
25ml
Opt any
Manual
UV/Vis NA
Base
Grad
Man inj
Dual UV
option
Productive 1
Opt 50
Auto 5ml +man
yes
Productive 2
PDA
Prep
100ml
Opt 200
manual
Dual UV/Vis

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 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

33. Instrument Condition Gradient 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 46

47. Questions??