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Christian Gotenfels

Christian Gotenfels. Contents. Chapter Page The role of preparative HPLC in the pharmaceutical industry 3 Agilent’s portfolio 4 Hardware 5 - 12 Software 13 - 1 7 Techniques

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Christian Gotenfels

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  1. Christian Gotenfels

  2. Contents Chapter Page The role of preparative HPLC in the pharmaceutical industry 3 Agilent’s portfolio 4 Hardware 5 - 12 Software 13 - 17 Techniques Agilent’s patented delay sensor _ _____________________________ 18 Delay calibration – how does it work? ____________ 19 Mass-based fraction collection 20 Recovery strategies___________________________________ 21 Advantages Agilent’s CAN network 22 Agilent’s integrated safety concept 23 Summary 24

  3. Challenges Preparative HPLC in the pharmaceuticalindustry Challenges for the pharmaceutical industry: • Pharmaceutical companies are striving to develop faster and more efficient ways to generate new drug candidates against the increasing number of biological targets.In order to achieve this goal improvements are required in the following areas: • High throughput synthesis • High throughput purification • High throughput screening Preparative HPLC…for purification is fast enough to keep pace in drug discovery process

  4. Systems Agilent’s approach - dedicated systems Designed for various sample quantities, optimized for the high recovery and purity Analytical scale (AS) For flow rates < 5 (10) mL/min Agilent 1200 Series purification system (AS) Preparative scale (PS) For flow rates < 100 mL/min • Agilent 1200 Series purification system (PS)

  5. Systems Agilent 1200 Series purification systems • General system hardware features: • Modular approach for maximum flexibility • Compact, stackable design (typical bench space for MS-based fraction collection only 2 m) • Early Maintenance Feedback (EMF), a time-saving maintenance tool alerts when maintenance is due • Analytical and preparative columns with different selectivities for specific purification tasks • Integrated safety concept for walk away confidence

  6. Hardware Agilent 1200 Series purification system (AS) - Isocratic - Binary- Quaternary Pumps Purification systems (AS)designed for: - Standard- Well-plate- Preparative- Thermostatting- Manual Injectors • Purification of sample amounts up to the low mg range • for flow rates below 5 (10) mL/min • Minimum delay volumes for high separation performance resulting in high recovery and purity Column holder - Thermostatted compartment- Column organizer- Valves, columns and supplies Detectors • VWD- MWD- DAD- MS- UIB for third party detectors Fractioncollectors • Fraction collector (AS)

  7. Hardware Agilent 1200 Series purification system (PS) Pumps - Preparative- Gradient - Extension Injectors - Preparative- Thermostatting- Manual Purification systems (PS)designed for: Column holder - Column organizer- Valves- Columns and supplies • Purification for sample amounts < 1 g • Flow rates up to 100 mL/min Detectors • MWD- DAD- MS- UIB for third party detectors Fractioncollectors - Fraction collector (PS)

  8. Hardware Agilent 1200 Series preparative pump Preparative pumps deliver: • Flow rates up to 100 mL/min at 400 barwithout changing pump heads • Precise high pressure binary gradient formationwith low delay volume (700 µl) for preparativeand analytical applications down to 0.5 mL/min • Automated seal wash using a peristaltic pump forextended seal lifetime • Pressure under and over sensing turns off the pump if the column gets blocked or leakages occurs • Automated electro-magnetic purge valve

  9. Hardware Agilent 1200 Series dual loop autosampler PS Dual loop autosampler PS delivers: • Injection from a variety of sample containers • Dual fixed loop concept • High draw speed for fast injection of high sample volumes • Needle wash mechanism for lowest carry over

  10. Hardware Agilent 1200 Series fraction collectors Fraction collectors: Preparative scale (PS) < 100 mL/min • 1200 Series fraction collector Preparative Scale (PS) • 1200 Series fraction collector Analytical Scale (AS) • 1200 Series micro fraction collector Analytical scale (AS) < 10 mL/min Micro scale < 100 µL/min

  11. Techniques The reason for 3 dedicated fraction collectors- Influence of peak dispersion Dispersion: Derived from Aris, Taylor and Golay equation Dispersion (peak broadening) between detector and fraction collector results in poor recovery and purity. The selection of the appropriate flow inner diameter in the collectors delivers the best recovery and purity. Volumetric peak variance F: Flow rate Dm: Molecular diffusion coefficient L: Capillary length Click here to see animation Capillary inner diameter

  12. Hardware Agilent 1200 Series fraction collectors - Accessories Well plate tray Fraction collector accessories: • Well plates (96 and 384) • Vials and Eppendorf tubes • Small or large tubes (up to 45 ml) • Customized racks • Funnel trays for collection of large fraction volumes • Fraction collection can be increased by using 3 fraction collectors in parallel Small vials tray Tubes trays Customized rack Multi-funnel trays

  13. Software Agilent’s purification software solutions Software solutions: Walk-up solutionSystem management for secure access, Ideal for non-expert users Agilent Easy Access Software Standard solution Standard functionality for easy system usage21 CFR Part 11 compliancewith Security Pack Agilent ChemStation Software Advanced solutionEasy management of high samples numbers. Flexible for workflow integration Agilent Purification Software Remote data browsing and purity reports at your desk Agilent Data Browser Dedicated software to match your purification needs

  14. Software Standard solution – Agilent ChemStation Agilent ChemStation delivers: • Full system control via graphical user interface • Standard purification functionality for easy system usage • Sophisticated trigger options • Fraction preview helps to identify the right trigger parameter • Fraction task provides straightforward purification data review • ChemStation Security Pack supports 21 CFR Part 11 compliance

  15. Software Advanced solution – Agilent purification software • Additional to the standard software solution: • Color coded graphical user interface for easy sample setup and sample/fraction tracking • Sophisticated export/import functionality integrates the system into the workflow • Extended reporting capabilities • Navigation panel for fast data review

  16. Software Walk-up solution – Agilent Easy Access software Agilent Easy Access software delivers: • Flexible administrator tools to set up user access, queue tracking and project management • Simple sample submission and status checking • E-mail notification of results • Rapid confirmation of molecular weight and target ion presence

  17. Software Data browsing and purity reports – Agilent data browser Agilent data browser delivers: • Allows fast and efficient remote data review • Data file tree structure for easy navigation • Display of sample purity

  18. Techniques Agilent’s patented delay sensor – automated calculation of the delay time • Fraction delay sensor: • Comparable to a simple detector in the fraction collector

  19. Techniques Delay calibration – how does it work? • Delay calibration: • The time between peak detection and fraction collection is called ‘delay time’. This parameter must be known for correct fraction collection. The Agilent fraction delay sensor provides an automated and precise way to measure this time. • Whenever the main flow, or the make-up flow, is changed the system must be re-calibrated • The fraction delay sensor turns delay calibration into a simple, automated task Click here to see animation

  20. Techniques Active splitting – mass based fraction collection Active splitting provides: • A broad range of variable split ratios • for optimizing applications, from • 25:1 to 100,000:1 • The constant, accurate split ratios are unaffected by viscosity, temperature and tubing length • Minimal post-column delay volumes minimizes dispersion and delay times • Split ratios does not influence fraction delay times. This eliminates the need to recalibrate the system Click here to see animation

  21. Techniques Recovery strategies Peak Peak Waste Waste • Agilent offers three strategies to ensure • recovery of valuable compounds: • Recovery valve for up to 12 recovery locations • Third party fraction collectors can be controlled by an external contact board • Agilent fraction collectors with software-controlled funnel trays, for 40, 80 or 120 funnels Fraction collector 1 Fraction collector 2 Column Detector Click here to see animation Recovery device Valve Fraction collector 1 Fraction collector 2

  22. Advantages Controlled Area Network - System Intelligence • The design of the 1200 Series modules delivers: • Operation independent of PC • Real-time data processing • Fractions are triggered by inter-module communication without computer delay due to CPU usage or network traffic • Automatic tray-recognition prevents overflow of fractions Click here to see animation

  23. Advantages Integrated safety concept Health and safety: • Standard in all 1200 Series modules: • Leak detection • Safe leak handling • Pressure under sensing in pumpsProviding walk-away confidence • Forced fume extraction for the fraction collectorAllows system use outside a fume cupboard

  24. Summary Agilent 1200 Series purification platform - key features • Dedicated systems with low delay volumesOptimized for high recovery and purity and desired throughput • Software solutions for different user needsAllows you to configure your own purification system • Automated delay volume calibrationFor easy and fast system set-up • Solutions for recovery collectionEnsures that your valuable sample can never be lost • Integrated safety conceptSafe leak handling/detection and forced fume extraction gives you walk-away confidence • System Intelligence with CAN networkData processing in real time for instantaneous and precise fraction collection • Single-vendor solutionsFor all your equipment, columns, supplies and support

  25. For the latest information visit: www.agilent.com/chem/purification

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