redefining hplc purification agilent 1200 series purification platform christian gotenfels
TRANSCRIPT
2
Contents
Chapter Page
The role of preparative HPLC in the pharmaceutical industry3
Agilent’s portfolio4
Hardware5 - 12
Software13 - 17
Techniques
Agilent’s patented delay sensor _ _____________________________18
Delay calibration – how does it work? ____________19
Mass-based fraction collection20
Recovery strategies ___________________________________21
Advantages
Agilent’s CAN network22
Agilent’s integrated safety concept23
Summary 24
3
Preparative HPLC…for purification is fast enough to keep pace in drug discovery process
Preparative HPLC in the pharmaceutical industry Challe
nges
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
Challenges for the pharmaceutical industry:
4
Agilent’s approach - dedicated systems
Agilent 1200 Series purification system (PS)
Analytical scale (AS)
Preparative scale (PS)
Syste
ms
Agilent 1200 Series purification system (AS)
For flow rates < 5 (10) mL/min
For flow rates < 100 mL/min
Designed for various sample quantities, optimized for the high recovery and purity
5
Agilent 1200 Series purification systemsSyste
ms
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
Agilent 1200 Series purification system (AS)
• 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
- Isocratic - Binary- Quaternary
- Standard- Well-plate- Preparative- Thermostatting- Manual
- VWD- MWD- DAD- MS- UIB for third party detectors
Pumps
Injectors
- Thermostatted compartment- Column organizer
- Valves, columns and supplies
Column holder
Detectors
Fractioncollectors
Hardw
are
Purification systems (AS)designed for:
- Fraction collector (AS)
7
• Purification for sample amounts < 1 g
• Flow rates up to 100 mL/min
Agilent 1200 Series purification system (PS)
- Preparative- Gradient - Extension
- Preparative- Thermostatting- Manual
- Fraction collector (PS)
Pumps
Injectors
Column holder
Detectors
Fractioncollectors
- Column organizer
- Valves- Columns and supplies
- MWD- DAD- MS- UIB for third party detectors
Hardw
are
Purification systems (PS)designed for:
8
• 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
Agilent 1200 Series preparative pumpHardw
are
Preparative pumps deliver:
9
• 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
Agilent 1200 Series dual loop autosampler PSHardw
are
Dual loop autosampler PS delivers:
10
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
Agilent 1200 Series fraction collectors
Analytical scale (AS) < 10 mL/min
Micro scale < 100 µL/min
Hardw
are
Fraction collectors:
11
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.
mD
LFr
24
42
Volumetric peak varianceF: Flow rateDm: Molecular diffusion coefficientL: Capillary length
The reason for 3 dedicated fraction collectors- Influence of peak dispersion
Techn
iques
Derived from Aris, Taylor and Golay equation
Click here to see animation
Dispersion:
Capillary inner diameter
12
• 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
Agilent 1200 Series fraction collectors
- Accessories
Well plate tray
Multi-funnel trays
Tubes trays
Small vials tray
Customized rack
Hardw
are
Fraction collector accessories:
13
Dedicated software to match your purification needs
Agilent’s purification software solutions
Advanced solution
Easy management of high samples numbers.
Flexible for workflow integration
Agilent Purification Software
Standard solution
Standard functionality for easy system usage
21 CFR Part 11 compliancewith Security Pack
Agilent ChemStation
Software
Walk-up solution
System management for secure access,
Ideal for non-expert users
Agilent Easy Access Software
Remote data browsing and purity reports at your desk
Agilent Data Browser
Softw
are
Software solutions:
14
• 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
Standard solution
– Agilent ChemStation
Softw
are
Agilent ChemStation delivers:
15
Advanced solution
– Agilent purification software
Softw
are
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
• 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
Walk-up solution
– Agilent Easy Access software
Softw
are
Agilent Easy Access software delivers:
17
• Allows fast and efficient remote data review
• Data file tree structure for easy navigation
• Display of sample purity
Data browsing and purity reports
– Agilent data browser
Softw
are
Agilent data browser delivers:
18
Agilent’s patented delay sensor
– automated calculation of the delay time
Techn
iques
Fraction delay sensor:
• Comparable to a simple detector in the fraction collector
19
Delay calibration
– how does it work?
Click here to see animation
Techn
iques
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
20
Active splitting
– mass based fraction collection
Techn
iques
• 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
Active splitting provides:
Click here to see animation
21
Agilent offers three strategies to ensure recovery of valuable compounds:
1. Recovery valve for up to 12 recovery locations
2. Third party fraction collectors can be controlled by an external contact board
3. Agilent fraction collectors with software-controlled funnel trays, for 40, 80 or 120 funnels
Recovery strategies
Recovery device
Techn
iques
Fraction collector 1 Fraction collector 2
Peak Peak
Waste Waste
Fraction collector 1 Fraction collector 2
Detector
Valve
Column
Click here to see animation
22
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
Advan
tage
s
Click here to see animation
23
Standard in all 1200 Series modules:
• Leak detection
• Safe leak handling
• Pressure under sensing in pumps Providing walk-away confidence
• Forced fume extraction for the fraction collector
Allows system use outside a fume cupboard
Integrated safety conceptAdvan
tage
s
Health and safety:
24
• 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
Agilent 1200 Series purification platform
- key features
Summ
ary