Transcript Redefining HPLC Purification - 1200 Series Purification

Redefining HPLC purification
Agilent 1200 Series Purification Platform
Christian Gotenfels
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
_
_____________________________
Delay calibration – how does it work?
____________
18
19
Mass-based fraction collection
20
Recovery strategies ___________________________________
21
Advantages
Agilent’s CAN network
22
Agilent’s integrated safety concept
23
Summary
24
2
Preparative HPLC in the pharmaceutical industry
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
3
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)
4
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
5
Agilent 1200 Series purification system (AS)
Purification systems (AS)
designed for:
• Purification of sample amounts
up to the low mg range
• for flow rates below 5 (10) mL/min
Pumps
- Isocratic
- Binary
- Quaternary
Injectors
- Standard
- Well-plate
- Preparative
- Thermostatting
- Manual
Column
holder
- Thermostatted
compartment
- Column organizer
• Minimum delay volumes for high
separation performance resulting
in high recovery and purity
- Valves, columns
and supplies
Detectors
- VWD
- MWD
- DAD
- MS
- UIB for third party detectors
Fraction
collectors
- Fraction collector (AS)
6
Agilent 1200 Series purification system (PS)
Pumps
- Preparative
- Gradient
- Extension
Injectors
- Preparative
- Thermostatting
- Manual
Column
holder
- Column organizer
Detectors
- MWD
- DAD
- MS
- UIB for third party detectors
Purification systems (PS)
designed for:
• Purification for sample amounts < 1 g
• Flow rates up to 100 mL/min
- Valves
- Columns and supplies
Fraction - Fraction collector (PS)
collectors
7
Agilent 1200 Series preparative pump
Preparative pumps deliver:
• Flow rates up to 100 mL/min at 400 bar
without changing pump heads
• Precise high pressure binary gradient formation
with low delay volume (700 µl) for preparative
and analytical applications down to 0.5 mL/min
• Automated seal wash using a peristaltic pump for
extended seal lifetime
• Pressure under and over sensing turns off the
pump if the column gets blocked or leakages
occurs
• Automated electro-magnetic purge valve
8
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
9
Agilent 1200 Series fraction collectors
Fraction collectors:
• 1200 Series fraction collector
Preparative Scale (PS)
• 1200 Series fraction collector
Analytical Scale (AS)
Preparative scale (PS)
< 100 mL/min
Analytical scale (AS)
< 10 mL/min
• 1200 Series micro fraction collector
Micro scale
< 100 µL/min
10
The reason for 3 dedicated fraction collectors
- Influence of peak dispersion
Dispersion:
2 
  r4  F  L
24 Dm
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
11
Agilent 1200 Series fraction collectors
- Accessories
Fraction collector accessories:
Well plate tray
• Well plates (96 and 384)
Small vials tray
• Vials and Eppendorf tubes
• Small or large tubes (up to 45 ml)
Tubes trays
• Customized racks
• Funnel trays for collection of large
fraction volumes
Customized rack
• Fraction collection can be increased
by using 3 fraction collectors in parallel
Multi-funnel trays
12
Agilent’s purification software solutions
Software solutions:
Advanced solution
Walk-up solution
21 CFR Part 11 compliance
with Security Pack
Easy management of
high samples numbers.
Flexible for
workflow integration
System management for
secure access,
Ideal for non-expert users
Agilent ChemStation
Software
Agilent Purification
Software
Agilent Easy Access
Software
Standard solution
Standard functionality for
easy system usage
Remote data browsing and purity reports at your desk
Agilent Data Browser
Dedicated software to match your purification needs
13
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
14
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
15
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
16
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
17
Agilent’s patented delay sensor
– automated calculation of the delay time
Fraction delay sensor:
• Comparable to a
simple detector in
the fraction collector
18
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
19
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
20
Recovery strategies
Peak
Peak
Waste
Waste
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
Fraction collector 1
Fraction collector 2
Column
Detector
Click here to see animation
Valve
Fraction collector 1
Recovery device
Fraction collector 2
21
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
22
Integrated safety concept
Health and safety:
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
23
Agilent 1200 Series purification platform
- key features
• Dedicated systems with low delay volumes
Optimized for high recovery and purity and desired throughput
• Software solutions for different user needs
Allows you to configure your own purification system
• Automated delay volume calibration
For easy and fast system set-up
• Solutions for recovery collection
Ensures that your valuable sample can never be lost
• Integrated safety concept
Safe leak handling/detection and forced fume extraction gives you walk-away confidence
• System Intelligence with CAN network
Data processing in real time for instantaneous and precise fraction collection
• Single-vendor solutions
For all your equipment, columns, supplies and support
24
For the latest information visit:
www.agilent.com/chem/purification
25