Download Report

W h a t is a n ACQUI TY UPLC® System H a r dw a r e a n d D e t e ct ion Con side r a t ion s
Tore Caesar, August 21
© 2008 Waters Corporation
H PLC / UPLC Tu bin g
@75 Times
@75 Times
HPLC
@500 Times
UPLC
@1000 Times
© 2008 Waters Corporation
2
Sw a ge lok fit t in gs u se d for Acqu it y
Swagelok fitting
SS capillary
Gold plated stainless steel fitting
Two piece SS ferrule
© 2008 Waters Corporation
3
ACQUI TY UPLCTM
Binary Solvent Manager
Low volume flow path
Pressure limit set to 15,000 psi
(1000 bar)
Serial/parallel Flow Path
Four solvent choices A1, A2, B1, B2
UPLC pressure capabilities
— Materials
— S/W algorithms
User diagnostics
© 2008 Waters Corporation
4
Bin a r y Solve n t M a n a ge r – Ea sy Acce ss
“B”
Pump
Primary
“A”
Pump
Primary
“A” Pump
Accumulator
“B” Pump
Accumula
tor
“A” Pump
Solvent
Select Valve
Six Channel
Degasser
“B” Pump
Solvent
Select
Valve
Filter/ Mixer/ Tee
Assembly
© 2008 Waters Corporation
5
W a t e r s I n t e llige n t I n t a k e Va lve
How does t he I nt elligent I nt ake Valve work?
— An applied current generates a force, timed with the motion of
the pump plungers, that pushes a ball off its seat. In the case
of traditional passive check valves the ball is lifted off the seat
by the force of the solvent flow
Provides:
— More robust operation
© 2008 Waters Corporation
6
ACQUI TY UPLCTM
Sample Manager
Com ple t e ly n e w in j e ct ion pr oce ss
3 different injection modes depending on preference
4 to 40º C
2 plate sample compartment
2 wash solvents, low sample carry over
Fast cycle time
— 25 sec no wash, <60 sec dual wash
— Inject Ahead
Needle- in- Needle Sampling
— Puncture needle and sample needle
Optional Sample Organizer
0.1 – 50µL injection range
© 2008 Waters Corporation
7
N e e dle - in - N e e dle Sa m plin g Pr obe
Z’
1. Locate sample vial
2. Puncture the cover
Z
3. Lower sampling probe
and aspirate sample
© 2008 Waters Corporation
8
ACQUI TY UPLCTM
Sample Organizer
Supports high throughput/high capacity
Tem perat ure cont rolled ( 4 t o 40º C)
Expands capacity to either:
— 22 microtiter plates
o 8,448 sample wells
— 15 mid height plates
— 8 deep well plates
o 388 2- ml vials
o 196 4- ml vials
Access to any sample location
Small footprint – 10.5 inches (27cm)
© 2008 Waters Corporation
9
ACQUI TY UPLCTM
Column Heaters
Coumn heater
— Room Temp - 90°C
Pivot positioning
— Stacked mode
o with optical detector
— Swung out mode
o with MS detector
Flow path distance optimized
— Minimizes dispersion
— Column may be accessed from
either side
Swung out mode to interface with MS
© 2008 Waters Corporation
10
e Cor d ™ Te ch n ology
Paperless tracking of column history
Holder, tether and chip permanently
attached to column
Microchip encased by 16mm stainless
steel can
Nonvolatile read/write memory
eCord™ reader
— Fixed column manufacturing data
- Unique column identification
- Certificate of Analysis
- QC test data
— Variable column usage data
- Column use data
- Gathered through life of column
Tether
Holder
Encased
16mm
Microchip
© 2008 Waters Corporation
11
e Cor d ™ Te ch n ology
Waters quality control results
Paperless Certificate of Analysis & Performance Chromatogram Data
© 2008 Waters Corporation
12
e Cor d ™ Te ch n ology
User history file
© 2008 Waters Corporation
13
ACQUITY UPLC TM Column Manager
and Column Heater/Cooler
Column Manager
Thermostatted from 10 - 90°C
Automated switching among
four columns and a bypass
channel
eCord Information
Management for each column
Accepts column sizes from
2.1x 30mm to 4.6 x 150mm
Passive pre- column solvent
heating and post- column
cooling
Column Heater/Cooler
Same as Column Manager but
with no switching valves
© 2008 Waters Corporation
14
Colu m n Com pa r t m e n t
Passive pre-column solvent
heating and post- column
solvent cooling capabilities
are built-in.
© 2008 Waters Corporation
15
Colu m n M a n a ge r Flu idic Sch e m a t ic
© 2008 Waters Corporation
16
Up t o t h e M in u t e Colu m n Usa ge
I n for m a t ion
© 2008 Waters Corporation
17
Com ple t e Colu m n H ist or y
© 2008 Waters Corporation
18
ACQUI TY UPLCTM
Column Heater 30 cm
Thermostatted from 4 – 65 degrees
© 2008 Waters Corporation
19
UPLC™ D e t e ct ion
Technology challenges
Narrow peaks require:
— Higher data rates
— Faster digital filter time constants
Smaller peaks require maximum S/N
— High light throughput/transmission
UPLC™ separations require low
dispersion flow cells
— Maintain peak shape
— Lower cell volume
— Without generating high backpressures
© 2008 Waters Corporation
20
Re solu t ion / Low D ispe r sion
Flow Ce ll
Smaller cell volume – how?
Shorter pathlength
— Decreased sensitivity
A smaller cross- sectional area
— Reduces the amount of light transmission and baseline noise
increases
D
d
© 2008 Waters Corporation
21
Ligh t Gu idin g Flow Ce lls
How they work
A light guiding flow cell is an
optical fiber whose core
material is a fluid
Light
Path
The optical fiber consists of a
core material surrounded by a
cladding layer (Teflon AF).
— Light is guided through the
core by the process of total
internal reflection
— Light rays that encounter the
interface between the core and
cladding are reflected back
into the core with essentially
100% efficiency
TeflonAF
α
Mobile
Phase
TeflonAF
10mm or 25mm
© 2008 Waters Corporation
22
ACQUI TY UV / PD A
Flow Cell Options
High Sensitivity
25 mm Pathlength
2400 nL Volume
Analytical
10 mm Pathlength
500 nL Volume
© 2008 Waters Corporation
23
Flow Ce ll Com pa r ison Se n sit ivit y
Analyti
cal
0.20
High
Sensitivity
0.24
0.22
Delta
0.20
0.18
Area
140955
326881
2.319
93942
211138
2.248
0.16
0.14
Height
AU
0.12
0.18
0.10
0.08
0.16
Peak Width at
13.4%
0.06
0.04049
7
0.04
0.042033
1.038
0.02
0.00
-0.02
-0.04
0.14
-0.06
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
Minutes
AU
0.12
0.10
0.08
0.06
0.04
0.02
0.00
4.40
4.45
4.50
4.55
4.60
4.65
4.70
4.75
4.80
4.85
4.90
Minutes
© 2008 Waters Corporation
24
ACQUI TY UPLC™ D e t e ct or s
Optical Detectors
Four opt ical ACQUI TY UPLC Det ect ors
— Holistic design
o High acquisition data rates
o Low dispersion, small flow cells
Photo Diode Array
o Unmatched sensitivity
o Extended linear range (UV detectors)
Evaporative light
scattering
Tunable UV
Fluorescence
More HPLC methods can be transferred to gain UPLC benefits
More UPLC methods can be developed
© 2008 Waters Corporation
25
ACQUI TY UPLC™ D e t e ct or s
Mass Spectrometers
ACQUITY SQD
featuring the
SQ Detector
ACQUITY TQD
featuring the
TQ Detector
Single Quadrupole MS
Introduced Pittcon 2006
Tandem Quadrupole MS
Introduced ASMS 2006
© 2008 Waters Corporation
26
UPLC a n d M S
UPLC pr ovide s t h e follow in g be n e fit s for M S:
— Improved resolution for complex mixture analysis
— Narrower chrom at ographic peaks effect ively increase
concentration of analytes entering the MS source – increasing
signal intensity and improving MRM detection limits
— Shorter analytical run times without compromising
chromatographic resolution – increasing sample throughput
— I m proved resolut ion can reduce MS ion suppression by
separating species that co- elute in conventional HPLC –
improved MRM detection limits
© 2008 Waters Corporation
27
UPLC- M S/ M S vs. H PLC- M S/ M S
QM_HPLC_ESI+_4Compound_041205_010
MRM of 4 Channels ES+
309 > 280.84
1.06e5
Height
%
ESI+, Alprazolam
HPLC- MS/ MS
0.99
28719
0
0.20
0.40
0.60
0.80
1.00
QM_UPLC_ESI+_2Compound_042605_010
1.20
1.40
1.60
1.80
2.00
2.20
2.40
0.54
105629
2.60
2.80
3.00
MRM of 2 Channels ES+
309 > 280.84
1.06e5
Height
%
UPLC- MS/ MS
0
0.20
0.40
0.60
0.80
1.00
1.20
1.40
1.60
1.80
2.00
2.20
2.40
2.60
2.80
Time
3.00
Peak height increase 3.7 times.
Signal to noise ratio 4.7 times increase
© 2008 Waters Corporation
28
UPLC com pa t ibilit y h igh e r da t a r a t e
100 ms Dwell Time, 10 ms Delay
0.63
0.63
Convert the x
axis to scan
number
Peak Width = 1.8 s
Points Across Peak = 7
%
%
Time
0
0.25
0.75
1.25
0
100
1.75
105
110
115
120
125
130
Scan
5 ms Dwell Time, 5 ms Delay
0.63
0.63
Convert the x
axis to scan
number
Points Across Peak = 60
%
%
0
0.25
0.75
1.25
1.75
Time
0
1100
1200
1300
1400
Scan
© 2008 Waters Corporation
29
ACQUI TY Syst e m Con sole
Designed to simplify user interaction with the system, enabling many actions to be automated
© 2008 Waters Corporation
30
ACQUI TY UPLC™ Con sole
Access to status and control
© 2008 Waters Corporation
31
ACQUI TY UPLC™ Soft w a r e
Required features incorporated
in current software
— Empower™
— MassLynx™
— Stand Alone Console
— 3rd party MS software,
o Analyst, XCalibur, Hystar
More information
— System monitoring
— Performance history
— eCord column tracking
Easy to use
— Reduce training
© 2008 Waters Corporation
32
ACQUITY UPLC™ Technology
4 Years in the Making – 4 Years in the Market
• UPLC™ technology was introduced after 4 years
of focused research and development at Waters
and with collaboration with world renown LC
scientists in academia and industry
• Introduced the first holistically designed UPLC™
system at Pitt Con 2004
• Many new detectors and other UPLC™ enabled
components and software as well as many new
columns have been added from 2004 to 2008
• Respected scientists and experts have published
reports in peer reviewed journals about the
extraordinary results not possible before UPLC™
technology was commercially available
• Thousands of companies have made major
investments to increase revenue, reduce costs and
maximize their resources based on the proven
results from UPLC™
© 2008 Waters Corporation
33
Re a l - Tim e Syst e m M on it or in g a n d
Ale r t N ot ifica t ion
Waters Connections®
INSIGHT™ Enterprise
Server
Customer
Connections® INSIGHT™ Firewall
Service Agent
Waters
Firewall
Waters
Technical
Services
Waters
Field Service
Engineers
Connections® INSIGHT™
Service Agent
The Customer
One-way Communication
© 2008 Waters Corporation
34
Connections® INSIGHT™
E- m a il Cu st om e r Re por t Ex a m ple
System Performance Status Report
— Automatic monthly notification
service to be delivered to customers
via email
— Service is designed to keep
Connections INSIGHT customers
informed about the performance of
their ACQUITY UPLC™ systems
— Report includes:
o Instrument Counters, such: Auto
Sampler Injections, Lamp Hours,
Volume of Solvent Delivered,
Lamp Ignition
o Column Usage – Injections Per
Column
o Estimated PM visit
© 2008 Waters Corporation
35
Be n e fit s of Re m ot e M on it or in g
a n d D ia gn ost ics
Maxim ize your syst em upt im e and product ivit y t hrough proact ive
maintenance scheduling
Improve MTTR (Mean Time to Repair) via automatic notification
of a problem
Real- time Remote Troubleshooting for real- time Problem
Resolutions
— Allows Waters to remotely resolve system problems without the need
of an on- site engineer to minimize instrument downtime
Gain confidence in the accuracy and quality of your results
— Waters partners with you in managing the quality of your results
Predict potential problems by monitoring component and
consumable usage
— Waters provides reports that estimate timing of performance
maintenance visits based on the replacement usages of number of
injections, solvent pumped and lamp hours
Lower operating expenses through increased instrument
utilization
© 2008 Waters Corporation
36
W a t e r s UPLC® Pa r t icle s
Ove r vie w
Et hylene Bridged H ybrid ( BEH) Part icles
— Wide pH range (1- 12)
— Five chemistries
— Seamless HPLC ? UPLC® method migration – with same
selectivity as XBridgeTM HPLC columns
— 130Å and 300Å pore diameters
H igh St rengt h Silica ( HSS) Part icles
— ONLY UPLC® - certified 100% silica particle
— Three C18 chemistries
— Developed specifically for UPLC® applications
— Packed, t est ed and guarant eed com pat ibilit y wit h
pressures up to 15,000 psi (1000 bar)
© 2008 Waters Corporation
37
Th e Ch e m ist r ie s of UPLC ®
Te ch n ology
Launch
Date
Mar
2004
Mar
2005
Mar
2005
Mar
Dec
2005 2005
Sep
2006
Jun
2007
Dec
2007
© 2008 Waters Corporation
38
BEH Ch e m ist r ie s of UPLC ®
Te ch n ology
BEH C18
— Trifunctionally Bonded C 18
— First UPLC® column choice
— Superior peak shape &
efficiencies
BEH C8
— Trifunctionally Bonded C 8
— Wide pH range
BEH Shield RP18
— Monofunctionally bonded
— Embedded carbamate group
— Alternate selectivities
BEH Phenyl
— Trifunctionally Bonded C 6
( Hexyl) Phenyl
— Unique combination of
chemistry & particle
— Wide pH range
1.7 µ m
2.5, 3.5, 5, 10 µ m
BEH HILIC
— Unbonded, rugged BEH
particle
— HILIC for very polar bases
Ease of Migration from HPLC to UPLC ®
Simplified Purification and Isolation
© 2008 Waters Corporation
39
H SS Ch e m ist r ie s of UPLC ®
Te ch n ology
HSS T3
— T3: Polar compound retention
— Aqueous- compatible C18
chemistry
— Designed for maximum
retentivity
HSS C18
— High coverage, trifunctionally
bonded C18 chemistry
— Universal, high performance C18
chemistry
— Proprietary endcapping for
superior peak shape
— Silica particle performance
HSS C18 SB
— SB: Selectivity for Bases
— Non- endcapped: optimum
silanophilic selectivities
— Designed for method developers
© 2008 Waters Corporation
40