new 1100 series dad sl and mwd sl 80hz data acquisition for ultra-fast lc
TRANSCRIPT
New 1100 Series DAD SL and MWD SL
80Hz Data Acquisition for Ultra-fast LC
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1100 Series DAD SL and MWD SL
Agilent 1100 Series Diode-array Detector SL Agilent 1100 Series Multi-wavelength Detector SL
The 1st Diode-array Detector designed for Ultra-fast LC
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1100 Series DAD SL and MWD SLOverview
Next Generation Electronics and Firmware provides
80Hz Data Acquisition of up to 8 Signals Up to 100% resolution gain in ultra-fast,
quantitative LC 80Hz Full Spectral Data Acquisition (DAD SL only)
for ultra-fast peak purity analysis and spectral conformation even for trace level compounds
Improved Diode-Array Front-end Electronics for minimized noise (typical < +/- 6μAU
ASTM) New Build-in Data Recovery Card
for a „data never lost insurance“ New RFID Tags for all Flow Cells and UV Lamp
for unambiguous data traceability LAN on Board
eliminates need for additional LAN interface Future proof design
Build-in web-server, USB, PCMCIA (WLAN, Bluetooth)
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1100 Series DAD SL and MWD SLOverview (cont’d)
New Electronic Temperature Control (ETC) For maximum practical sensitivity by
minimized baseline wander, especially under harsh and fluctuating ambient temperature and humidity conditions
New standard flow cell for maximum practical sensitivity by
minimized RI-sensitivity and dispersion
Builds upon 1100 DAD Optical Design preserves features like programmable slit
and dual lamp design for highest sensitivity from 190 to 950nm
Up to 20x Sensitivity from 400 – 950nm
+ 40% (+ 80%) Sensitivity by 8 and 16nm slit
Re-use spectral libraries of 1100 DAD „A“ and „B“
Confidence and robustness of a diode-array detector with more than 25,000 installations.
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What is Ultra-fast LC?
What is the Objective of Ultra-fast LC?
1.Significant gains in productivity, while maintaining or increasing data quality
• Ultra-fast LC provides up to 10x gains in analysis speed while preserving or increasing Resolution, Sensitivity, Linearity, Precision and Robustness.
• Thereby, Ultra-fast LC ensures compliance with strictest (regulatory) performance requirements.
2. Uncompromised compatibility with existing methods.
• Run conventional LC methods without compromising performance
• Comply with today’s and future requirements
3. Improved Data Security.
• Ultra-fast LC systems provide a new level of data security and traceability that prevents data losses and minimizes the risk of false data interpretation.
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• Conventional LC: Analysis Times = 5.0 – 120 min
• Fast LC: Analysis Times = 2.0 – 5.0 min
• Ultra-fast LC: Analysis Times = 0.2– 2.0 min
Gradient Time = 0.2 – 1.5 min
Cycle Times = 0.5 – 2.5 min
50% Peak Width = 0.1– 1.0 sec
PW = 0.3 sec
min0.1 0.2 0.3 0.4 0.50
How fast is Ultra-fast?
What is Ultra-fast LC?
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Which 1100 Configuration do I need for Ultra-fast LC?
1100 Series Ultra-fast LC System
• 1100 Series Binary Pump – for precise, high-pressure mixing gradient formation and low delay volume
• 1100 Series WPS – for precise, high-speed injection with lowest delay volume and carry over
• 1100 Series TCC – for precise, peltier-controlled high-temperature LC up to 80C
• 1100 Series DAD/MWD SL – for highest chromatographic resolution by 80Hz data rate
• Zorbax RRHT 1.8um Columns – for highest efficiency at high linear flows
What is Ultra-fast LC?
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min0.1 0.2 0.3 0.4 0.50
80Hz
PW=0.30sec
40Hz
PW=0.33sec
20Hz
PW=0.42sec
10Hz
PW=0.67sec
5HzPW=1.24sec
• Sample: Phenone Test Mix• Column: Zorbax SB-C18, 4.6x30,
1.8um• Gradient: 50-100% ACN in 0.3min• Flow cell: 5ul
80Hz versus 20Hz– 30% Peak Width+ 30% Resolution + 40% Peak Capacity+ 70% Apparent Column Efficiency
80Hz versus 10Hz– 55% Peak Width+ 90% Resolution+ 120% Peak Capacity+ 260% Apparent Column
Efficiency
What’s the Benefit of 80Hz Data Acquisition Rate?
Peak Width, Resolution and Peak Capacity in Ultra-Fast LC
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80Hz versus 20Hz Data Rate:– 30% Peak Width=> +40% Peak Capacity+30% Resolution => + 70% Apparent
Column Efficiency
80Hz versus 10Hz Data Rate:– 55% Peak Width => +120% Peak Capacity+90% Resolution => +260%
Apparent Column Efficiency
Data Rate
Peak Width
Resolution
Peak Capacity
80 Hz
0.300 2.25 61
40 Hz
0.329 2.05 56
20 Hz
0.416 1.71 44
10 Hz
0.666 1.17 28
5 Hz 1.236 0.67 16
0.2
0.4
0.6
0.8
1
1.2
1.4
0 20 40 60 80 100
Data Rate [Hz]
Peak
Wid
ths
/ se
c
0
10
20
30
40
50
60
70
Peak
Cap
acity
Peak Width [s]
Peak Capacity
0.2
0.4
0.6
0.8
1
1.2
1.4
0 20 40 60 80 100
Data Rate [Hz]
Peak
Wid
ths
/ se
c
0
0.5
1
1.5
2
2.5
Reso
lutio
n
Peak Width [s]
Resolution (4,5)
What’s the Benefit of 80Hz Data Acquisition Rate?
Peak Width, Resolution and Peak Capacity in Ultra-Fast LC
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Performance Requirements of Ultra-Fast LC
Ultra-fast LC using the 1100 DAD SL provides Resolution and Peak Capacity gains of up to 100%.
But – Can I still fulfill my (regulatory) performance requirements under ultra-fast LC conditions?
• Quantification of Side Products at 0.05% level ?
• RT Precision < 0.5% ?
• Area Precision < 1% ?
• Peak Purity Analysis at Trace Levels ?
• Spectral Conformation at Trace Levels ?
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Sensitivity and Linearity in Ultra-Fast LC Can I Simultaneously Quantity Main Compounds and Side
Products at 0.05% Level?
min0.5 1 1.5 2 2.5
mAU
0
500
1000
1500
2000
Main Compound = 2000mAU
Impurities = 1mAU
DMSO
Impurities
Is the detectors Dynamic Range large enough to accurately and precisely quantify Main
Compound and Impurities simultaneously?
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Sensitivity and Linearity in Ultra-Fast LC
Is the Noise Low Enough for my Quantitative Analysis?
ASTM Noise Specification:20 µAU Peak-to-Peak (+/- 10 µAU)
4nm Slit
8nm Slit
16nm Slit
80 Hz
42 31 23
40 Hz
30 22 16
20 Hz
21 16 11
10 Hz
15 11 8.0
2.5 Hz
7.4 5.4 4.0
• Temperature: 20C• DAD: 254nm,16nm, Ref 360,
80nm • PW: > 0.1min (2.5Hz, 2sec RT)
min5 10 15 20
mAU
-0.04-0.03-0.02-0.01
0
min5 10 15 20
mAU
-0.1-0.08-0.06-0.04-0.02
Conditions• Eluent: Water/ACN = 70/30• Flow rate: 1ml/min• Column: 4.6x30mm SB C18,
1.8um
min5 10 15 20
mAU
-0.06
-0.04
-0.02
0 4nm Slit widthNoise < +/– 3.7 µAU
8nm Slit widthNoise < +/– 2.7 µAU
16nm Slit widthNoise < +/– 2.0 µAU
Peak-to-Peak Noise on 13ul Flow Cell
– 27% – 26%
Note: 50µAU Noise gives S/N = 20 at 1mAU (0.05% level)
13
Sensitivity and Linearity in Ultra-Fast LC
Is the Linear Range Large Enough for my Quantitative Analysis?
Linearity (Caffeine Sample):
Deviation at 2.0AU:
• 2.0% (Vis Lamp off)
• 2.5% (Vis Lamp on)
5% Deviation:
• 2.5 AU (Vis Lamp off)
• 2.4 AU (Vis Lamp off)
Specification:
• 5% Deviation at 2.0 AU
80.0%
85.0%
90.0%
95.0%
100.0%
105.0%
0 500 1000 1500 2000 2500 3000
Absorbance / mAU
% D
evia
tion
from
Lin
ear V
alue
Vis Lamp off
Vis Lamp on
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•Gradient: 50–70% B in 0.85min
•Column: 4.6 x 50, 1.8um•Injection: 5ul of
550 µg/ml Nimodipin•Flow Rate: 4 ml/min•Flow cell: 13ul•Data Rate: 80Hz•Slit: 8nm
mAU
1800
1850
1900
1950
2000
0.6
28
0.6
27
0.6
27
0.6
27
0.6
27
0.6
27
0.6
27
0.6
28
0.6
27
0.6
27
min0.6250.6250.6260.6260.6270.6270.6280.6280.6291750
Overlay of 10 analyses at 245nm
• RT Precision: 0.067% RSD
• Area Precision: 0.13% RSD
Sensitivity and Linearity in Ultra-Fast LC
Reproducibility of Main Compounds at 2000mAU (100% Level)
min0.61 0.62 0.63 0.64 0.65 0.66 0.67
mAU
0
250
500
750
1000
1250
1500
1750
2000
Peak Width
0.70 sec
15
min0.2 0.25 0.3 0.35 0.4 0.45 0.5 0.55
mAU
0
1
2
3
4
5
6
Sensitivity and Linearity in Ultra-Fast LC
Reproducibility of Impurities and Side Products at Trace Level
Overlay of 10 analyses at 245 nm:
Nifedipin • A = 2.5mAU (0.1%
level)• RT Precision =
0.092% RSDNifedipin degradation
product • A = 0.5mAU (0.03%
level)• RT Precision =
0.123% RSD
•Column: 4.6 x 50, 1.8um
•Gradient: 50–70% B in 0.85 min
•Injection: 5ul•Flow Rate: 4 ml/min•Flow cell: 13ul•Data Rate: 80Hz•Slit: 8nm
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Nifedipin at trace levelsPeak Width = 0.63 sec
min0.3 0.31 0.32 0.33
mAU
0
0.5
1
1.5
22mAU = 0.1%
levelS/N = 50
1mAU = 0.05% level
S/N = 25
0.5mAU = 0.025% level
S/N = 12
Conditions:•Column: 4.6 x 50, 1.8um• Gradient: 50–70% B in 0.85 min• Injection: 5ul• Flow Rate: 4 ml/min• Flow cell: 13ul • Data Rate: 80Hz• Slit: 8nm
Result:Under ultra-fast LC conditions the DAD SL allows accurate quantitation of impurities and side products at levels smaller than 0.05% of the main compound(s).
Sensitivity and Linearity in Ultra-Fast LC
Can I quantity Impurities and Side Products at 0.05% Level?
Noise 40 µAU
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Apex Spectrum of Nifedipin at 0.1% (1.8 mAU) measured at 80 Hz:
nm300 400 500 600 700 800
mAU
0
0.5
1.0
1.5
nm300 400 500 600 700 800
mAU
0
400
800
1200
Reference Spectrum of Nifedipin at 100% level (1800 mAU) measured at 80 Hz:
Spectral Analysis in Ultra-Fast LCCan I do Peak Purity and Spectral Conformation at
Trace Levels?
Is the spectral quality obtained at trace level under ultra-fast LC conditions and 80Hz spectral sampling rate good enough for peak purity analysis and spectral conformation?
18
UV Spectrum of Nifedipin at higher concentration (ca. x180):
Spectral Analysis in Ultra-Fast LCCan I do 80Hz Peak Purity Analysis at Trace Levels?
min1.6 1.7 1.8 1.9 2
Norm.
6
8
10
12
14
DAD1 A, Sig=245,10 Ref=500,80 (N:\BACKUP_MSD\DATA_MRZ\MF_180305_STD_A_ALL\NIMIX_1_2000_1_G_05.D)
1.6
29
2.0
26
nm300 400 500 600 700 800 900
Fl
-0.25
0
0.25
0.5
0.75
1
1.25
1.5
1.75
mAU
Overlay of extracted Nifedipin spectra at trace level
Result: Nifedipin peak at 0.1% level (1.8mAU) measured with 80Hz spectral rate is pure
– no other compounds are co-eluting with Nifedipin
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nm300 400 500 600 700 800 900
Norm
0
0.25
0.5
0.75
1
1.25
1.5
1.75
2
nm300 400 500 600 700 800 900
Norm
0
0.25
0.5
0.75
1
1.25
1.5
1.75
2Overlay of 80Hz Reference and Apex Spectrum
•Nifedipin Ref: 1800mAU (100% level)
•Nifedipin Apex: 1.8mAU (0.1% level)
Spectral Analysis in Ultra-Fast LCCan I do 80Hz Spectral Conformation at Trace Levels?
Overlay of 80Hz Reference and Apex Spectrum
•Nimodipin Ref: 1800mAU (100% level)
•Nifedipin Apex: 1.8mAU (0.1% level)
Result:
Identification of Nifedipin at 0.1% trace level under fast LC conditions and 80Hz spectral sampling rate
Match Factor = 963
Match Factor = 929
20
min0 0.05 0.1 0.15 0.2 0.25 0.3 0.350
20
40
60
80
100
120
Performance• Average 50% Peakwidth =
0.34 sec• Resolution (4,5) = 1.5• Analysis Time = 24 sec • Cycle time = 50 sec• RT Precision = 0.7 – 0.22%
RSD• Area Precision = 1.5 – 0.3%
RSD
Conditions – An extreme Example• Sample: Phenone Test Mix• Column: 4.6 x 30mm, 3.5µm SB-C18• Gradient: 50-100% ACN in 0.3min• Flow rate: 5ml/min• Temperature: 40°C• Data Rate: 40Hz
Speed and Precision in Ultra-Fast LCPushing the Limits for highest Throughput
Peak RSD RT (%) RSD Area (%)
1 0.70 1.3
2 0.58 0.7
3 0.43 0.9
4 0.34 0.3
5 0.32 0.9
6 0.29 0.9
7 0.26 1.5
8 0.24 0.7
9 0.22 1.0
Overlay of 6 Runs
Application Areas• Screening
Experiments• HT LC/MS/UV• Early Formulation
Studies• Process Analytical
Techn.
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Speed and Precision in Ultra-Fast LCModerate Gradients for highest-quality Quantitative Data
Peak RSD RT (%) RSD Area (%)
1 0.47 0.472 0.44 0.193 0.31 0.304 0.27 0.275 0.25 0.486 0.23 0.427 0.20 0.338 0.17 0.359 0.15 0.27
min0.1 0.2 0.3 0.4 0.5 0.6 0.7
mAU
0
200
400
600
800
1000 Overlay of 6 Runs
• 2x slower gradient
• 2 – 5x better Precision
• Still very fast
Performance• Peakwidth = 0.38 sec FWHM• Resolution (4,5) = 1.6• Analysis Time = 40 sec • Cycle time = 70 sec• RT Precision = 0.1 – 0.5%
RSD• Area Precision = 0.2 – 0.5%
RSD
Conditions – Less extreme Condtions
• Sample: Phenone Test Mix• Column: 4.6 x 30mm, 3.5µm
SB-C18• Gradient: 50-100% ACN in
0.6min• Flow rate: 5ml/min• Temperature: 40°C• Data Rate: 40Hz
Application Areas• Formulation Studies• Analytical
Development• Process Control• QA/QC
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min0.2 0.4 0.6 0.8 1
injection 1
injection 4000
Column 2
Column 1injection 4000
injection 2000
injection 2000
injection 1
min0.2 0.4 0.6 0.8 1
Robustness in Ultra-Fast LCAre Methods and Instrumentation Robust Enough for
24x7 Operation?
Stability study on system configuration with automated column regeneration:
• Stable system and column performance for 8000 injection (4000 injections per column)
• System suitable for unattended and automated 24x7 operation and reliable over-weekend runs
Injection Number
0.4
0.5
0.6
0.7
0.8
0.4
0.5
0.6
0.7
0.8
0 500 1000150020002500300035004000
Column 1
Column 2
RT/min
W1/2 /sec
0 500 1000150020002500300035004000
RT/min
W1/2 /sec
P=300bar
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New Data Recovery Card*The first LC Detector with “Data Never Lost”
Insurance
*Patent Filed
Data Recovery Card* - DRC
• All signals, spectra and meta data are buffered on high-capacity, embedded 256MB Compact Flash Card compliant with 21 CFR Part 11.
• Prevents any data loss in case of communication breakdowns between instrument and PC.
• Automatic Run Recovery in case of temporary communication failures
• Manual Run Recovery in case of permanent communication failures” after software, PC, and/or instrument re-boot.
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New Data Recovery CardThe first LC Detector with “Data Never Lost”
Insurance
Automated Run Recovery in case of temporary communication failuresA) Time Elapsed = 0.3 min Run in Progress
B) Time Elapsed = 0.6 minCommunication Failure Occurs
C) Time Elapsed = 1.9 min • Communication is re-
established• Automatic data transfer from
DRC to PC• No user interaction necessary
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New Data Recovery CardThe first LC Detector with “Data Never Lost”
Insurance
Manual Run Recovery in case of permanent communication failures
• Run Recovery dialog pops-up automatically after system re-boot.
• Data are stored on the PC under a pre-configured location.
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The Next Level of Data Traceability Proprietary RFID Technology for Flow Cells and
UV Lamp
Radio Frequency Identification Tags
• RFID tags records all relevant data necessary to recall instrument conditions under which a run has been executed.
• Minimizes the risk of false data interpretation, because measurement conditions are documented.
• Meta data stored on RFID tags are saved with each raw data file for unambiguous answers to (auditor-) questions like
“Which type of flow cell was used to generate this chromatogram - what was the path length and volume?”
“Did the accumulated burn-time of the lamp exceed pre-defined limit?”
Flow Cell • Path length• Volume• Max pressure• Date last test passed• Product number• Serial number• Production date
UV Lamp • Accumulated on-time• Actual on-time• Number of ignitions• Date last test passed• Product number• Serial number• Production date
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The Next Level of Data Traceability Proprietary RFID Technology for Flow Cells and
UV Lamp
ChemStation Report
RFID-tag information documents run conditions.
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The Next Level of Data Traceability Proprietary RFID Technology for Flow Cells and
UV Lamp
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10 1010 RFID Tag Info for Diagnostics
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The next Level of Baseline StabilityNew Electronic Temperature Control – ETC
Electronic Temperature Control - ETC
Advantages of ETC
Compensation of changes of ambient conditions (temperature and humidity)
Reduced baseline wander for improved practical sensitivity and reproducibility under harsh environmental conditions
Optical Unit
Main Board
Optical Temp. Sensor
Ambient Temp. Sensor
Air Flow
Fan
Heater
1100 Series DAD SL / MWD SL
Front
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Conditions: Relative humidity = 60%RH = const;
Temp = 25°C +/-2°C; 4 x 1h CyclesNote: By keeping RH=const, the absolute humidity is strongly modulated due to temperature variations (worst case condition).
TempCntrl ON 254,4 No Ref 750,4 No Ref
TempCntrl OFF 254,4 No Ref 750,4 No Ref
2.6mAU / 4°C = 0.7mAU/°C
The next Level of Baseline StabilityNew Electronic Temperature Control – ETC
Ambient Rejection at
60% RH:
ETC off: ~
700µAU/°C
ETC on: < 30µAU/°C
Temp. C
RH %
AH g/kg
AH Dev. %
23 60 10.5 - 12
25 60 11.9 0
27 60 13.4 13
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TempCntrl ON 254,4 No Ref 750,4 No Ref
TempCntrl OFF 254,4 No Ref 750,4 No Ref
~0.5mAU/°C
~0.7mAU/°C
Conditions: Relative humidity = 95% RH = const;
Temp = 25°C +/-2°C; 4 x 1h CyclesNote: By keeping RH=const, the absolute humidity is strongly modulated due to temperature variations (worst case condition).
The next Level of Baseline StabilityNew Electronic Temperature Control – ETC
Ambient Rejection at
95% RH:
ETC off: ~
700µAU/°C
ETC on: < 30µAU/°C Temp. C
RH %
AH g/kg
AH Dev. %
23 95 16.9 - 12
25 95 19.1 0
27 95 21.5 13
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1100 DAD SL: 254,4 / No REF 1100 DAD SL: 254,4 /
360,100
1100 DAD B: 254,4 No REF
1100 DAD B: 254,4 / 360,100
~100µAU/°C
Comparison between 1100 DAD SL and 1100 DAD (“B”-model)
Conditions: Relative humidity = 60%RH = const;
Temp = 25°C +/-2°C; 4 x 1h CyclesNote: By keeping RH=const, the absolute humidity is strongly modulated due to temperature variations (worst case condition).
The next Level of Baseline StabilityNew Electronic Temperature Control – ETC
Ambient Rejection ETC
on:
DAD “B”: ~
100µAU/°C
DAD SL: < 30µAU/°C Temp. C
RH %
AH g/kg
AH Dev. %
23 60 10.5 - 12
25 60 11.9 0
27 60 13.4 13
33
- Insert- Body- Ring
New 13ul Standard Flow Cell Design*
Ceramic Ring for thermal de-coupling
*Patent Filed
“Drill” Design of Inlet and Outlet for faster flush-out
Advantages
Reduced RI-sensitivity
Improved peak dispersion
Minimized noise in in high-flow, high-temperature applications
34
New 13ul Standard Flow Cell Reduced RI Sensitivity
New Standard Flow Cell provides
3-4x lower RI-Sensitivity1100 DAD/MWD with old Standard Cell
1100 DAD SL/MWD SL with new Standard Cell
12 mAU
4 mAU
36 mAU
9 mAU
Demanding RI Test Gradient
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ColumnSample
TempFlow
WLEluent
Injection
SB-C18 1.0x50 Iso Std +H2NCSNH2
250.1 ml/min254,4; -/-80/20 H2O/ACN1 ul
New 13ul Standard Flow CellMinimized Dispersion for Maximum Resolution
and Sensitivity
Improved flush-out behavior of the new standard flow cell minimizes peak tailing thereby increasing peak heights and resolution.
new Standard Cellold Standard Cell
36
- Insert- Body- Ring
Flow Cell Portfolio of the 1100 DAD/MWD SL For Uncompromised Compatibility from Nano to Prep
Advantages – From Nano to Prep
Compatibility with Conventional LC, Ultra-fast LC, Capillary LC, Nano LC and Prep LC
Support of Analytical LC on Columns from 75 µm to 4.6mm ID
Support of Preparative LC on Columns from 4.6 to 50mm ID
RFID tags for Data Traceability and Diagnostics
Flow Cell Portfolio Standard: 13ul, 10mm path length,
120bar Semi-Micro: 5ul, 6mm path length,
120bar Micro: 1.7ul, 6mm path length,
400bar Semi-Nano: 500nl, 10mm path
length, 50bar Nano: 80nl, 6mm path length,
50bar Preparative: 3mm, 120bar Preparative: 0.3mm, 20bar Preparative: 0.06mm, 20bar
37
Which Flow Cell to use for Ultra-fast LC?
13µl Standard Flow Cell: For highest sensitivity and linearity High-demanding quantitative work,
e.g. analytical method development, QA/QC
4.6 – 3 mm ID Columns
1.7µl Micro Flow Cell: For highest selectivity Ultra-fast semi-quantitative work,
e.g. Screening Experiments, HT LC/MS/UV
2.1 – 1mm ID Columns
5µl Micro Flow Cell: Best compromise of sensitivity and
selectivity For good quantitative and qualitative
results, e.g. Screening, HT LC/MS/UV, Early Formulation Studies
4.6 – 1mm ID Columns
Flow Cell Volume/Pathle
ngth
Sensitivity &
Linearity
Resolution
13 µl / 10mm 100 % ~ 90 – 95% *
5 µl / 6mm ~ 75 – 85% *
~ 93 – 98% *
1.7 µl / 6mm ~ 65 – 75% *
100 %
* Depends on analytical conditions and column dimension
38
min5 10 15 20
mAU
-0.04
-0.03
-0.02
-0.01
0
Which Flow Cell to use for Ultra-fast LC? Noise Comparison
13ul Standard Flow CellNoise < +/- 2.1 uAU
5ul Semi-Micro Flow CellNoise < +/- 2.4 uAU
Conditions• Column: 4.6x30mm SB C18,
1.8um• Flow rate: 1ml/min• Mobile phase: Water• Temperature: 20C• DAD: 254nm, 16nm, Ref 360,
80nm• PW: > 0.1min (2.5Hz, 2sec
RT)• Slit = 16
min5 10 15 20
mAU
-0.04
-0.03
-0.02
-0.01
0
Specification:ASTM noise < +/- 10 uAU
Note:5ul cell shows similar noise as 13ul cell. However, the linear range is reduced due to the reduced path length of 6mm.
39
SensitivityLimit of Detection for Anthracene under Ultra-fast LC
Conditions
10pg Anthracene injected in 1ul 4nm slit
40Hz: LOD = 1.48pg
20Hz, LOD = 1.03pg
10Hz, LOD = 0.67pg
80Hz: LOD = 2.30pg
0 0.1 0.2 0.3 0.4
mAU
0
0.5
1
1.5
2
2.5
3
3.5
4
min
Performance: LOD/pg in Ultra-fast LCRetention Time = 12 sec, Peakwidth = 0.9
sec4nm Slit
8nm Slit
16nm Slit
80 Hz 2.30 1.68 1.24
40 Hz 1.48 1.08 0.80
20 Hz 1.03 0,75 0.56
10 Hz 0.67 0.49 0.36
5 Hz 0.48 0.35 0.26
– 27% – 26%
4nm Slit
8nm Slit
16nm Slit
2.5 Hz
1 0.73 0.53
Compare: Conventional LC on 1100 DAD/MWD „B“ Retention Time = 2 min, Peak width = 6 sec
40
1100 Series DAD SL and MWD SL – Builds upon 1100 DAD
Optical Design - Optimized for Best Sensitivity
Tungsten lamp Long-life
Deuterium Lamp
Holmium Oxide Filter
Flow Cell
Programmable slit Grating
1024 element
diode array
190 nm950 nm
Minimized Noise in
Visible WL-Range
Automated wavelength verification
Fast optimization of sensitivity and
resolution
Excellent wavelength resolution
More uptime > 2000h
1100 DAD – 25,000 Installations Worldwide
41
Tungsten Lamp Off:Noise: > 400 µAU
700nm
min0 1 2 3 4 5 6 7 8 9
mAU
-0.2
-0.1
0
0.1254nm
ACN/Water 20/800.2 ml/min1 nm
Eluent:Flow rate:Slit:
1100 Series DAD SL and MWD SL – Builds upon 1100 DAD
Dual Lamp Design for Highest Sensitivity from 190 to 950nm
Tungsten Lamp OnNoise: < 20 µAU
Tungsten Lamp Off:Noise: < 20 µAU
mAU
-0.20
0.20.40.6
min0 1 2 3 4 5 6 7 8 9
Tungsten Lamp OnNoise: < 20 µAU
Advantages Approx. constant Noise from
190 to 950nm
Up to 20x lower Limits of Detection for compounds absorbing in the visible range at λ > 400nm
Significantly higher confidence in qualitative, spectral analysis results
More accurate Peak Purity results, especially at trace levels.
More accurate Library Analysis and Spectral Confirmation results
42
1100 Series DAD SL and MWD SL – Builds upon 1100 DAD
Programmable Slit – Micromechanics
Motor Slit
1 Programmable Slit for 1, 2, 4, 8 and 16 nm
Advantages No need for manual slit changes
Fast optimization of sensitivity and resolution
Maximized Sensitivity by 16nm slit (+ 80% versus 4nm slit)
Maximized Spectral Resolution by 1nm slit (for optimized spectral analysis)
Documents slit width (GLP, data traceability)
43
nm230 240 250 260 270 280
ResolutionBenzene Spectrum at Trace Level0.7 mAU1 nm slit2 nm wavelength bunching
1 nm
2 nm
4 nm
8 nm
Absorbance
00.10.20.30.40.50.6
(mAU)
16 nm
1100 Series DAD SL and MWD SL – Builds upon 1100 DAD
Programmable Slit for fast Optimization of Sensitivity and Resolution
Noise level
44
Rapid Resolution HT 1100 Series Modification Kits
Converting an 1100 Binary System to Ultra-fast LC
3 Modification Kits
• 1100 – VWD: 4.6mm RRHT-1100 Series Ultra-fast LC Kit P/N 5188-5323
• 1100 – DAD: 4.6mm RRHT-1100 Series Ultra-fast LC Kit P/N 5188-5324
• 1100 – DAD/MS:2.1mm RRHT-1100 Series Ultra-fast LC Kit P/N 5188-5328
Content
• Filter
• Capillaries
• Fittings, Union
• UV Flow cell
• RRHT Columns
45
Rapid Resolution HT 1100 Series Modification Kits
Converting an 1100 Binary System to Ultra-fast LC
Fast LC Modifications for 1110 Binary LC System with DAD/MWD Detector and 4.6mm ID RRHT Columns
46
Summary1100 Series DAD SL and MWD SL
Faster resultswe
Higher Resolution
Higher Data Security
Higher Peak Capacity
80Hz Data Acquisition for up to 100% resolution gain in ultra-fast LC
High Precision, Linearity and Sensitivity to maintain data quality under ultra-fast LC conditions to comply with regulatory requirements to allow for spectral analysis at trace levels (DAD SL only)
High Instrument, Column and Method Stabilityenables robust 24x7 operation
Uncompromised Compatibility with Existing MethodsRun conventional methods without compromising data
quality Build-in Data Recovery Card
provides „data never lost insurance“ RFID Tags for Cells and UV Lamp
for unambiguous data traceability New Low Noise Electronics, New ETC, New Standard Flow
Cell for decreased short-term noise and increased practical
sensitivity
47
Appendix
48
Overview – Features and Specifications
Feature/Spec 1100 DAD 1100 DAD SL
1100 MWD 1100 MWD SL
Spectral DA Rate
10 Hz 80 Hz N/A N/A
Signal DA Rate
20 Hz 80 Hz 20 Hz 80 Hz
Data Recov. Card
No Yes No Yes
RFID tags No Yes No Yes
LAN on-board
No Yes No Yes
Noise 254nm +/- 10uAU +/- 10uAU +/- 10uAU +/- 10uAU
Noise 750nm +/- 10uAU +/- 10uAU +/- 10uAU +/- 10uAU
Noise dual WL
+/- 10uAU +/- 10uAU +/- 10uAU +/- 10uAU
Linearity > 2 AU > 2 AU > 2 AU > 2 AU
WL-Range 190-950nm 190-950nm 190-950nm 190-950nm
WL Accuracy +/- 1nm +/- 1nm +/- 1nm +/- 1nm
Dual –Lamp Yes Yes Yes Yes
Slit/nm 1,2,4,8,16 progr.
1,2,4,8,16 progr.
1,2,4,8,16 progr.
1,2,4,8,16 progr.
Note:
For uncompromised compatibility from nano-flow to preparative applications all detectors can be ordered with 5 different analytical flow cells (13ul, 5ul, 1.7ul, 500nl, 80nl) and 3 preparative flow cells (3mm, 0.3mm and 0.06mm)
49
Future proof design
• State-of-the-Art Motorola PowerPC Processor
• LAN on board
• USB on board
• PCMCIA• Firewire• WLAN• Bluetooth
• Integrated Web-server
Module Independent Electronic Core
New Electronics platform
50
Peak Width – Response Time – Data Rate and Sensitivity
Don‘t use for > 0.15 sec peak width.
> 0.15 sec
> 0.3 sec
> 0.6 sec
> 1.2 sec
> 3 sec
> 6 sec
> 12 sec
> 24 sec
> 51 secPeak Width = Peak Width at 50% Peak Height
Recommended settings in ultra-fast LC with 50% peak width between 0.15 and 0.6 sec For 50% peak width between 0.6 and 1.2 secNotes: •Noise level changes ~ proportional to the
square root of the change in data rate.•For optimum selectivity and sensitivity
the Peak Width should not be chosen smaller than necessary.
•For 50% peak width between 0.3 and 0.6 seconds Peak Width of > 0.005 min is recommended, which correspondes to 40Hz data rate.
•For peaks narrower than 0.3sec at half height, Peak Width of > 0.0025min (80Hz data rate) should be used.
•For highest sensitivity in ultra-fast LC the slit can be increased to 8 or 16nm.
51
Linearity - OQ/PV Test on Caffeine Standards
min3 3.2 3.4 3.6 3.8
mAU
0
200
400
600
800
1000
Amount[ng/ul]0 200
Area
0
1000
2000
3000
4000
12
3
4
5
caffeine, DAD1 A
Correlation: 1.00000
Rel. Res%(1): 16.123
Area = 8.39574204*Amt +5.3180278
Correlation:
1.00000
52
Agilent 1100 Series HT LC/MS SystemA Scalable and Flexible Solution for Ultra-fast
Analysis
1.
5.
2.
3.
4.
6.
1. HT LC System + 1.8um RRHT Columns
2. Capacity Extension
3. Valves
4. Mass-selective Detector
5. Integrated Controller
6. Services and Compliance Products
53
-0.0005
0.0000
0.0005
0.0010
0.0015
0.0020
0.0025
0.0030
0.0035
0.0040
0.0045
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6
Van Deemter Curves1.8um Rapid Resolution HT Columns on the
1100 HT System
ZORBAX Eclipse XDB-C18 4.6 x 50mm (30mm) 85:15 ACN:Water 1.0L Octanophenone 0.05 – 5.0 mL/min 20°C
5.0 m
3.5 m
1.8 m
260,740 N/m @ 2mL/min
5.0 mL/min
Efficiency gain of 1.8μm versus 5μm columns: 3.2x @ 2ml/min4.4x @ 5ml/min
HETP
(cm
/pla
te)
Interstitial linear velocity (ue- cm/sec)
Higher speed – Higher resolution – Higher sensitivity Particle H_min
5μm 9.3μm
3.5μm 6.0μm
1.8μm 3.8μm
Note: Efficiency of 1.8μm columns is virtually flow-rate independent.
Up to 2.1x Resolution Up to 4.4x Speed
54
Ultra-fast Gradient Analysisof 9 Phenones
Performance Summary
9 Phenones baseline separated in 29 sec39 sec0.8 min (with column regeneration and run time = 0.7min)1.0 min (without column regeneration and run time = 0.7min)> 2.65 for all peaks0.50 sec (average)79 (0.65min gradient)< 0.2% RSD without column regeneration< 1.0% RSD with column regeneration< 0.003% (limit of detection)7.2 days or 8000 injections of continuous operation with stable
performance
Analysis speed
Analysis timeCycle time
Resolution 4 peakwidthPeak capacityRT precision
Carry over Robustness
min0 0.2 0.4 0.6 0.8 1
mAU
0
100
200
300
400
Possible cycle time with CR = 0.8 min
Analysis time = 0.65 min = 39 sec
Possible cycle time w/o CR = 1.0 min
60°C4.35ml/min390bar