Infinity Quaternary LC can enhance your lab efficiency · Page 1 The new Agilent 1290 Infinity...
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Page 1 The new Agilent 1290 Infinity Quaternary LC How binary pump-like performance can enhance your lab efficiency Sebastian Krahe Produktspezialist HPLC Agilent Technologies
Transcript of Infinity Quaternary LC can enhance your lab efficiency · Page 1 The new Agilent 1290 Infinity...
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The new Agilent 1290 Infinity Quaternary LC
How binary pump-like performance can enhance your lab efficiency
Sebastian KraheProduktspezialist HPLCAgilent Technologies
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The 1290 Infinity Quaternary LC SystemLeveraging the Performance of the 1290 Infinity Binary LC System
1290 Infinity DAD with 60mm cell (& HDR optional)
1290 Infinity TCC with broadest valve selectivity
1290 Infinity Autosampler (with Flexible Cube for lowest carry-over)
1290 Infinity Quaternary Pump
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Innovation: 1290 Infinity Quaternary PumpQuaternary Pump with Binary Pump-like Performance
1290 Infinity Quaternary Pump
• Boost PerformanceHighest accuracy and precision for composition and flow, with exceptionally low delay volume
• Save TimeAccelerates transfer of existing HPLC methods to UHPLC
• Reduce CostsOutstanding UHPLC performance at HPLC-like costs of ownership
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1290 Infinity Quaternary PumpVarious tools enabling the high performance
4-channel Degasser
MCGV (Multi-Channel-Gradient-Valve) Pump Head
Multi-Purpose-Valve
Switchable Jet Weaver Mixer
In-line Filter
Inlet-Weaver
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Multipurpose valve functions for highest comfortStandard Application for standard Delay Volume
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Power RangeFor any kind of analysis
Composition Accuracy and Precision< 0.15 % RSD or 0.02 min SD± 0.4 % (1-99 % Composition B)
High RT precision in gradient runs
Flow Accuracy and Precision< 0.07 % RSD or 0.01 min SD± 1.0 % or 10 µL
High RT precision in isocratic runs
Agilent 1290 Infinity Quaternary PumpSpecifications & Benefits
Composition Range1-99 %
Wide analytical range
Delay Volume< 350 µL
For fast quaternary gradients
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Different mixtures (H2O, MeOH, ACN) at different pressures and different flow rates
Full application range!
Composition Accuracy and PrecisionStep Gradient Analysis with Tracer
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Flow and Composition Accuracy and PrecisionIsocratic analysis of Glucocorticoids
3: Hydrocortisoneacetate
1: Hydrocortisone 2: Beclomethasone-di-proprionate
Column: Agilent ZORBAX Eclipse Plus C18, 4.6 x 100 mm, 5 µm (p/n 959996-902)
Mobile phase: Water/ Acetonitrile = 30/70Flow rate: 1 mL/minStop time: 5 minInjection vol.: 3 µL (needle wash for 3 sec)Column Temp.: 30 °CDAD: 254/10 nm, (ref 400/ 80 nm), 20 Hz, slit 4 nm
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Flow and Composition Accuracy and PrecisionIsocratic analysis of Glucocorticoids
min1 1.5 2 2.5 3 3.5
mAU
0
5
10
15
20
25
30
Water/ACN 29:71
1
2
3
Water/ACN 31:69Water/ACN 30:70
RT of compounds very sensitive regarding changes in
mobile phase composition!
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Accuracy and PrecisionIsocratic analysis of Glucocorticoids
min0 1 2 3 4
mAU
0
5
10
15
20
25
30
35
3: Hydrocortisoneacetate
1: Hydrocortisone
2: Beclomethasone-di-proprionate
Specification (< 0.07 %)
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
A/B = 70:30
Isocratic composition by dynamic mixing:
Isocratic composition by premixed solvent:
A/B = 70:30
Which method is better?RSD RT(%)
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
For illustration only, data can be found in Agilent Application Note 5991-0098EN
Single operator,single bottle
Dynamic mixing
Retention time precision
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
For illustration only, data can be found in Agilent Application Note 5991-0098EN
Single operator,single bottle
Single operator,multiple bottles
Dynamic mixing
Retention time precision
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
For illustration only, data can be found in Agilent Application Note 5991-0098EN
Single operator,single bottle
Single operator,multiple bottles
Multiple operators,multiple bottles
Dynamic mixing
Retention time precision
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Accuracy and PrecisionGradient Analysis of Clopidole
Clopidole
Column: Agilent ZORBAX SB-C18, 4.6 x 250 mm, 3.5 µm (p/n 884950-567)Mobile phase: A = Water/ACN (95/5) + 0.1 % TFA
B = Water/Acetonitrile(40/60) + 0.1 % TFA
Gradient: 0 – 100 % B in 15minFlow rate: 1 mL/minStop time: 15 minInjection vol.: 1 µLColumn Temp.: 30 °CDAD: 263/10 nm, (ref. 400/ 80 nm), 10 Hz
Clopidole is used as prophylactics and/or therapeutics in the prevention or treatment of coccidiosis in the poultry and rabbit industries. It is added to finished feed in a concentration range of 100 to 125mg/kg.
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
Which method is better?RSD RT(%)
5 60 % B
Gradient from 5 to 60 % B by dynamic mixing:
H2O+0.1% TFA
ACN+0.1% TFA
Gradient from 0 to 100 % with premixed solvent:
0 100 % BH2O
+0.1% TFA
ACN+0.1% TFA
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
min6.0 6.4 6.8 7.2 7.6
mAU
0
50
100
150
200
min6.0 6.4 6.8 7.2 7.6
mAU
0
50
100
150
Dynamic mixing 5 60 % B Premixed, 0 100 % B
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Accuracy and PrecisionDynamically vs. pre-mixed solvents
0
0.1
0.2
0.3
0.4
0.5
Dynamic mixing Premixed
RSD
RT
(%)
5 60 % B
Specification (< 0.15 %) 0 100 % B
Retention time precision
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Power RangeFor any kind of analysis
Composition Accuracy and Precision< 0.15 % RSD or 0.02 min SD± 0.4 % (1-99 % Composition B)
High RT precision in gradient runs
Flow Accuracy and Precision< 0.07 % RSD or 0.01 min SD± 1.0 % or 10 µL
High RT precision in isocratic runs
Agilent 1290 Infinity Quaternary PumpSpecifications & Benefits
Composition Range1-99 %
Wide analytical range
Delay Volume< 350 µL
For fast quaternary gradients
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Ternary/Quaternary GradientsBlend Assist
You need different concentrations of modifiers in your analysis, would like to have just one stock-solution and do online dilution to profite from the quaternary mixing capability of your pump? Here is a simple tool – BlendAssist!
Water Water 1% TFA
ACNACN
1% TFA
Desired method conditions - example: 1. 5 to 95% gradient of ACN with 0.1% TFA in
Water and 0.08% TFA in ACN2. 20 – 80% gradient of ACN with 0.5% TFA in
Water and 0.4% TFA in ACN
Without BlendAssist you need to either pre-mix the required solvents or by using stock-solutions of TFA in Water and ACN to program complex gradients (%A, B, C, D).
With BlendAssist: just program your binary organic/aqueous gradient and define the dilution factor!
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Ternary/Quaternary GradientsBlend Assist, different Phosphate buffer concentrations
Column: Agilent ZORBAX Eclipse Plus C18, 4.6 x 100 mm, 1.8 μmMobile phase: A = Water
B = Water/Acetonirile 20:80 + 0.05 M NaH2PO4, pH=4.6Gradient: var.Flow rate: 1.5 mL/minStop time: 8.5 minInjection vol.: 1 µLColumn Temp.: 40 °CDAD: 270/10 nm, (ref. 360/ 100 nm), 20 Hz
Quinidine
N
OH
N
OCH3 CH2
N
NH2
ON
CH3
CH3
CH3
CH3
Disopyramide
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Ternary/Quaternary GradientsBlend Assist, different Phosphate buffer concentrations
min0 1 2 3 4 5 6 7 8
mAU
0
100
200
300
400
0.05 M buffer
0.025 M buffer
0.01 M buffer
Procainamide Acetyl-Procainamide
Disopyramide
Quinidine
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Ternary/Quaternary GradientsBlend Assist - simple Tool for Online-Dilution of Modifiers and Gradient Set-Up
BlendAssist - Solvent/modifier channel definition
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1290 Infinity Quaternary PumpConclusion
Full 1290 Infinity LC Power range
Highest composition precision and accuracy, solvent independent and across pressure and flow rate range
Full application range with binary-like pump performance!
Blend Assist for Buffer/Additive Blending
ISET-ready!
Full flexibility!
