The new Agilent 1290 Infinity Quaternary LC How binary pump-like performance can enhance your lab efficiency Sebastian Krahe Produktspezialist HPLC Agilent Technologies Page 1
The 1290 Infinity Quaternary LC System Leveraging 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 Page 2
Innovation: 1290 Infinity Quaternary Pump Quaternary Pump with Binary Pump-like Performance 1290 Infinity Quaternary Pump Boost Performance Highest accuracy and precision for composition and flow, with exceptionally low delay volume Save Time Accelerates transfer of existing HPLC methods to UHPLC Reduce Costs Outstanding UHPLC performance at HPLC-like costs of ownership Page 3
1290 Infinity Quaternary Pump Various tools enabling the high performance Inlet-Weaver Switchable Jet Weaver Mixer 4-channel Degasser Multi-Purpose- Valve MCGV (Multi- Channel-Gradient- Valve) Pump Head In-line Filter Page 4
Multipurpose valve functions for highest comfort Standard Application for standard Delay Volume Page 5
Agilent 1290 Infinity Quaternary Pump Specifications & Benefits Power Range For 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 Composition Range 1-99 % Wide analytical range Delay Volume < 350 µl For fast quaternary gradients Page 6
Composition Accuracy and Precision Step Gradient Analysis with Tracer Different mixtures (H2O, MeOH, ACN) at different pressures and different flow rates Full application range! Page 7
Flow and Composition Accuracy and Precision Isocratic analysis of Glucocorticoids 1: Hydrocortisone 2: Beclomethasonedi-proprionate 3: Hydrocortisone acetate Column: Agilent ZORBAX Eclipse Plus C18, 4.6 x 100 mm, 5 µm (p/n 959996-902) Mobile phase: Water/ Acetonitrile = 30/70 Flow rate: 1 ml/min Stop time: 5 min Injection vol.: 3 µl (needle wash for 3 sec) Column Temp.: 30 C DAD: 254/10 nm, (ref 400/ 80 nm), 20 Hz, slit 4 nm Page 8
Flow and Composition Accuracy and Precision Isocratic analysis of Glucocorticoids mau 30 25 1 2 RT of compounds very sensitive regarding changes in mobile phase composition! 20 15 10 5 0 1 1.5 2 2.5 3 3.5 3 Water/ACN 29:71 Water/ACN 31:69 Water/ACN 30:70 min Page 9
Accuracy and Precision Isocratic analysis of Glucocorticoids Specification (< 0.07 %) mau 35 30 25 1: Hydrocortisone 2: Beclomethasonedi-proprionate 20 15 10 3: Hydrocortisone acetate 5 0 0 1 2 3 4 min Page 10
Accuracy and Precision Dynamically vs. pre-mixed solvents Isocratic composition by dynamic mixing: A/B = 70:30 Isocratic composition by premixed solvent: Which method is better? RSD RT(%) A/B = 70:30 Page 11
Accuracy and Precision Dynamically vs. pre-mixed solvents Retention time precision Single operator, single bottle Dynamic mixing For illustration only, data can be found in Agilent Application Note 5991-0098EN Page 12
Accuracy and Precision Dynamically vs. pre-mixed solvents Retention time precision Single operator, single bottle Dynamic mixing Single operator, multiple bottles For illustration only, data can be found in Agilent Application Note 5991-0098EN Page 13
Accuracy and Precision Dynamically vs. pre-mixed solvents Retention time precision Single operator, single bottle Dynamic mixing Single operator, multiple bottles Multiple operators, multiple bottles For illustration only, data can be found in Agilent Application Note 5991-0098EN Page 14
Accuracy and Precision Gradient Analysis of Clopidole Clopidole 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. 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 15min Flow rate: 1 ml/min Stop time: 15 min Injection vol.: 1 µl Column Temp.: 30 C DAD: 263/10 nm, (ref. 400/ 80 nm), 10 Hz Page 15
Accuracy and Precision Dynamically vs. pre-mixed solvents Gradient from 0 to 100 % with premixed solvent: H 2 O +0.1% TFA 0 100 % B ACN +0.1% TFA Gradient from 5 to 60 % B by dynamic mixing: H 2 O +0.1% TFA ACN +0.1% TFA 5 60 % B Which method is better? RSD RT(%) Page 16
Accuracy and Precision Dynamically vs. pre-mixed solvents Dynamic mixing 5 60 % B Premixed, 0 100 % B mau mau 200 150 150 100 100 50 50 0 0 6.0 6.4 6.8 7.2 7.6 min 6.0 6.4 6.8 7.2 7.6 min Page 17
Accuracy and Precision Dynamically vs. pre-mixed solvents Retention time precision 0.5 0.4 RSD RT (%) 0.3 0.2 0.1 0 Specification (< 0.15 %) 5 60 % B Dynamic mixing 0 100 % B Premixed Page 18
Agilent 1290 Infinity Quaternary Pump Specifications & Benefits Power Range For 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 Composition Range 1-99 % Wide analytical range Delay Volume < 350 µl For fast quaternary gradients Page 19
Ternary/Quaternary Gradients Blend Assist You need different concentrations of modifiers in your analysis, would like to have just one stocksolution and do online dilution to profite from the quaternary mixing capability of your pump? Here is a simple tool BlendAssist! Water Water 1% TFA Desired method conditions - example: 1. 5 to 95% gradient of ACN with 0.1% TFA in Water and 0.08% TFA in ACN 2. 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). ACN ACN 1% TFA With BlendAssist: just program your binary organic/aqueous gradient and define the dilution factor! Page 20
Ternary/Quaternary Gradients Blend Assist, different Phosphate buffer concentrations H 3 C CH 3 N H 3 C CH 3 N O NH 2 C H 3 N O OH C H 2 N Disopyramide Quinidine Column: Agilent ZORBAX Eclipse Plus C18, 4.6 x 100 mm, 1.8 μm Mobile phase: A = Water B = Water/Acetonirile 20:80 + 0.05 M NaH 2 PO 4, ph=4.6 Gradient: var. Flow rate: 1.5 ml/min Stop time: 8.5 min Injection vol.: 1 µl Column Temp.: 40 C DAD: 270/10 nm, (ref. 360/ 100 nm), 20 Hz Page 21
Ternary/Quaternary Gradients Blend Assist, different Phosphate buffer concentrations mau Quinidine 400 300 Procainamide Acetyl- Procainamide 0.05 M buffer 200 0.025 M buffer 100 0 Disopyramide 0.01 M buffer 0 1 2 3 4 5 6 7 8 min Page 22
Ternary/Quaternary Gradients Blend Assist - simple Tool for Online-Dilution of Modifiers and Gradient Set-Up BlendAssist - Solvent/modifier channel definition Page 23
1290 Infinity Quaternary Pump Conclusion 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! Page 24