The Right Solution The Agilent 1200 Infinity Series

The Right Solution: The Agilent 1200 Infinity Series for LC Analysis Laura Phillips Application Scientist Agilent Technologies June 27 th, 2013 Page

The Agilent 1200 Infinity Series Where Do We Come From? 1100 Series 1200 Series (1995 – 2006) (2006 – 2010) 1100 Bin/Quat HPLC Bin (HPM) Quat (LPM) 400 bar 5 m. L/min 1200 RRLC UHPLC Bin (HPM) 600 bar 5 m. L/min Price: 1200 Bin+20% 1200 Bin/Quat HPLC Bin (HPM) Quat (LPM) 400 bar 5 m. L/min 1200 Infinity Series 1290 Infinity (2009) UHPLC Bin (HPM) Quat (LPM - 2012) 1200 bar 5 m. L/min 1260 Infinity Bin/Quat UHPLC Bin (HPM) – 5 m. L/min Quat (LPM) – 10 m. L/min 600 bar Price ~ 1200 Bin/Quat 1220 Infinity UHPLC Bin (LPM) 600 bar, 5 m. L/min Price < 1200 Quat Page 2 June 27, 2013

The Agilent 1200 Infinity Series Raising the Standard in HPLC – UHPLC for EVERY Lab 1200 Infinity Series 1260 Infinity 1220 Infinity Low-flow Cap. LC Isocratic, manual Analytical flow Binary 1290 Infinity Prep-flow Binary Nano-LC Gradient, manual new Quaternary Chip-LC Gradient, automated Isocratic new Gradient-DAD Binary-SFC Quaternary Bio-Inert Page 3 June 27, 2013

The Agilent 1200 Infinity Series Raising the Standard in HPLC – UHPLC for EVERY Lab Method Developement Solution Multi-method Solution High-Dynamic Range DAD 2 D-LC Solution Online-SPE Valve solutions Bioinert solutions SFC solutions GPC solutions Page 4 June 27, 2013

1220 Infinity LC A solution to all your needs • Dual gradient pump with integrated degasser, 600 bar, up to 10 m. L/ min, for flexible and reliable analysis • Vial sampler with injector programming for higher automation • 80°C oven for one 25 cm column for better results • Variable wavelength or diode array detector with 80 Hz sampling rate for high sensitivity • Easy: Small footprint, low height. One cable. Affordable with Chem. Station/ EZChrom VL software Expandable: Any other module can be added • • Page 5 June 27, 2013

600 bar for HPLC and UHPLC Same as with the 1260 Infinity LCs, existing conventional HPLC methods from 1100/1200 Series LC can be run on the 1220 Infinity LC and giving the same results as well as UHPLC methods on 3. 0 mm and 4. 6 mm. ID columns can be run. Both with excellent performance! Saving half of the time by transferring a HPLC method to UHPLC, without loss of resolution. Excellent retention and area RSDs even under UHPLC conditions. Page 6 June 27, 2013

Expandability External tray for walk-up: Together with Agilent‘s Easy. Access software the 1220 Infinity LC with external tray is a perfect solution for walk-up environments! Additional detectors: Almost all 1260/1290 Infinity detectors and SQ-MS can be added! Page 7 June 27, 2013

Innovation: 1220 Infinity Mobile LC Solution Affordable, Flexible, Compatible – Everywhere! Instrument Control Framework • Boost Performance Complete UHPLC system for higher resolution and shorter run times • Save Time Bring your laboratory to the samples • Reduce Costs Most affordable system giving you access to UHPLC technology Page 8 June 27, 2013

1260 Infinity LC 1260/1290 Valve Solutions 1260 Infinity Bio-Inert LC – Buffer Advisor 1260 Infinity SFC 1260 Infinity GPC 1260/1290 ELSD Page 9 June 27, 2013

1200 Infinity Series valve solutions Page 10

The largest and most flexible valve portfolio Workflow- and Application-Automation Flexibility Types 600 bar, 1200 bar Std, Bio-inert, µ-flow, prep NEW Quick-Change Valve heads for universal Valve Drive 2 pos/4 port duo 2 pos/ 6 port 2 pos/10 port 4 pos/10 port 6 pos/14 port 8 pos/ 9 port 12 pos/13 port 6 pos/ 7 port 2 pos/ 8 port 2 pos/12 port Custom Valves 1290 Infinity TCC 1290 Infinity Flexible Cube Applications • Multi-method/Method Dev • Column switching (2, 4, 6, 8) • Solvent selection (13 – 26) • Matrix-stripping • Sample Enrichment • Online-SPE • Detector selection • Heart Cutting 2 DLC • Comprehensive 2 DLC • Alternating Column Regen. • Process Control/ Sample Sampling/ Fractionation • Column Connection 1290 Infinity Valve Drive Page

Automated Column Regeneration 2 -position / 10 -port valve head • 1 x 1290 Infinity Thermostatted column compartment (or 1290 Infinity Valve Drive) • 1 x 2 -pos/10 -port valve head • While the analysis is running on column 1, column 2 is regenerated using the flow from a second pump • After the analysis on column 1, the flow switches to column 2 for analysis while column 1 is regenerated. Page 12

Sample Enrichment 2 -position / 6 -port valve head • 1 x 1290 Infinity Thermostatted column compartment (or 1290 Infinity Valve Drive) • 1 x 2 -pos/6 -port valve head • The analyte is retained and concentrated on an enrichment column while the sample matrix flows to waste. • For analysis, the concentrated sample is flushed out of the enrichment column to the analytical column. Page 13

Sample Cleanup 2 -position / 6 -port valve head • 1 x 1290 Infinity Thermostatted column compartment (or 1290 Infinity Valve Drive) • 1 x 2 -pos/6 -port valve head • Matrix components are retained on a pre-column while the analytes pass through to the analytical column for separation. • After sample cleanup or matrix stripping, the precolumn is flushed in reversed flow direction by the second pump. Page 14

Column Selection for Method Development 6 -position / 14 -port valve head • 1 x 1290 Infinity Thermostatted column compartment • 1 x 6 -pos/14 -port valve head • Up to 6 columns (4 columns recommended because of available space) Page 15

Solvent Selection for Method Development 12 -position selection valve head Typically used with 1290 Infinity Valve Drive. For Method Development systems: 1290 Infinity Valve Drive and new 12 -position selection valve head is fully supported as pump-valve cluster and by Agilent Chem. Station Method Scouting Wizard Page 16

1260 Infinity Bio-Inert LC Page 17 June 27, 2013

Innovation: 1260 Infinity Bio-inert LC Truly Bio-inert UHPLC System 1260 Infinity Bio-inert LC • Boost Performance Highly versatile (U)HPLC system for small- and largemolecule analysis • Save Time Stainless steel system Fast switching between bioand non-bio applications, including cleaning • Reduce Costs Only one system to maintain for bio- and non-bio analysis 1260 Infinity Bio-inert LC Page 18 June 27, 2013

New Capillary Design Ensures Bio-inertness Capillaries Metal clad PEEK capillary Newest fitting design: Hybrid technique New capillary technolgy enables 600 bar AND is completely metal free !! Page 19 June 27, 2013

Buffer Advisor Software Facilitates Workflows in Ion Exchange Chromatography Manual preparation: 1. Tedious 2. Error-prone 3. Irreproducible (operator) Buffer Advisor helps to: 1. Define stock solutions for salt and p. H gradients 2. Check if the buffer system is suitable for your separation task 3. Create a corrected time table for a salt- or p. H gradient. Part of Bio-inert pump, G 5611 A Page 20 June 27, 2013

Buffer Advisor User Interface Area 1: Select Buffer and Gradient Type Area 2: Enter Gradient Time Table Area 3: Gradient Display Section: Process Gradient and Review Optimized Gradient with Compensation Steps Area 4: Review Composition of Stock Solutions and Adjust Page 21 June 27, 2013

1260 Infinity SFC Page 22 June 27, 2013

Innovation: Hybrid SFC/UHPLC System Double your Discovery Hybrid SFC/UHPLC System • Boost Performance SFC with LC-like sensitivity, unique 600 bar x 5 m. L/min power range • Save Time Efficient instrument utilization and orthogonality through unique switching capability between LC and SFC • Reduce Costs Lower solvent expenses, less waste, and only one system to maintain. Upgrade from LC to SFC Page 23 June 27, 2013

Agilent 1260 Infinity Analytical SFC/MS System 1260 Infinity SFC and the G 6100 SQ Mass Spectrometer Analysis of leachables in food packaging material by SFC/MS Page 24 June 27, 2013

1260 Infinity GPC Page 25 June 27, 2013

High Performance GPC/SEC for Routine Work High Performance GPC/SEC Solution • Boost Performance High performance pump & oven for reproducible molecular weights • Save Time RID warm up less than 1 hour, “I have always been impressed by the quality, robustness, reliability and easeof-use of their products, which have served me well. ” - John P. O’Donnell, Research Scientist, FUJIFILM Imaging Colorants Ltd. , Manchester, UK complete system based on industry-leading Agilent LC systems for maximum uptime • Reduce Costs RID with built-in solvent recycle, minimum training effort. Page 26 June 27, 2013

1260 Infinity GPC/SEC Multi Detector Suite Summary Key Benefits Comprehensive Allows complex multi detector GPC experiments to be performed routinely. Versatile The MDS can be controlled by software or the keypad for maximum ease of use, and can be used with any existing modular or integrated HPLC system. Convenient Data from an existing detector can be collected and processed through the MDS. High Quality Results The detector cells are heated for optimum baseline stability and minimum instrument downtime. Adaptable Can be expanded at any time to include additional detectors. Safe Incorporates multiple solvent leak detectors to prevent spillage of potentially hazardous solvents. Page 27 June 27, 2013

Agilent 1260 Infinity ELSD & 1290 Infinity ELSD Evaporative Light Scattering Detection (ELSD) Page

Introduction: ELSD General Benefits • Universal detection • More uniform response • Gradient Compatible • Complimentary to LC-MS • Eliminates derivatization steps in analytical methods Page 29 June 27, 2013

Agilent ELSD Portfolio G 4260 B G 4261 B 1290 Infinity ELSD (Cooling deleted) 1260 Infinity ELSD Light Source Blue LED (480 nm) Temperature range Nebulizer: 25 -90°C Evaporator: ambient – 120°C • Features • Hydrocortisone limit of detection 30 ng RS 232 Communication only Blue Laser (405 nm) Nebulizer: 25 -90°C Evaporator: ambient – 120°C • • • 1290 Infinity ELSD (Standard, cooled) Nebulizer: 25 -90°C Evaporator: 10°C – 80°C Hydrocortisone limit of detection 4 ng 5 -9 x improvement in sensitivity over current model Unique sub-ambient operation (greater response for volatile compounds) RS 232 and LAN Communication Voltage selection (110/240 V) no longer required Variable gas flow for uniformity of response Fast data collection (up to 80 Hz) Controlled via Chem. Station / EZChrom Front Panel control of all parameters (Temperature, gas flow, data rate etc) Page 30 June 27, 2013

Increased sensitivity, reduced baseline noise The increased laser intensity coupled with a high gain photomultiplier and digital signal processing increases signal and reduces noise. Signal/noise calculated using Open. Lab Chemstation v 1. 04 Injection: 10µL (Hydrocortisone 50 ng-on column) Column: Zorbax Eclipse XDB-C 18 (5µm, 4. 6 x 150 mm) Mobile Phase: Water/ACN (80/20) Flow Rate: 1. 0 m/min ELSD: Neb 40°C, Evap 60°C, Gas 1. 05 SLM G 4261 B S/N 158 G 4260 B S/N = 5 Page 31 June 27, 2013

Triglycerides in Natural Oils Sample: 2 mg/ml Starflower Oil Column: C 18 5µm, 250 x 4. 6 mm Eluent A: ACN Eluent B: DCM Gradient: 30 -50% B in 40 mins; 50 -90% in 2 mins, hold for 3 mins Flow Rate: 1. 0 ml/min Inj Vol: 20µl Detector: neb=40°C, evap=70°C, gas=1. 4 SLM Page 32 June 27, 2013

Environmental Application: Characterizing Biodiesel Column: BDS Hypersil Cyano 3µm, 250 x 4. 6 mm Eluent A: 0. 4% Acetic acid in Hexane Eluent B: 0. 4% Acetic acid in Methyl tert-Butyl Ether (MTBE) Gradient: 100% A, hold 5 mins; 0 -80% B in 10 mins, hold 2 mins; 80 -0%B in 3 mins Flow Rate: 1. 0 ml/min Inj Vol: 20µl Detector: neb=40°C, evap=80°C, gas=0. 8 SLM Page 33 June 27, 2013

1290 Infinity LC 1290 Infinity Quaternary Pump 1290 2 D LC 1290 Infinity with ISET Page 34 June 27, 2013

1290 Infinity Quaternary LC Page 35 June 27, 2013

Innovation: 1290 Infinity Quaternary Pump with Binary Pump-like Performance 1290 Infinity Quaternary Pump • Boost Performance Highest, binary-like accuracy and precision for composition and flow, with exceptionally low delay volume • Save Time Accelerates method transfer from R+D to QA and from HPLC to UHPLC • Reduce Costs One pump for any applications: Most universal – at HPLC-like costs of ownership Page 36 June 27, 2013

Agilent 1290 Infinity Quaternary Pump Specifications & Benefits Power Range Ø 1200 bar up to 2 m. L/min Ø 800 bar at 5 m. L/min Ø For any kind of analysis Flow Accuracy and Precision < 0. 07 % RSD or 0. 01 min SD ± 1. 0 % or 10 µL Ø High RT precision in isocratic runs Composition Accuracy and Precision < 0. 15 % RSD or 0. 02 min SD ± 0. 4 % (1 -99 % Composition B) Ø High RT precision in gradient runs Composition Range 1 -99 % Ø Wide analytical range Delay Volume < 350 µL Ø For fast quaternary gradients Page 37

1290 Infinity 2 D-LC Solution Page 38 June 27, 2013

Innovation: 1290 Infinity 2 D-LC Solution Square your Separation 1290 Infinity 2 D-LC Solution • Boost Performance New levels of separation power, detection sensitivity and precision in 2 D-LC • Save Time A single ultra high peak capacity run instead of many conventional runs – with 1 min method setup! • Reduce Costs Special features to reduce solvent and valve cost, upgrade paths for existing LC-systems Page 39 June 27, 2013

Data Analysis For heart-cutting 2 D-LC by Agilent‘s Open. LAB CDS Chem. Station ed. For comprehensive 2 D-LC Agilent recommends: GC Image LCx. LC Edition Software from GC Image LLC. Key features offered: • Direct Agilent datafile import • Peak identification, integration, annotation • Comparative Analysis and Visualization • Compound-libraries • MS-data handling Page 40 June 27, 2013

Hardware Module-flexibility 2. Dimension One or two 1290 Infinity TCC 1. Dimension 1290 Infinity Binary Pump 1260 Infinity Capillary Pump 1260 Infinity Binary or Quaternary Pump 1290 Infinity Binary Pump 1290 Infinity Autosampler or 1260 Hi. P Autosampler 1260 Infinity Autosampler Optional 1260/1290 Infinity Detector For 1 st dimension chromatogram and peaktriggering 1260/1290 Infinity detector To monitor waste-line Almost any Agilent pump or autosampler in the 1 st dimension! Almost any detectors are supported! A 1290 Infinity Binary Pump for the 2 nd dimension is required. Page 41 June 27, 2013

Applications examples - components of beer samples Analysis of two different commercially available beer samples with different separation modes 1 st dimension - Reverse Phase (C 18) 2 nd dimension - Reverse Phase (Phenyl)　 Beer B Separation by RP (Phenyl) Beer A Separation by RP (C 18) Page 42 June 27, 2013

1290 Infinity Binary LC w. ISET Page 43

Innovation: ISET Intelligent System Emulation Technology Intelligent System Emulation Techn. • Boost Performance The same retention times and peak resolution as the target system • Save Time No method redevelopment or system modification required • Reduce Costs Minimized investment and service costs, with one system for all methods Page 44 June 27, 2013

What is Method Transfer? Different Aspects From particle size to particle size (HPLC to UHPLC) ð often associated with a change of instruments due to constraints in max. pressure, extracolumn band broadening etc. . Some fine-tuning of method might be required due to frictional heating effects ð available tools : Agilent Method translator, Third Party Method development SW (ACD Labs, Chrom. Sword etc. ) From one eluent type or phase chemistry to an other ð Method development ð available tools: Method development SW (Chromsword, ACD Labs, etc. ) From column dimension to column dimension (e. g. 4. 6 to 2. 1 mm i. d. , 50 mm to 100 mm) ð recalculate flow rates, recalculate gradient times, adjust connection capillaries and flow cells, due to delay volume results may vary, method might need revalidation ð available tools: Agilent Method translator From Instrument to Instrument ð with method/instrument change: Isocratic Hold / Pre-injection, Vol. Modification ð available tools w/o method or instrument change : none until ISET Page 45 June 27, 2013

Publication number 5990 -9546 EN Publication number 5990 -9703 EN Publication number 5990 -9545 EN Publication number 5990 -9703 EN Intelligent System Emulation Technology Agilent Technical and Application Notes Page 46

Instrument Control Framework Page 47 June 27, 2013

Innovation: Instrument Control Framework Integrate your Agilent LC with other CDS Instrument Control Framework • Boost Performance Full functionality of all Agilent LC Systems in non-Agilent data systems • Save Time Fast plug-and-play updates to latest Agilent LC features, modules, and systems • Reduce Costs Access new Agilent features in non-Agilent software, without extra coding Page 48 June 27, 2013