NEW 1200 Infinity Series Scaling LC performance while ... · PDF file1220 Infinity LC. 1260 Infinity LC. 1290 Infinity LC. NEW 1200 Infinity Series Raising the Standard in HPLC Scaling

1220 Infinity LC

1260 Infinity LC

1290 Infinity LC

NEW 1200 Infinity Series Raising the Standard in HPLC

Scaling LC performance while keeping method transferability

–

a contradiction?

„UHPLC for Every Lab!“

Stefan SchuetteJune 18, 2010

Page 1

Presenter

Presentation Notes

Plain Orange (Pantone 144): 100% - RGB equivalent: (R) 243 / (G) 154 / (B) 0 Mid-tone Orange: 65% - RGB equivalent: (R) 251 / (G) 189 / (B) 104 Light Orange: 38% - RGB equivalent: (R) 253 / (G) 217 / (B) 169

Agenda

• Agilent introduces a new HPLC Standard

• Scaling Performance while maintaining method transferability

April 20102

Confidential

Agenda


• Scaling Performance while maintaining method transferability

April 20103

Confidential

Page 4

The NEW Agilent 1200 Infinity Series

1290 Infinity LC 1290 Infinity LC

1200 Series

1120 Compact LC

1260 Infinity LC

1220 Infinity LC

NEWNEW

EnhancedEnhanced

NEWNEW

Page 4

Page 5

The NEW Agilent 1200 Infinity Series

Low-Flow LC

Prep-Scale LC

newnew

newnew

newnew

newnew

newnew

newnew

enhancedenhanced

newnew

newnew

Presenter

Presentation Notes

Here you see complete overview of the new 1200 Infinity Series. The first thing that is evident: Based on a uniform platform, using common modules and the same parts in the production, you get from Agilent the broadest portfolio of liquid chromatography instrumentations of all manufacturers. This ensures lowest training for using and maintaining different Agilent LC systems and highest uptimes of instruments because of mature parts. You can find individual parts in a 1290 Infinity LC down to a 1220 Infinity LC. Almost all parts in a 1220 Infinity are common to a 1260 Infinity as well. A micro-flow sampler, a analytical flow sampler and a prep-flow sampler are using the same mechanical parts. This allows optimized production resulting in optimized quality and longest lifetime. Coming to the details. On the left you see the 1220 Infinity LCs, in the middle the 1260 Infinity LC that comprises of the low-flow, analytical flow and prep-flow systems and at the right you see the flagship of the series – the 1290 Infinity LC. The 1220 Infinity LCs come in three flavors – we will see them later. The analytical flow 1260 Infinity systems comprises of a Binary, Quaternary and Isocratic system as well as the just lately introduced Binary-SFC system developped together with Agilent‘s cooperation partner Aurora and also highly notable a new bio-inert quaternary LC system. So far no technical changes to the low-flow and prep-flow systems have been made, they were just incorporated to the 1260 Infinity LC.

Page 6

NEW Agilent 1200 Infinity Series Raising the Standard in HPLC – Highest Value for any Budget

1260Infinity LC

Modular600 bar80 Hz

2x or 10x UV Sensitivity100% HPLC Compatible

HPLC Price –

UHPLC Capability

1220Infinity LC

Integrated600 bar80 Hz

Most affordableHPLC and UHPLC

1260/1290 Technology

1290Infinity LC

Modular1200 bar160 Hz

Highest PerformanceHighest Flexibility

HPLC Service Costs

Presenter

Presentation Notes

Let‘s have a look to the key features of the new 1200 Infinity systems in the analytical flow rate range. [CLICK] First of all a short explanation: RRLC – Rapid Resolution LC – means the ability of a LC-system to make use of sub-two micron particel size columns to achieve faster - more rapid - and higher resolving analysis. This is also called UHPLC and has many other synonyms. The 1220 Infinity LC has an integrated design but also allows upgrading and combination with additional 1260 Infinity modules. It already gives you full RRLC-capability because it has a maximum system-pressure of 600 bar and a high speed detector to cope with very narrow peaks for fastes analysis. The most remarkable point is its price. Together with its easy of use it is an ideal workhorse for routine analysis where an increase of throughput is sought at an highly affordable price. It is the most afordable entry point to RRLC on the market. [CLICK] The 1260 Infinity LC is the evolution of the 1200 Series LC. Being modular and now offering 600bar system pressure throughout all analytical flow range systems. That means it is 100% compatible with standard HPLC analysis but has the power range to run any RRLC application. Besides standardizing on 600bar maximum pressure for all pumps, autosampler and valves all standard UV-detectors come now with 80 Hz data rate. The new 1260 Infinity DAD uses the technology of the 1290 Infinity DAD and gives a 10-fold sensitivity compared to other diode array detectors. And finally, all these new features that allow you to benefit from faster analysis, faster turnaround times and higher resolutions and sensitivities come at an HPLC price! [CLICK] The 3rd pillar of the 1200 Infinity Series is the already last year introduced flagship – the 1290 Infinity LC offering up to 1200 bar and 160 Hz detector speed for highest performance and highest flexibility in point of applications and any column dimensions. And also here we have a strong message for your budget: it comes with standard HPLC service costs – very important for your cost of ownership over many years of usage. On the following slide you see the power range of all 1200 Infinity Series LC in an overview. [CLICK]

Page 7

The new Agilent 1200 Infinity Series

0

200

400

600

800

1000

1200

0 2 4 6 8 10

All analytical columns

Method Transfer

Higher Resolution

Faster Analysis

More viscous solvents

Lower temperatures

Semi-prep

1290 Infinity LC

1260 Infinity Quaternary/Iso LC1220 Infinity LC

Pressure [bar]

Flow [mL/min]

1260 Infinity Binary LC

Presenter

Presentation Notes

With this power range you are able to use any analytical column, can do easy method transfer, have the power for high resolution and fastest analysis. With the 1290 Infinity LC even down to narrow bore columns with uncompromised performance. You can use more viscous solvents and lower temperatures without hitting the pressure limit of your conventional LC-system. The 1260 Quaternary and Isocratic LC as well as the 1220 Infinity LC give you even a flow rate range that is suitable for semi-preparative applications. But now let‘s see what the RRLC capabilities on a Quaternary, Binary or even Isocratic LC system could mean for your daily work... [CLICK]

Instrument Column Compatibility

STM-short: sub-2µm particles, 30 –

50mm lengthSTM-long: sub-2µm particles, 100 –

150mm lengthHPLC: 3 -

5 µm particles

1)

Because of delay volume.2)

Because of delay volume and pressure.3)

Because of pressure.

Column TypeParticle

UHPLC< 2um (STM)

UHPLCSuperficial Porous

HPLC3 –

5um

Column Length / mm Short: 30 –

50 Long: 100 –

150 30 -

150 50 -

300

CoIumn I.D. / mm 2.1 3.0 4.6 2.1 3.0 4.6 2.1 3.0 -

4.6 3.0 -

4.6

Max Pressure / bar 1200 600 600 1200 600 600 600 600 400

Agilent Column RRHD RRHT RRHT RRHD RRHT RRHT Poroshell Poroshell Various

1290B –

1200bar

1260B –

600bar 2)

1260Q/1220G –

600bar 1) 2) 1)

1100/1200 –

400bar 2) 2) 2) 2) 3) 3)

Any column of any dimension and particle can be used on 1220/1260/1290 Infinity LC‘s

1200 Infinity Series matches UHPLC columns pressure rating:

•

1290 Infinity LC: 1200bar + lowest delay volume (DV) for optimum support of 2.1 -

4.6mm ID STM (RRHD)

•

1260/1220 Infinity LC: 600bar + std DV for optimum support of 3 -

4.6mm ID STM (RRHT) & SP (Poroshell)

best

good

accept.

compr.

NEW: 1220 Infinity LC Isocratic and Gradient RRLC at most affordable Price

Most affordable access to 1260 Infinity Quality & Performance

•

Configurations: •

1220 Infinity manual isocratic system (upgradeable!)•

1220 Infinity manual binary gradient system (upgradeable!)•

1220 Infinity automated binary gradient system•

Productivity: 10x higher than conventional HPLC systems•

600 bar up to 5 mL/min (10 mL/min at 200 bar) •

80 Hz detector speed•

Technology and Quality: 1260 and 1290 Infinity inside!•

Degassing: Same Technology/Parts as 1290 Bin/1260 Quat degasser•

Pump: Same Technology/Parts as 1260 Quat Pump (2-channel)•

Autosampler: Same Technology/Parts as 1260 ALS (no cooling) •

VWD: Same Technology/Parts as 1260/1290 VWD•

Price: markedly below 1260 Infinity Isocratic / Quaternary Systems

Page 9

Presenter

Presentation Notes

The 1220 Infinity LCs are surely the most affordable LCs that allow access to Rapid Resolution LC. The 1220 Infinity LCs are available either as a manual injection, isocratic system, a manual injection, binary gradient or also as a automated injection, bianry gradient system. Also important to know, upgrades are easily possible! Its power-range is identical to the new 1260 Infinity Quaternary LC that is why it has full RRLC-capability. And it comes with uncompromised quality – many mechanical parts are identical to the 1260 Infinity systems, some parts can be even found in a 1290 Infinity LC. During production of these instruments you cannot say on a certain sub-assembly if it will end up in a 1220 or 1260 Infinity system! But the price is significantly lower than a 1260 Infinity LC. That is because many redundant parts in a 1260 Infinity LC that allows the flexibility of a modular system are saved in an integrated design like in the 1220 Infinity LC. For example, power supplies, electronics, covers and many more – quite logical, right?

NEW: 1260 Quaternary LC System Enhanced Quaternary UHPLC at Quaternary HPLC Price

Evolution of best-selling 1200/1100 Series Quat LC*

•

Productivity: 10x higher than 1200/1100 Quaternary HPLC•

600 bar up to 5 mL/min (10 mL/min at 200 bar) •

80 Hz Detector Speed

•

Sensitivity: Up to 10x higher than 1200/1100 Quaternary HPLC•

10x by new 1260 Infinity DAD with 6 cm flow cell (±0.6µAU)•

2x by new 1260 Infinity VWD and 1260 Infinity DAD with 1cm flow cell

•

Technology: Leverage of 1290 Infinity LC innovations•

Pump: New, build-in degassing unit and next generation inlet valves•

Detectors: Next generation VWD and DAD-technology

•

Price: Similar as 1200 Series Quat (400 bar, 20 Hz, ±6 µAU)

* ) > 60,000 systems sold!

Page 10

Presenter

Presentation Notes

The new 1260 Infinity Quaternary LC is the evolution of the best selling 1100/1200 Series Quaternary LC. Because of its new power range it allows the use of sub-2-micron columns and gives a productivity improvement up to 10-fold. But also the UV-sensitivity has significantly improved, actually it is the biggest step in sensitivity improvement ever! The new 1260 Infinity DAD allows up to a 10-fold increase in sensitivity! This gives you better peak detection, thus making integration easier. Higher resolution and higher sensitivity will give you much more confidence in your results – never miss a peak again! And again, here we have levered innovations from the 1290 Infinity LCs. The next generation inlet valves of the 1290 Infinity Pump are used here as well, giving a better performance. Also the integrated degassing unit – here of course a four channel unit – serves at higher performance levels, giving you less baseline disturbances. And as promised, coming at a price similar to the well known 400bar and 20Hz 1200 Series Quat-LC system.

NEW: 1260 Binary LC System Enhanced Binary RRLC Capability at HPLC Price

Successor of 1200 Series RRLC (1200 SL)

•

Productivity: •

Same as the highly successful 1200 Series RRLC –

for 2.1 mmID to 4.6 mmID columns, by scalable delay-volume

•

Sensitivity: •

10x higher than 1200 Series RRLC by new 1260 Infinity DAD •

Autosampler-Performance: •

<0.004 % carry-over and

100 µL injection by new 1260 Infinity High Performance Autosampler. 40 µL kit for delay volume reduction.

•

Technology: Leverage of 1290 Infinity LC innovations•

Pump: New next generation inlet valves for better performance•

Detectors: Next generation 1290 Infinity DAD and VWD technology•

HiP Autosampler: Same technology and parts as 1290 Autosampler•

Price: Markedly lower than 1200 Series RRLC

Page 11

Presenter

Presentation Notes

The 1260 Infinity Binary LC is the successor of the very successful 1200 Series Rapid Resolution LC that became now the standard for all modular binary LCs from Agilent – but with a significantly lower price compared to the 1200 Series RRLC. But it is not just a relabeling – also here you have many important enhancements. First, the new sensitivity level of the new 1260 Infinity DAD, also the improved data rate – 80Hz already as standard for the UV-detectors, 74 Hz for the new fluorescence detector and also 60Hz for the ELSD detector. Secondly, the high performance autosampler – this can of course be used as well for the other 1260 Infinity LCs – is improved and uses many parts of the 1290 Infinity autosampler. One important point should be mentioned regarding the pump – it has a scalable delay volume that makes it also very well suited for narrow-bore columns but of course not at the maximum performance level as the 1290 Infinity pump.

1290 Infinity LC More Affordable Access to the World‘s Best LC

NEW: UHPLC Productivity with HPLC Service Costs

•

New services pricing similar to 1200 Series, attractive contracts•

Based on –

proven robustness –

2x higher than 1100 at introduction–

Increased parts lifetime –

longer maintenance intervalls–

new design-for-supportability technology for faster maintenance, diagnostics and repair

NEW: More Flexibility, Performance and Affordability

•

New Multi-method/Walk-up and Method Development Solution•

New high-capacity solution for highest-throughput applications•

New lowest-carry over solution (<0.001%, <10ppm with 1290 FlexCube)•

More affordable entry-level configurations using same technology

12

Page 12

Presenter

Presentation Notes

Finally, the 1290 Infinity LC. Also here you get a couple of improvments. The biggest improvement will be to your budget – the service costs have been reduced on March 1st to a standard HPLC-service cost level! This is based on its proven robustness, increased parts lifetime, longer maintenance intervalls and new levels of design-for-supportability for faster maintenance and diagnostics. Technology-wise, it should be mentioned that the 1200 bar system pressure is of course available for all components, including a wide portfolio of valves for many sophisticated applications. For example, the 8-fold column selection for method development. Also remarkable, together with the recently released 1290 Infinity Flexible Cube the 1290 Infinity Autosampler achieves near zero carry-over performances. Typicall less than 10ppm, i.e. 0.001% carry-over after an injection of more than 1000 ng of chlorohexidin in UV detection, or after 50 ng injection with ultra high-sensitivity detection using an Agilent 6460 QQQ.

NEW: 1260 Infinity Diode Array Detector 2x or 10x higher Sensitivity at similar Price

13

NEW: Successor of 1200 DAD and DAD SL

•

Sensitivity: •

Same as 1290 Infinity DAD•

1-cm flow cell: 2x higher than 1200 DAD/1200 DAD SL•

6-cm flow cell: 10x higher than 1200 DAD/1200 DAD SL•

Technology: •

Same as 1290 Infinity DAD •

80Hz (instead of 160 Hz on 1290 DAD)•

4nm fixed slit (instead of variable slit on 1290 DAD)•

Price:•

Markedly lower than 1290 DAD and 1200 DAD SL•

Similar to 1200 DAD

Page 13

History of DAD Sensitivity Gain - The last 30 years

Detector Model (Intro Year)

Sensitivity Gain

(1988)(1986) (1995) (2006) (2010)

1260 / 1290 Infinity Diode Array Detector

Page 14

Page 15

Deuterium Lamp

Max-Light Cartridge Cell

Programmable

or fixed slit

1024 element

diode-array

GratingMirror

Optofluidic Waveguides: Max-Light Flow Cells Total-internal reflection in a non-coated fused silica fiber

1260 / 1290 Infinity DAD

11.4 x Higher Sensitivity 1290 Infinity DAD compare to 1200 Series DAD SL

July 8, 2010

Confidentiality Label16

1260/1290 DAD 60 mm

1200 DAD SL 10mm

Height [mAU] 28.876 4.938

Noise [mAU] 0.0098 0.0190

Signal/ noise 2944 259

Increase 11.4 x

min0 1 2 3 4 5

mAU

0

5

10

15

20

25

30

Columns: 150 x 4.6mm Zorbax SB C18, 5µm

Sample: Anthracene: 835 pg/µL

Mobile phase: A: Water, B: Acetonitrile Elution: isocratic 80 % B

Injection volume: 5 µL

Flow: 1.5 mL/min

DAD: 251/4nm, Ref= 450/80nm,

2.5Hz, slit width 4nm

1200 Series DAD / DAD SL

1260/1290 Infinity DAD

Presenter

Presentation Notes

Sample: Anthracene 835pg/µl Columns: 150x4.6mm Zorbax SB C18, 5µm Mobile phases: A= water, B= acetonitrile (20/80) Flow analytical pump: 1.5ml/min Column oven: 40°C on both sides DAD: 251/4nm, Ref= 450/80nm, PW>0.1min, 2.5Hz, slit width 4nm Injector: 5µl injection volume

11.4 x Higher Sensitivity 1290 Infinity DAD compared to 1200 Series VWD A/B

17

min0 1 2 3 4

mAU

0

5

10

15

20

25

30

Columns: 150 x 4.6mm Zorbax SB C18, 5µm

Sample: Anthracene: 835 pg/µL

Mobile phase: A: Water, B: Acetonitrile Elution: isocratic 80 % B

Injection volume: 5 µL

Flow: 1.5 mL/min

DAD: 251/4nm, Ref= 450/80nm,

2.5Hz, slit width 4nm

VWD: 251 nm, 2.5 Hz

1260/90 DAD 60 mm

1200 VWD10mm

Height [mAU] 28.876 4.800

Noise [mAU] 0.0098 0.0191

Signal/Noise 2944 253

Increase 11.6 x

1260/1290 Infinity DAD

1200 Series VWD A/B

NEW 1200 Infinity Series More workflow-automation and application-based solutions

18

AnalyticalSFC

GPCSEC

PreparativeLC & LC/MS

High-throughputLC & LC/MS

Bio-inert HPLC

NanoflowHPLC-Chip MS

Isocratic LC

Multi-method andMethod Development

Nano LC & Cap LC

New New

New New

Page 18

Presenter

Presentation Notes

I am almost at the end and would like to highlight some few general aspects of the 1200 Infinity Series. Here you see the many workflow solutions that are based on the 1200 Infinity series. The 1260 Infinity Binary SFC system allows HPLC-detection sensitivity in SFC and also comes with a very strong budget-message: by using standard-grade CO2 the operating costs are 10-15-times lower compared to other SFC-systems on the market that need high purity CO2 and still not achieve the detection limits of the new Agilent solution. The brand new 1260 Infinity bio-inert Quaternary LC sets also new levels of performance in analysing biological samples. With comes with the same power range and sensitivity level of the non-bio-inert 1260 Infinity Quaternary system. It uses a Titanium-made pump and completely metal free flow-paths. This is also available with an bio-inert fraction collector for semi-prep work. Finally, I like to mention also the multi-method and method development solution that allows either based on a 1260 or 1290 Infinity LC the automated selection between up to 8 columns and up to 26 solvents. .... [CLICK]

NEW 1260 Infinity Bio-inert HPLC Solution Infinitely Better for Bio-molecule Analysis

100% Bio-inert•

Metal-free sample flow path (instrument and column) •

Stainless steel–free solvent flow path •

Widest pH range (1-13, short-term 14)

10x sensitivity and 600bar power range•

Lowest LOD and highest resolution per time•

From lowest pressure up to highest pressure STM columns •

Unique resolution for protein applications

Bio-analysis and Bio-purification•

Analytical flow and

semi-prep fraction coll. up to 10 mL/min

Most complete offering for NBE Characterization •

Therapeutic protein analysis using SEC, IEX, peptide mapping

Page 19

New

NEW 1260 Infinity Analytical SFC System Infinitely Better for Supercritical Fluid Chromatography

Makes routine analytical SFC a reality!

Highest Analytical SFC Performance•

HPLC-like Sensitivity (10x over existing instruments)•

Precision and Dynamic Range >10,000 for accurate quantitation of 0.01 % level impurities

•

UHPLC-like speed on 1.8µm RRHT/RRHD columns

Lowest Operating Cost, Green Chemistry•

10 – 15x lower operating costs by compatibility with standard grade CO2 instead of liquid SFC grade CO2

•

Lowest solvent consumption and waste generation

Ease of use and Reliability•

Highest investment protection by modular, reversible design•

ChemStation control, data analysis and reporting •

Agilent warranty and service quality•

Single Vendor Solution –

Single Vendor Support

Page 20

New

Full module compatibility

•Mix 1220, 1260, 1290 Infinity LC with 1100 and 1200 Series –

and future systems !

Stepwise upgrade – now and in future:

•Exchange Detector only –

for higher sensitivity and detection speed.

•Exchange Pump and Autosampler only –

for ultra-high resolution LC on latest 1.8 µm rapid resolution column technology

•Exchange Column Compartment only –

to upgrade to a fully automated multi-method or method development solution supporting up to 8 columns and 24 solvents.

NEW: 1200 Infinity Series – Agilent Value Promise Infinitely Better Investment Protection

Page 21

Presenter

Presentation Notes

If you take the complete 1200 Infinity Series and the Agilent LC-Systems in general there are also important points to consider. First, within the 1200 Infinity Series all modules are mixable that means you can use a 1290 Infinity detecor on a 1260 Infinity system or vice versa. Use the 1290 Infinity Column Compartment for total valve flexibility in your 1260 Infinity stack. Add a special 1260 Infinity detecor to a 1220 Infinity system. All this is no problem! Within the 1200 Infinity Series many upgrade paths exist from simple systems to highly complex systems – as your requirements might grow over time. Also you can mix and match 1200 Infinity Series modules with older 1200 Series or even 1100 Series systems for a step-wise upgrade. Maybe just start with a more sensitive detector. That is an infinetly higher investment protection and was always the Agilent philsophy not to leave you alone with a once purchased Agilent system.

NEW: Instrument Control Framework (ICF) The Infinitely Better 3rd Party Connectivity Solution

Advantages for Customers •

Immediate Control –

Plug and Play (like windows printer driver)

•

Full and Robust Control –

all Features Supported by Agilent Drivers

•

Future Proof Control –

Backwards compatibility of Funtional Extension

Advantages for Application Software Suppliers•

Cost Savings –

Develop one Adapter instead of 50 Drivers, No more instrument purchase

•

Customer Satisfaction –

Immediate control, No Feature Missing

•

Risk Reduction –

Use tested Agilent Driver

NEW

Ada

pter

NEW

ICF

RC.NET LC Driver

RC.NET LC Driver

RC.NET LC Driver

3rp Party Supplier Agilent: 1100/ 1120/ 1200/ 1220/ 1260/ 1290

Page 22

Agenda


• Scaling performance while maintaining method transferability

April 201023

Confidential

Scaling Performance while maintaining Method TransferabilityWhy Important?

Performance

•

Sample Throughput, Productivity•

Sample Knowledge, Data Quality, Security

Method Transfer•

Run established, validated methods unchanged and get same results•

Match System of Client or Customer•

From R+D to QA•

From QA to R+D

April 201024

Confidential

How to achieve highest Peak Capacity at given Gradient Time and System Pressure?

•

t_g < 5min: 50 mm Column Length

•

5min < t_g < 30min: 100 mm Column Length

•

t_g > 30min: 150mm Column Length

June 06, 2005Page 25

By applying maximum Flow Rate (and Temperature)!

P. Petersson et al (AZ), J.Sep.Sci, 31, 2346-2357, 2008

D. Guillarme et al, Journal of Chromatography A, 1216 (2009) 3232–3243, 2009

26

1200 Infinity Series Power Range and 3-4.6mm, 1.8um TP columns

bar

ml/min

50 x 4.6 mm50 x 3.0 mm100 x 4.6 mm150 x 4.6 mm250 x 4.6 mm

H2 O/ACN, 40 °C

STM = Sub-2um SP = Superficially PorousTP = Totally Porous

Scaling Performance: 1260 Quat / 1220 Grad UHPLC Performance for EVERY LAB!

Resolution optimized3.0x100mm, 1.8µmResolution peak 5 = 7.16Run time 7 min0.9mL/min, 460bar

Speed and resolution optimized3.0x50mm, 1.8µmResolution (peak 5) = 4.79Run time 1.1min2.4ml/min, 540bar

Conventional4.6x150, 5µmResolution peak 5 = 4.20Run time 11 min

min0 2 4 6 8 10

min0 2 4 6 8 10

min0 2 4 6 8 10

10x productivity increase and

improved data quality!

Page 27

Presenter

Presentation Notes

A quick recap on sub-two-micron columns: if you reduce the particle size of your columns you get higher resolutions with the same column length – you have certainly already used this in your lab by going down from 5 µm particles to 3.5µm particles. If you reduce the particle size even more, another important effect takes place: You certainly know the van-deemter curve that, simply speaking, shows the dependency of the resolving power to the applied flow rate. With large particles the curve has a distinct minimum with a steep increases of the curve outside the minimum – that is, giving you lower resolutions at either very low or very high flow rates. Not so with very small particles – at higher flow rates the curve is almost flat and allows the use of high flow rates almost without loss of resolving power! That is the secret to achieve with so-called sub-two-micron columns high efficiencies and high flow rates, i.e. short run-times. Now, what does this mean for you, if you decide on a 1220 or 1260 Infinity LC? Have a look to the chromatograms generated on a 1260 Infinity Quaternary system. First, you see that you can run any conventional columns - even up to 300mm length - under conventional conditions without any problems – that is the top chromatogram. By using now a column with 1.8µm particles you can increase the resolution and reduce the run-time. And you have the flexibility to decide between highest resolution – shown in the middle chromatogram - or highest speed and still good resolution – shown in the bottom chromatogram. You might have noticed that the column diameter was reduced as well – this allows saving of solvents! Exactely the same results are available with the 1220 Infinity LCs. But let‘s have a closer look to the new systems and explore even more benefits for your daily work. [CLICK]

Scaling Performance: 1260 / 1220 InfinityUHPLC Performance for EVERY LAB!

4.6 x 150, 1.8um490 barN = 28669 R_S = 1.80 (+ 57%)S/N = 44

7 Impurities

All 7 Baseline Separated!

4.6 x 150, 5um93 barN = 7259R_S = 1.15S/N = 42

4 Impurities

2 Not Baseline Separated!

Customer Example Isocr. Impurity Method

Zoom of critical time range @ 7min

4.6 x 150, 3.5um165 barN = 14862R_S = 1.37S/N = 50

7 Impurities

6 Not Baseline Separated!

Presenter

Presentation Notes

Resolution gains according to theory prove the ability of the 1200 RRLC system to use standard bore instead of narrow bore RRHT columns for highest resolution RRLC applications without sacrificing chromatographic efficiency by thermal mismatch of mobile phase and column. (Such a thermal mismatch would cause band broadening due to temperature gradients across the column radius.) This disproves the Waters Statement: “The use of 4.6mm ID columns for UPLC is not practical due to frictional heating generated within the column due to the flow rate and the inability to sufficiently dissipate the heat due to limited surface area to volume ratio therefore….. real UPLC uses 2.1mm ID columns”). See also next slides. Note: The advantage of using standard bore RRHT columns is that these can be used on the standard delay volume configuration, which offers uncompromised compatibility with conventional HPLC methods – see Benefit 3.

Different Aspects of Method Transfer

From Instrument to Instrument -> with method/instrument change: Isocratic Hold / Pre-injection, Vol. Modification -> available tools w/o Method or Instrument change : none so far

From column dimension to column dimension (e.g. 4.6 -> 2.1, 50 mm -> 100 mm)-> recalculate flow rates, recalculate gradient times, adjust connection capillaries and flow cells, due to delay volume results may vary, method might need revalidation-> available tools: Agilent Method translator

From particle size to particle size (HPLC -> UHPLC)-> often associated with a change of instruments due to constraints in max. pressure, extra-column band broadening etc.. Some fine-tuning of method usually required due to frictional heating effects-> available tools : Agilent Method translator, Third Party Method development SW (ACD Labs, ChromSword etc.)

From one eluent type or phase chemistry to an other-> see Method development-> available tools : Method development SW (Chromsword, ACD Labs,

etc.)

Method Transferability – how is this possible? Run Method and Get Same Result?

1200 Series

1100 Series

1220 Infinity

1260 Infinity

1290 Infinity

?

Page 30

Presenter

Presentation Notes

I said a couple of times that we will have 100% method compatibility with your existing systems. How is this possible? Run a method on a 1200 or even 1100 Series system and get the same result on a new 1260 Infinity system.

Method Transferability: 1260 Quat / 1220 Grad 100% HPLC Methods Compatibility – Riskless Replacement

1260 / 1220 Infinity LC Quat / Gradient system

1100 / 1200 Series Quaternary system

Column: 4.6 x 150 mm, 5 µmGradient:

0 min 20 % B10min 95 %

Flow rate:

1 mL/min

min0 2 4 6 8 10

Run existing conventional HPLC methods and

get same result !

Page 31

Presenter

Presentation Notes

Here you see the comparison of a conventional method run on a 1100/1200 Series Quaternary LC (blue trace) and than transferred directly without any changes like adding isocratic holds or additional tubing to the system to a 1260 Infinity (red trace). You can‘t see any differences, right? That is true method compatibility for riskless replacement! Exactely the same results are achievable with a 1220 Infinity LC! ... But there is also a second way to achieve 100% method transfer – actually a very sophisticated one. [CLICK]

1100/1200 Series

HPLC

1260 InfinityQuaternary LC

1200 Series RRLC – Std.

1260 InfinityBinary LC

Max Flow Rate 5 mL/min 5 mL/min 5mL/min 5 mL/min

Delay Volume 600‐900 µL 600‐800 µL 600‐800 µL* 600‐800 µL*

Capillary ID 0.17mm 0.17mm 0.17mm 0.17mm

Disp. Vol. w/o cell 15µl 15µl 15µl 15µl

Injection Principle Variable Loop Variable Loop Variable Loop Variable Loop

Inj.Volume –

Std/Ext. 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL

Area RSD <0.25 % <0.25 % <0.25 % <0.25 %

Oven Design A A A A

Column Length 300 mm 300 mm 300 mm 300 mm

* Smaller delay volumes possible

Seamless Method Transfer 1260/1220 Infinity By unchanged critical specification

Page 32

Optimized for 3 –

4.6mm ID Columns

Optimized for 3 –

4.6mm ID Columns

Presenter

Presentation Notes

Well, there are two ways to achieve a seamless method transfer. First, you have really comparable systems. With the 1200 Infinity Series this is possible by an evolutionary approach to increase the performance. You can see in the table that the new 1260 Infinity LC come with comparable designs, but of course improved to achieve the new performances levels. We have the same flow rate on the pumps as the previous systems; so, no fear to run into a flow rate limitation. The oven design and the injection principle hasn‘t changed - also here no risks. Having a different column oven for example could mean you have inside the column different temperatures based on the heating system used. You all know these separations that react very sensitive on temperature changes, right? Not so with the Agilent 1260 Infinity LCs. The only tiny variation was made on the delay volume of the system. The 1260 Infinity pumps have a slightly reduced standard delay-volume. But with flow rates as typical in conventional HPLC you will hardly notice this at all. The optionally delay volume reduction of the 1260 binary pump for using 2.1mmID columns is of course a different story. See on the next slide how this results in transferring methods from a conventional system to a new 1200 Infinity Series LC. [CLICK]

33

Agilent 1290 Infinity LC Power Range and 2.1mm, 1.8um TP Columns

bar

ml/min

50 x 2.1 mm100 x 2.1 mm150 x 2.1 mm250 x 2.1 mm

0.45 mL/min

H2 O/ACN, 40 °C

ZORBAX RRHT SB-C182.1 x 50mmH2 O/ACN2.4ml/minT=95 °C560bar


1100/1200 Series

HPLC

1260 InfinityQuaternary LC

1200 Series RRLC – Std.




Max Flow Rate 5 mL/min 5 mL/min 5mL/min 5 mL/min 5 mL/min 5 mL/min

Delay Volume 600‐900 µL 600‐800 µL 600‐800 µL* 600‐800 µL* 120 µL** 10 – 110 µL*

Capillary ID 0.17mm 0.17mm 0.17mm 0.17mm 0.12mm** 0.12mm*

Disp. Vol. w/o cell 15µl 15µl 15µl 15µl 7.5µL** 7.5µL*

Injection Principle Variable Loop Variable Loop Variable Loop Variable Loop Variable Loop Variable Loop

Inj.Vol. –

Std/Ext. 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 µL

Area RSD <0.25 % <0.25 % <0.25 % <0.25 % <0.25 % <0.25 %

Oven Design A A A A A A

Column Length 300 mm 300 mm 300 mm 300 mm 300 mm 300 mm

* Smaller delay volumes possible

Seamless Method Transfer What if critical specification change ...?

Page 34

* Optimized for 2.1 -

4.6mm ID** Optimized for 2.1mm ID

Presenter

Presentation Notes

Well, there are two ways to achieve a seamless method transfer. First, you have really comparable systems. With the 1200 Infinity Series this is possible by an evolutionary approach to increase the performance. You can see in the table that the new 1260 Infinity LC come with comparable designs, but of course improved to achieve the new performances levels. We have the same flow rate on the pumps as the previous systems; so, no fear to run into a flow rate limitation. The oven design and the injection principle hasn‘t changed - also here no risks. Having a different column oven for example could mean you have inside the column different temperatures based on the heating system used. You all know these separations that react very sensitive on temperature changes, right? Not so with the Agilent 1260 Infinity LCs. The only tiny variation was made on the delay volume of the system. The 1260 Infinity pumps have a slightly reduced standard delay-volume. But with flow rates as typical in conventional HPLC you will hardly notice this at all. The optionally delay volume reduction of the 1260 binary pump for using 2.1mmID columns is of course a different story. See on the next slide how this results in transferring methods from a conventional system to a new 1200 Infinity Series LC. [CLICK]

Definition of Delay / Dwell / Transition VolumeScatter Plot

Delay volume (ml)0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8

0

10

20

30

40

50

60

70

80

90

Delay Volume @ 10% of step

Transition Volume

10% of step height

90% of step height

Dwell volume @ 50 % of step

Consequencs of different Delay Volumes 1200 RRLC versus 1290 Infinity System

36

Line Chart

Volume [ml]

20

100

190

280

360

450

530

620

700

203

10015

19027

27038

36050

44062

53074

61086

70097

0 0.5 1 1.5 2 2.5 3 3.5 4 4.5

%organ

ic

modifie

r

Programmed gradient

1290 Infinity System

1200 RRLC / SL System

% o

rgan

ic m

odifi

er

Method Transferability: 1290 Infinity LC Consequences of a lowest delay volume system

Lowest delay volume, accuray and precision of 1290 Infinity pump allows to match any other gradient behavior!

Page 37

time

%B

Presenter

Presentation Notes

If you have such a superior pump as the 1290 Infinity pump you can actually match the behavior of any other system. The secret lies in the very low delay volume of the 1290 Infinity pump and its ability to form gradients extremely precise. Let me explain the figure shown here. You see the result of a step-gradient monitored in the UV-signal by using a tracer in the one solvent. You will never get exactely what you have programmed in the gradient method because you always have some delay volumes in your system that are filled with the starting composition. When the programmed composition delivered by the pump, travels through the system it will be diluted and you get a different composition This will generate the true gradient that is relevant for the separation. In Blue you see the result using a standard 1200 Series RRLC system. You get a shift of the composition change in the time axis until the composition change of the programmed step arrives at the detector and you also get a slope because of the mixing behavior. In green you see the same step gradient achieved with the 1290 Infinity pump. This is mutch closer to the programmed step, actually no pump on the market gets that close! Additionally you see what happens if the additional delay volumes of the 1200 Series RRLC system are added to the 1290 Infinity system – you get the identical behavior! That means you get the identical chromatographic results because the sample experiences the same true mobile-phase composition.

Page 38

Method Transferability: 1290 Infinity LC Today‘s Bridging Approaches

1. Add physical Volume (plumbing solution)

2. Programmed Isocratic Hold *

1290 + 1000ul dwell vol.

1100/1200 Quat Pump

1200 RRLC

1290 + 900ul hold

Page 38

Presenter

Presentation Notes

Until the System Emualting Technology is available also bridging solutions can give good results. As you can see here adding physical volume – the upper part - or isocratic holds – the lower part – already give very good results. But the results using the coming system emulation-technology will take it to the next level!

Programmed Isocratic Hold Limitations

0

10

20

30

40

50

60

70

80

90

100

0 2 4 6 8 10 12 14

1290 Infinity

1200 SL

0

10

20

30

40

50

60

70

80

90

100

0 2 4 6 8 10 12 14

1290 Infinity

1200 SL

0

2

4

6

8

10

12

14

16

18

20

3 4 5

1290 Infinity

1200 SL

80

82

84

86

88

90

92

94

96

98

100

11 11.5 12 12.5 13

1290 Infinity

1200 SL

Gradient shifted by isocratic holdOriginal gradient response traces

Method Transfer Low to Large DV System Elements to consider

Gradients are delayed

AND “bent”

by transition volume

transition

delay

Composition accuracy:-

e.g. quaternary vs. binary pump concept-

systematic offset

offset

characterization

3.3.

2.2.

1.1.

How to overcome Limitations?

Fa

Fb

TT-a Delay/Transition a

TT-a Delay/Transition b

How to turn Delay/Transition b

into Delay/Transitition a?

Execution Conceptinput

output

TT-a

TT-b

Method

Ha-b

Fb

temporaryFb

Fa

F1290

F1200= =

No Change on Method / TT or Instrument !

Method Transferability: 1290 Infinity LC System Emulation Technology (planned for 2011)*

Ha-bEmulated System A

Agilent 1100 Series

Agilent 1200 Series RRLC

Select System to be emulated by a simple mouse click:Select System to be emulated by a simple mouse click:

Agilent 1260 Infinity LC

Agilent 1220 Infinity LC

Non-Agilent HPLC / UHPLC

ConceptConcept

* Existing 1290 Infinity will be upgradeable Page 43

Presenter

Presentation Notes

The capability of the 1290 Infinity pump allows a unique System Emulation Technology that will be available next year –of course, existing systems are upgradable since it is no technical change of the pump, just software. The concept is to use an emulating function that changes a gradient generated by the 1290 Infinity pump to match exactely the gradient – the true gradient resulting from the programmed one – of any other system. If you think back to the previouse slide it is clear what needs to be done: add the additional time at the beginning but then also add a slope to the gradient such that the true gradient of the 1290 Infinity pump will exatly match the true gradient of the emulated system. Finally, that means you can select any other LC system to be emulated by a simple mouse-click!

NEW 1220 and 1260 Infinity LC •

New HPLC Standard –

10x Productivity and 10x Sensitivity•

100% HPLC Compatible –

Riskless Replacement•

HPLC Price –

UHPLC Performance

Enhanced 1290 Infinity LC•

Highest Performance and Flexibility –

by any measure•

New: Highest UHPLC productivity –

HPLC Service Costs•

New: highest-productivity solutions –

and more affordable entry-level configurations

NEW 1200 Infinity Series•

Continuum of UHPLC Solutions –

starting at < 20k$•

Scalable and Compatible –

across entire 1200 Infinity Series and 1100/1200 Series•

Common technology and quality –

for highest robustness and reliability•

Method transferability and compatibilty –

within entire series and 1100/1200 Series•

New workflow automation and application-based solutions –

incl. Bio-inert HPLC and SFC•

New ICF –

for seamless integration into 3rd party software

NEW Agilent 1200 Infinity Series – Summary

44

http://www.agilent.com/chem/infinity

Page 44

http://www.agilent.com/about/newsroom/lsca/

Agilent´s Analytical LC Portfolio Compatible and Scaleable across entire Series

Page 45

• Run 1100/1200 Methods on 1260/1290 Infinity LC

• Upgrade 1100/1200 with 1260/1290 Modules

• Full module and methods compatibility

• Highest investment protection

Page 45

Introduction

46

Modern UHPLC instruments are optimized for fast separations in small diameter columns.

Delay volumes have been reduced by a factor of 5-10 compared to conventional HPLC instrumentation.

Due to differences in pump and mixer design differences in gradient formation exist between different types of HPLC instruments

Approaches have been evaluated to overcome these problems and a novel solution is proposed to provide method compatibility between HPLC and UHPLC is proposed.

Comparison of Gradient Runs on Different Instruments

Column: Zorbax SB-C18, 2,1x50 mm, 1,8µMobile phase: A = water / B = acetonotrileSig: 254,10nm; Ref: 360,100nm

Temperatue: 40°CInj.vol.: 1µlFlow rate: 0,5 ml/min

Sample: Sigma

1290 Infinity

min0 1 2 3 4 5

mAU

020406080

0.25

6 1.62

9

1.98

0

2.40

6

2.80

42.

887

3.16

0

3.35

2

4.07

1

1200 SL Binary Pump

min0 1 2 3 4 5

mAU

020406080

0.30

4

2.52

4

3.18

0

3.61

6

4.01

94.

104

4.38

1

4.54

9

5.32

7

Gradient: 5%B to 95%B, TG: 5 min

min0 1 2 3 4 5

%

020406080

Page 48

Current Common Solutions

Programmed isocratic holdadd physical Volume (plumbing solution)

“Plumbing solution”: Comparison 1100/1200 Quaternary Pump vs 1290 Infinity



Sample: RRLC check out

1290 Infinity

1290 + 1000µl dwell

volume *

1100/1200 Quaternary Pump


min0 2 4 6 8 10

mAU

0

40

80

120

3.92

8

5.51

6

6.62

0

7.47

27.

783

8.23

4

8.92

9

9.55

6

10.1

16

min0 2 4 6 8 10

mAU

0

40

80

120

4.49

5

6.10

2

7.21

4

8.06

6

8.37

7

8.82

7

9.52

1

10.1

46

10.7

05

min0 2 4 6 8 10

mAU

0

40

80

120

1.19

4

4.45

4

6.05

6

7.17

7

8.04

2

8.35

8

8.81

6

9.52

1

10.1

57

10.7

28

min0 2 4 6 8 10

%

020406080

100

*experimentally obtained dwell volume value

“Plumbing solution”: Comparison 1100/1200 Binary Pump vs 1290 Infinity



Sample: RRLC check out

1290 Infinity

1290 + 1000µl dwell volume *

1100/1200 Binary Pump


min0 2 4 6 8 10

mAU

0

40

80

120

3.92

8

5.51

8

6.62

1

7.47

4

7.78

5

8.23

6

8.93

2

9.56

0

10.1

20

min0 2 4 6 8 10

mAU

0

40

80

120

4.49

4

6.10

2

7.21

5

8.06

6

8.37

7

8.82

8

9.52

2

10.1

46

10.7

03

min0 2 4 6 8 10

mAU

0

40

80

120

4.53

6

6.14

0

7.24

5

8.08

6

8.39

0

8.83

7

9.52

1

10.1

36

10.6

89

min0 2 4 6 8 10

%

020406080

100


“Plumbing solution”: Comparison 1100/1200 Binary Pump vs 1290 Infinity



Sample: Sigma

1290 Infinity

1290 + 1000µl dwell volume*

1100/1200 Binary Pump


min0 1 2 3 4 5 6 7 8 9

mAU

020406080

1.05

51.

143

4.19

4

4.60

9

5.43

5

6.23

06.

475

6.95

7

7.61

5

8.85

6

min0 1 2 3 4 5 6 7 8 9

mAU

020406080

1.05

4 4.73

2

5.19

9

6.03

4

6.83

07.

077

7.55

3

8.20

0

9.44

8

min0 1 2 3 4 5 6 7 8 9

mAU

020406080

1.10

71.

331

4.76

8

5.22

6

6.05

2

6.84

37.

088

7.56

1

8.20

9 9.43

2

min0 2 3 4 5 6 7 8 9

%

020406080


Presenter

Presentation Notes

The following slides with show the results achieved in the comparison of the original method (bottom), the method result on the 1290 (top) and on the 1290 after the modification and optimizing of the modification (middle). All conditions are mentioned esp. gradient, gradient length, column and sample.

“Plumbing solution”: Comparison 1200 SL Binary Pump vs 1290 Infinity



Sample: Sigma

1290 Infinity

1290 + 800µl dwell

volume *

1200 SL Binary Pump


min0 1 2 3 4 5

mAU

020406080

0.25

6 1.62

9

1.98

0

2.40

6

2.80

42.

887

3.16

0

3.35

2

4.07

1

min0 1 2 3 4 5

mAU

020406080

0.26

2 2.53

8

3.20

9

3.67

6

4.09

24.

177

4.45

3

4.62

2

5.36

2

min0 1 2 3 4 5

mAU

020406080

0.30

4

2.52

4

3.18

0

3.61

6

4.01

94.

104

4.38

1

4.54

9

5.32

7

min0 1 2 3 4 5

%

020406080


Isocratic Hold: Comparison 1200 SL / 1290 Infinity 5 min Gradient @ 1 ml/min

1290 + 860 µl Delay volume

1200 SL + Mixer + damper



Column: Zorbax EC-C18, 4.6x50 mm, 1,8µMobile phase: A = water / B = acetonotrileGradient 5 –

95 % B in 5 minSample: RRLC checkout sample

Isocratic Hold: Comparison 1200 SL / 1290 Infinity 10 min Gradient @ 1 ml/min (Alkylphenones)













95 % B in 10 min













Relative Difference in Retention Time between 1200 SL and 1290 Infinity with different programmed delays

Scatter Plot

Peak Number

Stop Flo w - 1

tg - 5

Stop Flo w - 2

tg - 5

Sto p Flo w - 1

tg - 10

Sto p Flo w - 2

tg - 10

Sto pFlo w - 1

tg - 20

Sto pFlo w - 2

tg - 20

2 4 6 8 10 2 4 6 8 10 2 4 6 8 10

-2

-1

0

1

2

-2

-1

0

1

2



Difference in Resolution of Adjacent Peaks

Scatter Plot

Peak Number

Stop Flo w - 1

tg - 5

Stop Flo w - 2

tg - 5

Sto p Flo w - 1

tg - 10

Sto p Flo w - 2

tg - 10

Sto pFlo w - 1

tg - 20

Sto pFlo w - 2

tg - 20

2 4 6 8 10 2 4 6 8 10 2 4 6 8 10

-5

-4-3

-2-1

01

23

-5

-4-3

-2-1

01

23



1290 Infinity Compatibility / Emulation ModeHypothesis:

Any instrument with a low delay volume (e.g. 1290 Infinity) can emulate an instrument with a larger delay volume. e.g.Agilent 1100/1200 / 1200 SL <-> 1290 Infinity w small/large Jet Weaver

Use Cases:Backward compatibility :Methods from larger delay volume systems can be run unchanged on the 1290 Infinity in emulation mode with same results

Method development:1290 Infinity in emulation mode can be used to develop methods for other systems

Possible implementationDetermine the transfer functions between 1290 and the system to be emulatedImplement an Algorithm that calculates gradient shape for emulation mode (FW/SW)Internally execute a modified timetable to emulate stating system

Execution Concept I

Fa

Fb

TT-a Delay/Transition a

TT-a Delay/Transition b

Execution Conceptinput

output

TT-a

TT-b

Method

Ha-b

Fb

temporaryFb

Fa

F1290

F1200= =

No Change on Method or Instrument !

Execution Concept I

Fa

Fb

x Transfer Function (complex) =input output

TTBL-a step response

= Transfer Function (complex)

output

inputFa=

Possible Workflow Schematics

CFR-21part11IVDPharma GMPFDA

Company (custom)guidelines

Application needs

System Suitabilityin GMP-environment

passed?no

yes

Rare case

-

Execute as e.g. 1050

-

Custom settings

-

Execute as e.g. 1200-SL

- - - -

documented Audit trail

allowed to use

SOP,Method

deviations

“e.g.” or equivalent equipment

“critical pair”performance specification

Details = nice- to-know

Run-sheetData filescomments

Conclusions

65

Due to minimized delay volumes in modern UHPLC instrumention, methods developed on standard HPLC instruments will not give the same results in retention time and resolution when run on an UHPLC system

Adding physical volumes „plumbing solution“

can lead to more comparable results but is not user friendly and requires exact knowledge of the difference in system delay volumes.

Adding an isocratic hold tho the method also does not always yield the desired result as it can correct for delay in gradient start but not for gradient shape.

A concept is proposed that allows to perform a SW/FW driven emulation of a variety of instruments on the Agilent 1290 Infinity system.

Systems can be pre-characterized but also tools for characterization of unknown systems an be provided.

1200 Rapid Resolution System Agilent Restricted

December 20, 2005Page 66

1200 Series TCC SL Dispersion: New 1.6µl-

versus standard 3µl-Heater

min1 1.5 2 2.5 3

mAU

0

50

100

150

200

250

1.235

1.358

3.227

1.216

1.339

3.181

1.6 µl Heater•

Sample:

Isocratic Standard Sample•

Flow rate:

0.5ml/min•

Eluent: Water/ACN = 30/70•

Column:

2.1x100mm, RRHT SB-C18 3 µl Heater

Presenter

Presentation Notes

selfexplanatory



1200 Series TCC SL Dispersion: New 1.6µl-

vs. standard 3µl-HeatEx.

Flow rate Evaluated Peak 1.6µl HeatEx. 50°C

3µl HeatEx.50°C

1.5ml/min Rs of peak 2 3.19 2.73

1.5ml/min PW 5sigma of peak 4 in min 0.031 0.033

1ml/min Rs of peak 2 3.96 (+26%) 3.15

1ml/min PW 5sigma of peak 4 in min 0.067 (-13 %) 0.076

0.5ml/min Rs of peak 2 3.29 2.86

0.5ml/min PW 5sigma of peak 4 in min 0.042 0.044

0.2ml/min Rs of peak 2 3.51 3.140.2ml/min PW 5sigma of peak 4 in min 0.192 0.197

Presenter

Presentation Notes

This table shows the analysis from the previous slide obtained at different flow rates and each one quantitatively evaluated for peak resolution and peak width. The resolution and peak width data obtained with the low dispersion heater are for each flow rate better. The oval highlights the comparison at a flow rate of 1.0 ml/min: resolution is by 26% and peak width by 13% better!



1200 Series TCC SL – ACR 2Ps/10Pt Valve and Heaters/Cooler for Alternating Column Regeneration

U-shaped post-column cooler 1.5 µl

2-position/10-port micro-valve SL

Two columns in alternate usage

Heater for column 1

Heater for column 2



Benefit 1 – Fastest LC Pharma Impurity Method –

Pat Sandra & Frank David, RIC

•

4.6 x 50mm RRHT SB-C18•

35% ACN at 0.6 min to 95% ACN at 2.5 min

•

3 mL/min, 40°C (450 bar)•

DAD SL: 13 µL cell –

40 Hz

Scaling Performance: 1260 BinaryUHPLC Performance for EVERY LAB!

min0 5 10 15 20

mAU

-1

-0.5

0

0.5

1

1.5

2

2.5

DAD1 A, Sig=254,4 Ref =of f (051119A\SIG10003.D) Column:150x4.6 mm

5µm

Pressure: 93 bar

N: 8213

Height: 1.25 mAU

S/N: 42.3

Rs

= 1.15

tr

= 14.9 min (1st

epimer)

Nptp

: 2.4 � 10-2 mAU

4.6 x 150, 5um93 barN = 7259R_S = 1.15S/N = 42 Height = 1.25Noise = 24uAU

Column:150 x 4.6 mm 1.8µm

Pressure: 490 bar

N: 28669

Height:1.78

S/N: 43.6

Rs = 1.80

tr = 17.2 (1st epimer)

Nptp: 3 10-2

min10

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5

3

DAD1 A, Sig=254,4 Ref=off (051007D\LC_X000001.D)


Pressure: 490 bar

N: 28669

Height:1.78

S/N: 43.6

Rs = 1.80


Nptp: 3 10-2

min10

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5

3


Column:150x4.6 mm

3.5µm

Pressure: 165 bar

N: 14862

Height:1,34 mAU

S/N: 50.7

Rs = 1.37

tr = 15.3 min (1st epimer)

Nptp: 2 10-2 mAU

min0 5 10 15 20

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5

DAD1 A, Sig=254,4 Ref=off (051004D\LC_J0002.D)

Column:150x4.6 mm

3.5µm

Pressure: 165 bar

N: 14862

Height:1,34 mAU

S/N: 50.7

Rs = 1.37


Nptp: 2 10-2 mAU

min0 5 10 15 20

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5



Pressure: 490 bar

N: 28669

Height:1.78

S/N: 43.6

Rs = 1.80


Nptp: 3 10-2

min10

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5

3



Pressure: 490 bar

N: 28669

Height:1.78

S/N: 43.6

Rs = 1.80


Nptp: 3 10-2

min10

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5

3


Column:150x4.6 mm

3.5µm

Pressure: 165 bar

N: 14862

Height:1,34 mAU

S/N: 50.7

Rs = 1.37


Nptp: 2 10-2 mAU

min0 5 10 15 20

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5


Column:150x4.6 mm

3.5µm

Pressure: 165 bar

N: 14862

Height:1,34 mAU

S/N: 50.7

Rs = 1.37


Nptp: 2 10-2 mAU

min0 5 10 15 20

Norm.

-1

-0.5

0

0.5

1

1.5

2

2.5


4.6 x 150, 3.5um165 barN = 14862R_S = 1.37S/N = 50 Height = 1.34Noise = 20uAU

4.6 x 150, 1.8um490 barN = 28669 R_S = 1.80 (+57%)S/N = 44 Height = 1.80Noise = 30uAU

1/dp

0.2 0.3 0.4 0.5 0.6

N

5000

10000

15000

20000

25000

30000

pdN 1

Presenter

Presentation Notes

Resolution gains according to theory prove the ability of the 1200 RRLC system to use standard bore instead of narrow bore RRHT columns for highest resolution RRLC applications without sacrificing chromatographic efficiency by thermal mismatch of mobile phase and column. (Such a thermal mismatch would cause band broadening due to temperature gradients across the column radius.) This disproves a competitor stetement: “The use of 4.6mm ID columns for UPLC is not practical due to frictional heating generated within the column due to the flow rate and the inability to sufficiently dissipate the heat due to limited surface area to volume ratio therefore….. real UPLC uses 2.1mm ID columns”). See also next slides. Note: The advantage of using standard bore RRHT columns is that these can be used on the standard delay volume configuration, which offers uncompromised compatibility with conventional HPLC methods – see Benefit 3.



Benefit 3 – The Most Flexible LC in the World!

min0 1 2 3 4 5 6 7

mAU

0

10

20

30

40

50

60

70

80

1100 Binary

1200 RRLC •

Low delay volume configuration (120µl)•

RT-Shifts up to 25%, difficult Peak tracking

•

Limited compatibility with HPLC methods

•

Typically requires method re-validation

-20% RT !

-25% RT !

Support of RRLC and

HPLC on Std and

Narrow bore columns1.

The 600-800µl standard delay volume configuration allows to run both standard bore RRLC and conventional HPLC methods without need of re-validation.

2.

The 120µl low delay volume configuration provides uncompromised compatibility with narrow bore LC and LC/MS applications in both RRLC and HPLC.

Presenter

Presentation Notes

The slides demonstrates that the low delay volume configuration offers only limited compatibility with conventional HPLC methods, because of the strong retention time shifts of up to 25%, which makes peak tracking difficult and typically requires re-validation of the method!



Competition – Waters Acquity + Alliance Application Range –

Universal vs. two

Expert Systems

1200 RRLC Acquity Alliance

RRLC – Narrow YES YES NO(6)

RRLC – Std YES NO(1) Limited(5)

HPLC – Narrow YES NO(2) Limited(4)

HPLC – Std YES NO(3) YES

(1)

Max flow rate of 2ml/min(2)

Max Column length of 200mm(3)

Max Column length of 200mm and

no std delay volume configuration => RT shifts of up to 30% => re-validation required

(4)

Delay volume too large (~1ml)(5)

No 1.7um column with 3 –

4.6mm ID, 400bar/60C limits

(6)

Del. vol. too large, 400bar/60C limits

Presenter

Presentation Notes

Three mayor features, which are not available on Acquity, turn the 1200 RRLC into a universal LC for the analytical flow rate range: Flow rates up to 5ml/min (for compatibility with standard bore RRLC/UFLC) Room for columns up to 300mm length (for compatibility with HPLC) Configurable delay volume (for compatibility with narrow and standard bore LC) Standard configuration with 600-800ul delay volume allows standard bore RRLC and HPLC) In contrast to the 1200 RRLC Acquity is a dedicated and inflexible narrow bore system only. I.e. it cannot offer the same investment protection like the 1200 RRLC, because of its limited compatibility with standard HPLC applications. Bottom Line: Waters customers need two systems – Acquity for UPLC (RRLC) and Alliance for HPLC, whereas Agilent customers need the 1200 RRLC only to run both RRLC and HPLC applications.



min0 0.2 0.4 0.6 0.8 1

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500

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700

0.209

0.370

0.505

0.620

0.664 0.7

25

0.821

0.908

0.986

1200 RRLC System Waters AcquityAnal. Time 0.949 min 0.914 minResolution 3.69 3.50Precision 0.025 –

0.094 %RSD 0.18 –

0.30 %RSDPressure 440 bar 700 bar (+60%)

Sample: Phenones Test MixColumns:

•

1200 RRLC: SB-C18, 2.1x50 RRHT •

Waters: BEH, 2.1x50Flow Rate: 1ml/minGradient: 35-95% ACN in 0.9minTemperature: 50°Injection volume: 1µlInjection Technique: ADVR, OI, MCOWL: 245nmData Rate: 80Hz

Competition – Waters Acquity 2.1mm Speed-Benchmark Method

1200 RRLC: Better Performance at significant lower pressure !!

Presenter

Presentation Notes

Due to system pressure optimization the 1200 RRLC system allows for better performance at significant lower pressure: Acquity columns generate approx. 25-30% higher pressure than Zorbax RRHT columns of same dimension. Reason: Zorbax RRHT columns use a patented particle size distribution optimization technology that reduces pressure significantly without giving up efficiency Acquity LC system itself generates much more pressure than the 1200 RRLC system. The higher the flow rate the higher the pressure difference. In the given example (1ml/min) Acquity generates approx. 30-35% higher pressure then the 1200 RRLC. Reason: Acquity needs longer and smaller ID connecting capillaries than the 1200 RRLC system. Note: The low column and system pressure of the 1200 RRLC system allows to run standard bore RRHT columns without loss in efficiency by thermal mismatch / band broadening.



Product Overview – 1200 RRLC System Configurations

System Configurator*

Column dimension

Flow Rate

Gradient

Eluent / Modifier

Temperature

Pump delay volume

Connecting Capillaries

Heat Exchangers

Flow Cell

Highest Speed, Res, or best Compromise?

*Available by February

Be Prepared !!

Presenter

Presentation Notes

Different analytical conditions may require different system setups in terms of pump delay volume, connecting capillaries, heat exchangers and flow cell. To find the optimum configuration for a certain analytical problem a so-called System Configurator will be provided by February, which will help the sales and service organization and customers to find a setup that best suits their needs. Most probably there will be 2 - 4 start configurations. For all start configurations a check-procedure will be available. To run the check-out a check-out column is needed that comes with the Binary Pump SL. Customers can choose between three check-out columns: 4.6 x 50 XDB-C18, RRHT (P/N: 927975-902) 3.0 x 50 SB-C18, RRHT (P/N: 827975-902) 2.1 x 50 SB-C18, RRHT (P/N: 827700-902) If customers want to cover a wide range of applications – for instance highest resolution standard bore LC/UV (e.g. 4.6x150mm RRHT) and highest speed narrow bore LC/MS (e.g. 2.1x50mm RRHT) – the system configuration may need to be changed. Depending on the degree of change such a modification takes between 5 and 20 minutes maximum. However, the standard delay volume configuration with 600-800ul pump delay volume, 0.17mm ID connecting capillaries, standard 3ul TCC SL heater and 5ul or 13ul flow cell allows to run both RRLC and HPLC on the same configuration – without changing anything!

Scatter Plot

Delay Volume (ml)0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2

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90

100

Gradient Behaviour and Method Transfer – Emulating any (U)HPLC System

Page 75

UV Signal

Method Transfer

Same RT of all Peaks

Same Resolution

all Peak Pairs

1290 Infinity LC 1200 RRLC (damper + mixer)1290 Infinity LC (+ damper + mixer + capillary)

Effects of path length increase

Peak Dispersion

60 mm pathlength

78 µL geom. volume

-

Loss of resolution-

Loss of signal height

10 mm pathlength

13 µL geom. volume

Conventional flow cells:

60 mm pathlength

4 µL σV dispersion volume

Max-Light High Sensitivity cell:

Optofluidic waveguides (total internal reflection)

Critical Comparison of TP 1.7um and SP 2.7um Particle Columns under Optimized Ultra-high Pressure Conditions. Xiaoli Wang et al, AstraZeneca, High-Speed LC Symposium, Pittcon 2009

Theory Experiment

Wed, LH5, 11:30am

78

Agilent 1290 Infinity LC Power Range and 2.7um SP Columns

bar

ml/min

2.1x50, T=80C

2.1x50, T=40C


50 x 4.6 mm100 x 4.6 mm150 x 4.6 mm250 x 4.6 mm150 x 3 mm250 x 3 mm150 x 2.1

H2 O/ACN, 40 °C

Impurity Analysis Tramadol 1200 Quat-DAD versus 1260 Quat-DAD

min1 1.5 2 2.5 3 3.5

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DAD1 A, Sig=270,8 Ref=400,100 (1260 DATA ...COLU\1200QUAT_TRAMADOL 2010-04-26 10-34-21\1200QUAT_TRAM08.D) DAD1 A, Sig=270,8 Ref=400,100 (1260 DATA ...60 NOISE 12APRIL 2010-04-26 13-52-43\1260 TRAMADOLE2_10MM5.D) DAD1 A, Sig=270,8 Ref=400,100 (1260 DATA ...60 NOISE 12APRIL 2010-04-26 12-40-03\1260 TRAMADOLE_60MM09.D)

1260 with 60mm cell

1200 with 10mm cell

1260 with 10mm cell

Imp D

Imp AImp C Imp B

Tramadol

Impurities:0.02 -0.03%Column: 50x4.6mm Zorbax

RRHT SB-C18, 1.8µm, 600bar F = 1.2mL/min

Presenter

Presentation Notes

Sample : Tramadol 4.2mg/1ml Water/ACN= 50/50 Column: 50x4.6mm Zorbax RRHT SB-C18, 1.8µm, 600bar Mobil Phase:A = Water 0.05%TFA B = Acetonitrile +0,045%TFA Step gradient:at 0 min 17% B at 2.8min 45%B Flow rate:1.2 mL/min Stop time: 3 min Post time:1min Injection volume 1µl Column temp.: 30 °C DAD: 273/4nm Ref 380/100, slit 4 Flow cell:10 mm and 60mm path length cell Peakwidth:> 0.012min (20 Hz)

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