Agilent HPLC Hardware and solution · Stepwise Upgrade - Detector Agilent 1260 Infinity Series...

Agilent HPLC Hardware and solution

January 2015

1

Xavier Maes

Product Specialist HPLC

1290Infinity II LC

LSPD Portfolio Overview

1260

Binary

1220

Gradient

1220

Isocr.

SFC1260 SFC, SFC/MS, SFC/FID

1260 Hybrid SFC/UHPLC-MS

Bio-inert LC1260 Bio-inert LC + FC

1260 Multi-Detector Bio-SEC

1290 Infinity II LC

1260

Quat.

1260

Isocr.

Dissolution708/709, 400-DS, BIO-DIS,

850-DS, UV-Dissolution, MQ

App 1, 2, 3, 5, 6, 7

High-end UHPLC

Core UHPLC

HPLC-Chip/MSUHC-NCE,PhosphoPeptide

mAB-Glyco, HiP Protein ID

Lowflow LC1260 Cap/2D-LC

1260 Nano

GPC-SEC1260 GPC, 1260 MDS,

GPC 50, GPC 220

Electrophoresis7100 CE, 2100 Bioanalyzer,

2200TapeStation,3100Offgel

Preparative LC1260 uS, 1260 AS, 1260 PS

218, SD1, A2Prep

HDR-DAD

High-Speed Flexible

Workflow Automation Solutions I2I Method Transfer / ISET, 2D-LC, Method Dev., Multi-method/Walk-up, HT LC/MS, ACR,

Online-SPE, Online Sample Prep, Online Sampling, Impurity Analyzer, A2Prep, Mobile LC…

~ 100 LC Modules

~ 30 Valves

~ 70 Accessories/Kits

~ 30 SW

Agilent LC Division

January 2015

3

Since 1970, the complete

responsibility for all HP and

Agilent LC products is located in

Waldbronn, Germany:

• Product Management

• Research & Development

• Production & Order Fullfilment

• Applications Development

• Marketing Communication

Full module compatibility

• Mix 1220, 1260, 1290 Infinity LC and 1290

Infinity II 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.

1200 Infinity Series – Agilent Value PromiseInfinitely Higher Investment Protection

January 2015

4

0

200

400

600

800

1000

1200

0 1 2 3 4 5

bar

mL/min

Confidential

Agilent 1200 Infinity & Infinity II Series Pumps- Power Range

1300

600

2 5

1220 / 1260 Infinity LC – HPLC Standard

1220 / 1260 Infinity VL

1290 Infinity LC

1290 Infinity II LC

up to 10mL/min @ 200 bar

The Agilent 1220 Infinity LC

January 2015

6

Agilent 1220 Infinity LCProduct information

600 bar for UHPLC Capabilities

Isocratic or Gradient Pump:• flow range: 0.2 - 10 mL/min

• low pressure mixing

• integrated degasser

• flow precision 0.07 % RSD

Manual Injector or Autosampler:• 600 bar

• injection range: 0.1-100 µL

• 100 x 2 mL vials or 30 x 6 mL vials

• injection cycle time 50 s

• carryover: < 0.05%

Column Oven:• up to 80°C

• one column, max. 250 mm

VWD or DAD:• 80 Hz data rate

• wavelength range: 190 - 600 nm (VWD)

up to 950 nm (DAD)

• 2.5 µAU (7µAU for DAD) noise at 230 nm

January 2015

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Integrated, yet flexible…

January 2015

8

Different configurations for different needs available!

Many upgrade paths available!

G4286B G4288B G4290B G4294B

Expandability

January 2015

9

External tray for walk-up:

Together with Agilent‘s Walkup 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!

From HPLC to UHPLC on the 1220 Infinity LCVarying Particle Size and Column Length

3/6/2016

1220 Infinity LC Portfolio

10

Column: 3 x 150 mm, 5 µm

Tg = 9 min

Resolution peak 6 = 1.23

Run time = 7.5 min

Flow rate: 1.2 mL/min at 40°C

Column: 3 x 50 mm, 1.8 µm

Tg = 3 min

Resolution peak 6 = 1.09

Run time = 3.2 min

min1 2 3 4 5 6 7

mAU

0

20

60

80

100

120

140

40

Fro

m H

PL

C t

o U

HP

LC

2.4 mL/min at 510bar

3x50 mm, 1.8µm, column at 50°C

Resolution for peak 6 = 2.53

From HPLC to UHPLC on the 1220 Infinity LCVarying Flow Rate, adapting gradient time and column temperature

3/6/2016


11

min0 0.5 1 1.5 2 2.5 3

mAU

0

10

20

30

40

50

60

70

min0 0.5 1 1.5 2 2.5 3

mAU

0

10

20

30

40

50

60

1.2 mL/min at 315bar

3x50 mm, 1.8µm column at 40°C

Resolution for peak 6 = 1.09

Fro

m H

PL

C t

o U

HP

LC

UHPLC with 1220 Infinity LC and Poroshell columns

3/6/2016


12

1220 Infinity LC1260 components in compact housing

Easy Method Transfer

Technical Overview:

„Seamless method transfer from the Agilent 1200 Series to the Agilent 1220 and 1260 Infinity LC systems”

January 2015

13

1220

1260

1100

1200

1200 RRLC

min0 1 2 3 4

mAU

0

200

400

600

800

1000

1200

1400

1200 Series LC

1220 Infinity LC

1260 Infinity LC

RT shift <5%

//nwadmc01/field_info/1_Application_Finder/2_Technical_Overviews/5990-6206EN.pdf

The Agilent 1260 Infinity LC

January 2015

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1260 Infinity LC - Most flexible LCStandard analytical LC modules

30+ different 1260 Infinity LC modules to chose from and build up YOUR specific solution

March 6, 2016

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1260 Infinity Quaternary Pump

1260 Infinity Binary Pump

1260 Infinity Autosampler 1260 Infinity Column Compartment 1260 Infinity Diode Array

Detector VL +

1260 Infinity RI Detector

1260 Infinity FLD Spectra

1260 Infinity Variable

Wavelength Detector

1260 Infinity ELSD Detector

1260 Infinity Multisampler

1260 Infinity Manual Injector

Valve options

UHPLC – Advantages & Benefits

Real UHPLC separation

performance (600 bar) benefitting

from the usage of sub-two-micron

(STM) column technology:

• Higher resolution, efficiency and

sensitivity

• Faster analysis - shorter run times –

faster turnaround times

• State-of-the-art, future-proof

instrument being more competitive

• Reduced column diameter which

helps to save solvent

March 6, 2016

16

4.6 x 150, 1.8 µm

Rs = 1.80 (+ 57%)

S/N = 44

7 Impurities

All 7 Baseline

separated!

4.6 x 150, 5 µm

Rs = 1.15

S/N = 42

4 Impurities

2 Not baseline

separated!

4.6 x 150, 3.5 µm

Rs = 1.37

S/N = 50

7 Impurities

6 Not Baseline

separated!

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)

Isocratic Impurity Method

Column: 4.6 x 150 mm, 5 µm

The value of UHPLC in Impurity ProfilingNumber of theoretical plates – Isocratic analyses

March 6, 2016

17

1100/1200

Series HPLC

Quaternary System

1220/1260 Infinity

LC

Quaternary System

1200 Series

RRLC – Std.

1260

Infinity LC

Binary System

Max Flow Rate 10 mL/min 10 mL/min 5 mL/min 5 mL/min

Delay Volume 800-1100 µL 600-900 µL 600-800 µL*600-800 ( up to 900

with VL) µL*

Capillary ID 0.17 mm 0.17 mm 0.17 mm 0.17 mm

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

Pressure Range 400 bar 600 bar 600 bar 600 bar

Injection Principle Variable Loop Variable Loop Variable Loop Variable Loop

Inj.Vol. – Std/Ext. 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL 100 / 1500 µL

Area RSD <0.25 % <0.25 % <0.25 % <0.15 %

Max Column Length 300 mm 300 mm 300 mm 300 mm* Smaller delay

volumes possible

Agilent 1260 Infinity LC (full method transferability)I-to-I Method Transferability by Design

Almost

unchanged

critical

specifications

March 2015

Agilent Confidential

18

100% Method TransferabilitySwitching from 1100 Series to 1220/1260 Infinity LC

100% HPLC compatibility: On one system you can run your established and

validated HPLC methods – unchanged – as well as push UHPLC to new levels of

performance.

March 6, 2016

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The Agilent Value PromiseHigh Investment Protection Offering Stepwise Upgrade

Full module compatibility

• Mix 1220, 1260, 1290 Infinity LC, 1290

Infinity II with 1100 and 1200 Series – and

future systems !

Stepwise upgrade – now and in future:

• Exchange Detector –

for higher sensitivity and detection speed.

• Exchange Column Compartment –

to upgrade to a fully automated multi-method

or method development solution supporting

up to 8 columns and 24 solvents

• Exchange Pump and Autosampler – for

ultra-high resolution LC on latest 1.8 µm

column technology and lowest carry-over

March 6, 2016

20

Stepwise Upgrade - Detector

Agilent 1260 Infinity Series

Diode Array Detector

Benefits

High sensitivity (10x with high sensitivity

6 cm path length cell) but low cell volume.

• Better S/N, lower LOD and LOQ.

Reduced refractive index and thermal

effects.

• Flatter baseline, better and easier

integration results.

No special care instructions for flow cell

required.

Cartridge design for convenient handling.

March 6, 2016

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History of Sensitivity Gain (DAD)- The last 30 years

Detector Model (Intro Year)

Sensitivity

Gain

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

1.4 1.8 2.0 1.7

10

0.0

2.0

4.0

6.0

8.0

10.0

12.0

1040 to 1090 1040 to 1050 1040 to 1100 1040 to 1200 1040 to 1290/1260

Agilent 1260 Infinity

Diode Array Detector

March 6, 2016

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Effects of path length increase

Peak Dispersion

60 mm pathlength

large geom. cell volume (78 µL)

lower light transmission

- Loss of resolution

- Loss of signal height

10 mm pathlength

small geom. cell volume (13 µL)

high light transmission

Conventional flow cells:

60 mm pathlength

4 µL σV dispersion volume

Max-Light High Sensitivity cell:

Optofluidic waveguides

(total internal reflection)

March 6, 2016

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1260/1290 Diode Array Detector Technical principle

Deuterium lamp

Max-Light Cartridge Cell

1290 DAD (G4212A)

Programmable slit

1024 element

diode-array

Grating

Mirror

Check DAD video on the website

1260 DAD (G4212B)

1290 DAD FS (G7117A)

NEW Fixed slit (4 nm)

March 6, 2016

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http://www.chem.agilent.com/en-US/Products/Instruments/lc/detectors/1290infinitydiodearraydetector/Pages/1290dad_video.aspx

11.6 x Higher Sensitivity1290/1260 Infinity DAD compare to 1200 Series VWD A/B

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

1290 DAD

60 mm

1200 VWD

10mm

Height [mAU] 28.87579 4.79998

Noise [mAU] 0.009806 0.01894

Signal/ noise 2944 253

Factor +11.6

1290/1260 Infinity DAD

1200 Series VWD A/B

March 6, 2016

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HDR-DAD

• 30x wider linear UV range

• Quantification of widely different concentration levels in one

single run

1200 Infinity HDR-DAD

With HDR

>30X Linearity

March 6, 2016

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History of DAD Linearity Gain The last 30 years

Detector Model (Intro Year)

1.4 1.7 2.0 1.42.9

> 30

0.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

45.0

1040 to 1090 1090 to 1050 1050 to 1100 1110 to 1200 1200 to 1290/60 1200 to HDR

Lin

ea

rity

Ga

in

(1988)(1986) (1995) (2006) (2010) (2011)

1200 Infinity HDR-DAD

With HDR

>30X Linearity

March 6, 2016

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30x Wider Linear Range with HDR-DAD3.7 mm and 60 mm and Max-Light flow cell

Conc

[µg/mL]

Detector Signal

[AU/cm]

10 x higher sensitivity ~3x higher upper limit

30x lower Limit of Detection (LOD)

30x lower Limit of Detection (LOD) for impurity analysis

- for more reliable automated peak integration

- higher area precision

60 mm

3.7 mm

March 6, 2016

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1290/1260 High Dynamic Range – HDR Increasing Linear UV Range by >30x

Technology

Cluster of two DADs

DAD 1: 60 mm Cell – for low concentrations

DAD 2: 3.7 mm Cell – for high concentrations

Output: One, combined signal, normalized to 10 mm path

length (HDR range: 0.6 x 10-6 to 6.7 AU/cm) vs for 1200

Series: 7 x 10-6 to 2 AU/cm)

Control / Usability

Like standard 1290/1260 DAD: Control,

data analysis, reporting

Investment Protection

Existing 1290/1260 DAD can be upgraded

(by 2nd DAD)

Existing 1100/1200 Systems can be upgraded

(by two 1290 or 1260 DAD)

3.7mm

60mm

Clustered DAD

DAD 1:

DAD 2:

March 6, 2016

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Fixed Dose CombinationsFrom Roche Switzerland

API 1

API 2

Fixed dose combinations:

A combination of two or more

APIs in a fixed ratio of dosesAgilent 1200 (black): API 2: 10 ml injected

Agilent 1200 (red): API 1: 100 ml injected

Infinity HDR-DAD (blue): API 1 and 2

simultaneously: 70 ml injected

March 6, 2016

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Stepwise Upgrade - Column Compartment

Benefits

More flexibility with different valve

solutions.

• Column selection for method

development or multi-method

laboratories.

• Alternating column regeneration

für higher throughput.

• Sample clean-up, pre-con-

centration, sample stripping, etc.

for automated sample preparation.

Investment protection with generic valve

drive/valve head concept.

March 6, 2016

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Agilent QuickChange Valve HeadsOverview

Available

• in various material

• for variable pressure and flow

ranges

• In addition, tailor made valves

are available on request!

March 6, 2016

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Higher Sample Throughput

Optimizing analysis times through automated

column regeneration can increase sample

throughput significantly – up to two-fold!

The determination of trace components is

always a challenge in the analysis of drug

metabolites, environmental samples, or foods

and beverages. Enrichment of the analyte prior

to analysis is often required to achieve highest

sensitivity.

Sample cleanup is essential for samples with

complex matrices, such as biological fluids, food

extracts or wastewater. Before injection into an

LC or LC/MS system, the analytes of interest

must be separated from the sample matrix.

March 6, 2016

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Faster Method ScoutingTo develop an appropriate LC method and determine

the optimum separation conditions – for example, for

chiral screening – samples must be analyzed on

several stationary phases. This process can be

accelerated significantly.

The 12-position/13-port valve can be used as a

solvent selector for analytical flow rates. It provides

access to up to 12 different mobile phases. Combined

with a column selection valve, this solvent selection

valve offers utmost flexibility for developing new

methods.

The design of the 2-position/6-port valve allows you to

switch between two flow paths. Hence it can be used

to direct the column effluent to another detector –

simply select the detector that is best suited for your

application.

March 6, 2016

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Stepwise Upgrade - Pump and Autosampler

Benefits

• Run legacy methods from 1100 or

1200 Series instrument and get

identical results without method

changes.

• Usage of sub-2-micron particle

columns for better resolution, higher

speed and better sensitivity.

March 6, 2016

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1260 Infinity MultisamplerExpanding capacity and flexibility and speed

March 6, 2016

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Key Features Benefit

Max Pressure: 600 bar

• Highest flow rates > more speed

• Small ID columns, Longer columns > more

resolution

Dual-needle option• Fastest cylce times

• Various injection volumes side by side

Injection Volume at full power range

0.1 -20ul

0.1- 40 ul

0.1-100 ul (single stroke) up to 500µL Multi-

load

• Flexibility according to customers needs

• No need to change hardware for wide volume

range injections

Lowest carryover• 30 ppm (chlorhexidine)

• 9 ppm with Multi-wash option

Flexible drawer setup• Usability

• Scalability of sample capacity and container type

Supports vials, up to 16 wellplates and

other formats as Eppendorf-Cups

• Largest flexibility for sample containers

Cooling inside of module from 4°C to

ambient

• Upgrade with compressor based cooler (slide in)

Carry-over test „Chlorhexidine“

Carry-over: 2.44 ppm

Multi-wash option

one solvent only

Carry-over: 20ppm

Std. Needle flush

one solvent only

Blue: Chlorhexidine injection

Red: Blank run after chlorhexidine injection

37

1260 Infinity AutosamplerExpanding capacity and flexibility and speed

March 6, 2016

38

Key Features Benefit

Max Pressure: 600 bar

• Highest flow rates > more speed

• Small ID columns, Longer columns > more

resolution

Needle-wash station• Outer needle wash for carryover reduction

• Carryover >40 pp,

Injection Volume at full power range

0.1 µL – 100 µL

Extended injection range option 900 µL

analytical head 400 bar

• Flexibility according to customers need

Integrated Column Compartment (ICC)

option

• Optional upgrade, user exchangeable

• Column heating (+5°C above ambient too 80°C)

• Up to two columns (30 cm lenght

• 2 Versions: ICC with 3 µL or 6 µL heater volume

Walk-Up option

• External tray Option

• Up to 5 vials

• Automated loading and disposal

Vial trays (default 2 x 66 vials 2 mL)• 2 individual trays for loading during analysis

• Optional „classic vial tray 100

Cooling inside of module from 4°C to

ambient

• Upgrade with compressor based cooler (slide in)

The New Agilent 1290 Infinity II LC

New 1290 Infinity II VWD

New 1290 Infinity II Multisampler

New 1290 Infinity II High Speed Pump

New 1290 Infinity II Flexible Pump New Accessories New 1290 Infinity II ELSD

New 1290 Infinity II Multicolumn Thermostat New 1290 Infinity II DAD

New 1290 Infinity II DAD-FS

The New Agilent 1290 Infinity II LC

Highest retention time precision for reliablepeak identification

The 1290 Infinity II Flexible Pump is the only low-

pressure mixing pump that combines the ultimate

performance of a high-pressure mixing UHPLC pump with

the flexibility of a quaternary pump. This pump delivers

superior gradient performance over the entire composition

range from 1 to 99 %.

Wide power range up to 1300 bar (2ml/min)/800 bar

(5ml/min) for optimum use of all analytical columns

independent of inner-diameter or particle technology

Flexibility for all applications

Highest retention time precision for reliablepeak identification

Isocratic analysis

The 1290 Infinity II Flexible Pump is the only low-

pressure mixing pump that combines the ultimate

performance of a high-pressure mixing UHPLC pump with

the flexibility of a quaternary pump. This pump delivers

superior gradient performance over the entire composition

range from 1 to 99 %.

Performance and Flexibility

March 6, 2016

44

The four channels of the

1290 Infinity II Flexible Pump give me

highest flexibility at a reasonable

price. Plus the performance of a high-

pressure mixing binary pump.

Technology: The Agilent 1290 Infinity II Flexible Pump

The story of the 1290 Infinity II Flexible Pump is a story about Accuracy!!

January 2015

45

3 Crucial Points to achieve ultimate quaternary pump performance (low

pressure mixing!):

1. For best performance you must know your eluents

2. New InletWeaver using proven Jet Weaver technology

3. Pump intelligence - Active Damping - with high pump

resolution (~300 pL minimum step)

The 1290 Infinity Quaternary Pumpe offers the highest accuracy and

precision over the full application range!

More automation, less manual work

March 6, 2016

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Manual mixing of different buffer

and/or modifier concentrations is

tedious and error-prone. BlendAssist

is doing that automatically and

reliably.

Technology: Blend Assist for the 1290 Infinity II Flexible

Pump

More automation, less manual work

March 6, 2016

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Influence of TFA concentration on separation of impurity

2 and on peak shape of tramadol.

Blend Assist mixes different buffer/modifier

concentrations automatically from a stock solution!

And gives the same result as for a manually mixed

solution!

Agilent innovative ISET technology allows you to use legacy

methods of virtually any LC and achieve the same results on

latest 1290 Infinity II LCs. You can retire your old systems

without retiring your precious methods.

All 1290 Infinity II pumps have ISET included.

Non-disruptive transition by ISET

ISET not only takes care for the various delay volumes in

different LC systems but also for different mixing behavior

– this is only available from Agilent and make instrument-

to-instrument method transfer easier than ever before.

Non-disruptive transition by ISET

Method Transfer to UHPLC instrumentsImpact of delay volume and mixing behavior...

Programmed

gradient step

Gradient slop/

Mixing behavior

Delay volume

1290 Infinity

Binary Pump

1260 Infinity

Binary Pump

time

response

What happens with a

programmed gradient?

Total delay volume of the system (sum

of capillaries, mixer, cells, valves..)

1200 Series

1290 Infinity

10 %

90 %

Programmed gradient

1290 Infinity LC gradient

1200 Series gradient

0 0.5 1 1.5 2 2.5 3 3.5

0

50

100

150

200

250

300

350

400

min

mAU

Injection

Same gradient conditions by moving

the gradient and by emulating the

gradient mixing behavior.

Method Transfer Possibilities Today- C) Intelligent System Emulation Technology (ISET)

Select pump

Select sampler

Intelligent System Emulation Technology ISETImplementation

Infinitely More Adaptive- Intelligent System Emulation Technology (ISET)

January 2015

53

Without ISETDifferent retention time

and resolution

With ISETEmulate other (U)HPLC system

resulting in similar retention time

and resolution.

54

Method transfer: Waters Alliance to 1290 Binary LC4.6 x 100 mm Zorbax Eclipse Plus, 5 µm, flow: 2 mL/min, 25-95% ACN-Water

Waters

Alliance

2695

345 bar

1290 Infinity

Binary LC

1200 bar

1290 Infinity

Binary LC

1200 bar

with ISET

55

Method Transfer: Shimadzu LC-20AB to 1290 Binary4.6 x 100 mm Zorbax EP C-18, 5 μm, 1.0 ml/min, 5-95% ACN-Water, 20 min, 40°C, compressibility A0.45-B1.2

Shimadzu

LC-20AB

1290 Infinity

Binary LC

1290 Infinity

Binary LC

with ISET

With the Instrument Control Frame- work (ICF) and

RapidControl drivers you can operate the 1290 Infinity II LC

under any Agilent CDS and under most 3rd party CDS.

Dealing with various CDS by ICF & RC-drivers

High capacity for different sample containers within small footprint

The 1290 Infinity II Multisampler takes a maximum load of

16 shallow well MTPs. Giving a maximum capacity of 6144

samples within the footprint of an Agilent LC stack.

Besides the maximum capacity also a high flexibility to

accept many different sample containers without re-

configuration of the system is possible.

Deep-well MTPs, large vials

Medium-well MTPs (96/384),

std. 2ml vials

Shallow-well MTPs (96/384)

High capacity, different sample containers

March 6, 2016

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Technology: Sample hotel of the 1290 Infinity II

Multisampler

Lowest carryover for highest data quality

Wash inner needle

Flow through principle washes inner needle

thoroughly and constant with eluent.

Wash outer needle

Choice of 3 external solvents to program

washing steps for the outer needle.

Needle-seat back flush

Choice of 3 external solvents to back

flush the seat.

Using the new Multiwash feature you can flush the inside

and outside surfaces of the injection needle with up to three

different solvents, as well as backflush the needle seat.

Lowest carryover for highest data quality

The 1290 Infinity II Multisampler with multiwash reduces

carryover to less than 9 ppm!


The dual-needle setup of the 1290 Infinity II Multisampler

enhances flexibility by providing two injectors in a single

instrument. You can optimize one path for large volume

injections and the other for low delay volumes.

20µl

500µl

Fast injection cycles with dual-needle injection

By fitting the 1290 Infinity II Multisampler with a second

injection needle, you can reduce cycle times to mere seconds

– virtually eliminating wait times for conventional injections and

even for large-volume loading or flushing procedures.

Capacity per benchspace

January 2015

63

System Width 52 cm 86 cm 54 cm 96 cm 60 cm 40 cm

Vial Capacity 216 486 316 648 528 432

DW Capacity 4 12 6 12 7 8

SW Capacity 4 23 6 12 21 16

CompetitorThermo

Vanquish

Thermo

Vanquish plus

rack charger

Shimadzu

Nexera 30

ACMP

Shimadzu

Nexera Sil 30

AC plus rack

changer

Waters Acquity

plus Sample

Organizer

Agilent 1290

Infinity II with

1290

Multisampler


New 1290 Infinity II Multicolumn Thermostat with space

for up to eight short columns, four 30 cm columns or

any combinations plus column selection valves allows

simplest method-switching.

Lowest dispersion for highest resolution

1290 Infinity II Diode Array Detector

• Ultra-low dispersion flow cell

• Data rates up to 240 Hz for small peak width

even in ultra-fast separations

1290 Infinity II Multicolumn Thermostat

• Quick Change heat exchangers down to 1.0 µl

• A-Line Quick-Connect fittings

• Capillary side-outlets for shorter connections to

LC/MS

1290 Infinity II Multisampler

• Optimized needle seat

min0 0.1 0.2 0.3 0.4

mAU

0

10

20

30

40

50

60

The optimized new ULD flow cell and data rates up to 240

Hz deliver uncompromised chromatographic resolution –

even for your fastest separations.


min0 0.1 0.2 0.3 0.4

mAU

0

20

40

60

80

100

Data rate = 240 Hz

Data rate = 20 Hz

More automation capabilities

Quick-Change valves for column regeneration, or sample

clean-up and enrichment. And many more applications!


Automated column and mobile phase screening with up to

8 columns within one column compartment and up to 26

solvents.


Method Scouting Wizard software for most easiest method

and sequence generation for any combinations of column,

solvent, gradient and temperature screening.

Agilent A-Line Quick Connect UHPLC column fittings for

truly dead-volume-free fluidic connections

• Tool-free connection up to 1300 bar

• Spring loaded design for easy and truly zero-dead

volume connection

• Removable and reusable for all column types


Unique detection capabilities reveal more

The 60 mm path length of the Agilent Max-Light flow cell

enables the 1290 Infinity II DAD to achieve typical noise

levels of less than ± 0.6 μAU/cm for highest detection

sensitivity and lowest detection limits.

1290 Infinity II DAD with

High Sensitivity cell

Conventional detector

Unique detection capabilities reveal more

For your samples with widely different concentration levels,

the 1290 Infinity II High Dynamic Range DAD solution

offers a 30x wider linear range and up to 30x higher sensitivity,

allowing you to detect and quantify all components within a

single run.

Two wavelengths, less injections

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75

The 1290 Infinity II VWD I can

measure at two wavelengths. I can

now use a VWD for samples that had

required a DAD so far.

Technology: 1290 Infinity II Variable Wavelength Detector

Wider dynamic range, less injections

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76

With the 1290 Infinity II HDR-ELSD I

can reliably quantify minor and major

compounds in my sample in a single

run, even if they do not have a

chromophore.

Technology: 1290 Infinity II High Dynamic Range ELSD

Select your separation performance – match system power with sample

complexity at the flip of a switch between single-dimension UHPLC and

the ultimate chromatographic power of 2D-LC.


General thoughts in separation science

78

„Resolving power is what it is all

about in analytical separation

science.“

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Bringing in a 2nd Dimension - how?

79

Pump Autosampler Column Detector

Pump

Detector

Co

lum

n

Valve

(„Sampler“)

1D-LC System

2D-LC System

+

1D-LC System

„Just like 1D-LC with a chemically selective detector“

March 6, 2016

2D-LC – Heart-cutting vs Comprehensive 2D-LC

Heart-cutting 2D-LC (LC-LC):

LC1

LC2

Parts of the 1D effluent are injected onto

2D system

Long 2D gradients possible good

data quality

Limited 2D information

80

March 6, 2016


Comprehensive 2D-LC (LCxLC):

LC1

LC2

1st peak from

1st dimension

2nd peak from

1st dimension

3rd peak from

1st dimension

The whole 1D effluent is injected onto

2D system

(Ultra)Short 2D gradients necessary

Good data quality with fast pumps &

detector

Full („comprehensive“) 2D information!

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March 6, 2016


Comprehensive 2D-LC (LCxLC):

LC1

LC2

1st peak from

1st dimension

2nd peak from

1st dimension

3rd peak from

1st dimension

LC1

LC2

min

sec

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March 6, 2016

Bringing in a 2nd Dimension - how?The unique 2D-LC valve

83

March 6, 2016

2D-LC MULTIPLE HEART CUTTING

A new tool closing closing the gap between comprehensive and

heart-cutting 2D-LC

March 6, 2016

Confidentiality Label

84

1290 Infinity 2D-LC SolutionMultiple heart-cutting

Heart-cutting Data Viewer

85

March 6, 2016

20min

4.5min

Peaks lost for extra 2nd dimension analysis

1D

2D 4.5min

1290 Infinity II 2D-LC Solution Multiple heart-cutting

86

March 6, 2016

20min

1D

2D

4.5min4.5min 4.5min 4.5min 4.5min

Unmatched multiple heart-cutting 2D-LC Usability

Pre-aligned loop-valve kits, just add

to the existing 2D-LC system

Online status monitoring

Smart Valve-Loop Setup with 12 loops

2D-LC valve + two 6/14 valves

87

March 6, 2016

1290 Infinity II 2D-LC Solution – ScalabilityMultiple heart-cutting

88

March 6, 2016

20min

4.5min

4.5min

4.5min

4.5min

4.5min

#2: 7.41 min#2: 7.41 min

#3: 9.80 min#3: 9.80 min#3: 9.80 min

#6: 12.58 min

#3: 14.39 min

#4: 12.58 min

#5: 14.52 min#5: 14.52 min

#4: 12.58 min

#5: 14.52 min#5: 14.58 min

Most intuitive software to set

up and edit methods within

seconds:

• 1st dimension gradient

• 2nd dimension gradient

• Gradient shift

• Time-segments

• Method parameter

• Method set-up calculator

• Reference chromatogram

overlay for heart-cutting

• Time-based or peak-based

mode

2D-LC Acquisition SoftwareOne easy-to-use software for all operation modes

89

March 6, 2016

New multi heart-cut data viewer for smartest heart-cut data evaluation and

automatic reporting

Data Analysis Software

90

March 6, 2016

1290 Infinity 2D-LC Solution – ScalabilityThe right solution from medium to highly complex samples

91

Heart-cutting 2D-LC

Sa

mp

le c

om

ple

xity

Multiple heart-cutting 2D-LC

Comprehensive 2D-LC

Comprehensive 2D-LC/(IMS)-MS/MS

API impurity

profiling

Complex

formulations

Bio-

pharmaceuticals

Natural products,

biological

samples....

March 6, 2016

Achiral-chiral 2D-LC of Pharmaceutical SubstancesIbuprofen (racemic)

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1st Dim:

C18

2nd Dim:

Chiral column

Method Development for Impurity AnalysisPotential Co-elutions in Onedimensional Analysis

93

Main compound

ImpuritiesImpurities

• Impurities hidden underneath the main compound peak?

• Coelutions of impurities?

Method Development for Impurity AnalysisTime-based Heart-cutting of Main Compound and Impurities

94

Method Development for Impurity AnalysisHeart-cutting 2D-LC Reveals Co-elutions of Impurities

95

Method Development for Impurity AnalysisMultiple Cuts Across the Main Compound Peak

96

• Two impurities

completely hidden

under main

compound peak

• Partial coelution of

a third impurity

with the main

compound

January 2015

97

Agilent HPLC Hardware and solution · Stepwise Upgrade - Detector Agilent 1260 Infinity Series...

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Transcript of Agilent HPLC Hardware and solution · Stepwise Upgrade - Detector Agilent 1260 Infinity Series...