920-lc analytical hplc 940-lc analytical to prep hplc ihplc

920-LC Analytical HPLC940-LC Analytical to Prep HPLC

Varian 900-LC Series iHPLC

Agenda

• Introduction• Introduction to the 9X0-LC series HPLC

• Total market size by technique and market segment

• An overview of the 920-LC Analytical HPLC• Features and benefits

• Positioning, competition and target markets

• Applications

• An overview of the 940-LC Analytical to Prep and Prep HPLC• Features and benefits

• Positioning, competition and target markets

• Applications

What is the iHPLC?

• 2 models 920-LC and 940-LC

• Compact, stylish, automated analytical and semi prep HPLC

• Pre-plumbed and factory tested for the highest confidence

• Easy to order, install and use

– Single Ethernet cable connection

• Focused on improved productivity through automation

• Updated premium performance hardware

• Autosampler, Fraction Collector and Scale up Module for semi-prep

• Quaternary pump, precision Column Heater,RI, Fluor, ELSD and autosampler for analytical

• Updated Integrated Instrument Software Interface

• Eliminate repetition, modernise UI

• Consistent

• Easy to use with added functionality

• Adds to modular range

HPLC DemandSDI 2007

Service16%

Initial Systems

Aftermarket46%

Fast LC10%

Other9%

Prep14% Conventional

HPLC63%

• Overall LC is $3.2B @ 6% growth

• 23000 systems sold in 2006

• New HPLC systems sales (initial)

• $1220M

– $764M Conventional analytical

– $180M Prep

– $113M Fast LC

Current Status of Varian HPLC

• Installation/setup difficulties• No supporting materials• Perceptions• Ease of ordering• Competitive performance• Lack of competitive specifications• Ease of use• Reliability• Productivity• Validation and Compliance• Flexibility/expansibility (especially in semi-prep)• Competitive pricing• Wide product range – hardware, columns, consumables

Varian 920-LC Integrated Analytical HPLC

Varian 920-LC Silver Bullets

• Premium Analytical Pump Performance and reliability

• Superb Fast LC Performance optimised for sub 3 um columns– Improved throughput without compromising robustness

• High sensitivity/Top Line Performance detection systems across UV/PDA/RI/Fluor and ELSD – Unparalleled strength

• Unbeatable Injection performance and carryover statistics with 2008 Loopless Technology Autosampler

• Full system level computer controlled AIQ (validation)

• Updated UI for Enhanced User Experience

Varian 920-LC Analytical HPLC Goals

• Aim

• Become credible player in this $800M segment by doubling market share and revenue

• Markets

• Pharma / BioPharma

• Environmental

• Industrial

• Strategy

• Leverage Varian distribution channel to deliver competitive HPLC hardware, consumables, software and services

Varian 920-LC What Is It?

• Compact, stylish automated analytical HPLC

– Pre-plumbed and factory tested for the highest confidence

– Single cable communications

• Virtually pulse-free dual piston pump

– Does not require a pulse damper

• State-of-the-art low pressure quaternary gradient module with built-in degasser

• Autosampler with cooling option

– Synchronised injection for outstanding retention time reproducibility

– Compatible with 200 x 1.6 ml HPLC vials or 3 microplates

Varian 920-LC What Is It?

• Either dual UV-Vis or PDA detector

– Optional second detector - Fluorescence or RI

• High precision (<0.15oC) dual column heater available

– Low delay solvent pre-heater for improved heat transfer

– Optional column switching and electric injection valve

• NEW improved instrument interface on GalaxieTM software

• External CVM, ELSD or 356-LC RI can be plugged directly into the back of the unit.

920-LC for Standard HPLC to Fast HPLC

25cm x 4.6mm Pursuit XRS 5um 1.0ml/min

10cm x 2.0mm Pursuit XRS 2.8um Ultra 0.8ml/min

5cm x 2.0mm 2.4um 1.3ml/min16 x Speed Enhancement

920-LC for Standard HPLC to Fast HPLC

Varian 920-LC Fast LC PAHs

Varian 920-LC6 minute Analysis

Waters Acquity7 Minute Analysis

Varian 920-LC Pump

• Dual piston quaternary pump produces fast gradients – even at 200 l/min

• Virtually pulse-free pumping without pulse damper

• Outstanding gradient compositional accuracy give retention time precision at the <1% level

• Built-in vacuum degas maintains pump prime and enhanced injection reliability

• Accurate flow from 0.1 to 10 mL/min for 2 to 4.6 mm columns

• Overlay of 10 chromatograms

• 10% steps at 1ml/min

Varian 920-LC Pump - FAB

Feature Advantage Benefit

Dual Piston Quaternary Capability

Market leading precision and accuracy

Performance benchmark

Built in Degasser and Pulse Free Flow

Enhanced run to run reproducibilty

Dependable

Confidence in results

Optional Semi-Micro

Fast LC capability Performance with Reliability

Varian 920-LC Autosampler

• Needle-in-flow path geometry for low carry-over (<0.01%)

• Typical inj precision (< 0.3% at 10 l)

• Optimized flow paths for fast injection turn-around

• Synchronised injection for best RT reproducibility

• Programmable wash setting for low carry-over

• Injection volumes from 1ul to 50L

• Cooled option available

Varian 920-LC Washout Performance

Blank injection <0.01% carryover

125 ug/ml Caffeine

920-LC Autosampler How Does it Work?

Varian 920-LC Injection Synchronization

0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2

Varian 920-LC Autosampler - FAB

Needle in Path

Design

No need for Injection loops

No Carryover

Versatility

Performance

Synchronised injections with Pump Stroke cycle

Enhanced retention time reproducibilty

Performance

Accuracy

One Autosampler, all rack formats

No need for multiple injector options

Versatility

Simplicity

Sensitive UV-Vis Detectors

Typical Agilent VWD S/N ~ 25

Typical Varian 920-LC UV Vis S/N ~ 38

Varian 920-LC UV/Vis - PDA Detectors - FAB

Best signal to Noise Enhanced sensitivity and detection limits

Performance

Tool Free maintenance

Easy to change lamps and flow cells

Simplicity

Range of cells for ul/min to Litres/min

Fits all applications from LCMS to Benchtop Prep

Versatility and Performance

Varian 920-LC Fluorescence Detector

• Sensitive Fluorescence detector (S/N > 600 Raman H2O peak baseline method)

• Time programmable for increase sensitivity• Automatic validation with built-in Hg lamp• Low 12 l dead volume flowcell for improved

chromatography

Varian 920-LC Fluorescence Detector - FAB

Improved Sensitivity

(S/N > 600 Raman)

Lower limits of detection

Performance

Automatic calibration

( Hg lamp)

Greater accuracy of analysis

Confidence in results; Dependable

Low dead volume cell ( 12ul volume)

Sharper peaks, enhanced resolution

Performance

Varian 920-LC Refractive Index Detector

• Same as 356-LC Refractive Index Standalone

• Low Volume, flow efficient cell for Superior Peak Shape

• Stable, Long Life and High Intensity LED

• Temperature Controlled Optics for minimised baseline drift and outstanding detection limits

• Complete automation thru Galaxie Software

Varian 920-LC RI Detector - FAB

Low Volume Cell Minimal Peak Dispersion giving enhanced resolution

Performance and Accuracy

High Intensity, long life LED

Maintenance Free for consistant long term performance

Dependability, Durability and Confidence in results

Temp Controlled optics

Minimises baseline drift for lower settling times, better sensitivity

Performance and Ecconomy – saves the analyst precious time

Varian 380-LC and 385-LC ELSD

• Varian Product numbers

• 2 to 3 Times More Sensitive

• NEW Glass Nebuliser for Improved Reproducibility

• 40Hz Data Rate for Fast LC

• Onboard timetable program to enable parameter change thru a run

Varian 380-LC and 385-LC ELSD - FAB

Improved Sensitivity Lower limits of detection

Performance

NEW Glass Nebuliser

Greater run to run reproducibility

Confidence in results; Performance

40Hz Data Rate Suitable for Fast LC Performance coupled with 920-LC in Fast LC operation

Heated Column Compartment

• Temperature range from ambient + 10oC up to 65oC

– Temperature reproducibility +0.15oC

– Ensures excellent analytical reproducibility

– Uses a low delay volume pre-heater for efficient heat transfer to mobile phase

– Spacious column compartment easily accommodates 2 x 250mm columns

– Option with electrically actuated column switching valve and injection valve

Outstanding Stability

Mobile phase input Temp

Mobile phase temp inside column

Oven Temp

Reproducibility of 50 Injections

Reproducibility <0.8% over 50 injections and 12 hours

Varian 920-LC Column Heater - FAB

Low Volume Solvent Pre-Heater

Eliminates thermal column effects and enhances resolution/method transfer

Performance

Outstanding Stability and Accuracy

Provides greater method robustness

Dependable, Confidence in results

Accurate

Spacious compartment with easy access

Fast column changes, room to work

Simple and Convenient

Varian 920-LC Software Usability

• User friendly GalaxieTM software

• Simplified, modern, consistent Instrument interface

• Modular 11 pages

• 920-LC 4 pages!!

• One page real-time status and chromatogram

• Advanced integration and calibration

• New IMS capabilities

• Automated instrument validation

• Reduces downtime and operating costs

• Fusion AE

Varian 920-LC AIQ (Validation)

AIQ (Analytical Instrument Qualification)• Fully PC controlled IQ and OQ (not PQ)

• Puts the Varian 920-LC at the forefront of regulated capabilities

• Consistent, reproducible methodology• Includes pumps, autosampler, detectors, column

heater

• Chargeable option

• Fully documented

Varian 920-LC User interface - FAB

Modern, User friendly UI

( 11 pages to 4!)

Simple to use and an enjoyable experience

Simplicity coupled with Stylish Appearance

Shockwave Graphics for 2008 Software Technology

Visually pleasing with Flashing instrument status

Stylish appearance

Instrument maintenance Software

Automated instrument validation/testing

Ensures instrument Accuracy and Performance

Varian 920-LC Concept - FAB

1st Fully Integrated

iHPLC System

Unique and Interesting concept

Prestige resulting in elevated HPLC interest/prospects

Easy to Use, Install and Maintain

Creates a new level of customer satisfaction

Product Confidence and enhanced repeat business

Compact and Stylish High level of instrument capability in Unique footprint

Desirable to own, Convenient to use

Single Part Numbers Easy to quote Much simpler to sell, and easier experience

One Power, Communications cable and “On” Button

Easy to configure, clean and tidy look

Simplicity with enhanced Appearance

Pre-Plumbed and “Ready” out of the Box

Fast installation so you are quickly able to use the instrument

Convenient and Simple

Sample tray access without opening the main doors

System looks good at all times without exposing internals

Clean Appearance

Door trays and holders for columns and accessories

Provides space to tidily store the many parts used daily

Clean Appearance

Tray Storage Provides additional storage space for Software CD, tools, columns etc…

Clean Appearance and Versatility

Solvent Tray capacity

Shape of instrument provides ample space for solvents

Convenience and Safety

Ski Slopes for easy tray insertion/removal

Eliminates tray location errors

Provides Autosampler Confidence and Durability

Single Centralised Waste port

No need for multiple waste vessels positioned around the instrument

Clean Appearance and Simplicity.

Only 1 waste to empty

Leak Detector Ensures the system is “leak free” and performing to the max

Performance and Confidence

Modern Manual Injectors

Push button valve positioning with automatic return, eliminate operator error

Even Simpler to use, eliminates user error improving Confidence in results

Bottle Management Ability to fill reservoirs and empty waste

Provides a Safety mechanism for Confidence in overnight runs

Tool Tips, Help and Videos

Extensive library of Mouse tool tips and Online help

Makes the instrument Simple to use and Convenient to access

New Brochures Physical material to present to customers

Convenient and Powerful messages to explain to customer

Updated Data Sheets, Price List and App Notes

Supporting materials to leave with the customer

Provides Confidence in product range and Simple to Quote

Varian 920-LC Ordering Matrix

pump inject detector Oven Detector 2

option

Options

Modular Isocratic 210-LC 10ml Manual 325 PDA, RI, ELSD

Isocratic

00-100890-00Dual piston pump 10 ml head

Auto Dual UV/Vis

Opt FL, RI ELSD

Gradient

00-100891-00Quat, Grad, Degas Auto Dual

UV/VisOpt FL, RI ELSD opt

High Performance Gradient

00-100892-00

Quat, Grad, Degas Auto cooled Dual UV/Vis

Opt FL, RI ELSD opt

Development

00-100893-00Quat, Grad, Degas Auto PDA Std FL, RI ELSD opt

Research

00-100894-00Quat, Grad, Degas Auto cooled PDA Std FL, RI ELSD opt

Varian 920-LC Target Markets

• Strong historical position for Varian in analytical

• Varian has the leading range of analytical technologies: spectroscopy, chromatography including consumables, data and services

• Target Users

• Environmental

• Food/Beverage

• Industrial

• Generic Pharmaceutical *

• Academia and Biotech

• Same (but wider) target groups with a stronger product offering

*40% of Regulated Pharma has significant competitive barriers due to installed software base

Varian 920-LC Consumables

• Excellent range of HPLC Columns providing industry leading resolution and robustness

• Pursuit

• Pursuit XRS

• Polymeric Phases - PL

• From 2mm to 2” ID Prepacked

• Other Speciality Phases

• NEW 2.8 Ultra Column AND 2.4um Columns for Fast or High Res LC

• 2.8um Ultra is optimised for Speed

• 2.4um gives further enhancements in Resolution

2.8um Vs 2.4 um – High Res or Extra Speed

2.8um 2mm x 5cm for speed

2.4um 3mm x 5cm for RESOLUTION

Varian 920-LC Positioning

UPLC, Fast LC and the Varian 920-LC

What is Fast LC?

• Involves using LC columns packed with very small (sub 3 m diameter) particles

• Smaller particles column are more efficient giving greater resolution per unit length

• For the same separation (at the same flow rate) can use a shorter column which reduces run time for improve productivity

• As Van Deemter plot is flatter can operate at higher flow rates efficiently

But smaller particles give greater back pressure!!

Van Deemter Plot

Linear velocity u cm/sec

0.0 0.2 0.4 1.61.0 1.2 1.40.6 0.8

Fast LC Productivity

Market Size for Fast LC - 2007

2007 HPLC Demand by Product Type

Other9%

HPLC61%

Prep LC15%

Fast LC11%

Total Systems market Size 2007 = US$1.3b (SDI 2006)

Fast LC

• Fast LC market estimated at $160mill growing rapidly• Many names

• Ultrahigh Pressure LC

• Rapid Resolution LC

• High Resolution LC

• Fast LC

UPLC• Research Pharma – Drug discovery• QA/QC Pharma – Not as successful as first thought• No.1 use - separation front end for MS

Target Markets for UPLC and Fast LC

UPLC or Fast LC• Small particle porous column packing 1.7 – 3.0 m in

diameter• Increased flow rates• Increased pressure• Increased column temperatures

Advantage:• Improved resolution of peaks in complex samples• Improved sensitivity for low concentration or limited

samples• Reduced analysis times

Theory

UPLC or Fast LC• Small particle porous column packing 1.7 – 3.0u in dia.• Increased flow rates• Increased pressure• Increased column temperatures

Disadvantage:• Increased back pressure – special hardware requirements

needed, micro leaks become a problem = Poor reproducibility

• Band spreading in the column decreases – the HPLC will need to account for this

• High acquisition rates for detector collection

• Expensive

• Proprietary columns in some instances

Theory

• Price – Binary gradient LC with autosampler and detector ~

50-60% of UPLC cost

• Flexibility– Operates as standard HPLC and in Fast LC mode

• Allows all LC applications to be run

• Reliability– Reproducibility of results

Advantages of Fast LC

• Speed – Reduced analysis times = cost savings

• Flexibility– Optimum performance as front end for MS

• Performance– Improved sensitivity

Advantages of UPLC

Typical Speed Performance of a UPLC Separation

Typical Speed Performance of Fast LC

2.4 minutes

Fanta Orange Drink

Sugar Standard

2.72.62.52.42.32.22.121.91.81.71.61.51.41.31.21.110.90.80.70.60.50.40.30.20.1

RT [min]

mAU Soft drinks_19.DATA [PDA detector Absorbance Analog Channel 1 ]

Conditions

Mobile Phase: 10mm Sodium Phosphate ph6.0/ACN gradientDetection: UV DetectionTemperature: 40°CFlow-Rate: 1,2mL/minColumn: Varian Ultra 50x2,0mm

Waters Acquity• High pressure – quoted to 15,000psi but generally

run at around 10,000psi• Low dispersion• Small particle size columns• Small columns – generally <2mm in dia.• Binary gradient system – 4 solvent selection per

pump• Typical column temp of 30°C to 50°C

Current Systems on the market

Agilent 1200 RRLC• High pressure – quoted to 9,000 psi but generally run

at lower pressures• Low dispersion• Small particle size columns• Flow rate maximum of 5 ml/min• Binary gradient system – 4 solvent selection per

pump• Typical column temp of 30°C to 100°C• Can switch between Fast LC and normal LC (variable

dead volume control).

Thermo Accela• High pressure – quoted to 15,000psi only with flow

rates of 1-1000 L/min• Low dispersion• Compatible with small particle size columns• Promoted as conventional and Ultrahigh press LC

– max flow rate of 1ml/min limits conventional HPLC

• Quaternary pumping system

Others

Shimadzu• Conventional LC that shows 2.2 m particle column

applications – Fast LC approach

Jasco• UPLC system utilising 15000psi pressure capability

and column temperatures to 60°C

Hitachi• 9000psi pressure capability with 90°C temperature

performance. Variation on their L2000 series HPLC

Van Deemter Plot

Linear velocity u cm/sec

0.0 0.2 0.4 1.61.0 1.2 1.40.6 0.8

Trends in Fast LC

• Movement away from sub 2 m particles owing to– Ultrahigh back pressures & reproducibility– High temperatures & column lifetime– Data acquisition rates

• 2.4 m and 2.8 m particles allows– Improved resolution without ultrahigh back pressures– Use of modified conventional HPLC (reduced delayed

volumes)– Use of more moderate conditions e.g. temp & pressure

• Many column and instrument companies developing products– Shimadzu– Phenomenex– Varian

16 PAHs by UPLC and Fast LC

Waters Acquity UPLC

Varian 920-LC Fast LC

Mobile Phase: Acetonitrile Water GradientDetection: UV@254nmTemperature: 40°CFlow-Rate: 0,8mL/minColumn: Pursuit XRs C18 3um, 100x2mm

Soft Drinks Using 920-LC

2.4 minutes

Fanta Orange Drink

Sugar Standard

2.72.62.52.42.32.22.121.91.81.71.61.51.41.31.21.110.90.80.70.60.50.40.30.20.1

RT [min]

mAU Soft drinks_19.DATA [PDA detector Absorbance Analog Channel 1 ]

Conditions

Mobile Phase: 10mm Sodium Phosphate ph6.0/ACN gradientDetection: UV DetectionTemperature: 40°CFlow-Rate: 1,2mL/minColumn: Varian Ultra 50x2,0mm

Alkylated Phenones Using 920-LC

1.4 minutes

1.81.751.71.651.61.551.51.451.41.351.31.251.21.151.11.0510.950.90.850.80.750.70.650.60.550.50.450.40.350.30.250.20.150.10.05

RT [min]

mAU Phenones_31.DATA [PDA detector Absorbance Analog Channel 1 ]Conditions

Mobile Phase: Acetonitrile Water GradientDetection: UV DetectionTemperature: 60°CFlow-Rate: 1,6mL/minColumn: Varian Ultra 3um 50x2,0mm

Anthocyanadins Using 920-LC

2,221,81,61,41,210,80,60,40,20

RT [min]

mAU anthocyanidins mixtureHS_16_3.DATA [PDA detector Absorbance Analog Channel 2 ]

ConditionsMobile Phase:-solvent A Water phosphoric acid 0,3% -Solvent B Acetonitrile -Gradient separation from 88% A to 78% B in 2,5 minDetection: DAD 525 nmTemperature: 55°CFlow-Rate:0.9ml/minColumn:HP 3 C18 CPInjection volume : 1µl sample Delphinidin, Cyanidin, Petunidin, Pelargonidin, Peonidin, Mavidin

Fat Soluble Vitamins Using 920-LC

11109876543210

0 RT [min]

liposolubles vitamins C18 10cm_20_2.DATA [PDA detector Absorbance Analog Channel 1 ]mAU

11109876543210

0 RT [min]

liposolubles vitamins C18 10cm_20_2.DATA [Max. absorbance betw een [210,00 ; 500,00] nm]mAU

Chromatogram 260 nm

Chromatogram MaxPlot

Conditions

Mobile Phase: A Water 0;1% formic acid B CH3CN 0,1% Formic acid gradient t 0min B 80% ; t 1.5 min B80%; t 2min B 100% T 12 min B100%Detection: UV 260 nm & MaxPlotTemperature: 50°CFlow-Rate: 0,7mL/minColumn: Pursuit C18 3 µm 100 x 2,1mmInjection volume 25 µL, sample Retinol, vitamin D2,D3,E, E Ac, K1

Fast Sugars Using 920-LC

Sprite

32.92.82.72.62.52.42.32.22.121.91.81.71.61.51.41.31.21.110.90.80.70.60.50.40.30.20.10

RT [min]

µRIU Fanta_25.DATA [RI detector Channel]

Fanta Orange Drink

32.92.82.72.62.52.42.32.22.121.91.81.71.61.51.41.31.21.110.90.80.70.60.50.40.30.20.10

230220210

200190180170160

150140130120

110100908070

60504030

-20-30-40-50-60

-70-80-90

-110-120

A 1.00

Fructose

RT [min]

µRIU Sugar Mix_50uL_41.DATA [RI detector Channel]

Sugar Standard

Conditions

Mobile Phase: Acetonitrile Water 75:25Detection: RI DetectionTemperature: 40°CFlow-Rate: 2,5mL/minColumn: Polaris NH2 5um 150x4,6mm

Varian 920-LC Conclusion

• UPLC does not currently suit all applications• Restricted column choices

• Demanding H/W are chemistry requirements

• Fast LC with 2.4 m to 3 m particle columns provides flexibility

• Compatible with 920-LC

– Low delay volume kit e.g mixer, flowcell, tubing

• Wide choice of chemistries

• Price competitive (can purchase 2 Fast LC for UPLC)

Varian 920-LC Target Markets

SDI 2006 Market by Industry Split

Other, 3%

Petrochem, 3%

Polymers, 4%

Ag/Food, 6%

Pharma, 28%

Indep Test, 10%

Industrial, 10%

Biotech, 11% Gov, 11%

Academic, 14%

Varian LC Sales by Industry Type

Academic30%

Pharma22%

Industrial 15%

Food/Ag4%

Enviro9%

R & D3%

Govt9%

other 8%

Varian 920-LC Target MarketsEnvironmental

• Build-on Spectroscopy, GC-MS & LC-MS products– Superb gradient pumping system

– Sensitive Fluoro detector for PAH’s

Varian 920-LC Target MarketsIndustrial

• Traditionally strong Varian market• Leverage profile of GC, Spectroscopy and Polymer

Lab products• Large range of systems

• Isocratic system with UV, RI, PL columns, standards & Galaxie add-in ideal for low temp GC applications

• 920-LC has a wide range of detection incl:

– UV, PDA, FL, RI, ELSD

Varian 920-LC Competitors

Varian 920-LC Market Competitors

Competitors• Shimadzu• Waters• Agilent• Dionex

•The 920-LC presents a significant step forward for the Varian

Varian 920-LC Compared to the Waters Alliance HPLC

Waters Alliance

• Consists of 2695 Separations Module• Pump, autosampler and controller

• Options of– 2996 Diode Array Detector

– 2487 Dual wavelength Detector

– 2475 multi-wavelength Detector

– 2414 RI Detector

– 432 Conductivity Detector

– 2420 ELS Detector

– 2465 Electrochemical Detector

Varian 920-LC Compared to Waters Alliance

Waters Alliance Varian 920-LC

Maximum flowrate 5 ml/min 10 ml/min

Maximum pressure

Autosampler Capacity

120 HPLC vials 200 HPLC vials

Microplate Capability NA 3 plates

Maximum Column Heater Temp

60oC 65oC

Number of Detectors None Up to 2 built-in

Sample Carryover <0.1% <0.01%

Varian 920-LC compared to the Agilent 1200 Series LC

Agilent 1200 Quaternary Pump G1354A

• Dual piston, low pressure quaternary gradient pump• Electromechanical check valve

• Variable piston stroke

• Pulse damper between primary and secondary piston

• Upgrade?? of 1100 series• Requires degasser module

Agilent 1200 Autosamplers

• Option of • G1329A standard Autosampler capable of 100 HPLC

• G1367B Enhanced Autosampler compatible with either 2 microplates or 108 HPLC vials

• Standard carryover <0.1% but can be improved to <0.01% using needle wash

• Both have peltier cooled option

Agilent 1200 Detectors

• Wide range of detectors• Variable Wavelength Detector

• Multi-Wavelength Detectors

• Diode Array Detector

• Fluorescence Detector

• RI Detector

Varian 920-LC Compared to Agilent 1200

Agilent 1200 Varian 920-LC

Pulse Damper Yes Not Required

Microplate compatibility

Not Compatible Up to 3 96 well plates

Autosampler Capacity

100 HPLC vials 200 HPLC vials

Dual UV-Vis Capability

NA Yes

Carryover <0.1% <0.01%

UV-Vis range 190 – 600 D2 only 190 – 900 D2 & QI

UV detector noise + 0.75 x 10-5 Au + 0.5 x 10-5 Au

Applications of the 920-LC

Improved Analysis of 18 Polycyclic Aromatic Hydrocarbons using a Varian 920-LC Analytical

HPLC and Pursuit® 3 PAH Column

By Phuong TruongVarian Australia Pty Ltd

679 Springvale Rd Mulgrave VIC Australia

Varian 920-LC Analytical HPLC

Introduction

• PAH’s are carcinogenic contaminants produced as a by-product of combustion

– Found in drinking water, soils and sludges

• Can be analysed by GC, GC-MS• Compared to GC, HPLC offers

• Easier sample preparation

• Improved sensitivity with FLD

• Better selectivity

• PAH’s are difficult to analyse because of their structural similarity

• Results in long gradient runs of 25 to 30 minutes

Instrumentation

• Varian 920-LC Analytical HPLC (0010089300)• Quaternary gradient pump with 4 channel degasser

• 200 position autosampler with 100 L syringe

• Thermostated column compartment

• Diode Array detector

• Optional Fluorescence detector (99100xxx00)• Varian Pursuit® 3 PAH 100 x 4.6 mm column with

3m packing material (A7001100X046)

Instrument Conditions - Gradient Conditions

Time (min) Flow rate

(ml/min)

% Water % Acetonitrile

0.0 1.3 40 60

3.0 1.3 40 60

7.0 1.3 10 90

13.3 1.3 0 100

16.0 1.3 0 100

16.1 1.3 40 60

21.0 1.3 40 60

Instrument Conditions - Fluorescence Detector Conditions

Time (min) Excitation

Emission

0.0 270 323

5.6 250 375

7.1 270 410

8.5 265 380

10.5 280 420

14.7 293 498

Results

Component UV LOD

Fluoro LOD

USEPA RDL

Naphthalene 2 0.04 1.8

Acenaphthylene 5 NA 2.3

1-Methylnaphthalene

5 0.05 NR

2-Methylnaphthalene

5 0.06 NR

Acenaphthene 8 0.03 1.8

Fluorene 2 0.003 0.21

Phenanthrene 0.5 0.04 0.64

Anthracene 0.5 0.06 0.66

Fluoranthene 1 0.14 0.21

Results

Component UV LOD

Fluoro LOD

USEPA LOD

Pyrene 1 0.02 0.27

Benzo(a)anthracene 1 0.01 0.013

Chrysene 0.5 0.04 0.15

Benzo(b)fluoranthene 1 0.01 0.018

Benzo(k)fluoranthene 1 0.01 0.017

Benzo(a)pyrene 1 0.01 0.023

dibenzo(ah)anthracene

2 0.01 0.03

Benzo(ghi)perylene 2 0.01 0.076

Indenol(123-cd)pyrene 0.5 0.02 0.043

18 PAH by Varian 920-LC & Pursuit 3 PAH

16 PAH by Dionex HPLC

• Gradient separation at 1.5 ml/min with Fl Detection

17 PAH by Grace Vydac 201 TP 5 Column

• Flow rate 1.5 ml/min

Conclusion

• The Varian 920-LC and Pursuit 3 PAH column are capable of separating all 18 PAH’s in 15 minutes

• The reproducible gradient delivery and thermostated column compartment ensure excellent retention time reproducibility

• With the optional built-in fluorescence detector, detection limits exceed the requirements of USEPA method 8310

• Offers improvements in throughput compared to alternative HPLC separations (Grace Vydac 201 TP 5 column)

Analysis of Ibuprofen in Tablets According to the US Pharmacopoiea using a Varian 920-LC Analytical HPLC and Pursuit®XRs column

By Phuong TruongVarian Australia Pty Ltd

679 Springvale Rd Mulgrave VIC Australia

Varian 920-LC Analytical HPLC

Introduction

• Assays of active pharmaceutical ingredients (API) are common in the pharmaceutical industry

• Often used to quantitate the API and contaminants• Ibuprofen is a NSAI in widespread use around the world• 4-isobutylacetophenone (Ibuprofen related compound C) is a

breakdown product of Ibuprofen• It can adversely affect the central nervous system• The USP requires Ibuprofen related compound C to be

determined at < 0.1% per tablet• USP prescribe

• a 250 mm x 4.6 mm ID x 5 m L1 type (C18) column• Valerophenone internal standard

• In this work the recommended column was compared to a 100 x 4.6 x 3 m column

Instrumentation

• Varian 920-LC Analytical HPLC (0010089300)• Quaternary gradient pump with 4 channel degasser• 200 position autosampler with 100 L syringe• Thermostated column compartment• Diode Array detector

• Columns• Varian Pursuit®XRs columns feature 100Å silica, high

sample loadability, extended pH range, outstanding resolution and excellent column lifetime

– Varian Pursuit® XRs 100 x 4.6 mm x 3m column (A6011100X046)

– Varian Pursuit® XRs 250 x 4.6 mm x 5m column (A6010250X046)

Instrumental Conditions

Mobile Phase 1% chloroacetic acid in water pH 3/Acetonitrile 40:60 v/v

Flow rate 2 ml/min

Wavelength 254 nm

Temperature Ambient

Definitions – Resolution and Selectivity ()

Chromatographic Resolution and Selectivity

• Resolution R is similar to selectivity but takes into account peak width

• Baseline resolution is when R=1.5

• Capacity factor • Selectivity

])()[(2'

Standards - Pursuit®XRs 250 x 4.6 x 5m Column

• Ibuprofen and Internal Std with 250 mm x 5 m column

Standards – Pursuit®XRs 250 x 4.6 x 5m Column

• Internal Std and Ibuprofen related compound C with 250 mm x 5 m column

Results – Pursuit®XRs 250 x 4.6 x 5m Column

Resolution

(USP > 2.5)

Assymetry

(USP < 2.5)

Selectivity

Ibuprofen 1.29

I/S in Ibuprofen solution

7.3 1.00 1.29

I/S 1.06

Ibuprofen related C solution

5.1 1.01 1.18

• Ibuprofen and Internal Std with Pursuit® XRs 100 mm x 3 m column

• Internal Std and Ibuprofen related compound C with Pursuit® XRs 100 mm x 3 m column

Results – Pursuit® XRs 100 x 4.6 x 3m Column

Resolution

(USP > 2.5)

Assymetry

(USP < 2.5)

Selectivity

Ibuprofen 1.27

I/S in Ibuprofen solution

5.3 1.01 1.29

I/S 1.13

Ibuprofen related C solution

4.0 1.10 1.18

Ibuprofen Tablet

Chromatogram of Ibuprofen tablet sample spiked with I/S using a Pursuit® XRs 100 x 3m column

Spectral Confirmation using PDA Detector

Data for Ibuprofen Tablets

% of labelled Concentration of Ibuprofen

Assymetry of Ibuprofen

Assymetry of I/S

Resolution between Ibuprofen and Valerophenone I/S

101.9% 1.27 1.08 5.4

Conclusion

• The Varian 920-LC and Pursuit® XRs 3m columns significantly improve throughput of an Ibuprofen assay compared to the USP recommended columns

• The calculated concentration of Ibuprofen in the tablet complies with USP requirements of nominal + 10%

• The sensitivity of 920-LC PDA detector ensures the LOD for the Ibuprofen related C compound is 10 fold lower than USP

• The PDA detector allowed positive confirmation the unknown peak in the tablet was Ibuprofen related compound C (4-isobutylacetophenone)

Varian 920-LC All Your HPLC needs

• The 920-LC

– Is a robust high performance automated Analytical HPLC

– Fast LC compatible using 2.4/2.8 m columns

– Has a state-of-art LP quaternary gradient solvent delivery system

– A high capacity, low carry-over autosampler with leading performance specifications

– Features sensitive UV-Vis and PDA detection technology for unmatched performance

– An option of second detector fitted – Fluoro or RI

– Uses highly capable GalaxieTM software with a simpler, consistent instrument interface for real productivity

– Has automated AIQ and maintenance tools

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