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Jeanine Pippitt, Americas Regional Marketing Manager - HPLC

Separate Yourself from the Status Quo

Historical Perspective on UHPLC

HPLC Separation of more compounds Easier data acquisition Long runs Can’t always separate peak of interest

UHPLC Built from existing technologies No tradeoff between speed, resolution, & sensitivity Challenging to maintain and use Questionable robustness Not designed for serviceability

Completely new, differentiated UHPLC

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Every scientific breakthrough starts with a challenge.

Separate Yourself from the Status Quo

That challenge should be your science, not your instrument.

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We Questioned the Status Quo

Achieving better performance should not have to come from expending more effort.

Enables you to find better answers, more answers, and get them faster.

Easier chromatography that let’s you do what you couldn’t do before.

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• Completely new design • Leveraged 30 years of experience • Pushed beyond limitations of available

technology • Boosted robustness • New column technology • Built around the user • Leveraged our gold standard CDS software • Boosted serviceability

New Thermo Scientific Vanquish UHPLC Platform

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Confidently Control the Separation

2 Thermostatting Modes 5 °C – 120 °C Temperature Range Active Preheating

Better Separations Easy Method Transfer

Flexible Method Development

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Detect with Incredible Sensitivity

Linear up to 3000 mAU Noise levels down to ±3 µAU Lowest dispersion with Thermo Scientific™ LightPipe™ technology

See more compounds No distraction due to low

baseline noise

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Gain More Power

1500 bar (22,000 psi) of pump pressure Flow rate up to 5 mL/min 2 × 3 solvent channels technology

No tradeoff between speed and resolution

No limit to flow rate at higher pressures

Up to 9 solvent combinations to help with method transfer

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Obtain Accurate Results

Up to 23 well plates for up to 8832 samples Unsurpassed sample dosage New, patented sample pre-compression technology

Unsurpassed retention time precision and accuracy

Increased sample capacity and integrity

Low injection dispersion

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Innovation in Every Aspect of Vanquish

Integrated Modularity

Drawer module concept

Fluidics designed around the column

snap-in system assembly

integrated drainage

system talks in colors

Base drawer

SmartFlow™

ATEC

supreme accessibility of main parts Optimized seal design

Unrivaled retention time precision

Intelligent sample pre-compression

Highest sample capacity

integrated barcode reading

air-to-air cooling

Forced and still air thermostatting LightPipe™

fluorescence field

Super-wide linear range

Longlife valve

Tool-free Viper fluidics

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Using the Power of High Pressure

1500 bar

Maximize Speed Maximize Resolution

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Integrated Modularity

Sampler pre-compression

Retention time precision, Injection dispersion

4,250 4,300 4,350 4,400 4,45087250 min

4,250 4,300 4,350 4,400 4,45087250 min

Retention time precision

Detector data collection rate

• …is much more than the design of the boxes. • …combines the flexibility chromatographers need with the extra performance

from modules that are working together as one.

Ultra low pump ripple

Lower noise Improved s/n

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Retention Time Precision with UHPLC

Influence of pump-autosampler interaction

Retention Time Precision Improves on Avergage by a Factor of 6

RSD [%] 0.04 0.026 0.05 0.096 0.035 0.033 0.033 0.039 0.044 0.04

Peak 1 2 3 4 5 6 7 8 9 10 RSD

[%] 0.008 0.009 0.01 0.007 0.008 0.007 0.006 0.005 0.006 0.009

0.00 1.00 2.00 3.00 4.00 5.00 6.00

200

400

700

mA

U

min

1

2 3

4 5

6

7 8 9

10

A

0.00 1.00 2.00 3.00 4.00 5.00 6.00

200

400

700

mA

U

min

1

2 3

4 5

6

7 8 9

10

B

without pre-compression

with pre-compression

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Exceptional Sensitivity

• Impurities as low as 0.008% of the active ingredient (API)

• Exceptional linearity for API and impurities quantitation

• Very fast using 800 µL/min with 890 bar maximum pressure

• Easy method transfer from Thermo Scientific™ Dionex™ UltiMate™ 3000 XRS systems

Nevirapine & Impurities on a Thermo Scientific™ Syncronis™ C18 Column, 1.7 µm, 2.1 × 100 mm

0 60 72 -500

0

3,500

R² = 0.9998

0

1000

2000

3000

0 0.4 0.8Nevirapine [mg/mL]

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Nevirapine

18 -2

0

5

mAU

seconds

1

2

3 4

5 6 7 8

mA

U

seconds

R2 = 0.999

72

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High Throughput

• Now double as fast than before with the same flow: 800 µL/min

• Impurities still better than 0.05% of the active ingredient

• Easy method speed-up by shortening the column

Nevirapine & Impurities on a Syncronis C18 Column, 1.7 µm, 2.1 mm × 50 mm

0 20 30 -200

0

1,600

10 seconds

500

1000

1

2

3 4

0 10 20

0

10

1

2

3 4 5 6

-2

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Outstanding RT Reproducibility

Generic Exploratory Method for Traditional Chinese Medicine Columns: Thermo Scientific™ Acclaim™

RSLC C18, 2.1 × 250 mm and 2.1 × 150 mm coupled in line

Mobile A–Water Phase: B–Acetonitrile

Flow rate: 670 µL

Pressure: 1350 bar (max.)

Temp: 45 ºC (still air mode)

Injection: 5 µL

Detection: VDAD, 10 mm LightPipe data collection rate 100 Hz, 0.025 s response time, slit 4nm

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Outstanding Area Reproducibility

Sartans & Diuretics on an Accucore C18 Column, 2.6 µm, 2.1×100 mm

Column: Accucore C18, 2.6 µm, 2.1 ×100 mm Mobile Phase: A–Water, 0.1% formic acid B–Acetonitrile, 0.1% formic acid Gradient: 0-0.14 min 5%-60% B, 0.14-0.30 min 60%-90% B, 0.30-0.75 min 90% B, 0.75-0.76 min 90%-15% B, 0.76 min-1.30 min 15% B Flow rate: 1.50 mL/min Pressure: 640 bar (max.) Temperature: 60 ºC (still air mode) Injection: 3 µL Detection: VDAD, 10 mm LightPipe,

272 nm, 100 Hz data collection rate, 0.05 s response time

Analytes: Dissolved in 7% methanol 1. Hydrochlorothiazide 2. Chlorthalidone 3. Amlodipine 4. Telmisartan 5. Azilsartan 6. Valsartan 0 10 20 30 40

-50

0

100

200

300

350

1

2 3

4

5

6

sec

mA

U

Overlay of 10 Injections 0.019 % RSD

0.072 % RSD

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Unmatched Performance

Generic Routine Method for Herbal Extracts Gingko Extract on an Accucore Vanquish C18 Column,

1.5 µm × 2.1 mm × 100 mm solid core

• Very similar peak pattern with 5 times shorter column

• Excellent method for QC and routine sample

• 4 times faster than exploratory method (75 min on 500 mm column vs. 18 min on 100 mm Accucore Vanquish Column)

• Higher and sharper peaks (4 sec vs. 12 sec)

• Easy method speed-up using outstanding flow and pressure capabilities (400 µL/min at max. 850 bar)

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 -200

500

1,000

1,500

2,000

2,500

min Flow: 0.400 ml/min ACN 0.1% FA: 5.0 %

35.0

85.0

5.0

Separation of Gingko extracts on coupled Acclaim Columns (500 mm length) 700 Separation of Gingko extracts on Accucore Vanquish Column (100 mm length)

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Thermo Scientific™ Acclaim™ RSLC C18, 1.5 µm, 2.1 x 100 mm

0.01 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 4.99-6,11525,00050,00075,000100,000125,000150,000175,000200,000225,000250,000275,000300,000325,000350,000375,000400,000410,538

min

counts

Overlaid selected reaction monitoring chromatograms showing detection of 36 antibiotics within a 5 minute detection window, binary Vanquish and TSQ Vantage

Integration with MS for Targeted Screening of 36 Antibiotics

Time (min) %B

0 10

4.375 90

5.000 90

5.125 10

8.750 10

Mobile phase A: 0.1% formic acid / Water Mobile phase B: 0.1% formic acid / MeOH Flow rate: 400 µL/min Column temp.: 40 °C, active pre-heating Injection volume: 2 µL Table 1. LC gradient conditions

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Generic Methods for MS

• Peak shape and sharpness allow better identification of structurally very similar metabolites

• Generic method power allows reducing method development

• Retention time reproducibility is extremely important for this work

Full Scan Analysis of Omeprazole and Metabolites in Human Urine on Vanquish UHPLC and Thermo Scientific™ Q Exactive™ HF Mass Spectrometer using Accucore Vanquish C18 Column,

1.5 µm × 2.1 mm × 100 mm

Real Pharma Application – Stribild Combination Drug Assay

Emtricitabine pKa 2.65 211 nm, 283 nm

Tenofovir disoproxil fumarate (pro-drug) pKa 3.75 260 nm, 210 nm

Elvitegravir pKa 6.6 260 nm, 219 nm

Cobicistat pKa 1.8, pKa 2.5, pKa 6.4 195 nm, 215 nm

Stribild • A single tablet, multi active ingredients formulation

for the therapy of HIV

• Value for the patient: One pill a day

• Challenge for customer: Simultaneous determination of 4 API and their impurities

Value of the Vanquish System • Resolution to separate the API and their impurities

• Sensitivity to quantify

• Throughput with only 7 minute analysis

Real Pharma Application – Stribild Combination Drug Assay

• All actives! • Impurities • No comparable separation publicly

documented

• Outstanding linearity • Simplifies calibrations and boosts result

quality Accucore Vanquish Prototype, C18 1.5 µm x 2.1 mm x 100 mm, 0.5 mL/min, max 1200 bar, Water:MeOH (0.1%FA) gradient, T = 50 °C

• HPLC • HPLC coupled with Mass detector • UHPLC coupled with UV detector

• UHPLC coupled with Mass detector • UHPLC coupled with Mass detector &

UV detector

• HPLC grade for routine HPLC analysis

• Optima grade for routine HPLC analysis which require higher quality level

• Optima UHPLC/MS grade for UHPLC-MS which require lowest metal impurities and low baseline noise in MS detector resulting in higher signal to noise ratios

• Optima LC/MS grade for LC/MS or UHPLC-UV which require lower metal impurities

HPLC system UHPLC/MS system LC/MS system

High Quality Level

LC Instrumentation Driving Solvent Needs

Specification Comparison: Optima UHPLC-MS vs. UV

Key Take Aways: 1. Optima LCMS is scanned over the full UV wavelength, ideal for dual detection. 2. UHPLC-MS Gradient Suitability is tested without additives to ensure the highest purity. 3. Fisher Chemical is the ONLY manufacturer to ensure low baseline noise in an MS detector on every lot so that UHPLC-MS customers obtain optimum peak profiles with higher signal to noise ratios. 4. UHPLC-MS has Lower metallic impurities via borosilicate bottles packaging.

Better integration with the world’s best mass spectrometers

Maximum speed and resolution with new Thermo Scientific™ Vanquish™ LC columns

More throughput capacity by new rack loader

Revolutionary ease-of-use experience by new Thermo Scientific™ Dionex™ Chromeleon™ Chromatography Data System (CDS) features and workflows

Thermo Scientific™ Vanquish™ System

Complete Analytical Portfolio

Thank You

For more information please visit www.thermoscientific.com/separateyourself

Thank you

Now, please join me in the Vanquish section of our booth and I will address any further comments and questions.