simplified liquid extraction faster. easier. reliable. sle ......simplified liquid extraction n o vu...

novumsimplified liquid extraction

Faster. Easier. Reliable. SLE Simplified.

Need Ultra-Low Detection Limits? Move Over Biotage® ISOLUTE® SLE+. Meet New Novum PRO.

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PATENT PENDING





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EN

-RU

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0132

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Your Diatomaceous Earth SLE Product Could Use Some ImprovementsThe material inside your SLE product consists of diatomaceous earth (fossilized diatoms) that is extremely old and can make extractions unreliable.

© 2021 Phenomenex, Inc. All rights reserved.

All diatoms are unique, making each of your traditional SLE products unique and therefore inconsistent.

Diatomaceous earth can be found in various mines across the world. Do you know where each batch of your SLE media comes from?

1000x zoom of salt water vs. fresh water diatomaceous

earth sorbents

2

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Redefine Your SLE with Novum™ Simplified Liquid ExtractionNovum SLE (patent pending) eliminates the questions associated with diatomaceous earth. As a lab manufactured sorbent, you can expect reliable, more consistent results.

And Now Introducing Novum PRO SLEThe same reliable SLE sorbent with our new manufacturing capabilities to help you

achieve low limits of detection with clean, high-throughput extractions.

Proudly manufactured at

PhenomenexHeadquarters in

Torrance, CA USA

3


Visit www.phenomenex.com/Novum for buffer and elution solvent recommendations, technical notes, demonstration videos, and more!

2. WAIT For 5 minutes.

Novum™ SLE follows a simple 3 step procedure, allowing up to 96 samples to be processed in < 15 minutes. If further time savings are necessary, Novum SLE can be easily automated for rapid, hands free sample cleanup.

A Simplified Procedure

1. LOAD Dilute sample 1:1 with buffer or water and load onto NovumSLE sorbent using 2–15 seconds of vacuum.

novumsim

plified liquid extraction

novumsim


Optimized frit system provides a consistent

flow of sample into the Novum SLE sorbent

3. ELUTE Apply elution solvent and allow to elute via gravity. Complete elution with 10 seconds of vacuum.

novumsim


novumsim


Applying vacuum can reduce the amount

of time required for elution without

sacrificing cleanliness

See page 5 to learn moreabout the benefits of vacuum or

positive pressure manifold processing.

Novum Simplified Liquid Extraction (SLE)Estimated Time Required = <15 minutes

4

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0 %

10 %

20 %

30 %

40 %

50 %

60 %

70 %

80 %

90 %

100 %

Lyso 1 Lyso 2 PC 1 PC 2 PC 4

MTBE

DCM

Ethyl Acetate

A Cleaner Extract for Improved LC-MS/MS Analysis Phospholipids are present in all bioanalytical samples and can cause a variety of issues in LC-MS/MS analysis including ion suppression, reduced column lifetime, and reduced mass spec sensitivity.

Have you monitored the cleanup ability of your SLE product?

0 %

10 %

20 %

30 %

40 %

50 %

60 %

70 %

80 %

90 %

100 %


MTBE

DCM

Ethyl Acetate

Phospholipids remain in the

extracted sample

Phospholipid Breakthrough Using Various Extraction Solvents

Biotage® ISOLUTE® SLE+* (diatomaceous earth)

Novum SLE (synthetic SLE sorbent)

Novum SLEresults in cleaner

extracts

Biotage® ISOLUTE® SLE+* vs. Novum™ SLE

Lyso 1: 1-Palmitoyl-2-OH-sn-glycero-phosphocholine (m/z 496-184)Lyso 2: 1-Oleoyl-2-OH-sn-glycero-phosphocholine (m/z 522-184)PC 1: 1-Palmitoyl-2-Oleoyl-sn-glycero-phosphocholine (m/z 761-184)PC 2: 1-Stearoyl-2-Lindoleoyl-sn-glycero-phosphocholine (m/z 787-184)PC 4: 1-Oleoyl-2-Lindoleoyl-sn-glycero-phosphocholine (m/z 784-184)

Extraction Method1. Load diluted plasma (diluted 1:1

with water) onto a SLE 96-Well Plate, apply vacuum for 2-5 seconds.2. Allow sample to soak into the SLE sorbent for 5 minutes.3. Elute with ethyl acetate.

Plasma extractions were performed using Novum SLE MAX and Biotage® ISOLUTE® SLE+* 400 µL plates. The recommended protocol provided with each product was followed. Comparative separations may not be representative of all applications.

5

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0 %

10 %

20 %

30 %

40 %

50 %

60 %

70 %

80 %

90 %

100 %


Novum SLE

Biotage® Isolute® SLE+*

Avoid the Risks Associated with Diatomaceous Earth SLEDiatomaceous earth sorbents can have inconsistent aqueous holding capacities, which ultimately results in sample breakthrough before or during the final vacuum or positive pressure step. This will yield unwanted phospholipids in sample extract as seen below.

Novum SLE will not allow water and phospholipids to breakthrough the sorbent during a standard vacuum or positive pressure step due to the consistent aqueous holding capacity, allowing you to maximize your extraction without negatively affecting your clean-up.

Even under a standard

vacuum elution, Novum still removes

>99% of phospholipids

Plasma extractions were performed using Novum SLE MAX and Biotage® ISOLUTE® SLE+* 400 µL plates. The recommended protocol provided with each product was followed. Comparative separations may not be representative of all applications.

Phospholipid Breakthrough Using 5” Hg VacuumNovum SLE vs. Biotage® ISOLUTE® SLE+*



Extraction Method1. Load diluted plasma (diluted 1:1 with water)

onto a SLE 96-Well Plate, apply vacuum for 2-5 seconds.

2. Allow sample to soak into the SLE sorbent for 5 minutes.3. Elute with ethyl acetate.

Positive Pressure Manifold for Streamlined Sample Preparation • Pneumatic Handling • Consistent Flow Rates • Safe and Easy-to-Use • Go to www.phenomenex.com/Presston to learn more

Bitotage® ISOLUTE® SLE+*

Novum SLE

6

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More Reliable Clean-up,Lot-to-LotAs a naturally occurring product, the composition of diatomaceous earth used in traditional SLE products will vary. This results in inconsistency with the aqueous holding capacity which leads to incomplete cleanup of phospholipids.

Novum SLE is a lab manufactured sorbent, ensuring that the aqueous holding capacity is uniform leading to cleanup and recoveries that are consistent fromlot-to-lot.


Novum SLE Lot-to-Lot Phospholipid Breakthrough

Plasma extractions were performed using Novum SLE MINI and Biotage® ISOLUTE® SLE+* 200 µL plates. The recommended protocol provided with each product was followed. Comparative separations may not be representative of all applications.

0 %

10 %

20 %

30 %

40 %

50 %

60 %

70 %

80 %

90 %

100 %


Biotage® Isolute® SLE+* lot #13165345SA

Biotage® Isolute® SLE+* lot #12161236TA

Biotage® ISOLUTE® SLE+* Lot-to-Lot Phospholipid Breakthrough

0.000 %

0.002 %

0.004 %

0.006 %

0.008 %

0.010 %


Lot 1

Lot 2

Consistently remove > 99.9 % of phospholipids


Extraction Method1. Load diluted plasma (diluted 1:1 with water) onto a SLE 96-Well Plate, apply vacuum for 2-5 seconds.2. Allow sample to soak into the SLE sorbent for 5 minutes.3. Elute with ethyl acetate.

Wow!

Bitotage® ISOLUTE® SLE+* lot #12161236TA

Bitotage® ISOLUTE® SLE+ lot #13165345SA

7


View more applications and informationabout Novum PRO SLE at

www.phenomenex.com/NovumPRO

Get Down to the Lowest Extraction Levels with Novum PRO SLE Offers the same reliable synthetic sorbent as Novum with additional clean manufacturing steps to reach low levels of detection for sensitive MS applications, with the same quality reproducibility for high-throughput samples.

• Specific manufacture capabilities to improve matrix factor response and reduce noisy baselines for low level testing of biological samples

• API 6500+ fit for purpose testing to ensure clean baseline with each batch• Available in both MINI and MAX 96-well plate formats for high-throughput

applications

Simple Method with Easy Transferability• Load• Wait• Elute

Low Level Detection• Applications that require low levels of

detection and sensitivity can now be met by Novum PRO SLE

Equivalent reliability to traditional synthetic Novum SLE

Detection Limits Getting Smaller and Smaller?

2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6 3.8 4.0 min0.0

500.0

1000.0

1500.0

2000.0

2500.0

3000.0

3500.0

4000.0

4500.0

5000.0

5500.0

6000.0

6500.0

7000.0

7500.0

8000.0

8500.0

9000.0

9500.0

1.0e4

1.1e4

1.1e4

1.2e4

1.2e4

1.3e4

1.3e4

1.4e4

1.4e4

Intensity, cps

3.93

3.08

3.40

3.20

2.93

2.522.36

2.63

5 pg/mL (LLOQ) Estriol (E3)

App

ID 2

6234

Easy Method Development• Screen elution solvents in less time• Easily determine the best solvent to use

for clean backgrounds

8


Estrogen Panel from Plasma

Novum PRO SLE Protocol Pretreatment: 200 μL spiked plasma and 200 μL 50 mM Sodium phosphate buffer, pH unadjustedLoad: Pretreated sample onto Novum PRO MAX SLE (8E-S539-5GA), wait 5 minutes Elute: 2x 900 μL Hexane/Ethyl acetate (1:3)Dry: Down under NitrogenReconstitute: 50 μL Dansyl chloride/Sodium bicarbonate (1:1), incubate at 60 °C for 10 minutes

Low Level Hormones from Plasma Sensitive analysis requirements require clean MS backgrounds and precise low levels of analyte extractions. Novum PRO SLE makes this possible with LLOQs of 5-10 pg/mL for multiple estrogens and testosterone from plasma. With some diatomaceous earth SLE sorbents, background interferences can make peaks undetectable but with Novum PRO, recoveries are high with low limits of detection on a sensitive MS.

Reach highrecoveriesat LLOQ

Conditions for all samples:Columns: Kinetex® 1.7 μm Biphenyl

Dimensions: 50 x 2.1 mmPart No.: 00B-4628-AN

Mobile Phase: A: 0.1% Formic acid in Water B: 0.1% Formic acid in Methanol

Gradient: Time (min)012366.18

% B80809095958080

Flow Rate: 0.4 mL/minTemperature: Ambient

Injection: 10 µLDetection:

Analyte %Recovery %CV LLOQ Retention Time

Estrone (E1) 111 7 10 pg/mL 5.90

beta-Estadiol (E2) 92 7 10 pg/mL 4.35

Estriol (E3) 99 1 5 pg/mL 3.15

Estrogen Panel Recovery on Novum PRO SLE

Representative Chromatogram of Estrogen Panel at 2 ng/mL

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 min0.0

2.0e6

4.0e6

6.0e6

8.0e6

1.0e7

1.2e7

1.4e7

1.6e7

1.8e7

2.0e7

2.2e7

2.4e7

2.6e7

2.8e7

3.0e7

3.2e7

3.4e7

3.6e7

3.8e7

4.0e7

4.2e7

4.4e7

sp

c ,

yti

sn

etnI

App

ID 2

6214

New Novum PRO. Move Over Biotage® ISOLUTE® SLE+

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0.5

1

1.5

2

3

3.5

4

4.5

5

0 20 40 60 80 100

2.5

5.2

120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500

beta-E2 Calibration Curve 10 pg/mL (LLOQ) beta-Estradiol

R2=.9955

3.4 3.6 3.8 4.0 4.2 4.4 4.6 4.8 5.0 min0

200

400

600

800

1000

1200

1400

1600

1800

2000

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5600

5800

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6400

6600

6800

7000

7200

7400

7600

7800

8000

8200

8400

8600

8800

9000

Inte

nsity

, cps

4.28

4.08

3.92

3.47

4.65

4.73

5.15

App

ID 2

6233

App

ID 2

6230

App

ID 2

6234

2.2 2.4 2.6 2.8 3.0 3.2 3.4 3.6 3.8 4.0 min0.0

500.0

1000.0

1500.0

2000.0

2500.0

3000.0

3500.0

4000.0

4500.0

5000.0

5500.0

6000.0

6500.0

7000.0

7500.0

8000.0

8500.0

9000.0

9500.0

1.0e4

1.1e4

1.1e4

1.2e4

1.2e4

1.3e4

1.3e4

1.4e4

1.4e4

Intensity, cps

3.93

3.08

3.40

3.20

2.93

2.522.36

2.63

5 pg/mL (LLOQ) Estriol (E3) Calibration Curve of E3 from 2.5 to 500 pg/mL

0.2

0.6

1

0 20 40 60 80 100

1.2

120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 5000

0.4

0.8

1.4

1.6

1.8

2

2.2

2.4

App

ID 2

6231

R2=.9966

Move over Biotage! New Novum

PRO is here!

4.8 5.0 5.2 5.4 5.6 5.8 6.0 6.2 6.4 min0

200

400

600800

10001200

1400

16001800

20002200

2400

26002800

30003200

3400

36003800

40004200

4400

46004800

50005200

5400

56005800

60004.82

5.69

5.425.35

App

ID 2

6232

10 pg/mL (LLOQ) Estrone (E1) Calibration Curve of E1 from 5 to 500 pg/mL

0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500Analyte Conc. / IS Conc.

0.000.050.100.150.200.250.300.350.400.450.500.550.600.650.700.750.800.850.900.951.001.05

aerA

SI / aerA etylan

A

App

ID 2

6215

R2=.9963

Estrogen Panel from Plasma (cont’d)

Ana

lyte

Are

a /

IS A

rea

Analyte Conc. / IS Conc.

10

Ana

lyte

Are

a /

IS A

rea


Inte

nsit

y, C

PS

Inte

nsit

y, C

PS

Inte

nsit

y, C

PS


80 pg/mL Testosterone

1.9 2.0 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7 min0

100020003000400050006000700080009000

1e41.1e41.2e41.3e41.4e41.5e41.6e41.7e41.8e41.9e4

2e42.1e42.2e42.3e4

Inte

nsity

, cps

App

ID 2

6240

Reach required LLOQ with Novum PRO SLEwith no background interferences.

App

ID 2

6236

1000

19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 380

2000

3000

4000

5000

6000

7000

8000

9000

Plasma Blank

20 pg/mL of Testosterone

1.9 2.0 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7 min

1000

0

2000

3000

4000

5000

6000

7000

8000

9000

1e4

1.10e4

Inte

nsity

, cps

Testosterone Panel from Plasma Novum PRO SLE Protocol Pretreatment: 200 μL spiked plasma and 200 μL 50 mM Sodium phosphate buffer, pH unadjustedLoad: Pretreated sample onto Novum PRO MAX SLE (8E-S539-5GA), wait 5 minutes Elute: 2x 900 μL Hexane/Ethyl acetate (1:3)Dry: Down under NitrogenReconstitute: 50 μL Water/Methanol (80:20)

Conc. S/N

10 pg/mL 9.42

25 pg/mL 25.93

50 pg/mL 33.17100 pg/mL 77.59250 pg/mL 202.07500 pg/mL 138.62

Signal-to-Noise Ratio at Diverse Concentrations

10 pg/mL (LLOQ) of Testosterone

1.9 2.0 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7 min0200

1000

2000

3000

4000

5000

6000

7000

8000

9000

Inte

nsity

, cps

Conditions for all samples:Columns: Kinetex® 1.7 μm Biphenyl

Dimensions: 50 x 2.1 mmPart No.: 00B-4628-AN

Mobile Phase: A: 0.1% Formic acid in Water B: 0.1% Formic acid in Methanol

Gradient: Time (min)0233.15

% B6090956060

Flow Rate: 0.4 mL/minTemperature: Ambient

Injection: 10 µLDetection: SCIEX® 6500+ QTRAP® MS/MS

Sample: Testerstone

Inte

nsit

y, C

PS

Inte

nsit

y, C

PS

Inte

nsit

y, C

PS

Inte

nsit

y, C

PS

Good signal-to-noisefor different

concentrations oftestosterone!

1.9 2.0 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 3.1 3.2 3.3 3.4 3.5 3.6 3.7 min0200

1000

2000

3000

4000

5000

6000

7000

8000

9000

Inte

nsity

, cps

App

ID 2

6238

App

ID 2

6239

11


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Calibration Curve of Testosterone from 10 to 300 pg/mL

10 50 100 150 200 250Analyte Conc. / IS Conc.

0

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0.55

0.60

0.65

0.70

0.750.79

Ana

lyte

Are

a / I

S A

rea

App

ID 2

6235

Testerstone Recovery and at Diverse Concentrations

Testosterone % Recovery % CV

20 pg/mL 111 10

40 pg/mL 92 9

80 pg/mL 99 12150 pg/mL 105 12300 pg/mL 103 12

Recovery Data

R2=.995

Good recoveries and low %CVs at diverse levels of

concentration with clean-upusing Novum PRO!

Quotes, Methods, Tips... We're here to help

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Have Questions?

Testosterone Panel from Plasma (cont’d)

12

Ana

lyte

Are

a /

IS A

rea


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Faster Method DevelopmentSLE products may be the single easiest method development product, making it super easy to screen for diverse extraction solvents. With Novum™ SLE, you can trust that results will be consistent with the correct extraction solvent.

For typical applications, Novum™ SLE provides excellent recovery and cleanup using ethyl acetate as an extraction solvent, significantly reducing your method development time.

Recovery of 18 Pain Management Drugs from Urine

using a Single Extraction Method on Novum SLE

0 %

20 %

40 %

60 %

80 %

100 %

120 %

Amphe

tam

ine

Bupre

norp

hine

Codein

e

Fent

anyl

MDM

A

MDEA

Mep

eridine

Met

hadon

e

Mep

robam

ate

Met

ham

pheta

mine

Norfe

ntan

yl

Norm

eper

idinePCP

Propox

yphe

ne

Sufen

tanil

Tem

azep

am

Tram

adol

Nordiaz

epam

Rec

over

y (%

)

Analyte % RSD

Amphetamine 3

Buprenorphine 5

Codeine 10

Fentanyl 6

MDEA 4

MDMA 4

Meperidine 9

Meprobamate 7

Methadone 2

Methamphetamine 12

Nordiazepam 1

Norfentanyl 3

Normeperidine 4

PCP 2

Propoxyphene 9

Sufentanil 11

Temazepam 2

Tramadol 9

A Simplified Procedure

For typical applications ethyl acetate provides the

best balance of cleanup and recovery! But screening for multiple solvents is easy

with Novum SLE.Extraction Method1. Load diluted urine (diluted 1:1 with 0.5 M Ammonium hydroxide) onto Novum SLE MAX 96-well plate, apply vacuum for 2-15 seconds

2. Allow sample to soak into Novum SLE sorbent for 5 minutes

3. Elute with ethyl acetate

13


A Variety of Formats to Fit Your Sample and Throughput RequirementsTubesProcess samples as small as 100 µL or as large as 1 mL using Novum SLE tubes. Ideal for all types of applications including Bioanalytical, Food Safety, and Environmental.

96-Well PlatesProcess 96 samples at once in an easily automatable 96-well plate. Perfect for high-throughput applications

novumsim


novumsim


novumsim


novumsim


novumsim


novumsim


novumsim


novumsim


Novum SLE Tube 1 cc 3 cc 6 cc 12 cc

Maximum Sample Volume (before dilution)

100 µL 200 µL 500 µL 1 mL

Recommended Elution Volume 1.2 mL 1.8 mL 5 mL 10 mL

Novum SLE 96-Well Plate MINI MAX

Maximum Sample Volume (before dilution)

150 µL 200 µL

Recommended Elution Volume 1 mL 1.8 mL

New Novum PRO 96-Well Plates Made to reach low level extraction capabilities with new manufacturing

processes and same 96-well plate formats as traditional Novum forhigh-throughput, reliable SLE clean-up.

14


AccessoriesCollection Plates (deep well, polypropylene) UnitAH0-7192 96-Well Collection Plate, 350 µL/well Conical 50/pkAH0-7193 96-Well Collection Plate, 1 mL/well Conical 50/pkAH1-7022 96-Well Collection Plate, 2 mL/well Conical 50/pkAH0-8636 96-Well Collection Plate, 2 mL Round/Round, 8 mm 50/pkAH1-7025 96-Well Collection Plate, 1 mL/well Round, 7 mm 50/pkSealing MatsAH0-8597 Sealing Mats, Pierceable, 96-Square Well, Silicone 50/pkAH0-8598 Sealing Mats, Pre-Slit, 96-Square Well, Silicone 50/pkAH0-7362 Sealing Tape Pad 10/pkVacuum and Positive Pressure ManifoldsAH0-8950 96-Well Plate Manifold, Universal with Vacuum Gauge eaVM12 12-Position Vacuum Manifold Set eaVM24 24-Position Vacuum Manifold Set eaAH1-7033 Presston 1000 Postive Pressure Manifold, 96-Well Plate ea

Terms and Conditions Subject to Phenomenex Standard Terms and Conditions, which may be viewed at www.phenomenex.com/TermsAndConditions. Trademarks Novum, BE-HAPPY, and Presston are trademarks and Kinetex is a registered trademark of Phenomenex. Biotage and ISOLUTE are registered trademarks of Biotage AB.

Disclaimer Comparative separations may not be representative of all applications. Novum is patent pending. Phenomenex is in no way affiliated with Biotage AB.

FOR RESEARCH USE ONLY. Not for use in clinical diagnostic procedures.

© 2021 Phenomenex, Inc. All rights reserved.

TipFor more information about Phenomenex sample preparation products, visit www.phenomenex.com/sampleprepinfo

Redefine Your SLE Novum SLE 96-Well Plates

Part No. Description Unit8E-S138-FGA Novum SLE MINI 96-Well Plate 1/pk8E-S138-5GA Novum SLE MAX 96-Well Plate 1/pk8E-S539-FGA Novum PRO SLE MINI, 96-Well Plate 1/pk8E-S539-5GA Novum PRO SLE MAX, 96-Well Plate 1/pk

Novum SLE Tubes

Part No. Description Unit8B-S138-FAK Novum SLE 1 cc tubes 100/pk8B-S138-5BJ Novum SLE 3 cc tubes 50/pk8B-S138-JCH Novum SLE 6 cc tubes 30/pk8B-S138-KDG Novum SLE 12 cc tubes 20/pk

Phenomenex warrants the Presston 1000 Positive Pressure Manifold against defects in materials and workmanship under normal installation, use, and maintenance for a period of 12 months following delivery. Please visit www.phenomenex.com/presstonwarranty for complete warranty information.

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www.phenomenex.comPhenomenex products are available worldwide. For the distributor in your country, contact Phenomenex USA, International Department at [email protected]

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