Supported Liquid Extraction (SLE) · SLE Protocol Packed with diatomaceous earth, Strata DE is a...

Supported Liquid Extraction (SLE)For Clinical Research Applications

Novum™ Synthetic SLEUnique, Synthetic Sorbent that Provides Consistent, Reliable Results

www.phenomenex.com/ClinSLE

Strata® DE Diatomaceous Earth SLECost-Effective, Guaranteed Alternative to Traditional Diatomaceous Earth SLE Sorbents

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Phenomenex l WEB: www.phenomenex.com2

Supported Liquid Extraction

Synthetic Diatomaceous Earth

Lot-to-lot consistency and reproducibility

Cost effective and large volume capabilities

MINI 96-Well Plates, MAX 96-Well Plates

200 µL 96-Well Plates, 400 µL 96-Well Plates

Ethyl Acetate, Methyl Tert-Butyl Ether (MTBE)

Dichloromethane (DCM), Hexane, MTBE, Ethyl Acetate

1 cc, 3 cc, 6 cc, 12 cc 12 cc and 60 cc

Sorbent

Extraction Solvents

Advantages

Plate Formats

Tube Formats

simplified liquid extraction

Choose the Best SLE Solution for Your Analysis

Phenomenex provides two options for your SLE analysis, a unique synthetic sorbent and a traditional diatomaceous earth sorbent, each of which provides its own benefits. Select the best option for your work by using the chart below.

Call us

www.phenomenex.com/SPE

Need Help Selecting the Right SLE Option?

Live Chat www.phenomenex.com/LiveChat

or

Phenomenex l WEB: www.phenomenex.com 3

Explore Your SLE Options and Applications

Novum™ SLE

Unique, Synthetic Sorbent ......................................................p. 4

Strata® Diatomaceous Earth (DE) SLECost-Effective Guaranteed Alternative Sorbent ...........p. 5

Applications25-OH Vitamin D2/D3 ...............................................................p. 6NSAIDs .............................................................................................p. 7Corticosteroids .............................................................................p. 8Nicotine and Metabolites .........................................................p. 9THC-COOH and Barbiturates ...............................................p. 10Ordering Information .................................................................p. 11

novumsim

plified liquid extraction

If Phenomenex SLE products do not perform as well or better than your current SLE product, return the product with comparative data within 45 days for a FULL REFUND.


Synthetic SLE

Lyso 1 Lyso 2 PC 1 PC 2 PC 40.000 %

0.002 %

0.004 %

0.006 %

0.008 %

0.010 %

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)

As a unique, synthetic SLE sorbent, expect Novum to provide reliable, more consistent lot-to-lot cleanup that excels under extraction conditions where traditional diatomaceous earth SLE falls short, such as ethyl acetate.Recommended Uses:• Ethyl acetate extractions

• Small volume samples

Lot–to-Lot Phospholipid Breakthrough: Novum SLE vs. Traditional Diatomaceous Earth SLE

Biotage ISOLUTE SLE+ Novum SLE

0 %

10 %

20 %

30 %

40 %

50 %

60 %

70 %

80 %

90 %

100 %

Lyso 1 Lyso 2 PC 1 PC 2 PC 4

Consistently Remove More Phospholipids

than Biotage ISOLUTE SLE+

simplified liquid extraction

0 %

10 %

20 %

30 %

40 %

50 %

60 %

70 %

80 %

90 %

100 %

Lyso 1 Lyso 2 PC 1 PC 2 PC 4

Biotage® ISOLUTE® SLE+ lot #13165345SA

Biotage ISOLUTE SLE+ lot #12161236TA

Novum Lot 1

Novum Lot 2

Comparative separations may not be representative of all applications.


Diatomaceous Earth SLE

96-Well Plate:Strata DE SLE 400 µL 96-Well Plate Biotage ISOLUTE SLE+ 400 µL 96-Well Plate

Part No.: 8E-S325-5GB (Strata DE)

Load:300 µL pre-treated urine sample onto plate (apply vacuum or positive pressure to pull/push sample into sorbent if necessary)

Wait: 6 minutes

Elute: 3x 600 µL Dichloromethane/IPA (95:5)

Apply: Vacuum or apply positive pressure at 5-10” Hg for 10 seconds

Dry: Sample under slow stream of Nitrogen at 30°C

Reconstitute:100 µL 0.1% Formic Acid/Methanol (4:1) with internal standard

SLE Protocol

Packed with diatomaceous earth, Strata DE is a cost-effective alternative to traditional SLE products such as Biotage® ISOLUTE® SLE+, Thermo Fisher® Hypersep™ SLE, and Agilent® Chem Elut™ SLE that won’t require you to sacrifice your results.Recommended Uses:• Direct alternative to traditional diatomaceous earth SLE sorbents

• Non-oxygenated extraction solvents such as, DCM and Hexane

Achieve the Same Recoveries as Biotage ISOLUTE SLE+,

at a Lower Price!

Strata DEBiotage

ISOLUTE SLE+

Analyte% Recovery

% CV (n=8)

% Recovery% CV (n=8)

6-MAM 98 9 88 16

Alprazolam 104 10 98 11

Benzoylecgonine 88 6 98 11

Buprenorphine 93 7 102 15

Codeine 99 12 93 9

Diazepam 107 7 104 6

Fentanyl 85 5 94 8

Hydrocodone 104 11 93 11

Hydromorphone 95 9 93 11

Lorazepam 94 8 98 8

Methamphetamine 92 16 102 8

Morphine 98 12 94 12

Norbuprenorphine 101 11 92 11

Nordiazepam 100 9 92 8

Norfentanyl 113 7 110 11

Oxycodone 97 5 93 11

PCP 90 7 98 6

Recovery Values and % CVs: Strata DE SLE vs. Biotage ISOLUTE SLE+

Supported Liquid Extraction

Comparative separations may not be representative of all applications.

• High Recoveries• Low RSDs


25-OH Vitamin D2/D3 from Plasma

96-Well Plate: Strata® DE 200 µL 96-Well Plate

Part No.: 8E-S325-FGB

Load:200 µL Pre-treated sample onto plate (apply vacuum or positive pressure to pull/push sample into sorbent)

Wait: 5 minutes

Elute: 2x 600 µL Heptane

Apply:Vacuum or apply positive pressure at 5-10” Hg for 10 seconds

Dry: Sample under slow stream of Nitrogen at 30 °C

Reconstitute:100 µL Methanol/Water (80:20) with internal standard

LC-MS/MS ConditionsColumn: Kinetex® 2.6 µm EVO C18

Dimensions: 50 x 2.1 mmPart No.: 00B-4725-AN

Guard Cartridge: SecurityGuard™ ULTRA EVO C18Guard Part No.: AJ0-9298Mobile Phase: A: 0.1 % Formic acid in Water

B: 0.1 % Formic acid in AcetonitrileGradient: Time (min) % B

0 600.83 953.33 953.34 605.83 60

Flow Rate: 0.5 mL/minInjection Volume: 20 µL

Detection: MS/MS (SCIEX API 4000™), ESI+

Pre-treatmentDilute 100 µL of spiked plasma (100 ng/mL) with 100 µL of IPA/Water (1:1).

SLE Protocol

25-OH Vitamin D2 25-OH Vitamin D3

% Recovery 96 84

% RSD (n=8) 9 9

High Recoveries Using Strata DE SLE

0.5

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Extracted Chromatogram for 25-OH-Vitamin D2/D3

®®®®

Sample: 1. 25-OH-Vitamin D32. 25-OH-Vitamin D2

LC-MS/MS ConditionsColumns: Kinetex® 2.6 µm C18

Dimensions: 50 x 2.1 mmPart No.: 00B-4462-AN

Guard Cartridge: SecurityGuard™ ULTRA C18Part No.: AJ0-8782

Mobile Phase: A: 0.1 % Formic acid in WaterB: 0.1 % Formic acid in Acetonitrile

Gradient: Time (min) % B0 102.5 1003.5 1003.51 10

Flow Rate: 0.5 mL/min Injection: 3 µL

Detection: MS/MS (SCIEX API 4000™)

• Small Sample Volume• High Recovery for NSAIDs• Cost Effective NSAID Extraction


NSAIDs from Urine

Pre-treatmentDilute 100 µL of spike urine (100 ng/mL) with 100 µL of 0.1 % formic acid.

SLE Protocol

96-Well Plate: Strata® DE 200 µL

Part No.: 8E-S325-FGB

Load: 200 µL Pre-treated sample

Wait: 6 minutes

Elute: 2x 600 µL Ethyl acetate/Hexane (3:1)


Reconstitute:100 µL 0.1 % Formic acid in Methanol with 100 ng/mL concentration of Diclofenac D4, Acetaminophen D7, Naproxen D3

Acetaminophen Naproxen Diclofenac

% Recovery 96 84 109

% RSD (n=8) 9 9 5

Recovery Values

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 min0.0

5.0e4

1.0e5

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4.5e5

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8.0e5

8.5e58.8e5

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TIC for Extracted Sample

Sample: 1. Acetaminophen2. Naproxen3. Diclofenac


Corticosteroids from Plasma

Pre-treatmentDilute 100 µL of spiked plasma (125 ng/mL) with 200 µL of Water

SLE Protocol96-Well Plate: Strata® DE 400 µL

Part No.: 8E-S325-5GB

300 µL treated sample onto plate (apply vacuum or positive pressure to pull/push sample into sorbent) if neccessary

Wait: 5 minutes

Elute:3x 600 µL Dichloromethane (DCM) or 3x 600 µL Ethyl Acetate

Apply: Vacuum or apply positive pressure at 5-10” Hg for 10 seconds


Reconstitute: 200 µL Acetonitrile/Water (20:80)

Achieve High Recoveries Across Multiple Elution Solvents

Elution Solvent: Dichloromethane Ethyl Acetate

% Recovery% RSD (n=4)

% Recovery% RSD (n=10)

ß-Methasone 92 4 98 6

Cortisone 96 10 96 8

Coritcosterone 92 3 74 10

Cortisone Acetate 90 12 112 12

Triamcinolone 13 8 92 9

Prednisone 94 7 93 10

Testosterone 95 5

*Testosterone was not extracted using Ethyl Acetate

LC-MS/MS ConditionsColumn: Kinetex® 2.6 µm C18

Dimensions: 50 x 2.1mmPart No.: 00B-4462-AN

Guard Cartridge: SecurityGuard™ ULTRA C18Guard Part No.: AJ0-8782Mobile Phase: A: 0.1 % Formic acid in water

B: 0.1 % Formic acid in AcetonitrileGradient: Time (min) B (%)

0 203 953.5 953.51 206 20

Flow Rates 0.5 mL/min

Injection Volume: 5 µLDetection: MS/MS (SCIEX API 4000™), ESI+

• Extraction of a wide range of corticosteroids

• Two extraction solvents for diverse options

Load:

Hundreds of Sample Preparation Resources•Brochures,Technical Notes,Webinars and more!

Visit www.phenomenex.com/sampleprep

•Consistently good recovery across QC•Low Variation

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Table 1. Mass Transitions

Compounds Q1 Mass (amu)

Q3 Mass (amu)

Dwell Time (amu) DP CE

Anabasine 1* 163 80 25 80 27

Anabasine 2 163 120.1 25 80 22

Anabasine D4 1* 167 84 25 80 27

Anabasine D4 2 167 124 25 80 22

Cotinine 1 177 80 25 80 29

Cotinine 2* 177 98 25 70 28

Cotinine D3 1* 180 80 25 80 28

Cotinine D3 2 180 101 25 80 28

3-Hydroxycotinine 1* 193 80 25 60 34

3-Hydroxycotinine 2 193 134 25 60 25

3-Hydorxycotinine D3 1* 196 80 25 60 34

3-Hydroxycotinine D3 2 196 134 25 60 25

Nicotine 1 163 130 25 60 26

Nicotine 2* 163 132 25 60 20

Nicotine D4 1* 167 134 25 60 26

Nicotine D4 2 167 136 25 60 20

Nornicotine 1 149 80 25 50 25

Nornicotine 2* 149 130 25 50 21

Nornicotine D4 1 153 84 25 55 29

Nornicotine D4 2* 153 134 25 55 18

Anabasine 1* 163 80 25 80 27

* Quantitation mass

Table 2. Novum Recovery

Anabasine Nicotine Nornicotine Cotinine 3-OH-Cotinine

QCL 71.1 82.8 62.4 96.2 73.0

QCM 79.5 83.9 64.3 95.6 77.5

QCH 83.5 89.6 70.4 91.0 76.7

Figure 1. Representative chromatograms of QCM at 200 ng/mL in human urine

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

1

2

3

4

5

min0.0

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4.0E5

6.0E5

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1.0E6

1.2E6

1.4E6

1.6E6

1.8E6

2.0E6

2.2E6

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2.6E6

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3.0E6

3.2E6

3.4E6

3.6E6

3.8E6

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Ap

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87

1. 3-Hydroxycotinine

2. Nornicotine

3. Cotinine

4. Anabasine

5. Nicotine

Ap

p ID

237

87

Pre-treatmentTo 100 µL urine add 200 µL of 2% Ammonium hydroxide in water and 50 µL of mixed deuterated internal standards, mix.

SLE Protocol

Nicotine and Metabolites from Urine

Novum Recovery

LC-MS/MS Conditions Column: Kinetex® 2.6 µm EVO C18

Dimensions: 100 x 3.0 mmPart No.: 00D-4725-Y0

Guard Cartridge: SecurityGuard™ ULTRA EVO C18Guard Part No.: AJ0-9287Mobile Phase: A: 20 mM Ammonium Bicarbonate, pH 8.2

B: MethanolGradient: Time (min) B (%)

0 103 905 905.01 106. 10

Flow Rate: 0.75 mL/minTemperature: Ambient

Detection: MS/MS SCIEX QTRAP 4500 Turbo™)

ESI Ionization Source ParametersCurtain Gas (CUR): 20

Collision Gas (CAD): 7 Temperature (TEM): 600

Gas 1 (GS1): 50Gas 2 (GS2): 50

IS: 4500Entrance Potential (EP): 10

Collision Potential (CXP): 12

Representative Chromatograms of QCM at 200 ng/mL in Human Urine

96-Well Plate: Novum™ SLE MAX

Part No.: 8E-S138-5GA

Load: Pre-treated sample and pulse vacuum (~5” Hg) for 5-10 seconds or until sample has completely entered the sorben

Wait: 5 minutes

Elute: 3 x 600 µL 1% Formic acid in DCM/IPA (95:5)

Apply: Vacuum or apply positive pressure at 5” Hg for 15 seconds to complete elution

Dry: Sample under slow stream of Nitrogen at 45 ˚C

Reconstitute: In 300 µL 20 mM Ammonium bicarbonate (pH 8.2)/Methanol (90:10), vortex

1. 3-Hydroxycotinine

2. Nornicotine

3. Cotinine

4. Anabasine

5. Nicotine

Anabasine Nicotine Nornicotine Cotinine 3-OH-Cotinine

Quality Control

Low 71.1 82.8 62.4 96.2 73.0

Quality Control

Medium 79.5 83.9 64.3 95.6 77.5

Quality Control

High 83.5 89.6 70.4 91.0 76.7

0.0

20.010.0

40.030.0

60.050.0

80.070.0

100.090.0

Sample:

• High and consistent recoveries using Hexane/Ethyl Acetate (3:1) as extraction solvent

• Good sensivitity of THC-COOH

novumsim



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 8.0 min0.0

5000.0

1.0e4

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LC-MS/MS ConditionsAnalytical Column: Kinetex® 2.6 µm EVO C18 100Å

Dimension: 50 x 2.1 mmPart No.: 00B-4725-AN

Guard Cartridge: SecurityGuard™ ULTRA C18Guard Part No.: AJ0-7844Mobile Phase: A: 10 mM Ammonium bicarbonate, pH 9

B: AcetonitrileGradient: Time (min) B (%)

0 52 155 205.01 606 606.1 57.5 5

Flow Rate: 0.5 mL/minTemperature: Room temperature

Injection Volume: 2 µLInstrument: Agilent® 1200 LC System

Detection: MS/MS (SCIEX API 4000™), ESI+

THC-COOH and Barbiturates from Urine

96-Well Plate: Novum™ SLE MAX

Part No.: 8E-S138-5GA

Load: Pretreated sample and pulse vacuum (~5” Hg) for 5-10 seconds or until sample has completely entered the sorbent

Wait: 5 minutes

Elute: 2x 900 µL Hexane/Ethyl acetate (3:1)

Apply: Vacuum or apply positive pressure at 5” Hg for 15 seconds to complete elution

Dry: Sample under slow stream of Nitrogen at 30 ˚C

Reconstitute: In 200 µL of Methanol/Water (50:50) containing 1 % Ammonium hydroxide and 100 ng/mL of COOH-THC-D3 and 250 ng/mL of Pentobarbital-D5

% Recovery% CV (N=4)

Amobarbital 93 4

Pentobarbital 87 4

Butabital 93 7

Phenobarbital 85 8

Secobarbital 88 13

THC-COOH 81 12

Recovery values of Barbiturates and THC-COOH for Hexane/EtOAc (3:1)

Representative chromatogram for analytes extracted using Hexane/EtOAc (3:1)

Pre-treatmentTo 150 µL urine add 25 µL β-Glucuronidase with 73 µL of 0.1M Ammonium acetate buffer (pH 4.0). Combine with 200 µL of 1% Formic acid in water.

SLE Protocol

Clean-up with Novum SLE

BEFORETurbid

and Dirty

AFTERConcentrated and Clean

Sample:

1. Phenobarbital

2. Butabital

3. Pentobarbital

4. Amibarbital

5. Secobarbital

6. THC-COOH

1

2

3

45

6

Vacuum ManifoldsVacuum ManfoldsPart No. Description UnitAH0-6023 12-Position Tube Vacuum Manifold Set eaAH0-6024 24-Position Tube Vacuum Manifold Set eaAH0-8950 96-Well Plate Manifold, Universal with Vacuum Gauge ea

Phenomenex warrants that for a period of 12 months following delivery, the Presston 100 Positive Pressure Manifold you have purchased will perform in accordance with the published specifications and will be free from defects in materials or workmanship. In the event that the Presston 100 Positive Pressure Manifold does not meet this warranty, Phenomenex will repair or replace defective parts. Please visit www.phenomenex.com/Presston for complete warranty information.

Presston 100 Positive Pressure ManifoldPresston 100Part No. Description UnitAH0-9334 Presston 100 Positive Pressure Manifold, 96-Well Plate eaAH0-9342 Presston 100 Positive Pressure Manifold, 1 mL Tube Complete Assembly eaAH0-9347 Presston 100 Positive Pressure Manifold, 3 mL Tube Complete Assembly eaAH0-9343 Presston 100 Positive Pressure Manifold, 6 mL Tube Complete Assembly ea

The Presston 100 96-Well Positive Pressure Manifold can also process 1, 3, and 6 mL tubes using the following adapter kits: Presston 100 Tube Adapter Kits (for AH0-9334)Part No. Description UnitAH0-9344 1 mL Tube Adapter Kit eaAH0-9345 3 mL Tube Adapter Kit eaAH0-9346 6 mL Tube Adapter Kit ea

Synthetic SLENovum™ 96-Well PlatesNovum Simplified Liquid Extraction SLE Well PlatesPart No. Description Unit8E-S138-FGA Novum SLE MINI 96-Well Plate 1/pk8E-S138-5GA Novum SLE MAX 96-Well Plate 1/pk

Novum TubesNovum Simplified Liquid Extraction SLE TubesPart 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

Diatomaceous Earth SLE

Strata DE TubesStrata DE Diatomaceous Earth SLE TubesPart No. Description Unit8B-S325-KDG Strata DE SLE 12 cc Tubes 20/pk8B-S325-VFF Strata DE SLE 60 cc Tubes 16/pk

Strata® DE 96- Well Plates Strata DE Diatomaceous Earth SLE Well PlatesPart No. Description Unit8E-S325-FGB Strata DE SLE 200 µL 96-Well Plate 2/pk8E-S325-5GB Strata DE SLE 400 µL 96-Well Plate 2/pk

If Phenomenex SLE products do not perform as well or better than your current SLE product, return the product with comparative data within 45 days for a FULL REFUND.


Start Your New SLE Method

Terms and ConditionsSubject to Phenomenex Standard Terms and Conditions, which may be viewed at www.phenomenex.com.TrademarksStrata and Kinetex are registered trademarks and Novum, Presston, and SecurityGuard are trademarks of Phenomenex. Biotage and ISOLUTE are registered trademarks of Biotage AB Corp. Thermo Fisher and HyperSep is a trademark of Thermo Fisher Scientific. Agilents is a registered trademark and ChemElut is a trademark of Agilent Technologies, Inc. QTRAP 4500 Turbo is a registered trademark and API 4000 is a trademark of AB SCIEX Pte. Ltd. AB SCIEX is being used under license.DisclaimerPhenomenex is not affiliated with Biotage AB Corp., Thermo Fisher Scientific, or Agilent Technologies, Inc.FOR RESEARCH USE ONLY. Not for use in diagnostic procedures. Novum is patent pending.Comparative separations may not be representative of all applications.

© 2018 Phenomenex, Inc.All rights reserved.

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Supported Liquid Extraction (SLE)For Clinical Research Applications

Supported Liquid Extraction (SLE) · SLE Protocol Packed with diatomaceous earth, Strata DE is a...

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