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
Rev
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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?
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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.
Phenomenex l WEB: www.phenomenex.com4
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.
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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
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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
1
2
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 min0.0
1000.0
2000.0
3000.0
4000.0
5000.0
6000.0
7000.0
8000.0
9000.0
1.0e4
1.1e4
1.2e4
1.3e4
1.4e4
1.5e4
1.6e4
1.7e4
1.8e4
1.9e4
Inte
nsity
, cp
s
Ap
p ID
245
52
Inte
nsity
, cp
s
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
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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)
Dry: Sample under slow stream of Nitrogen at 30 °C
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
1.5e5
2.0e5
2.5e5
3.0e5
3.5e5
4.0e5
4.5e5
5.0e5
5.5e5
6.0e5
6.5e5
7.0e5
7.5e5
8.0e5
8.5e58.8e5
Inte
nsity
, cp
sIn
tens
ity, c
ps
Ap
p ID
246
14
TIC for Extracted Sample
Sample: 1. Acetaminophen2. Naproxen3. Diclofenac
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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
Dry: Sample under slow stream of Nitrogen at 30 °C
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
novumsim
plified liquid extraction
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TN-0103
Page 2 of 8
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
2.0E5
4.0E5
6.0E5
8.0E5
1.0E6
1.2E6
1.4E6
1.6E6
1.8E6
2.0E6
2.2E6
2.4E6
2.6E6
2.8E6
3.0E6
3.2E6
3.4E6
3.6E6
3.8E6
4.0E6
INT
EN
SIT
Y, C
PS
Ap
p ID
237
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
plified liquid extraction
Phenomenex l WEB: www.phenomenex.com10
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
1.5e4
2.0e4
2.5e4
3.0e4
3.5e4
4.0e4
4.5e4
5.0e4
5.5e4
Inte
nsity
, cps
Inte
nsity
, cp
s
Ap
p ID
241
26
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.
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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.
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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Supported Liquid Extraction (SLE)For Clinical Research Applications