Simplify Your Liquid Extractions

Transcription

1 a NEW way to Simplify Your Liquid Extractions Faster and easier than liquid-liquid extraction Easily automate to increase throughput Avoid analyte loss due to emulsions Process small and large sample volumes Are You Currently Using SLE? Learn why Novum is redefining SLE Dinosaur

2 Simplify Your Liquid-Liquid Extraction Now you can alleviate the challenges associated with traditional liquid-liquid extraction with Novum Simplified Liquid Extraction (SLE) (patent pending). A Simple, Automatable Procedure 1. Load Your Sample in aqueous solvent Instantly Increase Throughput Novum SLE 96-well plates and tubes can be processed using vacuum manifolds or automated liquid handlers. No emulsions 2. Collect Your Target Analytes in water immiscible solvent Interferences (i.e. phospholipids, proteins, salts, etc.) Target Analytes No need to manually separate liquid phases 215 Phenomenex, Inc. All rights reserved. 2

3 A Faster and Easier Solution... p. 4 Consistent, Reliable Results... p. 5 Easily Clean Up Urine Samples... p. 6 Automate Plasma Cleanups... p. 7 Tackle Complex Food Matrices... p. 8 Clean Up Challenging Personal Care Products... p. 9 Multiple Formats for Method Flexibility... p. 1 Ordering Information... p. 11 Are You Currently Using SLE? Find out what makes Novum SLE different at /Dinosaur Terrellasaurus first discovered in Torrance, CA. Phenomenex l WEB: 3

4 Faster and Easier than Liquid-Liquid Extraction Novum SLE will instantly increase your throughput by eliminating time consuming steps and reducing the risk of analyte loss. If further time savings are necessary, Novum SLE can be easily automated for rapid, hands free sample cleanup. Traditional Novum Simplified Liquid-Liquid Extraction1 Estimated Time Required = 25 minutes Liquid Extraction (SLE) Estimated Time Required = <15 minutes 1. Dilute sample 1:1 with buffer or 1. Dilute sample 1:1 with buffer or water 2. Mix for 1 minutes 2. Wait 5 minutes 3. Centrifuge for 1 minutes 3. Apply elution solvent and allow to water and add extraction solvent and load onto Novum SLE sorbent using 2 15 seconds of vacuum elute via gravity. Complete elution with 1 seconds of vacuum. 4. Pour off or freeze supernatant Rapid, automatable method for high throughput cleanup Stop worrying about analyte loss due to emulsions Visit for buffer and elution solvent recommendations, technical notes, demonstration videos, and more! 1. Russell Grant, Matthew Crawford, Brian Rappold, and Stacy Dee. Errors in Bioanalysis Due to Phospholipids Definitive Measurement, Mechanism, and Management. ASMS

5 Reliable Cleanup, from Lot-to-Lot Avoid inconsistent results due to the formation of emulsions or variances between technicians. The unique, synthetic Novum SLE sorbent is designed to provide consistent cleanup and recoveries from lot-to-lot. % Recovery of Corticosteroids from Plasma Across 3 Different Novum Sorbent Lots Achieve consistent recoveries from lot-to-lot 14 % 12 % Lot 1 Lot 2 Lot 3 1 % Recovery (%) 8 % 6 % 4 % 2 % % 11a-OH-progesterone Corticosterone Cortisone Cortisone Acetate Prednisone Testosterone Triamcinolone Acetonide Analyte % RSD 11a-OH-progesterone 3.9 Corticosterone 2.5 Cortisone 2.8 Cortisone Acetate 5.8 Prednisone 3.7 Testosterone 5.2 Triamcinolone Acetonide 3.4 Extraction Method 1. Load diluted plasma (diluted 1:1 with water) 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 Phenomenex l WEB: 5

6 Easily Clean Up Urine Samples Urine contains salts and other contaminants that can decrease your column lifetime and interfere with MS detection. Rapidly and reproducibly cleanup diluted urine or hydrolyzed urine samples without the need to perform extensive method development. COOH-THC (11-nor-9-carboxy-Δ 9 -tetrahydrocannabinoid) from Urine using Novum SLE 1.25e5 1.2e5 Enzymatic Hydrolysis Intensity, cps 1.1e5 1.e5 9.e4 8.e4 7.e4 6.e4 5.e4 4.e4 3.e4 2.e4 1.e HPLC Conditions Column: Kinetex 2.6 µm C8 Dimensions: 5 x 2.1 mm Part No.: B-4497-AN Mobile Phase: A:.1 % Formic acid in Water B: Methanol/Acetonitrile (5:5) Gradient: Time (min) % B Flow Rate:.5 ml/min Injection Volume: 2 µl Temperature: Ambient Detection: MS/MS, API 5 (AB SCIEX), ESI+ App ID min To 3 μl urine add 75 μl of 3 mm Ammonium acetate (ph 4.) and 25 μl of β-glucuronidase solution (1, units/ml). Mix and vortex for 3 seconds. Incubate at 37 4 C for 6 minutes. After incubation, bring samples to room temperature. Novum SLE Procedure Load pre-treated sample onto a Novum SLE MAX 96-well plate. Apply a gentle pulse of vacuum (~5-1 seconds at 5 Hg or less) until the sample has completely entered the media. Wait for 5 minutes. Dispense 2x 9 μl Ethyl acetate onto the Novum SLE sorbent and collect solvent under gravity in a collection plate, pooling fractions. Apply vacuum at 5 Hg (or lower) for 2-3 seconds to complete the extraction. Dry down and reconstitute in 2 μl mobile phase. MS/MS Conditions CUR: 25. GS1: 5. GS2: 5. TEM: 6. CAD: 7. IS: 55. DP: 55. Analyte 11-nor-9-carboxy- Δ 9 -THC % Absolute Recovery %CV (N=8)

7 Rapid, Automatable Cleanup of Plasma Samples Plasma contains a variety of interferences that can negatively affect downstream analysis. Proteins can clog HPLC columns while phospholipids can cause ion suppression or enhancement in MS analysis. Novum SLE provides a simple, automatable solution for removing these interferences. Cortisone and Prednisolone from Plasma using Novum SLE on a Tecan Freedom EVO Liquid Handler Intensity, cps 1.5e5 1.4e5 1.3e5 1.2e5 1.1e5 1.e5 9.e4 8.e4 7.e4 6.e4 5.e4 4.e4 3.e4 2.e4 1.e4. Cortisone HPLC Conditions Column: Kinetex 2.6 μm Biphenyl Dimensions: 5 x 3. mm Part No.: B-4622-Y Mobile Phase: A: 1 mm Ammonium acetate in Water B: 1 mm Ammonium acetate in Methanol Gradient: Time (min) % B Flow Rate:.45 ml/min Injection Volume: 1 µl Temperature: Ambient Detection: MS/MS, API 5 (AB SCIEX), ESI+ Cortisol-D4 (IS) Prednisolone min App ID Sample Pre-treatment Dilute 15 μl plasma with 15 μl 5 mm Sodium phosphate dibasic heptahydrate (ph unadjusted). Mix briefly (3-5 seconds). Novum SLE Procedure Load pre-treated sample onto a Novum SLE MINI 96-well plate. Apply a gentle pulse of vacuum (~5-1 Hg for 2 seconds) until the sample has completely entered the media. Wait for 5 minutes. Dispense 1 ml Ethyl acetate onto the Novum SLE sorbent and collect solvent under gravity in a collection plate. Apply vacuum at 5 Hg (or lower) for 45 seconds to complete the extraction. Dry down and reconstitute in 15 μl mobile phase fortified with cortisol-d4. Analyte RT, min Q1, Da Q3, Da CE Cortisone Cortisol-D4 (IS) Prednisolone Sample pre-treatment and Novum SLE procedure were performed using a Tecan Freedom EVO liquid handler to provide rapid, hands free sample processing. View the complete technical note at Analyte % Absolute Recovery %CV (N=8) Prednisolone Cortisone Phenomenex l WEB: 7

8 Remove Interferences from Complex Food Matrices Food matrices are complex and contain a variety of interferences that can negatively impact downstream analysis. Liquid-liquid extraction has historically been a popular means of sample cleanup for food matrices however this process can be laborious. Now labs can simplify the liquid extraction process with Novum SLE. Acrylamide from Coffee using Novum SLE 1.3e6 Sample Pre-treatment Intensity, cps 1.2e6 1.1e6 1.e6 9.e5 8.e5 7.e5 6.e5 5.e5 4.e5 3.e5 2.e5 1.e5 Acrylamide Acrylamide 13 C 3 App ID Prepare ground coffee as directed on the coffee product using 1,5 ml of boiling water and 6 g of ground coffee. Allow to reach room temperature before further analysis. To 8 µl of the prepared coffee add 15 µl 2 % Ammonium hydroxide in water. Vortex for 3 seconds. Before After min HPLC Conditions Column: Synergi 4 µm Hydro-RP Dimensions: 5 x 2. mm Part No.: B-4375-B Mobile Phase: A:.1 % Formic acid in Water B:.1 % Formic acid in Methanol Gradient: Time (min) % B 1 1 Flow Rate: 3 µl/min Injection Volume: 5 µl Temperature: Ambient Detection: MS/MS, API 5 (AB SCIEX), ESI+ Analyte RT, min Q1, Da Q3, Da CE Acrylamide Acrylamide 13 C 3 (IS) Analyte % Absolute Recovery %CV (N=6) Acrylamide (1 ng/ml) Novum SLE Procedure Load pre-treated sample onto a Novum SLE 6 cc tube. Apply a short and gentle pulse of vacuum (~5-1 Hg) until the sample has completely entered the media. Wait for 5 minutes. Dispense 2x 2.5 ml Ethyl acetate/tetrahydrofuran (1:1) onto the Novum SLE sorbent and collect solvent under gravity into a collection tube that contains 1 µl Ethylene glycol. Apply vacuum at 5 Hg (or lower) for 2-3 seconds to complete the extraction. Dry down and reconstitute in 3 µl Water. View the complete technical note at 8

9 Clean Up Challenging Personal Care Products Personal care products can pose several challenges during the sample preparation process including analyte loss due to the formation of emulsions that can occur during lengthy liquidliquid extractions. Eliminate the risk of analyte loss due to emulsions and simplify the liquid extraction process with Novum SLE. Parabens from Baby Wash and Shampoo using Novum SLE mau Methyl Paraben Ethyl Paraben Propyl Paraben Butyl Paraben HPLC Conditions Column: Kinetex 5 µm EVO C18 Dimensions: 1 x 3. mm Part No.: D-4633-Y Mobile Phase: A: 1 % Acetonitrile in Water B: Acetonitrile Gradient: Time (min) Flow Rate: 1.5 ml/min Injection Volume: 2 µl Temperature: Ambient Detection: 254 nm % B App ID min Sample Pre-treatment To.5 g (+/-.5 g) Baby Wash and Shampoo add 1.25 ml acetone and.6 ml 1 % NaCl solution. Vortex for 3 seconds. Wait for the emulsion to dissipate (about 3 seconds) and centrifuge for 1 minutes at 3, rpm. Collect the supernatant. Novum SLE Procedure Load the supernatant from the pre-treatment step onto a Novum SLE 12 cc tube. Apply a short and gentle pulse of vacuum (~5-1 seconds at 5 Hg) until the sample has completely entered the media. Wait for 5 minutes. Dispense 1 ml Ethyl acetate onto the Novum SLE sorbent and collect solvent under gravity (~1 minutes). Apply vacuum at 5-1 Hg to complete the extraction. Analyte % Absolute Recovery %CV (N=8) Methyl paraben Ethyl paraben Propyl paraben Butyl paraben Dry down and reconstitute in.5 ml Methanol. Centrifuge for 1 minutes at 6, rpm. Collect supernatant and inject onto HPLC. Phenomenex l WEB: 9

10 A Variety of Formats to Fit Your Sample and Throughput Requirements Tubes Process samples as small as 1 μl or as large as 1 ml using Novum SLE tubes. Ideal for all types of applications including Bioanalytical, Food Safety, and Environmental. Novum SLE Tubes 1 cc 3 cc 6 cc 12 cc Maximum Sample Volume (before dilution) 1 µl 2 µl 5 µl 1 ml Recommended Elution Volume 1.2 ml 1.8 ml 5 ml 1 ml 96-Well Plates Process 96 samples at once in an easily automatable 96-well plate. Perfect for high throughput applications. Novum SLE 96-Well Plates MINI MAX Maximum Sample Volume (before dilution) 15 µl 2 µl Recommended Elution Volume 1 ml 1.8 ml 1

11 Novum Simplified Liquid Extraction (SLE) Products 96-Well Plates Part No. Description Unit Price 8E-S138-FGA Novum SLE MINI 96-Well Plate 1/pk 8E-S138-5GA Novum SLE MAX 96-Well Plate 1/pk 96-Well Plate Accessories Part No. Description Unit Price Collection Plates (deep well, polypropylene) AH Well Collection Plate, 35 µl/well 5/pk AH Well Collection Plate, 1 ml/well 5/pk AH Well Collection Plate, 2 ml/well 5/pk AH Well Collection Plate, 2 ml/well Square/Round-Conical 5/pk AH Well Collection Plate, 2 ml/well Round/Round, 8 mm 5/pk AH Well Collection Plate, 1 ml/well Round, 7 mm 5/pk Sealing Mats AH-8597 Sealing Mats, Pierceable, 96-Square Well, Silicone 5/pk AH-8598 Sealing Mats, Pre-Slit, 96-Square Well, Silicone 5/pk AH-8631 Sealing Mats, Pierceable, 96-Round Well 7 mm, Silicone 5/pk AH-8632 Sealing Mats, Pre-Slit, 96-Round Well 7 mm, Silicone 5/pk AH-8633 Sealing Mats, Pierceable, 96-Round Well 8 mm, Silicone 5/pk AH-8634 Sealing Mats, Pre-Slit, 96-Round Well 8 mm, Silicone 5/pk AH-7362 Sealing Tape Pad 1/pk Vacuum Manifold AH Well Plate Manifold, Universal with Vacuum Gauge ea Tubes Part No. Description Unit Price 8B-S138-FAK Novum SLE 1 cc tubes 1/pk 8B-S138-5BJ Novum SLE 3 cc tubes 5/pk 8B-S138-JCH Novum SLE 6 cc tubes 3/pk 8B-S138-KDG Novum SLE 12 cc tubes 2/pk Tube Accessories Vacuum Manifolds Part No. Description Unit Price AH Position Vacuum Manifold Set ea AH Position Vacuum Manifold Set ea If Novum 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. Terms and Conditions Subject to Phenomenex Standard Terms and Conditions, which may be viewed at Trademarks Kinetex is a registered trademark, Novum and Synergi are trademarks of Phenomenex. Freedom EVO is a registered trademark of Tecan Group Ltd. API 5 is a trademark of AB SCIEX Pte. Ltd. AB SCIEX is being used under license. Novum is patent pending. 215 Phenomenex, Inc. All rights reserved. Phenomenex l WEB: 11

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