Agilent Bond Elut 1 mL cartridge Extractor Method Guide PDF

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Summary of Content for Agilent Bond Elut 1 mL cartridge Extractor Method Guide PDF

Agilent Bond Elut Lipid Extraction

Method guide for 1 mL cartridge

General instructions Agilent Bond Elut Lipid Extraction cartridges allow the extraction and isolation of lipids from biological samples such as plasma, cell cultures, and tissue samples. The unique EMRLipid sorbent chemistry provides effective and selective retention of lipid compounds on the sorbent. After a washing step, the trapped lipid compounds are eluted with solvent. The 1 mL SPE cartridge format simplifies the extraction process compared to traditional liquidliquid extraction techniques used for lipid analysis. The workflow provides significant time savings with equivalent extraction efficiency and improved method reproducibility. Bond Elut Lipid Extraction 1 mL cartridges are suitable for small batch sample preparation.

For more information, visit: www.agilent.com/chem/BondElutLE

Biological sample

Add crashing solvent at

a ratio of 9:1 for protein precipitation

Transfer entire homogenate to

a Bond Elut Lipid Extraction

cartridge

Discard the eluent

Wash

Discard the eluent

Elute

Protein precipitation Sample loading Sample washing Lipids elution

Collect the eluent

1. Crashing solvent: ACN with 1 to 5% MeOH. A small percentage of MeOH helps to generate finer protein precipitates, which allows for easy pipette transfer of the homogenate.

2. Washing solvent: a mixture of ACN with 10 to 20% of water. 3. Elution solvent: a solution of MeOH with dichloromethane (DCM), chloroform, or

1-chlorobutane. A minimum of 50% MeOH is important for the release of lipids. Agilent recommends DCM/MeOH (v/v 1:2) or chloroform/MeOH (v/v 1:1).

www.agilent.com/chem/BondElutLE

For Research Use Only. Not for use in diagnostic procedures.

This information is subject to change without notice.

Agilent Technologies, Inc. 2020 Published in the USA, February 3, 2020 5994-1627EN DE.3386458333

A recommended Bond Elut Lipid Extraction protocol for plasma extraction 1. Add 100 L of plasma to a 1.5 mL Eppendorf tube.

2. Add 900 L of ACN with 1 to 5% MeOH. Vortex for 30 seconds, and sonicate on ice for 10 minutes.

3. Transfer the entire homogenate to a Bond Elut Lipid Extraction 1 mL cartridge. Apply low vacuum or positive pressure for steady elution. Discard the eluent.

4. Add 2 1 mL of ACN/water (v/v 9:1) for wash. Rinse the original sample tube with wash, then transfer it to the cartridge for flowthrough. Apply vacuum or positive pressure as needed for a smooth flow. Discard the eluent.

5. After the wash has eluted, apply higher vacuum or pressure to dry the cartridge. Place a glass collection tube beneath the cartridge.

6. Add 2 1 mL of DCM/MeOH (v/v, 1:2) for elution, and collect the eluent.

7. Dry the eluent with N2 at 30 C.

8. Reconstitute into 100 L of n-BuOH/MeOH (v/v, 1:1), vortex for two minutes, and sonicate for 10 minutes.

9. Transfer to a 2 mL vial with an insert, and cap for MS analysis or store in a 20 C freezer.

Product use tips and tricks 1. Maintain a slow flow rate of approximately 3 to 5 seconds per drop.

2. Use a ratio of 9:1 crashing solvent/plasma for protein precipitation to improve lipid extraction efficiency. The EMRLipid sorbent requires a minimum of 10% aqueous in the sample mixture for loading.

3. It is important to transfer the entire homogenate, as lipids can be trapped in the protein precipitate. Do not centrifuge before transferring.

4. Wide bore pipette tips are recommended for transfer.

Bond Elut Lipid Extraction ordering information

Description Quantity Part Number

Agilent Bond Elut Lipid Extraction, 1 mL ca

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