Agilent Captiva EMR Extractor Method Guide PDF

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Summary of Content for Agilent Captiva EMR Extractor Method Guide PDF

Agilent Captiva EMRLipid Method Guide General instructions for 96-well plate and 1 mL cartridge formats: Agilent Captiva EMRLipid cartridges and 96-well plates allow streamlined in-well protein precipitation, filtration, and cleanup for lipid-containing samples. The improved filter design gives easy flow with vacuum or positive pressure, and allows for protein precipitation without clogging during elution. The novel EMRLipid sorbent chemistry provides highly selective and efficient lipid/matrix removal without impacting analyte recovery. Effective matrix removal assures minimal ion suppression or enhancement on target analytes, which significantly improves method reliability and ruggedness.

The 96-well plates are ideal for high-throughput workflows, while 1 mL cartridges can accommodate small batch needs.

1. Add biological sample and crash solvent

Solvent Sample

2. Mix to precipitate protein 3. Filter

Salts Proteins Lipids Analyte

Option 1 Passive mixing: let stand for 5 minutes

Option 2 Active in-well mixing: vortex or mix with wide-bore pipette tips

It is highly recommended to add sample rst and then crash solvent, to achieve better sample homogeneity during sample and solvent addition.

Operating instructions and tips for Agilent Captiva EMRLipid 96-Well Plate and 1 mL Cartridge Products

User Tips Protein precipitation workflow

Product configuration 96-well plate or 1 mL cartridge

Sample size Between 20200 L

Sample treatment Crash solvent ratio: between 3:1 and 5:1 ACN with 1 % formic acid to sample. If total volume is less than 500 L, add additional 4:1 ACN:H2O to reach a minimum volume of 500 L. ACN is preferable to MeOH to maximize protein precipitation and avoid gelation.

Recommended sample addition order

1) Sample 2) Crash solvent

Mixing Option 1: Passive mixing. Let stand for 5 minutes to allow for complete protein precipitation to occur. Option 2: Active in-well mixing. For 96-well plates containing >500 L of total volume and 1 mL cartridges regardless of volume, pipette mixing using wide bore pipette tips (perform 3 to 5 aspiration/dispense cycles) is recommended. For 96-well plates with <500 L of total volume, cap and vortex at 1,350 rpm. Option 3: Protein precipitation and mixing can be performed in a separate tube, centrifuged, and subsequently transferred to the Captiva EMRLipid well/cartridge.

Pass-through filtration and cleanup

Vacuum between 25 in Hg initiates flow. Positive pressure (34 psi) is also acceptable. For optimal lipid removal, a controlled flow rate of one drop every 35 seconds is highly recommended. After elution, apply higher vacuum or positive pressure for 12 minutes to ensure maximum sample recovery. Flow rate is dependent on sample type, crash solvent, and mixing. An alternative approach to vacuum and positive pressure is centrifugation. For 96-well plates, 500800 rpm for a minimum of 10 minutes is recommended, followed by 12 minutes at 2,5003,000 rpm to drain the cartridge. Centrifugation speed and time are dependent on the sample volume and matrix.

For more detailed protocol recommendations, please refer to Agilent Application Note 5991-9222EN.

Agilent Captiva EMRLipid ordering information

Part number Description Quantity

5190-1000 Agilent Captiva EMRLipid 96-well plate

1 plate

5190-1001 Agilent Captiva EMRLipid 96-well plate

5 plates

5190-1002 Agilent Captiva EMRLipid 1 mL cartridge

100/pk

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