Sample Preparation Products. TELOS® PPT. Protein Precipitation Columns &
Plates. Introduction. Protein precipitation is a commonly used sample preparation
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How to use Guide
Sample Preparation Products
TELOS PPT ®
Protein Precipitation Columns & Plates Introduction Protein precipitation is a commonly used sample preparation technique prior to analysing drugs and drug metabolites in biological fluid samples, e.g. plasma. With acetonitrile as the preferred precipitation (crash) solvent, the subsequent supernatant is transferred to a suitable vial or plate. TELOS® PPT Protein Precipitation Columns and Plates use filtration to minimise the number of liquid transfer steps and maximise the removal of proteins and large molecular weight peptides.
TELOS PPT Protein Precipitation Columns and Plates TELOS PPT Protein Precipitation Columns and Plates use a proprietary frit to maximise precipitation, so minimising the levels of precipitated and suspended materials in the final filtrate. Acetonitrile is added to each column or well, presenting a “pool” of solvent for the plasma sample. Adding the sample to the acetonitrile with force encourages efficient mixing, ensuring the maximum levels of peptides and proteins are precipitated. The application of a vacuum allows the filtrate to pass into the collection plate or vial, leaving colloidal materials and precipitate in the filterplate. See Figure 1.
Add Solvent
Dispense Sample
Apply Vacuum
(with force)
Wait for 3 - 5 minutes
Precipitation Occurs “Pool” of solvent
No Solvent Dripping Dual Fritted Filtration
Precipitate is
Figure 1. Protein Precipitation Procedure using TELOS PPT Columns and Plates
Sample Preparation Method Guidelines This section comprises a general method that can be used as a starting point for new protein precipitation methods, followed by additional guidelines for particular steps of the procedure. General Procedure for use of TELOS PPT Columns Step 1: Place the required number of TELOS PPT Columns on a sample processing manifold. Ensure the appropriate collection vial is placed beneath each column Step 2: Add acetonitrile (400μl) to each column – no dripping will occur Step 3: Add plasma (100μl) with force to the centre of each column; again, no dripping will occur. Wait for 3 – 5 minutes Step 4: Apply vacuum (~-20”Hg) and collect the filtrate (3 minutes is normally sufficient)
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Collect Filtrate
Removed
How to use Guide
TELOS PPT
Protein Precipitation Columns & Plates General Procedure for use of TELOS PPT Plates Step 1: Place the TELOS PPT Plate onto a 96-well plate sample processing manifold. Ensure the appropriate collection plate is placed beneath the plate Step 2: Add acetonitrile (400μl) to each well – no dripping will occur Step 3: Add plasma (100μl) with force to the centre of each well; again, no dripping will occur. Wait for 3 – 5 minutes Step 4: Apply vacuum (~-20”Hg) and collect the filtrate (3 minutes is normally sufficient)
Optimum Acetonitrile/Plasma Ratio The optimum acetonitrile/plasma ratio is 4:1 v/v. This combination provides maximum precipitation of plasma peptides and proteins. The acetonitrile/plasma ratio can be increased; however, this could lead to incomplete precipitation.
Minimum Plasma Sample Volume The minimum plasma sample volume which can be processed (using the optimum 4:1 v/v ratio) is ~15μl. This requires 60μl of acetonitrile, the minimum practical volume to provide a “pool” at the bottom of each column or well.
Maximum Plasma Sample Volume Columns
The maximum plasma volume which can be processed using TELOS PPT Columns is 200μl using the 4:1 v/v ratio.
Plates
The maximum plasma volume which can be processed using TELOS PPT Plates is 400μl using the 4:1 v/v ratio.
Alternative Sample Types Serum or whole blood samples may be processed using TELOS PPT Columns and Plates. For such samples, the acetonitrile/sample ratio will require optimisation.
Alternative Precipitation Solvents Aqueous precipitation solvents, e.g. trichloroacetic acid, can be used. N.B. Methanol is not suitable for use with TELOS PPT Columns and Plates.
Ordering Information Part Number
Description
960-0001-096P
TELOS PPT Protein Precipitation Plate
Pack Size
960-0001-001T
TELOS PPT Protein Precipitation Columns, 1ml
EACH 100
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