[ Care and Use ManUal ]

1Sirocco Protein Precipitation Plate

Sirocco Protein PreciPitation Plate

The Sirocco™ plate was designed to improve the automation and through-

put of protein precipitation in drug metabolism/ADME/toxicology labs.

The Sirocco Protein Precipitation Plate is a unique combination of novel

96-well plate tip design with proprietary new filter membranes.

A straightforward 7-step procedure designed to enable precipitate free

sample filtrates suitable for high-throughput LC/MS analysis.

Step 1 – Preparation

Place the Sirocco Plate on a collection plate for processing. This protects

the valve tips during the processing steps.

Sirocco ProteinPrecipitation Plate

96-Well Collection Plate

Step 2 – Add Crash Solvent

Add precipitation solvent to all wells. Use a ratio of at least 3:1 solvent

to sample. Acetonitrile is the recommended precipitation solvent. The

Sirocco plate is designed to prevent solvent from leaking through the

plate at this step.

Step 3 – Load Sample

Add 20-200 µL of sample (plasma, urine, etc.) rapidly into the wells. Rapid

delivery of sample into the crash solvent creates mixing and initiates

precipitation process.

[ Care and Use ManUal ]

2Sirocco Protein Precipitation Plate

Step 4 – Seal

Apply vented cap mat, using a cap roller to ensure secure, uniform sealing.

This will prevent splashing and cross-talk during mixing.

Cap Mat

Step 5 – Mix

Shake (mix) the loaded Sirocco plate/collection plate assembly on a vortexer

(e.g., Multi-Tube Vortexer from VWR) at medium setting (e.g., setting 5 on a

1-10 scale) for 0.5 to 1.0 minute.

Note: There is no need to remove the vented cap mats or valve tips from the

Sirocco Plate before filtration. This eliminates the risk of cross-talk between

wells. The valves and vented cap mat are designed to open under vacuum

to allow controlled flow during filtration.

Note: Excessive mixing at too high a setting (e.g., setting 8 on a 1-10

scale) could cause cross-talk. Under mixing (e.g., setting 2 on a 1-10 scale)

will cause clogging or a cloudy filtrate.

Step 6 – Filter

After mixing, place the Sirocco plate on a Vacuum Manifold (e.g. Waters

vacuum manifold P/N 186001831), and apply 8-5” Hg (271-508 mbar)

of vacuum for up to three minutes. A higher vacuum setting can be used if

flow does not occur at 10” Hg.

Note: The Sirocco proprietary valve tip design prevents liquid flow through

the device until a minimum 5” Hg (169 mbar) of vacuum is applied.

Note: The Sirocco Plate can be filtered by using centrifiguation (@ 2,000 x g

for 1 min.) in place of vacuum device.

Step 7– Analysis

Discard the Sirocco Plate and remove the collection plate containing clear

filtrate from the vacuum device, seal and transfer to the LC/MS autosam-

pler or store for later analysis.

Note: There is no need to remove cap mats or valve tips from Sirocco Plate.

The Sirocco Plate is discarded after filtration.

Xevo™ TQ MS ACQUITY® SM BSM

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3

Ordering inFOrMAtiOn

description Volume Qty/Pkg Part number

Sirocco Protein Precipitation Plate (includes 1 plate and 1 cap mat)

– 1/pkg 186003873

Sirocco Protein Precipitation Plate (includes 5 plates and 5 cap mats)

– 5/pkg 186002448

Austria and European Export (Central South Eastern Europe, CIS and Middle East)

43 1 877 18 07

Australia 61 2 9933 1777

Belgium 32 2 726 1000

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Canada 1 800 252 4752 x2205

China 86 21 6879 5888

CIS/Russia

+497 727 4490/290 9737

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420 2 617 1 1384

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Finland 09 5659 6288

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The Netherlands

31 76 508 7200

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Switzerland 41 56 676 70 00

Taiwan 886 2 2543 1898

United Kingdom

44 208 238 6100

All other countries: Waters Corporation U.S.A. 1 508 478 2000

1 800 252 4752

www.waters.com

Sales Offices

Waters Corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com

© 2010 Waters Corporation. Waters, The Science of What’s Pos-sible, Xevo and ACQUITY are trademarks of Waters Corporation. Pall, , and Sirocco are trademarks of Pall Corporation. VWR is a trademark of VWR International.

August 2010 720003377EN Rev B KK-PDF