What is the standard operating procedure for the Sirocco Protein Precipitation Plate (186002448 or 186003873)? - WKB119433

ENVIRONMENT

• Sirocco

ANSWER

The standard operating procedure for the Sirocco Protein Precipitation Plate is as follows:

Step 1 - Preparation

Set the Sirocco plate on the collection plate. This protects the tip of the valve during processing.

Step 2 - Addition of organic solvent for precipitation (acetonitrile)

Add precipitation solvent to all wells. Use a solvent ratio of at least 3:1 to the sample. The recommended precipitation solvent is acetonitrile. The Sirocco plate is designed to prevent solvent from leaking out of the plate during this step.

Step 3 - Load sample

Quickly add 20-200 µL of sample (e.g., plasma, urine) to the wells. Add the sample quickly to the organic solvent for precipitation, and mix to start the precipitation process.

Step 4 - Seal

Use the cap roller to set the venting cap mat for reliable and uniform sealing. This will prevent droplets and crosstalk during mixing.

Step 5 Mix

Shake (mix) the loaded Sirocco plate / collection plate assembly with a vortex (such as VWR's Multi-Tube Vortexer) using a medium setting (such as 5 on a scale of 1-10) for 0.5-1.0 minutes.

Note: It is not necessary to remove the venting cap mat or valve tip from the Sirocco plate before filtering. This eliminates the risk of crosstalk between wells. The valve and venting cap mats are designed to open under vacuum, allowing you to control the flow during filtration. Note: Excessive mixing with a high setting(e.g., setting 8 on a scale of 1-10) can cause crosstalk. During mixing (for example, setting 2 on a scale of 1 to 10), clogging and turbidity of the filtrate can occur.

Step 6 - Filter

After mixing, place the Sirocco plate on a vacuum manifold (e.g., Waters vacuum manifold 186001831) and apply a vacuum of 8-5 in Hg (271-508 mbar) for up to 3 minutes. A higher vacuum setting is possible if there is no flow at 10 in. Hg. Note: Sirocco's unique valve tip design prevents liquid from flowing through the device until a vacuum of at least 5 in. Hg (169 mbar) is applied. Note: Scirocco plates can be filtered using a centrifuge (@ 2,000 × g for 1 minute) instead of a vacuum system.

Step 7 - Analysis

Remove the Sirocco plate, remove the collection plate containing the clear filtrate from the vacuum system, seal and transfer to an LC/MS autosampler, or store for later analysis. Note: There is no need to remove the cap mat or valve tip from the Sirocco plate. Sirocco plates are discarded after filtration.

ADDITIONAL INFORMATION

Sirocco Protein Precipitation Plate 720003377EN Care & Use
https://www.waters.com/waters/suppor...1442&type=USRM