Metabolomics Facilitated by High-Throughput Sonication

Metabolomics is the research field that employs high-throughput techniques to characterize and quantify small molecules in various biofluids, such as urine, serum, plasma, feces, and saliva, unveiling unique metabolic signatures. The high-throughput sonicator UIP400MTP, designed for multi-well plates, is a powerful tool for lysing cells in these samples, facilitating the release of metabolites for downstream analysis. This precise and reliable approach is essential to metabolomics, where accurate study design, meticulous sample preparation, optimized platform setup, comprehensive data analysis, and meaningful biological interpretation are paramount. Metabolomics employs both targeted approaches, which focus on specific classes of molecules like carbohydrates, lipids, and amino acids, and non-targeted techniques, which provide a broad overview of the metabolic profiles within a sample, revealing the intricate complexity of metabolic signaling.

High-Throughput Sample Preparation with the 96-Well Plate Sonicator UIP400MTP

Using the non-contact sonicator UIP400MTP in combination with 96-well plates is an efficient method for high-throughput homogenization and metabolite extraction. This setup avoids direct contact with the sample, reducing the risk of cross-contamination while processing multiple samples simultaneously.

Step-by-Step Protocol for Homogenization with 96-Well Sonicator UIP400MTP

1. Preparation of Metabolomic Samples
   Sample Placement: Dispense each sample (e.g., feces, gut tissue, or luminal contents) into individual wells of a 96-well plate or microplate.
   Solvent: Organic solvents, such as methanol, ethanol, acetonitrile, chloroform, and their (aqueous) mixtures, are essential for metabolite extraction, aiding in the release of intracellular metabolites from the cellular matrix. Use 100–300 µL of solvent per well, ensuring enough volume to submerge the sample fully.
   Internal Standards: Optionally, add internal standards to each well for normalization during downstream metabolomic analysis.
2. Coupling Medium Setup
   Water Level: Place the 96well plate on the sonication platform filled with a water (the coupling medium) as described in the manual.
3. UIP400MTP Parameter Configuration
   Power Settings: Adjust ultrasonic amplitude to 50–80% of the maximum power for efficient homogenization.
   Pulse Mode: Set the sonication cycle to alternate pulses (e.g., 30 seconds ON, 30 seconds OFF) to allow for cooling and prevent sample overheating.
   Total time: 4–10 minutes, depending on sample complexity and toughness.
   Temperature Control: Use a lab chiller to maintain the water at 4–8°C during sonication to protect heat-sensitive metabolites.
4. Sonication Process
   Start the UIP400MTP for sonication cycles.
   Monitor the process via the UIP400MTP interface.
   The cavitation effect will homogenize samples and lyse cells within the wells, releasing intracellular metabolites into the solvent.
5. PostSonication
   Centrifugation: After sonication, transfer the plate contents to collection tubes if needed. Centrifuge at 4,000–6,000 × g for 10 minutes to pellet insoluble debris and cellular material.
   Supernatant Collection: Carefully transfer the supernatant from each well into new tubes or plates for analysis.
   Optional Drying: Dry supernatants using a vacuum concentrator (e.g., SpeedVac) if concentration is required, then reconstitute in a small volume of the appropriate solvent.

Take advantage of the high-throughput processing capacity of the UIP400MTP multi-well plate sonicator and facilitate your metabolomic research and analysis!

Principle of Non-Contact Sonication with UIP400MTP

The UIP400MTP applies ultrasonic waves to the wells of a 96-well plate indirectly via a coupling medium (e.g., water). These waves create cavitation, which disrupts cells and releases metabolites without physical contact between the sonicator probe and the sample.

Design, Manufacturing and Consulting – Quality Made in Germany

Hielscher ultrasonicators are well-known for their highest quality and design standards. Robustness and easy operation allow the smooth integration of our ultrasonicators into industrial facilities. Rough conditions and demanding environments are easily handled by Hielscher ultrasonicators.

Hielscher Ultrasonics is an ISO certified company and put special emphasis on high-performance ultrasonicators featuring state-of-the-art technology and user-friendliness. Of course, Hielscher ultrasonicators are CE compliant and meet the requirements of UL, CSA and RoHs.