Bead-beating tissue homogenisation technology
A powerful and high throughput sample prep tool for quantitation of analyte in bone, skin & lymph node
By Bhavesh Patel, Director – Head DMPK-Bioanalysis, Eurofins Advinus Limited
Estimation of drug concentrations in tough tissues such as skin, lymph nodes, ocular tissues and bones is critical for many preclinical pharmacokinetic studies. For example, drug concentrations and exposure in bones is determined during development of drugs for osteoporosis, rheumatoid arthritis and bone cancer. Analyte extraction from these tough tissues, however, is plagued with inherent challenges. Traditionally, such extraction is accomplished using mechanical or manual homogenisers, mortar and pestle, and sonicators. Such sample preparation techniques require the tough tissues to be cryo-frozen to make them brittle as a first step followed by grinding. This procedure is prone to sample cross contamination. These procedures are time consuming and low throughput. Also, the mechanical processes involved result in elevated temperatures, which may be undesirable for the sample integrity.
The application of the bead-beater-blender resolves many of these challenges. The bead beater methodology involves grinding the tough biological samples with small beads in micro-centrifuge tubes for a brief period of about 15 minutes. Beads of different materials are available (stainless steel, zirconium, ceramic, etc.) and can be optimised based on the application. The device is cooled at 4 °C and insulated for noise. This simple approach enables fast and complete homogenisation of tissues as well as cultured cells. Simultaneously, 24 samples (0.3 g each) can be processed in individual microfuge tubes.
Using this platform, Eurofins Advinus has successfully developed robust sample preparation methods for these tough tissues. The newly procured “Bullet Blender Gold” agitates the sample tubes vigorously and provides uniform and reproducible homogenisation while preserving sample integrity. The platform has been used to efficiently extract drugs from skin, lymph nodes and tibia. All consumables involved in sample preparation are disposable; thus, the problems of cross contamination and sample loss are resolved. The effectiveness of this sample preparation procedure improves the quality and throughput of the results generated.
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