PROTOCOLS

Steroid Solid Extraction Protocol

We recommend an organic phase extraction to extract steroids from non-liquid matrices such as dried solids or other organic matter. We use ethanol or ethyl acetate as safer alternatives to diethyl ether or methylene chloride. We recommend centrifugal vacuum devices (i.e., a SpeedVacTM) to remove the solvent completely and safely after extraction. Alternatively, the organic layer can be separated and stored for later use. We also recommend using ethanol to completely solubilize the dried extracted steroid since steroids typically have limited aqueous solubility.

Steroid Liquid Extraction Protocol

Efficient extraction of samples might be a necessity for an accurate determination of total steroid concentrations in serum, plasma, saliva and other liquid samples. Extraction may also be necessary to concentrate the sample to within the assay’s measurement range. Extraction eliminates potential interfering substances, such as bulk proteins and lipids.We would recommend using at least 1-2 mL of serum or plasma for the steroid measurement If the assay requires serum or plasma samples to be concentrated. As with any purification technique, recovery of the desired substance is likely to be incomplete. Therefore, both optimization and quantification of the extraction procedure are recommended for accurate determinations.