Quantitation of dimethyl sulfoxide in solutions and tissues by high-performance liquid chromatography.
We have developed a rapid and simple method to determine the level of dimethyl sulfoxide (Me2SO) in both solutions and tissue samples. For analysis of Me2SO in a cryopreservation medium, the solution is simply diluted in 10% (vol/vol) methanol and centrifuged. Then an aliquot of the supernatant is assayed by high-performance liquid chromatography. For tissue samples, the wet weight is measured and the intact sample is extracted with 10% (vol/vol) methanol (e.g., 10 ml/g wet wt) in a sealed vial. The extract is then diluted and centrifuged, and an aliquot of the supernatant is assayed. The dry weight of the tissue is measured after the methanol-extracted sample is placed into either for 2 h and air-dried overnight. The water content of the tissue is calculated as the difference between the wet and the dry weights. The concentration of Me2SO in the aqueous compartment of the tissue can then be calculated by taking into account the concentration of Me2SO in the extract and the dilution factor, based on the tissue water volume and the volume of methanol used to extract the Me2SO. The calculated values for porcine myocardium samples correlated 1:1 with the actual Me2SO concentrations in the solutions in which the tissue samples were equilibrated. Finally, we present results documenting the usefulness of this assay by following the time course of Me2SO penetration into core versus peripheral regions of 1-cm3 samples of porcine myocardium.
Carpenter
J F
JF
CryoLife, Inc., Marietta, Georgia 30067.
Dawson
P E
PE
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003451:Cryoprotective Agents / Q000032:analysis; D004121:Dimethyl Sulfoxide / Q000032:analysis*; D009206:Myocardium / Q000737:chemistry; D012996:Solutions; D013552:Swine; D014018:Tissue Distribution
D003451:Cryoprotective Agents; D012996:Solutions; D004121:Dimethyl Sulfoxide