A simple GC method for determination of cryoprotector diols 1,4-butanediol or 2,3-butanediol in isolated rat hepatocytes.
We devised a simple method for determining the cryoprotectant agents 1,4-butanediol or 2,3-butanediol in isolated rat hepatocytes. After extraction of of hepatocytes with water (containing internal standard - ethylene glycol 1.25 mg/mL) the diol content was analyzed by gas chromatography. The method shows a linear response in the range 0.125 to 2.50 mg/mL for 1,4-butanediol and 0.25 to 3.75 mg/mL for 2,3-butanediol. The accuracy and precision of the method were evaluated and the coefficients of variation were found to be within = 6.0 %. The recoveries from hepatocyte samples containing 0.50, 1.00 and 2.00 mg/mL were 91.0 to 108 % for 1,4-butanediol and 80.6 to 100.3 % for 2,3-butanediol, respectively. This method allowed the determination of the intracellular concentration of diols in hepatocytes preserved for up to 120 hours at - 4 (C in UW solution + 8 % w/v 1,4-butanediol (or 2,3-butanediol).
Almada
Luciana
L
Farmacología, Depto Cs. Fisiológicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, Rosario (S2002LRK), Argentina.
Guibert
Edgardo E
EE
Rodriguez
Joaquin V
JV
D000818:Animals; D002072:Butylene Glycols / Q000032:analysis*; D002849:Chromatography, Gas / Q000379:methods*; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000032:analysis*; D022781:Hepatocytes / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar
D002072:Butylene Glycols; D003451:Cryoprotective Agents; C026978:2,3-butylene glycol; C039681:1,4-butanediol