The effect of formalin and the duration of acid hydrolysis on free and conjugated urinary 17-ketosteroids.

The recovery of five free ketosteroids, three glucuronides, and six sulfates, added to water or urine and subjected to acid hydrolysis for various lengths of time, was investigated. Dehydroepiandrosterone (DHEA) in all its forms was most susceptible to destruction. The behavior of glucuronides and sulfates differed, and no single set of conditions provided equal recovery for all conjugates. A compromise time of 30 mm. of hydrolysis appeared to be optimal. The addition of formalin diminished nonspecific chromogens in a variable manner, but also accelerated destruction of DHEA and decreased the recovery of ketosteroid sulfates.