The Effect of Formalin and the Duration of Acid Hydrolysis on Free and Conjugated

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. [1OTAL URINARY l7-lETos’rERolDs are still widely measured, in spite of serious problems of specificity, accuracy, and interpretation (1-5). The large and variable proportion of Ilonketosteroid chromogemis measured by routine proce(lum’es has beemi repeatedly documented (1,2). Numerous attempts have been made to nlimiimize this problem while retaining tile si)eed ahl(l simplicity required for a routine clinical method. From time to time foi’maliii has been mentioned as a chromogen-destroying agent (6), although its action amid specificity have not l)een documented. In addition to these prohienls, studies of tile recovery of added steroids have suffered from tile fact tilat omilv free compounds were available, rather than tile conjugates, w’hich are present iii urine. The assumption that acid hydrolysis acts in identicalfashion on free steroids, sulfates, and giucuromiides requimes proof. Since there is still need-in spite of the availability of highly specific 1)ut rather cumbersonle methods for measuring individual steroids (7-9)-of a rapid clinical inetilod fo