A simple, differential extraction method for the simultaneous direct radioimmunoassay of androgens and androgen glucuronides in human serum.
A method is described for the differential extraction of unconjugated androgens (testosterone and dihydrotestosterone) and glucuronidated androgens (androstane-3 alpha,17 beta-diol glucuronide and androsterone glucuronide) from human serum using solid-phase, gravity-flow extraction columns. In this method, 100-microL aliquots of serum are loaded onto the normal-phase columns, unconjugated androgens are eluted with ethyl ether, and glucuronides are eluted with ethyl ether containing 2% acetic acid. Glucuronide eluates are washed with 1% aqueous acetic acid to remove cross-reacting steroid sulfates. Assays of sera for the four steroids were performed using standard radioimmunoassay methodology, except for androsterone glucuronide. This steroid was assayed with a novel radioimmunoassay method that employees a tritiated, unconjugated androsterone tracer and an anti-dehydroepiandrosterone sulfate antiserum. The new method is well suited for the assay of conjugated and unconjugated steroids in large numbers of specimens, particularly where the sample volume is limited.
Brind
J
J
Orentreich Foundation for the Advancement of Science, Inc., Cold Spring, New York 10516, USA.
Borofsky
N
N
Chervinsky
K
K
Vogelman
J H
JH
Orentreich
N
N
D000328:Adult; D000368:Aged; D000728:Androgens / Q000097:blood* / Q000737:chemistry; D015113:Androstane-3,17-diol / Q000031:analogs & derivatives / Q000097:blood / Q000737:chemistry; D000738:Androsterone / Q000031:analogs & derivatives / Q000097:blood / Q000737:chemistry; D013196:Dihydrotestosterone / Q000097:blood / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D011863:Radioimmunoassay / Q000379:methods*; D012016:Reference Values; D013739:Testosterone / Q000097:blood / Q000737:chemistry; D014316:Tritium
D000728:Androgens; D013196:Dihydrotestosterone; D014316:Tritium; C027927:androsterone glucuronide; D015113:Androstane-3,17-diol; C028895:androstane-3,17-diol glucuronide; D013739:Testosterone; D000738:Androsterone