Simultaneous ultramicroanalysis of both 17-keto-and 17beta-hydroxy androgens in biological fluids.
Sensitive methods for quantifying androgens were lacking. Therefore, a relatively simple procedure for separating steroids was combined with highly specific assay methods so that eight androgens could be measured with high accuracy, precision and sensitivity. Semi-automated separations on Sephadex LH-20 columns used heptane:methylene chloride:ethanol:water (50:50:1:0.12) and a flow rate of 17.0 min/ml. The six peaks eluted contained androstenedine; androsterone, epiandrosterone and dihydrotestosterone; testosterone and dehydroepiandrosterone; 3alpha-androstanediol; 3beta-androstanediol; and androstenediol. Androstenedione, dehydroepiandrosterone and androstenediol were quantified using specific antisera (sensitivity less than or equal to 75 pg). Testosterone and dihydrotestosterone were measured by competitive protein-binding assays using rabbit TeBG (sensitivity less than or equal to 150 pg). 3alpha- and 3beta-androstanediol were similarly assayed using human TeBG (sensitivity approximately 150 pg). Androsterone was reduced with NaBH4 and the resulting 3alpha-androstanediol was assayed using human TeBG (sensitivity approximately 200 pg). Inter- and intra-assay variations were less than 10% for radioimmunoassays and less than 16% for competitive protein-binding assays over the entire dose response curve.
D000728:Androgens / Q000032:analysis* / Q000097:blood; D015113:Androstane-3,17-diol / Q000032:analysis; D000735:Androstenedione / Q000032:analysis; D000738:Androsterone / Q000032:analysis; D000818:Animals; D001826:Body Fluids / Q000032:analysis*; D001894:Borohydrides / Q000378:metabolism; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods*; D003429:Cross Reactions; D003687:Dehydroepiandrosterone / Q000032:analysis; D004822:Epididymis / Q000098:blood supply; D005069:Evaluation Studies as Topic; D008297:Male; D010084:Oxidation-Reduction; D011863:Radioimmunoassay / Q000379:methods; D011869:Radioligand Assay / Q000379:methods; D012152:Rete Testis / Q000378:metabolism; D013237:Stereoisomerism; D013739:Testosterone / Q000032:analysis
D000728:Androgens; D001894:Borohydrides; D015113:Androstane-3,17-diol; D013739:Testosterone; D000735:Androstenedione; D003687:Dehydroepiandrosterone; D000738:Androsterone