Development of highly sensitive quantification method for testosterone and dihydrotestosterone in human serum and prostate tissue by liquid chromatography-electrospray ionization tandem mass spectrometry.
We developed highly sensitive detection of testosterone (T) and 5alpha-dihydrotestosterone (DHT) by liquid chromatography-electrospray ionization tandem mass spectrometry using high proton affinitive derivatization of 17beta-hydroxyl group of T and DHT with picolinic acid, mobile phase consisting of MeCN-MeOH-H(2)O-formic acid and conventional octadecylsilica (ODS) column. Purification of the derivatives was carried out using solid-phase extraction with ODS cartridge. By this method, T and DHT were determined simultaneously with limits of quantification (LOQs) of 1 pg/0.2 ml in serum, and T and DHT with LOQs of 0.5 pg and 1 pg/3mg in prostate tissue, respectively, under acceptable assay performance (intra-assay and inter-assay accuracy and precision). The present method provides reliable and reproducible results for quantification of T and DHT in small volumes of serum and prostate samples for diagnosis in prostatic disorders and male climacteric.
Yamashita
Kouwa
K
Faculty of Pharmaceutical Sciences, Tohoku Pharmaceutical University, 4-1 Komatsushima 4-chome, Aoba-ku, Sendai, Miyagi 981-8558, Japan.
Miyashiro
Yoshimichi
Y
Maekubo
Hitoe
H
Okuyama
Mitsunobu
M
Honma
Seijiro
S
Takahashi
Madoka
M
Numazawa
Mitsuteru
M
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001774:Blood Chemical Analysis / Q000379:methods*; D002138:Calibration; D002853:Chromatography, Liquid; D013196:Dihydrotestosterone / Q000032:analysis* / Q000097:blood* / Q000737:chemistry; D004952:Esters / Q000737:chemistry; D005260:Female; D006801:Humans; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D010848:Picolinic Acids / Q000737:chemistry; D011467:Prostate / Q000166:cytology*; D013048:Specimen Handling; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D013739:Testosterone / Q000032:analysis* / Q000097:blood* / Q000737:chemistry; D013997:Time Factors; D055815:Young Adult
D004952:Esters; D010848:Picolinic Acids; D013196:Dihydrotestosterone; D013739:Testosterone; C030614:picolinic acid