Accurate determination of tissue steroid hormones, precursors and conjugates in adult male rat.
The actual levels of steroid hormones in organs are vital for endocrine, reproductive and neuronal health and disorders. We developed an accurate method to determine the levels of steroid hormones and steroid conjugates in various organs by an efficient preparation using a solid-phase-extraction cartridge. Each steroid was identified by the precursor ion spectra using liquid chromatography-electrospray ionization time-of-flight mass spectrometry, and the respective steroids were quantitatively analysed in the selected reaction monitoring mode by liquid chromatograph-mass spectrometry/mass spectrometry (LC-MS/MS). The data showed that significant levels of testosterone, corticosterone and precursors of both hormones were detected in all organs except liver. The glucuronide conjugates of steroid hormones and the precursors were detected in all organs except liver, but sulfate conjugates of these steroids were observed only in the target organs of the hormones and kidney. Interestingly, these steroids and the conjugates were not observed in the liver except pregnenolone. In conclusion, an accurate determination of tissue steroids was developed using LC-MS analysis. Biosynthesis of steroid hormones from the precursors was estimated even in the target organs, and the delivery of these steroid conjugates was also suggested via the circulation without any significant hepatic participation.
Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Hokkaido 069-8501, Japan.
Naoyuki
N
Maeda
Emi
E
Tanaka
Tomokazu
T
Suzuki
Kanako
K
Okumura
Sachiko
S
Nomura
Taku
T
Miyasho
Satoko
S
Haeno
Hiroshi
H
Yokota
D000311:Adrenal Glands / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003345:Corticosterone / Q000032:analysis* / Q000096:biosynthesis / Q000097:blood / Q000378:metabolism; D045166:Estradiol Congeners / Q000032:analysis* / Q000096:biosynthesis / Q000097:blood / Q000378:metabolism; D020719:Glucuronides / Q000737:chemistry / Q000378:metabolism; D007668:Kidney / Q000737:chemistry / Q000378:metabolism; D057230:Limit of Detection; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008297:Male; D009928:Organ Specificity; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization; D013431:Sulfates / Q000737:chemistry / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013737:Testis / Q000737:chemistry / Q000378:metabolism; D045165:Testosterone Congeners / Q000032:analysis* / Q000096:biosynthesis / Q000097:blood / Q000378:metabolism
D045166:Estradiol Congeners; D020719:Glucuronides; D013431:Sulfates; D045165:Testosterone Congeners; D003345:Corticosterone