AIMTo establish a method to determine the isotope ratios of 13C to 12C of dehydroepiandrosterone and its metabolites in urine, for detecting the source of dehydroepiandrosterone or its metabolites.

METHODSPreliminary separation of endogenous anabolic androgenic steroids could be achieved using solid phase extraction, enzymolysis and thin layer chromatography. The source of dehydroepiandrosterone and other endogenous anabolic androgenic steroids could be detected by their delta values with gas chromat ography-combustion-isotope ratio mass spectrometry.

RESULTSThe 5 values of some metabolites of dehydroepiandrosterone reduced after the administration of dehydroepiandrosterone preparation. In these cases the data indicated that exogenous anabolic androgenic steroids were administrated.

CONCLUSIONThe source of dehydroepiandrosterone or its metabolites in urine could be detected by measuring their delta values with this method.

Adult ; Androstane-3,17-diol ; urine ; Androsterone ; urine ; Chromatography, Thin Layer ; methods ; Dehydroepiandrosterone ; metabolism ; Doping in Sports ; Etiocholanolone ; urine ; Female ; Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Male ; Pregnanetriol ; urine ; Substance Abuse Detection ; methods

Congenital adrenal hyperplasia refers to a group of autosomal recessive disorders that is defective in the synthesis of cortisol. The enzymes most often affected are 21-hydroxylase and 11beta hydroxylase. The low levels of cortisol stimulate the pituitary gland to release ACTH. Chronic elevation of the ACTH level causes bilateral adrenal hyperplasia and a secondary increase in androgen formation. We examined a 19 year-old woman presented with clitoral hypertrophy and vaginal spotting. The subjects basal level of serum cortisol was low, but the serum levels of ACTH, 17a-hydroxyprogesterone, deoxy-corticosterone were elevated. The urinary excretions of 17-ketosteroids and 17-hydroxycorticosteroids were also increased. The karyotyping study and transrectal ultrasonography showed normal findings. The patient underwent clitoris reduction surgery and received hydrocortisone. To the best of our knowledge, this is the first case of 11beta-Hydroxylase deficiency in Korea.
17-Hydroxycorticosteroids ; 17-Ketosteroids ; Adrenal Hyperplasia, Congenital* ; Adrenocorticotropic Hormone ; Clitoris ; Female ; Humans ; Hydrocortisone ; Hyperplasia ; Hypertrophy ; Karyotyping ; Korea ; Metrorrhagia ; Pituitary Gland ; Steroid 21-Hydroxylase ; Ultrasonography ; Young Adult

OBJECTIVE: The aim of this study was to measure maternal plasma androgens, estrogen and leptin levels and to assess the role of these hormones in the pathogenesis of preeclampsia. METHODS: The groups consisted of 32 healthy pregnant women as well as 28 pregnant women with severe preeclampsia. Plasma leptin, total testosterone (T), estradiol (E2), dehydroepiandrosterone sulfate (DHEAS) and androstenedione (ADD) levels were measured. Statistical analysis was achieved with Student's t-test by using SPSS for Windows and the Pearson's coefficient of correlation was calculated. RESULTS: No significant differences were observed between the two groups regarding age, gestational age, body mass index, parity, hematocrit and platelet, whereas significant differences were noted regarding systolic and diastolic blood pressure, gestational weeks at delivery, birth weight, serum creatinine, uric acid and urea (p<0.05). In preeclampsia group, serum total testosterone and ADD levels were determined to be higher than the control group (p<0.05). However, there was no significant differences in plasma levels of DHEAS and E2 among the two groups. The plasma levels of leptin were not significantly increased in the preeclampsia group. Serum testosterone levels were positively correlated with systolic and diastolic pressure and uric acid and negatively correlated with birth weight. CONCLUSION: These results suggest that the elevated plasma levels of testosterone could contribute to the pathogenesis of preeclampsia.
Androgens ; Androstenedione ; Birth Weight ; Blood Platelets ; Blood Pressure ; Body Mass Index ; Creatinine ; Dehydroepiandrosterone ; Dehydroepiandrosterone Sulfate ; Estradiol ; Estrogens ; Female ; Gestational Age ; Gonadal Steroid Hormones* ; Hematocrit ; Humans ; Leptin* ; Parity ; Plasma* ; Pre-Eclampsia* ; Pregnant Women ; Testosterone ; Urea ; Uric Acid

OBJECTIVE: The aim of this study was to measure maternal plasma androgens, estrogen and leptin levels and to assess the role of these hormones in the pathogenesis of preeclampsia. METHODS: The groups consisted of 32 healthy pregnant women as well as 28 pregnant women with severe preeclampsia. Plasma leptin, total testosterone (T), estradiol (E2), dehydroepiandrosterone sulfate (DHEAS) and androstenedione (ADD) levels were measured. Statistical analysis was achieved with Student's t-test by using SPSS for Windows and the Pearson's coefficient of correlation was calculated. RESULTS: No significant differences were observed between the two groups regarding age, gestational age, body mass index, parity, hematocrit and platelet, whereas significant differences were noted regarding systolic and diastolic blood pressure, gestational weeks at delivery, birth weight, serum creatinine, uric acid and urea (p<0.05). In preeclampsia group, serum total testosterone and ADD levels were determined to be higher than the control group (p<0.05). However, there was no significant differences in plasma levels of DHEAS and E2 among the two groups. The plasma levels of leptin were not significantly increased in the preeclampsia group. Serum testosterone levels were positively correlated with systolic and diastolic pressure and uric acid and negatively correlated with birth weight. CONCLUSION: These results suggest that the elevated plasma levels of testosterone could contribute to the pathogenesis of preeclampsia.
Androgens ; Androstenedione ; Birth Weight ; Blood Platelets ; Blood Pressure ; Body Mass Index ; Creatinine ; Dehydroepiandrosterone ; Dehydroepiandrosterone Sulfate ; Estradiol ; Estrogens ; Female ; Gestational Age ; Gonadal Steroid Hormones* ; Hematocrit ; Humans ; Leptin* ; Parity ; Plasma* ; Pre-Eclampsia* ; Pregnant Women ; Testosterone ; Urea ; Uric Acid

OBJECTIVE: The aim of this study was to measure maternal plasma androgens and estrogen levels and to assess the role of these hormones in the pathogenesis of preeclampsia. METHODS: The groups consisted of 28 healthy pregnant women as well as 24 pregnant women with severe preeclampsia. Plasma total testosterone (T), estradiol (E2), dehydroepiandrosterone sulfate (DHEAS) and androstenedione (ADD) levels were measured. Statistical analysis was achieved with Student's t-test by using SPSS for Windows and the Pearson's coefficient of correlation was calculated. RESULTS: No significant differences were observed between the two groups regarding age, gestational age, body mass index, parity, hematocrit and platelet, whereas significant differences were noted regarding systolic and diastolic blood pressure, gestational weeks at delivery, birth weight, serum creatinine, uric acid and urea. In preeclampsia group, serum total testosterone and ADD levels were determined to be higher than the control group (p<0.05). However, there was no significant differences in plasma levels of DHEAS and E2 among the two groups (p>0.05). Serum testosterone levels were positively correlated with systolic and diastolic pressure and uric acid and negatively correlated with birth weight. CONCLUSION: These results suggest that the elevated plasma levels of testosterone could contribute to the pathogenesis of preeclampsia.
Androgens ; Androstenedione ; Birth Weight ; Blood Platelets ; Blood Pressure ; Body Mass Index ; Creatinine ; Dehydroepiandrosterone ; Dehydroepiandrosterone Sulfate ; Estradiol ; Estrogens* ; Female ; Gestational Age ; Hematocrit ; Humans ; Parity ; Plasma* ; Pre-Eclampsia ; Pregnancy ; Pregnancy Trimester, Third* ; Pregnancy* ; Pregnant Women ; Testosterone ; Urea ; Uric Acid

BACKGROUND: Hyperprolactinemia has been linked with hyperandrogenism and hirsutism in some women. High plasma Dihydroandrosterone and DHA-S levels were reported in patients with hyperprolactinemia and a dissociation of adrenal androgen and cortisol secretion occurs in normal subjects. The mechanism has not been elucidated, but it has been suggested that pituitary factors other than ACTH modulate adrenal androgen synthesis, One candidate hormone is prolactin. Adrenal tissue has been found to possess prolactin receptors and prolactin has been shown to act synergistically with ACTH and lowers the activity of the enzyme 5a-reductase or 3B-hydroxysteroid dehydrogenase (3B-HSD). The aim of this study was to investigate the secretion of adrenal androgen metabolites in patients with idiopathic hyperprolactinemia and prolactinoma and to deterrnine the relationship with prolactin and androgens. METHODS: We measured 24 hour-urinary DHEA, androstenedione, androsterone, pregnenolone, tetrahydrocorticoid and cortisol in 16 normal controls and 5 patients with idiopathic hyperprolac-tinemia (HP) and 12 patients with prolactonoma in the early follicular phase. RESULTS: Urinary DHEA, AD (androsteredione), and androsterone, the metabolites of adrenal androgen, were significantly higher in both patients with idiopathic HP and prolactinoma compared with those in normal controls (p<0.05), whereas they were not different in both disease groups. Urinary pregnenolone levels, early metabolite of adrenal steroid synthesis, were lower in patients. In contrast, urinary tetrahydorcortisol and cortisol were higher in patients compared to controls. There was no difference in DHEA:androsterone ratio between patients and controls. And there were no correlation between prolactin levels and the levels of androgenic metabolites or clinical symptoms. CONCLUSION: Prolactin has a tropic effct on the secretion of androgens and steroids by the adrenal cortex. But prolactin levels were not correlated with androgen levels or clinical symptoms (amenorrhea), and it might have little effect on lowering the activity of 3B-HSD.
Adrenal Cortex ; Adrenocorticotropic Hormone ; Androgens ; Androstenedione ; Androsterone ; Dehydroepiandrosterone ; Female ; Follicular Phase ; Hirsutism ; Humans ; Hydrocortisone ; Hyperandrogenism ; Hyperprolactinemia* ; Oxidoreductases ; Plasma ; Pregnenolone ; Prolactin ; Prolactinoma ; Receptors, Prolactin ; Steroids

Bilateral adrenal tuberculosis is a rare disease and often occurs bilaterally. We report a case of bilateral adrenal masses due to tuberculosis with adrenal insufficiency. The patient was a 39- year-old man who had complained of intermittent pain of right upper quadrant and general weakness. The plasma levels of cortisol and catecholamine were normal. The levels of 24-hour urinary catecholamine and VMA were also normal. But the levels of 24-hour urinary 17- hydroxycorticosteroids and 17-ketosteroids were reduced. Abdominal CT showed about 6.5 x4.8 x 5.4cm sized left adrenal mass and 4.0 x 2.8 x 3.6cm sized right adrenal mass with calcification. The result of sono-guided percutaneous needle biopsy was adrenal tuberculosis. The patient was treated with antituberculous chemotherapy and hormonal replacement. But the masses are unchanged during 5-months follow-up.
17-Ketosteroids ; Adrenal Glands ; Adrenal Insufficiency ; Biopsy, Needle ; Drug Therapy ; Follow-Up Studies ; Humans ; Hydrocortisone ; Hydroxycorticosteroids ; Plasma ; Rare Diseases ; Tomography, X-Ray Computed ; Tuberculosis*

Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, β-estradiol, α-estradiol, testosterone, dehydroepiandrosterone, 17á-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities (R² > 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD < 2.43%), and recovery rates (97.3% to 104.6%) for all eleven SHs were determined. In addition, a method for the quantification of three 7-oxycholesterols (7-O-CSs: 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol) in the NZA was established by using an HPLC-photodiode array (PDA) method. The linearities (R² > 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.
Androsterone ; Animals ; Antlers ; Chromatography, Liquid ; Deer ; Dehydroepiandrosterone ; Estrone ; Medroxyprogesterone ; Megestrol Acetate ; Methods ; New Zealand ; Progesterone ; Steroids ; Testosterone

OBJECTIVETo assess the utility of serum steroids measurement in monitoring the treatment of children with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21OHD).

METHODNineteen Patients with CAH 21OHD aged (3.67±1.54) years treated with hydrocortisone and fluorocortisone replacement were followed up at an intervals of 0.33 - 1.0 years over a period of (1.47±0.7) years. At each visit, roentgenograms of the hands and wrists were taken, fasting peripheral blood were collected to test serum dehydroepiandrosterone sulfate, progesterone, 17-hydroxyprogesterone (17-OHP), androstenedione (Δ4-A), testosterone, free testosterone, estrone, and estradiol concentrations at 8 AM in the morning before the first dose of glucocorticoid. Then the patients were classified as being in "Good Control" or in "Poor Control" based on clinical criteria including signs of androgen excess, growth velocity and bone age increment at each interval. Comparisons were carried out between the serum steroid concentrations of the two groups. The receiver operating characteristic (ROC) curves were used to determine the cut-off values for diagnosing "Poor Control".

RESULTBoth of serum Δ4-A and 17-OHP concentrations were higher in "Poor Control" group than those in "Good Control" group [5.95 (2.23-11.2) nmol/L versus 1.05 (1.05-9.89) nmol/L, t=2.19; 13.85 (6.06-20) µg/L versus 3.67 (0.42-21.1) µg/L, t=2.17; P<0.05, respectively]. The ROC curves for serum Δ4-A concentrations, serum 17-OHP concentrations, serum Δ4-A in combination with 17-OHP concentrations were constructed with areas under the ROC curves (95%CI) of 0.76 (0.62, 0.90), 0.75 (0.62, 0.88), 0.69 (0.54, 0.84), P<0.05, respectively. Serum Δ4-A of 3.9 nmol/L had 0.78 of sensitivity and 0.75 of specificity in diagnosing "Poor Control". Serum 17-OHP of 7.1 µg/L has 0.67 of sensitivity and 0.71 of specificity in diagnosing "Poor Control".

CONCLUSIONEach of serum 17-OHP or/and Δ4-A concentration was of significance in diagnosing "Poor Control" during the glucocorticoid replacement treatment of CAH 21OHD, with the diagnostic efficacy being serum Δ4-A concentration, serum 17-OHP concentration and serum Δ4-A in combination with 17-OHP concentration in descending order. Serum Δ4-A and 17-OHP concentrations may be used as the biochemical indicators to monitor the therapy of CAH 21OHD.

17-alpha-Hydroxyprogesterone ; blood ; Adrenal Hyperplasia, Congenital ; blood ; diagnosis ; therapy ; Androstenedione ; blood ; Child, Preschool ; Dehydroepiandrosterone Sulfate ; blood ; Female ; Humans ; Hydrocortisone ; blood ; Male ; Progesterone ; blood ; Steroid 21-Hydroxylase ; blood ; Testosterone ; blood

The fragmentation pathways of five estrogens (estradiol, estrone, equilin sulfate, 17 a-dihydroequilin sulfate and equilenin sulfate) have been studied with high resolution and high mass accuracy using electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF/MS) in the negative ion mode. Molecular weights were obtained from [M-H](-) ions in the product ion spectra. The results indicate that the five structurally similar estrogens have similar fragmentation pathways. Using their stable isotope forms as internal reference compounds, the accurate mass and composition of the fragment ions were determined. During collision-induced dissociation (CID), cleavage is initiated by loss of oxygen atoms from carbon-17, after which D and C rings cleave sequentially and rearrange to finally form stable conjugate structures with highly abundant characteristic fragment ions at m/z 183 (accompanied by m/z 181), m/z 169 and m/z 145 (accompanied by m/z 143). Understanding these characteristic fragmentation pathways of estrogens will be helpful in identifying the structures of steroid hormones in general.
Chemical Fractionation ; methods ; Equilenin ; chemistry ; Equilin ; analogs & derivatives ; chemistry ; Estradiol ; chemistry ; Estrogens ; chemistry ; Estrone ; chemistry ; Ions ; Spectrometry, Mass, Electrospray Ionization