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Results for “Steroid profiling”

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PubMed2002

Plasma steroid profiles following follicle-stimulating hormone or equine chorionic gonadotropin injection in cows chronically treated with gonadotropin-releasing hormone agonist.

Plasma steroid profiles following follicle-stimulating hormone (FSH) or equine chorionic gonadotropin (eCG) injection were studied in chronically gonadotropin releasing hormone agonist (GnRH-A)-treated cows. Follicular development and irINH secretion were stimulated by FSH or eCG injection. The plasma concentrations of estradiol-17 beta (E(2)) and testosterone (T) were markedly increased following eCG injection. However, significant increases of the plasma E(2) and T concentrations were not detected in FSH-treated cows. Ovulation of developed follicles were depended on the hCG injection in both groups. These results show: 1) Follicular response to an exogenous gonadotropin is still remained, 2) Ovulation of developed follicles is induced by hCG injection and 3) FSH and eCG cause disparate plasma steroid profiles, under the influence of repeated GnRH-A treatment.

Animals↗

PubMed2001

Changes in androgenic steroid profile due to urine contamination by microorganisms: a prospective study in the context of doping control.

Urine contamination by microorganisms may affect the interpretation of urinalysis in different areas of clinical diagnosis. This is particularly relevant in doping control. A prospective study was designed to assess the effects of urine contamination by selected pathogens on the endogenous androgenic steroid profile. Pooled urine from a healthy male volunteer with standard steroid profile compared with reference values for the Caucasian population was sterilized by filtration and stored in sterile glass tubes. Aliquots were inoculated with known amounts of 15 different organisms (bacteria, fungi, and moulds) and incubated at 37 degrees C for 2 weeks. Different markers of urine contamination, such as pH, deconjugation of steroids, and metabolic by-products, were determined. Alkalization of urinary pH was not a reliable indicator of urine contamination as several organisms grew in this medium and no alteration of this parameter was found. In uncontaminated urine, less than 10% of steroid glucuronide conjugates were spontaneously hydrolyzed. Higher rates of hydrolysis for sulfate conjugates were found. An unconjugated fraction higher than 10% of the total amount of testosterone was a reliable indicator of urine contamination. However, microbial production of testosterone or epitestosterone was not detected. In contrast, a few organisms were able to synthesize 5alpha-androstanedione, 5beta-androstanedione, and androstenedione using endogenous steroids as substrates.

Androgens↗

PubMed1986

[C21-steroid profiles in pregnancy associated with intrauterine growth retardation].

To study the steroidal profiles in pregnancy associated with IUGR, maternal serum steroids which included pregnenolone (P5), 16 alpha OH-P5, progesterone (P4) and 16 alpha OH-P4 levels were measured by RIA at 30-41 weeks of gestation and the levels were compared with those obtained from normal pregnancies. Thirty-three pregnancies were diagnosed as IUGR after delivery and 36 pregnancies were used as controls. The levels of P4 and 16 alpha OH-P4 were significantly lower in IUGR pregnancies than those in normal pregnancies, while the levels of P5 and 16 alpha OH-P5 tended to be higher in IUGR than in normal subject. A significant difference was observed between the 16 alpha OH-P4/16 alpha OH-P5 ratio in IUGR and normal pregnancy. When the levels of 16 alpha OH-P5 and 16 alpha OH-P4 were taken into account, 14 of 17 cases of IUGR were discriminated from normal subjects. These results demonstrated for the first time the clinical importance of measuring 16 alpha OH-C21 steroids in case of IUGR. Low delta4/delta5 steroid ratios also indicate that the placental 3 beta HSD activity might be impaired in IUGR pregnancy.

17-alpha-Hydroxypregnenolone↗

PubMed2002

A relatively simple and rapid multi-component method for analysis of steroid profiles in blood, fecal and liver samples.

Various methods of steroid analysis were assessed using radiolabeled steroids and thin layer chromatography. Three reversed phase chromatography systems were evaluated for separation and recovery of steroids extracted from blood, liver tissue and feces. The use of different numbers of Sep-Pak C(18) cartridges for the purification of steroid extracts was examined and steroid recoveries were measured and compared. The results indicated that recoveries were best when 4-6 cartridges were used. Rapid and slow procedures of enzymatic hydrolysis and acidic solvolysis of steroid conjugates were compared. A new and relatively rapid method for analysis of steroid profiles in liver, blood and fecal samples was developed. Assessment of this method showed that steroid recoveries were improved compared to existing methods with percentage recoveries of 64.1-82.5 for liver samples, 55.2-75 for blood samples, and 65.1-76.3 for fecal samples.

Cadaver↗

PubMed1995

Heterogeneity of urinary steroid profiles in children with adrenocortical tumors.

The excretory patterns of urinary steroids determined by capillary gas chromatography in 11 children (aged 0.8-16.5 years) with adrenocortical tumors were established. In 8 patients the predominant clinical feature was virilization, in 3 others, Cushing's syndrome. In 5 patients (3 carcinoma, 2 adenoma) very high excretion of 3 beta-hydroxy-5-ene steroids was observed. In 2 others (adenomas) only moderately elevated excretion of 11 beta-hydroxyandrosterone was found. In 1 patient (adenoma) pregnanediol dominated in the steroid profile, accompanied by moderately elevated 3 beta-hydroxy-5-ene steroids. Out of 3 Cushingoid patients (1 carcinoma, 2 adenomas), 1 presented an atypical urinary steroid pattern for hypercortisolemia, without 5 alpha-reductase and 11 beta-hydroxysteroid dehydrogenase deficiencies. Neither the urinary steroid pattern nor tumor size alone were reliable indicators of tumor malignancy, as evaluated by a pathological examination and subsequent metastasis-free survival.

Adenoma↗

PubMed2003

Improved specificity of newborn screening for congenital adrenal hyperplasia by second-tier steroid profiling using tandem mass spectrometry.

BACKGROUND: Newborn screening for congenital adrenal hyperplasia (CAH) involves measurement of 17alpha-hydroxyprogesterone (17-OHP), usually by immunoassay. Because this testing has been characterized by high false-positive rates, we developed a steroid profiling method that uses liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure 17-OHP, androstenedione, and cortisol simultaneously in blood spots. METHODS: Whole blood was eluted from a 4.8-mm (3/16-inch) dried-blood spot by an aqueous solution containing the deuterium-labeled internal standard d(8)-17-OHP. 17-OHP, androstenedione, and cortisol were extracted into diethyl ether, which was subsequently evaporated and the residue dissolved in LC mobile phase. This extract was injected into a LC-MS/MS equipped with pneumatically assisted electrospray. The steroids were quantified in the selected-reaction monitoring mode by use of peak areas in reference to the stable-isotope-labeled internal standard. We analyzed 857 newborn blood spots, including 14 blood spots of confirmed CAH cases and 101 of false-positive cases by conventional screening. RESULTS: Intra- and interassay CVs for 17-OHP were 7.2-20% and 3.9-18%, respectively, at concentrations of 2, 30, and 50 microg/L. At a cutoff for 17-OHP of 12.5 microg/L and a cutoff of 3.75 for the sum of peak areas for 17-OHP and androstenedione divided by the peak area for cortisol, 86 of the 101 false-positive samples were within reference values by LC-MS/MS, whereas the 742 normal and 14 true-positive results obtained by conventional screening were correctly classified. CONCLUSION: Steroid profiling in blood spots can identify false-positive results obtained by conventional newborn screening for CAH.

Adrenal Hyperplasia, Congenital↗

PubMed1988

Steroid profiling in the study of rat testicular steroidogenesis.

Twenty authentic steroids, derivatized as O-methyl oximes (MO), trimethylsilyl (TMS) ethers or as MO-TMS ethers have been subjected to capillary gas chromatography using two different columns. Virtually all of the steroid derivatives have been resolved, one difficult pair to separate being 5,16-androstadien-3 beta-ol and 5 alpha-androst-16-en-3 beta-ol on the non-selective phase OV-1. Where syn and anti forms of MO derivatives arose, these were also resolved under the conditions utilised. This technique of 'steroid profiling' has been applied to the separation and quantification of metabolites of pregnenolone which were formed during incubations of the microsomal and cytosolic fractions from rat testes. The majority of the metabolites were found in the microsomal incubation. These compounds included some odorous 16-androstenes as well as other C21 and C19 steroids, the formation of which was consistent with the 5-ene and 4-ene pathways of testosterone biosynthesis being operative. In addition, evidence was obtained for 16 alpha-hydroxylation of C21 steroids. Very much less metabolic activity was found in the cytosolic fraction of rat testes. Metabolic pathways have been proposed which both confirm and extend earlier work. We conclude that the rat testis can only form some of the odorous, possibly pheromonal, 16-androstenes and that these are quantitatively less important than in the porcine testis.

Animals↗

PubMed1983

Effect of vasectomy on the steroid profile of human seminal plasma.

The effect of vasectomy on the steroid profile of seminal plasma was studied in 19 healthy men at 4-monthly intervals during a 2-year post-vasectomy period. The steroids analyzed by radioimmunoassays were: androstenedione (A), testosterone (T), dihydrotestosterone (DHT), oestradiol (E2) and the sulphoconjugated forms of pregnenolone (delta 5P-S), dehydroepiandrosterone (DHEA-S), testosterone (T-S), dihydrotestosterone (DHT-S), oestrone (E1-S) and oestradiol (E2-S). The quantitatively most important steroids present in the seminal plasma were DHEA-S, delta 5P-S and DHT-S. Vasectomy resulted in a significant but non-progressive reduction in DHT (40%) and T (23%) levels. The levels of E2, delta 5P-S and DHT-S also exhibited a smaller reduction following vasectomy. No significant changes were observed in the levels of the other steroids studied. The results are interpreted as suggesting that under normal conditions the bulk of the steroids present in the ejaculate is contributed by the accessory reproductive organs. In addition, a significant part of the DHT and T reach the ejaculate together with the sperm directly from the testis and/or epididymis. It is suggested that the normal steroid levels in seminal plasma established in this investigation could serve as reference values in the hormonal evaluation of male infertility.

Adult↗

PubMed1986

Profiling steroid hormones and urinary steroids.

This paper reviews techniques utilized in the profiling of steroids in body fluids and tissues. Methods for profiling plasma unconjugated steroids and urinary steroid metabolites are focused on. Concentrations or levels of excretion of a variety of steroids have been documented and reviewed. The importance of profiling techniques in the study of normal and pathophysiology of hormonal steroids is discussed.

Animals↗

PubMed1983

Steroid profiles of body fluids other than urine, obtained by capillary gas chromatography.

Steroid profiling by capillary gas chromatography has been applied to the analysis of free and conjugated steroids in serum, ovarian follicles, lutein cyst and peritoneal fluids. Sep-Pak C18 octadecylsilica was used for extraction, whereas the separation of free and different conjugates was accomplished by successive hydrolysis and liquid extraction steps.

Ascitic Fluid↗

PubMed1995

Chemometric evaluation of urinary steroid profiles in doping control.

Ten endogenous steroid hormones and metabolites were determined according to the screening procedure for anabolic steroids in spot urine samples from 105 healthy young male athletes (control samples) and 23 males that tested positive for anabolic steroids in the doping control (positive samples). The GC-MS peak areas for each sample were normalized to total area. Multivariate data analysis by Partial Least Square Regression (PLSR) and using a coded Y-variable (positive samples: +1 and control samples -1) allows projection of the most systematic profile structures into a 2D plot revealing a clear distinction between the control and misuser groups. The most important determinants of the location in the loading plot were the ratios of testosterone to epitestosterone and androsterone to etiocholanolone. The ratio between 11-beta-hydroxyandrosterone and 11-beta-hydroxy-etiocholanolone was less important, in accordance with the fact that anabolic-androgenic steroid intake primarily affects the excretion of testosterone from the testis and to a much lesser degree adrenal steroid genesis. We present a preliminary validation of this model (PLS1-DISCRIM) for analysing steroid profiles in doping control samples from several categories of athletes, some of which are suspected for drug misuse, and results from a one dose excretion study in healthy volunteers. Our findings suggest that use of multivariate PLS-regression may give valuable information about anabolic androgenic steroid misuse in sport. When appropriately calibrated, this methodology may delineate drug misusers directly from the screening procedure for anabolic steroids in spot urine tests.

Adolescent↗

PubMed1983

Analysis of urinary steroid profiles of women with Cushing's syndrome by computerised gas chromatography-mass spectrometry.

Urinary steroidal profiles were studied in five women with Cushing's syndrome and in two normal women that were chosen as controls, by means of gas chromatography and gas chromatography-mass spectrometry. Four patients presented ACTH-dependent adrenal hyperplasia and the last patient had an adrenocortical carcinoma. Steroids were analyzed in urinary extracts, as their respective trimethylsilyloximes and/or trimethylsilylderivatives. Qualitative and quantitative data about 36 urinary steroids were obtained. Three pituitary patients showed a well defined picture of "5-ene pathway" in adrenal function. The fourth patient depicted some primary deficiencies of corticosteroid biosynthesis that overshadowed most biochemical expressions of Cushing's disease. The patient with adrenal carcinoma developed a steroidal pattern resembling autonomous functioning of adrenal cortex inner zones, showing both "5-ene pathway" and "4-ene pathway" increase. This patient also had an unexpected excretion of a major metabolite of 18-hydroxycorticosterone, that did not correlate with parameters of aldosterone production. Tetrahydro-6-hydroxy-cortisol was determined in urine of two patients and original data about urinary cortoic acids in Cushing's syndrome are given. Peripheral reductive metabolism played the most important role in almost all patients. In turn, some oxidative metabolic pathways for cortisol were not specifically favoured in the cases of Cushing's disease here reported.

18-Hydroxycorticosterone↗

PubMed1991

A gas chromatographic method for the determination of neutral steroid profiles in urine, including studies on the effect of oxytetracycline administration on these profiles in men.

A capillary gas chromatographic method for the determination of 'total' metabolic profiles of urinary neutral steroids was developed. The method is based on anion exchange chromatographic separation and purification of monoglucuronide-, monosulphate- and double-conjugated neutral steroids on DEAE-Sephadex A-25 microcolumns and the final analysis of the individual steroids in these conjugate groups is carried out by capillary column gas-liquid chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The method was shown to provide a convenient and accurate determination of total metabolic profiles of neutral steroids in urine and thus, can be used for metabolic studies of steroids and for diagnostic purposes. In the present investigation the effect of a tetracycline antibiotic on the production and metabolism of neutral steroids in men was studied during a 5-day oral administration of oxytetracycline. The results showed that the influence of oxytetracycline on neutral steroids was minor and mainly restricted to the changes in urinary neutral steroid glucuronide excretion. Oxytetracycline decreased the mean daily excretion of total neutral steroid monoglucuronides by 20% and a statistically significant decrease was found in the urinary excretion of 3 alpha-hydroxy-5 beta-androstan-17-one-glucuronide (etiocholanolone, 31%, P less than 0.05), 5 beta-pregnan-3 alpha,20 alpha-diolglucuronide (pregnanediol, 32%, P less than 0.05) and corticosteroid glucuronides, including 3 alpha,11 beta,17 alpha,20 beta,21-pentahydroxy-5 beta-pregnan- and 3 alpha,17 alpha,20 beta,21-tetrahydroxy-5 beta-pregnan-11-one-glucuronides (beta-cortol and beta-cortolone, 36%, P less than 0.05). The reason for this effect is unknown, but may be partly due to inhibition of intestinal hydrolysis of biliary steroid conjugates, which previously was shown to result in an interruption of enterohepatic circulation of steroids and an increased excretion of steroid conjugates by the faecal route.

Adrenal Cortex Hormones↗

PubMed1982

A semi-automated computerised gas-liquid chromatographic system for urinary steroid profile analysis.

A simple gas-liquid chromatographic method for the measurement of some of the major steroids in urine is described. The system uses conventional packed columns, automatic injection and is linked to an integrator. Calculations are all carried out by computer. The system is recommended as a simple screening procedure, producing a urinary steroid 'profile', which can process large numbers of urine samples cheaply and quickly, enabling patients worthy of more intensive investigation to be identified.

17-Hydroxycorticosteroids↗

PubMed1983

[A simple method for the estimation of a urinary steroids profile using glass capillary gas chromatography with a solventless injection system].

A simple method for the estimation of a urinary steroids profile with glass capillary gas chromatography is described. A 10ml portion of a urinary specimen was hydrolysed by Helix pomatia juice. Additional hydrolysis with ox-liver beta glucuronidase was necessary for the complete liberation of the urinary steroids from their glucuronides conjugates. Liberated free steroids were extracted with ethyl acetate, and the extract was washed and evaporated under reduced pressure. Finally extracted steroids were derivatised as MO-TMS (methoxime-trimethylsilyl) ethers. For the full-silylation of the steroids, the Sakauchi-Hornig method was employed and good results were obtained. An OV-101-WCOT column (30m length) was connected to the "solventless injection system" (a type of glass falling needle injection) and temperature-programmed gas chromatography was performed. By this method, eight fractions of 17-KS, Pregnanediol, Pregnanetriol, Pregnanetriolone, 5-Pregnentriol, tetrahydro-metabolites of 11-Deoxy-Cortisol and of Cortisol, and hexahydrometabolites of Cortisol were separated and quantitatively determined. Data processing was performed by an on-line microcomputer. The value in normal male and female controls was compatible with the results of glass capillary gas chromatography measured by Shackleton. Urine specimens from children with adrenocortical cancer and 21-hydroxylase deficiency were analyzed. Characteristic metabolic profiles of each patient were easily demonstrated. This method seems suitable for the routine clinical elucidation of abnormal steroid metabolism.

Adolescent↗

PubMed2001

Urinary steroid profile analysis.

BACKGROUND: A detailed analysis (profile) of the steroid metabolites in urine is useful for diagnosis of adrenal problems. Hospitals from many of UK health regions and around the world use the specialist assay and advisory service at UCLH. According to the total workload, samples are from patients with precocious puberty/premature adrenarche (21%), ambiguous genitalia (17%), Cushing's syndrome (13%), tumors (11%), polycystic ovaries (9%), hypertension (6%), problems of growth and development (5%), salt-loss (3%) and male pseudohermaphroditism (3%). Sixty percent of samples are from children and comprehensive reference data for steroid excretion rates in childhood unique to this laboratory were essential for interpretation of the results. CONCLUSION: The recognition and high excretion rates of certain steroids not easily measured in blood or urine by any other assays was particularly in cases of hypertension and tumors. The assay is cost effective by comparison with the combined costs of several individual hormone measurements but that cost may delay early referral to a specialist centre and that is not in the best interests of the families involved.

Adrenal Gland Diseases↗

PubMed1985

Measurement of urinary steroid profile in patients with adrenal tumor as a screening method for carcinoma.

Results of measurement of urinary steroid metabolite profile using gas chromatographic analysis in eight patients with adrenocortical tumors, i.e. 3 adenomas with Cushing's Syndrome, one adenoma with virilization, one adenoma without clinical manifestations, one carcinoma with Cushing's syndrome and virilization, one carcinoma with Cushing's syndrome and feminization, and one carcinoma without endocrinological symptoms, are reported. A unique pattern dominated by 5 beta and 11 beta-hydroxy steroid metabolites was confirmed in five patients with Cushing's syndrome consisting of three cases with adenomas and two with carcinomas. Excessive 3 alpha, 17 alpha, 21-trihydroxy-5 beta-pregnan-20-one (tetrahydro-11-deoxycortisol, THS) and delta 5-pregnene-3 beta, 11 alpha, 20 alpha-triol (delta 5-pregnenetriol) values were found in all three carcinomas including a nonfunctional carcinoma. These findings would strongly suggest the tumor to be a carcinoma, although excessive excretion of THS and delta 5-pregnenetriol was detected in one patient with a large adenoma associated with virilization. One patient with carcinoma was responsive to ACTH stimulation while the remainder show almost no response to exogenous ACTH. Urinary steroid profiling using gas chromatographic analysis, especially the values for THS and delta 5-pregnenetriol, appears to be a useful method to use in detecting these steroid metabolic characteristics in patients with adrenocortical carcinoma.

Adenoma↗

PubMed1978

Quantitative analysis of steroid profiles from urine by capillary gas chromatography. I. Accuracy and reproducibility of the sample preparation.

A method is described for the determination of steroid profiles from urine by means of gas chromatography using high-efficiency glass capillary columns. The accuracy and reproducibility of essential steps in the sample preparation (extraction of steroids and steroid conjugates by means of XAD-2, enzymatic hydrolysis with Helix pomatia juice, solvolysis in acidified ethyl acetate and alkali wash) are established using different endogenously labelled urine samples, obtained from normal subjects to whom labelled steroids had been administered. Preliminary results are given on the reproducibility of the derivatization procedure (formation of methoxime-trimethylsilyl (MO-TMS) ethers), the gas chromatographic analysis and the whole method. Two procedures for the purification of MO-TMS steroid derivatives are compared. Application of the method to urine samples of patients with various endocrine disorders is included.

Adult↗