Total Steroid Concentrations Research Articles

Occurrence and Fate of Steroid Hormones in Sewage Treatment Plants

Sewage treatment plants (STPs) are one of the crucial barriers for the environmental emission of steroid hormones. Insights into the occurrence and fate of different categories of steroid hormones in STPs could provide theoretical support for improving steroid removal by STPs. The present study investigated 22 steroid hormones in each treatment process of two STPs located in Wuxi via eight monthly sampling campaigns and compared the efficacy of Anaerobic-Anoxic-Aerobic (A2/O) and reversed A2/O treatments. The results showed that the total steroid concentrations in the influent and effluent were 27.7-256.8 ng·L-1 and 5.7-211.0 ng·L-1, respectively, and 36.3-123.6 ng·g-1 in the excess sludge. Androsterone, androstenedione, estrone, estriol, and progesterone were the main species detected in the STPs. The concentrations of most steroids increased with the rise of rainfall and temperature, whereas the removal rates were not significantly different between winter and summer. Secondary and tertiary treatment processes showed better performance in steroid removal compared with that in the primary treatment; however, reversed A2/O did not show advantages over traditional A2/O. The organic-normalized partition coefficients (lg Koc) of steroids ranged between 2 and 4.5. The values of lg Koc in STP A were slightly greater than those in STP B, indicating that the partition behavior of steroids may influence their treatment efficacies.

Development of a Rapid, Targeted LC-IM-MS Method for Anabolic Steroids.

Anabolic steroids are of high biological interest due to their involvement in human development and disease progression. Additionally, they are banned in sport due to their performance-enhancing characteristics. Analytical challenges associated with their measurement stem from structural heterogeneity, poor ionization efficiency, and low natural abundance. Their importance in a variety of clinically relevant assays has prompted the consideration of integrating ion mobility spectrometry (IMS) into existing LC-MS assays, due primarily to its speed and structure-based separation capability. Herein we have optimized a rapid (2 min) targeted LC-IM-MS method for the detection and quantification of 40 anabolic steroids and their metabolites. First, a steroid-specific calibrant mixture was developed to cover the full range of retention time, mobility, and accurate mass. Importantly, this use of this calibrant mixture provided robust and reproducible measurements based on collision cross section (CCS) with interday reproducibility of <0.5%. Furthermore, the combined separation power of LC coupled to IM provided comprehensive differentiation of isomers/isobars within 6 different isobaric groups. Multiplexed IM acquisition also provided improved limits of detection, which were well below 1 ng/mL in almost all compounds measured. This method was also capable of steroid profiling, providing quantitative ratios (e.g., testosterone/epitestosterone, androsterone/etiocholanolone, etc.). Lastly, phase II steroid metabolites were probed in lieu of hydrolysis to demonstrate the ability to separate those analytes and provide information beyond total steroid concentration. This method has tremendous potential for rapid analysis of steroid profiles in human urine spanning a variety of applications from developmental disorders to doping in sport.

Concentrations and Composition of Sphagnum Peat Lipids Depending on Temperature in the Climatic Zones of Western Siberia

The composition of n-alkanes; sesqui-, di-, and triterpenoids; steroids; carboxylic acids; and tocopherols of sphagnum peat from the southern, middle, and northern taiga and forest-tundra of Western Siberia was studied by gas chromatography–mass spectrometry. It was shown that the latitudinal shift to the north leads to the preservation of the peat lipid components and sufficiently high values of carbon preference index (CPI) close to those of mire plants. This can be caused by a decrease in microbiological activity due to a decrease in the mean annual temperature. Towards the north, the total concentrations of n-alkanes, steroids, pentacyclic triterpenoids, squalene, and tocopherols increased, and a ratio of the total concentration of steroids to the concentrations of pentacyclic triterpenoids and hopanes decreased.

Three classes of steroids in typical freshwater aquaculture farms: Comparison to marine aquaculture farms

This study provides a comprehensive analysis of the occurrence of androgens, glucocorticoids and progestogens in typical freshwater aquaculture farms in comparison with marine aquaculture farms. The results showed that more steroids were detected in the marine aquaculture farms. For all aquatic products, the total concentrations of steroids in fish muscle were 22–2000ng/g, which were much higher than those detected in crabs, shrimps or mollusks (0.5–8.1ng/g). Based on the hazard index of the selected steroids, most water samples were in the low or medium risks. Some banned steroids were identified in the freshwater shrimps and all marine products, indicating that there may be certain health risks by the consumption of these aquatic products in the studied aquaculture farms. Furthermore, the total contributions of steroids in China were estimated to be 2300kg/y and 1200kg/y from freshwater fish culture and shrimp-crab culture, respectively.

Steroid bioaccumulation profiles in typical freshwater aquaculture environments of South China and their human health risks via fish consumption

More attention was previously paid to adverse effects of steroids on aquatic organisms and their ecological risks to the aquatic environment. So far, little information has been reported on the bioaccumulative characteristics of different classes of steroids in cultured fish tissues. The present study for the first time provided a comprehensive analysis of the occurrence, bioaccumulation, and global consumers’ health risks via fish consumption of androgens, glucocorticoids and progestanges in typical freshwater cultured farms in South China. The numbers and total concentrations of steroids detected in the tissues of five common species of the cultured fish were in the order of plasma > bile > liver > muscle and plasma > bile, muscle > liver, respectively. The field bioaccumulation factors for the detected synthetic steroids ranged from 450 to 97,000 in bile, 450 to 65,000 in plasma, 2900 to 16,000 in liver, and 42 to 2600 in muscle of fish, respectively. This data suggests that steroids are bioaccumulative in fish tissues. Mostly important, 4-androstene-3,17-dione (AED) and cortisone (CRN) were found to be reliable chemical indicators to predict the levels of steroids in plasma and muscle of the inter-species cultured fish, respectively. Furthermore, the maximum hazard quotients (HQs) of testosterone and progesterone were 5.8 × 10−4 and 9.9 × 10−5, suggesting that human health risks were negligible via ingestion of the steroids-contaminated fish.

Occurrence and sources of polar lipid tracers in sediments from the Shatt al-Arab River of Iraq and the northwestern Arabian Gulf

Shallow surface sediment samples from the southern part of the Shatt al-Arab River estuary of Iraq and the northwestern Arabian Gulf were analyzed for polar lipid compounds including n-alkanoic acids, n-alkanols, steroids and triterpenoids. The results showed that the n-alkanoic acids, methyl n-alkanoates and n-alkanols typically ranged from C12 to C32 with total concentrations of 3.2 to 108.2μgg−1dwt sample, from C12 to C30 with totals of 1.1 to 18.9μgg−1dwt sample, and from C14 to C32 at 1.8 to 112.6μgg−1dwt sample, respectively. Steroids and triterpenoids were detected and included stenols, stanols, stenones, stanones, tetrahymanol, tetrahymanone and extended ββ-hopanes. The total steroid concentrations ranged from 2.8 to 78.4μgg−1dwt sample, whereas the triterpenoids varied from 0.05 to 7.6μgg−1dwt sample. The simple regression analysis of the results and the spatial distribution patterns of the identified organic tracers indicated that the inter-compound relationships were related mainly to their major sources. Cluster analysis and principal component analysis (PCA) of data set showed that the sampling sites are similar. These sources were allochthonous (terrestrial vegetation), autochthonous (plankton residues and bacteria in the sediments) and anthropogenic (sewage and petroleum).

Compositions and sources of extractable organic matter in Mesopotamian marshland surface sediments of Iraq: II. Polar compounds

The concentrations of polar organic compounds including n-alkanoic acids, n-alkanols, steroids and triterpenoids were determined in extracts of shallow sediments from the Mesopotamian marshlands of Iraq. The sediments were collected by a stainless steel sediment corer, extracted with a dichloromethane and methanol mixture (3:1 v:v) by ultrasonic agitation and then analyzed by gas chromatography–mass spectrometric (GC–MS). The analysis results showed that the n-alkanoic acids ranged from C8 to C20 with concentrations of 7.8 ± 1.2 μg/g sample, whereas the concentrations of n-alkanols, which ranged from C12 to C39 were from 28.6 ± 4.3 to 121.7 ± 18.3 μg/g sample. The steroids and triterpenoids included stenols, stanols, stenones, stanones, tetrahymanol, tetrahymanone and extended ββ-hopanes. The total concentrations of steroids and triterpenoids ranged from 26.8 ± 4.1 to 174.6 ± 26.2 μg/g and from 0.74 ± 0.11 to 11.2 ± 1.7 μg/g sample, respectively. The major sources of these lipids were from natural vegetation, microbial (plankton) residues and bacteria in the sediments, with some contribution from anthropogenic sources (livestock, sewage and petroleum). Further studies of these wetlands are needed to characterize the input rate, transformation and diagenesis of the organic matter and to assess its various sources.

Fecal steroids in diarrhea II. Travellers' diarrhea

Fecal bile acid and neutral sterol patterns of five healthy adult volunteers from the United States who contracted travellers' diarrhea in Mexico City in 1974 were studied. The only pathogen in four of these patients was the heat-stable-only enterotoxin producing strain of Escherichia coli. Stool culture for pathogens was negative in the fifth subject. Bacterial 7α-dehydroxylation of cholic and chenodeoxycholic acids was not altered in diarrhea associated with toxigenic E. coli heat-stable, although total concentrations of bile acids in the feces in mg/g of feces (wet weight) were decreased during the diarrheal episode (1.3 ± 0.2 versus 3.3 ± 0.4 in controls, P < 0.02). Consistent with findings on bile acids, no significant changes in the production of coprostanol from cholesterol were observed. Although total concentrations of cholesterol metabolites, in milligrams per grams of feces (wet weight), were reduced in diarrhea, the difference was not significant statistically (2.2 ± 0.9 versus 5.4 ± 0.8 in controls, P > 0.05). There was an apparent increase in bacterial alteration of fecal steroids in the subject who showed no pathogens in stool culture, despite a decrease in total steroid concentrations of a magnitude similar to those observed in diarrhea associated with E. coli heat-stable. The production of coprostanol was correlated with 7α-dehydroxylation of cholic acid (r = 0.829, P < 0.001) and chenodeoxycholic acid (r = 0.749, P < 0.001). The results from this study are in striking contrast to the changes in gastrointestinal steroid metabolism previously observed in acute shigellosis.

Identification and quantitative determination of steroids in bovine corpus luteum during oestrous cycle and pregnancy.

Neutral steroids in bovine corpus luteum were isolated by liquid-gel chromatography on hydrophobic Sephadex, and were analyzed by computerized gas chromatography-mass spectrometry. The presence of progesterone and 20beta-hydroxy-4-pregnen-3-one was confirmed. In addition, 3beta-hydroxy-5-pregnen-20-one, 5-pregnene-3beta,20beta-diol, 3beta-hydroxy-5alpha-pregnan-20-one and 5alpha-pregnane-3beta,20beta-diol were fully identified, and 3-hydroxy-4-pregnen-20-one, 4 pregnene-3,20-diol, 22-hydroxycholesterol and 20,22-dihydroxycholesterol were partially characterized. Steroid sulphates were not detected. Quantification of the six fully identified steroids was based on peak areas in specific fragment ion current chromatograms constructed by the computer. During the 4th-19th day of the oestrous cycle the steroid concentrations varied as follows: progesterone 6.0-36.7 mug/g wet luteal weight, 20beta-hydroxy-4-pregnen-3-one 0.8-5.5 mug/g, 3beta-hydroxy-5-pregnen-20-one 1.0-7.1mug/g, 5-pregnene-3beta,20beta-diol smaller than 0.2-0.9 mug/g, 3beta-hydroxy-5alpha-pregnan-20-one 1.7-8.6 mug/g, and 5alpha-pregnane-3beta,20beta-diol smaller than 0.2-1.2 mug/g. The concentrations of progesterone and 3beta-hydroxy-5-pregnen-20-one seemed to vary in parallel and were low during days 11-17. During this period the concentrations of 5-pregnene-3beta,20beta-diol and 5alpha-pregnane-3beta,20beta-diol were highest as was the relative contribution of all three 20beta-hydroxysteroids to the total amount of steroids. The relative amount of 3beta-hydroxy-5alpha-pregnan-20-one seemed to be highest during days 4-6. The total steroid concentration in corpora lutea taken in early pregnancy (75-105 days) was 18-47 mug/g. In the period 75-90 days, progesterone constituted only 35-42% of the total steroids, 3beta-hydroxy-5alpha-pregnan-20-one as much as 23-40% and the total 20beta-hydroxysteroids 18-30%. The total steroid concentration in corpora lutea taken in midterm and late pregnancy was 21-77 mug/g. In this period progesterone was by far the predominant steroid and constituted about 80-90% of the total steroids in corpora lutea taken between days 150 and 240. Possible correlations between luteal growth, steroid oxidoreductases and steroid concentrations are discussed.

Endogenous neutral steroids in the lung tissue of early and mid-term human foetuses and in vitro studies on their formation.

ABSTRACT Four pools of lung tissue from 2 or 3 foetuses of 11–17 weeks' gestational age were analyzed by gas-liquid chromatography and gaschromatography-mass spectrometry for endogenous neutral steroids. All of the steroids detected were present as their mono- or disulphate conjugates. No free compounds were found. Dehydroepiandrosterone, pregnenolone and their 16α-hydroxylated derivatives were the compounds present in highest concentrations in the monosulphate fraction. The most prominent steroid in the disulphate fraction was 5-androstene-3β,17α-diol. Other compounds detected were the monosulphates of 3β,7α-dihydroxy-5-androsten-17-one, 3β,16β- dihydroxy -5- androsten-17-one, 3β,17β- dihydroxy-5-androsten-16-one, 5-androstene-3β,16β,1 7α-triol, 5-pregnene-3β,17α-diol, and 3β,17α-dihydroxy-5-pregnen-20-one and the disulphates of 5-androstene-3β,17β-diol, 3β,16α-dihydroxy-5-androsten-17-one and 5-pregnene-3β,20α-diol. The total concentrations of steroids in the lung tissue varied from 300 to 600 μg/100 g tissue wet weight. In order to gain some insight into the origin of the endogenous steroids detected, i. e. whether they were synthesized by the lung tissue or only filtered by it from the perfusing blood, minced lung tissue was incubated with dehydroepiandrosterone, pregnenolone, 3β,17α-dihydroxy-5-pregnen-20-one and with their sulphate conjugates. Evidence for the presence of the following enzymes was found: sulphokinase, 7α-hydroxylase, 16αhydroxylase, 17α-hydroxylase, 17β-hydroxysteroid dehydrogenase, 20α-hydroxysteroid dehydrogenase and C17–20 desmolase.

Serum protein-steroid hormone interactions I. Binding of deoxycorticosterone to serum proteins assayed in a metabolic system in vitro

Interaction between crystalline bovine serum albumin and deoxycorticosterone has been determined in a bioassay system in vitro involving the steroid-sensitive net influx of α-amino[1- 14C]isobutyrate into rat thymic lymphocytes. In this system, at a molar ratio of bovine serum albumin to deoxycorticosterone of 1:11, the inhibitory effect of the steroid was eliminated. Because of the unknown binding capacity and affinity of thymocytes for deoxycosticosterone, a molar binding capacity value of eleven for serum albumin and deoxycorticosterone must be considered a maximum value under the present experimental conditions. Cohn plasma protein fraction IV-4 had only a small capacity for binding deoxycorticosterone, whereas fractions IV-1, III and II were ineffective. Palmitic acid, cortisol and corticosterone did not appear to complete with deoxycorticosterone for binding sites on bovine serum albumin. Since that fraction of the total steroid concentration that became protein-bound was metabolically inert in the bioassay employed, the present results support the hypothesis that serum protein-bound steroids are biologically inactive.

Ultrafiltration studies of steroid-protein binding.

SUMMARY A method is described for determination of the extent of protein binding of steroids by ultrafiltration after addition of radioactively-labelled steroid tracers of high specific activity. Results obtained with specifically labelled 14C-steroids did not differ significantly from those with steroids randomly labelled with tritium. A number of steroids have been studied with this technique. The 'S' shaped curve obtained with some corticosteroids by plotting percentage ultrafilterable against steroid added to the plasma confirmed the presence of the corticosteroid-binding protein which has greater affinity for steroid than albumin. This method of plotting the ultrafiltration data afforded a method of assessing the amount of binding protein (or the number of sites). Cortisone, 17-hydroxy-11-deoxycorticosterone, cortisol, and Δ1 cortisol showed this type of curve. Binding of aldosterone, progesterone and 17-ketosteroids occurred to the same extent in plasma as in 5% albumin. The ultrafilterable fraction of these steroids in plasma was constant over a wide range of total steroid concentration. Binding of testosterone was greater with plasma than with albumin, and did not decrease upon addition of large amounts of testosterone.