Analysis Of Steroids Research Articles

Long-term Hyperandrogenemia and/or Western-style Diet in Rhesus Macaque Females Impairs Preimplantation Embryogenesis.

Hyperandrogenemia and obesity are common in women with polycystic ovary syndrome, but it is currently unclear how each alone or in combination contribute to reproductive dysfunction and female infertility. To distinguish the individual and combined effects of hyperandrogenemia and an obesogenic diet on ovarian function, prepubertal female rhesus macaques received a standard control (C) diet, testosterone (T) implants, an obesogenic Western-style diet (WSD), or both (T + WSD). After 5 to 6 years of treatment, the females underwent metabolic assessments and controlled ovarian stimulations. Follicular fluid (FF) was collected for steroid and cytokine analysis and the oocytes fertilized in vitro. Although the T + WSD females exhibited higher insulin resistance compared to the controls, there were no significant differences in metabolic parameters between treatments. Significantly higher concentrations of CXCL-10 were detected in the FF from the T group, but no significant differences in intrafollicular steroid levels were observed. Immunostaining of cleavage-stage embryos revealed multiple nuclear abnormalities in the T, WSD, and T + WSD groups. Single-cell DNA sequencing showed that while C embryos contained primarily euploid blastomeres, most cells in the other treatment groups were aneuploid. Despite yielding a higher number of mature oocytes, T + WSD treatment resulted in significantly reduced blastocyst formation rates compared to the T group. RNA sequencing analysis of individual blastocysts showed differential expression of genes involved in critical implantation processes between the C group and other treatments. Collectively, we show that long-term WSD consumption reduces the capacity of fertilized oocytes to develop into blastocysts and that the addition of T further impacts gene expression and embryogenesis.

A tandem liquid chromatography and tandem mass spectrometry (LC/LC–MS/MS) technique to separate and quantify steroid isomers 11β-methyl-19-nortestosterone and testosterone

Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has become a mainstay analytical technique in pharmaceutical research and development and clinical diagnosis due to several advantages including excellent selectivity, specificity, and high sensitivity. LC-MS/MS has become the method of choice for steroids analysis due to its fast analytical time and improved specificity yet has a challenge in the separation and measurement of isomers with the same product ions. Here we describe a high-sensitivity LC/LC-MS/MS method that combines chiral chromatography and reverse-phase chromatography (LC/LC) along with MS/MS to rapidly separate and quantify steroid isomers of 11ß-methyl-19-nortestosterone (11ß-MNT) and endogenous testosterone in serum.

Parallel targeted and non-targeted quantitative analysis of steroids in human serum and peritoneal fluid by liquid chromatography high-resolution mass spectrometry

We developed and validated a liquid chromatography high-resolution mass spectrometry method for the absolute quantification of 51 steroids for clinical analysis of human serum and, for the first time, peritoneal fluid. Data acquisition was performed in both targeted and untargeted mode simultaneously, thus allowing the accurate and precise quantification of the main components of the classical steroid pathways (17 steroids) as well as the analysis of 34 additional non-classical steroids. For targeted analysis, validation was performed according to FDA guidelines, resulting, among other parameters, in accuracy < 13% RSD and precision < 10% relative error, for both inter- and intra-day validation runs. By establishing steroid-specific response factors, the calibration curves of the targeted analytes can be extended to untargeted analytes. This approach opens novel possibilities for the post hoc analysis of clinical samples as the data can be examined for virtually any steroid even after data acquisition, enabling facile absolute quantification once a standard becomes available. We demonstrate the applicability of the approach to evaluate the differences in steroid content between peripheral serum and peritoneal fluid across the menstrual cycle phases, as well as the effect of the synthetic gestagen dienogest on the steroid metabolome.Graphical abstract

Small Blubber Samples (50 mg) Sufficient for Analyses of 10 Stress and Reproductive Steroid Hormones in Gray and Fin Whales via Liquid Chromatography Mass Spectrometry

Information on stress, reproductive fitness, and health is difficult to obtain in wild cetaceans but critical for conservation and management. The goal of this study was to develop a methodology requiring minimal blubber mass for analysis of reproductive and stress steroid hormones and, hence, suitable for cetacean biopsies. Blubber biopsies and samples were collected from free-ranging and stranded gray and fin whales. Steroid hormones were extracted from blubber samples as small as 50 mg using liquid-liquid extraction methodology developed to handle the high fat content of blubber. Samples were analyzed via liquid chromatography with tandem mass spectrometry for 10 hormones: aldosterone, androstenedione, cortisol, cortisone, corticosterone, 17β-estradiol, estrone, 17α-hydroxyprogesterone, progesterone, and testosterone. As part of the optimization, homogenization via bead beating and blade dispersion were compared, and the former found superior. To investigate optimal yet minimal tissue mass required, hormone panels were compared among paired 50, 150, and 400 mg samples, the latter two being commonly reported masses for hormone blubber analysis. Results indicated that 50 mg of blubber was suitable and sometimes superior. Additionally, significant differences in precision values were observed between species, possibly stemming from differences in blubber composition, and relevant to homogenization technique selection and calibration methods that use blubber matrix matches obtained from a species other than the study species. Based on recovery and precision values, our methodology was accurate and precise in the measurement of spiked known quantities for all 10 hormones, confirming the methodology capabilities in 50 mg blubber mass in both species. Altogether, and in our specific sample sets, all endogenous hormones, except corticosterone, were identified above the detection limit in 50 mg gray whale blubber samples while all endogenous hormones, except aldosterone, cortisone, estrone, and progesterone, were detected in 50 mg fin whale blubber samples. We present a robust methodology for the analysis of multiple reproductive and stress steroid hormones in minimal masses of cetacean blubber compatible with small biopsies. Finally, we identified statistically significant differences in corticosteroid concentrations between stranded and free ranging animals.

Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) Method for the Quantification of Steroids Androstenedione, Dehydroepiandrosterone, 11-Deoxycortisol, 17-Hydroxyprogesterone, and Testosterone.

Congenital adrenal hyperplasia (CAH) is a group of autosomal-recessive disorders due to deficiency of 11- or 21-hydroxylase. The analysis of cortisol, androstenedione, 17-hydroxyprogesterone (OHPG), dehydroepiandrosterone (DHEA), 11-deoxycortisol, and testosterone is generally performed in the diagnosis and/or follow-up of CAH. Analysis of specific steroids is also performed in other disorders such as evaluation of hirsutism or infertility in females and hypogonadism in males. Cortisol is generally analyzed by immunoassays, whereas other hormones are preferably assayed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A multiple reaction monitoring, positive mode atmospheric pressure chemical ionization, LC-MS/MS method is described for the simultaneous quantification of androstenedione, 17-hydroxyprogesterone, DHEA, 11-deoxycortisol and testosterone. The method involves addition of labeled internal standards to serum samples and extraction of steroids in methyl tert-butyl ether. The extract is evaporated under stream of nitrogen, and the residue is reconstituted in methanol and analyzed by LC-MS/MS.

HPLC fractionation with immunoassay of steroids from nipple aspirate fluid

Fractionation of steroids allows for multiple assays to be run on a single low volume liquid biopsy, whereas performing the same number of assays without fractionation would require increasing the sample volume by dilution, rendering the concentration of steroids below the level of detection for most, if not all, downstream assays. Briefly, steroids are extracted from a biofluid sample using solvent phase extraction to separate the aqueous (conjugated) steroids from the non-aqueous (non-conjugated) steroids in the organic phase. The latter is further separated by high-performance liquid chromatography (HPLC) and collected in an automated fraction collector based on the UV detection of internal standards. Commercially available immunoassays are then used to quantify the < ng/ml concentrations of steroids in each fraction. This protocol was designed for small samples of nipple aspirate fluid (minimum 2 µL), but it can be modified to fractionate steroids from homogenized solid tissue samples or other liquid biopsies. Included in this protocol are precautions to help ensure reproducibility and minimize matrix effects and other errors of measurement, given that samples requiring fractionation are fundamentally precious and, like other quantitative procedures of small samples, can be prone to contamination by solvent residues and other factors.•The method permits quantitative analysis of multiple steroids from very small volumes of biofluid.•Fractionation by HPLC provides a highly purified sample for quantification.•The immunoassay end point provides specificity without expensive equipment.

Clinical Analysis of Steroids Induced Glaucoma: A Systematic Review

For more than 50 years, the elevation in intraocular pressure that can impede the administration of topical or systemic corticosteroids has been recognized. There has been increasing attention in this steroid-responsive phenomena since the identification of the myocilin gene (formerly known as the trabecular meshwork inducible glucocorticoid response or TIGR gene). In addition, the increasingly popular use of injectable intraocular steroids to treat clinically substantial subretinal fluid and macular oedema has increased the incidence. Animal studies, cell biology, molecular biology, and a better grasp of genetics have all contributed to a better understanding of the response's mechanics. The electronic MEDLINE, Embase, Cochrane, and PubMed databases were searched. Additionally, the bibliography of all relevant articles and textbooks were manually searched. To determine the link between plasma cortisol levels and steroid-induced glaucoma, plasma cortisol levels can be tested once every four hours. It can also be done on a bigger group of people. The steroid provocative test will aid in the identification of individuals who are high steroid responders in the general population. It can also be used in patients who require intravitreal triamcinolone injections. Genetic research could shed further light on the aetiology of steroid-induced glaucoma.

Toward Routine Analysis of Anabolic Androgenic Steroids in Urine Using Ion Mobility-Mass Spectrometry.

Anabolic androgenic steroids (AAS) make up one of the most prevalent classes of performance-enhancing drugs banned by the World Anti-Doping Agency (WADA) due to the competitive advantage they can afford athletes. Mass spectrometry-based methods coupled with chromatographic separations have become the gold standard for AAS analysis because of the superior sensitivity and selectivity provided. However, emerging analytical techniques including ion mobility spectrometry (IMS) have been demonstrated in recent applications as a means to further characterize and identify potential unknowns while simultaneously delivering improved sensitivity by filtering noise. Herein we outline the next crucial steps in bringing IMS to the routine drug testing workflow by combining it with established chromatographic and mass spectrometry methods (i.e., LC-IM-MS) for the detection of AAS in human urine. In addition to robust measurement of collision cross sections which can be used for identification purposes, functional group microtrends provide a structural basis on which to elucidate the structure of future novel anabolic agents. Lastly, the developed workflow is tested by analysis of testosterone in a realistic matrix (human urine) and demonstrates a limit of detection of 524 pg/mL, which surpasses the WADA Minimum Required Performance Levels for anabolic steroids. This work is expected to pave the way toward routine incorporation of IMS into analytical drug testing workflows to augment both qualitative and quantitative measure of performance enhancing drugs in the future.

Effect of kelor (Moringa oleifera Lam) ethanolic leaves extract on uterus of pregnant mice and its teratogenicity

Moringa oleifera Lam (Moringaceae) is a source of additional dietary. It contains 9 essential amino acids, calcium, iron, potassium, magnesium, zinc and vitamins A, C, E and B which have a major role in the immune system. Previous study reported that M. oleifera Lam can increase the hemoglobin of breastfeeding mothers. In contrast, M. oleifera Lam is well known as abortifacient plant, therefore it is not recommended to use during pregnancy. The aim of this study was to evaluate the effect of M. oleifera Lam ethanolic leaves extract M. oleifera Lam on uterus of pregnant mice and its teratogenic effect. Twenty pregnant female Balb/c mice were used in this experimental study. Mice were randomized divided into four groups with five mice in each group. Group C as control, mice were not given anything. Group T1-T3 were given M. oleifera Lam ethanolic leaves extract during pregnancy age 7-18 days with doses of 10, 20 and 30 mg/kg BW, respectively. Phytochemical analysis was performed quantitatively and qualitatively. On day 19, a caesarean section was performed to take the uterus, count the number of living foetus, dead foetus, disability, and other morphological abnormalities. No significant difference between all groups was observed. The abortifacient effect was not also observed in the uterus. Qualitative phytochemical analysis results showed that alkaloids, flavonoids, phenols, steroids, and tannins are identified in this extract. Quantitative steroids analysis results showed that the extract contained steroids of 1057.6 ppm. In conclusion, the M. oleifera Lam ethanolic leaves extract at dose of 30 mg/kg BW does not show any effect on the uterus of pregnant mice. Further study concerning the effects on other organs and hormones is recommended.

A Highly Sensitive Modified Glassy Carbon Electrode with a Carboxylated Multi-walled Carbon Nanotubes/Nafion Nano Composite for Voltammetric Sensing of Dianabol in Biological Fluid.

The extraordinary prerequisite for the analysis of an anabolic steroid, namely dianabol (DB), has inspired towards the development of a cost-effective and high-performance sensing probe. Thus, a simple and robust electrochemical sensor (c-MWCNTs-Nafion®lGCE) for dianabol (DB), a widely used steroid, was developed using a glassy carbon electrode (GCE) modified with functionalized carboxylated multi-walled carbon nanotubes (c-MWCNT) and Nafion®. At pH 7 - 8, differential pulse-cathodic stripping voltammetry (DP-CSV) displayed two cathodic peaks at -0.85 and -1.35 V that varied linearly over a wide range (9.0 × 10-9 (2.7 μg L-1) - 9.0 × 10-6 (2.7 × 103 μg L-1) mol L-1) and 2.9 × 10-6 (8.7 × 102 μg L-1) - 8.0 × 10-5 (2.4 × 104 μg L-1) mol L-1) of DB concentrations, respectively. The low limits of detection and quantification at peak I (-0.85 V) were 2.7 × 10-9 (8.1 × 10-1 ng mL-1) and 9.0 × 10-9 (2.7 ng mL-1) mol L-1, respectively. The repeatability and reproducibility displayed relative standard deviations lower than 5%. The method was applied for DB analysis in human urine and subsequently compared with the standard HPLC method. Interference of common metabolites in biological fluids samples to DB sensing was insignificant. This method has distinctive advantages e.g. precise, short analytical time, sensitive, economical, reproducible and miniaturized sample preparation for DB analysis in biological samples of human origin.

Overcoming matrix effects in quantitative liquid chromatography-mass spectrometry analysis of steroid hormones in surface waters.

Accurate and reliable measurements are mandatory in the field of environmental monitoring. Matrix effects are often depicted as the Achilles' heel of liquid chromatography-mass spectrometry analysis since they may be prejudicial for analytical performances such as detection capability and accuracy, if not documented or compensated. Here a methodology for the evaluation and compensation of matrix effects is described. Natural and synthetic representative water samples were used for the evaluation of matrix effects with the post-extraction addition technique. Samples were analysed using ultra-performance liquid chromatography separation coupled to tandem mass spectrometry and electrospray ionization. Isotopic dilution was investigated as a way to allow compensation of signal alteration and therefore satisfactory quantification. When this approach was not possible, a methodology was conducted for choosing the most appropriate internal standard. The matrix effects were dependent on both matrix composition and nature of analyte. They ranged from total signal suppression to signal enhancement of +27% but were independent of compound concentration. The correction of matrix effects by internal standards was satisfactory, particularly for compounds benefiting from isotope dilution leading to acceptable quantification performances. Even if no exhaustive or agreed criteria exist for the final interpretation of matrix effects, this study highlights the interest in isotope dilution for reducing their inherent prejudicial effects in quantification and the need to conduct this type of study for representative matrices. Moreover, a methodological approach is proposed for choosing the most appropriate available internal standard when isotope dilution is not possible.

Clinical applications of genetic analysis and liquid chromatography tandem-mass spectrometry in rare types of congenital adrenal hyperplasia

BackgroundOur study aims to summarize the clinical characteristics of rare types of congenital adrenal hyperplasia (CAH) other than 21-hydroxylase deficiency (21-OHD), and to explore the clinical applications of genetic analysis and liquid chromatography tandem-mass spectrometry (LC-MS/MS) in rare CAH.MethodsWe retrospectively analysed the clinical data of 5 rare cases of CAH admitted to our hospital and summarized their clinical manifestations, auxiliary examinations, diagnosis and mutational spectrum.ResultsAfter gene sequencing, complex heterozygous variants were detected in all patients (2 cases were lipoid congenital adrenal hyperplasia (LCAH), 11β-hydroxylase deficiency (11β-OHD), 3β-hydroxysteroid dehydrogenase deficiency (3β-HSD deficiency) and P450 oxidoreductase deficiency (PORD) each accounted for 1 case), which were consistent with their clinical manifestations. Among them, 4 novel variants were detected, including c.650 + 2 T > A of the StAR gene, c.1145 T > C (p. L382P) of the CYP11B1 gene, c.1622C > T (p. A541V) and c.1804C > T (p. Q602 *) of the POR gene. The LC-MS/MS results for steroid hormones in patients were also consistent with their genetic variants: 2 patients with LCAH showed a decrease in all steroid hormones; 11β-OHD patient showed a significant increase in 11-deoxycortisol and 11-deoxycorticosterone; patient with 3β-HSD deficiency showed a significant increase in DHEA; and PORD patient was mainly characterized by elevated 17OHP, progesterone and impaired synthesis of androgen levels.ConclusionsThe clinical manifestations and classification of CAH are complicated, and there are cases of missed diagnosis or misdiagnosis. It’s necessary to combine the analysis of clinical manifestations and auxiliary examinations for diagnosis; if necessary, LC-MS/MS analysis of steroid hormones or gene sequencing is recommended for confirming diagnosis and typing.

Solvent Effects in the Crystalline Sponge Method: Importance of Co-solvents for Ordering Absorbed Guests.

In the crystalline sponge method, the crucial step for ordering the absorbed guest is soaking of the guest into the pores of the crystalline sponge. Here, we find that the choice of solvent is particularly important for smooth guest soaking and ordering. Moderately polar solvents, such as ketones and esters, which we have previously avoided for the guest-soaking process, efficiently promote diffusion and guest ordering by filling the gaps in the pores through co-crystallization with the guests. Using this modified protocol, we successfully demonstrate the structural analysis of various steroids.

Dexamethasone 12mg versus 6mg for patients with COVID-19 and severe hypoxaemia: a pre-planned, secondary Bayesian analysis of the COVID STEROID 2 trial.

PurposeWe compared dexamethasone 12 versus 6 mg daily for up to 10 days in patients with coronavirus disease 2019 (COVID-19) and severe hypoxaemia in the international, randomised, blinded COVID STEROID 2 trial. In the primary, conventional analyses, the predefined statistical significance thresholds were not reached. We conducted a pre-planned Bayesian analysis to facilitate probabilistic interpretation.MethodsWe analysed outcome data within 90 days in the intention-to-treat population (data available in 967 to 982 patients) using Bayesian models with various sensitivity analyses. Results are presented as median posterior probabilities with 95% credible intervals (CrIs) and probabilities of different effect sizes with 12 mg dexamethasone.ResultsThe adjusted mean difference on days alive without life support at day 28 (primary outcome) was 1.3 days (95% CrI −0.3 to 2.9; 94.2% probability of benefit). Adjusted relative risks and probabilities of benefit on serious adverse reactions was 0.85 (0.63 to 1.16; 84.1%) and on mortality 0.87 (0.73 to 1.03; 94.8%) at day 28 and 0.88 (0.75 to 1.02; 95.1%) at day 90. Probabilities of benefit on days alive without life support and days alive out of hospital at day 90 were 85 and 95.7%, respectively. Results were largely consistent across sensitivity analyses, with relatively low probabilities of clinically important harm with 12 mg on all outcomes in all analyses.ConclusionWe found high probabilities of benefit and low probabilities of clinically important harm with dexamethasone 12 mg versus 6 mg daily in patients with COVID-19 and severe hypoxaemia on all outcomes up to 90 days.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00134-021-06573-1.

A developed HPLC-MS/MS method to quantitate 5 steriod hormones in clinical human serum by using PBS as the surrogate matrix

Steroid hormones play an essential role in regulating physiological and reproductive development throughout the lifetime of an individual. One of the difficulties in determining endogenous substances is the lack of a blank matrix. Especially when the level of analytes is lower than the level in the so-called blank matrix. In the present study, an optimized HPLC-MS/MS method was developed and validated to quantify androstenedione (ASD), testosterone (Ts), dehydroepiandrosterone (DHEA), 5α-dihydrotestosterone (DHT), and progesterone (P) in serum samples from healthy people using PBS (pH = 7.4) as the blank surrogate matrix. Simultaneously, the method investigated the characteristics of NaCl, bull serum albumin, pure water as surrogate matrices for the analysis of steroid hormones. The data showed that the matrix effects of ASD, Ts, DHEA, DHT, and P in the same groups were not significantly different between PBS and twice charcoal-stripped serum (CS2S) as a blank surrogate matrix. Furthermore, the LLOQ using PBS as the blank matrix was up to 0.005 ng/mL for ASD, Ts, and P and 0.05 ng/mL for DHEA and DHT. The reference ranges of concentration (CPBS) of 5 steroid hormones were provided. Compared to the concentration with CS2S (CCSS) as the blank surrogate matrix, the relative biases (RBs) of Ts, DHT, P, and DHEA were finally stabilized at approximately −0.7%, −15%, −1.2%, and 9.2%, respectively. The results suggest that, in the cases of special required, the developed HPLC-MS/MS method can be used to determine the absolute concentration of 5 hormones in biological samples with PBS as the blank surrogate matrix.

Systems analysis of steroid induced osteonecrosis shows role for heme and vitamin D in pathogenesis

ObjectiveFemoral Head Osteonecrosis (FHO) which includes Steroid-induced FHO (SFHO) is a debilitating disease. Though extensively studied the mechanisms have remained elusive. Hence systems analysis is carried out to identify a potential mechanism, biomarkers and therapeutic targets. DesignGene Set Enrichment Analysis (GSEA) and clustering analysis were performed on GEO data sets. A Cofactor-protein interaction network was generated and pathway analysis was performed. Followed by the generation of an integrated network model. Metabolic data were analyzed using MetaboAnalyst. ADMET analysis of steroids was carried out. ResultsSystems analyses based on GSEA, Pathway clustering and Cofactor gene network showed that the enriched pathways namely Steroid, Cholesterol, Vitamin D and heme metabolism could be regulated by Vitamin B6, B12, and heme cofactors. The integrated network model showed CYP450 genes which are critical for bioactivation of Vitamin D and clearing of xenobiotic steroid metabolites which are implicated in SFHO. Analysis of transcription factors and upstream kinome showed a role for HNF4A, SREBP and GSK3b in the pathogenesis. ADMET analysis identified CYP3A4 in SFHO associated steroid clearance. Vitamin D modulated genes CBS and CYP3A4 are downregulated in SFHO potentially due to deregulated vitamin D metabolism. GSEA analysis of steroid-treated osteoblasts and cartilage show enrichment of adipogenesis, cholesterol, xenobiotic metabolism, etc. ConclusionTaken together our analysis reveals the molecular signatures associated with the disease and identifies a critical role for vitamin D, B6, B12 and heme metabolism. Low B6, B12 modulate homocysteine levels and osteoclastogenesis, while compromised heme metabolism modulate CYP450s mediated steroid clearance, Vitamin D bioactivation and influence osteoblastogenesis. Thus steroids orchestrate the events that might influence disease pathogenesis by modulating bone remodeling and thrombosis. Furthermore, our work has implications for the protection of COVID-19 patients from osteonecrosis who underwent steroid therapy.

A distinct oil group in the Dongying Depression, Bohai Bay Basin, China: New insights from norcholestane and triaromatic steroid analyses

The lower third and upper fourth members of the Eocene Shahejie Formation (lower Es3 and upper Es4) are known as two sets of effective source rocks in the Dongying Depression, Bohai Bay Basin. In this study, the saturated and aromatic steroids of 20 crude oil samples from the Dongying Depression were systematically investigated using gas chromatography–mass spectrometry (GC–MS) and GC–MS metastable reaction monitoring. Based on the distribution patterns of the steranes, norcholestanes, and triaromatic steroids, the Group I and II oils were clearly identified and shown to be derived from the lower Es3 and upper Es4 source rocks, respectively. Significantly, it was discovered that three oil samples collected from the Eocene Kongdian Formation and the Ordovician reservoirs exhibit completely different steroid and isotopic compositions from those of the Group I and II oils. These oils, as a distinct oil group (Group III), have the characteristics of abnormally high concentrations of C29 steranes, 24-nordiacholestane, 27-nordiacholestane, C28 triaromatic steroids, C27 4α-methyltriaromatic steroids, and C29 4α-methyl-24-ethyltriaromatic steroids, and low concentrations of 24-norcholestane, 27-norcholestane and triaromatic dinosteroids, and 13C depletion. Although the related source rock of Group III oils needs to be further clarified in the study area, the 13C depletion, abundant C29 steranes, and low 24-norcholestane content may suggest greater bacteria/algae inputs (e.g., green algae rather than dinoflagellates) than both the lower Es3 and upper Es4 source rocks. Identification of the Group III oils indicates that the norcholestanes and triaromatic steroids may be more effective molecular fossils for the classification of crude oils in complicated petroleum exploration fields.

Steroid hormone profiles and body conditions of migrating male humpback whales (Megaptera novaeangliae)

Simultaneous analysis of multiple steroid hormones from remotely obtained blubber biopsies has the potential to concurrently provide information regarding stress and reproductive status from free-swimming cetaceans, while also investigating correlations between hormone concentrations and other health biomarkers. In this study we measured blubber concentration profiles of eight reproductive and adrenal steroid hormones (17α-hydroxy-progesterone, testosterone, androstenedione, progesterone, cortisol, 11-deoxy-corticosterone, oestrone, and oestradiol) together with body condition, as determined by the inverse Adipocyte Index, of 101 male humpback whales. Whales were sampled randomly at two time points, while migrating to and from their northeast Australian breeding grounds, allowing for intra- and inter-seasonal profile analysis. Testosterone, progesterone and cortisol together with androstenedione 17α-hydroxyprogesterone, and oestrone concentrations (the latter quantified for the first time in live biopsied male humpback whales) decreased between the northward and southward migrations. Decreasing testosterone levels during the height of humpback whale conceptions suggests asynchronicity between blubber testosterone levels and the expected peak of male fertility. Statistically significant relationships between levels of certain steroid analytes were observed and appeared to change between the early and late breeding seasons. During the northward migration, testosterone, progesterone, androstenedione, oestrone and 17α-hydroxyprogesterone levels were positively correlated. Cortisol concentrations correlated positively with those of testosterone during the northward migration, but negatively during the southward migration. Androstenedione and testosterone were positively correlated with adiposity during the late breeding season. These hormone-hormone and hormone-adiposity correlations may be reflective of the activation of certain steroid hormone synthesis pathways, or alternatively, of concomitant physiological stimuli. As steroid hormones work in concert, information on multiple steroid hormones is needed to interpret endocrinological status and understand the relationships between these compounds and ancillary health markers. This study provides steroid hormone profiles of wild male humpback whales, as well as the first insight into seasonal male endocrinology as a function of adiposity.

Multidimensional Separations of Intact Phase II Steroid Metabolites Utilizing LC–Ion Mobility–HRMS

The detection and unambiguous identification of anabolic-androgenic steroid metabolites are essential in clinical, forensic, and antidoping analyses. Recently, sulfate phase II steroid metabolites have received increased attention in steroid metabolism and drug testing. In large part, this is because phase II steroid metabolites are excreted for an extended time, making them a potential long-term chemical marker of choice for tracking steroid misuse in sports. Comprehensive analytical methods, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), have been used to detect and identify glucuronide and sulfate steroids in human urine with high sensitivity and reliability. However, LC-MS/MS identification strategies can be hindered by the fact that phase II steroid metabolites generate nonselective ion fragments across the different metabolite markers, limiting the confidence in metabolite identifications that rely on exact mass measurement and MS/MS information. Additionally, liquid chromatography-high-resolution mass spectrometry (LC-HRMS) is sometimes insufficient at fully resolving the analyte peaks from the sample matrix (commonly urine) chemical noise, further complicating accurate identification efforts. Therefore, we developed a liquid chromatography-ion mobility-high resolution mass spectrometry (LC-IM-HRMS) method to increase the peak capacity and utilize the IM-derived collision cross section (CCS) values as an additional molecular descriptor for increased selectivity and to improve identifications of intact steroid analyses at low concentrations.

Simultaneous Analysis of Sexual Stimulants and Anabolic Steroids as Adulterants in Dietary Supplements by High Performance Liquid Chromatography with Photodiode Array Detection

Background:The illegal virtual market for food supplements facilitates fraud and adulteration. Worldwide concern for consumer safety is growing on the part regulatory agencies, healthcare professionals and consumers.Objective:This work aimed to evaluate the presence of sexual stimulants and anabolic steroids commonly used in the adulteration of dietary supplements through the development of a high performance liquid chromatography with photodiode array detection (HPLC-DAD) method for the identification and quantification of these compounds.Method:The mobile phase composed of an ammonium acetate solution, acetonitrile and methanol leaded to the efficient separation of vardenafil, testosterone base, testosterone propionate, tadalafil, sildenafil and yohimbine.Results:The assay was linear (r2&gt; 0.999), precise (RSD% &lt;0.5), accurate (99.1 to 105.2%), and the limits of detection and quantification were less than 0.05 and 0.15 μg/mL, respectively. Four samples of dietary supplements contained testosterone (n=1), tadalafil (n=2) and yohimbine (n=1) as adulterants. The adulterants found were in subtherapeutic doses, probably to reduce possible adverse effects and the action expected to appear natural. Since about 80% of adverse drug reactions are dose dependent, unpredictable adverse drug reactions are dose independent and based on idiosyncratic or allergic mechanisms or intolerance.Conclusion:The developed method is convenient and easily applicable for adulteration detection of the analyzed drugs in the multicomponent supplements.