Important considerations for the utilisation of methanolysis in steroid analysis

2018 ◽  
Vol 10 (9) ◽  
pp. 1469-1473 ◽  
Author(s):  
Marjaana Viljanto ◽  
Catarina Horro Pita ◽  
James Scarth ◽  
Christopher J. Walker ◽  
Andrew T. Kicman ◽  
...  
Keyword(s):  
Steroid Analysis

Isolated 17, 20 Lyase Deficiency Secondary to a Novel CYB5A Variant: Comparison of Steroid Metabolomic Findings with Published Cases Provides Diagnostic Guidelines and Greater Insight into Its Biological Role

2020 ◽  
Vol 93 (7-8) ◽  
pp. 483-496
Author(s):  
Meera Shaunak ◽  
Norman F. Taylor ◽  
David Hunt ◽  
Justin H. Davies
Keyword(s):  
Mass Spectrometry ◽  
Ambiguous Genitalia ◽  
Relative Increase ◽  
Testicular Microlithiasis ◽  
Lyase Deficiency ◽  
Chain Cleavage ◽  
Steroid Profiles ◽  
Steroid Analysis ◽  
17 Hydroxyprogesterone ◽  
Urine Steroid

Objective: The objective of this study was to report CYB5A deficiency, to discuss the contribution of steroid metabolomics to diagnosis and interpretation, and to highlight the presence of testicular microlithiasis. Methods: Two siblings with ambiguous genitalia at birth were later found to carry novel CYB5A variants, with resulting isolated 17, 20 lyase deficiency. We compared urine steroid data obtained between birth and adulthood with that from other cases. Results: Neonatal urine steroid profiles show a relative increase of 16-hydroxylated pregnenolone metabolites. Thereafter, there are no distinguishing features until puberty, when sex steroid deficiency drives gonadotrophin production, resulting in marked increases of 17-hydroxyprogesterone metabolites derived from the gonads. This excess may be revealed pre-pubertally by gonadotrophin stimulation testing. Novel findings are first, a considerable capacity for DHEA synthesis in the neonatal period compared to childhood and adulthood, suggesting that DHEAS production is much less dependent on CYB5A at birth; second, no consistent change in “backdoor pathway” intermediates; third, side chain cleavage of cortisol is largely unaffected, supporting the existence of a different lyase not dependent on CYB5A; fourth, increased 17-hydroxyprogesterone metabolites and very low androgen metabolites are diagnostic post-pubertally. Conclusion: This is the fourth disease-causing variant in CYB5A in isolated 17, 20 lyase deficiency and the first associated with testicular microlithiasis. Establishing a biochemical diagnosis pre-pubertally should now be possible using urine steroid profiling, supported by synacthen and gonadotrophin stimulation testing. We recommend liquid chromatography-mass spectrometry/mass spectrometry rather than immunoassay for serum steroid analysis, early methaemoglobin measurement and surveillance should testicular microlithiasis be detected.

scholarly journals Increased speed of steroid analysis by capillary GC and GC-MS; effects of sample pretreatment and sample introdution

1984 ◽  
Vol 7 (7) ◽  
pp. 414-422 ◽  
Author(s):  
J. Curvers ◽  
F. Maris ◽  
C. Cramers ◽  
C. Schutjes ◽  
J. Rijks
Keyword(s):  
Sample Pretreatment ◽  
Capillary Gc ◽  
Steroid Analysis

USE OF ION EXCHANGE CHROMATOGRAPHY IN STEROID ANALYSIS

1983 ◽  
pp. 175-180
Author(s):  
RISTO HEIKKINEN ◽  
THEODORE FOTSIS ◽  
HERMAN ADLERCREUTZ
Keyword(s):  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Steroid Analysis

scholarly journals Steroid analysis by ion mobility spectrometry

Steroids ◽  
2020 ◽  
Vol 153 ◽  
pp. 108531 ◽  
Author(s):  
Alana L. Rister ◽  
Eric D. Dodds
Keyword(s):  
Ion Mobility Spectrometry ◽  
Ion Mobility ◽  
Steroid Analysis

100 Reproductive cycle and pregnancy monitoring in the common hippopotamus (Hippopotamus amphibius) through salivary steroid analyses and transabdominal ultrasonography

2019 ◽  
Vol 31 (1) ◽  
pp. 176
Author(s):  
J. Wojtusik ◽  
I. M. C. Brandicourt ◽  
W. Rice ◽  
T. L. Roth
Keyword(s):  
Enzyme Immunoassay ◽  
Reproductive Cycle ◽  
Reproductive Activity ◽  
Botanical Garden ◽  
Ex Situ ◽  
Sample Collection ◽  
Transabdominal Ultrasonography ◽  
Hippopotamus Amphibius ◽  
Steroid Analysis ◽  
The Common

The common hippopotamus (Hippopotamus amphibius) is listed as vulnerable to extinction by the IUCN due to a significant decrease in population size, caused by habitat loss and poaching. Ex situ populations can help ensure against species loss, but careful reproductive management is essential to maintain sustainable populations. Hormone monitoring allows for characterisation of the reproductive cycle and gestation, offering insight into timing of receptivity and conception and facilitating pregnancy diagnosis and estimation of parturition date. Fecal steroid analysis has been validated for measuring progestogens in hippos. However, hippos are often housed in groups and frequently defecate in the water, making sample collection and source identification difficult. Salivary steroid analysis has been employed for monitoring reproductive activity in several species, but has not been tested in hippos. Additionally, transabdominal ultrasonography has proven valuable in diagnosing and monitoring pregnancy in many large mammals, but efficacy in the common hippo is unknown. The goals of this project were to (1) validate the use of an enzyme immunoassay to monitor progestogens in hippo saliva, (2) confirm that salivary progestogen profiles accurately reflect reproductive activity, (3) determine if transabdominal ultrasonography can be used to diagnose pregnancy, and, if so, (4) monitor and characterise fetal development via weekly examinations. Saliva (4-7 per week) and fecal (2-7 per week) samples were collected from 7 adult female hippos housed at 3 USA facilities over 3-7 months. Saliva and fecal samples were extracted in ethanol and extracts diluted (1:2 to 1:10 and 1:25 to 1:500, respectively) before evaluation by enzyme immunoassay (Progesterone mini-kit; Arbor Assays). Parallelism was confirmed between serially diluted fecal (r2=0.993) and saliva (r2=0.990) samples and the standard curve. Inter- and intra-assay coefficients of variation were maintained at <10%. Comparison of fecal and saliva progestogen concentrations revealed a strong correlation between the 2 sample types (r2=0.848) and suggested that saliva offers a comparable alternative. Both fecal and saliva extracts exhibited elevated progestogens during luteal phases and gestation. One nulliparous female housed at the Cincinnati Zoo & Botanical Garden (Cincinnati, OH, USA) was trained for voluntary transabdominal ultrasound exams. An Ibex Pro portable ultrasound machine (E.I. Medical Imaging, Loveland, CO, USA) with curvilinear probe (5-2.5MHz) was used at a scanning depth of 17.8 and 23.4cm. Intrauterine fluid and possible fetal tissue were observed 79 days following the last confirmed mating. Spine, rib cage, and beating heart were clearly visible at ~156 days of gestation. Ultrasound procedures were continued until the premature birth of a calf at ~181 days (normal hippo gestation ~231 days). Salivary progestogen monitoring and transabdominal ultrasonography appear suitable for tracking reproductive activity and diagnosing and monitoring pregnancy in the common hippo.

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