A method for determination of one hundred endogenous steroids in human serum by gas chromatography-tandem mass spectrometry.

Abstract

Steroid profiling helps various pathologies to be rapidly diagnosed. Results from analyses investigating steroidogenic pathways may be used as a tool for uncovering pathology causations and proposals of new therapeutic approaches. The purpose of this study was to address still underutilized application of the advanced GC-MS/MS platform for the multicomponent quantification of endogenous steroids. We developed and validated a GC-MS/MS method for the quantification of 58 unconjugated steroids and 42 polar conjugates of steroids (after hydrolysis) in human blood. The present method was validated not only for blood of men and non-pregnant women but also for blood of pregnant women and for mixed umbilical cord blood. The spectrum of analytes includes common hormones operating via nuclear receptors as well as other bioactive substances like immunomodulatory and neuroactive steroids. Our present results are comparable with those from our previously published GC-MS method as well as the results of others. The present method was extended for corticoids and 17α-hydroxylated 5α/β-reduced pregnanes, which are useful for the investigation of alternative "backdoor" pathway. When comparing the analytical characteristics of the present and previous method, the first exhibit by far higher selectivity, and generally higher sensitivity and better precision particularly for 17α-hydroxysteroids. Key wordsSteroid metabolome • Human blood • Gas chromatographytandem mass spectrometry • Backdoor pathway • Pregnancy • Mixed umbilical cord blood Methods SamplesSerum samples from non-pregnant subjects were collected from the employees of the Institute of Endocrinology, Prague, Czech Republic and their relatives, as well as from patients of the Institute of Endocrinology. Serum samples from pregnant women and umbilical cord serum at birth were obtained from patients

Alteration of the steroidogenesis in boys with autism spectrum disorders

“…Unfortunately, some of the major hormonal steroids being the important intermediates and products like 5α-dihydrotestosterone, which would have emphasized our findings, were not detected. Unfortunately, we did not detected mineralocorticoids like aldosterone using the GC-MS/MS due to the chemical nature of these molecules 21 . Another limitation might be that control children in the control group did not undergo the diagnostic procedure ADOS-2 and ADI-R.…”

“…All steroids and their polar conjugates were analyzed using gas-chromatography tandem-mass spectrometry (GC-MS/MS). Whole procedure was performed according to Hill et al 21 .…”

Activation of Adrenal Steroidogenesis and an Improvement of Mood Balance in Postmenopausal Females after Spa Treatment Based on Physical Activity

“…The following circulating steroids were quantified by gas chromatography tandem mass spectrometry (GC-MS/MS) (n = 88) using our recently published method [23]: Pregnenolone, pregnenolone sulfate, 17-hydroxypregnenolone, 17-hydroxypregnenolone sulfate, 16α-hydroxypregnenolone, 20αdihydropregnenolone, 20α-dihydropregnenolone sulfate, dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), 7α-hydroxy-DHEA, 7-oxo-DHEA, 7β-hydroxy-DHEA, androstenediol, androstenediol sulfate, 5-androstene-3β,7α,17β-triol, 5-androstene-3β,7β,17β-triol, 5-androstene-3β,16α,17β-triol, 5-androstene-3β,16α,17β-triol sulfate, progesterone, 17-hydroxyprogesterone, 17α,20α-dihydroxy-4-pregnen-3-one, 16αhydroxyprogesterone, 20α-dihydroprogesterone, conjugated 20α-dihydroprogesterone, androstenedione, testosterone, conjugated testosterone, 5α-dihydrotestosterone, estradiol, conjugated estradiol, 5αdihydroprogesterone, allopregnanolone, allopregnanolone sulfate, isopregnanolone, isopregnanolone sulfate, pregnanolone, conjugated pregnanolone, epipregnanolone, conjugated epipregnanolone, 5α,20α-tetrahydroprogesterone, conjugated 5α,20α-tetrahydroprogesterone, 5α-pregnane-3α,20α-diol, conjugated 5α-pregnane-3α,20α-diol, 5α-pregnane-3β,20α-diol, conjugated 5α-pregnane-3β,20α-diol, 5β,20α-tetrahydroprogesterone, conjugated 5β,20α-tetrahydroprogesterone, 5β-pregnane-3α,20α-diol, conjugated 5β-pregnane-3α,20α-diol, 5β-pregnane-3β,20α-diol, conjugated 5β-pregnane-3β,20α-diol, 17-hydroxyallopregnanolone, 17-hydroxyallopregnanolone sulfate, 17-hydroxypregnanolone, 17hydroxypregnanolone sulfate, 5α-pregnane-3α,17α,20α-triol, conjugated 5α-pregnane-3α,17α,20α-triol, 5α-pregnane-3β,17α,20α-triol, conjugated 5α-pregnane-3β,17α,20α-triol, 5β-pregnane-3α,17α-20α-triol, conjugated 5β-pregnane-3α,17α,20α-triol, 5α-androstane-3,17-dione, androsterone, androsterone sulfate, epiandrosterone, epiandrosterone sulfate, etiocholanolone, etiocholanolone sulfate, epietiocholanolone, epietiocholanolone sulfate, 5α-androstane-3α,17β-diol, conjugated 5α-androstane-3α,17β-diol, 5αandrostane-3β,17β-diol, conjugated 5α-androstane-3β,17β-diol, 5β-androstane-3α,17β-diol, conjugated 5β-androstane-3α,17β-diol, cortisol, cortisone, corticosterone, 5α,20α-tetrahydrocorticosterone, 5β,20αtetrahydrocorticosterone, 11β-hydroxyandrostenedione, 11β-hydroxyandrosterone, 11β-hydroxyandrosterone sulfate, 11β-hydroxyepiandrosterone, 11β-hydroxyepiandrosterone sulfate, 11β-hydroxyetiocholanolone, 11β-hydroxyetiocholanolone sulfate.…”

“…Sleep disturbance; 2. daytime sleepiness; 3. fatigue and exhaustion; 4. low productivity; 5. headache; 6. perspiration; 7. vertigo; 8. faint feeling; 9. vomiting and nausea after anger; 10. hot and cold flashes; 11. shivers, inner need; 12. inattention and lack of concentration; 13. irritability and short temper; 14. memory retention; 15. backbone, muscle, and joint pain; 16. heart palpitations, heart pain; The intensity of the symptoms was self-estimated by patients using a 4-level scale: 0 = no, 1 = mild, 2 = high, 3 = extreme. The total score (from the viewpoint of neurotic tendency) was interpreted as follows: Normal (0-11), Mildly neurotic (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22) Highly neurotic (23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33), Extremely neurotic (>34). The somatic and psychosomatic disturbances were reflected in questions 1-11 and 12-22, respectively, while psychiatric symptoms were represented by questions 23-33.…”

The Potential of Steroid Profiling by Mass Spectrometry in the Management of Adrenocortical Carcinoma

“…The biological fluids most used in clinical practice for the determination of steroids in GC-MS are: serum [47][48][49], plasma [50], and urine [51,52].…”