Quantification of steroids

Kolodzieg E.P., J.L. Gray, and D.L. Sedlak (2000). Quantification of steroid hormones with phenomenal properties in murucipal wastewater effluent. Environmental Toxicology and Chemistry 22 2622-2629. 

Vol. XXXrV [1], The Literature on Affinity Chromatography. M. Wilchek and W. B. Jakoby. Vol. XXXVI [1]. Theory of Protein-Ligand Interaction. D. Robard and H. A. Feldman. Vol. XXXVI [2]. Use of Specific Antibodies for Quantification of Steroid Hormones, G. D. Niswender, A. M. Akbar, and T. M. Nett. 

Quignot N, Tournier M, Pouech C, Cren-Olive C, Barouki R, Lemazurier E (2012) Quantification of steroids and endocrine disrupting chemicals in rat ovaries by LC-MS/MS for reproductive toxicology assessment. Anal Bioanal Chem 403 1629-1640. doi 10.1007/ s00216-012-5990-y... 

The use of isotopically labeled steroids as internal standards for the quantification of steroids in biofluids has been refined to a high degree. Four crucial steps in the analysis have been identified, namely... 

Pozo, O.J., Van Eenoo, E, Van Thuyne, W., et al (2008) Direct quantification of steroid glucuronides in human urine by liquid chromatography-electrospray tandem mass spectrometry. Journal of Chromatography A, 1183,108-118. 

A definitive method for stmctural deterrnination is x-ray crystallography. Extensive x-ray crystal stmcture deterrninations have been done on a wide variety of steroids and these have been collected and Hsted (270). In addition, other analytical methods for steroid quantification or stmcture determination include, mass spectrometry (271), polarography, fluorimetry, radioimmunoassay (264), and various chromatographic techniques (272). 

Nixon A., Mallet A.I., et al. (1988). Simultaneous quantification of five odorous steroids 16-Androstenes, in the axillary hair of men. J Steroid Biochem 29, 505-510. 

The routine measurement of estrone and estradiol by immunoassay techniques has also given rise to the familiar problems of poor sensitivity, cross-reactivity, and poor intermethod reproducibility. Most automated methods cannot measure these steroids in sera of children and men. Development of LC-MS/MS methods has also proven challenging as most investigators have found that estrone and estradiol are poor ionizers and the desired sensitivity has not been achieved. The Mayo group [60] published the LC-MS/MS quantification of estradiol and estrone using dan-syl chloride derivatives (described above) and 2H5 estradiol and 2H4 estrone internal standards. 

Rauh M, Groschl M, Rascher W, Dorr HG (2006) Automated, fast and sensitive quantification of 17 alpha-hydroxy-progesterone, androstenedione and testosterone by tandem mass spectrometry with on-line extraction. Steroids 71 450-458... 

McClelland, R. A., Wilson, D., Leake, R., Finlay, P., and Nicholson, R. I. 1991. A multicentre study into the reliability of steroid receptor immunocytochemical assay quantification. Eur. J. Cancer 27 711-715. 

Hala D, Overturf MD, Petersen LH, Huggett DB (2011) Quantification of 2-hydrazinopyri-dine derivatized steroid hormones in fathead minnow (pimephales promelas) blood plasma using LC-ESI+/MS/MS. J Chromatogr B 879 591-598... 

Much effort has been made to detect steroids in biological fluids. Even simple TLC methods have been used for qualitative analysis [38], One method that been used for quantification involves an immunoassay, but several problems exist with that method, most notably cross-reactions and interference with other substances [39], On the other hand, a number of chromatographic methods have been developed to overcome these problems. The majority of analytical methods involved GC, which has good detection limits, but requires previous derivatization [40] of the steroids to accomplish volatilization. Many methods have also been reported using HPLC with UV detection or LC-MS [40, 41], Previously used stationary phases for LC was e.g., Sephadex LH-20, Celite and Lipidex, but they could not be operated with high pressure [42], These columns were therefore slow to run and the separation of steroids was very time-consuming [43], Nowadays applications mainly use HPLC as a separation method with both normal-phase and re-versed-phase chromatography. 

Figure 12 Illustration of the quantification of 24-hour cortisol suppression during multiple dosing of an inhaled steroid given b.i.d. The difference between placebo, or baseline (dashed line), and active treatment (solid line) can be used to calculate the degree of cortisol suppression. (Generated using an Excel spreadsheet developed by S. Krishnaswami et al., Ref. 114.)...

Celius, T., J.B. Matthews, J.P. Giesy and T.R. Zacharewski. Quantification of rainbow trout Oncorhynchus mykiss) zona radiata and vitellogenin mRNA levels using real-time PCR after in vivo treatment with estradiol-17/3 or a-zeralenol. Steroid Biochem. Mol. Biol. 75 109-119, 2000. 

R7. Reynolds, J. W., The identification and quantification of A -androstenetriol isolated from the urine of premature infants. Steroids 8, 719-727 (1966). 

Simmer, H. H., Easterling, W. E., Jr., Pion, R. J., and Dignam, W. J., Identification of dehydroepiandrosterone sulfate in fetal blood and quantification of this hormone in cord arterial, cord venous and maternal peripheral blood in normal pregnancies at term. Steroids 4, 125-135 (1964). 

Immunoassays can be designed in two formats competitive assays, preferable for quantification of small molecules such as steroid hormones and prostaglandins, and noncompetitive, or sandwich, assays restricted almost exclusively to large molecules... 

Esteban, N.V. Yergey, A.L. Liberate, D.J. Lougfalin, T. Loriaux, D.L. Stable isotope dilution method using thermospray liquid chromatography/mass spectrometry for quantification of daily cortisol production in humans. Biomed.Environ.Mass.Spectrom., 1988, 15, 603-608 [thermospray LC-MS] Sheikh, S.U. Touchstone, J. HPLC of steroids in non-aqueous mobfle phase at subambient temperature. J.Liq.Chromatogr., 1987, 10, 2489-2496 [column temp -50 simultaneous cortisone, desoxycorticos-terone, estradiol, estrone]... 

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