Steroid compounds analysis

Many methods have been used to quantify steroidal compounds. These include RIA, gas chromatogra-phy-mass spectrometry (GC/MS), high-performance liquid chromatography (HPLC), and liquid chroma-tography-mass spectrometry (LC/MS). Although these techniques are successful in the analysis of steroids, it has been difficult to achieve quantitative analysis of small samples of neurosteroids because of their low concentrations in nervous tissues. Highly specific analytical methods are required to analyze small quantities of neurosteroids and their sulfates. Only with extremely sensitive methods of analysis is it possible to discover whether neurosteroids are synthesized in nervous tissues in quantities sufficient to affect neuronal activity, and whether these neurosteroids are distributed uniformly in brain. 

A. Marwah, P. Marwah, H. Lardy, Analysis of ergosteroids - VIII Enhancement of signal response of neutral steroidal compounds in LC-ESI-AIS analysis by mobile phase additives, J. Chromatogr. A, 964 (2002) 137. 

Steroidal compounds have been shown to act as inclusion hosts. The Miyata group has made the development and interpretation of this chemistry very much their own through their skillful analysis of over 300 inclusion crystal structures. Concave face to concave face association by means of hydrogen bonding results in bilayer formation in many of these inclusion crystals, but a very wide range of different host-guest structural assembUes is possible. One crystal form of the compound (4) (ethyl acetate) is illustrated in Figure 2 ... 

The thermospray interface was the first LC-MS system that allowed reliable quantitative bioanalysis for a wide variety of compounds. Numerous examples were published in the literature. An excellent example is the automated analysis of bambuterol. The automated system, described by Lindberg and coworkers, contained a series of feedback steps in order to assure the various components of the system were operating properly during overnight, unattended analysis and to avoid the loss of valuable sample material. The same approach was applied to the quantitative bioanalysis of cortisol and related steroid compounds. In order to enhance the response of cortisol in thermospray ionization, the compound was derivatized to the 21-acetate using acetic... 

Residual aromatic ether concentrations are determined from the absorbance at 278 mfi of the crude reduction products in methanol solution. Steroidal ether concentrations of 1 mg/ml are employed. The content of 1,4-dihydro compound is determined, when possible, by hydrolysis to the a, -unsaturated ketone followed by ultraviolet analysis. A solution of the crude reaction product (usually 0.01 mg/ml cone) in methanol containing about 1/15 its volume of water and concentrated hydrochloric acid respectively is kept at room temperature for 2 to 4 hr. The absorbance at ca. 240 mfi is measured and, from this, the content of 1,4-dihydro compound can be calculated. Longer hydrolysis times do not increase the absorbance at 240 mp.. 

Pentafluorobenzyl bromide has been used in the derivatization of mercaptans [55] and phenols [36], m the analysis of prostaglandins [37], and in quantitative GC-MS [5S] 1,3 Dichlorotetrafluoroacetone is used for the derivatization of amino acids to the corresponding cyclic oxazolidinones and allows the rapid analysis of all 20 protein ammo acids [d] Pentafluorophenyldialkylchlorosilane derivatives have facilitated the gas chromatographic analysis of a wide range of functionally substituted organic compounds, including steroids, alcohols, phenols, amines, carboxylic acids, and chlorohydrms [4]... 

Polycyclic compounds are common in nature, and many valuable substances have fused-ring structures. For example, steroids, such as the male hormone testosterone, have 3 six-membered rings and 1 five-membered ring fused together. Although steroids look complicated compared with cyclohexane or decalin, the same principles that apply to the conformational analysis of simple cyclohexane lings apply equally well (and often better) to steroids. 

Applications APCI-MS is often more widely applicable than ESI-MS to the analysis of classes of compounds with a low molecular weight, such as basic drugs and their metabolites, antibiotics, steroids, oestrogens, benzodiazepines, pesticides, surfactants, and most other organic compounds amenable to El. LC-APCI-MS has been used to analyse PET extracts obtained by a disso-lution/precipitation procedure [147]. Other applications of hyphenated APCI mass spectrometric techniques are described elsewhere LC-APCI-MS (Section 7.33.2) and packed column SFC-APCI-MS (Section 73.2.2) for polar nonvolatile organics. 

The analysis of steroid sexual hormones and related synthetic compounds in WW, soil, sludge, and sediment samples is a challenging task. This is due to both the complex environmental matrices and the requirement of low detection limits. Therefore, the use of complicated, time- and labor-consuming analytical procedures is necessary. 

In total, 185 substances were found in the wing-sac liquid of male S. bilineata from a Costa Rican population. For a more detailed analysis the relative peak area of thirteen focus compounds was compared (Table 14.1). Of these nine were male-specific substances (indole, indol-3-carboxaldehyde, indole-3-carboxylic acid, 2-aminoacetophenon, anthranilic acid, SHJOH-dipyrrolofl -aT -dJpyrazine-5,10-dione (pyrocoll), indolo[2,l-b]quinazoline-6,12-dione (tryptanthrin), 2,6,10-trimethyl-3-oxo-6,10-dodecadienolide, and a compound C15H24O2 of unknown structure), three were fatty acids (tetradecanoic acid, hexadecanoic acid, and octadecanoic acid), and one a steroid (cholesterol). On average, the cumulative peak area of these substances made up 62.5 20.7% of the whole chromatogram area. 

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