A stability-indicating HPLC method for the determination of 17β-estradiol-3-phosphate in an ophthalmic solution

Summary Thep-tert-butylcalix[n]arenes (n=4,6,7,8) andp-tert-butyldihomooxacalix[4]arene have been separated by reverse phase liquid chromatography. Optimum conditions have been obtained on a Spherisorb ODS1, 5 m C18 column by isocratic ambient elution with acetonitrile-methyltert-butyl ether. Calibration plots have been obtained from purified calixarenes and the reliability of the method is confirmed from test mixtures of calixarenes of known composition.     

Synthesis of Novel 17α-Eethynyl-5-androstene-3β, 7β, 17β-triol (HE3286)Derivatives

以脱氢表雄酮为原料,经羟基乙酰化,羰基亚乙二氧基化,CrO3/3,5-二甲基吡啶氧化,硼氢化钠还原,脱17-位亚乙二氧基,脱3-位乙酰基,用叔丁基二甲基氯硅烷(TBDMSC1)保护3,7-OH,17-位环氧化,17-位开环,脱TBDMSC1保护等十步反应合成了四个新型的17α-乙炔基-5-雄甾烯-3β,7β,17β-三醇(HE3286)衍生物,其结构经1H NMR和MS表征.     

Liquid chromatography-tandem mass spectrometry analysis of 17α-trenbolone, 17β-trenbolone and trendione in airborne particulate matter

Trenbolone acetate (TbA) is a potent synthetic anabolic steroid that was approved by the FDA as a growth promoter in beef cattle in 1987. Given the endocrine-modulating activity of TbA and its metabolites in all vertebrates, a sensitive and reliable analytical method is needed to detect TbA and related residues in environmental matrices. We have developed a method that incorporates solid phase extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of the three major TbA metabolites (trendione, 17β-trenbolone, 17α-trenbolone) in total suspended particulate matter (TSP) samples. Sample preparation involved pressurized liquid extraction followed by cleanup on solid-phase extraction cartridges. The procedure was optimized to obtain maximum recovery and minimum signal suppression/enhancement from matrix effects. Analytes were separated with a Phenomenex Gemini-NX C18 analytical column (150 mm × 2.0 mm, 3 μm particle size) using an aqueous methanol gradient at a flow rate of 0.2 mL/min. Column effluent underwent positive electrospray ionization (ESI). Two or more diagnostic product ions were acquired from analyte specific precursor ions for unambiguous confirmation and quantification. The method detection limit was 3.27-4.87 ng/g of particulate matter (PM). Method accuracy, determined with analyte recoveries, ranged between 68% and 117%, and method precision, expressed as relative standard deviation, was below 15% at spiked levels of 6.67, 33.3, and 167 ng/g PM. Analysis of TSP samples demonstrated the presence of the target species associated with PM in the vicinity of beef cattle feeding operations.     

New variants in the total synthesis of 14β-hydroxy-17α,R-8α,9β-estradiol derivatives

Two procedures were developed for the synthesis of 17-alkyl-8,9-estra-1,3,5(10)-triene-3,14,17-triols from 3-methoxy-estra-1,3,5(10),8(9),14(15)-pentaene-17-one (1) using photosensitized oxidation by atmospheric oxygen or ionic hydrogenation in the presence of oxygen. It was shown that 17-hexynyl-8,9-estra-1,3,5(10)-triene-3,14,17-triol (23) and its 3-methoxy derivative (26) inhibit K⁺,Na⁺-ATPase, with inhibition degree increasing on introduction of an oxygen-containing substituent into position 17. For the first time steroids were found with ₂-adrenoblocking action.Translated fromIzvestiya Akademii Nauk, Seriya Khimicheskaya, No. 1, pp. 207–212, January, 1993.     

Synthesis of nitrogen-containing derivatives of 17(20)-pregnenoic, 17β-hydroxypregnanoic,and 17α-hydroxypregnanoic acids as new potential antiandrogens

A general scheme for the synthesis of oxazoline and benzoxazole derivatives of [17(20)E]-21-norpregnene differing in the structure of the steroid moiety as well as amides of 17β-hydroxy-3-oxopregn-4-en-21-oic and 17α-hydroxy-3-oxopregn-4-en-21-oic acids was developed. The scheme involved synthesis of the steroid building blocks (appropriately protected derivatives of pregn-17(20)-en-21-oic, 17β-hydroxypregnan-21-oic, and 17α-hydroxypregnan-21-oic acids) and subsequent transformation of these building blocks into the target compounds. Following the developed scheme, synthesis of new nitrogen-containing steroid derivatives exhibiting antiandrogenic activity was enabled.     

Laccase-mediated oxidation of the steroid hormone 17β-estradiol in organic solvents

Laccase-mediated oxidation of the steroid hormone 17β-estradiol 1 in organic solvents or in a biphasic system allowed the isolation of the C–C and C–O dimers 1a–d. Concerning the C–C dimers, the relative ratio of the symmetric 4-4′ 1c and asymmetric 4-2′ 1d products was influenced by the catalyst used. Both 1c and 1d were formed as an equimolar mixture of diastereomeric atropisomers.     

Synthesis of esters of α-chloroacetoxy-β,β,β,-trichloroethylphosphonic acids

Summary The synthesis of 11 previously unreported dialkyl esters of -mono-, -di-, and -trichloroacetoxy-,,-trichloroethylphosphonic acids has been effected.     

The structure of 16, 17β-epoxy-17α-pregn-5-en-3β-ol-20-one monohydrate

The structure of the title compound (1) has been studied by means of X-ray diffraction. The region of cycleD in epoxide1 was compared with that of the 16,17-epoxy derivatives of progesterone and pregnenolone (studied previously) as well as with that of the respective 16,17- and 16,17-cyclopropano analogs. In contrast to the 16,17-epoxy-20-oxo derivatives, in compound1 the electron conjugation of the epoxide ring with the CH₃CO group at C(17) is partly disrupted. Moreover, the steric congestion at C(17) is significantly less pronounced in 16,17-epoxide1 than in its 16,17-counterpart. Both of these factors, especially steric decongestion, are favorable for nucleophilic attack at C(17) in the molecule of1. The X-ray diffraction data do not contradict the previously advanced mechanism of epoxide ring opening in 16,17- and 16,17-epoxy-20-oxo steroids by nucleophilic reagents.Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 2, pp. 401–405, February, 1993.     

Reaction of 5β,6β-epoxy-16α,17α-cyclohexapregnane with methylmagnesium iodide

On treatment with methylmagnesium iodide, 3β-acetoxy-5β,6β-epoxy-16α,17α-cyclo-hexapregnan-20-one undergoes 3-O-deacetylation along with the opening of the 5β,6β-epoxide ring to form 5α-methyl-6β-hydroxy steroid and the 6α-methyl-6β-hydroxy isomer, the 20-keto group remaining intact.     

Effects of 17β-trenbolone in male eelpout Zoarces viviparus exposed to ethinylestradiol

To evaluate the interaction between 17β-trenbolone (TB) and 17α-ethinylestradiol (EE2), male eelpout, Zoarces viviparus, was exposed for 21 days (April to May 2008) to 5 ng l⁻¹ EE2 and 5 or 20 ng l⁻¹ TB, separately or in combination in a flow-through SW system. The effects on hepatosomatic (HSI) and gonadosomatic index (GSI), plasma vitellogenin (Vtg) concentration, gonadal histology, hepatic and testicular Vtg mRNA and estrogen receptor (ERα) mRNA expression were investigated. No effects on HSI were observed. A significant decrease was observed in the GSI of all males exposed to EE2 (<0.7%) when compared to controls (1.4%). Histological alterations and immature stages were observed in the testis of all exposed males; however, males exposed to EE2 were the most affected. Increased tubule number and proportionally decreased tubule diameter were observed in the testis of all EE2 groups. No effects in Vtg mRNA expression were observed in the testis; however, a significant decrease in testis ERα mRNA was observed in males exposed to 20 ng l⁻¹ TB. The groups exposed to EE2 showed a significant increase in plasma Vtg (>300-fold), hepatic Vtg mRNA (>450-fold), and ERα mRNA (>100-fold) when compared to controls. This study shows that lower concentrations of 17β-trenbolone are unable to counteract the EE2 estrogenic effects when the exposure is simultaneous.      

Efficient synthesis of an optically pure β-bromo-β,β-difluoroalanine derivative,a general precursor for β,β-difluoroamino acids

A highly efficient enantioselective preparation of a β-bromo-β,β-difluoroalanine derivative, a promising general precursor for optically active β,β-difluoroamino acids, is described. Alkylation of the hydroxypinanone glycinate Schiff base 2a with CF₂Br₂ resulted in the production of undesired dehydrobrominated product 4a promoted by the lithium alkoxide moiety on the hydroxypinanone chiral auxiliary. Dehydrobromination was prevented by protection of the hydroxyl group on the chiral auxiliary. Utilization of TMSOTf resulted in production of (S,S,S,S)-3b as the sole diastereomer without dehydrobromination. Further transformations of (S,S,S,S)-3b to other optically active β,β-difluoroamino acids were demonstrated.     

Decomposition of benzenediazonium fluoborate in esters of α,β-unsaturated acids

Summary It was shown that In contrast to the Meerwein-Koelsch reaction, the decomposition of benzenediazonlum fluoborate In esters of ,-unsaturated acids proceeds by a heterolytlc mechanism with the phenyl attacking the -carbon and the carbalkoxyl group of these esters. The reaction with ethyl crotonate gave -methylatropic acid (after hydrolysis) and a small amount of a dibasic acid, C₁₄H₁₆O₄ (or C₁₄H₁₄O₄). With methyl acrylate it gave atropic acid, phenyl acrylate, and a smaller amount of -methyl--phenylglutaric acid. With methyl methacrylate It formed benzylacrylic acid and the product of further conversions of phenyl methacrylate, namely the keto phenol 2-methyl-5-hydroxyindan-1-one.     

SPE and HPLC monitoring of 17-β-estradiol in Egyptian aquatic ecosysetms

Solid-phase extraction and HPLC methods are described for monitoring of 17-β-estradiol residues in Egyptian aquatic ecosystems (water, fish, mollusks, sediment, and drinking water) at the Nile River, Suez Canal region, and northeast of Egypt. Molecular imprinted polymer was prepared and used in extraction. High performance liquid chromatography (HPLC) columns used were Supelcosil C₁₈ and Nucleosil C₁₈. The mobile phases used were different combinations of water and acetonitrile. The concentration of 17-β-estradiol in water, aquatic animals, and sediment samples were of 265.13–7988.12 µg/L, 0.503–96.167, and 0.775–11.884 µg/kg, respectively. Marine lake was contained with high levels of 17-β-estradiol (P < 0.05). Similarly, the Nile River downstream showed high levels of 17-β-estradiol. The detected concentrations in mollusks were significantly higher than those detected in fish. Tilapia fish did not show 17-β-estradiol. Contrarily, low concentrations were detected in the rivulet streams supplied by the Nile River. Besides, 17-β-estradiol was also detected in the sediments at low levels. Detection of 17-β-estradiol in the Egyptian ecosystems attracted attention toward heavy reliance on some esterogenic medicinal products in Egypt. The monitoring of 17-β-estradiol in other water bodies was recommended. Besides, the development of methodologies of bioremediation to eliminate 17-β-estradiol from the Egyptian and other water resources of the world was also suggested.     

An unexpected synthesis of an α,β,γ,δ-unsaturated ketone due to an abnormal opening of benzylidene acetal by bromide anion

An unexpected synthesis of an α, β, γ,δ-unsaturated ketone, which embodies a new type of oxy-carbon cyclic structure, was achieved, while we tried to construct the taxane CB-ring system. Also, a series of abnormal reaction phenomena was found to be related to the formation and reaction of seven membered cyclic benzylidene acetal.     

Lignin-type, α,β-unsaturated aldehydes of lignin-type in organic solvents

A 1:1 reaction of [HO(CH₂)₃]₃P with 4-hydroxy-3-methoxy-cinnamaldehyde (coniferaldehyde) or 3,5-dimethoxy-4-hydroxycinnamaldehyde (sinapaldehyde) in acetone at room temperature affords phosphonium zwitterions of the type R₃P⁺CH(4-O^?-Ar)CH₂CHO; other phosphines [R = Et, n-Bu, (CH₂)₂CN, and p-Tol] do not react under the same conditions. In alcohols R??OH(D) [R?? = CD₃, Et, (CD₃)₂CD, s-Bu, HOCH₂CH₂], the above phosphines (except the cyano-derivative) and those where R = i-Pr, Cy, Me₂Ph, MePh₂ do react within an equilibrium established between the reactants and the zwitterion-hemiacetal products R₃P⁺CH(4-O^?-Ar)CH₂CH(OH)(OR??) that are formed as a mixture of two diastereomers. The nature of the phosphine and the alcohol affects the equilibrium and the diastereomeric ratio.     

Complete chromatographic separation of steroids, including 17α and 17β-estradiols, using a carbazole-based polymeric organic phase in both reversed and normal-phase HPLC

Poly(2-N-carbazolylethyl acrylate) with terminal trimethoxysilyl groups was prepared as an organic phase and immobilized onto silica. The retention behavior of the column packed with this carbazole-based polymer-immobilized silica (Sil-CEA) was investigated by using various estrogenic steroids and corticoids in both reversed-phase and normal-phase liquid chromatography. As a result, complete separation was confirmed for eight kinds of steroids with Sil-CEA. The most specific separation with Sil-CEA can be emphasized by the high separation factor (e.g., α = 1.39 in methanol–water (7:3, v/v) at 35 °C) for 17α and 17β-estradiols, one of the most difficult pairs of isomers in chromatographic separation, whereas for two kinds of commercially available polymeric ODS columns as references α = 1.01, only, under the same conditions. Because the excellent separation and retention order with Sil-CEA was maintained even in a normal-phase mobile phase such as a hexane–2-propanol, it is estimated that the CEA phase has multiple interaction mechanisms through stronger interactions such as dipole–dipole, carbonyl–π, and hydrogen bonding interactions than the hydrophobic effect expected with ODS.     

Focused ultrasound-assisted acceleration of enzymatic hydrolysis of alkylphenols and 17β-oestradiol glucuronide in fish bile

According to the European Water Framework Directive (WFD), alkylphenols, such as octylphenols and nonylphenols, and 17β-oestradiol are considered as priority or emerging pollutants, respectively, mainly due to their possible properties as endocrine-disrupting compounds (EDCs). EDCs are accumulated in liver, fat, kidney and bile in the glucuronide form. In order to determine the concentration of these compounds in bile, an enzymatic hydrolysis step is necessary. This step is usually long (～16 h), and in this sense, ultrasound probes were studied as a possible alternative energy source to accelerate this process. Enzymatic hydrolysis was reduced to 20 min using an ultrasound probe at one cycle and 10% of amplitude. For validation of analytical procedure, nonylphenol glucuronide (4NP-G), 4-tert-octylphenol glucuronide (4tOP-G) and 4-n-octylphenol glucuronide (4nOP-G) were synthesised while 17β-oestradiol glucuronide (E2-G) was commercially available. Bile from thick-lip grey mullets (Chelon labrosus) was spiked with known amounts of 4NP-G, 4tOP-G, 4nOP-G and E2-G and submitted to the optimised procedure. Good recoveries (77–122%), precision in the 5% to 12% range and limits of detection, ranging from the low nanogramme per gramme level for 4tOP, 4nOP and E2 to the low microgramme per gramme level for nonylphenols, were obtained. The optimised method was applied for the determination of alkylphenol in the bile of thick-lip grey mullets fish bile from the Urdaibai estuary (UNESCO reserve of the Biosphere, Bay of Biscay), and high concentrations (2.3–14.2 μg/g), such as those obtained in polluted areas, were measured. E2 was determined in the bile of thick-lip grey mullets, intraperitoneally injected with E2.     

Ferrocenyl-substituted α,β-unsaturated ketones in synthesis of tetrahydropyrimidinones

Ferrocenyltetrahydropyrimidin-2-ones were prepared by reactions of linear and cyclic ,-unsaturated ketones of the ferrocene series with urea in i-PrOH in the presence of t-BuOK. The structures of the compounds prepared were studied by ¹H and ¹³C NMR and IR spectroscopy, and also by single crystal X-ray diffraction.Translated from Zhurnal Obshchei Khimii, Vol. 74, No. 11, 2004, pp. 1887–1893.Original Russian Text Copyright © 2004 by E. Klimova, Lorez, T. Klimova, Garcia, Hernandez, Ramirez.This revised version was published online in April 2005 with a corrected cover date.     

Three steroids with unique structural feature of 5β-Spirostan-1β,3β,17α-trihydroxyl from Reineckia carnea

A new spirostanol sapogenin and two spirostanol saponins, tentatively named reineckiagenin A (1), reineckiagenoside A (2), and reineckiagenoside B (3), were isolated from the whole plant of Reineckia carnea. By detailed analysis of their 1D and 2D NMR spectra, chemical methods, and by comparison with spectra data of known compounds, the structures of the new steroids were determined to be 25(S)-5β-spirostan-1β,3β,17α-triol (1), 25(S)-5β-spirostan-1β,3β,17α-triol 1-O-β-D-xylopyranoside (2), 25(S)-5β-spirostan-1β,3β,17α-triol 1-O-α-L-rhamnopyranosyl-(1→2)-β-D-xylopyranoside (3). Compounds 1, 2, and 3 are the first naturally occurring steroids with unique structural feature of 5β-spirostan-1β,3β,17α-trihydroxyl.