Resolution of equine estrogens using glass capillary columns

Pillai, Gopal S; McErlane, Keith M.

doi:10.1002/jhrc.1240040205

J. High Resol. Chromatogr.

1981

DOI: 10.1002/jhrc.1240040205

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Resolution of equine estrogens using glass capillary columns

Gopal S Pillai

Keith M. McErlane

Abstract: SummaryA rapid and specific procedure is described for the resolution of nine steroids present in conjugated estrogen preparations. Resolution is achieved on a short glass capillary column coated with Silar 1 OC after dual derivatization to oxime-trimethylsilyl ethers.

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Cited by 8 publications

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References 22 publications

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“…Using a 15 meter Silar 1OC glass capillary column all nine components were adequately separated as their oxime-trimethylsilyl derivatives within 30 minutes [14]. Packed column gas chromatography-chemical ionization mass spectrometry of the trifluoroacetyl derivatives hasalso been used to quantify all of the constituent estrogens present in seven pharmaceutical preparations [K].…”

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confidence: 99%

Measurement of equilin and estrone in human plasma by capillary gas chromatography‐negative ion chemical ionization mass spectrometry

Robinson¹,

Jones²,

Maddock³

1987

J. High Resol. Chromatogr.

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Capillary gas chromatography Negative ion mass spectrometry Plasma Estrone, equilin Conjugated estrogens A procedure is described for the analysis of the estrogens equilin and estrone in human plasma following oral administration of conjugated estrogen preparations. After enzymatic hydrolysis of the sulfate conjugates, plasma proteins are precipitated with methanol and the estrogens extracted into ethyl acetate. Derivatization with the reagent flophemesylamine converts equilin and estrone into volatile pentafluorophenyldimethylsilyl ethers ideally suited to capillary gas chromatography-negative ion chemical ionization mass spectrometry. Using a 15 meter dimethyl silicone bonded phase fused silica capillary column separation of the estrone and equilin derivatives is achieved within 9 minutes. Selected ion monitoring of the intense negative molecular ions enables levels of 1 ng.ml-' to be measured with coefficients of variation of 9.3 %and 14.2 % for estrone and equilin respectively. Plasma levels of the compounds are reported in two male volunteers up to 24 hours after dosing with 5 milligrams of PremarinTM. (TM Ayerst Laboratories Inc., New York, USA.) Conjugated estrogen preparations have been in use for many years as replacement therapy for the treatment of symptoms associated with menopause [l]. The pharmaceutical product is formulated from extracts of pregnant mares' urine and is a mixture of the sulfate esters of the closely related compounds estrone, equilin, equilenin, 17 a-estradiol, 17 B-estradiol, 17 a-dihydroequilin, 17 0-dihydroequilin, 17 a-dihydroequilenin, and 17 P-dihydroequilenin [2,3].The relative amounts of each compound present in the formulation can vary according to the time of collection of the mares' urine. However, the major constituents as defined by the United States Pharmacopeia [4] are sodium estrone sulfate (50% -60%) and sodium equilin sulfate (22.5% -32.5%). Other preparations known as esterified estrogens contain different proportions of the individual estrogens but still with the estrone and equilin esters predominant [4,5].

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mentioning

confidence: 99%

Measurement of equilin and estrone in human plasma by capillary gas chromatography‐negative ion chemical ionization mass spectrometry

Robinson¹,

Jones²,

Maddock³

1987

J. High Resol. Chromatogr.

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Quantitative determination of conjugated estrogens in formulations by capillary GLC

Pillai

McErlane

1981

Journal of Pharmaceutical Sciences

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Separation of steroid isomers using a liquid crystalline polysiloxane stationary phase in capillary supercritical fluid chromatography

Chang

Markides

Bradshaw

et al. 1989

J Microcolumn Separations

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Abstract.A liquid crystalline biphenylcarboxylate ester polysiloxane stationary phase was used to study the effect of solute structure on retention for the separation of isomeric steroids in capillary supercritical fluid chromatography. It was determined that the retention mechanism is related to solute molecular shape rather than to properties such as volatility or polarity. The elution order of isomeric androstanes and estrogens could be correlated with the planarity of molecules: more-planar molecules were retained longer on the column. The resolution of steroid isomers was improved significantly using the liquid crystalline stationary phase when compared with results obtained using other stationary phases.

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Resolution of equine estrogens using glass capillary columns

Abstract: SummaryA rapid and specific procedure is described for the resolution of nine steroids present in conjugated estrogen preparations. Resolution is achieved on a short glass capillary column coated with Silar 1 OC after dual derivatization to oxime-trimethylsilyl ethers.

Cited by 8 publications

References 22 publications

Measurement of equilin and estrone in human plasma by capillary gas chromatography‐negative ion chemical ionization mass spectrometry

Measurement of equilin and estrone in human plasma by capillary gas chromatography‐negative ion chemical ionization mass spectrometry

Quantitative determination of conjugated estrogens in formulations by capillary GLC

Separation of steroid isomers using a liquid crystalline polysiloxane stationary phase in capillary supercritical fluid chromatography

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