Date of Award

Summer 1990

Document Type


Degree Name

Master of Science (MS)


Chemistry & Biochemistry



Committee Director

Patricia A. Pleban

Committee Member

Frank E. Scully, Jr.

Committee Member

Anibal Acosta

Call Number for Print

Special Collections LD4331.C45K47


Triphenylethylene antiestrogen compounds are used to stimulate ovulation in anovulary women and spermatogenesis in oligozoospermatic males. Clomiphene Citrate (CC) is given therapeuticaly as a mixture of the trans (E) and cis (Z) isomers. Each of these isomers has its own characteristic agonist-antagonist profiles. Only the trans form of Tamoxifen (T) is administered therapeutically.

HPLC methodologies were developed for the determination of CC and Tin human blood and seminal plasma. The methodologies involved extraction of the triphenylethylene compounds using hexane, followed by evaporation of the hexane under N2. The residue was then taken up in a volume of HPLC grade methanol and injected onto a reverse-phase octyl column, and eluted using a mobile phase consisting of methanol, H2O, H3PO4, and triethylamine.

Separation of the cc isomers was achieved; and T, as well as several of its metabolites, was assayed in human semen and serum. Following separation on the column, the eluted compounds were transformed into fluorescent stilbene compounds using post column UV irradiation and detected using an on-line fluorescence detector. Investigations of the effects of pH, the triethylamine concentration, the total phosphate concentration, and polarity of the mobile phase on the separation of T and the isomers of cc were done. In addition, the effects of the exposure of specimen to light on the lability of T were determined.

The methodologies gave a precision range of 6 to 19 % CV and a detection limit of 1.3 ng, making them suitable for assay of triphenylethylene compounds in semen and blood serum.


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