Steroidal estrogens in Phaseolus vulgaris

Young, Ian James (1977) Steroidal estrogens in Phaseolus vulgaris. PhD thesis, University of Glasgow.

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Abstract

This thesis presents an investigation into the presence of steroidal estrogens in Phaseolus vulgaris var. Canadian Wonder. The approach was threefold. Firstly purification methods were appraised, and it was found that partitioning ether with 1N NaOH removed ca 60% of [14C]-estradiol compared with only ca 40% with 2N and 30% with 4N NaOH. Partitioning 1N NaOH against a benzene : butanol mixture was also only ca 40% efficient. Suitable preparative chromatographic systems - thin-layer chromatography and elution on a hydroxyalkoxypropyl derivative of Sephadex - were developed. Secondly, employing the most efficient of these methods, evidence of estradiol in extracts of seeds, vegetative and flowering plants using a modified radioimmunoassay technique and combined gas chromatography- mass spectrometry was obtained. The GC-MS analyses involved one of two methods: either continuous monitoring at the collector of the ion abundance at m/e 416 (the molecular ion of the bis-trimethylsilyl derivative of estradiol) as generated in the ion source from the GLC effluent of two different columns, allowing the mass spectrometer to be used as a semispecific detector for estradiol; or three ions (m/e 416, 342, 285) were continuously monitored in a process commonly referred to as mass fragmentography - a sensitive and selective detector system. (M/e 342 is the molecular ion of estrone trimethylsilyl ether, and m/e 285 is the base ion common to both E1 and E2). In the third approach an investigation was carried out to ascertain whether the dwarf French bean could synthesise estrogens. Eight-day old seedlings were sectioned at the hypocotyl, under water, and stood in aqueous solutions of various radioactive substances - [14C]-estrone, estradiol and mevalonic acid as well as [3H]-estrone sulphate. Chromatographic mobilities of metabolites were compared with authentic compounds using TLC on three solvent systems, column chromatography and radio-gas-liquid chromatography of the trimethylsilyl derivative. Samples believed to contain estradiol were diluted with non-radioactive carrier estradiol and recrystallised to constant specific activity. Using these techniques in combination, radioactivity was incorporated into estradiol from estrone, estrone sulphate and mevalonic acid. There is also some evidence from thin-layer and column chromatography that estradiol is converted to estrone.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Additional Information: Adviser: J R Hillmen
Keywords: Plant sciences
Date of Award: 1977
Depositing User: Enlighten Team
Unique ID: glathesis:1977-73131
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 14 Jun 2019 08:56
Last Modified: 14 Jun 2019 08:56
URI: http://theses.gla.ac.uk/id/eprint/73131

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