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The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP. / Shujing Ding

Swansea University Author: Shujing Ding

Abstract

Mass spectrometry is widely used nowadays especially in the fields of pharmaceutical and proteomics research. Ginkgo biloba is one of the top selling phytophamiaceuticals in the US and Europe. The two major active components of Ginkgo leaf extract are the flavonoids and terpene lactones. Identificat...

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Published: 2006
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
URI: https://cronfa.swan.ac.uk/Record/cronfa42438
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Identification, determination, as well as the physiological effects of these two sets of compounds have been of increasing interest over the last 20 years. In this thesis, systematic qualitative and quantitative studies of the flavonoids and terpene lactones in Ginkgo biloba by liquid chromatography / mass spectrometry have been undertaken. Also in this thesis, mass spectrometric methodology was developed and applied to the identification of the proteins specifically phosphorylated in response to cCMP. Structural information of Ginkgo biloba flavonoids and terpene lactones, the fragment of compounds were obtained on both a LCQ ion trap and Q-TOF mass spectrometer. The tentative fragment pathways were proposed and used for structural elucidation of some unknown components in Ginkgo biloba commercial products. Capillary column separation of Ginkgo biloba commercial product was evaluated and fingerprint profiles of five Ginkgo biloba commercial products were compared. A reverse-phase high-performance liquid chromatography electrospray ionisation (RP-HPLC/ESI) mass spectrometry method was developed and validated for the simultaneous determination of ten major active components in Ginkgo biloba extract (bilobalide, ginkgolides A, B, C, quercetin, kaempferol, isorhamnetin, rutin hydrate, quercetin-3-beta-D-glucoside and quercitrin hydrate). The quantitative determination of flavonoids and terpene lactones by LC/MS in human urine after consumption of Ginkgo biloba product was developed. The online solid-phase extraction and capillary column with column-switch technique require minimum sample pre-treatment and both flavonoids and terpene lactones can be detected simultaneously. The mass accuracy at high molecular weight by matrix-assisted laser desoiption/ionisation time-of-flight mass spectrometry was investigated to resolve a question on mass accuracy which had been observed to be relatively low for high mss proteins. Bovine serum albumin (BSA) was employed as a model compound and strategies to improve mass measurement at high mass were examined. LC/MS was applied in part of the cyclic nucleotide project in the School of Biological Science. Since cAMP and cGMP are recognized second messengers and play important roles in signal transduction, to elucidate the function of cCMP in signal transduction, efforts were made to identify the cCMP-responsive protein kinase substrates. 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spelling 2018-08-02T16:24:29.2597915 v2 42438 2018-08-02 The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP. dd0c2a491755c50e20a003e1005a5e85 NULL Shujing Ding Shujing Ding true true 2018-08-02 Mass spectrometry is widely used nowadays especially in the fields of pharmaceutical and proteomics research. Ginkgo biloba is one of the top selling phytophamiaceuticals in the US and Europe. The two major active components of Ginkgo leaf extract are the flavonoids and terpene lactones. Identification, determination, as well as the physiological effects of these two sets of compounds have been of increasing interest over the last 20 years. In this thesis, systematic qualitative and quantitative studies of the flavonoids and terpene lactones in Ginkgo biloba by liquid chromatography / mass spectrometry have been undertaken. Also in this thesis, mass spectrometric methodology was developed and applied to the identification of the proteins specifically phosphorylated in response to cCMP. Structural information of Ginkgo biloba flavonoids and terpene lactones, the fragment of compounds were obtained on both a LCQ ion trap and Q-TOF mass spectrometer. The tentative fragment pathways were proposed and used for structural elucidation of some unknown components in Ginkgo biloba commercial products. Capillary column separation of Ginkgo biloba commercial product was evaluated and fingerprint profiles of five Ginkgo biloba commercial products were compared. A reverse-phase high-performance liquid chromatography electrospray ionisation (RP-HPLC/ESI) mass spectrometry method was developed and validated for the simultaneous determination of ten major active components in Ginkgo biloba extract (bilobalide, ginkgolides A, B, C, quercetin, kaempferol, isorhamnetin, rutin hydrate, quercetin-3-beta-D-glucoside and quercitrin hydrate). The quantitative determination of flavonoids and terpene lactones by LC/MS in human urine after consumption of Ginkgo biloba product was developed. The online solid-phase extraction and capillary column with column-switch technique require minimum sample pre-treatment and both flavonoids and terpene lactones can be detected simultaneously. The mass accuracy at high molecular weight by matrix-assisted laser desoiption/ionisation time-of-flight mass spectrometry was investigated to resolve a question on mass accuracy which had been observed to be relatively low for high mss proteins. Bovine serum albumin (BSA) was employed as a model compound and strategies to improve mass measurement at high mass were examined. LC/MS was applied in part of the cyclic nucleotide project in the School of Biological Science. Since cAMP and cGMP are recognized second messengers and play important roles in signal transduction, to elucidate the function of cCMP in signal transduction, efforts were made to identify the cCMP-responsive protein kinase substrates. Methodology of specific enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) was developed, phosphorylated proteins responding specifically to cCMP were proposed, and this supports the relationship of cCMP with cell hyperproliferation. E-Thesis Analytical chemistry. 31 12 2006 2006-12-31 COLLEGE NANME Chemistry COLLEGE CODE Swansea University Doctoral Ph.D 2018-08-02T16:24:29.2597915 2018-08-02T16:24:29.2597915 Faculty of Science and Engineering School of Engineering and Applied Sciences - Chemistry Shujing Ding NULL 1 0042438-02082018162454.pdf 10798146.pdf 2018-08-02T16:24:54.4230000 Output 12739680 application/pdf E-Thesis true 2018-08-02T16:24:54.4230000 false
title The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
spellingShingle The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
Shujing Ding
title_short The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
title_full The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
title_fullStr The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
title_full_unstemmed The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
title_sort The application of mass spectrometry to Ginkgo biloba analysis and identification of phosphorylated proteins in response to elevated level of cCMP.
author_id_str_mv dd0c2a491755c50e20a003e1005a5e85
author_id_fullname_str_mv dd0c2a491755c50e20a003e1005a5e85_***_Shujing Ding
author Shujing Ding
author2 Shujing Ding
format E-Thesis
publishDate 2006
institution Swansea University
college_str Faculty of Science and Engineering
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hierarchy_top_id facultyofscienceandengineering
hierarchy_top_title Faculty of Science and Engineering
hierarchy_parent_id facultyofscienceandengineering
hierarchy_parent_title Faculty of Science and Engineering
department_str School of Engineering and Applied Sciences - Chemistry{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Engineering and Applied Sciences - Chemistry
document_store_str 1
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description Mass spectrometry is widely used nowadays especially in the fields of pharmaceutical and proteomics research. Ginkgo biloba is one of the top selling phytophamiaceuticals in the US and Europe. The two major active components of Ginkgo leaf extract are the flavonoids and terpene lactones. Identification, determination, as well as the physiological effects of these two sets of compounds have been of increasing interest over the last 20 years. In this thesis, systematic qualitative and quantitative studies of the flavonoids and terpene lactones in Ginkgo biloba by liquid chromatography / mass spectrometry have been undertaken. Also in this thesis, mass spectrometric methodology was developed and applied to the identification of the proteins specifically phosphorylated in response to cCMP. Structural information of Ginkgo biloba flavonoids and terpene lactones, the fragment of compounds were obtained on both a LCQ ion trap and Q-TOF mass spectrometer. The tentative fragment pathways were proposed and used for structural elucidation of some unknown components in Ginkgo biloba commercial products. Capillary column separation of Ginkgo biloba commercial product was evaluated and fingerprint profiles of five Ginkgo biloba commercial products were compared. A reverse-phase high-performance liquid chromatography electrospray ionisation (RP-HPLC/ESI) mass spectrometry method was developed and validated for the simultaneous determination of ten major active components in Ginkgo biloba extract (bilobalide, ginkgolides A, B, C, quercetin, kaempferol, isorhamnetin, rutin hydrate, quercetin-3-beta-D-glucoside and quercitrin hydrate). The quantitative determination of flavonoids and terpene lactones by LC/MS in human urine after consumption of Ginkgo biloba product was developed. The online solid-phase extraction and capillary column with column-switch technique require minimum sample pre-treatment and both flavonoids and terpene lactones can be detected simultaneously. The mass accuracy at high molecular weight by matrix-assisted laser desoiption/ionisation time-of-flight mass spectrometry was investigated to resolve a question on mass accuracy which had been observed to be relatively low for high mss proteins. Bovine serum albumin (BSA) was employed as a model compound and strategies to improve mass measurement at high mass were examined. LC/MS was applied in part of the cyclic nucleotide project in the School of Biological Science. Since cAMP and cGMP are recognized second messengers and play important roles in signal transduction, to elucidate the function of cCMP in signal transduction, efforts were made to identify the cCMP-responsive protein kinase substrates. Methodology of specific enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) was developed, phosphorylated proteins responding specifically to cCMP were proposed, and this supports the relationship of cCMP with cell hyperproliferation.
published_date 2006-12-31T03:52:58Z
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