Analysis of raspberry ketone and related compounds from plant cell suspension cultures
This thesis is constructed as a part of a larger research project aiming to increase understanding of polyketone reductases (PKR) and develop applications from them. PKRs are enzymes in biosynthetic pathways leading to several aromatic secondary metabolites in plants. The previous work in the resear...
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Helsingfors universitet
2012
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ftunivhelsihelda:oai:helda.helsinki.fi:10138/36666 2023-08-20T04:05:02+02:00 Analysis of raspberry ketone and related compounds from plant cell suspension cultures Selin, Markus Helsingin yliopisto, Farmasian tiedekunta University of Helsinki, Faculty of Pharmacy Helsingfors universitet, Farmaceutiska fakulteten 2012 http://hdl.handle.net/10138/36666 eng eng Helsingfors universitet University of Helsinki Helsingin yliopisto URN:NBN:fi-fe201801151192 http://hdl.handle.net/10138/36666 PKR BAR Rubus extraction chemical analysis raspberry ketone zingerone vatukat uutto kemiallinen analyysi vadelman ketoni tsingeroni Farmakognosi Pharmacognosy Farmakognosia pro gradu -avhandlingar pro gradu -tutkielmat master's thesis 2012 ftunivhelsihelda 2023-07-28T06:10:12Z This thesis is constructed as a part of a larger research project aiming to increase understanding of polyketone reductases (PKR) and develop applications from them. PKRs are enzymes in biosynthetic pathways leading to several aromatic secondary metabolites in plants. The previous work in the research group has led to establishment of several callus cultures from plants belonging to the genus Rubus in the family Rosaceae. The aim in the experimental part of this thesis is the identification and semi-quantitation of raspberry ketone (RK) and related aromatics in the cell suspension cultures initiated from the previously established callus cultures. RK is biosynthetically produced by reduction of p-hydroxybenzalacetone (p-OH-BA) by benzalacetone reductase (BAR). As a part of the experimental work, p-OH-BA has to be chemically synthetized and analysed. Special emphasis is placed to experiment, develop and validate an extraction method for phenolic compounds using ASE 200 working station. In the review part of this thesis, the basic procedures of chemical analysis are described, optimization and validation of analytical methods are discussed, and lastly studies related to raspberry ketone (RK) are summarized. The detection limit is 0.73 µg/ml for RK with the established UPLC-UV method, and the quantitation limit (QL) is 2.22 µg/ml. At the QL, the standard deviation of the extraction method is 8.9 % and the results are 6.4 % higher than expected. At the high end of the standard curve the extraction results are 18.7 % higher than expected. Some changes are proposed to optimize the method. Analysis of the cell line extracts with the established UPLC-UV method did not readily reveal any of the studied compounds. Although the interpretation of the results of the MS experiment is still underway, RK was detected from the arctic bramble cell line Ra15. Also, a possible derivative of zingerone was detected from cloudberry cell line extract even without the corresponding standard compound. This shows the power of the MS in ... Master Thesis Arctic Helsingfors Universitet: HELDA – Helsingin yliopiston digitaalinen arkisto Arctic |
institution |
Open Polar |
collection |
Helsingfors Universitet: HELDA – Helsingin yliopiston digitaalinen arkisto |
op_collection_id |
ftunivhelsihelda |
language |
English |
topic |
PKR BAR Rubus extraction chemical analysis raspberry ketone zingerone vatukat uutto kemiallinen analyysi vadelman ketoni tsingeroni Farmakognosi Pharmacognosy Farmakognosia |
spellingShingle |
PKR BAR Rubus extraction chemical analysis raspberry ketone zingerone vatukat uutto kemiallinen analyysi vadelman ketoni tsingeroni Farmakognosi Pharmacognosy Farmakognosia Selin, Markus Analysis of raspberry ketone and related compounds from plant cell suspension cultures |
topic_facet |
PKR BAR Rubus extraction chemical analysis raspberry ketone zingerone vatukat uutto kemiallinen analyysi vadelman ketoni tsingeroni Farmakognosi Pharmacognosy Farmakognosia |
description |
This thesis is constructed as a part of a larger research project aiming to increase understanding of polyketone reductases (PKR) and develop applications from them. PKRs are enzymes in biosynthetic pathways leading to several aromatic secondary metabolites in plants. The previous work in the research group has led to establishment of several callus cultures from plants belonging to the genus Rubus in the family Rosaceae. The aim in the experimental part of this thesis is the identification and semi-quantitation of raspberry ketone (RK) and related aromatics in the cell suspension cultures initiated from the previously established callus cultures. RK is biosynthetically produced by reduction of p-hydroxybenzalacetone (p-OH-BA) by benzalacetone reductase (BAR). As a part of the experimental work, p-OH-BA has to be chemically synthetized and analysed. Special emphasis is placed to experiment, develop and validate an extraction method for phenolic compounds using ASE 200 working station. In the review part of this thesis, the basic procedures of chemical analysis are described, optimization and validation of analytical methods are discussed, and lastly studies related to raspberry ketone (RK) are summarized. The detection limit is 0.73 µg/ml for RK with the established UPLC-UV method, and the quantitation limit (QL) is 2.22 µg/ml. At the QL, the standard deviation of the extraction method is 8.9 % and the results are 6.4 % higher than expected. At the high end of the standard curve the extraction results are 18.7 % higher than expected. Some changes are proposed to optimize the method. Analysis of the cell line extracts with the established UPLC-UV method did not readily reveal any of the studied compounds. Although the interpretation of the results of the MS experiment is still underway, RK was detected from the arctic bramble cell line Ra15. Also, a possible derivative of zingerone was detected from cloudberry cell line extract even without the corresponding standard compound. This shows the power of the MS in ... |
author2 |
Helsingin yliopisto, Farmasian tiedekunta University of Helsinki, Faculty of Pharmacy Helsingfors universitet, Farmaceutiska fakulteten |
format |
Master Thesis |
author |
Selin, Markus |
author_facet |
Selin, Markus |
author_sort |
Selin, Markus |
title |
Analysis of raspberry ketone and related compounds from plant cell suspension cultures |
title_short |
Analysis of raspberry ketone and related compounds from plant cell suspension cultures |
title_full |
Analysis of raspberry ketone and related compounds from plant cell suspension cultures |
title_fullStr |
Analysis of raspberry ketone and related compounds from plant cell suspension cultures |
title_full_unstemmed |
Analysis of raspberry ketone and related compounds from plant cell suspension cultures |
title_sort |
analysis of raspberry ketone and related compounds from plant cell suspension cultures |
publisher |
Helsingfors universitet |
publishDate |
2012 |
url |
http://hdl.handle.net/10138/36666 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_relation |
URN:NBN:fi-fe201801151192 http://hdl.handle.net/10138/36666 |
_version_ |
1774715456053051392 |