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Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method
Dissertation (MSc)--University of Pretoria, 2016.
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2016
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| _version_ | 1867613668631379968 |
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| access_status_str | Open Access |
| author2 | Cromarty, Allan Duncan |
| author_browse | Cromarty, Allan Duncan |
| author_facet | Cromarty, Allan Duncan |
| collection | Thesis |
| dc_rights_str_mv | © 2016 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc)--University of Pretoria, 2016. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/56953 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:39:48.717Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2016 |
| publishDateRange | 2016 |
| publishDateSort | 2016 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/56953 Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method Cromarty, Allan Duncan u29038172@tuks.co.za Cardeano, Joao Carlos UCTD Health sciences theses SDG-03 Health sciences theses SDG-17 Dissertation (MSc)--University of Pretoria, 2016. Introduction: Biogenic amines are a group of endogenous compounds which play an important role in maintaining homeostasis. These compounds are produced from amino acid precursors and exert strong physiological effects despite being at low concentrations (low pg/ml). There are a wide variety of affects and effects controlled by biogenic amines. Different conditions can be brought on by changes in the concentrations of these biogenic amines that can alter the levels or be brought about by incorrect levels. Thus biogenic amine concentrations may prove to be valuable biomarkers if the extremely low levels can be accurately and rapidly quantitated. The aim of this study was to develop and validate a method which is able to quantitate multiple biogenic amines in a single run from low sample volumes. Methods and Materials: Quantification of the blood levels of biogenic amines was performed following extraction using derivatisation and an appropriate liquid chromatography tandem mass spectrometric method. The derivatisation was performed at mild conditions for an hour and extracted using ethyl acetate. The LC-MS/MS system used was an Agilent 1100 series HPLC and an ABSciex 4000 QTrap. The system was operated in positive ESI mode and appropriate MRM precursor-product pairs were selected. In order to validate the method a series of 8 dilutions from 0.05 2.5 ng/ml were prepared in solvent and plasma. These were then processed according to the developed method and analysed using Analyst software. Results and discussion: The validation of the biogenic amines was performed according to the ICH guidelines. The calibration curves were fitted using linear regression (1/x weighting) over the linear range of the calibration curve. Metanephrine and melatonin were validated in plasma between 0.1 2.5 ng/ml. Serotonin, histamine and normetanephrine were validated in solvent between 0.25 2.5 ng/ml. Adrenaline, dopamine, noradrenaline and 5-hydroxy-3-indoleacetic acid were validated in solvent between 5 50 ng/ml. Metanephrine exhibited the highest sensitivity and showed linearity between 0.05 2.5 ng/ml in solvent. The catecholamines and 5-HIAA were the least sensitive of the analytes and had estimated LLOQs between 0.5 2.5 ng/ml. Stability tests show the derivatised analytes to be stable at -20°C for 5 days and recovery testing shows recoveries of over 70% for metanephrine and melatonin. Conclusion: Sensitivity still remains an issue when working with biogenic amines. The extremely low physiological concentrations are further exacerbated by the low sample volumes used in this study. Despite the challenges the compounds were all validated according to ICH guidelines and an LC-MS/MS method was developed that was able to analyse 9 biogenic amines of different classes in a single run using only 250 ?l of human plasma, after multistep sample preparation whilst achieving high sensitivity. tm2016 em2026 Pharmacology MSc Unrestricted SDG-03: Good health and well-being SDG-17: Partnerships for the goals 2016-09-26T06:59:03Z 2016-09-26T06:59:03Z 2016/09/02 2016 Dissertation Cardeano, JC 2016, Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method, MSc Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/56953> S2016 http://hdl.handle.net/2263/56953 en © 2016 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | UCTD Health sciences theses SDG-03 Health sciences theses SDG-17 Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| title | Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| title_full | Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| title_fullStr | Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| title_full_unstemmed | Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| title_short | Quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| title_sort | quantification and analysis of biogenic amines using a liquid chromatography tandem mass spectrometric method |
| topic | UCTD Health sciences theses SDG-03 Health sciences theses SDG-17 |
| url | http://hdl.handle.net/2263/56953 |