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Title: The development and evaluation of an HPLC method of analysis for nicotine and its major metabolites in urine
Author: McBride, M. B.
ISNI:       0000 0001 3622 5963
Awarding Body: University of Bristol
Current Institution: University of Bristol
Date of Award: 1988
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This dissertation details the development and evaluation of an HPLC method of analysis for nicotine and the metabolites cotinine, nicotine-1'-N-oxide and 3' hydroxycotinine in urine samples. The significance of nicotine, its absorption, metabolism and excretion in man and other animals have been described in Chapter 1. Chapter 2 deals with the development of the HPLC method of analysis using both isocratic and gradient elution with W detection. A selection of packing materials/mobile phases covering different retention mechanisms was investigated. A separation of nicotine, cotinine, nicotine- 1'-N-oxide and 3' hydroxycotinine and two chromatographic standards, N' acetyl nornicotine and 2-methyl-6-(3-pyridyl)- tetrahydro-(1,2)-oxazine was achieved on a Resolve C18 5μ radially packed cartridge using gradient elution under reverse phase partition conditions. N' acetyl nornicotine was later discarded in favour of 2-methyl-6-(3-pyridyl)-tetrahydro-(1,2)-oxazine which could be used as an internal standard. The statistical analysis of the instrument response to nicotine and its metabolites in standard solutions was examined in Chapter 3- A comparison of the measurement parameters peak height and peak area was made. Within-run and between-run precision were calculated. Calibration curves were constructed with Working-Hotelling 95% confidence bands and 95% confidence bounds for 90% of future observations. The limit of detection values were also statistically calculated. Precision was found to be low for some of the components and this was reflected in unacceptably high values of the limit of detection. The clean-up of urine samples and the extraction of the components of interest were investigated in Chapter 4. Clean-up and extraction proved to be very difficult and analyses of smokers' urine samples underlined the need for an effective clean-up procedure, efficient chromatography and a sensitive and selective method of detection. It was concluded that the developed HPLC method of analysis was inadequate for quantitative analysis of nicotine and its metabolites in urine.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available