Analytical Bioscience
The Centre for Analytical Biosciences is part of the School of Pharmacy at the University of Nottingham, which the 2008 Research Assessment Exercise confirmed as the UK's premier school of pharmacy, with 95% of our research judged to be of international quality and 35% world-leading. This result recognises the outstanding quality of the innovative and exciting research that is carried out in the School by a dedicated team of people.
We apply advanced analytical approaches to investigate complex biological and/or pharmaceutical problems. The majority of our studies involve the need to analyse multi-component mixtures of molecules in human, mammalian, bacterial, plant or food samples. Mass spectrometry (MS) coupled with HPLC, GC and capillary electrophoresis and NMR spectroscopy are the key techniques used. (Click here for laboratory infrastructure details) A major area of investigation is the role of biological metabolites in important biological processes and disease states, particularly using metabolomics. Our research is directly integrated with interdisciplinary projects across the university or by way of collaborative ventures with other institutions and industry.
CAB Group Photo
Exploring and exploiting advanced methods for bioanalysis
Global and targeted metabolite profiling (metabolomics) technologies
Dietary and lifestyle influences on health and disease
Pharmaceutical and therapeutic applications of metabolomics
Metabolomics applied to bacterial metabolism and quorum sensing
Equipment and facilities
Exploring and exploiting advanced methods for bioanalysis (DA Barrett, CA Daykin)
We have pioneered the development of a novel NMR-based metabolomic approach, which we have termed “ Interactive Metabolomics ” or i-metabolomics. This methodology allows the study of metabolite interactions with macromolecules in complex heterogeneous biofluids such as blood plasma. Further information and downloadable resources resulting from this EPSRC funded work are available by following the link.
We have pioneered the application of LC coupled with quadrupole linear ion trap (QTRAP) MS to provide a novel way to profile structurally related families of metabolites. Nanoflow LC-MS/MS and capillary electrophoresis-laser induced fluorescence (CE-LIF) give us exceptional facilities for high-sensitivity measurement of metabolites, eg in amino acid analysis of nanolitre volumes of phloem sap from Arabidopsis thaliana (University of Birmingham, Biosciences ).
MS-based methods to analyse the surfaces of pharmaceutical and biological samples under ambient conditions have been developed, including the novel PADI-MS (below left, developed in collaboration with Chemistry and Hiden Analytical ), DESI-MS (below right) and 'matrixless' MALDI-Tof MS (SALDI, DIOS).
Ratcliffe LV et al. Anal. Chem. (In Press) . Zhu, X et al. (2005). Electrophoresis, 26, 911 . Hunt, EJ et al. (2006) Molecular Ecology 15: 4203 .
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Global and targeted metabolite profiling (metabolomics) technologies (DA Barrett, CA Daykin)
We use NMR spectroscopy and MS based global and targeted methodologies to provide a comprehensive coverage of the metabolome. To ensure the reliability of the data required for performing metabolomics analysis we validate all stages of our methods including sample preparation and quantification. Group members are also involved in the international efforts from the metabolomics community to define reporting standards. Detailed methods for global profiling by NMR and LC-QTof-MS of blood plasma, serum, urine, sweat, liver, saliva and brain are available. Targeted profiling methods for a range of metabolites are in use including folate species, fatty acids, eicosanoids (EETS, HETEs and di-HETEs), SAM cycle metabolites, and others.
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Typical bacterial nucleotide profile produced by mass spectrometry
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Typical 800 MHz
NMR spectrum of blood plasma.
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C.A. Daykin and F. Wulfert (2006), Frontiers in Drug Design and Discovery, Vol 2.
J.C. Lindon et al. Nature Biotechnology, 23: 833.
J.L. Griffin et al. (2007) Metabolomics, In press.
L.W. Sumner et al (2007), Metabolomics, In press.
Garratt, LC (2005) Rapid Comm. Mass Spectrom, 19, 239
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Examples of applications of these methods are given below.
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Dietary and lifestyle influences on health and disease (CA Daykin, DA Barrett)
NMR spectroscopy-based metabolomics is applied to the study of nutritional intervention on human health and the composition of food products (with Unilever and School of Biosciences ). The table below shows a study of the metabolic effects of black tea, green tea and caffeine on healthy volunteers. In vitro NMR spectroscopy is applied to understanding the biochemical basis for disease: biomarker discovery for polycystic ovary syndrome with the School of Human Development ; the combination of MRI, behavioural studies and NMR-based metabolomics to accurately diagnose early stage Alzheimer�s disease (with Institute of Neuroscience ) and the relationship between poor early growth and the risk of type 2 diabetes in later life with Addenbrooks Hospital . Application of high sensitivity NMR and MS-based methods in collaboration with Unilever , aims to provide novel insights into the composition of apocrine sweat secretions, thus leading to new biological insights into the role of bacterial metabolism in underarm malodour generation.
C.A. Daykin et al (2005) J. Agric. Food Chem., 53: 1428. F.A. van Dorsten et al. (2006) J. Agric. Food Chem., 54: 6929. C.A. Daykin, et al (2007) Magnetic Resonance in Food Science: From Molecules to Man, p 36. Harker, H. et al (2006), Metabolomics, 2: 105.
LC-MS/MS-based metabolite pathway profiling methods underpin molecular level investigations of the anti-cancer mechanism of broccoli isothiocyanates (Institute of Food Research ) and folate biosynthetic pathways in plants (Biosciences ).
Al Janobi, AA et al. (2006) J. Chromatogr. B 844: 223 . Garratt, LC et al. (2005) Rapid Comm. Mass Spectrom, 19, 2390 .
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Pharmaceutical and therapeutic applications of metabolomics (DA Barrett)
LC-MS/MS and LC-QTRAP-MS methodologies are in current to profile endogenous cannabinoids and their metabolites in tissue and brain regions. The mechanisms underlying the analgesic effects of endocannabinoids in acute and chronic pain states, including neuropathic pain and osteoarthritis are being investigated with a view to finding new targets for pharmacological evaluation (with Institute of Neuroscience ). Profiling of lipid mediators and their metabolites in human uterine tissue is being used to assess whether these lipids have a role in triggering premature births (with Human Development ).
Jhaveri MD et al. (2006). J. Neuroscience 26: 13318 . Richardson D et al. (2007) Analytical Biochemistry 360: 216 .
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Metabolomics applied to bacterial metabolism and quorum sensing (DA Barrett)
Metabolite profiling methods specially designed for comprehensive screening of families of quinolone, homoserine lactone and autoinducer quorum sensing (QS) molecules, are being used to discover new QS molecules and to investigate the relationship between bacterial central metabolism and quorum sensing (illustrated in schematic to left) using bioreactor/chemostat bacterial metabolomics methodology (one of several collaborartions with the Institute of Infections and Immunity ). Image to right shows a LC-QTRAP-MS method for simultaneous profiling of three QS molecule �families� in Pseudomonas aeruginosa .
Diggle P et al. (2006) Chemistry & Biology 13: 701 .
Ortori, CA et al. (2007) Analytical and Bioanalytical Chem. 387: 497 .
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Funded by: BBSRC, EPSRC, MRC, Wellcome Trust, Action Medical Research and Industry
Academic staff
DA Barrett - Associate Professor and Reader in Analytical Bioscience
CA Daykin - Lecturer in Analytical Bioscience
Senior Experimental Officer
Dr CA Ortori