2013 Young Investigator Award Nominee: Christopher Benton

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What drove you to choose a career in bioanalysis?

During a 13-month industrial placement at Quotient Bioresearch, I worked in the Bioanalysis sector of contract research using UPLC–MS/MS to quantify small-molecule drugs in clinical and pre-clinical matrices, for pharmacokinetic, toxicokinetic and bioequivalence studies. I was fascinated with how analytical instruments, particularly LC–MS, could provide so much information in to areas such as drug metabolism, biomarkers of disease and toxicological analysis using such complex matrices. I was automatically drawn to a career in bioanaysis due to its multi-disciplinary nature, combining chemistry and physics to investigate the effects of chemicals on biological systems, for the benefit of health.

Describe the main highlights of your bioanalytical research, and its importance to the bioanalytical community both now and in the future

Throughout my PhD and work experiences I have developed and validated robust, sensitive and specific UHPLC–MS/MS assays, using various on-line and off-line sample preparation techniques, for routine biochemical diagnosis, clinical research and a contract research organisation. In today’s laboratories it is important that assays are efficient yet environmentally friendly. As a result, I havedeveloped or replaced existing HPLC methods with UHPLC and ‘superficially porous’ particle columns, improving resolution and increasingsensitivity of detection, whilst reducing assay time and solvent consumption making them perfectly suited to low-cost and high-throughput laboratories. Most recently, during my PhD, I havefocused on using UHPLC–MS/MS to investigate the pathogenesis of disorders of the heme biosynthetic pathway, together with developing assays for routine clinical diagnosis. Until recently, analytical methods for the diagnosis of the acute hepatic porphyrias were laborious, non-specific and required convoluted sample derivatisation not suited to a routine clinical laboratory. Using HILIC–MS/MS I have developed analytical methods for the direct and rapid biochemical diagnosis of the acute hepatic porphyrias, which have replaced conventional methods and are now being implemented internationally. Furthermore, the methods have sparked interest for clinical research and are being applied to targeted metabolomics to further understand the metabolic disorders.

Describe the most difficult challenge you have encountered in the laboratory and how you overcame it?

I think one of the biggest challenges that I have faced in the laboratory and continue to face, particularly when interpreting complex mass spectra, is see what the results show you and not what you want to see. For example the consecutive loss of -44 Da in MS/MS mass spectra was actually polyethylene glycol not the loss CO2 and a methylated metabolite was actually generated in vitro by storing an acidified analyte in methanol. Throughout my BSc degree in Forensic Science we were frequently reminded about the importance of impartiality in science and from my research experiences I have understood the fundamental importance of this statement. Having witnessed and proven that research groups have mischaracterized contaminants as metabolites in high impact journals, I continue to ensure that all my results are scientifically sound using chromatographic standards, accurate MS and various analytical techniques to support results, together with advice from experts in the field.

Where do you see your career in bioanalysis taking you?

I hope to continue applying analytical methods to understand and answer important biological questions, therefore, contributing to science. I have particularly enjoyed utilizing LC–MS/MS to elucidate novel biological metabolites, identifying new metabolic pathways and biomarkers of chemical intoxication. I would like to pursue this further by using targeted metabolomics to investigate drug metabolism in biological systems. I feel this area is important because it is imperative for the future of medicine that new chemical entities continue to emerge from the process of drug discovery, however, little information is known about their pharmacokinetic and toxicokinetic parameters without bioanalysis. Furthermore, not only is bioanalyis imperative for the development of novel therapeutic drugs but it is also crucial to prevent those who choose to abuse such drugs and cheat in sport. From experiencing what advantages recent developments in analytical instruments have offered for these issues, such as accurate mass spectrometry, I look forward to applying the future developments in LC–MS to such challenges.

How do you envisage the field of bioanalysis evolving in the future?

During my PhD at King’s College Hospital I have observed how LC–MS is becoming an integral part of bioanalysis in routine clinical chemistry laboratories. The improved specificity that LC–MS offers over spectrophotometry and immunoassay, dramatically improves the accuracy of patient results and as a result patient care. I feel the specificity of LC–MS combined with immunoassay, will make targeted metabolomics and proteomics much more accurate in routine and research laboratories compared with immunoassay alone. Furthermore, I think advances in instrumentation will see LC–MS become common place in automated based hospital laboratories, dramatically increasing sample throughput. I believe ion mobility-MS will play a greater role in bioanalysis for small- and large-molecule analysis, ultimately as the resolving power of ion mobility improves. The ability to remove isobaric interferences that chromatographically co-elute and separate unresolvable isomers in milliseconds is very advantageous, particularly when coupled to high-resolution MS. Naturally, I envisage that the sensitivity of analytical instruments will increase and also that sample extraction techniques for highly hydrophilic analytes such as conjugated metabolites will improve, enabling cleaner sample extracts and therefore greater sample throughput.

Please list 5 of your recent publications, and select one that best highlights your career to date in the field of bioanalysis.

Benton CM, Couchman C, Marsden JT, Rees DC, Moniz C, Lim CK. Direct and simultaneous determination of 5-aminolaevulinic acid and porphobilinogen in urine by hydrophilic interaction liquid chromatography-electrospray ionisation/tandem mass spectrometry. Biomed. Chromatogr.y 26,1033–1040 (2012).

BentonCM, Moniz C, Lim CK,Jones DJL. Travelling wave ion mobility spectrometry mass spectrometry of 5-aminolaevulinic acid, porphobilinogen and porphyrins. Rapid Comm. Mass Spectrom. 26, 480–486 (2012).

BentonCM, Lim CK, Moniz C, Jones DJL. Travelling wave ion mobility mass spectrometry of 5-aminolaevulinic acid, porphobilinogen and porphyrins.Rapid Comm. Mass Spectrom. 26(4),480–486 (2012).

BentonCM, Lim CK, Moniz C, Jones DJL. Porphyrinogen fragmentation profiles by ultra-high-performance liquid chromatography/electrospray ionisation tandem mass spectrometry. Rapid Comm. Mass Spectrom. 25, 3749–3757 (2011).

BentonCM, Lim CK, Moniz C,Jones DJL. Ultra high-performance liquid chromatography of porphyrins in clinical materials: column and mobile phase selection and optimisation. Biomedical Chromatography 26, 714–719 (2011).

First choice: Benton CM, Couchman C, Marsden JT, Rees DC, Moniz C, Lim CK. Direct and simultaneous determination of 5-aminolaevulinic acid and porphobilinogen in urine by hydrophilic interaction liquid chromatography-electrospray ionisation/tandem mass spectrometry. Biomed. Chromatogr. 26,1033–1040 (2012).

Reasoning: This was the first rapid, sensitive and selective LC-MS/MS method for the simultaneous detection of 5-aminolaevulinic acid (ALA) and porphobilinogen (PBG), without the need for extensive sample preparation or analyte derivatisation, which has greatly facilitated the clinical analysis of the acute porphyrias. Furthermore, a simple protein precipitation step and centrifugation prior to analysis has also made it perfect for determining plasma/serum concentrations of ALA and PBG.