Treatment of wild-type mice with 2,3-butanediol, a urinary biomarker of mice, decreases plasma cholesterol and epididymal fat deposition.

We previously showed that mice exhibit a lean phenotype and slower metabolic ageing. Their characteristics include lower plasma glucose and cholesterol, greater glucose tolerance and insulin sensitivity, and a reduction in age-related weight gain and whole-body fat deposition. In this paper, nuclear magnetic resonance (NMR) spectroscopy-based metabolite analyses of the urine of and wild-type mice identified two isomers of 2,3-butanediol as discriminating urinary biomarkers of mice.

Predicting collision-induced dissociation mass spectra: understanding the role of the mobile proton in small molecule fragmentation.

Rationale: Intramolecular proton migration has been reported to be required for fragmentation by collision-induced dissociation (CID). If the collision energy is required to provide energy for proton movement to a ‘dissociative’ site, it may be possible to predict the optimal collision energy for fragmentation using quantum computational chemistry software. A greater understanding of the mechanism(s) of proton migration is necessary.

Optimization of an automated IS addition system for use in high-throughput quantitative DBS analysis.

Background: Automated DBS direct elution systems are available that incorporate IS spray modules which, unlike conventional IS addition via the extraction solvent, apply IS prior to DBS samples prior to extraction, allowing analyte and IS to be coextracted.

Results: IS spray system parameters were optimized to identify the conditions that produced the best analytical performance in quantitative bioanalytical assays, without interfering with the integrity of the DBS sample prior to extraction.

Conclusion: LC-MS/MS method validations across four representative small molecule assays using the optimized IS spray conditions were demonstrated to produce analytical performance comparable to conventional methods of IS addition, demonstrating that the spray technique is a viable alternative.

DBS direct elution: optimizing performance in high-throughput quantitative LC-MS/MS analysis.

Background: Automated DBS direct elution techniques eliminate the manual extraction burden of DBS bioanalysis, offer good quantitative performance, the ability to eliminate hematocrit-based assay bias, and, previous reports have demonstrated that significant increases in assay sensitivity compared with manual DBS extraction are possible.

Results: An investigation into elucidating parameters for optimized generic DBS direct elution for high sample throughput quantitative bioanalytical applications is presented for the first time. Generic direct elution conditions were identified that enabled LC-MS/MS assay sensitivity to be maximized while retaining acceptable chromatographic performance.

Development and validation of an ultra-performance liquid chromatography quadrupole time of flight mass spectrometry method for rapid quantification of free amino acids in human urine.

An ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-qTOF-MS) method using hydrophilic interaction liquid chromatography was developed and validated for simultaneous quantification of 18 free amino acids in urine with a total acquisition time including the column re-equilibration of less than 18 min per sample. This method involves simple sample preparation steps which consisted of 15 times dilution with acetonitrile to give a final composition of 25 % aqueous and 75 % acetonitrile without the need of any derivatization. The dynamic range for our calibration curve is approximately two orders of magnitude (120-fold from the lowest calibration curve point) with good linearity (r (2) ≥ 0.

How useful is molecular modelling in combination with ion mobility mass spectrometry for 'small molecule' ion mobility collision cross-sections?

Ion mobility mass spectrometry is used to measure the drift-time of an ion. The drift-time of an ion can be used to calculate the collision cross-section (CCS) in travelling wave ion mobility (e.g.

Investigation of different approaches to incorporating internal standard in DBS quantitative bioanalytical workflows and their effect on nullifying hematocrit-based assay bias.

Hematocrit (HCT)-based assay bias (composed of area and recovery bias) is an important contributing factor to the barriers that currently hinder the development and acceptance of dried blood spots (DBS) as a widely used quantitative bioanalytical sampling technique for regulatory studies. This article describes the evaluation of a practical internal standard spray addition technique, used prior to LC-MS/MS analysis, which is demonstrated to nullify the effect of recovery bias. To our knowledge, this is the first time a potential solution to HCT-based recovery bias has been investigated in detail and reported in the literature.

Predicting collision-induced dissociation spectra: semi-empirical calculations as a rapid and effective tool in software-aided mass spectral interpretation.

Rationale: Fifteen molecules were modelled using quantum chemistry, prior to interpreting their collision-induced dissociation (CID) product ion spectra, in a 'blind trial' to establish if calculated protonation-induced bond elongation could be used to predict which bonds cleaved during CID. Bond elongation has the potential to be used as a descriptor predicting bond cleavage.

Methods: The 15 molecules were modelled with respect to protonation-induced bond length changes using Density Functional Theory (DFT).

Construction and physiochemical characterisation of a multi-composite, potential oral vaccine delivery system (VDS).

An increasing human population requires a secure food supply and a cost effective, oral vaccine delivery system for livestock would help facilitate this end. Recombinant antigen adsorbed onto silica beads and coated with myristic acid, was released (∼15% (w/v)) over 24 h at pH 8.8.

Can ion mobility mass spectrometry and density functional theory help elucidate protonation sites in 'small' molecules?

Rationale: Ion mobility spectrometry-mass spectrometry (IMS-MS) offers an opportunity to combine measurements and/or calculations of the collision cross-sections and subsequent mass spectra with computational modelling in order to derive the three-dimensional structure of ions. IMS-MS has previously been reported to separate two components for the compound norfloxacin, explained by protonation on two different sites, enabling the separation of protonated isomers (protomers) using ion mobility with distinguishable tandem mass spectrometric (MS/MS) data. This study reveals further insights into the specific example of norfloxacin and wider implications for ion mobility mass spectrometry.

Understanding collision-induced dissociation of dofetilide: a case study in the application of density functional theory as an aid to mass spectral interpretation.

Fragmentation of molecules under collision-induced dissociation (CID) conditions is not well-understood. This may make interpretation of MSMS spectra difficult and limit the effectiveness of software tools intended to aid mass spectral interpretation. Density Functional Theory (DFT) has been successfully applied to explain the thermodynamics of fragmentation in the gas phase by the modelling the effect that protonation has on the bond lengths (and hence bond strengths).

Tooth extractions in general and due to periodontal reasons in three dental practices: a case-control study.

Objective: Assessment of the risk for tooth extraction in general and due to periodontal reasons in relation to patient factors.

Method And Materials: All patients' charts of three German practices were screened for extractions until 300 patients with extractions (cases) had been collected. All patients without extractions that were screened served as controls.

Commercial 'ready-to-feed' infant foods in the UK: macro-nutrient content and composition.

Quantitative analyses of the macronutrient content of eight popular commercial 'ready-to-feed' baby meals for 6-9-month old infants in the UK market have been undertaken in order to ascertain their nutritional suitability in relation to the total daily dietary intake as well as nutritional profiling of the products. The chemical analyses conducted included Kjeldhal for protein, acid hydrolysis and extraction for fat, phenol sulphuric acid for carbohydrate and Association of Official Analytical Chemists 985.29 for fibre.

Elemental content of commercial 'ready to-feed' poultry and fish based infant foods in the UK.

The study reported herein was conducted in order to establish the concentration of 20 essential and non-essential elements in a representative range of commercial infant foods in the UK targeted for infants aged between 6-12 months. The primary objective of this study was to examine the nutritive values and safety of such complementary infant foods on the UK market in relation to dietary and safety guidelines. Quantitative analyses were conducted on eight different products representing four popular brands (poultry and fish based) of ready to-feed infant foods currently on sale in the UK.

Simultaneous determination of riboflavin and pyridoxine by UHPLC/LC-MS in UK commercial infant meal food products.

An assay for the simultaneous quantitative determination of riboflavin and pyridoxine in eight different complementary infant meal products has been developed in order to (1) estimate the daily intake of these vitamins from commercial infant food consumption, and (2) ascertain their nutritional suitability relative to dietary guidelines for the 6-9 months age group. The method involves mild hydrolysis of the foods, an extraction of the supernatant by centrifugation followed by quantitative determination using ultra-high performance liquid chromatography. Separation of the two water soluble vitamins is achieved within one minute and the resultant sample is also LC-MS compatible.

Ion mobility spectrometry-mass spectrometry (IMS-MS) of small molecules: separating and assigning structures to ions.

The phenomenon of ion mobility (IM), the movement/transport of charged particles under the influence of an electric field, was first observed in the early 20th Century and harnessed later in ion mobility spectrometry (IMS). There have been rapid advances in instrumental design, experimental methods, and theory together with contributions from computational chemistry and gas-phase ion chemistry, which have diversified the range of potential applications of contemporary IMS techniques. Whilst IMS-mass spectrometry (IMS-MS) has recently been recognized for having significant research/applied industrial potential and encompasses multi-/cross-disciplinary areas of science, the applications and impact from decades of research are only now beginning to be utilized for "small molecule" species.

Peptide polarity and the position of arginine as sources of selectivity during positive electrospray ionisation mass spectrometry.

Electrospray ionisation (ESI) is a selective process and, for similar sized analytes, the intrinsic properties of the molecules affect the ionisation process and their response. This research sets out to determine the effect of some of these properties in peptides: peptide polarity and the presence of arginine at positions 1 and 4 in the amino acid sequence on the ESI response. Six peptides; molecular mass ranges 1.

Essential and trace elements content of commercial infant foods in the UK.

There is a paucity of data in respect of the nutritional quality of complementary foods for infants and young children aged between 6 and 12months. The primary objective of this study was to examine nutritive values of such complementary infant food on the UK market in order to ascertain their suitability relative to dietary guidelines for the 6-9months age group. Quantitative analyses were conducted on eight different products representing four popular brands (meat and vegetable based) currently on sale in the UK.

Practical considerations in analysing neuropeptides, calcitonin gene-related peptide and vasoactive intestinal peptide, by nano-electrospray ionisation and quadrupole time-of-flight mass spectrometry: monitoring multiple protonations.

We investigated the pre-electrospray ionisation (pre-ESI) factors; analyte concentration (1-2500  ng/mL), concentration of formic acid (FA) in the mobile phase (0.01, 0.1 and 1%), concentration of the organic modifier (acetonitrile 50-90%) and flow rate (<10  μL/min) on the number of multiple protonations and ESI response for two neuropeptides (of ~3.

Using density functional theory to rationalise the mass spectral fragmentation of maraviroc and its metabolites.

Tandem mass spectrometry (MS/MS) is widely used for the identification of metabolites at all stages of the pharmaceutical discovery and development process. The assignment of ions in the product ion spectra can be time-consuming and hence delay feedback of results that may influence the direction of a project. A deeper understanding of the processes involved in generation of the product ions formed via collision-induced dissociation may allow development of chemically intelligent software to aid spectral interpretation.

"Soft"or "hard" ionisation? Investigation of metastable gas temperature effect on direct analysis in real-time analysis of Voriconazole.

The performance of the direct analysis in real-time (DART) technique was evaluated across a range of metastable gas temperatures for a pharmaceutical compound, Voriconazole, in order to investigate the effect of metastable gas temperature on molecular ion intensity and fragmentation. The DART source has been used to analyse a range of analytes and from a range of matrices including drugs in solid tablet form and preparations, active ingredients in ointment, naturally occurring plant alkaloids, flavours and fragrances, from thin layer chromatography (TLC) plates, melting point tubes and biological matrices including hair, urine and blood. The advantages of this technique include rapid analysis time (as little as 5 s), a reduction in sample preparation requirements, elimination of mobile phase requirement and analysis of samples not typically amenable to atmospheric pressure ionisation (API) techniques.

Effect of increasing concentration of ammonium acetate as an additive in supercritical fluid chromatography using CO2-methanol mobile phase.

The effects of increasing concentrations of ammonium acetate additive in supercritical fluid chromatography were studied on silica, 2-ethyl-pyridine and endcapped 2-ethyl-pyridine stationary phases. The study involved the addition of increasing concentrations of the ammonium acetate either in the mobile phase modifier (methanol) or in the sample solvent. The effects of ammonium acetate on retention and peak shape of the analytes were evaluated.

Can density functional theory (DFT) be used as an aid to a deeper understanding of tandem mass spectrometric fragmentation pathways?

Prediction of tandem mass spectrometric (MS/MS) fragmentation for non-peptidic molecules based on structure is of immense interest to the mass spectrometrist. If a reliable approach to MS/MS prediction could be achieved its impact within the pharmaceutical industry could be immense. Many publications have stressed that the fragmentation of a molecular ion or protonated molecule is a complex process that depends on many parameters, making prediction difficult.