Anna Przyborowska

Deconvolution gas chromatography/mass spectrometry of urinary organic acids – potential for pattern recognition and automated identification of metabolic disorders

The National Institute of Standards and Technology (NIST) Automated Mass Spectral Deconvolution and Identification System (AMDIS) is applied to a selection of data files obtained from the gas chromatography/mass spectrometry (GC/MS) analysis of urinary organic acids. Mass spectra obtained after deconvolution are compared with a special user library containing both the mass spectra and retention indices of ethoxime-trimethylsilyl (EO-TMS) derivatives of a set of organic acids. Efficient identification of components is achieved and the potential of the procedure for automated diagnosis of inborn errors of metabolism and for related research is demonstrated.

A library of atmospheric pressure ionization daughter ion mass spectra based on wideband excitation in an ion trap mass spectrometer

A searchable library of MS/MS spectra obtained using a quadrupole ion trap mass spectrometer and electrospray or atmospheric pressure chemical ionization is presented. The application of wideband excitation (activation) and normalized collision energy leads to highly reproducible mass spectra which are searched using the NIST algorithm. Flow injection and LC/MS/MS applications of this powerful technique in the biomedical (diastereoisomeric steroids, morphine glucuronides, isovalerylcarnitine) and environmental (pirimicarb and desmethyl-pirimicarb) areas are described.

Isatin, regional distribution in rat brain and tissues

Isatin has recently been identified in rat tissues and normal human urine, where it forms the major proportion of the endogenous monoamine oxidase inhibitor, tribulin. In this paper, we show that isatin, measured by gas chromatography/mass spectrometry, has a distinct regional distribution in rat tissues, with highest concentrations in seminal vesicles (1.6 ?g/g) and vas deferens (3.4 ?g/g). There was also a discontinuous distribution within rat brain, concentrations being highest in the hippocampus (0.13 ?g/g).

The effect of Khat (Catha edulis) as an appetite suppressant is independent of ghrelin and PYY secretion

BACKGROUND The leaves of the khat plant (Catha edulis) are chewed for their pleasurable effects. Chewing releases cathinone which may decrease appetite through an unknown mechanism. Levels of the peptide ghrelin increase with hunger and decrease immediately post-prandially, while peptide YY is released following a meal. We hypothesised that the anorexigenic effects of khat may be mediated through changes in these hormones. MATERIALS AND METHODS Six habitual khat chewers attended on two separate occasions. For a period of 3h they chewed either khat leaves or lettuce. Blood pressure (BP) and pulse rate (PR) were monitored throughout, as were subjective assessments of hunger and fullness. Plasma samples were analysed for cathinone, ghrelin and PYY levels. RESULTS Chewing khat significantly decreased subjective feelings of hunger and increased fullness (p<0.05) but had no effect on ghrelin and PYY levels. Khat led to an increase in cathinone levels as well as an increase in BP and PR. Cathinone levels correlated positively with fullness and pulse rate and negatively with hunger. CONCLUSIONS Chewing khat decreases subjective feelings of hunger and increases systemic sympathetic tone, but has no effect on ghrelin and PYY levels. We conclude that the anorexigenic effect of khat may be secondary to central mechanisms mediated via cathinone.

Urinary but Not Brain Isatin Levels Are Reduced in Germ‐Free Rats

Germ‐free rats excreted considerably smaller amounts of the monoamine oxidase‐inhibiting compound isatin than the substantially larger output by conventional animals of the same strain, although concentrations in brain and other tissues were similar in the two groups. Thus, isatin is likely to be elaborated both endogenously in rat tissues and “exogenously” by flora inhabiting the lumen of the alimentary tract.

Isatin (indole-2,3-dione) in urine and tissues: Detection and determination by gas chromatography—mass spectrometry

A simple procedure based upon capillary column gas chromatography-mass spectrometry (GC-MS) is described for the detection and determination of isatin (indole-2,3-dione) in body fluids and tissues. After addition of 5-methylisatin as internal standard to urine or tissue homogenates, organic extracts are dried and derivatized successively with hydroxylamine hydrochloride and the reagent N-tert.-butyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA). The tert.-butyldimethylsilyl derivatives obtained show good GC-MS properties and allow quantification by selected-ion monitoring of m/z 333 (isatin) and m/z 347 (internal standard). Adult and newborn human urine output values lie in the ranges 0.4-3.2 mg/mmol of creatinine (5-30 mg per 24 h) and 0.002-0.518 mg/mmol of creatinine, respectively. There is a discontinuous regional distribution in rat tissues. The GC-MS properties of a number of derivatives formed by successive reaction of isatin with hydroxylamine hydrochloride (or methoxyaminehydrochloride or ethoxyamine hydrochloride) and MTBSTFA, bis(trimethylsiyl)trifluoroacetamide, pentafluoropropionic anhydride or pentafluorobenzyl bromide are also described.