Test strip enzyme immunoassays and the fast screening of nortestosterone and clenbuterol residues in urine samples at the parts per billion level

The preliminary results of an investigation into the development of "on-site" test strip enzyme immunoassays for the screening of urine samples for the presence of growth promoters, such as 17 beta, 19-nortestosterone and clenbuterol at the parts per billion level are described. Urine samples, enzyme-labelled analyte and a nitrocellulose test strip, containing immobilized antibodies, are incubated together, after which the strip is placed in a chromogen-containing substrate solution for colour reaction. Using prefabricated strips, the tests can be performed in 45-60 min. A similar assay was worked out using a dot-blotting device, allowing the test to be performed in 20-50 min. The tests are simple and easy to perform outside the laboratory. Urine samples identified positive by gas chromatography mass spectrometry were also found to be positive with these test strips and, so far, no false-positive results have been encountered. With standard additions to blank urine samples, positive samples could be distinguished above the 5 ng ml level. However, samples from treated calves contain one or more metabolites of the parent compound, which increase the sensitivity of the assays. Although the tests described can be improved and still have to be evaluated further by analysing more urine samples, the preliminary results are very promising and give a lead to further research into the applicability of such "on-site" tests in residue analysis.     

Mass spectral characterization of C-glycosidic flavonoids isolated from a medicinal plant (Passiflora incarnata)

The four major C-glycosidic flavonoids isolated from Passiflora incarnata were identified as schaftoside, isoschaftoside, isovetexin-2'-O-glucopyranoside and isoorientin-2'-O-glucopyranoside on the basis of mass spectral and 13C NMR data. The daughter ion spectra of [M + H]+ ions of schaftoside and isoschaftoside showed differences for the [M + H - 104]+ ions, which could be rationalized by hydrogen bonding effects. In the negative-ion mode, pronounced differences were found for the [M - H - 90]- and [M - H - 120]- ions, formed by prevalent fragmentation in the C-6-linked sugar moiety. With respect to isovitexin-2'-O-beta-glucopyranoside and isoorientin-2'-O-beta-glucopyranoside, the daughter ion spectra of both the [M + H]+ and [M - H]- ions provided evidence for a 1----2 linkage in the diglucosidic moiety. Support for C-6 glucosylation was obtained by recording the daughter ion spectra of [M - H - 162]- ions, which were in good agreement with that obtained for [M - H]- ions of isovitexin.     

Primary sequences of insulin-like growth factors 1 and 2 isolated from porcine plasma

Insulin-like growth factors 1 and 2 were purified from porcine plasma. In addition to the determination of their isoelectric points, the primary structures of both proteins were determined, using low microgram quantities of protein, by the versatile combination of time-of-flight plasma desorption mass spectrometry and automated Edman degradation. Porcine insulin-like growth factor 1 was shown to be homologous to both human and bovine proteins; the type 2 growth factor showed one mutation to both human and bovine type 2 proteins.     

Mechanisms of repeller-induced effects in thermospray liquid chromatography/mass spectrometry

The application of a high potential at the repeller electrode, positioned opposite to the sampling cone in order to increase the sampling efficiency, can induce fragmentation in thermospray mass Spectrometry. Until now, this fragmentation has been attributed to collision-induced dissociation. As a result of studies on the changes in the reagent gas composition in the thermospray buffer ionization mode as a function of the repeller potential in the positive-ion mode, it appears that three different processes are occurring. At low repeller potentials, the thermospray mass spectra of the eluent are determined by the proton affinities and the concentrations of the various solvent constituents, and the stabilities of the formed cluster ions under the ion source conditions. With an increase in the repeller potential, collision-induced dissociation of the background ions starts to occur. When the kinetic energy of the ions and cluster ions becomes high enough, endothermic proton transfer and solvent-switching reaction pathways are opened. For the relatively volatile analytes studied, e.g. aniline, acetophenone, benzaldehyde and benzoic acid, similar effects are observed.     

Metallic susceptibility in giant molecule: Pd561Phen36O200

The giant metal cluster molecule Pd₅₆₁Phen₃₆O _x is the largest member of a series that has recently been prepared by Schmid [1]. Pronounced size effects are expected to show up in the magnetic properties of metal particles of the size of the Pd-561 core of the molecule. We have measured the susceptibility on four different batches of the material, with quite satisfactory reproducibility. There is a low temperature tail that is attributed to small amounts of impurities. The nearly temperature-independent paramagnetism that is observed is most probably due to an exchange enhanced Pauli spin susceptibility and is already close to the bulk value. The absence of size effects, i.e. a drop at low temperatures, is either due to interactions between the clusters or a result of a very small energy level spacing.     

Gold and platinum molecular cluster compounds studied by 197Au Mössbauer Spectroscopy

The technique of ¹⁹⁷Au Mössbauer absorption spectroscopy has been used to study four varieties of the high nuclearity gold molecular cluster compounds abbreviated as Au₅₅, all having a cuboctahedral structure, with 12 PPh₃-ligands or modifications of PPh₃. The technique of emission spectroscopy developed in our laboratory [1] has been applied to four different molecular platinum carbonyl cluster compounds of varying cluster nuclearity, abbreviated as Pt₃₈, Pt₂₆, Pt₂₄, and Pt₁₉ [2]. The four Au55 compounds were studied, both as dry materials and as frozen solutions. The M6ssbauer parameters of the chlorine ligated site of the water soluble version differ strongly due to the effect of Na⁺ on the Au-Cl distances. In the frozen solution, the effect of the ionic charge on this cluster can be clearly seen. The similarity of the spectra of all the Pt compounds, especially the occurrence of a substantial singlet contribution in all spectra, is explained by a coalescing mechanism for the low-nuclearity clusters, induced by the neutron irradiation damage inflicted while preparing the sample as M6ssbauer sources. The observed decrease of the absorption intensity with increasing temperature is evidence for a cluster character of the sample remaining after neutron irradiation.     

Deactivation of silica surfaces with a silanol-terminated polysiloxane; structural sharacterization by inverse gas chromatography and solid-state NMR

Retention gape deactivated with Silicone OV-1701-OH show good chromatographic performance and remarkable stability against water induced stationary phase degradrdation. In an attempt to better understand the findamentals off the deactivation process using silanol terminated polysiloxanes, a fumed silica was deactivated with Silicon OV-1701-OH. In contrast to fused silic capillaries, fumed silica (Aerosil A-200) can be studied by ²⁹Si cross-polarization magic-angle-spinning (CPMAS) NMR, thus serving as a model substrate for fused silica. Retention data from inverse gas chromatography at infinite dilurion and ²⁹Si CP MAS NMR data of five Aerosil phases, differing in residual silanol surface concentration, are correlated with the aim of validating this approach for stationary phase characterization. A comparatively detailed model of the deactivating polymer layer that explains the observed absorption activities is deduced. Surface silanols are shown to play a key role in the polymer layer, the structure of which is of primary importance for the absorption behavior after deactivation. Contrary to common belief, the absolute silanol surface concentration after deativation is only of secondary importance for the overall absorption activity. High silanol surface concentrations enhance degradation of the polysiloxane chains into small cyclic fragments as well as subsequent absorption and immobolization to the silica substrate surface. The mobility of linear polysiloxane chains in the kHz regime (as determined bby NMR cross-polarization dynamics) appears to determine the extent which the residual silanols are accessible for analytes. It is therefore anticipated that there is an optimum silanol surface concentration of fused silica surfaces to be deactivated with silanol terminated polysiloxanes; it should be lazrge enough to adsord polymer fragments, but not large to avoid excessive residual silanol activity.     

Quercetin Prevents UV-lnduced Local Immunosuppression, but Does Not Affect UV-lnduced Tumor Growth in SKH-1 Hairless Mice

Abstract— Ultraviolet is thought to induce skin tumors by its dual activity as a mutagenic agent and a suppressor of cell-mediated immunity. In the present study the effects of quercetin, a flavonoid-containing compound, on carcinogenesis and immunosuppression were studied in SKH hairless mice exposed to suberythemal doses of UV for up to 17 weeks. It was found that quercetin did not affect the onset or growth of non-melanoma skin tumors resulting from UV exposure. In contrast, it prevented the suppression in contact hypersensitivity (CHS) to picryl chloride induced by UV. The mechanism of this prevention might be explained by the observation that the decreased number of epidermal Langerhans'cells is partly prevented by the quercetin. Quercetin did not alter the effects of UV in increasing numbers of spleen and lymph node cells, only partly in decreasing the CD8-positive cells in spleen cell populations and decreasing the lym-phoproliferative response of spleen cells to the mitogens concanavalin A and phytohemagglutinin. Thus oral quercetin did not prevent UV-induced carcinogenesis although it restored the skin-associated CHS response probably by protecting the antigen-presenting cells in the skin.