Noninvasive in vivo 13C-NMR spectroscopy in the rat to study the pharmacokinetics of 13C-labeled xenobiotics.

Noninvasive NMR methodology has been developed to enable monitoring of 13C-labeled xenobiotics in the rat in vivo. 2,2-Dichloro-1-(2-chlorophenyl)-1-(4-chlorophenyl)-[3-13C]-propane can be detected in the liver of intact rats by in vivo 13C surface coil NMR spectroscopy after ip administration of the compound. The experiments were performed at 1.9 and 9.4 Tesla. The intrahepatic changes of the signal intensity of the labeled compound were followed as a function of time. In the days following administration, the concentration decreased and dropped to values below the detection limit after 12 days. The study demonstrates the feasibility of studies on pharmacokinetics of 13C-labeled compounds in the rat using noninvasive, in vivo surface coil NMR spectroscopy in animals. The sensitivity allows the detection of a single dose of the drug of 200 mg/kg, but can be improved.

Proton relaxation enhancement by means of serum albumin and poly-L-lysine labeled with DTPA-Gd3+: relaxivities as a function of molecular weight and conjugation efficiency.

A series of poly-L-lysine chains, with molecular weight ranging from 3300 up to 102,000 Da, were labeled with DTPA-Gd3+. No significant differences in longitudinal and transversal relaxivity, could be demonstrated as a function of the chain length. The R1 and R2 relaxivities were respectively 2.5 and 5 times superior to those of plain DTPA-Gd3+ (at 2.4 T). Bovine serum albumin was also labeled in a way that a wide (DTPA-Gd3+)/BSA range (3-39) was obtained. The longitudinal relaxivity, of these paramagnetically labeled albumins, increased with increasing (DTPA-Gd3+)/BSA ratios. This effect was most pronounced at very low (DTPA-Gd3+)/BSA ratios.

Rapid method for the analysis of red blood cell fatty acids by reversed-phase high-performance liquid chromatography.

A high-performance liquid chromatographic method was developed for a rapid qualitative and quantitative analysis of p-bromophenacyl esters of red blood cell fatty acids in humans. Both free and bound fatty acids, extracted with hexane-2-propanol (3:2) from packed red blood cells were derivatized with p-bromophenacyl bromide and analysed. Ten identical samples taken from a mixed pool of packed red blood cells from healthy subjects were analysed on two different columns. The fatty acid p-bromophenacyl esters were analysed on a 10 RP-18 column with methanol-acetonitrile-0.01 M ammonium formate as mobile phase and also on a 10 RP-8 column with acetonitrile-0.01 M ammonium formate as mobile phase. The two methods gave analogous results except in total analysis time: that on a 10 RP-8 column is ca. 40% shorter. Furthermore, a quantitative analysis of a standard solution to evaluate the extraction procedure in the absence or in the presence of the red blood cell core indicated a significant difference when the core is present.

Direct liquid introduction LC/MS microbore experiments for the analysis of nucleoside material present in human urine.

In order to obtain detection limits low enough for the analysis of nucleoside material in biological samples, a direct liquid introduction (liquid chromatographic/mass spectrometric DLI) system was upgraded by inserting a self-built desolvation chamber between the DLI probe and the ion source and by switching to microbore high-performance liquid chromatography (HPLC) on a C-18 column. The system was evaluated for the analysis of pure nucleoside and 2'-deoxynucleoside compounds in CH3OH + H2O (80/20) and for the analysis of nucleoside mixtures which were separated using 0.01 M ammonium formate + methanol (97:3) as eluant. The detection of structurally important fragment ions in the lower mass region which could not be observed before and an enhanced sensitivity are considered to be the main improvements. In combination with an appropriate clean-up procedure the system was evaluated for the DLI liquid chromatographic/mass spectrometric analysis of some nucleosides present in a human urine sample. Pseudouridine and 5,6-dihydrouridine were detected and identified.

Plant antiviral agents. III. Isolation of alkaloids from Clivia miniata Regel (Amaryllidaceae).

Lycorine (1), one of the main alkaloids of the Amaryllidaceae family, was found to be responsible for the pronounced antiviral activity of the crude extracts from the roots and leaves of Clivia miniata Regel. It was shown that the inhibitory activity of lycorine on the cytopathogenic effect caused by a DNA and several RNA viruses on VERO cells was not virucidal. Poliomyelitis virus inhibition occurred at lycorine concentrations as low as 1 microgram/ml, whereas concentrations exceeding 25 micrograms/ml were found to be cytotoxic. Clivimine (2), clivonine (3) and cliviamartine (4) were also isolated and characterized, but these alkaloids exhibited no antiviral properties.

Isolation and identification of 2-oxo-5-guanidinovaleric acid in urine of patients with hyperargininaemia by chromatography and gas chromatography/mass spectrometry.

An unknown guanidino-positive peak has been identified in urines of three sisters affected with hyperargininaemia. Identification was made on the basis of its similarity with the liquid and thin-layer chromatographic characteristics of enzymatically synthesized 2-oxo-5-guanidinovaleric acid. Identification was also made by combined gas chromatography-mass spectrometry of the unknown compound peak. The synthesis of enzymatically formed 2-oxo-5-guanidinovaleric acid was controlled by nuclear magnetic resonance and combined gas chromatography-mass spectrometry.

Reduction of cycloalkanones by Aspergillus niger.

The mould Aspergillus niger was grown in a carefully established culture medium containing one of the following test substrates: cyclopentanone, -hexanone, -heptanone, and their 2-substituted methyl derivatives. Growth curves, glucose consumption curves and curves showing the course of the oxido-reduction reaction are given.