Ra Dezeeuw

Muscarinic M2 receptors in bovine tracheal smooth muscle: discrepancies between binding and function

Previous work showing that AF-DX 116, a cardioselective muscarinic antagonist in functional experiments, does not discriminate between muscarinic receptors in bovine cardiac and tracheal membranes has been extended. In addition to AF-DX 116 we used the muscarinic antagonists, atropine, pirenzepine, 4-DAMP methobromide, gallamine, hexahydrosiladifenidol and methoctramine, in radioligand binding experiments on bovine cardiac left ventricular and tracheal smooth muscle membranes. The functional antagonism of the methacholine-induced contraction of bovine tracheal smooth muscle strips was also evaluated. An excellent correlation was found for all compounds between the binding affinities for muscarinic receptors in cardiac and tracheal smooth muscle membranes; moreover, the affinities found in cardiac membranes correspond with the pA2 values reported for atrial preparations of rat and guinea pig. However, significant and occasionally marked discrepancies were found between binding and functional affinities of these muscarinic antagonists on bovine tracheal smooth muscle.

A Single-Column Procedure on Bond Elut Certify for Systematic Toxicological Analysis of Drugs in Plasma and Urine

A single-column solid-phase extraction procedure was developed for the screening of acidic, neutral, and basic drugs from plasma. The recoveries of all 25 tested drugs exceeded 82%. After the plasma had been diluted with phosphate buffer (pH 6.0), the drugs were extracted using a single Bond Elut Certify column. The acidic and most of the neutral drugs were eluted by acetone/chloroform (1:1) and the basic drugs were eluted by 2% ammoniated ethyl acetate. Some neutral drugs appeared in both fractions. The two fractions were collected separately and evaporated until approximately 100 microL of solvent remained in the tube. Both fractions were analyzed separately on a gas chromatograph equipped with a wide-bore capillary column and a flame ionization detector. The procedure could also be used for urine samples.

The M2 selective antagonist AF-DX 116 shows high affinity for muscarine receptors in bovine tracheal membranes

SummaryWe have characterized the muscarine receptors in bovine tracheal and left ventricular membranes using 3H-dexetimide/pirenzepine and 3H-dexetimide/AF-DX 116 competition studies. Pirenzepinc exhibited low (M2) affinity binding to both preparations; Kd was 590 nM in left ventricle and 463 nM in trachea. AF-DX 116 exhibited high (M2) affinity binding to left ventricle (Kd = 95.6 nM); in tracheal membranes it bound with high (M2) affinity (Kd = 40.7 nM) to 74% of the receptors and with low (M3) affinity (Kd = 2.26 μM) to 26% of the receptors. It is concluded that bovine tracheal muscle membranes contain a heterogeneous population of muscarine binding sites, the majority having M2 (heart) subtype characteristics and being located on the smooth muscle membranes; a minority having M3 (exocrine gland) subtype characteristics and presumed to be located in submucosal glands. This is the first report of high affinity binding of AF-DX 116 to non-cardiac peripheral muscarine receptors.

Rapid and sensitive liquid chromatographic determination of carbamazepine suitable for use in monitoring multiple-drug anticonvulsant therapy.

A rapid, sensitive and selective method for the determination of carbamazepine and its major metabolite in plasma has been developed. Other commonly used anticonvulsants can be determined in the same procedure without interference. After extraction with dichloromethane, the components are separated by high-pressure liquid chromatography without further clean-up or concentration on a column packed with small-particle silica gel. The mean recovery from plasma is 98.6% with a relative standard deviation of 1.6%. The detection limit for carbamazepine is approximately 2 ng/ml, requiring 1 ml of plasma.

A study of the separation of enantiomers of some aromatic carboxylic acids by high-performance liquid chromatography on a β-cyclodextrin-bonded stationary phase

Abstract The enantiomers of some aromatic carboxylic acids of biomedical interest were separated by high-performance liquid chromatography on a β-cyclodextin-bonded stationary phase. Evaluation of the system shows that the structure of the acids plays an important role in the possibility for separation of the enantiomers. Moreover, the pH as well as the buffer components of the mobile phase have a pronounced effect on the retention times and the selectivity of the system. In order to reduce tailing, the column temperature was increased; this resulted in better peak resolution. Anomalous behaviour was seen in the relation between retention time and sample concentration. At low amounts of injected sample the retention times increased, but this could not be ascribed to adsorption processes. This phenomenon caused great problems in the quantitative analysis of the two enantiomers with this system.

COMPARISON OF 2 BETA-CYCLODEXTRIN BONDED STATIONARY PHASES FOR HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY - ELUTION ORDER AND OPTICAL PURITY OF ENANTIOMERS OF CYCLOHEXYLPHENYLGLYCOLIC ACID

Comparaison entre colonne du commerce et du laboratoire, et entre mesure de purete optique par chromatographie ou calorimetre differentielle a balayage

A RAPID GAS-CHROMATOGRAPHIC METHOD FOR THE FINGERPRINTING OF ILLICIT COCAINE SAMPLES

A gas chromatographic (GC) fingerprint method, based on the presence or absence of six congeners, was developed for illicit cocaine samples. The fingerprint utilizes the relative abundances of these congeners towards each other, disregarding cocaine as the main constituent, and can be expressed numerically or graphically in the form of pictograms for rapid visual comparison. The method can be applied directly to a solution of the sample in chloroform, without previous workup procedures. More than 70 unrelated samples were analyzed and a great variation was observed in the parameter composition. On the other hand, a remarkable similarity could be seen between related samples. The GC fingerprint method may be considered an important contribution for sample comparison, as is exemplified by a subdivision of the analyzed samples in different categories, based on the number and types of congeners found.

The metabolic fate of the dopamine agonist 2-(N-propyl-N-2-thienylethylamino)-5-hydroxytetralin in rats after intravenous and oral administration II. Isolation and identification of metabolites

1. The in vivo metabolic pathways of 2-(N-propyl-N-2-thienylethylamino)-5-hydroxytetralin (I) in rats have been established, using in vitro metabolism as a complementary technique. 2. All identified metabolites were conjugates. Glucuronidation at the phenolic group yields the major metabolite, accounting for 50% (i.v.) or 65% (oral) of the dose. The corresponding sulphate conjugate of I is virtually absent (less than 0.2% dose). 3. Hydroxylation of I, at the ortho position to the phenolic hydroxy group, yields 6-hydroxy-I (II), accounting for about 13% (i.v.) or 9% (oral) dose. This catechol is excreted, as a glucuronide, almost exclusively into the bile. Both the 5- and the 6-glucuronide of II were detected in about equal amounts. 4. Metabolism of I in vitro showed that under oxidative conditions, depropylation of I occurred. Conjugation of 3H-I in the presence of UDPGA or PAPS, was successful in yielding the glucuronide and sulphate conjugates.

DETERMINATION OF CARBOXYLIC-ACIDS IN PICOMOLE RANGE AFTER DERIVATIZATION WITH PENTAFLUOROBENZYL BROMIDE AND ELECTRON-CAPTURE GAS-CHROMATOGRAPHY

Abstract A method is presented for the determination of picomole quantities of carboxylic acids by gas chromatography in combination with electron capture detection. The acids are extracted from aqueous media into dichloromethane by ion-pair extraction with tetrapentylammonium ions, and derivatized as their pentafluorobenzyl esters. These derivatives have good chomatographic properties with minimum detectable amounts of ca . 0.15 pg 250 pg or greater quantities of the acids can be used. Recoveries are ca . 90% with a precision of ca . 6% at the 10-ng level.

INTERLABORATORY APPLICABILITY OF A RETENTION INDEX LIBRARY OF DRUGS FOR SCREENING BY REVERSED-PHASE HPLC IN SYSTEMATIC TOXICOLOGICAL ANALYSIS

Abstract The retention indices (RI) of 47 selected acidic, neutral and basic drugs were determined on 7 reversed-phase (octyl- and octadecylsilica) columns in two laboratories in the 1-nitroalkane scale, using either 1-nitroalkane homologues or selected drugs, whose RI values were previously determined on the reference column. Obtained values were compared with the library values, determined previously on the reference column. Retention indices, calculated with drugs as RI markers, showed distinctly lower deviations from the library values and lower inter-column variability: The mean standard deviation of RI for all drugs analyzed on all columns in the 1-nitroalkane scale was 44.3 RI units, against 10.3 units when selected drugs were used as RI markers. The deviations from the listed values, calculated for each column separately with drugs as markers, were in 95% of cases smaller than 20 RI units, and in 80 % smaller than 10 units. The largest differences between the experimental and listed values were ob...