Spectrofluorometric determination of the antibiotic lasalocid in blood.

A spectrofluorometric assay was developed for the determination of the antibiotic lasalocid in dog blood, based on the intrinsic fluorescence of the compound in ethyl acetate. The assay can measure "total" levels of drug and any metabolites present. The specificity of the assay was verified by TLC separation of the dog blood extract, which indicated the presence of only intact drug. The overall recovery (+/- SD) of lasalocid was 62.0 +/- 3.6% in the concentration range of 1.0-10 mug of compound/ml of dog blood. The sensitivity of the assay is 0.5 mug/ml. The assay was applied to the determination of blood levels of lasalocid in the dog following the intravenous administration of a 5-mg/kg dose.

Effect of lasalocid on the GnRH-induced LH and testosterone release during puberal development in the Brahman bull.

To determine the effect of lasalocid on endocrine patterns associated with puberty, 12 half-sib prepuberal Brahman bulls were allotted by age and weight (174 to 256 d of age; 141 to 243 kg) to control or lasalocid treatments. Bulls in the control treatment were fed a 4:1 corn:cottonseed meal concentrate plus Coastal bermudagrass hay to which the bulls were given ad libitum access. The lasalocid treatment was identical except for the addition of 200 mg of lasalocid.animal-1.d-1. Blood samples were collected frequently before and after GnRH (200 micrograms, i.m.) on d 7, at 28-d intervals thereafter, and within 14 d after puberty (defined as 50 x 10(6) sperm cells with 10% motility). By d 7, bulls fed lasalocid released more LH (P less than .05), but not testosterone (T;P greater than .10), in response to GnRH than controls. At the time that the first sperm cells were observed in an electroejaculate (FS), lasalocid-fed bulls released more (P less than .05) LH and T than controls. At puberty, there was no difference (P greater than .10) between treatments in amount of T released, although lasalocid-fed bulls released more LH (P less than .05). Before puberty, concentrations of LH were positively correlated with concentrations of T in samples collected 1 and 2 h later. Both groups of bulls exhibited a linear increase in T response with advancing age (P less than .005). Release of LH decreased with age in the control bulls (P less than .10) but was unaffected by age in lasalocid-fed bulls. Both groups showed a decreased (P less than .001) LH:T response ratio with advancing age. Results of this study with bulls confirm previous reports in heifers of the enhancing effect of an ionophore on reproductive function.

Determination of bromo-lasalocid in plasma by high-performance liquid chromatography with fluorimetric detection.

A rapid, sensitive, and specific high-performance liquid chromatographic (HPLC) assay was developed for the determination of bromo-lasalocid in plasma. The compound was extracted into isooctane-ethyl acetate (90:10) from plasma saturated with potassium chloride and adjusted to strongly alkaline pH. The residue of this extract was dissolved in methanol-2-methoxyethanol (95:5) and analyzed by HPLC on a 10-micrometer C18 column [mobile phase of methanol-water-2-methoxyethanol-1 M potassium phosphate buffer, pH 3.0 (90:10:2.5:0.2)] using fluorescence detection with excitation at 215 nm and emission at wavelengths greater than 370 nm. The overall recovery of the assay was 65%, with a limit of sensitivity of 0.1 microgram/ml. The method was used to obtain plasma concentration-time profiles in the dog following oral administration of bromo-lasalocid-ethanolate.

Ion transport by lasalocid A across red-blood-cell membranes. A multinuclear NMR study.

Na+ and K+ fluxes mediated by lasalocid A across erythrocyte membranes have been determined from 23Na-NMR peak areas and chemical shifts, respectively. In similar experiments, Cl- transport has been monitored by NMR signal intensities. Taking into account the external pH variations, the results are readily explainable in terms of charge-balance conservation. The effect of disodium 4,4'-diisothiocyanostilbene-2,2'-disulfonate, an anion-exchange inhibitor, has also been studied.