Rapid stereospecific high-performance liquid chromatographic determination of levofloxacin in human plasma and urine.

A rapid high-performance liquid chromatographic (HPLC) method for the determination of levofloxacin in human plasma and urine has been validated. A single-step liquid-liquid extraction procedure was used to isolate levofloxacin from the biological matrix prior to quantitative analysis. The compound was separated on an Inertsil C18 reversed-phase HPLC column and quantified by measuring the UV absorbance at 330 nm. The stereospecificity was achieved in the ligand-exchange mode by incorporating chiral reagents directly into the HPLC mobile phase. Ciprofloxacin was used as the internal standard. The method was linear from 0.08 to 5.18 micrograms ml-1 of levofloxacin in plasma and from 23 to 1464 micrograms ml-1 in urine. The overall utility of the method is reflected in its high sample throughput and easy adaptability to robotic automation, thus making the procedure suitable for pharmacological and pharmacokinetic studies of levofloxacin.

Pharmacokinetics and safety of levofloxacin in patients with human immunodeficiency virus infection.

Levofloxacin, the bacteriologically active isomer of ofloxacin, has microbiologic activity against many pathogens common in human immunodeficiency virus (HIV)-infected patients, including Mycoplasma species which may be cofactors in the progression of HIV disease. The purpose of this phase I, double-blind, randomized (1:1), placebo-controlled trial was to evaluate the pharmacokinetics and safety of levofloxacin hemihydrate in 10 asymptomatic HIV-infected males. Plasma concentrations by chiral high-performance liquid chromatography (HPLC) were evaluated for 48 h after a single 350-mg oral dose, at morning predose during the multiple-dosing phase, and for 72 h at steady state after a week of 350 mg every 8 h orally. Mean +/- standard deviation levofloxacin pharmacokinetic parameters (by noncompartmental moment method) after multiple dosing were as follows: area under the concentration-time curve, 31.24 +/- 5.60 mg.h/liter; apparent total body clearance, 11.18 +/- 1.76 liters/h; renal clearance, 8.63 +/- 2.82 liters/h; steady-state volume of distribution, 104.10 +/- 12.48 liters; and effective half-life, 6.50 +/- 0.51 h. Single-dose parameters were not significantly different from the multiple-dose parameters, with the exception of peak concentrations in plasma, which were 4.79 +/- 1.00 and 6.92 +/- 1.56 mg/liter for single- and multiple-dose data, respectively. Essentially identical parameter values were obtained from curve-fitting analysis when the entire 13-day plasma concentration profiles of the subjects were analyzed simultaneously by a two-compartmental distribution model. Levofloxacin pharmacokinetics in HIV-infected patients remained linear upon multiple dosing. The dosing regimen studied provides levels in plasma and urine well above those found to be effective in vitro against pathogens common in HIV-infected patients. Levofloxacin was well- tolerated in this group of asymptomatic HIV-infected males: there were no statistically significant differences in adverse effects in the two groups (P = 0.22). Use of placebo control helped to differentiate disease-related adverse effects from those related to the study drug.

Bioequivalence of oral and intravenous ofloxacin after multiple-dose administration to healthy male volunteers.

The bioequivalence of oral and intravenous ofloxacin was investigated after the administration of multiple doses of 400 mg every 12 h to 20 healthy male volunteers in a randomized, crossover, open-label study. Ofloxacin concentrations in plasma were evaluated after 4 days of oral or intravenous (1-h infusion) dosing with a 3-day wash-out period between regimens. As expected, delivery to the systemic circulation took slightly longer after the oral dosing (time to maximum concentration of drug in serum of 1.7 h) relative to the 1-h intravenous infusion, but the systemic availabilities of ofloxacin by the two routes of administration were equivalent (area under the concentration-time curve from 0 to 12 h ratio of 95%). Since previous studies have not demonstrated any change in the bioavailability of ofloxacin in infectious disease patients, this study supports the interchangeability of these dosing regimens.

Bioavailability and pharmacokinetics of oral ofloxacin formulations in normal subjects.

The relative bioavailability and pharmacokinetics of ofloxacin tablets and a reference oral solution of ofloxacin were compared in 32 normal male subjects using a randomized two-way crossover design. After an overnight fast, subjects were randomized to receive a single 200 mg or 300 mg dose of ofloxacin (tablet or solution) and blood samples were obtained prior to and 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 16, 24, 36, 48, 72, 96, and 120 hours after the dose. After a 5-day wash-out period, subjects were administered the same dose but of the other formulation, and blood samples were collected in an identical manner. Plasma concentrations of ofloxacin were determined by high-pressure liquid chromatography. The results showed that ofloxacin tablets were more slowly absorbed when compared to the solution and mean peak plasma concentrations were obtained in about 1.5 hours for the tablet preparation. Maximum plasma concentrations were higher after administration of the solution (Cmax = 2.24 micrograms/ml, 200 mg; Cmax = 3.25 micrograms/ml, 300 mg) compared to the tablet (Cmax = 1.74 micrograms/ml, 200 mg; 2.61 micrograms/ml, 300 mg). The bioavailability of ofloxacin tablets was greater than 98% compared to the solution. The other pharmacokinetic parameters were similar between the two dosage formulations. Ofloxacin tablets revealed an apparent volume of distribution of 1.5 l/kg, an elimination half-life of 5.6 hours, and a total clearance of 251 ml/min. In addition, a linear increase in plasma concentrations was observed when the dose of ofloxacin was increased. In summary, ofloxacin tablets was found to be reliably bioavailable and bioequivalent to the reference solution.

The effect of food or milk on the absorption kinetics of ofloxacin.

We have studied the effects of food or milk on the absorption of ofloxacin in 21 healthy male volunteers in a three-way crossover design. Milk did not alter the rate or extent of absorption of ofloxacin or its elimination. Food altered the onset and/or rate of absorption, but not the extent of absorption or the elimination rate. Thus, food reduced peak ofloxacin concentrations (Cmax) by 20% compared with fasting conditions and the time to reach maximum concentration (tmax) was prolonged on average by 1 h. However, the extent of absorption and the half-life (t 1/2) of ofloxacin were the same after each treatment. These data indicate that food and milk have a clinically insignificant effect on ofloxacin absorption.

Safety of multiple doses of ofloxacin in healthy volunteers.

A safety profile of ofloxacin, a new fluoroquinolone antibiotic, was assessed in twelve healthy male volunteers. Ofloxacin was dosed at 400 mg twice daily over a 10-day course of treatment. This evaluation included chemistry and haematologic profiles, neurologic evaluations, ophthalmologic examinations, audiometric testing, and electrocardiogram before, during and the end of the study. In addition, plasma samples were analysed for ofloxacin prior to each morning dose and a complete plasma concentration profile was performed on day 6. Gastrointestinal complaints such as nausea, diarrhoea and upset were the most frequent side effects reported. The only other adverse event that occurred in more than one volunteer was headache. None of the subjects in this study developed a serious side effect and no physical or psychological changes were observed. Laboratory values remained in their normal ranges with the exception of one volunteer who developed a 3-fold rise in one liver function test at the end of the study. The subject was asymptomatic and this aberration resolved without sequelae. In summary, a 10-day course of ofloxacin was found to be safe and have a low potential for toxicity. The side effects observed in this study were mild and similar to those observed with other fluoroquinolones.

A sensitive high performance liquid chromatography microassay for bemoradan, a novel cardiotonic agent, in plasma/serum using a semi-automatic liquid/solid extraction sample preparation system--AASP.

A high performance liquid chromatographic method for the measurement of bemoradan levels in plasma/serum is described. This method uses Varian's AASP (Varian Associates, Sunnyvale, CA, USA), a semi-automatic liquid/solid extraction sample preparation system. It requires only small volumes of plasma/serum samples (0.2-1 mL) and needs no organic solvent for sample preparation. The mean recovery of bemoradan at plasma or serum concentrations of 0.5-100 ng/mL is 82%. The assay has a detection limit of 0.5 ng/mL (when 1 mL of plasma/serum is used) and is linear in the concentration range 0.5-500 ng/mL.

Determination of the calcium antagonist flunarizine in biological fluids by gas-liquid chromatography.

A method is described for the quantitation of flunarizine in biological fluids including plasma, urine, milk, fecal and tissue homogenates using the analogue cinnarizine as the internal standard. As little as 1.5 ng of flunarizine per ml of plasma can be accurately quantitated, this being achieved by the combination of a selective extraction procedure and a nitrogen detector. The method has been used to determine the concentration of flunarizine in biological fluids in support of human and animal pharmacokinetic studies.