Simultaneous determination of rifampicin and sulbactam in mouse plasma by high-performance liquid chromatography.

A simple and rapid high-performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed and validated for the simultaneous determination of rifampicin and sulbactam in mouse plasma. Plasma samples were deproteinized with acetonitrile and separated by HPLC on a RP-18 (125 x 4 mm, 5 microm) column and gradient elution with potassium dihydrogen phosphate solution (pH 4.5; 50 mm) and acetonitrile at a flow-rate of 1.0 mL/min. Rifampicin and sulbactam were monitored at 230 nm and confirmed by means of their UV spectra using a diode-array detector. The method was linear at plasma levels from 1 to 100 microg/mL for rifampicin and from 5 to 200 microg/mL for sulbactam. The limits of quantification were 0.6 microg/mL for rifampicin and 4.2 microg/mL for sulbactam. The intra- and inter-day precisions of the method (RSD) were lower than 5% for both compounds. Average recoveries of rifampicin and sulbactam from mice plasma were 98.2 and 89.3%, respectively. The developed method was successfully applied to the determination of the pharmacokinetic profile of both compounds in mice.

Electrochemical study of imipenem's primary metabolite at the mercury electrode. Voltammetric determination in urine.

Imipenem shows a fast chemical conversion to the more stable imin form (identical to that from biochemical dehydropeptidase degradation) in aqueous solutions that shows a wave at lower cathodic potential than the imipenem one. The aim of this work is the study of the electrochemical behaviour of the primary metabolite of imipenem (M1) and the proposal of electrochemical methods for the determination of M1 in human urine samples. Electrochemical studies were realized in phosphate buffer solutions over pH range 2.0-8.0 using differential pulse polarography, dc-tast polarography, cyclic voltammetry and linear sweep voltammetry (staircase). In acidic media, a non-reversible diffusion-controlled reduction involving two electrons and two protons occurs and the mechanism for the reduction was suggested. A differential pulse polarographic method for the determination of M1 in the concentration range 10(-6) to 10(-4)M with a detection limit of 4.5 x 10(-7)M was proposed. Also, a method based on controlled adsorptive pre-concentration of M1 on the hanging mercury drop electrode (HMDE) followed by linear sweep voltammetry allows its determination in the concentration range 2 x 10(-9) to 4 x 10(-8)M with a detection limit of 1.05 x 10(-9)M. The proposed methods have been used for the direct determination of M1 in spiked human urine and real human-derived urine with good results and should be appropriate for monitoring purposes.

Metal toxicity in Chlamydomonas reinhardtii. Effect on sulfate and nitrate assimilation.

Cadmium (Cd(2+)) or copper (Cu(2+)) ions are toxic for Chlamydomonas reinhardtii growth, at 300 microM, and the alga may accumulate about 0.90+/-0.02 and 0.64+/-0.02% of its dry weight, respectively. Metal contamination changes the elemental composition of dried alga biomass, which indicates the possibility to use C. reinhardtii as biosensor and bioremediator of the aquatic contamination by heavy metals. Either, Cd(2+) or Cu(2+), inhibits about 20% of the nitrate consumption rate by the cells, while only Cd(2+) increases about 40% the sulfate consumption rate. The presence of 1 mM calcium (Ca(2+)) in the culture medium increases the C. reinhardtii productivity (about 50%), the nitrate uptake rate (about 20%) and the sulfate uptake rate (about 30%). In addition, Ca(2+) overcomes the Cd(2+) (300 microM) toxicity by decreasing (about 35%) the intracellular accumulation of metal. Sulfur-starvation induces in C. reinhardtii the expression of serine acetyltransferase and O-acetylserine(thiol)lyase activities, but decreases 50% the consumption rate of nitrate by the cells. Sulfate is also required for the full expression of the nitrate reductase (NR), nitrite reductase (NiR) and glutamate synthase activities.

Electrochemical reduction of cefminox at the mercury electrode and its voltammetric determination in urine.

The electrochemical behaviour of cefminox in phosphate buffers solutions over pH range 2.0-9.0 using differential-pulse polarography, DC-tast polarography, cyclic voltammetry and linear sweep voltammetry (staircase) has been studied. In acidic media, a non reversible diffusion-controlled reduction involving two electrons occurs and the mechanism for the reduction was suggested. A differential-pulse polarographic method for the determination of cefminox in the concentration range 5.8x10(-6)-6.0x10(-5) M with a detection limit of 1.76x10(-6) M was proposed. Also, a method based on controlled adsorptive pre-concentration of cefminox on the hanging mercury drop electrode followed by linear sweep voltammetry, allows its determination in the concentration range 8.3x10(-8)-1.5x10(-6) M with a detection limit of 2.47x10(-8) M. These methods have been used for the direct determination of cefminox in human urine with recoveries between 98 and 103%, and precision around +/-2%.

Spectrofluorimetric determination of acrivastine in spiked human urine and pharmaceuticals.

A spectrofluorimetric method to determine acrivastine is proposed and applied to its determination in human urine and pharmaceuticals. The fluorimetric method allows the determination of 58-2000 ng ml(-1) of acrivastine in aqueous solutions containing acetic acid-sodium acetate buffer (pH 6.5) with lambda(exc)=230 nm and lambda(em)=380 nm.