Development and validation of an HPLC method for simultaneous determination of trimethoprim and sulfamethoxazole in human plasma.

The combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is used in the treatment of many common infections such as urinary, respiratory and gastrointestinal tract infections. The aim of this study was to determine TMP and SMX simultaneously in human plasma samples by high performance liquid chromatography (HPLC) using antipyrine as the internal standard. Separation of the compounds was achieved on a reverse-phase C8 column packed with 5 microm dimethyl octadecylsilyl bonded amorphous silica (4.6 mm x 250 mm) column using a mobile phase consisted of potassium hydrogen phosphate, acetonitrile, methanol and water adjusted to pH 6.2. The mobile phase was delivered at a flow rate of 1 mL min- and the effluent was monitored using Max plot technique at 25 derees C. Retention times were 5 min for TMP, 7 min for antipyrine and 9 min for SMX. Quantitation limits were 10 ng mL(-1) for TMP and 50 ng mL(-1) for SMX. Our findings indicated that the developed HPLC method was precise, accurate, specific and sensitive for simultaneous determination of TMP and SMX. Proposed HPLC method was successfully applied for the analysis of TMP and SMX in human plasma after oral administration of a co-trimoxazole tablet to human volunteers.

Simultaneous determination of metronidazole and miconazole in pharmaceutical dosage forms by RP-HPLC.

A reversed-phase high performance liquid chromatography (RP-HPLC) method with UV detection is described for the simultaneous determination of metronidazole and miconazole in pharmaceutical dosage forms. Chromatography was carried out on a C18 reversed-phase column, using a mixture of methanol-water (40+60, v/v) as a mobile phase, at a flow rate of 1.0 ml min(-1). Sulfamethoxazole was used as an internal standard and detection was performed using a diode array detector at 254 nm. The method produced linear responses in the concentration ranges 10-70 and 1-20 microg ml(-1) with detection limits 0.33 and 0.27 microg ml(-1) for metronidazole and micanozole, respectively. This procedure was found to be convenient and reproducible for analysis of these drugs in ovule dosage forms.

Determination of theophylline and ephedrine HCL in tablets by ratio-spectra derivative spectrophotometry and LC.

Two methods are described for the determination of theophylline (THP) and ephedrine hydrochloride (EPH) in combined pharmaceutical tablet forms. The first method depends on the use of the first derivative of the ratio-spectra obtained by dividing the absorption spectrum of binary mixtures by a standard spectrum of one of the compounds. The first derivative amplitudes at 231.8 and 250.3 nm were selected for the assay of THP and EPH, respectively. Calibration graphs were established for 20-180 microg ml(-1) for THP and 10-50 microg ml(-1) for EPH. The second method is based on high-performance liquid chromatography on a reversed-phase column using a mobile phase of methanol-water (40+60,v/v) (pH 3) with detection at 217 nm. Linearity was obtained in the concentration range of 5-150 microg ml(-1) for THP and 15-75 microg ml(-1) for EPH. The detection limits for THP and EPH were 0.73 and 0.92 microg ml(-1) by ratio-spectra derivative spectrophotometry and 0.59 and 0.86 microg ml(-1) by HPLC, respectively. The proposed methods were successfully applied to the determination of these drugs in laboratory-prepared mixtures and in tablets. The relative standard deviations were found to be less than 1.5%, indicating reasonable repeatibility of both methods.