I. Bechet

Determination of six water-soluble vitamins in a pharmaceutical formulation by capillary electrophoresis.

A method was developed for the quantitative analysis of six water-soluble vitamins (thiamine, nicotinamide, riboflavine, pyridoxine, ascorbic acid and pantothenic acid) in a pharmaceutical formulation, using free solution capillary zone electrophoresis (CZE) in uncoated fused silica capillaries and UV detection. The influence of different parameters, such as the nature of the buffer anionic component and buffer concentration on the CZE separation of vitamins was investigated using four vitamins of the B group as model compounds. A good compromise between resolution, analysis time and analyte stability was obtained by use of a 50 mM borax buffer of pH 8.5. This CZE method was found to be very useful for the separation of more complex samples, a mixture of ten water-soluble vitamins being completely resolved in about 10 min. However, cyanocobalamine could not be separated from nicotinamide in this CZE system, the two compounds being in uncharged form at the pH used. These two compounds could easily be resolved by micellar electrokinetic chromatography (MEKC), the anionic surfactant dodecylsulfate being added to the running buffer at 25 mM concentration. In the pharmaceutical formulation, some excipients were found to be adsorbed to the capillary surface, giving rise to a progressive decrease of the electroosmotic flow and consequently to a simultaneous increase of analyte migration times. A capillary wash with sodium hydroxide had to be made between successive runs in order to minimize these effects. Good results with respect to linearity, precision and accuracy were obtained in the concentration range studied for the six vitamins, using nicotinic acid as internal standard.

Resolution improvement by use of carboxymethyl-β-cyclodextrin as chiral additive for the enantiomeric separation of basic drugs by capillary electrophoresis

Three beta-cyclodextrin derivatives--carboxymethyl-, dimethyl- and hydroxypropyl-beta-cyclodextrin--were tested as chiral selectors for the enantioseparation of seven basic drugs in free solution capillary electrophoresis, using buffers made of 100 mM phosphoric acid adjusted to pH 3.0 with triethanolamine in fused silica capillaries thermostatted at 15 degrees C. The best results with respect to chiral resolution were obtained with carboxymethyl-beta-cyclodextrin (CMCD): the enantiomers of all compounds examined were completely resolved with this beta-cyclodextrin derivative. The influence of the CMCD concentration on the migration times, the apparent electrophoretic mobility difference and the resolution of the drug enantiomers was investigated thoroughly. Particularly impressive resolution values, up to 23.7, were obtained for several compounds in these capillary electrophoretic systems, using CMCD in the 5-15 mM concentration range.

Separation of nonsteroidal anti-inflammatory drugs by capillary electrophoresis using nonaqueous electrolytes.

The aim of the present work was to investigate the separation of nonsteroidal anti‐inflammatory drugs (NSAIDs: niflumic acid, flufenamic acid, piroxicam, alclofenac, tiaprofenic acid, flurbiprofen, suprofen, ketoprofen, naproxen, indomethacin, carprofen, indoprofen, sulindac) in capillary electrophoresis (CE) using completely nonaqueous systems. The influence of different parameters such as nature and proportion of organic solvent (methanol, acetonitrile, 2‐propanol), apparent pH (ranging from 7 to 9) and temperature (ranging from 25 to 40oC) on selectivity and migration times were studied systematically in an uncoated fused‐silica capillary. A nonaqueous electrolyte made of 50 mM ammonium acetate — 13.75 mM ammonia in methanol proved to resolve 11 NSAIDs at 25oC and 13 NSAIDs at 36oC, both within 13 min and without a modifier besides the methanol itself. The same buffer containing 30% acetonitrile provides a satisfactory separation for 13 NSAIDs within 14 min at 25oC.

Determination of verapamil and norverapamil in human plasma by liquid chromatography: Comparison between a liquid—liquid extraction procedure and an automated liquid—solid extraction method for sample preparation

A conventional liquid-liquid extraction (LLE) procedure with high-performance liquid chromatography (HPLC) has been developed for the determination of verapamil and its main metabolite, norverapamil, in plasma. After addition of the internal standard, plasma samples were basified with phosphate buffer (pH 9.0) and extracted with a mixture of cyclohexane-dichloromethane. After centrifugation, the organic layer was separated and the analytes were extracted back into a 0.1 N sulphuric acid solution containing 2-aminoheptane. An aliquot of this aqueous phase was then injected directly onto the HPLC column. This LLE procedure has been compared with an automated liquid-solid extraction (LSE) method that has been developed in parallel. Good linearity was obtained using both extraction methods. The absolute recoveries for the two analytes were ca 95% with the automated LSE procedure and slightly lower (ca 84%) for the LLE method. The automated method gives better results with respect to detectability and precision, but the LLE procedure is simpler to develop, requires much less expensive equipment, and remains a useful alternative when the number of samples to be analysed is limited.

Determination of meprobamate in pharmaceutical dosage forms also containing carbromal by liquid chromatography and indirect photometric detection

In a pharmaceutical form also containing carbromal, meprobamate could not be quantified selectively by classical methods described in pharmacopoeias due to a significant interference from carbromal. Consequently, reversed-phase HPLC methods have been developed to separate the two active ingredients using indirect photometric detection to visualize and determine meprobamate which has very poor chromophoric properties. Different parameters influencing the sensitivity of the indirect response, such as the nature of the highly absorbing compound added to the mobile phase (the marker) as well as the methanol content and the pH of this phase, have been studied. Two chromatographic systems containing benzoic acid or cinnamic acid as the marker, have been optimized and validated. Good linearity and reproducibility have been obtained with both systems but the cinnamic acid method has the advantage that meprobamate and carbromal can be determined simultaneously at 273 nm.