A NEW VALIDATED METHOD FOR THE SIMULTANEOUS DETERMINATION OF A SERIES OF EIGHT BARBITURATES BY RP-HPLC

Abstract

A new reversed-phase high performance liquid chromatographic (RP-HPLC) method is developed and validated for the simultaneous determination of barbitone, allobarbitone, phenobarbitone, cyclobarbitone, hexobarbitone, pentobarbitone, secobarbitone and methohexitone compounds in a single analytical run. The method uses a Phenosphere C₁₈ (150 mm × 4.6 mm; 5 μm) column and isocratic elution. The mobile phase consisted of a mixture of methanol-water (50:50, v/v), pumped at a flow rate of 1.0 mL/min. The UV detection is set at 254 nm. The method is validated with respect to accuracy, precision (repeatability and intermediate precision), specificity, linearity, range robustness and stability of analytical solutions. All the parameters examined met the current recommendations for bioanalytical method validation. The method is specific, simple, selective and reliable for routine use in quality control analysis of barbiturates raw materials for final product release.

Keywords:

• allobarbitone
• barbitone
• cyclobarbitone
• hexobarbitone
• method development
• method validation
• methohexitone
• pentobarbitone
• phenobarbitone
• reversed-phase liquid chromatography
• secobarbitone

Notes

*Height equivalent to a theoretical plate.

^a Target concentration corresponding to 100%.

^a The coefficient of variation.