Determination of isoniazid among pharmaceutical samples and the patients’ saliva samples by using potassium ferricyanide as spectroscopic probe reagent

A novel method to determine isoniazid with high sensitivity and good selectivity has been established by using potassium ferricyanide as spectroscopic probe reagent. In the presence of potassium ferricyanide, it has been demonstrated that iron(III) is reduced to iron(II) by isoniazid at pH 4.0. In addition, the in situ formed iron(II) reacts with potassium ferricyanide to give soluble prussian blue. The absorbance of soluble prussian blue is measured at the absorption maximum of 735 nm, and the amount of isoniazid can be calculated based on this absorbance. A good linear relationship of the concentration of isoniazid versus absorbance is observed with a linear range of 0.040-8.00 μg mL⁻¹. The linear regression equation is A = −0.00286 + 0.28588C (μg mL⁻¹) with a correlation coefficient of 0.9992. The detection limit (3σ/k) is 0.037 μg mL⁻¹, and its relative standard deviation (R.S.D.) is 0.35% (n = 11). Moreover, the apparent molar absorption coefficient of indirect determination of isoniazid is 3.92 × 10⁴ L mol⁻¹ cm⁻¹. The parameters with regard to determination are optimized, and the reaction mechanism is discussed. This method has been successfully applied to the determination of isoniazid in pharmaceutical samples and saliva samples of patients.