Comparison of Ion-Pairing and Reversed Phase Liquid Chromatography in Determination of Sulfamethoxazole and Trimethoprim

Abstract

Two simple, rapid, and sensitive HPLC methods have been developed for the simultaneous determination of sulfamethoxazole and trimethoprim in their pure and dosage forms, one utilizing reversed phase HPLC and the other ion-pair HPLC. In the reversed phase HPLC method (A) the mobile phase consists of 0.05% aqueous solution of formic acid with pH adjusted to 4.5±0.2 with triethylamine : acetonitrile:tetrahydrofuran 50 : 49 : 1 (v/v), and the mobile phase pumped at flow rate of 1.0 ml min⁻¹. An Appolo LC18 column (5.0 µm), 250 mm length × 4.6 mm diameter, was utilized as the stationary phase. Detection was affected spectrophotometrically at 254 nm. In the ion-pair HPLC method (B) the mobile phase consisted of methanol : buffer 35 : 65 (v/v) with the buffer composed of potassium dihydrogen phosphate 0.3 M and sodium heptan sulfonic acid 5.0 mM. To 500 ml of buffer was added 2.0 ml triethylamine, and then the pH was adjusted to 5.0 with phosphoric acid, and the mobile phase was pumped at a flow rate of 1.2 ml min⁻¹. A Hypersil C₁₈ column (5.0 µm), 150 mm length × 4.6 mm diameter, was utilized as the stationary phase. Detection was affected spectrophotometrically at 254 nm. Linearity ranges for sulfamethoxazole and trimethoprim were 1.0–110 and 1.5–98 µg ml⁻¹, respectively, with method A and 0.5–100 and 1.0–125 µg ml⁻¹, respectively, with method (B). Minimum detection limits obtained were 0.1969 and 0.3451 µg ml⁻¹ for sulfamethoxazole and trimethoprim, respectively, with method A, and 0.1377 and 0.2454 µg ml⁻¹ with method (B). The proposed methods were further applied to the analysis of tablets containing the two drugs, and the results were satisfied.

Keywords:

• Dosage forms
• ion-pair
• reversed phase liquid chromatography
• sulfamethoxazole
• trimethoprim

Notes

Regression equation: A = a + bC, where A is the area under peak and a, is the intercept.

*Average of five determinations.

A: refer to method A, reversed phase HPLC.

B: refer to method B, ion-pair HPLC.

*RSD for five determinations.

Where K: is capacity factor; N: No. of theoretical plates; R: is resolution time; and As: is asymmetry factor.

*Average of five determinations.