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1 2023

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High performance thin layer chromatography (HPTLC) method development and validation for the simultaneous determination of paracetamol, caffeine, chlorpheniramine and phenylepherine in tablet formulation

Arage Almaz, Layloff Thomas, Hymete Ariaya, Ashenef Ayenew

Acta Chromatographica

2023

Vol. 35, no. 2

170--178

A rapid, selective, and precise high performance thin layer chromatographic method was developed and validated for the simultaneous analysis of paracetamol, caffeine, phenylephrine and chlorpheniramine in tablets. The chromatographic analysis was carried out on glass plates pre-coated with silica gel 60 F254 as a stationary phase. The optimized mobile phase was methanol : n-butanol : toluene : acetic acid (8:6:4:0.2 v/v). TLC chamber of 10 × 20 cm was used with saturation time of 15 min. The retardation factor (RF) for chlorpheniramine, phenylephrine, caffeine and paracetamol was found to be 0.15 ± 0.02, 0.29 ± 0.02, 0.50 ± 0.02, 0.68 ± 0.02 respectively. Detection was carried out at 212 nm. Validation study was done following ICH Q2 (R1) guideline. The regression data for the calibration plots showed good linear relationship with R2 = 0.997 over the concentration range of 300–1,500 ng band⁻¹ for caffeine, R2 = 0.996 over the concentration range of 100–500 ng band⁻¹ for phenylephrine, R2 = 0.996 over the concentration range of 200–600 ng band⁻¹ for chlorpheniramine, R2 = 0.998 over the concentration range of 400–2,400 ng band⁻¹ for paracetamol. The method was validated for precision, accuracy and recovery. Minimum detectable amounts were found to be 304.9 ng band⁻¹, 87.88 ng band⁻¹, 117.18 ng band⁻¹ and 143.06 ng band⁻¹ for caffeine, phenylephrine, chlorpheniramine, and paracetamol respectively while the limit of quantification was found to be 923.95 ng band⁻¹, 266.32 ng band⁻¹, 355.11 ng band⁻¹, and 433.53 ng band⁻¹ in the same order. The method was successfully applied to analyze two marketed tablets in a selective and reproducible manner.