SIMULTANEOUS HPLC DETERMINATION AND VALIDATION OF AMPHETAMINE, METHAMPHETAMINE, CAFFEINE, PARACETAMOL AND THEOPHYLLINE IN ILLICIT SEIZED TABLETS

摘要

Objective: The main objective of current study develop and validate HPLC method, rapid, precise, accurate and specific for the separation and determination of paracetamol (PAR), theophylline (THE), amphetamine (AM), methamphetamine (MAM) and caffeine (CAF) in pure form and in illegal formulations. Method: This method is based on HPLC separation of the five drugs on ZORBAX ODS column (250×4.6mm, 5μ). The mobile phase was prepared by mixing buffer (containing 1% of orthophosphoric acid 85% and 1% diethylamine 99%, pH 2.6), acetonitrile and methanol in the ratio 85:10:5 v/v/v. The flow rate is 1.5 ml/min, with isocratic elution and UV detection at 210nm. The retention times of paracetamol, theophylline, amphetamine, methamphetamine and caffeine were found to be at 3.67, 4.27, 5.44 6.72 and 8.40 min, respectively. Results: The linear regression analysis data for calibration plots showed a good linear relationship over a concentration range of 0.39-100 μg/ml for paracetamol, 0.78-100 μg/ml for theophylline, 1.5-100 μg/ml for amphetamine, 1.25-100 μg/ml for methamphetamine and 0.39-100 μg/ml caffeine respectively. The mean values of the correlation coefficient, slope and intercept were 9996, 36.540 and -10.164 for paracetamol, 9999, 37.324 and -19.787for theophylline, 9998, 14.688 and -12.797 for amphetamine, 9995, 13.355 and 14.013 for methamphetamine and 1, 35.229 and 0.048 for caffeine. The method was validated as per the ICH guidelines. The limit of detection (LOD) and limit of quantification (LOQ) was 0.18 and 0.53 μg/ml for paracetamol, 0.21 and 0.63 μg/ml for theophylline, 0.788 and 2.365 μg/ml for amphetamine, 0.717 and 2.15 μg/ml for methamphetamine and 0.22 and 0.66 μg/ml for caffeine. Conclusion: The developed and validated HPLC method and the statistical analysis showed that the method is repeatable and selective for estimation of the five studied drugs