Development and Validation of RP-HPLC Method for Simultaneous Estimation of Three-Component Tablet Formulation Containing Acetaminophen, Chlorzoxazone, and Aceclofenac

Abstract

The present work describes a simple reversed-phase high-performance liquid chromatographic method that has been developed and validated for simultaneous estimation of acetaminophen, chlorzoxazone, and aceclofenac in tablet dosage form. The estimation was carried out on an Luna C₁₈ (5 µm × 25 cm × 4.6 mm i.d.) column using a mixture of buffer, methanol, and acetonitrile in the ratio 215:130:155 with final pH of 6.5 as a mobile phase, at a flow rate of 1.5 ml/min. Ultraviolet (UV) detection was performed at 275 nm. Total run time was 10 min; these three drugs (acetaminophen, chlorzoxazone, and aceclofenac) were eluted at the retention times of 2.055, 5.096, and 7.605 min respectively. The method was validated for accuracy, precision, linearity, specificity, and sensitivity as per ICH norms.. From the validation study it was found that the method is specific, rapid, accurate, precise, and reproducible. Calibration curves were linear over the concentration ranges of 5–50 µg/ml for acetaminophen and chlorzoxazone, and 5–30 µg/ml for aceclofenac. All the validation study was found statistically significant because all the statistical parameters were within the acceptance range (i.e., COV % < 2.0 and S.D. < 1.0 for both accuracy and precision). The limit of detection (LOD) values were 16.2, 14.6, and 4.8 ng/ml, and LOQ values were 49.0, 46.5, and 14.5 ng/ml for acetaminophen, chlorzoxazone, and aceclofenac respectively. High recovery and low COV % revealed the reliability of the method for quantitative study of three drugs in Micronac-MR tablets. The method is a rapid and cost-effective quality-control tool for routine quantitative analysis of acetaminophen, chlorzoxazone, and aceclofenac in tablet dosage form.

Keywords:

• Aceclofenac
• acetaminophen
• chlorzoxazone
• ICH guidelines
• RP-HPLC

Acknowledgments

The authors are thankful to Zest Pharma, 275 Sector-F, Sanwer Road, Indore, India, for providing gift samples of paracetamol; Uni Drugs Innovative Pharma Technologies Ltd., Indore, India, for chlorzoxazone; and Aristo Pharma Ltd., Mandideep, India, for aceclofenac. One of the authors is thankful to Suresh Prajapati for technical assistance.

Notes

Notes. ACT: acetaminophen, CHL: chlorzoxazone, ACL: aceclofenac, COV: coefficient of variation.

^a Average of six determinations.

Notes. ACT: acetaminophen, CHL: chlorzoxazone, ACL: aceclofenac, SD: standard deviation, COV: coefficient of variation, SE: standard error.

^a Average of six estimation of tablet formulation.

Notes. Rt: retentiontime, T_f: tailing factor, k′: capacity factor, N: theoretical plate number, Rs: resolution.

Notes. ACT: acetaminophen, CHL: chlorzoxazone, ACL: aceclofenac, COV: coefficient of variation.

^a Average of three estimations at each level of recovery.