Development and validation of an HPLC Method for Determination of EDTA in cleaning validation samples

, and reliable High Performance Liquid Chromatography (HPLC) method was developed and validated for the determination of Ethylenediaminetetraacetic acid (EDTA) in cleaning validation samples. The method aims to ensure effective monitoring of residual EDTA on pharmaceutical manufacturing equipment to prevent cross-contamination and maintain product quality. Chromatographic separation was achieved using a Hamilton PRP X-100 (150 × 4.6 mm, 10 µm) column with a mobile phase consisting of sulfuric acid, copper sulfate, methanol, and isopropyl alcohol. The flow rate was maintained at 0.8 mL/min, and detection was carried out at 240 nm with a run time of 8 minutes. The method was validated according to standard regulatory guidelines by evaluating parameters such as system precision, linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, specificity, solution stability, and intermediate precision. The system precision showed excellent reproducibility with a %RSD of 0.8%. The method exhibited strong linearity over a concentration range from LOQ to 300%, with a correlation coefficient of 0.9999. The LOD and LOQ were found to be 0.0121 µg/mL and 0.0242 µg/mL, respectively, indicating high sensitivity. Recovery studies for both swab and plate methods were within acceptable limits (80–120%), with average recoveries around 100%. The method demonstrated good specificity with no interference at the retention time of EDTA and showed stability of solutions up to 72 hours. The validated method is precise, accurate, and robust, making it suitable for routine cleaning validation studies in pharmaceutical industries to ensure compliance with quality standards.