Abstract

HPTLC PROFILING AND MICROBIAL ASSAY OF FINISHED PRODUCT RESIDUES IN PRODUCTION LINE EQUIPMENT: A TOOL FOR CLEANING VALIDATION

Kotagiri Ravikanth, Anirudh Sharma, Deepak Thakur* and Pushap Lata

ABSTRACT

A validated cleaning process is a tool for pharmaceutical manufacturers to ensure compliance with the current good manufacturing process (cGMP) and GMP + guidelines. A cleaning-verification assay was developed and validated for residues of feed additives and veterinary medicinal finished products in production line equipment viz. fluidized bed dryer (FBD) and tray dryer (TD). The method utilized cleaning water rinse and cotton swab-sampling procedures and high-performance thin layer chromatography (HPTLC) for the separation and detection of the analytes. Further, non-specific methods with effective parameters like pH and microbial assays were also utilized for the analysis of residues from the rinse samples of the production line equipment. Validation of mentioned HPTLC method for the fingerprinting profiling was done. A linear calibration curve with a regression coefficient (R2) of 0.998 was found. Limit of Detection (LOD) and Limit of Quantification (LOQ) were also determined and found to be 0.02 μg spot-1 and 0.06 μg spot-1, respectively. Recovery studies showed 99.0 % recovery at 50 % spiking and 99.08 at 100 % spiking. In the microbial assay, the total plate count was found to be 213 cfu/mL and 198 cfu/mL. Total yeast and mould count were found to be <10 cfu/mL which complies with the set limit and further, the absence of pathogens like Staphylococcus aureus, Escherichia coli, Salmonella Abony, and Pseudomonas aeruginosa was confirmed. From the obtained results, the developed strategy and method for the cleaning validation of manufacturing equipment is acceptable, simple, and practicable for the determination of residues of finished products in rinse samples of production lines.

Keywords: Cleaning validation, HPTLC, Microbial assay, cotton swab, FBD, TD,