Development and Validation for Quantification of Cephapirin and Ceftiofur by Ultraperformance Liquid Chromatography with Triple Quadrupole Mass Spectrometry

Cross contamination of beta-lactams is one of the highest risks for patients using pharmaceutical products. Penicillin and some non-penicillin beta-lactams may cause potentially life-threatening allergic reactions. The trace detection of beta-lactam antibiotics in cleaning rinse solutions of common reactors and manufacturing aids in pharmaceutical facilities is very crucial. Therefore, the common facilities adopt sophisticated cleaning procedures and develop analytical methods to assess traces of these compounds in rinsed solutions. For this, a highly sensitive and reproducible ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the analysis of Cephapirin and Ceftiofur. As per the FDA guidelines described in FDA-2011-D-0104, the contamination of these beta-lactam antibiotics must be regulated. The analysis was performed on an XBridge C-18 column with 100 mm length, 4.6 mm diameter, and 3.5 mu m particle size at an oven temperature of about 40 degrees C. The mobile phase was composed of 0.15% formic acid in water and acetonitrile as mobile phases A and B, and a flow rate was set to 0.6 mL/min. The method was validated for Cephapirin and Ceftiofur. The quantification precision and accuracy were determined to be the lowest limit of detection 0.15 parts per billion (ppb) and the lowest limit of quantification 0.4 ppb. This method was linear in the range of 0.4 to 1.5 ppb with the determination of coefficient (R-2 > 0.99). This sensitive and fast method was fit-for-purpose for detecting and quantifying trace amounts of beta-lactam contamination, monitoring cross contamination in facility surface cleaning, and determining the acceptable level of limits for regulatory purposes.

Automated summary

Cross contamination of beta-lactams in pharmaceutical products is a high risk for patients. A highly sensitive UHPLC-MS/MS method was developed for the detection of trace contamination of Cephapirin and Ceftiofur. This method is important for monitoring cross contamination in facility surfaces and determining acceptable limits for regulatory purposes.