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. 2022 Nov 16;27(22):7920.
doi: 10.3390/molecules27227920.

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

Hari Naga Prasada Reddy Chittireddy  1 , J V Shanmukha Kumar  1 , Anuradha Bhimireddy  2 , Mohammed Rafi Shaik  3 , Althaf Hussain Shaik  4 , Abdulrahman Alwarthan  3 , Baji Shaik  5
Affiliations

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

Hari Naga Prasada Reddy Chittireddy et al. Molecules. .

Abstract

Cross contamination of β-lactams is one of the highest risks for patients using pharmaceutical products. Penicillin and some non-penicillin β-lactams may cause potentially life-threatening allergic reactions. The trace detection of β-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 β-lactam antibiotics must be regulated. The analysis was performed on an XBridge C18 column with 100 mm length, 4.6 mm diameter, and 3.5 µm particle size at an oven temperature of about 40 °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 (R2 > 0.99). This sensitive and fast method was fit-for-purpose for detecting and quantifying trace amounts of β-lactam contamination, monitoring cross contamination in facility surface cleaning, and determining the acceptable level of limits for regulatory purposes.

Keywords: beta-lactam ring; ceftiofur; cephalosporin antibiotics; cephapirin; reactor rinse.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Structures of Ceftiofur, Ceftiofur Hydrochloride, Ceftiofur Sodium, Cephapirin, Cephapirin sodium, and Cephapirin Benzathine.
Scheme 1
Scheme 1
Process flow of cleaning and analysis.
Figure 2
Figure 2
MS/MS chromatogram of blank, cephapirin and ceftiofur standard solution.
Figure 3
Figure 3
MS/MS LOD and LOQ chromatogram of cephapirin and ceftiofur standard solutions.
Figure 4
Figure 4
Fragmentation pattern of Cephapirin.
Figure 5
Figure 5
Fragmentation pattern of Ceftiofur.
See this image and copyright information in PMC

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References

    1. Fabio P., Alessandro C., Stefania S., Iolanda G., Elena M., Aldo C. The requirements for manufacturing highly active or sensitising drugs comparing Good Manufacturing Practices. Acta Biomed. Ateneo Parm. 2019;90:288. - PMC - PubMed
    1. Bush K., Bradford P.A. β-Lactams and β-lactamase inhibitors: An overview. Cold Spring Harb. Perspect. Med. 2016;6:a025247. doi: 10.1101/cshperspect.a025247. - DOI - PMC - PubMed
    1. Picard M., Robitaille G., Karam F., Daigle J.-M., Bédard F., Biron É., Tardif M.R., Lacombe-Barrios J., Bégin P. Cross-reactivity to cephalosporins and carbapenems in penicillin-allergic patients: Two systematic reviews and meta-analyses. J. Allergy Clin. Immunol. Pract. 2019;7:2722–2738.e2725. doi: 10.1016/j.jaip.2019.05.038. - DOI - PubMed
    1. Macy E., Contreras R. Health care use and serious infection prevalence associated with penicillin “allergy” in hospitalized patients: A cohort study. J. Allergy Clin. Immunol. 2014;133:790–796. doi: 10.1016/j.jaci.2013.09.021. - DOI - PubMed
    1. Pichichero M.E., Zagursky R. Penicillin and cephalosporin allergy. Ann. Allergy Asthma Immunol. 2014;112:404–412. doi: 10.1016/j.anai.2014.02.005. - DOI - PubMed