Quantification of residual EDU ( N-ethyl- N′-(dimethylaminopropyl) carbodiimide (EDC) hydrolyzed urea derivative) and other residual by LC–MS/MS

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 813; no. 1; pp. 103 - 112
• Main Authors: Lei, Q. Paula, Lamb, David H., Shannon, Anthony G., Cai, Xinxing, Heller, Ronald K., Huang, Michael, Zablackis, Earl, Ryall, Robert, Cash, Patricia
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 25.12.2004; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Assay transfer; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; Ethyldimethylaminopropyl Carbodiimide - analogs & derivatives; Ethyldimethylaminopropyl Carbodiimide - analysis; Fundamental and applied biological sciences. Psychology; General pharmacology; Hydrolysis; LC–MS; Mass Spectrometry - methods; Medical sciences; Pharmacology. Drug treatments; Polysaccharide protein conjugate vaccine; Quantification; Reproducibility of Results; Residual material; Sensitivity and Specificity; Validation; Validation; Polysaccharide protein conjugate vaccine; Residual material; Quantification; Assay transfer; LC–MS; Investigation method; Urea
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2004.09.015

An LC–MS/MS method for determination of the break down product of N-ethyl- N′-(3-dimethylaminopropyl) carbodiimide (EDC) urea derivative, EDU, has been developed and validated for monitoring the residual coupling reagents. Results indicate that the method exhibits suitable specificity, sensitivity, precision, linearity and accuracy for quantification of residual EDU in the presence of meningococcal polysaccharide-diphtheria toxoid conjugate vaccine and other vaccine matrix compounds. The assay has been validated for a detection range of 10–100 ng/mL and then successfully transferred to quality control (QC) lab. This same method has also been applied to the determination of residual diaminohexane (DAH) in the presence of EDU. LC–MS/MS has proven to be useful as a quick and sensitive approach for simultaneous determination of multiple residual compounds in glycoconjugate vaccine samples.