Wang Hsiaoling, Del Grosso Alfred V, May Joan C
Laboratory of Analytical Chemistry, Office of Vaccine Research and Review, Center of Biological Evaluation and Research, Food and Drug Administration, 1401 Rockville Pike HFM-406, Rockville, MD 20852, USA.
Biologicals. 2006 Dec;34(4):257-63. doi: 10.1016/j.biologicals.2005.11.004. Epub 2006 Feb 21.
A novel and sensitive HPLC method for the determination of benzethonium chloride (BZC) in anthrax vaccine was developed. Adjuvant Alhydrogel was removed by syringe filter after a simple sample pretreatment - acidification prior to injection. Chromatography was performed by isocratic reverse phase separation with methanol/262 mM ammonium acetate (80/20, v/v) on an endcapped C18 column with diode array detector (DAD). The method showed excellent recovery (100+/-1.5%). The results indicated that this method could accurately determine BZC at the limit of detection (LOD) of 0.5 ppm and the limit of quantitation (LOQ) of 1.5 ppm with dynamic range up to 100 ppm. The comparison of analysis between new HPLC and old titrimetric methods is also reported. The HPLC method is proven to be more accurate and precise with much less vaccine sample and human labor required.
