The objectives of this project are to develop and validate methodologies for chemical components of biological products whose concentration is vital to product stability, efficacy or safety. These include: 1) phenol used as an antimicrobial preservative in multi-use parenterals such as allergenic extracts and bacterial vaccines, 2) glycerin used in allergenic extracts as a preservative and/or stabilizer, 3) 2-phenoxyethanol used as a preservative in inactivated poliovirus vaccine and combined bacterial vaccines, 4) formaldehyde used as an inactivating agent in influenza virus vaccine, hepatitis B vaccine and other products, 5) chloride in human serum albumin and dextran volume expanders, 6) histamine in positive skin test control, 7) organic, natural product and complex synthetic mixture components of allergen patch tests, 8) benzethonium chloride in anthrax vaccine. Current work in progress includes the following methods development projects: 1) The adaptation of existing CBER gas chromatographic methods based on USP """"""""Antimicrobial Agents - Content"""""""" for phenol and 2-phenoxyethanol that utilize packed columns to wide-bore capillary columns. Advantages of wide-bore capillary columns include greater analytical and enchanced specificity through higher chromatographic efficiency. A potential problem arising from the adsorption of glycerin present in many allergenic extract samples to the stationary phase and resulting """"""""ghost"""""""" peaks and changes in chromatographic capacity factors is being addressed. 2) In the CBER's HPLC procedure for glycerin, lower analytical results observed with HPLC relative to potentiometric titration and gas chromatography when applied to allergenic extracts of foods are being investigated. Analytical recoveries by the different techniques are being compared along with procedural modifications to enhance recovery. 3) The development of an HPLC procedure for the anti-microbial preservative benzethonium chloride in anthrax vaccine. In the last year, an HPLC method has been developed and partially validated. A manuscript has been drafted.
