The objectives of this project are to develop and validate methodologies for chemical constituents 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 and thrombin as a preservative and/or stabilizer, (3) 2-phenoxyethanol used as a preservative in inactivated Poliovirus Vaccine and combined bacterial vaccines, (4) glutaraldehyde used as inactivating or conjugating agent in vaccines, (5) formaldehyde used as an inactivating agent in Influenza virus vaccine, Hepatitis B Vaccine, etc., (6) chloride in Human Serum Albumin and dextran volume expanders, (7) histamine in Histamine, Positive Skin Test Control, (8) residual tetrachloroethylene in pollen allergenic extracts, (9) residual ethyleneimine in a cytokine product and (10) organic, natural product and complex mixture components of allergen patch tests. Preliminary work has been performed on modifying existing CBER gas chromatographic methodologies that utilize packed columns to wide-bore capillary columns. Solid phase extraction techniques for sample preparation have been investigated. An HPLC method for the determination of glutaraldehyde, involving pre-column derivatization with p-nitrobenzyl-hydroxylamine has been developed. Development and validation of a purge and trap-gas chromatographic method for the determination of tetrachloroethylene in allergenic extracts is in progress along with an HPLC method for the determination of residual ethyleneimine.
