Chromatographic analysis of chemicals in bioproducts

摘要：该项目的目标是开发和验证生物制品中化学成分的方法，这些化学成分的浓度对产品的稳定性、有效性或安全性至关重要。这些包括:1)苯酚在致敏提取物和细菌疫苗等多用途静脉注射中用作抗菌防腐剂，2)甘油在致敏提取物中用作防腐剂和/或稳定剂，3)2-苯氧乙醇在灭活脊髓灰质炎病毒疫苗和联合细菌疫苗中用作防腐剂，4)甲醛在流感病毒疫苗、乙型肝炎疫苗和其他产品中用作灭活剂，5)氯化物在人血清白蛋白和葡聚糖体积膨胀剂中用作灭活剂，6)组胺阳性皮肤试验对照，7)有机、天然产物和复杂合成混合物成分的过敏原斑贴试验。

Summary: 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. Current work in progress includes the following methods development projects: 1) The adaptation of existing CBER gas chromatographic methods based on USP <341> "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 the last year, HPLC conditions have been optimized and a sample preparation procedure developed. Dissociation of benzethonium chloride from the vaccine matrix was accomplished by acidification and filtration. The method has been partially validated, continuing studies are in progress.