Chromatographic analysis of chemicals in bioproducts

总结：本项目的目标是开发和验证生物制品化学成分的方法，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。 其中包括：1）在多用途胃肠外制剂如变应原提取物和细菌疫苗中用作抗微生物防腐剂的苯酚，2）在变应原提取物中用作防腐剂和/或稳定剂的甘油，3）在灭活脊髓灰质炎病毒疫苗和组合细菌疫苗中用作防腐剂的2-苯氧基乙醇，4）在流感病毒疫苗、乙型肝炎B疫苗和其他产品中用作灭活剂的甲醛，5）人血清白蛋白和葡聚糖体积膨胀剂中的氯化物，6）阳性皮肤试验对照中的组胺，7）过敏原斑贴试验的有机、天然产物和复杂合成混合物组分。 目前正在进行的工作包括以下方法开发项目： 1)根据USP<341>“抗菌剂-含量”对苯酚和2-苯氧基乙醇的现有CBER气相色谱法进行调整，将填充柱用于大口径毛细管柱。 大口径毛细管柱的优点包括通过更高的色谱效率获得更高的分析和增强的特异性。 一个潜在的问题所产生的吸附甘油存在于许多过敏性提取物样品的固定相和所产生的“鬼”峰和色谱容量因子的变化正在解决。 2)在CBER的甘油HPLC程序中，正在研究应用于食品过敏性提取物时，相对于电位滴定和气相色谱法，HPLC观察到的分析结果较低。 沿着对不同技术的分析回收率进行了比较，并对程序进行了修改，以提高回收率。 第三章 抗菌防腐剂苄索氯铵的高效液相色谱分析方法的建立。在过去的一年中，HPLC条件已经优化，并开发了样品制备程序。 通过酸化和过滤完成苄索氯铵从疫苗基质中的解离。该方法已得到部分验证，正在继续研究。

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.