CHROMATOGRAPHIC DETERMINATION OF CHEMICAL COMPONENTS OF

该项目的目标是开发和验证生物制品的化学成分的方法，其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)皮肤试验阳性对照组胺；7)变应原贴片试验中有机、天然产物和复杂合成混合物组分；8)炭疽疫苗中苯并氯铵。目前正在进行的工作包括以下方法开发项目：1)基于USP <341>“抗菌剂-含量”对苯酚和2-苯氧乙醇采用填充柱的现有CBER气相色谱方法进行改造，使之适用于宽口径毛细管柱。宽口径毛细管柱的优点包括通过更高的色谱效率提高分析和增强特异性。由于在许多致敏性萃取物样品中存在的甘油吸附到固定相并导致“鬼峰”和色谱容量因子的变化而引起的潜在问题正在得到解决。2)在CBER的HPLC检测甘油的过程中，正在研究HPLC检测食品中致敏提取物的分析结果比电位滴定法和气相色谱法低。目前正在比较不同技术的分析回收率，并对程序进行修改以提高回收率。3)建立了高效液相色谱法测定炭疽疫苗中抗菌防腐剂苄索氯铵的方法。由于与氢氧化铝佐剂或疫苗基质的明显相互作用，在分析回收该化合物时遇到了困难。分离该化合物的样品制备技术正在研究中。将与产品制造商使用的滴定法获得的结果进行比较。

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 <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 anthrax vaccine. Difficulties have been encountered in the analytical recovery of this compound due to apparent interactions with the aluminum hydroxide adjuvant or the vaccine matrix. Sample preparation techniques to isolate this compound are being investigated. A comparison with results obtained by the titrimetric procedure utilized by the product's manufacturer will be performed.