DEVELOPMENT OF A RAPID ASSAY FOR VITAMIN D IN MILK

牛奶中维生素 D 快速测定方法的开发

• 批准号: 2536608
• 负责人: TAI CHENG CHEN
• 金额: $ 10万
• 依托单位: A AND D BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2536608
• 关键词: baby food; binding proteins; bioassay; bioluminescence; biotin; diagnosis design /evaluation; diagnosis quality /standard; enzyme linked immunosorbent assay; food quality /standard; method development; milk; nutrient requirement; nutrition related tag; vitamin D

DESCRIPTION: Vitamin D plays an essential role for the growth and maintenance of healthy bones. Fortification of fresh milk and infant formula with vitamin D has played an important role in eradicating rickets in children, whereas, overfortification can cause vitamin D intoxication and other serious consequences. Thus, it is imperative that milk samples and infant formula have the correct vitamin D concentration as stated on the label. However, the current methods for the determination of vitamin D are very time consuming. It requires highly skilled personnel to perform the experiments and takes several days for completion. By the time they receive the reports, their milk products have already been shipped to the market and purchased. Therefore, the goal of this research project is to develop a rapid, sensitive, and specific assay for vitamin D in milk and infant formula utilizing high affinity binding between biotin and avidin coupled to horseradish peroxidase to develop an enzyme-linked immunosorbent assay (ELISA) for vitamin D. Because the sensitivity of ELISA depends on the detection limit for the calorimetric substrates used, the investigator proposes to utilize chemiluminescence ELISA substrates. The investigator proposes that the sensitivity can be increase at least 3 orders of magnitude over conventional calorimetric assay. The specific aims for this Phase I proposal are to:1. develop a rapid and sensitive non-radioactive assay to determine vitamin D concentration in milk using a combination of chemiluminescence and biotin-avidin technology, and 2. develop a rapid and sensitive non-radioactive assay to determine vitamin D concentration in infant formula using a combination of chemiluminescence and biotin-avidin technology. Eventually, a simple commercial assay kit will be developed for marketing that will allow an on-site analysis of vitamin D contents in milk and infant formula. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述：维生素D对生长和发育起着至关重要的作用。 保持骨骼健康。 强化鲜奶和婴儿 含有维生素D的配方在根除 儿童佝偻病，而，过度强化可导致维生素D 中毒和其他严重后果。 因此， 牛奶样本和婴儿配方奶粉中的维生素D 浓度如标签所示。 然而，目前用于 维生素D的测定非常耗时。 它需要 高技能的人员进行实验，并采取几个 天完成。 当他们收到报告时，他们的牛奶 产品已经被运往市场并被购买。 因此，本研究项目的目标是开发一种快速， 牛奶和婴儿配方奶粉中维生素D的灵敏和特异性测定 利用生物素和抗生物素蛋白之间的高亲和力结合， 辣根过氧化物酶建立酶联免疫吸附测定法 （ELISA）测定维生素D。因为ELISA的灵敏度取决于 所用量热底物的检测限，研究者 提出利用化学发光ELISA底物。 的 研究者建议灵敏度至少增加3 数量级超过常规量热分析。 具体 第一阶段的目标是：1。 开发一种快速灵敏的 非放射性测定法测定牛奶中的维生素D浓度， 化学发光和生物素-亲和素技术的组合，以及2. 开发一种快速、灵敏的非放射性测定方法， 使用以下组合测定婴儿配方食品中的D浓度 化学发光和生物素-亲和素技术。 最终，一个简单的 将开发用于市场的商业测定试剂盒， 现场分析牛奶和婴儿配方奶粉中的维生素D含量。 拟议商业应用：不可用