摘要
小分子半抗原直接包被ELISA方法被用于保健食品中酚酞的检测。将氨基化酚酞交联在6.5%戊二醛处理后的酶标板上,通过间接竞争酶联免疫吸附试验(ic-ELISA)建立酚酞含量检测标准曲线。对其主要影响因素如包被浓度、封闭条件、抗体稀释度等进行优化,并对该方法进行考察。结果表明,方阵滴定试验得到抗体血清的最佳稀释倍数为1︰100,包被半抗原的最佳浓度为4μg/m L。间接竞争ELISA的标准曲线为y=38.795x-22.554, R^2=0.980 2,IC50=74.15 ng/mL,检测范围为12.50~439.93 ng/mL。在检测范围内精密度RSD为4.5%~9.3%。平均加标回收率为89.76%(RSD=7.96%)。试验建立氨基化酚酞直接包被ELISA检测酚酞含量的方法,该方法减少抗体的非特异性结合,具有简便、稳定、灵敏度高等特点。
The ELISA method was applied to the detection of phenolphthalein in diet pills by small molecule hapten direct coating ELISA. The aminophenol phenolphthalein was cross-linked on the ELISA plate after 6.5% glutaraldehyde treatment, and the standard curve of phenolphthalein was established by indirect competitive enzyme-linked immunosorbent assay(ic-ELISA). The main influencing factors such as coating concentration, blocking conditions, and antibody dilution were optimized, and the method was investigated. The result showed that the optimal dilution ratio of antibody serum obtained by square matrix titration was 1︰100, and the optimal concentration of coating hapten was 4 μg/mL. The standard curve for the indirect competition ELISA was y=38.795 x-22.554, R^2=0.980 2, IC50=74.15 ng/mL. The detection range was 12.50-439.93 ng/m L. The relative standard deviation(RSD) was 4.5%-9.3% within the detection range, with the average recovery rate of 89.76%(RSD=7.96%). A method for the determination of phenolphthalein by ELISA was established. The method reduced the non-specific binding of the polyclonal antibody, and had the characteristics of simplicity, stability, and high sensitivity.
作者
杨文斌
李晓娜
管淑玉
YANG Wenbin;LI Xiaona;GUAN Shuyu(College of Chinese Medicine,Guangdong Pharmaceutical University,Guanzhou 511400;Key Laboratory of Digital Quality Evaluation of Chinese Materia Medica of State Administration of TCM,P.R.China,Guanzhou 511400)
出处
《食品工业》
CAS
北大核心
2019年第12期286-289,共4页
The Food Industry
基金
广州市科技计划产学研协同创新重大专项（201508020016）
广东药科大学创新强校工程项目
