Qin Lu, Zhang Lei, Jiang Jia-Yi, Wang Chang-Jian, Dou Xiao-Wen, Wan Li, Yang Mei-Hua
College of Pharmacy,Chengdu University of Traditional Chinese Medicine Chengdu 611137,China Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100193,China.
Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100193,China.
Zhongguo Zhong Yao Za Zhi. 2019 Dec;44(23):5072-5077. doi: 10.19540/j.cnki.cjcmm.20190916.201.
An indirect competitive enzyme-linked immunosorbent assay( ic-ELISA) was developed for the rapid detection of ochratoxin A( OTA) in nutmeg( Myristicae Semen),ginger( Zingiberis Rhizoma) and turmeric( Curcumae Longae Rhizoma). The matrix matching standard curve was used instead of the standard curve of sample diluent,and the sample extract and sample diluent were optimized. The sensitivity( IC_(50)) of this method for OTA in nutmeg,ginger and turmeric were determined as 0. 146,0. 157 and 0. 153 ng·m L(-1),respectively and the limits of detection( LODs) were 0. 040,0. 032 and 0. 031 ng·m L(-1),respectively. The recovery of samples ranged from 75. 99% to 122. 3%,with RSD<10%. Two positive samples for nutmeg and one positive sample for turmeric occurred in 50 samples,and the highest OTA contamination value was 1 167. 8 μg·kg~(-1). The results were further confirmed by LC-MS/MS. It shows that the developed ic-ELISA method is simple,rapid and sensitive,and can be applied for rapid and high-throughput screening of OTA in nutmeg,ginger and turmeric,as well as some other CHMs.
建立了一种间接竞争酶联免疫吸附测定法（ic-ELISA），用于快速检测肉豆蔻、生姜和姜黄中的赭曲霉毒素A（OTA）。采用基质匹配标准曲线代替样品稀释剂的标准曲线，并对样品提取物和样品稀释剂进行了优化。该方法对肉豆蔻、生姜和姜黄中OTA的灵敏度（IC50）分别为0.146、0.157和0.153 ng·mL-1，检测限（LOD）分别为0.040、0.032和0.031 ng·mL-(-1)。样品回收率在75.99%至122.3%之间，相对标准偏差（RSD）<10%。50份样品中，肉豆蔻有2份阳性样品，姜黄有1份阳性样品，OTA最高污染值为1167.8 μg·kg-1。结果通过液相色谱-串联质谱（LC-MS/MS）进一步确认。结果表明，所建立的ic-ELISA方法简单、快速、灵敏，可用于肉豆蔻、生姜、姜黄以及其他一些中药材中OTA的快速高通量筛查。
