High-throughput screening of ochratoxin A in Chinese herbal medicines using enzyme-linked immunoassay

QIN Lu1,2 ZHANG Lei2 JIANG Jia-yi2 WANG Chang-jian2 DOU Xiao-wen2 WAN Li1 YANG Mei-hua2

(1.College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, China 611137)
(2.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, China 100193)

【Abstract】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 (IC50) of this method for OTA in nutmeg, ginger and turmeric were determined as 0.146, 0.157 and 0.153 ng·mL−1, respectively and the limits of detection (LODs) were 0.040, 0.032 and 0.031 ng·mL−1, respectively. The recoveries 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 Chinese herbal medicines.

【Keywords】 indirect competitive enzyme-linked immunosorbent assay; Myristicae Semen; Zingiberis Rhizoma; Curcumae Longae Rhizoma; ochratoxin A; rapid screening;


【Funds】 National Natural Science Foundation of China (81872999, 81703699) Medical and Health Science and Technology Innovation Project of the Chinese Academy of Medical Sciences (2017-I2M-1-013) National Key Research and Development Program of China (2016YFE0112900) National Overseas Experts Program (G20190001631)

Download this article


    [1] Lalini R, Kanti B. Ochratoxins-food contaminants: impact on human health [J]. Toxins, 2010, 2 (4): 771.

    [2] Gao X, Li M, Zhang L S. 赭曲霉毒素A的毒性研究进展 [J]. Foreign Medical Sciences (Section of Hygiene), 2005, 32 (1): 51 (in Chinese).

    [3] IARC working group on the evaluation of carcinogenic risks to humans. Some naturally occurring substances: food items and constituents, heterocyclic aromatic amines and mycotoxins [J]. Carcinógenos, 1993, 56: 245.

    [4] European Union. Setting maximum levels for certain contaminants in food stuffs. Commission Regulation (EC) No 1881/2006 [J]. J Eur Uion, 2006, 364: 5.

    [5] Elena C L, Katrien B, Hamide S, et al. Determination of ochratoxin A in black and white pepper, nutmeg, spice mix, cocoa, and drinking chocolate by high-performance liquid chromatography coupled with fluorescence detection: collaborative study [J]. J AOAC Int, 2017, 100 (5): 1458.

    [6] Caballero-Casero N, GarcÍA-Fonseca S, Rubio S. Restricted access supramolecular solvents for the simultaneous extraction and cleanup of ochratoxin A in spices subjected to EU regulation [J]. Food Control, 2018, 88: 33.

    [7] Thirumala-Devi K, Mayo M A, Reddy G, et al. Occurrence of ochratoxin A in black pepper, coriander, ginger and turmeric in India [J]. Food Addit Contam, 2001, 18 (9): 830.

    [8] Yang L, Wang L N, Pan J Y, et al. Determination of ochratoxin A in traditional Chinese medicinal plants by HPLC-FLD [J]. Food Addit Contam, 2010, 27 (7): 989.

    [9] Li H. Determination of Ochratoxin A Content in Chinese Medicinal Materials by Immunoaffinity Column Chromatography Purification-HPLC [J]. China Pharmaceuticals, 2015 (12): 62 (in Chinese).

    [10] Zheng R S, Xiao Q Y, Qiu W, et al. Determination of contaminant ochratoxin A in 10 traditional Chinese medicines and 3 food samples by LC-MS/MS [J]. Chinese Journal of Pharmaceutical Analysis, 2015 (2): 289 (in Chinese).

    [11] Zheng R, Jian L H, Mao D, et al. Determination of ochratoxin A in traditional Chinese medicine [J]. Chinese Traditional Patent Medicine, 2011, 33 (10): 1757 (in Chinese).

    [12] Liu Q F. Application of ELISA for detection of ochratoxin from grain and fodder [J]. China Tropical Medicine, 2005, 5 (5): 1078 (in Chinese).

    [13] Li H J, Li X B, Ding Y B, et al. Rapid detection of ochratoxin A in feed by enzyme-linked immunosorbent assay [J]. Journal of Food Safety & Quality, 2018, 9 (9): 233 (in Chinese).

    [14] Chu X F, Dou X W, Kong W J, et al. Contamination level of aflatoxin B1 in lotus seeds rapid screening by indirect competitive ELISA method [J]. China Journal of Chinese Materia Medica, 2015, 40 (4): 704 (in Chinese).

    [15] Wang S. 易霉变中药材的储藏规范研究——以麦芽、莲子、肉豆蔻为例 [D]. Beijing: Chinese Academy of Medical Sciences & Peking Union Medical College, 2016 (in Chinese).

    [16] Shi J, Huang B, Sun W R, et al. Research on Determination of Ochratoxin A by Improved ELISA Method [J]. Food Science, 2007, 28 (8): 425 (in Chinese).

    [17] Zhang N, Li M, Li P W, et al. Effect of sample matrix on determination of aflatoxin B1 by enzyme-linked immunosorbent assay [J]. Chinese Journal of Oil Crop Sciences, 2014, 36 (3): 404 (in Chinese).

    [18] Wei F, Liao X F, Liu X F, et al. Research progress of pretreatment technology for mycotoxin detection in Chinese Materia medica and complex matrices [J]. China Journal of Chinese Materia Medica, 2018, 43 (17): 3431 (in Chinese).

This Article



Vol 44, No. 23, Pages 5072-5077

December 2019


Article Outline


  • 1 Materials
  • 2 Methods
  • 3 Results and analysis
  • 4 Discussion and conclusion
  • References