Citation: XIAO Chang-Bin, LIU Qiu-Tao, DOU Xiao-Wen, YANG Mei-Hua, KONG Wei-Jun, WAN Li. Rapid Detection of Ochratoxin A in Malt by Cytometric Bead Array Based on Indirect Competition Principle[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(4): 625-632. doi: 10.11895/j.issn.0253-3820.151007 shu

Rapid Detection of Ochratoxin A in Malt by Cytometric Bead Array Based on Indirect Competition Principle

XIAO Chang-Bin ^1,2 ,
LIU Qiu-Tao ^2,3 ,
DOU Xiao-Wen ² ,
YANG Mei-Hua ² ,
KONG Wei-Jun ^2,, ,
WAN Li ^1,,

1.
¹ Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China;
2.
² Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China;
3.
³ Pharmacy College, Liaoning Medical University, Jinzhou 121001, China

Corresponding author: KONG Wei-Jun, WAN Li,
Received Date: 23 December 2015
Available Online: 7 March 2016
Fund Project: 本文系国家自然科学基金(Nos.81473346,81274072) (Nos.81473346,81274072)

A cytometric bead array(CBA) method based on indirect competition mode was developed for the sensitive and rapid detection of ochratoxin A(OTA) in malt. The malt samples were extracted by 60% methanol/PBS and the extracts were diluted five times by 20% methanol/PBS, then the supernatant was collected to prepare sample solution for analysis. The fluorescence microsphere surface was labeled with bovine serum albumin-OTA(BSA-OTA) to compete with OTA in the sample for anti-OTA monoclonal antibody(mAb). Then FITC-IgG was added to bind with the captured mAb on the microsphere. Finally, the mean fluorescence intensity of FITC on the surface of microsphere was detected by a BD FACSCalibur analyzer for accurate qualitative and quantitative analysis of OTA. The results showed that the half inhibitory concentration(IC₅₀) was 1.20 ng/mL with a correlation coefficient(R²) of 0.9892, and the limit of detection(LOD) for OTA was 0.12 ng/mL. The average recoveries in malt samples were 93.9%-97.4% with the relative standard deviations(RSDs) less than 3.6% at three spiked levels. Sixteen malt samples were analyzed and OTA was found in two samples with the contents less than 3.83μg/kg, which was lower than the maximum permitted residue level(5.0μg/kg) proposed by the European Union. All the positive samples were confirmed by LC-MS/MS. In this study, the CBA technique based on indirect competition was developed for the first time for successful detection of OTA in malt samples. This method was easy, rapid, sensitive and reliable, and exhibited high potential in the qualitative and quantitative determination of multiple mycotoxins in other complex matrices.
- Cytometric bead array,
- Indirect competition,
- Malt,
- Ochratoxin A,
- Rapid detection
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