Aptamer-affinity column clean-up coupled with ultra high performance liquid chromatography and fluorescence detection for the rapid determination of ochratoxin A in ginger powder.

Aptamers are single-stranded oligonucleotides with high affinity and specificity and are widely used in targets separation and enrichment. Here, an aptamer-affinity column (AAC) was firstly prepared in-house through a covalent immobilization strategy. Then, ochratoxin A (OTA) in ginger powder was absorbed and enriched using the new aptamer-based clean-up technology for the first time, and was further analyzed by ultra high performance liquid chromatography with fluorescence detection. After optimization, the average recoveries for blank samples spiked with OTA at 5, 15, and 45 μg/kg ranged from 85.36 to 96.83%. Furthermore, the AAC exhibited a similar accuracy as an immunoaffinity column to clean up OTA in ginger powder. Above all, it exhibited better reusability, twice that of the immunoaffinity column, had lower toxicity and cost, and took less time. Of 25 contaminated ginger powder samples, OTA contamination levels ranged from 1.51 to 4.31 μg/kg, which were lower than the European Union (EU) regulatory limits. All the positive samples were further confirmed by ultra-fast LC with MS/MS. In conclusion, the method of clean-up based on the AAC coupled to ultra-HPLC with fluorescence detection was rapid, specific, and sensitive for the quantitative analysis of OTA in a complex matrix.

[1]  Jan Bocianowski,et al.  Natural occurrence of fumonisins and ochratoxin A in some herbs and spices commercialized in Poland analyzed by UPLC-MS/MS method. , 2013, Food microbiology.

[2]  E. Peyrin Nucleic acid aptamer molecular recognition principles and application in liquid chromatography and capillary electrophoresis. , 2009, Journal of separation science.

[3]  Chifang Peng,et al.  Fabricated aptamer-based electrochemical "signal-off" sensor of ochratoxin A. , 2010, Biosensors & bioelectronics.

[4]  V. Pichon,et al.  New extraction sorbent based on aptamers for the determination of ochratoxin A in red wine , 2011, Analytical and bioanalytical chemistry.

[5]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[6]  Meihua Yang,et al.  Development and validation of an accurate and rapid LC-ESI-MS/MS method for the simultaneous quantification of aflatoxin B1, B2, G1 and G2 in lotus seeds , 2013 .

[7]  Masayuki Endo,et al.  Chemical Approaches to DNA Nanotechnology , 2009, Chembiochem : a European journal of chemical biology.

[8]  X. Le,et al.  Selection and analytical applications of aptamers , 2006 .

[9]  R. Manderville,et al.  Ochratoxin A: An overview on toxicity and carcinogenicity in animals and humans. , 2007, Molecular nutrition & food research.

[10]  Ioanis Katakis,et al.  Aptamers: molecular tools for analytical applications , 2008, Analytical and bioanalytical chemistry.

[11]  Meihua Yang,et al.  Molecularly imprinted polymer-based solid phase clean-up for analysis of ochratoxin A in ginger and LC-MS/MS confirmation , 2013 .

[12]  J. Frisvad,et al.  Mycotoxin production and postharvest storage rot of ginger (Zingiber officinale) by Penicillium brevicompactum. , 2005, Journal of food protection.

[13]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[14]  A. Pardi,et al.  NMR chemical exchange as a probe for ligand-binding kinetics in a theophylline-binding RNA aptamer. , 2009, Journal of the American Chemical Society.

[15]  F. Accensi,et al.  Occurrence of Aspergillus species in mixed feeds and component raw materials and their ability to produce ochratoxin A , 2004 .

[16]  J. Cruz-Aguado,et al.  Fluorescence polarization based displacement assay for the determination of small molecules with aptamers. , 2008, Analytical chemistry.

[17]  J. M. Soriano,et al.  Rapid determination of ochratoxin A in cereals and cereal products by liquid chromatography. , 2004, Journal of chromatography. A.

[18]  Meihua Yang,et al.  Simultaneous determination of aflatoxin B(1) and ochratoxin A in licorice roots and fritillary bulbs by solid-phase extraction coupled with high-performance liquid chromatography-tandem mass spectrometry. , 2013, Food chemistry.

[19]  Yau Yan Lim,et al.  Effects of different drying methods on the antioxidant properties of leaves and tea of ginger species , 2009 .

[20]  Z. Kučerová,et al.  Application of heptapeptides containing D-amino acid residues immobilized to magnetic particles and Sepharose for the study of binding properties of gastric aspartic proteases. , 2012, Journal of separation science.

[21]  Xiao-He Xiao,et al.  Simultaneous multi-mycotoxin determination in nutmeg by ultrasound-assisted solid-liquid extraction and immunoaffinity column clean-up coupled with liquid chromatography and on-line post-column photochemical derivatization-fluorescence detection. , 2013, The Analyst.

[22]  A. McCarthy,et al.  Ginger (Zingiber officinale) and chemotherapy-induced nausea and vomiting: a systematic literature review. , 2013, Nutrition reviews.

[23]  J. Cruz-Aguado,et al.  Determination of ochratoxin a with a DNA aptamer. , 2008, Journal of agricultural and food chemistry.

[24]  A. De Girolamo,et al.  Analytical performances of a DNA-ligand system using time-resolved fluorescence for the determination of ochratoxin A in wheat , 2012, Analytical and Bioanalytical Chemistry.

[25]  Linda Monaci,et al.  Determination of ochratoxin A in foods: state-of-the-art and analytical challenges , 2004, Analytical and bioanalytical chemistry.

[26]  Y. Shukla,et al.  Cancer preventive properties of ginger: a brief review. , 2007, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[27]  M. DeRosa,et al.  Determination of ochratoxin A in wheat after clean-up through a DNA aptamer-based solid phase extraction column. , 2011, Food chemistry.

[28]  Lin Lu,et al.  Aptamer-targeted magnetic nanospheres as a solid-phase extraction sorbent for determination of ochratoxin A in food samples. , 2011, Journal of chromatography. A.

[29]  F. Fry,et al.  Determination of aflatoxins B1, B2, G1, and G2 and ochratoxin A in ginseng and ginger by multitoxin immunoaffinity column cleanup and liquid chromatographic quantitation: collaborative study. , 2008, Journal of AOAC International.