Reuse Of Immunoaffinity Column And Its Repeated Use In The Detection Of Ochratoxin A And Aflatoxin In Traditional Chinese Medicine

ObjectiveTraditional Chinese medicinal materials are highly susceptible to contamination of various mycotoxins due to improper storage,etc.The toxicity of mycotoxins is strong and poses a great threat to the health of humans and livestock.The immunoaffinity column has been widely used as a means of purifying toxins because of its high specificity and sensitivity.However,the cost of the immunoaffinity column is relatively high,and it can only be used once.Once the operation is mistaken,it will lead to great waste,and it is not conducive to the development of large-scale toxin detection.Therefore,it is an irresistible trend to study the regeneration and reuse of immunoaffinity columns to reduce the cost of detection and improve the detection efficiency.In this study,the malt,dried ginger and licorice were used as research objects.The Chinese herbal medicines were spiked,purified by immunoaffinity column,and the recovery rate was used as an indicator to investigate the regeneration performance of the Ochratoxin A and the aflatoxin immunoaffinity column,and the relevant conditions which the immunoaffinity column has the most regenerative can be selected.In order to reduce the cost of toxin purification through the experiments.It is conducive to the development of toxin detection in traditional Chinese medicine.It is in line with the concept of saving,high efficiency and environmental protection.MethodsEstablish sensitive and accurate methods for rapid detection of ochratoxin A and aflatoxin in malt,dried ginger and licorice.The high performance liquid chromatograph-y-fluorescence detector(HPLC-FLD)was also established.The effects of different Chinese herbal medicine matrix,sample pretreatment method,pH value of sample extract and other external factors on the regeneration and reuse of immunoaffinity column were investigated by the toxin plus standard recovery rate.Finaly,the relevant conditions for the most repeated use of ochratoxin and aflatoxin immunoaffinity columns were screened out.ResultsIt can be seen from the experimental results that both the ochratoxin and the aflatoxin immunoaffinity column can be regenerated and reused.The number of uses is affected by factors such as the sample matrix,the sample pretreatment method,and the pH of the extract.By regeneration,the ochratoxin immunoaffinity column can be used up to 8 or 3 times for the spiked raw malt or dried ginger sample and the recovery is guaranteed to exceed 70%.When the sample pretreatment with water as a diluent and the pH of the sample solution are adjusted to 7-8,the number of ochratoxin immunoaffinity column regenerations will increase significantly.The number of aflatoxin four-in-one immunoaffinity column regeneration will vary according to the specific toxin.The aflatoxin immunoaffinity column has the best regeneration when using the sample pretreatment method of Huaan Magnech Bio-Tech Co.The immunoaffinity column can be used 9 times for the licorice and raw malt as matrix and the recovery of aflatoxin G2 and B2 is maintained at an acceptable level.In addition,for immunoaffinity columns,PBS can be used as a good filling solution for column regeneration and reuse.ConclusionsThrough the spiked recovery test of toxins,various relevant conditions for the best regeneration of immunoaffinity columns were successfuly screened,and it was proved that the ochratoxin and aflatoxin immunoaffinity columns can be regenerated and reused,which can reduce the experiment cost.It will provide the ideas and references for large-scale toxin purification experiments.