Degradation Of Aristolochic Acid Ⅰ And Development Of Rapid Immunochromatographic Test Strips For Its Detection

Aristolochic acid I（AA-I）is mainly isolated from Aristolochia plants and is structurally similar to nitrophilic carboxylic acid,which is nephrotoxic and potent carcinogenic,and is mainly found in herbal medicines such as Aristolochia,antennal vine,and viburnum.Therefore,it is important to protect people’s health by degrading the aristolochic acid I in herbal medicine and transforming it into a non-toxic product,as well as detecting aristolochic acid I in herbal medicine.L-cy converts toxic aristolochic acid I into non-toxic aristolochic acid I by reducing the nitro（-NO₂）to hydrogen（-H）on aristolochic acid I.However,this degradation process is subject to many factors.However,this degradation process was affected by many factors,including the molar concentration of L-cy,the volume ratio of L-cy to aristolochic acid I,the reaction p H,and the reaction temperature.In this study,the optimal degradation conditions under the influence of the above conditions were explored and finally applied to the actual samples.Currently,for the detection of aristolochic acid I,the most commonly used detection methods are such as high performance liquid phase,but such methods are not only time consuming but also require professional personnel to operate.Immunochromatographic rapid test strips,which have the advantages of high specificity,rapidity and high efficiency.In this study,we developed immunochromato-graphic rapid test strips to achieve rapid and efficient detection of aristolochic acid I.In order to ensure the validity of the test strips,we developed a method for the detection of aristolochic acid I.In order to ensure the effectiveness and longevity of the test strips,the performance evaluation of the test strips and the comparison of the results with those of high performance liquid chromatography were carried out.Firstly,AA-I was degraded by L-cysteine,and the AA-I standards were quantified by high performance liquid chromatography after 1,2,3,4 and 5 h of degradation,and the degradation effects were investigated under different experimental conditions at different time periods.By comparing the degradation efficiency under different conditions,the best degradation effect was finally established at 0.15 M L-cysteine,volume ratio of 3:1,p H 6 and temperature of 60℃,and this condition was applied to the actual samples.Second,the development of nano-selenium immunochromatographic rapid detection test strips.At present,in the field of immunochromatographic rapid test strips,the most commonly used probe is nanogold,and this study innovated on the probe by introducing selenium nanoparticles.Colloidal selenium is faster,simpler and more economical to synthesize compared to colloidal gold.The colloidal selenium prepared in this study was prepared by reducing selenite（H₂Se O₃）to selenium particles using vitamin C（VC）in the presence of stabilizer.In the preparation of colloidal selenium,the stabilizer selected in this study was polyethylene glycol 8000（PEG8000）sodium dodecyl sulfate（SDS）,and several orange colloidal selenium solutions with characteristics such as uniformly dispersed,spherical shape were successfully prepared by changing PEG8000,SDS,VC and H2Se O3.Later,PEG8000 0.25 g,0.1 m L 0.1 M SDS,2.5 m L 0.32 M VC,2.5 m L 0.04 M H₂Se O₃ were finally optimized to make 50m L of colloidal selenium solution.The nano-selenium was successfully coupled with monoclonal antibodies,and after multiple optimization,nano-selenium immunochromatographic test strips with a sensitivity of 2.5 ng/m L were prepared,and the detection was completed in 10 min.In addition,the performance evaluation of the test strips was carried out,mainly focusing on the specificity and stability.In this study,the rapid detection of small molecule toxins such as aristolochic acid II（AA-II）,T-2 toxin（T-2）and aflatoxin B₁（AFB₁）was carried out using the nano-selenium immunochromatographic strips,and the results showed that no cross-reactivity occurred.In order to investigate the stability of the test strips,experiments were conducted under two conditions:4 weeks at room temperature and 8 weeks at a constant temperature of 25℃,after which the test strips were again subjected to experiments,and the experimental results showed that the test strips could be re-detected after long-term storage,and the sensitivity was consistent before and after storage.Therefore,the test strips prepared in this study have been shown to be specific and stable.Finally,this study compared Se ICA with HPLC for the determination of aristolochic acid I in Chinese herbal medicine.Sixteen herbal samples were tested for the presence of AA-I toxin using the prepared test strips and verified by HPLC.The results showed that the results obtained by both methods converged.It can be shown that the nanoselenium immunochromatographic test strips prepared in this study can be directly applied in the field for the detection and analysis.