A Study On Protein Adsorption Behavior Using Silver Capillary Column Packed With Hollow Fiber

To obtain the correct data of drugs by using HPLC analysis of biological samples, usually macromolecular compounds like proteins need to be removed from the samples prior to HPLC analysis for they may interfere the HPLC analysis. A novel, simple and miniature pretreatment column made of silver capillary and packed with hollow fiber (polypropylene) has been developed and it was applied to pretreatment of biological samples prior to HPLC analysis. The mechanism of pretreatment is proteins can absorb onto hydrophobic surface or metal surface. When after multiple times pretreatment, column would lost efficiency, through applying electric field inside the pretreatment column, adsorbed proteins would desorption and pre-column would functional again. In this research, we made a pretreatment column which can build electric field in the capillary, in order to remove the proteins adsorbed on the pre-column, recover column absorption ability. We have make a certain this column can eliminate the proteins from mixture sample and absorb proteins into pretreatment column, also investigate how flow rate, electric field type and electric intensity influencing protein adsorption amount, results shows that under0.025mL min-1, similar protein concentrations to urine sample could entirely adsorbed onto pretreatment column. In order to prove pre-column has no interference with drug detection, we using Salvianolic acid B as the template compound to evaluate the feasibility of this pretreatment method. Under the optimized conditions we process the eliminating experiment, relative standard deviation(RSD) is below2.0%, removal rate of protein is99.57%and recovery rate of drug is30.15%. In the end, we have prove that under the square-wave pulse type electric field, electric field strength is3.0V, adsorbed protein is desorption and flushing out from the pre-column, make sure that pre-column can be multiple used. Advantages of this method is it can apply for on-line protein elimination,avoid extra operation step, reducing the loss of target compounds. Moreover, compare with the previous method, pre-column can achieve recycling by applying electric field to eliminate the adsorption protein, make sure that pre-column always possess the adsorption ability. This column provided a simple, fast, effective and friendly to environment method for assaying drugs in the urine. In the future, it has a great potential for pretreatment other biological samples.