Evaluation on Removal Effect of Dialyzer to Protein-bound Uremic Toxins Based on High Performance Liquid Chromatography- Triple Quadrupole Mass Spectrometry
  
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KeyWord:protein-bound uremic toxins  dialyzer  simulated fluid  high performance liquid chromatography-triple quadrupole mass spectrometry  removal effect
  
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MU Chang-jun,XU Jie,ZHANG Jie-min,WANG Lei,WANG Yu-feng,JIANG Jun-jie Shan Dong WEGO Blood Purification Products Co. Ltd,Weihai ,China
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Abstract:
      A method for evaluation on the removal effect of dialyzer to protein-bound uremic toxins(PBUTs) was established based on high performance liquid chromatography-triple quadrupole mass spectrometry.The dialysis simulated fluid with the same concentration of PBUTs as the dialysis patients’ blood was prepared instead of blood to conduct dialysis treatment in vitro due to the difficulty to obtain clinical blood samples and the complexity of patient blood composition,and the simulated fluid before and after dialysis was taken as the sample to be tested.After acetonitrile precipitation of protein and ultrafiltration centrifugation,the samples were separated on a Hypersil GOLD VANQUISH C18(100 mm × 2.1 mm,1.9 μm) column by gradient elution,using 5 mmol/L ammonium acetate solution and methanol as mobile phases.The mass spectrometric analysis was carried out under negative ion selective reaction monitoring mode(SRM),and the internal standard method was used for quantitative analysis of 3 PBUTs.The calibration curves of 3 PBUTs showed good linear relationships in the concentration range of 25-1 000 ng/mL,and the correlation coefficients(r) were more than 0.99.The limits of quantitation(LOQs) were 25 ng/mL.The average recoveries for 3 analytes at three added concentrations of 75,250 and 750 ng/mL ranged from 92.5%-114%,with the intra-day and inter-day RSDs both less than 5.0% and the matrix effects of 80.6%-105%.This method had the characteristics of low cost,simple pretreatment,short run-time and high accuracy,which provided a reliable technical support for evaluation on the removal effect of dialyzer to PBUTs.
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