The Application Of Immune Affinity Column On Simultaneous Detection Of Three Kinds Of Lean Meat Essence

In this paper，we researched the method to prepare IAC which can detect ractopamine,salbutamol, clenbuterol simultaneously. At the same time，we optimized its elutionconditions and tested the column performance. Finally，we determined the optimum useconditions. At last, we compared this method to method Enzyme-linked immunosorbent，method which was purified by solid phase extraction column and detected by highperformance liquid chromatography，method which was Purified by solid phase extractioncolumn and detected by ultra-performance liquid chromatography-electrospray ionizationtandem mass spectrometry respectively．First， the anti-ractopamine monoclonal aitibody and anti-clenbuterol monoclonalaitibody were both coupled with sepharose4B which was activated by CNBr. Then we madethe immunoaffinity column，optimized the elution condition of the column and tested thecolumn performance. The results showed: The column capacity of ractopamine, salbutamol,clenbuterol were49ng/mL glue，27ng/mL glue，24ng/mL glue respectively. The storage timewas five month. Methanol of3ml was the best eluent. The optimum PH was7-8. The columncould also tolerate aqueous methanol which has methanol less than25%and aqueousacetonitrile which has acetonitrile less than20%.Then，we studied four methods which can detect three kinds of lean meat essencesimultaneously: method Enzyme-linked immune(ELISA), the method was suit for detectionof mass sample.It consumed short time. The method detection limit(LOD) was10ng/g;method which was purified by solid phase extraction column and detected by highperformance liquid chromatography, the average recoveries ranged from72.96%-112.48%.The RSD%was lower than10%. However，this method was not that accurate; method whichwas purified by solid phase extraction column and detected by ultra-performance liquidchromatography-electrospray ionization tandem mass spectrometry. We studied the differencebetween PCX column and MCX column in the recoveries of three materials. The resultsshowed that the two column had the same recovery.The average recovery of the three leanmeat essence ranged from54.88%to101.4%. The RSD%were all lower than3%．The LODof this method was1.3-2ng/g，that was quite accurate．However, this method was soexpensive that some institution could not afford to it; method which was purified by solidphase extraction column and detected by ultra-performance liquidchromatography-electrospray ionization tandem mass spectrometry: The average recovery ofthe three lean meat essence ranged from78.90%to104.00%.The RSD%was lower than2%.The LOD was0.1-1ng/g．From the results，we can see that compared to other methods，the method usingimmunoaffinity column to purify samples has good reproducibility，high recovery and is moresensitive. But，it has flaws at the same time, such as high cost, short storage time and must becryopreserved. It deserves further research．...