利用高效液相色谱-飞行时间质谱和高效液相色谱-三重四级杆质谱检测南美白对虾的过敏原

Determination of Allergens of Litopenaeus vannamei by HPLC-QTOF and Triple Quadrupole Mass Spectrometer

为检测南美白对虾的过敏原,并寻找表征过敏原的特征肽段,采用测序级胰蛋白酶对南美白对虾的蛋白进行酶解,经高效液相色谱-飞行时间质谱的分离检测和数据库检索,并建立了三重四级杆多反应监测(MRM)快速检测表征过敏原肽段的方法。结果表明,南美白对虾中共检测出7种过敏原,分别为原肌球蛋白、精氨酸激酶、肌球蛋白轻链、血蓝蛋白亚基L1、血蓝蛋白亚基L2、血蓝蛋白亚基L3、肌钙蛋白C1,对应的肽段条数分别为214、16、84、145、225、163和93条。MRM结果显示,该方法灵敏度高、选择性好,为分析南美白对虾过敏原提供了参考。

To detect allergens of Litopenaeus vannamei and find peptides for representation of allergens,high-performance liquid chromatography coupled to quadrupole time of flight mass spectrometry( HPLC-QTOF) with database search was used to separate and detect all sequencing grade modified trypsin-digested peptides.And multiple reaction monitoring( MRM) method on triple quadrupole mass spectrometer was established for rapid detection of the peptides of allergens.The results showed that there were seven kinds of allergens in Litopenaeus vannamei,which were Lit v 1 tropomyosin,arginine kinase,Lit v 3 allergen myosin light chain,hemocyanin subunit L1,hemocyanin subunit L2,hemocyanin subunit L3 and troponin C1,and the numbers of identified peptides of allergens were 214,16,84,145,225,163 and 93 respectively. The MRM result showed that the established method was of high sensitivity and good selectivity.This paper would providea reference for the analysis of allergens of Litopenaeus vannamei.