超高效液相色譜-串聯質譜法同時測定魚制品中殘留的7種性激素

張愛芝, 王全林＊, 沈 堅, 張書芬, 陳立仁

(1.寧波市產品質量監督檢驗研究院,浙江寧波315041;2.中國科學院蘭州化學物理研究所,甘肅蘭州730000)

超高效液相色譜-串聯質譜法同時測定魚制品中殘留的7種性激素

張愛芝1, 王全林1＊, 沈 堅1, 張書芬1, 陳立仁2

(1.寧波市產品質量監督檢驗研究院,浙江寧波315041;2.中國科學院蘭州化學物理研究所,甘肅蘭州730000)

以電噴霧離子源(ESI)為電離源,在正離子采集模式下建立了魚制品中7種性激素(甲基炔諾酮、甲基睪酮、丙酸睪酮、醋酸甲羥孕酮、醋酸甲地孕酮、醋酸氯地孕酮、諾龍)的超高效液相色譜-質譜/質譜(UPLC-MS/MS)檢測方法。樣品被酶解后用甲醇提取,提取液經氯化鋅(ZnCl2)去脂、LC-C18和LC-NH2固相萃取柱凈化、W aters ACQU ITYTMU PLC BEH-C18色譜柱(100mm×2.1mm,1.7μm)分離,在多反應監測模式下進行U PLC-MS/MS分析。7種性激素的方法檢出限(S/N=3)為0.08～0.17μg/kg,定量限(S/N=10)為0.24～0.58μg/kg。考察了內標法和基質匹配外標法對7種性激素進行定量的回收率與精密度:添加水平為1,4μg/kg時,以內標法定量,7種性激素的平均回收率為76%～118%,相對標準偏差(RSD)為5.0%～11.3%;以基質匹配外標法定量,7種性激素的平均回收率為66%～94%,RSD為4.5%～10.7%。該結果表明兩種方法均能夠滿足魚制品中7種性激素的多殘留檢測要求。應用建立的方法對市售脫脂大黃魚及烤魚片進行檢測,未發現7種目標違禁性激素。

超高效液相色譜-串聯質譜法;性激素;多殘留;魚制品;ZnCl2去脂

Abstract:A rap id,specific and highly sensitive method for the determination of seven sexhormones(norgestrel,methyltestosterone,testosterone propionate,medroxy progesterone acetate,megestrol acetate,chlormadinone acetate,and nandrolone)residues in fish p roducts was developed using ultra performance liquid Chromatography-tandem mass spectrometry(U PLCMS/MS)with electrospray ionization(ESI)in positive m ode.The target com pounds were extracted with m ethanol after the enzyme hydrolysis of the fish products.ZnCl2was added to the extract solution to remove lip ids.Then target com pounds were purified by an LC-C18and an LCNH2solid phase extraction cartridges.The target com pounds were separated on a Waters ACQU ITYTMU PLC BEH-C18column(100mm×2.1mm,1.7μm)and detected qualitatively and quantitatively inmulti reaction monitoring(MRM)m ode.For the seven sexhormones,the limits of detection(LOD)of the method were from0.08to0.17μg/kg and the limits of quantification(LOQ)were in the range of0.24-0.58μg/kg.A t the spiked levels of1and4μg/kg,the average recoveries ranged from76%to118%with the relative standard deviations between 5.0%and11.3%for the seven sexhormones using internal standard method;and the average recoveries ranged from66%to94%with the relative standard deviations between4.5%and 10.7%using matrix m atched external standard method.The results show ed that both methods are able to meet the m ulti-residue detection of the seven sex hormone residues in fish products.The degreased large yellow croaker and roast fish fillet real samples from a local market were detected by the developed method,and the seven targets were not found.

Key words:ultra perform ance liquid Chromatography-electrosp ray tandem mass spectrometry(U PLC-MS/MS);sexhormones;multi-residues;fish p roducts;ZnCl2delipidation

食用動物飼養中促生長性激素的使用已經成為全球性的食品安全問題。研究表明,兒童性早熟、婦女乳腺癌和子宮癌發病率上升與動物性食品中性激素殘留有關[1,2],因而歐盟在1996年[3]、我國在2002年[4]相繼頒布法令,禁止合成性激素及具有促生長效用的物質作為促生劑在食用動物飼養中使用?！?br>