【摘要】 建立了3种鲤科鱼(鲤、鲫和草鱼)的受精卵、卵巢及精巢中孔雀石绿(Malachite green,MG)及其在生物体内的主要代谢物隐色孔雀石绿(Leucomalachite green,LMG)的超高效液相色谱串联质谱(UPLC-MS/MS)检测方法。受精卵和卵巢用甲醇混合的Mcllvaines溶液提取,MCX固相萃取柱富集净化;精巢采用乙腈超声提取后正己烷除脂净化。色谱分离柱选用BEHC18(1.7μm,50mm×2.1mm)色谱柱,流动相选用酸化乙腈/水溶液,0.3mL/min梯度洗脱,电喷雾-多反应正离子监测模式。结果表明,浓度在0.2～100ng/mL之间,线性良好,r2≥0.995。MG和LMG在受精卵、卵巢及精巢中的最低检测限(LOD)分别为0.05、0.05和0.10μg/kg,定量限(LOQ)分别为0.20、0.20和0.30μg/kg。以空白基质加入法计算回收率和相对标准偏差,受精卵、卵巢在0.3、3.0和30.0μg/kg3个添加水平下,精巢在0.5、5.0和50.0μg/kg3个添加水平下,平均回收率是73.1%～92.5%,相对标准偏差为5.3%～12.4%。该方法灵敏度高,准确性好,适用于鲤鱼受精卵、卵巢及精巢中MG和LMG残留的测定。更多还原

【Abstract】 A quantitative and qualitative method of ultra performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry(UPLCTM-MS/MS) has been developed for the rapid and sensitive analysis of the malachite green(MG) and its main metabolite leucomalachite green(LMG) in fertilized eggs,ovary and testis in cyprinid fish,such as common carp,crucian and grass carp.Fertilized eggs and ovary samples were extracted by mixture of mcllvaines and methanol and cleaned up using mixed-mode cation exchange solid-phase extraction cartridges(MCX).Then testis sample was extracted with acetonitrile and cleaned up by hexane.Gradient HPLC separation was performed using C18 column(BEH C18 1.7 μm,2.1 mm × 50 mm) with formic acid solution and acetonitrile as the mobile phase.Mass spectrometric determination was carried out with an electrospray ionization interface(ESI) in positive ion mode with multiple reactions monitoring mode(MRM).The calibration curve ranging from 0.2 to 100 ng/mL in fluent phase solvent was used to establish instrument response,with relative standard coefficient beyond 0.995.The method was validated by analysis of eight replicate fertilized eggs and ovary samples fortified with three kinds of spiking concentration levels of 0.3 μg/kg,3.0 μg/kg and 30.0μg/kg MG and LMG,and in testis of 0.50 μg/kg,5.0 μg/kg and 50.0 μg/kg MG and LMG,respectively.Depending on the matrix effect and analytes,relative standard deviations ranged from 5.3% to 12.4%,with average recovery rates around 73.1% to 92.5% in spiked samples.The limit of detection(LOD) and quantification(LOQ) for the analytes in fertilized eggs and ovary were 0.05 μg/kg and 0.10 μg/kg,respectively.And in testis the LOD and LOQ were 0.10 μg/kg and 0.30 μg/kg,respectively.In addition,the method was successfully used to detect the MG and LMG residues in practically aquatic environmental fertilized eggs,ovary and testis in common carp and got a satisfactory result.更多还原