论文部分内容阅读
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。
Three fertilized eggs of carps (carp, crucian carp and grass carp) were established, and Malachite green (MG) in the ovary and testis and their major metabolite Leucomalachite green (LMG) ) By ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS / MS) detection method. The fertilized egg and ovary were mixed with methanol and extracted with Mcllvaines solution. The MCX solid phase extraction column was used to enrich and purify the testis. After the sperm was extracted by acetonitrile, the n-hexane was degreased. The chromatographic column was equipped with a BEHC18 (1.7μm, 50mm × 2.1mm) column. The mobile phase was acidified with acetonitrile / water and eluted with a gradient of 0.3mL / min. The electrospray ionization-multiple reaction positive ion monitoring mode was used. The results showed that the concentration was between 0.2 ~ 100ng / mL, good linearity, r2≥0.995. The detection limits (LODs) of MG and LMG in fertilized eggs, ovaries and testes were 0.05, 0.05 and 0.10 μg / kg, respectively, and the limits of quantitation (LOQ) were 0.20, 0.20 and 0.30 μg / kg, respectively. The recoveries and relative standard deviations (RSDs) were calculated by the blank matrix addition method. The average recoveries of the testis at the three addition levels of 0.3, 3.0 and 30.0 μg / kg of fertilized eggs and ovaries at 0.5, 5.0 and 50.0 μg / kg were 73.1% ~ 92.5%, the relative standard deviation was 5.3% ~ 12.4%. The method has high sensitivity and good accuracy and is suitable for the determination of MG and LMG residues in the carp zygotes, ovaries and testis.