HPLC-FLD方法检测早孕期大鼠胚胎的苯并芘水平  

Level of Benzo(a)pyrene Detected by High Performance Liquid Chromatography in Embryonic Tissues of Rat Early Pregnancy

作　　者：梁婧[1] 侯海燕 王梦 孙旸 陈亚琼 LIANG Jing;HOU Hai-yan;WANG Meng;SUN Yang;CHEN Ya-qiong(Guang′anmen South Area Hospital,China Academy of Chinese Medical Sciences,Beijing 102618,China;Department of Obstetrics and Gynecology,Characteristic Medical Center of Chinese People's Armed Police Force,Tianjin 300162,China;Tianjin Key Laboratory for Prevention and Control of Occupational and Environmental Hazards,Tianjin 300162,China)

机构地区：[1]中国中医科学院广安门医院南区,北京102618 [2]中国人民武装警察部队特色医学中心妇产科 [3]天津市职业与环境危害防制重点实验室

出　　处：《国际生殖健康／计划生育杂志》2019年第6期454-458,共5页Journal of International Reproductive Health/Family Planning

基　　金：国家自然科学基金面上项目（81273977）;天津市自然科学基金一般项目（12JCYBJC16200）

摘　　要：目的:建立一种高效液相色谱-荧光检测法(HPLC-FLD),检测给药大鼠早孕胚胎组织中苯并芘(BaP)水平,为其临床应用提供实验依据。方法:采用日本岛津高效液相色谱仪和岛津RF-535荧光检测器,检测给药大鼠早孕胚胎组织中BaP的浓度。样品用乙腈提取处理,无需净化。使用Kromasil C18色谱柱(250mm×4.6 mm,5μL),流动相为甲醇/水(95∶5,V/V);流速为1.0 mL/min;荧光检测器的参数设定:激发波长(Ex)为364 nm,发射波长(Em)为427 nm;柱温为室温;进样量为20μL。结果:BaP在0.002~0.5μg/mL范围内呈良好的线性范围,提取标准曲线的回归方程Y=426 174x-316.95,相关系数0.999 9,本方法的样品提取回收率大于95%,日内差实验的精密度相对标准偏差(RSD)为3.38%,日间差实验的精密度RSD为2.80%,稳定性(n=6)RSD为2.58%,实验检测出样品中BaP的浓度范围在0~0.583 7μg/g。结论:本方法操作简便、实验成本低,准确度、敏感度符合要求,样品前处理过程简单、用时少,可用于给药大鼠早孕胚胎组织中BaP的检测和研究。Objective:To establish a high performance liquid chromatography with fluorescence detection(HPLC-FLD) for the detection of Benzo(a)pyrene in the embryonic tissues of rat early pregnancy, so as to provide experimental data for future clinical application. Methods:The level of Benzo(a)pyrene in the embryonic tissues of rats was determined by Shimadzu HPLC with Shimadzu RF-535 fluorescence detector. The samples were extracted by acetonitrile, without cleaned-up. The chromatographic separation was performed using a column of Kromasil C18(250 mm×4.6 mm, 5 μL) with a mobile phase consisting of methanol and water(95∶5, V/V) at the flow rate of 1.0 mL/min. Excitation wavelength(Ex) of fluorescence detection was 364 nm, the emission wavelength(Em)427 nm, and the injection volume 20 μL. Results:The level of Benzo(a)pyrene in the tissues of rats was linear in the range of 0.002 μg/mL to 0.5 μg/mL. The regression equation of standard curve of extraction was Y=426 174 x-316.95 with a correlation coefficient of 0.999 9. The average recovery for Benzo(a)pyrene standard spiked was more than 95% while the inter-day precision RSD was 3.38%, the intra-day precision RSD was 2.80%, and the stability(n=6) was 2.58%. The benzo(a)pyrene concentration of samples was in the range of 0-0.5837 μg/g. Conclusions:The HPLC-FLD method has the advantages of simple operation, low experiment cost, and coincident veracityand sensitivity. The process of sample extraction is simple and time-saving. We conclude that the HPLC-FLD may be a suitable way for the detection of benzo(a)pyrene in samples such as embryonic tissues in the clinical practice and research projects.

关 键 词：谱法 高压液相 荧光计 苯并芘 胚胎 哺乳动物 妊娠初期 

分 类 号：R28[医药卫生—中药学]