分析的代谢组学变化引起睾酮酯猪血浆和尿液

激素和生长促进剂的使用牲畜养肥的目的已被禁止在欧盟理事会指令93/22 / EC的自1988年以来。然而,这种被禁止的物质仍报道欧洲残监测计划的框架内。为了提高检测方法的可靠性,代谢组学方法一道筛查检测使用自然类固醇激素合成实践或新的合成生长促进剂设计最近[1]。在这项研究中,代谢指纹辨别猪17β-testosteron酯处理和控制动物已被调查。十二个90天猪随机分为测试动物(8)和控制动物(4)组。测试组的动物对待贝聿铭管理激素制剂(丙酸睾酮、苯丙酸睾酮、睾酮isocapronate,睾酮癸酸盐;250年Sustanon喷嘴速度推理法、CZ Reg.56/357/91-C)。收集尿液样本在两组治疗后从14到90天。血浆样本收集从第一天到90天。猪都是每周称重实验。 Samples were filtered on centrifugal devices to remove proteins and obtained filtrates were mixed with internal standard (testosterone-D3 in methanol, Sigma-Aldrich). Each sample was injected into chromatographic system using an Accera 1200 Series on a Hypersil Gold C18 column for separation. LC-HRMS metabolomic fingerprints were acquired on Q-Exactive mass spectrometer (Thermo Fisher Scientific) in ESI+ mode. Full scan mass spectra were acquired from 80 to 800 m/z using a mass resolution of 70.000 FWHM in centroid mode. Raw data were processed by SIEVE and XCMS software. PCA and OPLS-DA were carried out using SIEVE, XCMS and SIMCA-P+ statistic software. The chromatographic analysis of the fingerprints generated 968 ions and 692 ions for plasma and urine, respectively. Statistical evaluation selected only significant peaks (p-value), 374 and 264 peaks were extracted for plasma and urine samples, respectively. Multivariate statistical analysis showed significant metabolic differences between test and control groups on day 28 after aplication of the testosterone hormonal preparation. However, no significant differences between treated and control group were found after day 42. Moreover, the urine samples were examined by conventional target LC-MS/MS. In this case, no testosterone residues were detected after day 14. The study showed also higher growth performance (p<0.05) in treated pigs in comparison to control animals, demonstrating anabolic effect of testosterone esters. The work is going on, additional experimental animals will be used to confirm the fingerprinting data to the control of banned testosterone preparation in pigs.