Determination and confirmation of selective estrogen receptor modulators (SERMs), anti-estrogens and aromatase inhibitors in bovine and porcine urine using UHPLC-MS/MS

Selective estrogen receptor modulators (SERMs), anti-estrogens and aromatase inhibitors are prohibited in human sports doping. However, they also present a risk of being used illegally in animal husbandry for fattening purposes. A method was developed and validated using UHPLC-MS/MS for the determination and confirmation of SERMs, anti-estrogens and aromatase inhibiters in bovine and porcine urine. This method was used in a survey of more than 200 bovine and porcine urine samples from Dutch farms. In 18 out of 103 porcine urine samples (17%) and two out of 114 bovine samples (2%) formestane, an aromatase inhibitor, was detected. None of the other compounds was detected. From human doping control it is known that formestane can, in some cases, be of natural origin. Analyses of reference samples from untreated bovine and porcine animals demonstrated the presence of formestane in bovine animals, but not yet in porcine animals. Future research will focus on whether the detected formestane in porcine and bovine urine is from endogenous or exogenous origin, using GC-c-IRMS.

选择性雌激素受体调节剂（Selective estrogen receptor modulators, SERMs）、抗雌激素类化合物与芳香化酶抑制剂，均被禁止应用于人类体育兴奋剂领域。然而，这类物质同样存在被非法用于畜牧养殖以实现育肥目的的风险。本研究开发并验证了一种采用超高效液相色谱-串联质谱（UHPLC-MS/MS）的分析方法，可对牛尿与猪尿中的SERMs、抗雌激素类化合物及芳香化酶抑制剂进行测定与确证。研究人员采用该方法对来自荷兰农场的200余份牛尿与猪尿样本开展了调研检测。在103份猪尿样本中有18份（占比17%）、114份牛尿样本中有2份（占比2%）检出了作为芳香化酶抑制剂的福美坦（formestane），其余受试化合物均未检出。根据人类兴奋剂管控领域的已有研究成果，福美坦在部分情况下可来源于生物体内源性合成。对未施加任何处理的牛、猪空白参照样本的分析结果显示，牛体内可检出内源性福美坦，但暂未在猪体内发现该物质。后续研究将采用气相色谱-燃烧-同位素比值质谱（GC-C-IRMS）技术，探究猪尿与牛尿中检出的福美坦究竟来自内源性还是外源性来源。