Department of Nutrition, Exercise and Sports (NEXS), University of Copenhagen, Copenhagen, DENMARK.
Section for Transfusion Medicine, Capital Region Blood Bank, Rigshospitalet, Department of Clinical Medicine, University of Copenhagen, Copenhagen, DENMARK.
Med Sci Sports Exerc. 2021 Jan;53(1):236-243. doi: 10.1249/MSS.0000000000002442.
Autologous blood transfusion is performance enhancing and prohibited in sport but remains difficult to detect. This study explored the hypothesis that an untargeted urine metabolomics analysis can reveal one or more novel metabolites with high sensitivity and specificity for detection of autologous blood transfusion.
In a randomized, double-blinded, placebo-controlled, crossover design, exercise-trained men (n = 12) donated 900 mL blood or were sham phlebotomized. After 4 wk, red blood cells or saline were reinfused. Urine samples were collected before phlebotomy and 2 h and 1, 2, 3, 5, and 10 d after reinfusion and analyzed by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry. Models of unique metabolites reflecting autologous blood transfusion were attained by partial least-squares discriminant analysis.
The strongest model was obtained 2 h after reinfusion with a misclassification error of 6.3% and 98.8% specificity. However, combining only a few of the strongest metabolites selected by this model provided a sensitivity of 100% at days 1 and 2 and 66% at day 3 with 100% specificity. Metabolite identification revealed the presence of secondary di-2-ethylhexyl phtalate metabolites and putatively identified the presence of (iso)caproic acid glucuronide as the strongest candidate biomarker.
Untargeted urine metabolomics revealed several plasticizers as the strongest metabolic pattern for detection of autologous blood transfusion for up to 3 d. Importantly, no other metabolites in urine seem of value for antidoping purposes.
自体输血具有增强运动表现的作用，在运动中被禁止，但仍然难以检测。本研究旨在探索一个假设，即非靶向尿液代谢组学分析是否可以揭示一种或多种具有高灵敏度和特异性的新型代谢物，用于检测自体输血。
采用随机、双盲、安慰剂对照、交叉设计，对运动训练有素的男性（n=12）进行 900ml 采血或模拟采血。4 周后，回输红细胞或生理盐水。采血前、回输后 2h 及 1、2、3、5 和 10d 收集尿液样本，并通过超高效液相色谱-四极杆飞行时间质谱进行分析。采用偏最小二乘判别分析获得反映自体输血的独特代谢物模型。
在回输后 2h 获得的模型最强，错误分类率为 6.3%，特异性为 98.8%。然而，仅结合该模型选择的几个最强代谢物，在第 1、2 和 3 天可达到 100%的灵敏度，特异性为 100%。代谢物鉴定显示存在二-（2-乙基己基）邻苯二甲酸的次级代谢物，推测（异）己酸葡萄糖醛酸苷的存在为最强候选生物标志物。
非靶向尿液代谢组学揭示了几种增塑剂作为检测自体输血的最强代谢模式，最长可持续 3d。重要的是，尿液中没有其他代谢物似乎对反兴奋剂目的有价值。
