Stable Isotope Labeling-Based Nontargeted Strategy for Characterization of the In Vitro Metabolic Profile of a Novel Doping BPC-157 in Doping Control by UHPLC-HRMS.
Tian TianJing JingYuanyuan LiYang WangXiaojun DengYuanhong ShanPublished in: Molecules (Basel, Switzerland) (2023)
Traditional strategies for the metabolic profiling of doping are limited by the unpredictable metabolic pathways and the numerous proportions of background and chemical noise that lead to inadequate metabolism knowledge, thereby affecting the selection of optimal detection targets. Thus, a stable isotope labeling-based nontargeted strategy combined with ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) was first proposed for the effective and rapid metabolism analysis of small-molecule doping agents and demonstrated via its application to a novel doping BPC-157. Using 13 C/ 15 N-labeled BPC-157, a complete workflow including automatic 13 C 0 , 15 N 0 - 13 C 6 , 15 N 2 m / z pair picking based on the characteristic behaviors of isotope pairs was constructed, and one metabolite produced by a novel metabolic pathway plus eight metabolites produced by the conventional amide-bond breaking metabolic pathway were successfully discovered from two incubation models. Furthermore, a specific method for the detection of BPC-157 and the five main metabolites in human urine was developed and validated with satisfactory detection limits (0.01~0.11 ng/mL) and excellent quantitative ability (linearity: 0.02~50 ng/mL with R 2 > 0.999; relative error (RE)% < 10% and relative standard deviation (RSD)% < 5%; recovery > 90%). The novel metabolic pathway and the in vitro metabolic profile could provide new insights into the biotransformation of BPC-157 and improved targets for doping control.
Keyphrases
- high resolution mass spectrometry
- ultra high performance liquid chromatography
- liquid chromatography
- ms ms
- gas chromatography
- transition metal
- mass spectrometry
- healthcare
- simultaneous determination
- high resolution
- air pollution
- computed tomography
- label free
- deep learning
- wastewater treatment
- pet imaging
- positron emission tomography
- stress induced