Separating and identifying multiple structural isomers of 3-hydroxy-3-(3′-hydroxyphenyl)propanoic acid (3,3′-HPHPA)

3-hydroxy-3-(3′-hydroxyphenyl)propanoic acid (CID: 102959) (3,3’-HPHPA) is one of several phenolic molecules that are generated by a process involving gut microbiota and have been implicated in autism and schizophrenia. The structural isomers of 3,3′-HPHPA have been challenging to separate chromatographically. Accordingly, their distribution across brain and other compartments in humans and experimental animals remains poorly characterized. We describe a modified liquid chromatography mass spectroscopy method for the detection and separation of seven commercially available structural isomers of 3,3′-HPHPA. We used the method to screen different compartments in the mouse and demonstrate its utility in identifying multiple structural isomers and conjugates of 3,3′-HPHPA in biological matrices. Characterizing the diversity and complexity of this family of structural isomers will advance studies of their bioactivity and clinical relevance.

3-羟基-3-(3'-羟基苯基)丙酸（CID: 102959）（3,3'-HPHPA）属于一类经肠道菌群代谢产生的酚类分子之一，且已被证实与孤独症（自闭症）及精神分裂症存在关联。3,3'-HPHPA的结构异构体在色谱分离方面始终存在较大挑战，因此目前对于该类异构体在人类及实验动物的大脑与其他生物基质中的分布情况仍研究不足。本研究开发了一种改良型液相色谱-质谱联用法，可实现7种商用3,3'-HPHPA结构异构体的分离与检测。我们利用该方法对小鼠体内的不同生物基质进行了筛查，证实其可有效在生物样本中识别出多种3,3'-HPHPA的结构异构体及其结合物。对该类结构异构体家族的多样性与复杂性进行解析，将有助于推动其生物活性及临床相关性相关研究的进展。