Ziziphi Spinosae Semen (ZSS) is a traditional Chinese medicine used for sedation and hypnosis. Preliminary studies have shown that frying it could increase its sedative and hypnotic effects due to an increase in its chemical contents. However, the correlation between increased ZSS contents and therapeutic effects remains unclear. This study aimed to identify chemical components that change between ZSS and Fried Ziziphi Spinosae Semen (FZSS) and Q-markers related to these changed components’ sedative and hypnotic effects. Differences between ZSS and FZSS were investigated using the UPLC fingerprint analysis. Components significantly different between ZSS and FZSS were screened using the UPLC-Q-TOF-MS analysis combined with a multivariate statistical method. In addition, ZSS and FZSS extracts were treated with diazepam in vitro to observe their differences in saturation competition between ZSS extract and diazepam, before and after processing, and diazepam on the GABA receptor in SD rats’ brain tissue. Then, the chemical components of ZSS and FZSS that competed with diazepam to bind to the GABA receptor were identified by LC-MS/MS analysis. Finally, the binding efficiency of the different medicinal components was assessed using molecular docking technology. The results indicated significant differences in the content of various chemical components between ZSS and FZSS. Among them, the contents of adenosine, spinosin, 6′″-feruloylspinosin, jujuboside A and betulinic acid were found to be significantly increased after frying. LC-MS/MS and molecular docking analysis screened spinosin, 6′″-feruloylspinosin and betulinic acid as Q markers for the sedative and hypnotic effects of ZSS and FZSS. In summary, this study identified the changed sedative-hypnotic chemical components and Q-markers of ZSS before and after frying.

Ziziphi Spinosae Semen（ZSS）是一种用于镇静和催眠的中药。初步研究表明，由于其化学成分的增加，油炸它可以增加其镇静和催眠作用。然而，ZSS含量增加与治疗效果之间的相关性仍不清楚。本研究旨在确定 ZSS 和炒紫荆精 (FZSS) 之间变化的化学成分以及与这些变化成分的镇静和催眠作用相关的 Q 标记。使用 UPLC 指纹分析研究了 ZSS 和 FZSS 之间的差异。使用 UPLC-Q-TOF-MS 分析结合多变量统计方法筛选 ZSS 和 FZSS 之间显着差异的成分。此外，ZSS 和 FZSS 提取物用地西泮处理体外观察ZSS提取物与地西泮、加工前后、地西泮对SD大鼠脑组织GABA受体饱和竞争的差异。然后，通过LC-MS/MS分析鉴定了与地西泮竞争结合GABA受体的ZSS和FZSS的化学成分。最后，使用分子对接技术评估不同药物成分的结合效率。结果表明ZSS和FZSS之间各种化学成分的含量存在显着差异。其中腺苷、棘苷、6''-阿魏酰棘苷、大枣苷A和桦木酸的含量在油炸后显着升高。LC-MS/MS 和分子对接分析筛选出了多杀菌素，6'''-阿魏酰斯皮诺辛和桦木酸作为 ZSS 和 FZSS 镇静催眠作用的 Q 标志物。总之，本研究确定了油炸前后 ZSS 镇静催眠化学成分和 Q 标志物的变化。