Trichosanthis Radix (TR) is one of the most severely sulfur-fumigated herbs in the market, whose transformation mechanism of chemical compositions and sulfur-fumigation markers of TR have not been clarified. To excavate characteristic sulfur-fumigation markers of TR samples, this study brings up a practical protocol using both ultra-performance liquid chromatography/quadrupole time-of-flight-mass spectrum (UPLC-ESI-QTOF-MS/MS)-based non-targeted metabolomics and ultra-performance liquid chromatography/electrospray ionization/quadrupole multiple-stage linear ion-trap mass spectrum (UPLC-ESI-QTRAP-MS/MS)-based widely targeted metabolomics. The results of study demonstrated that five characteristic markers are sulfur-containing components, which were identified as p-Hydroxybenzyl hydrogen sulfite, cucurbitacin D sulfite I, cucurbitacin D sulfite II, cucurbitacin B sulfite I, and cucurbitacin B sulfite II, respectively. Additionally, cucurbitacin B and D were also filtered and identified as the characteristic sulfur-fumigation markers. Meanwhile, the different sulfur-fumigation extent of TR samples was tested by chemical transformations analysis and sulfur dioxide residues test. Further, 58.16% (139 of 239) of the differential metabolites content significantly reduced in sulfur-fumigated TR samples. Besides, 20 kinds of non-sulfur marker metabolites were tested to evaluate the quality of TR samples before and after sulfur fumigation, predominantly including phenolic acids, amino acids, lipids and nucleotides. Taking TR as an example, this work provides a comprehensive practical protocol for the quality supervision of sulfur-fumigation herbs.

cho虫（Trichosanthis Radix，TR）是市场上最严重的硫熏熏草药之一，其化学成分和硫熏蒸标记物的转化机理尚不清楚。为了挖掘TR样品的特征性硫熏蒸标记，本研究提出了一种基于超高效液相色谱/四极杆飞行时间质谱（UPLC-ESI-QTOF-MS / MS）的非常规方法靶向代谢组学和基于高效液相色谱/电喷雾电离/四极杆多级线性离子阱质谱（UPLC-ESI-QTRAP-MS / MS）的广泛靶向代谢组学。研究结果表明，五个特征性标志物是含硫成分，被确定为p-羟基苄基亚硫酸氢盐，葫芦素D亚硫酸盐I，葫芦素D亚硫酸盐II，葫芦素B亚硫酸盐I和葫芦素B亚硫酸盐II。另外，葫芦素B和D也被过滤并鉴定为特征性硫熏蒸标记物。同时，通过化学转化分析和二氧化硫残留量测试对TR样品的不同硫熏蒸程度进行了测试。此外，硫熏蒸的TR样品中的差异代谢物含量显着降低了58.16％（239个中的139个）。此外，还测试了20种非硫标记代谢物，以评估硫熏蒸前后TR样品的质量，主要包括酚酸，氨基酸，脂质和核苷酸。以TR为例