Phytochemical analysis and safety evaluation of the extracts from Tetrastigma hemsleyanum Diels et Gilg leaves

As the market of Tetrastigma hemsleyanum Diels et Gilg (TH) products rapidly expands and consumer demand increases, identifying the chemical components and evaluating its safety can help ensure human health during their usage. In this study, GC-MS and LC-MS were used to investigate the phytochemical characteristics of the TH leaf extracts, and the safety of TH extracts was evaluated using zebrafish embryos and larvae, with specific endpoints including the hatching rate, mortality rate, and the overall morphology. It revealed that TH leaves were rich in vitamins, flavonoids, terpenoids, etc. A total of 31 and 42 metabolites was identified in the n-hexane extract and the 60 % methanol aqueous extract of TH leaves, respectively, using GC-MS and LC/Q-TOF-MS techniques. DL-alpha-tocopherol was discovered for the first time in TH leaves. Seven exposure levels (0.02–6.4 mg/mL) of the aqueous extracts were employed in safety evaluation using zebrafish embryos. Our results indicated that the highest concentration (6.4 mg/mL) led to the lowest hatching rate (1.3 ± 1.8 %, P < 0.001) and the highest cumulative mortality rate (22.5 ± 9.4 %, P < 0.001). Subsequently, five exposure levels (0.02–1.6 mg/mL) were employed to evaluate the safety using zebrafish larvae. The results indicated that the treatment had no significant impact on the mortality rate, overall morphology, or specific organs (heart, liver, kidneys) of zebrafish larvae, nor did it induce morphological abnormalities. However, a concentration-dependent increase in heart rate was observed. This study provides a reference for consumers to use TH leaf safely and rationally.