Xiaofan Yuan

Chlorogenic acid (CGA) is a natural compound with potent antioxidant and antibacterial properties that can be extracted from Eucommia leaves (EL).

In the present study, we investigated the protection of hemp seed polyphenols on human umbilical vein endothelial cells (HUVEC) from H ₂ O ₂ ‐mediated oxidative stress injury. Fractions with different polarities were obtained by separating the hemp seed extract using HPD300 macroporous resin‐packed column. The fraction, desorbed by 50% ethanol, is rich in polyphenol (789.51 ± 21.92 mg GAE/g) and has the highest antioxidant activity in vitro. HPLC‐QTOF‐MS/MS identified the main polyphenol components in hemp seed shells: 4 hydroxycinnamic acid amides and 15 lignanamides. The protective effects of hemp seed polyphenol against oxidative‐stress injury in HUVEC cells were evaluated by cell viability, intracellular antioxidant parameters, and cell apoptosis assay. After HUVEC cells were precultured with 50 µg/ml hemp seed polyphenols, the cell viability increased significantly from 53.07 ± 2.46% (model group) to 80.65 ± 1.32% ( p < 0.01). In addition, the pretreatment of HUVEC cells with polyphenol could substantially increase their intracellular superoxide dismutase activity and reduce their intracellular reactive oxygen species level, malondialdehyde content, and lactate dehydrogenase leakage index. These findings demonstrate the defensive potential of hemp seed polyphenol in reducing the incidence of cardiovascular disease. Practical Application: Hemp seed shell waste is produced while producing hemp seed kernel and has abundant phenolic compounds. This research showed that hemp seed polyphenol has potent antioxidant activity in vitro and protects HUVEC cells against H ₂ O ₂ ‐induced oxidative stress injury, suggesting that hemp seed polyphenol has the defensive potential to reduce the incidence of cardiovascular disease. These results indicated that polyphenol separated from hemp seed shells is valuable for further research and development, which will improve the utilization rate of hemp seed.

An efficient method for simultaneous detoxification and preparative separation of chlorogenic acid from Eupatorium adenophorum was developed. The entire procedure consisted of two separation steps. The first step was enrichment of chlorogenic acid from the crude extract by NKA-II macroporous resin. The purity of chlorogenic acid increased from 5.3% to 20.9% with a recovery of 88.7%. Subsequently, polyamide resin was used to achieve a higher purity. After the polyamide resin treatment, the main toxin 9-oxo-10,11-dehydroageraphorone was removed and the purity of chlorogenic acid reached 86.5% with a recovery of 84.8%. It provided a new way for the utilization of E. adenophorum.

Eupatorium adenophorum can induce liver toxicity in animals. For the safe utilisation of the weed, the hepatotoxic components need to be discovered. In this study, in vitro hepatotoxicity of different extracts from E. adenophorum were determined on human hepatocyte cell line L02 and hepatocellular carcinoma cell line HepG2. The results showed that water extracts of E. adenophorum exhibited no hepatotoxicity in vitro while high concentrations of the organic solvent extracts had obvious hepatotoxicity. Sesquiterpenes may contribute to the toxicity based on the comparison of composition analysis. Three cadinene sesquiterpenes were purified and identified as 9-oxo-10,11-dehydroageraphorone, 10Hα-9-oxo-ageraphorone and 10Hβ-9-oxo-ageraphorone. In vitro hepatotoxic effects of these components were investigated, the IC₅₀ of the three compounds were 122.53, 87.52, and 108.80 μM in L02 cells and 151.92, 104.48, and 138.08 μM in HepG2 cells by Cell Counting Kit-8 (CCK-8) assay. The three components were confirmed to be, at least partial, hepatotoxic components.

Eupatorium adenophorum is widely distributed throughout the world’s tropical and temperate regions. It has become a harmful weed of crops and natural environments. Its leaves contain bioactive compounds such as chlorogenic acid and may be used as feed additives. In this study, chlorogenic acid was extracted and separated from leaves of E. adenophorum. Three chlorogenic acid products were prepared with different purities of 6.11%, 22.17%, and 96.03%. Phytochemical analysis demonstrated that the main toxins of sesquiterpenes were almost completely removed in sample preparation procedure. The three products were evaluated for safety via in vitro and in vivo toxicological studies. All the products exhibited no cytotoxic effects at a dose of 400 μg/mL in an in vitro cell viability assay. When administered in vivo at a single dose up to 1.5 g/kg bw, all three products caused no signs or symptoms of toxicity in mice. These results encourage further exploration of extracts from E. adenophorum in feed additive application.