UHPLC-DAD/ESI-TOF-MS Phytochemical Characterization and Evaluation of the Impact of Eleutherococcus senticosus Fruit Intractum on Biochemical, Hepatological, and Blood Parameters in Balb/c Mice

Abstract

Eleutherococcus senticosus (Rupr. et Maxim.) Maxim. (ES) has gained popularity for its adaptogenic, immunostimulant, and anti-inflammatory properties. Because of overexploitation of the roots, the species is considered to be endangered and has been put on the Red List in some countries (e.g., the Republic of Korea). Therefore, the fruits of E. senticosus might be explored as a new sustainable source of compounds with adaptogenic activity. This study aimed to assess the chemical composition and the safety profile (hepatotoxicity, blood morphology, biochemical parameters of blood plasma) of E. senticosus fruit intractum in Balb/c mice after oral administration of 750 and 1500 mg/kg b.w. UHPLC analysis coupled with DAD and MS detectors was used to quantify the metabolites. For the first time, oleanolic and ursolic acids were quantified in the intractum (16.01 ± 1.3 and 2.21 ± 0.17 µg/g of oleanolic and ursolic acids, respectively). Regarding polyphenols, chlorogenic acid (0.92 mg/g of dried extract), caffeic acid (0.43 mg/g), dicaffeoylquinic acids (in total: 1.27 mg/g), and an unidentified caffeic acid ester (0.81 mg/g) were identified. The results in Balb/c mice revealed that the intractum does not cause significant variations in red blood cells parameters. In turn, a significant decrease in the total number of leukocytes was observed (5.8 × 10³ µL), with a percentage increase in lymphocytes among the groups (80.2, 81.8, and 82.6). The ability of the intractum to decrease alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels may indicate its anti-inflammatory activity. Our observations justify that the fruits of E. senticosus are safe in the doses used and do not cause significant changes in the activity of the liver enzymes or in blood parameters.

Keywords: Balb/c mice; Eleutherococcus senticosus; hepatotoxicity; intractum; triterpenic acids.

Figure 1

The chemical structures of eleutheroside B (syringin 4-β-D-glucoside), eleutheroside E ((−)-siringaresinol 4,4”-O-β-D-diglucoside), and eleutheroside E1 ((−)-siringaresinol 4-O-β-D-glucoside).

Figure 2

Base peak chromatogram (grey line) and the chromatogram registered at λ = 320 nm (red line) of E. senticosus fruit intractum.

Figure 3

The change in average body weight (g) in groups of mice treated with different doses of intractum (mg/kg b.w.).

Figure 4

Average weight of individual organs collected from groups of mice; results presented as mean ± SD. Different letters (a, b) indicate statistically significant differences, with p < 0.05.

Figure 5

H&E staining. Microphotographs of representative liver sections obtained from control mice (A) and animals treated with 750 (B) or 1500 mg/kg b.w. (C) of the intractum.

Figure 6

Hierarchical cluster analysis of three analyzed groups of mice based on all examined parameters.

Figure 7

Principal component analysis of analyzed groups of mice based on all examined parameters.