Evaluation of Hypolipidemic and Antioxidant Effects in Phenolrich Fraction of Crataegus pinnatifida Fruit in Hyperlipidemia Rats and Identification of Chemical Composition by Ultra-performance Liquid Chromatography Coupled with Quadropole Time-of-flight Mass Spectrometry

Abstract

Background: Hawthorn (Crataegus pinnatifida) fruit has enjoyed a great popularity as a pleasant-tasting food associated with hypolipidemic and antioxidant effects.

Objective: Our aim was to screen the effective fraction of hawthorn fruit in the treatment of hyperlipidemia rats. Materials and Methods: In this study, ethanol extract of hawthorn fruit (Fr.1) and four fractionated extracts (Fr.2, Fr.3, Fr.4, and Fr.5) were compared to total phenol content evaluated using Folin-Ciocalteu method, and hypolipidemic and antioxidant effects were assessed in hyperlipidemic rats.

Results: Total phenol content of Fr.4 was higher than other fractions by at least 2 fold. Furthermore, this fraction possessed the strongest hypolipidemic and antioxidant effects in hyperlipidemic rats. On this basis, 15 phenolic compounds and four organic acids in Fr.4 were positively or tentatively identified using ultra-performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry. In addition, 5-O-caffeoyl quinic acid butyl ester was first reported in hawthorn fruit.

Conclusion: Phenol-rich fraction in hawthorn fruit exhibited satisfactory hypolipidemic and antioxidant effects, and this could be exploited for further promotion of functional foods.

Summary: Phenol-rich fraction in hawthorn fruit possesses most potent hypolipidemic and antioxidant effects in hyperlidemia rats. Abbreviations used: UPLC-Q-TOF-MS/MS: Ultra performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry; TC: Total cholesterol; TG: Triglyceride; LDL-C: Low-density lipoprotein-cholesterol; HDL-C: High-density lipoprotein-cholesterol; GSH-Px: Glutathione peroxidase; SOD: Superoxide dismutase; MDA: Malondialdehyde; CAT: Catalase; NO: Nitric oxide; NOS: Nitric oxide synthase; ROS: Reactive oxygen species; •OOH: Superoxide anions, •OH: Hydroxyl radicals.

Keywords: Antioxidant effect; Crataegus pinnatifida fruit; hypolipidemic effect; phenolic compounds; ultra performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry.

Figure 1

Preparation procedure of the fractions of hawthorn fruit

Figure 2

Effects of Crataegus pinnatifida fruit fractions on total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol in rats fed on normal diet or high-fat emulsion diet. The low-, medium-, and high-doses were equivalent to about 75, 150, and 300 mg/kg body weight, respectively. Values are shown as mean ± standard deviation (n = 8). ^#P < 0.05 and ^##P < 0.01 compared with the control group. *P < 0.05 and **P < 0.01 compared with the model group

Figure 3

Effects of Crataegus pinnatifida fruit on malondialdehyde, nitric oxide content, and the activities of catalase, superoxide dismutase, glutathione peroxidase, and nitric oxide synthase of serum in rats fed on high-fat emulsion or normal diet. The low-, medium-, and high-doses were equivalent to about 75, 150, and 300 mg/kg body weight, respectively. Values are shown as mean ± standard deviation (n = 8). ^#P < 0.05 and ^##P < 0.01 compared with the control group. *P < 0.05 and **P < 0.01 compared with the model group

Figure 4

Effects of Crataegus pinnatifida fruit on malondialdehyde content and the activities of superoxide dismutase, catalase, and glutathione peroxidase of liver in rats fed on normal diet or high-fat emulsion diet. The low-, medium-, and high-doses were equivalent to about 75, 150, and 300 mg/kg body weight, respectively. Values are shown as mean ± standard deviation (n = 8). ^#P < 0.05 and ^##P < 0.01 compared with the control group. *P < 0.05 and **P < 0.01 compared with the model group

Figure 5

Total ion chromatogram of Fr.4 analyzed by ultra-performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry

Figure 6

Detailed fragmentation pathways and characteristic ions of peak 19