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. 2022 Aug 20;11(8):1618.
doi: 10.3390/antiox11081618.

A Comparative Study of Hesperetin, Hesperidin and Hesperidin Glucoside: Antioxidant, Anti-Inflammatory, and Antibacterial Activities In Vitro

Sung-Sook Choi  1   2 , Sun-Hyung Lee  1 , Kyung-Ae Lee  3
Affiliations

A Comparative Study of Hesperetin, Hesperidin and Hesperidin Glucoside: Antioxidant, Anti-Inflammatory, and Antibacterial Activities In Vitro

Sung-Sook Choi et al. Antioxidants (Basel). .

Abstract

The antioxidant, anti-inflammatory and antibacterial activities of hesperetin, hesperidin and hesperidin glucoside with different solubility were compared in vitro. Hesperetin was prepared by enzymatic hydrolysis from hesperidin, and hesperidin glucoside composed of hesperidin mono-glucoside was prepared from hesperidin through enzymatic transglycosylation. Solubility of the compounds was different: the partition coefficient (log P) was 2.85 ± 0.02 for hesperetin, 2.01 ± 0.02 for hesperidin, and -3.04 ± 0.03 for hesperidin glucoside. Hesperetin showed a higher effect than hesperidin and hesperidin glucoside on radical scavenging activity in antioxidant assays, while hesperidin and hesperidin glucoside showed similar activity. Cytotoxicity was low in the order of hesperidin glucoside, hesperidin, and hesperetin in murine macrophage RAW264.7 cells. Treatment of the cells with each compound reduced the levels of inflammatory mediators, nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6). Hesperetin was most effective at relatively low concentrations, however, hesperidin glucoside was also effective at higher concentration. Hesperetin showed higher antibacterial activity than hesperidin in both Gram-positive and -negative bacteria, and hesperidin glucoside showed similarly higher activity with hesperetin depending on the bacterial strain. In conclusion, hesperetin in the form of aglycone showed more potent biological activity than hesperidin and hesperidin glucoside. However, hesperidin glucoside, the highly soluble form, has been shown to increase the activity compared to poorly soluble hesperidin.

Keywords: anti-inflammatory; antibacterial; antioxidant; hesperetin; hesperidin; hesperidin glucoside; in vitro; solubility.

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Conflict of interest statement

The authors have no conflicts of interest to declare.

Figures

Figure 1
Figure 1
Structures of hesperidin and related compounds. (A) hesperetin, (B) hesperidin and (C) hesperidin glucoside (mono-glucoside).
Figure 2
Figure 2
HPLC profiles of hesperidin glucosides synthesized during enzymatic transglycosylation. The x- and y-axis represents the retention time (min) and arbitrary unit (AU), respectively. Abbreviation, HD; hesperidin, HG1; hesperidin mono-glucoside, HG2; hesperidin di-glucoside, HG3; hesperidin tri-glucoside, HG4; hesperidin tetra-glucoside, HG5; hesperidin penta-glucoside.
Figure 3
Figure 3
FT-IR spectra of hesperetin, hesperidin and hesperidin glucosides. (A) Hesperetin, (B) Hesperidin, (C) hesperidin glucoside (mono-glucoside).
Figure 4
Figure 4
Mass spectra and fragmentation of prepared hesperetin, hesperidin and hesperidin glucoside by LC-MS-(MS) analysis. (A) Spectra of hesperetin, (B) spectra of hesperidin, (C) spectra of hesperidin glucoside.
Figure 5
Figure 5
Antioxidant activity of hesperetin, hesperidin and hesperidin glucoside. Results were expressed as a mean ± SD (n = 3). (A) 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, (B) 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging activity. CTR+; positive control (ascorbic acid), HT; hesperetin, HD; hesperidin, HG; hesperidin glucoside.
Figure 6
Figure 6
Effect of hesperetin, hesperidin and hesperidin glucoside on RAW 264.7 cell viability. Cells were treated with hesperetin, hesperidin and hesperidin glucoside, respectively, for 1 h and incubated for 24 h with LPS (final concentration of 1 μg/mL. Results were represented as mean ± SD (n = 3). Significant differences from the control (** p < 0.01). CTR; Control (with LPS), HT; hesperetin, HD; hesperidin, HG; hesperidin glucoside.
Figure 7
Figure 7
Effect of hesperetin, hesperidin and hesperidin glucoside on the production of nitric oxide (NO), Prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6). RAW 264.7 cells were cultured with different concentrations of hesperetin, hesperidin and hesperidin glucoside for 24 h with LPS (1 μg/mL). Results were represented as mean ± SD (n = 3). (A) Effect on NO levels; (B) Effect on PGE2 levels; (C) Effect on TNF-α levels; (D) Effect on IL-6 levels. Different letters (a, b, c, d) above the bars indicate significant differences (p < 0.05), where letters ab or bc indicate the intermediate significance between a and b, or b and c, respectively. CTR−; Negative control, CTR+; Positive control (with LPS), HT; hesperetin, HD; hesperidin, HG; hesperidin glucoside.
Figure 8
Figure 8
An example of precipitates containing microbial cells and insolubilized compounds in culture broth.
See this image and copyright information in PMC

References

    1. Németh K., Plumb G.W., Berrin J.-G., Juge N., Jacob R., Naim H.Y., Williamson G., Swallow D.M., Kroon P.A. Deglycosylation by small intestinal epithelial cell β-glycosidases is a critical step in the absorption and metabolism of dietary flavonoid glycosides in humans. Eur. J. Nutr. 2003;42:29–42. - PubMed
    1. Matsumoto H., Ikoma Y., Sugiura M., Yano M., Hasegawa Y. Identification and quantification of the conjugated metabolites derived from orally administered hesperidin in rat plasma. J. Agric. Food Chem. 2004;52:6653–6659. - PubMed
    1. Pereira-Caro G., Polyviou T., Ludwig I.A., Nastase A.M., Moreno-Rojas J.M., Garcia A.L., Malkova D., Crozier A. Bioavailability of orange juice (poly)phenols: The impact of short-term cessation of training by male endurance athletes. Am. J. Clin. Nutr. 2017;106:791–800. - PubMed
    1. Ávila-Gálvez M.Á., Giménez-Bastida J.A., González-Sarrías A., Espín J.C. New Insights into the Metabolism of the Flavanones Eriocitrin and Hesperidin: A Comparative Human Pharmacokinetic Study. Antioxidants. 2021;10:435. - PMC - PubMed
    1. Cho J. Antioxidant and neuroprotective effects of hesperidin and its aglycone hesperetin. Arch. Pharmacal. Res. 2006;29:699–706. - PubMed

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