Evaluation of cellular antioxidant components of honeys using UPLC-MS/MS and HPLC-FLD based on the quantitative composition-activity relationship
Introduction
Honey is a natural substance well known for its various biological activities, e.g. antioxidant (Pichichero et al., 2009, Di Marco et al., 2018, Zhou et al., 2012), antimicrobial (Leyva-Jimenez et al., 2019, Fyfe et al., 2017, Pasias et al., 2018), and anti-inflammatory activities (Kassim et al., 2010, Abdelrahman et al., 2018, Zaidi et al., 2019). Carbohydrates comprise 95% of honey dry weight, especially mono- and di-saccharides such as glucose and fructose in larger quantities. There are also various proteins, minerals, amino acids, phenolic compounds and other minor components in honey, which are closely related to their botanical origin, processing and environmental conditions (Gašić et al., 2014, Chen et al., 2017).
Oxidative stress occurs when the level of oxidation exceeds the antioxidant defenses, which produces oxidative damage that exerts an influence on various physiological functions. Oxidation is caused by reactive oxygen species and free radicals in cellular systems, which are involved in senescence and incidence of cardiovascular disease, diabetes mellitus, and cancer (Rahal et al., 2014). There are various antioxidant molecules, containing phenolic acids, flavonoids and amino acids (Pérez, Iglesias, Pueyo, González, & de Lorenzo, 2007). In most cases, chemical-based methods are applied to evaluate the antioxidant activity of honey, e.g. Ferric reducing antioxidant power (FRAP) assay, 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assay and Oxygen radical absorbance capacity (ORAC) method (Bueno-Costa et al., 2016, Can et al., 2015, Gašić et al., 2014). However, the cellular antioxidant activity assay (CAA) not only quantifies bioactivities of antioxidant compounds in cultured cells, but also comprehensively considers the adsorption, distribution, metabolism, excretion and bioavailability of the antioxidant compounds. So, CAA method is more biologically representative than the chemical-based methods. Although CAA was reported for the antioxidant assessment of various fruits (Wolfe et al., 2008, Wolfe and Liu, 2007), it has not still been applied to the systematic research of honey except only one paper reported CAA for the phenolic extracts of buckwheat and manuka honey (Deng et al., 2018). Actually, more kinds of honey should be investigated for the systematic and comprehensive evaluation of the antioxidant activity of honey. Meanwhile, current available methods for antioxidant study of honey just simply determine the activity or analyze the chemical components. However, considering the antioxidant activity of honey is closely related with the interactions of multiple components, there are still challenges that need to holistically evaluate the cellular antioxidant activity of honeys with different floral and geographic origins, and comprehensively disclose the specific and effective antioxidant components in honey. Therefore, it is necessary to establish a quantitative composition-activity relationship (QCAR) to construct the integrative evaluation, and further discover contributing components in the antioxidant activity of honey (Zhang et al., 2018, Jiang et al., 2012).
In the present research, 39 honey samples from different floral origins (acacia, jujube, vitex, linden, fennel, buckwheat and manuka) were collected to analyze the bioactive compounds using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) with pre-column derivatization. The antioxidant components and activity of fennel honey were firstly investigated. For the first time, CAA assay and QCAR were combined to systematically and comprehensively evaluate the antioxidant markers in honeys, which may facilitate the promotion of the nutritional values and the exploration and utilization of various honeys with different floral and geographic origins.
Section snippets
Honey samples
Acacia, jujube, vitex, linden, fennel, buckwheat, manuka honey samples were acquired from local beekeepers or purchased from manufacturers. All samples were stored at 4 °C. The collection sites were presented in Table 1.
Chemical reagents
Analytical standards 3,4-dihydroxy benzoic acid (98.2%), 4-hydroxy benzoic acid (99.7%), chlorogenic acid (96.0%), caffeic acid (99.2%), syringic acid (96.4%), p-coumaric acid (99.6%), sinapic acid (98%), ferulic acid (99.6%), quercetin (96.0%), quercetrin (98.3%), fisetin
Determination of TPC and TFC
TPC and TFC of 39 honey samples were determined, and the results indicated that TPC ranged from 9.15 (acacia honey) to 294 (buckwheat honey) mg GAE/100 g and TFC ranged from 6.85 (acacia honey) to 64.8 (buckwheat honey) mg QE/100 g, as shown in Table 1. Compared to the highest phenolic content (89.9 mg GAE/100 g) reported in manuka honeys among four floral honeys examined (Alzahrani et al., 2012), buckwheat honeys had the highest phenolic content (61.4–294 mg GAE/100 g), followed by fennel
Conclusion
In the present research, more antioxidant components of phenolic acids, flavonoids and free amino acids in honeys from different floral origins were analyzed using UPLC-MS/MS, HPLC-FLD, and isosakuranetin, cryptochlorogenic acid and methionine were firstly found in honeys. Phenolic acids, flavonoids and free amino acids in honeys varied, relating to the differential antioxidant competences and potential floral markers. CAA assay and QCAR were combined to systematically and comprehensively
Acknowledgements
The research was supported by the Grant from National Natural Science Foundation of China financially (No. 31501477).
Conflict of Interest.
The authors declare no conflict of interest.
References (33)
- et al.
Clove (Syzygium aromaticum) and honey extracts significantly reduce inflammatory cytokines and liver function enzymes in experimental rats fed on carbon tetrachloride (CCl4)
Journal of Radiation Research and Applied Sciences
(2018) - et al.
Pharmacological activities, chemical profile, and physicochemical properties of raw and commercial honey
Biocatalysis and Agricultural Biotechnology
(2019) - et al.
Antibacterial and antioxidant activity of honeys from the state of Rio Grande do Sul, Brazil
LWT - Food Science and Technology
(2016) - et al.
An investigation of Turkish honeys: Their physico-chemical properties, antioxidant capacities and phenolic profiles
Food Chemistry
(2015) - et al.
Biochemical properties, antibacterial and cellular antioxidant activities of buckwheat honey in comparison to manuka honey
Food Chemistry
(2018) - et al.
Compositional analysis of Scottish honeys with antimicrobial activity against antibiotic-resistant bacteria reveals novel antimicrobial components
LWT - Food Science and Technology
(2017) - et al.
Phenolic profile and antioxidant activity of Serbian polyfloral honeys
Food Chemistry
(2014) - et al.
A highly water-soluble disulfonated tetrazolium salt as a chromogenic indicator for nadh as well as cell viability
Talanta
(1997) - et al.
Identification of antitumor constituents in curcuminoids from Curcuma longa L. based on the composition–activity relationship
Journal of Pharmaceutical and Biomedical Analysis
(2012) - et al.
Ellagic acid, phenolic acids, and flavonoids in Malaysian honey extracts demonstrate in vitro anti-inflammatory activity
Nutrition Research
(2010)
Potential antimicrobial activity of honey phenolic compounds against Gram positive and Gram negative bacteria
LWT
Evaluating the antioxidant capacity of natural products: A review on; chemical and cellular-based assays
Analytica Chimica Acta
Antitumour and antioxidant potential of some selected Pakistani honeys
Food Chemistry
Effect of late harvest and floral origin on honey antibacterial properties and quality parameters
Food Chemistry
Antioxidant activity and phenolic composition of herbhoneys
Food Chemistry
Biological properties of phenolic compound extracts in selected Algerian honeys—The inhibition of acetylcholinesterase and α-glucosidase activities
European Journal of Integrative Medicine
Cited by (37)
Sustainable approach for defatted date seed valorization through ultrasonication-based green extraction: A prospective approach for nutraceutical applications
2023, Sustainable Chemistry and PharmacyAn updated review of extraction and liquid chromatography techniques for analysis of phenolic compounds in honey
2022, Journal of Food Composition and AnalysisValidation of the traditional medicinal use of a Mexican endemic orchid (Prosthechea karwinskii) through UPLC-ESI-qTOF-MS/MS characterization of its bioactive compounds
2022, HeliyonCitation Excerpt :In addition, foliar extract reduces insulin resistance, proinflammatory status, and cardiovascular risk in Wistar rats with MS-induced (Barragán-Zárate et al., 2021). Oxidative stress can affect several physiological functions and lead to chronic diseases (Wolfe and Liu, 2007; Shen et al., 2019). ROS are a normal product of metabolism; however, when their concentration exceeds the levels that can be neutralized by endogenous antioxidant systems, oxidative stress occurs.
A comprehensive chemical analysis of New Zealand honeydew honey
2022, Food Research International
- 1
Shi Shen and Jingbo Wang contributed equally to this work.