Bioavailability of anthocyanins and colonic polyphenol metabolites following consumption of aronia berry extract
Introduction
Aronia berries are a rich source of dietary polyphenols, including anthocyanins, hydroxycinnamic acids and proanthocyanidins (Taheri, Connolly, Brand, & Bolling, 2013). Studies have suggested that aronia berry consumption improves dyslipidemia, inhibits inflammation, and reduces oxidative stress associated with chronic diseases in humans (Broncel et al., 2010, Naruszewicz et al., 2007, Porȩba et al., 2009, Skoczyńska et al., 2007) and animals (Kim et al., 2013, Kujawska et al., 2011, Skoczyńska et al., 2007, Valcheva-Kuzmanova et al., 2007). Polyphenols are extensively metabolized and have apparent low bioavailability (Manach, Scalbert, Morand, Rémésy, & Jiménez, 2004). Therefore, further data are needed to reconcile aronia polyphenol bioavailability with its apparent health effects.
The most abundant aronia polyphenols, i.e., anthocyanins and proanthocyanins, have less than 6% bioavailability of the initial dose ingested (Cao et al., 2001, Miyazawa et al., 1999, Wu et al., 2002). When considering colonic metabolites, the bioavailability of anthocyanin and proanthocyanidins may range from 12% to 18% (Czank et al., 2013, Gonthier et al., 2003). Anthocyanins can be absorbed in the intestine, and further subjected to phase II metabolism in the gut or liver (Kay, 2006, Wu et al., 2002). Also, spontaneous degradation or microbial catabolism of anthocyanins may lead to the formation of phenolic acids, such as protocatechuic acid (Czank et al., 2013). Aronia proanthocyanidins are primarily polymeric, which limits absorption to its microbial catabolites (Déprez et al., 2000, Ou and Gu, 2014). These catabolites include valerolactones, hydroxyphenyl propionic acids, hydroxyphenyl acetic acids, and hydroxybenzoic acids that are absorbed from the colon (Rios et al., 2003, Urpi-Sarda et al., 2009).
The bioavailability and excretion of aronia anthocyanins and its phase II metabolites following juice and extract consumption has been previously described (Kay et al., 2005, Kay et al., 2004, Wiczkowski et al., 2010, Woodward et al., 2009). However, the bioavailability of major colonic catabolites has not been well-defined. The catabolism of aronia polyphenols converges on several key intermediates, including hippuric acid, protocatechuic acid, and phenylproprionic acids (Fig. 1) (El Mohsen et al., 2006, Monagas et al., 2010, Stalmach et al., 2013, Wu et al., 2002, Wu et al., 2004). Therefore, the objectives of this study were to quantitate the bioavailable aronia anthocyanins and colonic polyphenol catabolites in humans following consumption of aronia extracts. Furthermore, we described the validation of analytical methods used to generate these data.
Section snippets
Reagents and materials
Methanol was LC-MS grade from Fischer Scientific (Fair Lawn, NJ, USA). Anthocyanin standards with ⩾98% purity were purchased from Wuxi App Tec Co. (Shanghai, China). All other chemicals and reagents, including LC-MS grade formic acid and trifluoroacetic acid, were acquired from Sigma-Aldrich (St. Louis, MO, USA). Aronia berry extract was purchased from Artemis International (Fort Wayne, IA, USA) and packaged into opaque cellulose capsules containing 250 mg extract ±10% per capsule (Beehive
Polyphenol content of aronia extract supplement
The values are reported in the (Table 2). Anthocyanins and proanthocyanins were the most abundant polyphenol classes in aronia extract, accounting for 34% and 31% of the total phenolic content, respectively. Anthocyanins were primarily cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, cyanidin-3-O-arabinoside, and cyanidn-3-O-xyloside. Proanthocyanidins were 0.2% monomers, 7.1% dimers, 32.2% trimers, and 60.4% degree of polymerization (DP) > 4. In total, a 500 mg dose of aronia berry extract
Discussion
The UHPLC-MS methods developed for polyphenol analysis in plasma and urine had acceptable reproducibility and sensitivity. Routine RSD was ⩽10% without including an internal standard in the method. Including a stable isotope standard would be expected to further improve precision. The recovery rates after SPE of plasma polyphenols were 10–20% higher than previously reported, which was around 70–90% for other anthocyanin-rich foods (Wiczkowski et al., 2010, Zhang and Zuo, 2004). However, the SPE
Conflicts of interest
The authors declare no conflicts of interest.
Acknowledgement
This work was funded by the Connecticut Department of Public Health.
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