*3.2. Phenolic Composition*

The phenolic composition of haskap berries is presented in Table 3, being identified as constituting one phenolic acid and six anthocyanins. 5-*O*-Caffeoylquinic acid was identified by comparison with a commercial standard and was present in a concentration of 0.589 ± 0.002 mg/g of extract. In other studies involving haskap fruits, other phenolic compounds (non-anthocyanin) were detected, such as protocatechuic, gentisic, ellagic, ferulic, caffeic, and coumaric acids [19]. On the other hand, Oszmiánski et al. [23] found eight phenolic acids, five flavan-3-ols, twelve flavonols, and five flavones. These differences could possibly be explained by the different processing conditions to which the samples were subjected, such as defrosting processes or the maturation stage, climate, harvest, or even storage conditions [2,5,6]. Indeed, several examples can be found for the effect on polyphenolic composition of the different treatments to which the fruits were subjected, for instance, Ochmian et al. [1] packed the berries in polyethylene bags and stored them at −32 ◦C for 6 months, while Khattab et al. [25] stored them for 6 months at −18 ◦C; comparing the results obtained, the samples stored at higher temperatures revealed lower concentrations of phenolic compounds. But besides storage temperature, other factors can affect the phenolic composition; in fact, steam bleaching of the fruits prior to freezing clearly increased the retention of these compounds, whereas storage at −32 ◦C did not reveal such an influence [25]. There are other reports on the pre-treatment of these berries that also report the contribution of the process to the variation of the phenolic composition; for example, in a study performed by Wojdyło et al. [21], the fruits were cut directly into liquid nitrogen and freeze-dried before grounding and storing at −70 ◦C, which also led to the preservation of such compounds [21]. Besides, the influence of the maturation stage of these fruits also revealed a clear influence on the polyphenolic composition, with late harvest haskap presenting significantly higher concentrations than early-harvested fruits [1]; more specifically, for *Wojtek* cultivar, the early berries presented lower contents (174.95 mg/100 g) than the late-harvest berries, in which an increase of 27% was verified. For the *Brazowa* cultivar, similar observations were made, with an enhancement of 65% of the initial concentration (144.84 mg/100 g) [26,27].

Regarding anthocyanins (Table 3), these were identified according to the peak characteristics, such as retention time, wavelength of maximum absorption and mass spectral data. Three cyanidin derivatives (cyanidin-*O*-hexoside-*O*-hexoside, cyanidin-3-*O*-glucoside, and cyanidin-*O*-rhamnoside- *O*-hexoside), two peonidin derivatives (peonidin-3-*O*-glucoside and peonidin-*O*-rhamnoside-*O*-hexoside), and a pelargonidin derivative (pelargonidin-3-*O*-glucoside) were identified, as shown in Figure 1. The compounds represented by peaks two, four, and five were positively identified by comparison with commercial standard chromatographic characteristics. The compound corresponding to peak 1 ([M]<sup>+</sup> at *<sup>m</sup>*/*<sup>z</sup>* 611) presented two MS<sup>2</sup> fragments, presenting two losses of hexosyl units (*m*/*<sup>z</sup>* at 287; <sup>−</sup>162–162 u), being identified as cyanidin-*O*-hexoside-*O*-hexoside. Also, the compounds represented by peaks three ([M]<sup>+</sup> at *m*/*z* 595) and six ([M]<sup>+</sup> at *m*/*z* 609) were identified as cyanidin-*O*-rhamnoside-*O*-hexoside and peonidin-*O*-rhamnoside-*O*-hexoside, respectively, revealing a loss of one hexosyl (−162u) and rhamnosyl (−146u). The most abundant anthocyanin was cyanidin-3-*O*-glucoside, representing 63% of the total anthocyanin concentration (61.7 ± 0.1 of 97.9 ± 0.2 mg/g of extract). The results obtained are in accordance with those reported by Oszmia ´nski et al [23]., but in this study, cyanidin-3-*O*-glucoside represented 88% of the total content, and another seven anthocyanins were found. Ochmian et al. [1] and Wojdyło et al. [21] also reported this anthocyanin as the one present in the highest concentration (83–90% and 71–89% of the total concentration).



0.9986; LOD: 9.94; LOQ: 30.13); 3

LOD: 12.90; LOQ: 39.09).

Pelargonidin-3-*O*-glucoside

 (*<sup>y</sup>* = 61493*x* − 628875; *R*2: 0.9957; LOD: 24.94; LOQ: 75.580.); and 4

Peonidin-3-*O*-glucoside

 (*<sup>y</sup>* = 151438*x* − 3E+06; *R*2: 0.9977;

**Figure 1.** Chromatographic profile of the phenolic compounds found in the haskap fruit hydroethanolic extract, recorded at 520 nm (1: cyanidin-*O*-hexoside-*O*-hexoside; 2: cyanidin-3-*O*-glucoside; 3: cyanidin-*O*-rhamnoside-*O*-hexoside; 4: pelargonidin-3-*O*-glucoside; 5: peonidin-3-*O*-glucoside; and 6: peonidin-*O*-rhamnoside-*O*-hexoside).
