Distribution of phytochemicals and antioxidative potency in fruit peel, flesh, and seeds of Saskatoon berry

Food Chemistry 305 (2020) 125430

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Distribution of phytochemicals and antioxidative potency in fruit peel, flesh, and seeds of Saskatoon berry

T

Sabina Lachowicza, , Łukasz Seligab, Stanisław Plutab ⁎

a b

Department of Fermentation and Cereals Technology, Wrocław University of Environmental and Life Science, Chełmońskiego 37 Street, 51-630 Wroclaw, Poland Research Institute of Horticulture, Department of Horticultural Crop Breeding, Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland

ARTICLE INFO

ABSTRACT

Keywords: Cultivars Amelanchier alnifolia Nutt. Polyphenol compounds Tetraterpenoids Triterpenoids Components fruits PCA

The distributions of total soluble solids, pectins, the sum of polyphenolic and terpenoid compounds as well as the antioxidative potency of fruit and their parts, such as peel, flesh, and seeds of Saskatoon berry genotypes are presented in this work. The contents of individual bioactive compounds of in this fruit fraction of Saskatoon berry cultivars and cultivation clones significantly depended on berry genotype and compounds distribution within particular parts of the fruit. The fruit peel contained mainly anthocyanins, polymerized compounds, hydroxycinnamic acids, triterpenoids, and tetraterpenids, as well as exhibited antioxidant activity. The major compounds identified in the fruit flesh included soluble solids, ash, flavonols. In turn, polymeric procyanidins was determined in the seeds of fruit. Individual parts of Saskatoon berry fruit contained many health-promoting constituents and could be deemed attractive materials for the production of functional foods or dietary supplements.

1. Introduction The Saskatoon berry belongs to the Rosaceae family, the Pomoide subfamily. The Latin name for this berry is Amelanchier alnifolia Nutt. This plant is native to the North America, Europe, Africa, and the eastern part of Asia (Lavola, Karjalainen, & Julkunen-Tiitto, 2012; Ozga, Saeed, Wismer, & Reinecke, 2007). The largest area of its cultivation is in Canada. In Europe, it has recently been grown on a very small scale in Finland, Czech Republic. Lithuania, Latvia, and also in Poland. It is often confused because of its similarity to chokeberry (Aronia melanocarpa) and highbush blueberry (Vaccinium corymbosum) and is incorrectly called “sweet chokeberry” or “Canadian blueberry” (Mazza, 2005; Ozga et al., 2007). Fruit of the Saskatoon berry and their parts represent a compendium of health-promoting nutrients and may be used as components of functional foods. They are particularly rich in insoluble and soluble fiber, vitamins (tocopherol, riboflavin, ascorbic acid, pyridoxine, thiamin, riboflavin), minerals (manganese, magnesium, iron, calcium, potassium), sugars (sucrose, glucose, fructose, sorbitol), organic acids, protein, and pectin. 100 g of these fruit provide around 85 kcal of food energy (Lavola et al., 2012; Meda, Mitra, Lee, & Chang, 2016). The chemical composition of Saskatoon berry fruit varies depending on the genotype, maturity at harvest, growing conditions, storage conditions,

and climate (Eisele & Drake, 2005). Saskatoon berries are a great source of phytochemicals, mainly polyphenols, which display strong antioxidant and anti-inflammatory properties and health-promoting benefits (Jin et al., 2015). These compounds have been detected in all anatomical and morphological parts of this plant – skin, flesh, seeds, roots, rhizomes, stems, leaves, and flowers – although they have been not equally distributed. They are also claimed to be responsible for its sensory profile (Lachowicz, Oszmiański, & Pluta, 2017; Lachowicz, Oszmiański, Seliga, & Pluta, 2017). The skin of the Saskatoon berry is rich in anthocyanins (including cyanidin derivatives), flavonols (including quercetin derivatives), and flavanols (including mainly polymeric procyanidins). The peel and skin of the fruit are rich in phenolic acids, including chlorogenic and neochlorogenic acid. Other health properties of the Saskatoon berry are related to the content of carotenoids and triterpenoids which exhibit anticarcinogenic, anti-inflammatory, and antioxidative properties (Lachowicz, Oszmiański, & Pluta, 2017; Lachowicz, Oszmiański, Seliga, & Pluta, 2017). Terpenoids as triterpenoids are distributed mainly in cuticular waxes (Lavola et al., 2012; Loza-Mejía & Salazar, 2015; Ozga et al., 2007). Carotenoids are responsible for the natural pigment of Saskatoon berry fruit skin (Mazza, 2005). Therefore, the main aims of this study were to determine the distribution of bioactive components (triterpenoids, phenolic compounds,

⁎ Corresponding author at: Wrocław University of Environmental and Life Sciences, Faculty of Food Science, Department of Fermentation and Cereals Technology, 37 Chełmońskiego Street, 51-630 Wrocław, Poland. E-mail addresses: [email protected] (S. Lachowicz), [email protected] (Ł. Seliga), [email protected] (S. Pluta).

https://doi.org/10.1016/j.foodchem.2019.125430 Received 1 April 2019; Received in revised form 16 August 2019; Accepted 27 August 2019 Available online 30 August 2019 0308-8146/ © 2019 Elsevier Ltd. All rights reserved.

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tetraterpenoids - chlorophylls and carotenoids), and antioxidative activities of compounds found in different plant parts (fruit, seeds, flesh and peel) in 7 Saskatoon berry genotypes cultivated in Poland. Fruit peel, flesh, and seeds of 4 cultivars (‘Smoky’, ‘Thiessen’, ‘Martin’, ‘Pembina’) and 3 breeding clones ('no 5/6', 'type N' and 'type S') were determined for contents of terpenoid, and polyphenolics using the UPLC-PDA-Q/Tof-MS method. It is expected that this work would allow determining the disparity in the distribution of phytochemicals and identifying the parts with the highest contents of these compounds to enable development of novel functional products.

was ± 1.000 for the mass window. The mass spectrometer was operated in negative-ion mode, set to the base peak intensity (BPI) chromatograms, and scaled to 12,400 counts per second (cps) (100%). The optimized MS conditions were as follows: capillary voltage of 2500 V, cone voltage of 30 V, source temperature of 100 °C, desolvation temperature of 300 °C, and desolvation gas (nitrogen) flow rate of 300 L/h. Collision-induced fragmentation experiments were performed using argon as the collision gas, with voltage ramping cycles from 0.3 to 2 V. The data obtained from UPLC−MS were subsequently entered into the MassLynx 4.0 ChromaLynx Application Manager software. For the extraction and determination of phenolic compounds, a protocol described before by Lachowicz, Oszmiański, and Pluta (2017) and Lachowicz, Oszmiański, Seliga, and Pluta (2017) was followed. The runs were monitored at the following wavelengths: phenolic acids at 320 nm, flavonols at 360 nm, anthocyanins at 520 nm, flavan-3-ols at 280 nm. The PDA spectra were measured over the wavelength range of 200–600 nm in steps of 2 nm. All data were obtained in triplicate. The results were expressed as mg/100 g of dry matter (d.m.).

2. Materials and methods 2.1. Chemicals Acetonitrile, formic acid, methanol, all-trans-β-carotene, α-carotene, all-trans-lutein, neoxanthin, zeaxanthin, pheophorbide, pheophytin a and b, β-cryptoxanthin, chlorophyll b, betulinic, oleanolic and ursolic acid, 2,2-Diphenyl-1-picrylhydrazy, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), methanol, acetic acid, and phloroglucinol were purchased from Sigma-Aldrich (Steinheim, Germany). (−)-Epicatechin, (+)-catechin, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, dicaffeic acid, procyanidin A2, procyanidin B2, p-coumaric acid, caffeic acid, quercetin-3-O-rutinoside, quercetin-3-O-galactoside, quercetin-3O-glucoside, cyanidin-3-O-galactoside and cyanidin-3-O-glucoside were purchased from Extrasynthese (Lyon, France). Acetonitrile for ultraperformance liquid chromatography (UPLC; Gradient grade) and ascorbic acid were from Merck (Darmstadt, Germany).

2.5. Determination of proanthocyanidins Phloroglucinolysis of powder samples was performed according to the protocol described by Lachowicz, Oszmiański, and Pluta (2017) and Lachowicz, Oszmiański, Seliga, and Pluta (2017). Phloroglucinolysis products were separated on a Cadenza CD C18 (75 mm × 4.6 mm, 3 μm) column (Imtakt, Japan). Analysis was carried out using a Waters (Milford, MA) system equipped with Waters 474 diode array and scanning fluorescence detectors and Waters 717 plus autosampler. The mobile solvents were 0.25% aqueous acetic acid (A) and acetonitrile (B). The fluorescence detection was monitored at 278 nm and 360 nm. The calibration curves were established using (+)-catechin and (−)-epicatechin-phloroglucinol adducts standards. All data were obtained in triplicate. The results were expressed as mg/100 g d.m.

2.2. Materials Fruit materials of the Saskatoon berry contain ‘Martin’, ‘Pembina’, ‘Smoky’ and ‘Thiessen’ (Canadian cultivars (cvs.)) and Polish clones as 'no. 5/6', 'type S' and 'type N') and theirs components (peel, flesh and seeds) were tested in the work. Fruit materials were collected at the optimum ripening time. Experimental Orchard at Dąbrowice, belonging to the Research Institute of Horticulture in Skierniewice, Central Poland (51° 55′ 24″ N, 020° 5′ 58″ E). The raw material was directly frozen in liquid nitrogen and freeze-dried (24 h; Christ Alpha 1–4 LSC; Germany). The homogeneous dry material was obtained by crushing the dried tissues using a closed laboratory mill (IKA A.11, Germany). The powders were kept in a refrigerator (−80 °C) until extract preparation.

2.6. Determination of triterpenoids

The fruit soluble solids' amount was analyzed by a refractometer (°Brix), while the amount of pectins was determined according to the Morris method (Pijanowski, Mrożewski, Horubała, & Jarczyk, 1973) (g/ 100 g). Total value of ash was determined by PN-90/A-75101/08.

Fruit sample extraction was performed as described by Lachowicz, Oszmiański, and Pluta (2017), Lachowicz, Oszmiański, Seliga, and Pluta (2017) and Farneti et al. (2015). Identification and quantification of ursolic, oleanolic, and betulinic acids was done using the ACQUITY Ultra Performance LC system with a binary solvent manager (Waters Corp., Milford, MA, USA), a UPLC BEH C18 column (1.7 μm, 2.1 mm × 150 mm, Waters Corp., Milford, MA, USA), and a Q-TOF mass spectrometer (Waters, Manchester, UK) equipped with an electrospray ionization (ESI) source, operating in negative mode. The elution solvent was methanol-acetonitrile (15:85, v/v), at a flow rate of 0.1 mL min−1. The m/z for betulinic acid was 455.3452, for oleanolic acid 455.3496, and for ursolic acid 455.3365, respectively. The compounds were monitored at 210 nm. All data were obtained in triplicate. The results were expressed as mg/kg d.m.

2.4. Determination of polyphenols

2.7. Determination of tetraterpenoids

All analyses of polyphenols of the Saskatoon fruit were carried out using an ACQUITY Ultra Performance LC system (UPLC) equipped with binary solvent manager (Waters Corp., Milford, MA, USA), a UPLC BEH C18 column (1.7 μm, 2.1 mm × 50 mm, Waters Corp., Milford, MA, USA) and a Q-Tof Micro mass spectrometer (Waters, Manchester, UK) with an ESI source operating in negative and positive modes. The analysis was carried out using full-scan, data-dependent MS scanning from m/z 100 to 1500. Leucine enkephalin was used as the reference compound at a concentration of 500 pg/μL, and a flow rate of 2 μL/min, and the [M–H]− ion at 554.2615 Da was detected. The [M–H]− ion was detected during 15 min analysis performed within ESI–MS accurate mass experiments, which were permanently introduced via the LockSpray channel using a Hamilton pump. The lock mass correction

For the extraction of carotenoids, a protocol similar to that described previously was applied (Lachowicz, Oszmiański, & Pluta, 2017; Lachowicz, Oszmiański, Seliga, & Pluta, 2017; Lin & Chen, 2003). Compounds were separated with the ACQUITY UPLC BEH RP C18 column (1.7 μm, 2.1 mm × 100 mm, Waters Corp., Milford, MA, USA) at 32 °C. The elution solvents were ACN:MeOH (7:3, v/v) (A) and 0.1% formic acid (B). Samples (10 μL) were eluted according to the linear gradient described by Delpino-Rius et al. (2014). The runs were monitored at 450 and 650 nm. The PDA spectra were measured over the wavelength range of 200–700 nm in steps of 2 nm. The retention times and spectra were compared to those of the authentic standards. All incubations were done in triplicate. The results were expressed as mg/ kg d.m.

2.3. Physicochemical analysis

2

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probably due to various weather and soil conditions in both countries. Pectins content of the tested berries differed significantly depending on the analyzed part of fruit and genotype. The highest amount of these compounds was found in fruit of 'clone no 5/6' and the lowest one in cv. ‘Thiessen’. The results was similar with the other author (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). Among the analyzed parts, the average content of pectins in the peel was 2.0 times higher compared to the flesh. Pectins were not identified in seeds of the tested berry genotypes (Table 1). Mineral composition of fruits depended on their cultivar, fruit parts, as well as growing conditions including climatic and soil conditions (Lavola et al., 2012). In our study, the average content of ash in fruit of the tested genotypes was 0.5% and ranged from 0.2% in 'clone type N' to 0.7% in cvs. ‘Thiessen’ and ‘Smoky’ (Table 1). The result was similar to that reported earlier by Lachowicz, Oszmiański, Seliga, and Pluta (2017). Among the tested fruit parts, the flesh has the highest mean content of ash, which was 1.4 and 5.4 times higher than in to the peel and seeds. The average content of ash reported previously by Mazza (2005) in the Saskatoon berry genotypes grown in Canada was 0.6% and it was similar to our results.

Table 1 Soluble solid, pectin and ash contents of different fruit parts of seven Saskatoon berry genotypes. Genotypes

Components part

Soluble solids (oBrix)

Pectins (g/100 g)

Ash (%)

Thiessen

Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds

20.3 ± 0.2⁎ 17.9 ± 0.1 12.7 ± 0.1 6.1 ± 0.0 22.5 ± 0.2 175 ± 0.1 14.2 ± 0.1 5.38 ± 0.0 23.2 ± 0.2 25.7 ± 0.1 17.5 ± 0.2 3.7 ± 0.0 14.9 ± 0.1 16.5 ± 0.1 11.2 ± 0.1 1.8 ± 0.0 21.6 ± 0.2 23.0 ± 0.1 13.9 ± 0.2 3.9 ± 0.0 18.5 ± 0.1 20.8 ± 0.1 14.4 ± 0.1 2.5 ± 0.0 17.7 ± 0.1 13.9 ± 0.1 12.1 ± 0.1 4.9 ± 0.0 19.8a 19.4ab 13.7c 4.0d

0.5 ± 0.7 ± 0.5 ± 0.0 ± 0.7 ± 0.9 ± 0.8 ± 0.0 ± 0.6 ± 0.8 ± 0.6 ± 0.0 ± 0.7 ± 0.9 ± 0.6 ± 0.0 ± 0.9 ± 0.9 ± 0.6 ± 0.0 ± 0.7 ± 0.8 ± 0.6 ± 0.0 ± 0.8 ± 0.9 ± 0.7 ± 0.0 ± 0.7a 0.3c 0.6b 0.0d

0.7 ± 0.6 ± 0.4 ± 0.2 ± 0.7 ± 0.5 ± 0.4 ± 0.2 ± 0.5 ± 0.5 ± 0.4 ± 0.1 ± 0.5 ± 0.6 ± 0.4 ± 0.1 ± 0.6 ± 0.6 ± 0.4 ± 0.1 ± 0.6 ± 0.7 ± 0.5 ± 0.1 ± 0.2 ± 0.5 ± 0.4 ± 0.1 ± 0.5ab 0.6a 0.4c 0.1d

Smoky

Martin

Pembina

Clone no 5/6

Clone type S

Clone type N

MEAN of chemical components

0.1 0.1 0.1 0.0 0.1 0.2 0.2 0.0 0.1 0.2 0.1 0.0 0.1 0.2 0.1 0.0 0.2 0.2 0.1 0.0 0.1 0.2 0.1 0.0 0.2 0.2 0.1 0.0

0.1 0.1 0.1 0.0 0.1 0.1 0.1 0.0 0.1 0.1 0.1 0.0 0.1 0.1 0.1 0.0 0.1 0.1 0.1 0.0 0.1 0.1 0.1 0.0 0.0 0.1 0.1 0.0

3.2. Phenolic compounds Contents of anthocyanins, flavonols, flavanols, and phenolic acids in fruit of the investigated Saskatoon berry genotypes and their parts (flesh, peel, and seeds) are shown in Table 2. The mean total content of polyphenols in berries of 7 tested genotypes was 3073 mg/100 g dry matter (d.m.). with the highest contents determined in cultivars: ‘Thiessen’ and ‘Smoky’, and the lowest ones in genotype 'no 5/6′ and cv. ‘Pembina'. Contents of polyphenolics in berry cultivars grown in Poland (‘Thiessen’, ‘Smoky’, ‘Pembina’ and ‘Martin’) were similar to the results obtained previously for the same cultivars grown in Canada and Finland (Bakowska-Barczak & Kolodziejczyk, 2008; Lavola et al., 2012). In addition, they were consistent with findings from our previous research (Lachowicz, Oszmiański, & Pluta, 2017; Lachowicz, Oszmiański, Seliga, & Pluta, 2017; Lachowicz, Oszmiański, Wiśniewski, Seliga, & Pluta, 2019). The mean content of polyphenolics in fruit parts ranged from 3740 mg/100 g d.m. in the peel to 1385 mg/100 g d.m. in the flesh, regardless of the genotype. The results was similar with the others authors (Lachowicz et al., 2019). The peel contained from 44% ('clone no 5/6') to 60% (cv. ‘Thiessen’) of total phenolics, whereas the flesh contained from 16% ('clone type N') to 29% (cv. ‘Martin’), and the seeds from 17% (cv. ‘Smoky’) to 40% (cv. ‘Pembina’) of these compounds. According to Inglett and Chen (2011), the average content of polyphenols in peel, flesh, and seeds of miracle berry (Synsepalum dulcificum) was 1.2, 1.5, and 9.0 times higher than in the same fruit parts of Saskatoon berry. Furthermore, the average content of polyphenols in whole fruit, peel, and flesh of blueberry cv. ‘Bluegold’ was 1.2, 1.5, and 2.7 times lower than in the same fruit parts of the Saskatoon berry genotypes (Ribera, Reyes-Diaz, Alberdi, Zuñiga, & Mora, 2010). These differences were due to the tested species, climate, genotypes, fruit parts analyzed, environmental conditions, and geographical origin (Lavola et al., 2012).

a–d Means - SD followed by different letters within the same line represent significant differences (p < 0.05). ⁎ Values are means ± standard deviation. n = 3.

2.8. Determination of antioxidant activity Antioxidant activity measured by FRAP (Benzie & Strain, 1996) and DPPH (Yen & Chen, 1995) involved UV-2401 PC spectrophotometer (Shimadzu, Kyoto, Japan). After 10 min of reaction, the absorbance was measured at 517 nm for DPPH, and 593 nm for FRAP. The antioxidant potency was expressed as mmol Trolox (TE)/100 g d.m. 2.9. Statistical analysis Statistical analysis such as principal component analysis (PCA) and one-way ANOVA were analyzed using Statistica 12.5 (StatSoft, Kraków, Poland). Significant differences (p < 0.05) between mean values were evaluated by one-way ANOVA and Duncan's multiple range test. 3. Results and discussion

3.2.1. Flavanols The predominating group of polyphenols in fruit of the tested Saskatoon berry and in particular fruit parts was flavanols which accounted for 59% on average, and were followed by the polymeric procyanidins which accounted for 53% on average of total polyphenols. The polymeric procyanidins, like anthocyanins, occurred mainly in fruit peel. Likewise phenolic acids, the monomeric and oligomeric flavonols undergo oxidation under the influence of enzymatic reactions. Their amounts fluctuated between 216 mg/100 g d.m. in cv. ‘Pembina’ and 1434 mg/100 g d.m. in cv. ‘Thiessen’. Among the analyzed berry parts of the tested genotypes, the highest mean content of polymerized compounds was determined in seeds and peel and it was over twice

3.1. Major chemical compounds Contents of total soluble solids determined in fruits of the tested Saskatoon berry and also in their peel, flesh, and seeds are provided in Table 1. The content of soluble solids in fruit of seven tested genotypes ranged from 17.7°Brix in clone type N to 23.2°Brix in cv. ‘Martin’. The highest average content of soluble solids in the tested parts of berries was noted in the flesh (19.4°Brix) and was 1.4 and 4.8 times higher than that determined in the peel and seeds, respectively. According to Mazza (2005), the content of these components in fruit of Saskatoon berry cultivars tested in Canada was 1.7 times lower in comparison with our results obtained for seven genotypes grown in Poland. This was 3

4

Anthocyanins

Compounds

PC

Flavonols

Phenolic acid

Flavanols

Anthocyanins

Compounds

C3E C3P C3H C3G C3Gl C3A C3X

C3E C3P C3H C3G C3Gl C3A C3X PA2 PB2 C(+) E(−) PB3 PB4 PB4 PA3 PB4 PB3 PP DP PrA TQA NA CAG CCA CA 4CA 3PA 2CQA KG Q3A Q3R Q3Rob Q3G Q3Glu Q3Ar Q3Xy Q3AG Q3AGal QDH

1.8 ± 0.0 1.5 ± 0.0 2.0 ± 0.0 515.1 ± 1.0 79.1 ± 0.2 56.0 ± 0.1 60.2 ± 0.1

0.2 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 19.4 ± 0.0 9.0 ± 0.0 6.0 ± 0.0 5.8 ± 0.0

3.0 ± 0.0 1.7 ± 0.0 2.5 ± 0.0 677.0 ± 1.4 120.9 ± 0.2 82.1 ± 0.2 81.4 ± 0.2

0.0 0.0 0.0 1.2 0.4 0.2 0.3

± ± ± ± ± ± ±

Seeds 0.0 0.0 0.0 0.0 0.0 0.0 0.0

0.8 ± 0.0 0.5 ± 0.0 1.0 ± 0.0 481.8 ± 1.0 29.7 ± 0.1 28.5 ± 0.1 29.5 ± 0.1

Fruit

Peel

Fruit

Flesh

Pembina

Martin

0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 15.0 ± 0.0 5.8 ± 0.0 4.1 ± 0.0 4.1 ± 0.0 0.6 ± 0.0 21.1 ± 0.0 16.7 ± 0.0 4.6 ± 0.0 12.3 ± 0.0 1.6 ± 0.0 1.5 ± 0.0 5.3 ± 0.0 0.3 ± 0.0 0.7 ± 0.0 1285.3 ± 2.6 5.3 1.1 ± 0.0 10.4 ± 0.0 9.7 ± 0.0 5.0 ± 0.0 6.9 ± 0.0 51.8 ± 0.1 0.6 ± 0.0 3.3 ± 0.0 1.9 ± 0.0 0.0 ± 0.0 1.4 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 5.6 ± 0.0 0.6 ± 0.0 0.2 ± 0.0 0.3 ± 0.0 0.5 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 1479.9

Flesh 0.0 0.0 0.0 3.4 1.4 1.2 1.2

± ± ± ± ± ± ±

0.0 0.0 0.0 0.0 0.0 0.0 0.0

6.0 ± 0.0 4.0 ± 0.0 5.6 ± 0.0 921.1 ± 1.8 215.4 ± 0.4 152.2 ± 0.3 157.8 ± 0.3 35.6 ± 0.1 92.5 ± 0.2 165.7 ± 0.3 73.1 ± 0.1 82.2 ± 0.2 32.9 ± 0.1 60.4 ± 0.1 66.9 ± 0.1 10.9 ± 0.0 27.5 ± 0.1 816.5 ± 1.6 4.8 52.9 ± 0.1 3.0 ± 0.0 129.5 ± 0.3 37.9 ± 0.1 69.6 ± 0.1 413.8 ± 0.8 5.2 ± 0.0 33.7 ± 0.1 0.6 ± 0.0 1.6 ± 0.0 104.3 ± 0.2 10.7 ± 0.0 7.8 ± 0.0 294.6 ± 0.6 62.9 ± 0.1 8.4 ± 0.0 28.3 ± 0.1 52.3 ± 0.1 5.7 ± 0.0 6.6 ± 0.0 4255.9

1.3 ± 0.0 0.6 ± 0.0 1.0 ± 0.0 104.9 ± 0.2 46.4 ± 0.1 27.7 ± 0.1 28.7 ± 0.1 2.2 ± 0.0 100.9 ± 0.2 109.6 ± 0.2 70.4 ± 0.1 48.6 ± 0.1 9.1 ± 0.0 11.5 ± 0.0 29.9 ± 0.1 2.1 ± 0.0 5.2 ± 0.0 1088.7 ± 2.2 6.7 27.7 ± 0.1 1.6 ± 0.0 106.4 ± 0.2 38.1 ± 0.1 67.1 ± 0.1 303 ± 0.6 26.9 ± 0.1 12.8 ± 0.0 0.3 ± 0.0 4.8 ± 0.0 28.5 ± 0.1 4.8 ± 0.0 2.4 ± 0.0 89.2 ± 0.2 23.8 ± 0.0 15.6 ± 0.0 12.1 ± 0.0 51.3 ± 0.1 6.5 ± 0.0 9.0 ± 0.0 2495.8

6.3 ± 0.0a 3.4 ± 0.0 5.8 ± 0.0 1101.5 ± 2.2 225.1 ± 0.5 157.7 ± 0.3 163.5 ± 0.3 13.6 ± 0.0 106.6 ± 0.2 152.8 ± 0.3 96.2 ± 0.2 65.9 ± 0.1 23.5 ± 0.0 20.7 ± 0.0 41.0 ± 0.1 3.7 ± 0.0 9.4 ± 0.0 1434.4 ± 2.9 5.8 48.6 ± 0.1 4.7 ± 0.0 154.4 ± 0.3 47.3 ± 0.1 117.5 ± 0.2 742.7 ± 1.5 6.4 ± 0.0 46.5 ± 0.1 0.9 ± 0.0 0.4 ± 0.0 38.3 ± 0.1 3.4 ± 0.0 2.6 ± 0.0 228.5 ± 0.5 24.3 ± 0.0 0.9 ± 0.0 9.7 ± 0.0 17.3 ± 0 1.9 ± 0.0 2.2 ± 0.0 5130.5 6.1 ± 0.0 3.0 ± 0.0 5.2 ± 0.0 1280.9 ± 2.6 256.1 ± 0.5 259.5 ± 0.5 272.7 ± 0.5 67.4 ± 0.1 92.7 ± 0.2 149.0 ± 0.3 11.7 ± 0.0 55.9 ± 0.1 50.5 ± 0.1 59.7 ± 0.1 48.1 ± 0.1 28.7 ± 0.1 72.8 ± 0.1 1324.1 ± 2.6 9.6 348.0 ± 0.7 8.9 ± 0.0 178.9 ± 0.4 10.5 ± 0.0 228.6 ± 0.5 845.5 ± 1.7 1.9 ± 0.0 192.7 ± 0.4 1.6 ± 0.0 0.5 ± 0.0 6.2 ± 0.0 1.3 ± 0.0 0.4 ± 0.0 27.4 ± 0.1 4.7 ± 0.0 1.4 ± 0.0 1.5 ± 0.0 2.4 ± 0.0 0.3 ± 0.0 0.3 ± 0.0 5932.9

Fruit

Seeds

Flesh

Fruit

Peel

Smoky

Thiessen

Table 2 The content of polyphenol in seven Saskatoon berry genotypes and their parts [mg/100 g d.m.]

1.2 ± 0.0 0.6 ± 0.0 1.4 ± 0.0 413.3 ± 0.8 41.3 ± 0.1 38.4 ± 0.1 39.9 ± 0.1

Peel

0.0 0.0 0.0 0.7 0.3 0.3 0.3

± ± ± ± ± ± ±

Seeds

0.6 ± 0.0 0.3 ± 0.0 0.5 ± 0.0 50.7 ± 0.1 22.9 ± 0.0 14.5 ± 0.0 15.0 ± 0.0 8.5 ± 0.0 30.8 ± 0.1 23.7 ± 0.0 59.8 ± 0.1 33.3 ± 0.1 16.1 ± 0.0 11.7 ± 0.0 16.8 ± 0.0 2.1 ± 0.0 5.3 ± 0.0 925.9 ± 1.9 5.7 1.2 ± 0.0 0.4 ± 0.0 46.9 ± 0.1 15.7 ± 0.0 13.1 ± 0.0 137.6 ± 0.3 5.7 ± 0.0 33.6 ± 0.1 0.1 ± 0.0 2.2 ± 0.0 111.3 ± 0.2 27.6 ± 0.1 7.6 ± 0.0 227.9 ± 0.5 4.9 ± 0.0 1.3 ± 0.0 24.8 ± 0.0 51.1 ± 0.1 5.6 ± 0.0 6.4 ± 0.0 1963.7

Flesh

0.0 0.0 0.0 0.0 0.0 0.0 0.0

0.0 0.0 0.0 4.5 2.3 1.0 1.6

± ± ± ± ± ± ±

Flesh 0.0 0.0 0.0 0.0 0.0 0.0 0.0

0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3.2 ± 0.0 1.2 ± 0.0 0.9 ± 0.0 0.9 ± 0.0 0.9 ± 0.0 4.5 ± 0.0 1.9 ± 0.0 3.9 ± 0.0 1.4 ± 0.0 0.9 ± 0.0 1.2 ± 0.0 1.3 ± 0.0 0.2 ± 0.0 0.5 ± 0.0 1289.4 ± 2.6 5.6 1.0 ± 0.0 2.1 ± 0.0 2.3 ± 0.0 1.5 ± 0.0 0.9 ± 0.0 11.3 ± 0.0 0.6 ± 0.0 1.6 ± 0.0 0.4 ± 0.0 0.3 ± 0.0 1.1 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 4.6 ± 0.0 0.5 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 0.4 ± 0.0 0.0 ± 0.0 0.1 ± 0.0 1341.9

Seeds

(continued on next page)

0.5 ± 0.0 0.3 ± 0.0 0.6 ± 0.0 536.9 ± 1.1 23.7 ± 0.0 13.9 ± 0.0 12.8 ± 0.0

Fruit

Clone no 5/6

4.5 ± 0.0 2.3 ± 0.0 4.3 ± 0.0 411.8 ± 0.8 1062.3 ± 2.1 116.8 ± 0.2 121.0 ± 0.2 20.7 ± 0.0 47.6 ± 0.1 156.0 ± 0.3 109.1 ± 0.2 57.8 ± 0.1 18.8 ± 0.0 26.1 ± 0.1 35.8 ± 0.1 4.7 ± 0.0 11.9 ± 0.0 1846.4 ± 3.7 10.7 4.9 ± 0.0 1.2 ± 0.0 72.0 ± 0.1 7.8 ± 0.0 46.7 ± 0.1 344.5 ± 0.7 4.6 ± 0.0 31.1 ± 0.1 0.2 ± 0.0 0.3 ± 0.0 9.6 ± 0.0 1.6 ± 0.0 0.2 ± 0.0 40.3 ± 0.1 4.6 ± 0.0 0.2 ± 0.0 2.0 ± 0.0 1.3 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 4631.2

Peel

S. Lachowicz, et al.

Food Chemistry 305 (2020) 125430

5

Anthocyanins

Compounds

PC

Flavonols

Phenolic acid

Flavanols

Compounds

C3E C3P C3H C3G C3Gl C3A C3X

PA2 PB2 C(+) E(−) PB3 PB4 PB4 PA3 PB4 PB3 PP DP PrA TQA NA CAG CCA CA 4CA 3PA 2CQA KG Q3A Q3R Q3Rob Q3G Q3Glu Q3Ar Q3Xy Q3AG Q3AGal QDH

Table 2 (continued)

Seeds

1.8 ± 0.0 0.5 ± 0.0 0.8 ± 0.0 496.0 ± 1 52.5 ± 0.1 26.2 ± 0.1 27.9 ± 0.1

0.0 0.0 0.0 0.3 0.2 0.1 0.1

± ± ± ± ± ± ±

0.0 0.0 0.0 0.0 0.0 0.0 0.0

2.2 ± 0.0 0.6 ± 0.0 1.7 ± 0.0 1109.1 ± 2.2 78.6 ± 0.2 46.2 ± 0.1 47.9 ± 0.1

Fruit

Seeds

Peel

24.7 ± 0.1 90.5 ± 0.2 80.1 ± 0.2 115.9 ± 0.2 99.2 ± 0.2 38.2 ± 0.1 9.2 ± 0.0 48.8 ± 0.1 1.5 ± 0.0 4.2 ± 0.0 1505.9 ± 3.0 10.4 5.9 ± 0.0 2.1 ± 0.0 153.9 ± 0.3 25.3 ± 0.0 4.0 ± 0.0 384.9 ± 0.8 1.4 ± 0.0 15.0 ± 0.0 0.4 ± 0.0 0.3 ± 0.0 4.9 ± 0.0 0.7 ± 0.0 0.0 ± 0.0 13.0 ± 0.0 2.2 ± 0.0 0.4 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3601.5 Clone type S

6.0 ± 0.0 3.8 ± 0.0 17.4 ± 0.0 0.0 ± 0.0 15.8 ± 0.0 3.3 ± 0.0 4.3 ± 0.0 4.5 ± 0.0 1.0 ± 0.0 1.9 ± 0.0 929.0 ± 1.9 7.6 1.1 ± 0.0 2.6 ± 0.0 25.7 ± 0.1 2.9 ± 0.0 0.4 ± 0.0 64.3 ± 0.1 65.0 ± 0.1 1.7 ± 0.0 0.3 ± 0.0 0.8 ± 0.0 106.6 ± 0.2 8.9 ± 0.0 1.0 ± 0.0 409.3 ± 0.8 49.2 ± 0.1 10.0 ± 0.0 2.5 ± 0.0 8.0 ± 0.0 0.8 ± 0.0 1.2 ± 0.0 1790.0

Clone no 5/6

24.1 ± 0.0 62.9 ± 0.1 46.3 ± 0.1 28.1 ± 0.1 44.7 ± 0.1 20.1 ± 0.0 28.9 ± 0.1 23.1 ± 0.0 5.9 ± 0.0 13.0 ± 0.0 800.0 ± 1.6 4.9 7.4 ± 0.0 38.1 ± 0.1 77.9 ± 0.2 13.3 ± 0.0 12.0 ± 0.0 284.1 ± 0.6 3.9 ± 0.0 11.0 ± 0.0 6.0 ± 0.0 0.8 ± 0.0 81.0 ± 0.2 1.0 ± 0.0 3.0 ± 0.0 380.9 ± 0.8 39.2 ± 0.1 4.5 ± 0.0 10.0 ± 0.0 22.0 ± 0.0 2.4 ± 0.0 2.1 ± 0.0 2813.4

0.3 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 17.1 ± 0.0 12.3 ± 0.0 6.8 ± 0.0 7.1 ± 0.0

Flesh

0.0 ± 0.0 1.7 ± 0.0 1.8 ± 0.0 13.4 ± 0.0 0.1 ± 0.0 0.4 ± 0.0 0.5 ± 0.0 1.3 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 1200.2 ± 2.4 7.2 1.0 ± 0.0 0.5 ± 0.0 2.0 ± 0.0 0.5 ± 0.0 0.3 ± 0.0 9.1 ± 0.0 0.4 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 1.0 ± 0.0 0.4 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 3.0 ± 0.0 0.5 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 124.2

3.0 ± 0.0 0.8 ± 0.0 2.2 ± 0.0 1145.1 ± 2.3 107.7 ± 0.2 60.8 ± 0.1 63 ± 0.1

Peel

39.1 ± 0.1 35.3 ± 0.1 51.6 ± 0.1 27.3 ± 0.1 12.4 ± 0.0 8.6 ± 0.0 4.5 ± 0.0 14.2 ± 0.0 0.8 ± 0.0 2.1 ± 0.0 215.9 ± 0.4 4.9 8.7 ± 0.0 7.1 ± 0.0 67.8 ± 0.1 19.9 ± 0.0 123.4 ± 0.2 80.8 ± 0.2 1.4 ± 0.0 12.8 ± 0.0 1.3 ± 0.0 0.7 ± 0.0 30.6 ± 0.1 57.3 ± 0.1 1.0 ± 0.0 10.9 ± 0.0 2.2 ± 0.0 3.2 ± 0.0 1.9 ± 0.0 6.6 ± 0.0 0.7 ± 0.0 0.8 ± 0.0 1423.5

Fruit

Peel

Fruit

Flesh

Pembina

Martin

0.0 0.0 0.0 1.0 0.6 0.7 0.7

± ± ± ± ± ± ±

Seeds 0.0 0.0 0.0 0.0 0.0 0.0 0.0

3.5 ± 0.0 15.3 ± 0.0 14.2 ± 0.0 4.7 ± 0.0 4.3 ± 0.0 4.1 ± 0.0 1.8 ± 0.0 4.3 ± 0.0 0.3 ± 0.0 0.8 ± 0.0 237.7 ± 0.5 4.8 0.3 ± 0.0 0.2 ± 0.0 16.4 ± 0.0 16.1 ± 0.0 3.2 ± 0.0 18.6 ± 0.0 1.5 ± 0.0 39.3 ± 0.1 0.0 ± 0.0 33.6 ± 0.1 49.2 ± 0.1 10.4 ± 0.0 6.7 ± 0.0 94.2 ± 0.2 19.4 ± 0.0 6.7 ± 0.0 12.9 ± 0.0 45.2 ± 0.1 4.9 ± 0.0 5.7 ± 0.0 676.8

Flesh

2.0 ± 0.0 0.9 ± 0.0 1.7 ± 0.0 1158.3 ± 2.3 73.2 ± 0.1 46.7 ± 0.1 48.4 ± 0.1

Fruit

Clone type N

33.8 ± 0.1 38.7 ± 0.1 78.4 ± 0.2 53.6 ± 0.1 33.3 ± 0.1 15.4 ± 0.0 19.5 ± 0.0 23.1 ± 0.0 3.5 ± 0.0 8.9 ± 0.0 1055 ± 2.1 6.0 1.5 ± 0.0 8.7 ± 0.0 51.6 ± 0.1 10.1 ± 0.0 72.5 ± 0.1 111.5 ± 0.2 0.5 ± 0.0 1.2 ± 0.0 1.6 ± 0.0 0.1 ± 0.0 1.1 ± 0.0 2.0 ± 0.0 0.3 ± 0.0 4.7 ± 0.0 1.0 ± 0.0 0.5 ± 0.0 1.5 ± 0.0 2.0 ± 0.0 0.2 ± 0.0 0.2 ± 0.0 2179.0

Peel

0.2 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 12.9 ± 0.0 6.7 ± 0.0 3.7 ± 0.0 3.8 ± 0.0

Flesh

1.0 ± 0.0 0.8 ± 0.0 1.4 ± 0.0 0.4 ± 0.0 0.6 ± 0.0 0.5 ± 0.0 0.4 ± 0.0 0.5 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 1920.9 ± 3.8 16.4 0.0 ± 0.0 0.2 ± 0.0 1.7 ± 0.0 0.3 ± 0.0 4.2 ± 0.0 3.0 ± 0.0 1.2 ± 0.0 0.3 ± 0.0 0.0 ± 0.0 0.5 ± 0.0 0.8 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 1.2 ± 0.0 0.3 ± 0.0 0.1 ± 0.0 0.3 ± 0.0 0.5 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 1943.4

Seeds

0.0 0.0 0.0 2.4 1.0 0.8 0.8

± ± ± ± ± ± ±

Seeds 0.0 0.0 0.0 0.0 0.0 0.0 0.0

2.0 ± 0.0 2.8 ± 0.0 3.5 ± 0.0 8.4 ± 0.0 2.0 ± 0.0 1.1 ± 0.0 1.8 ± 0.0 1.6 ± 0.0 1.0 ± 0.0 3.1 ± 0.0 414.8 ± 0.8 5.6 0.3 ± 0.0 0.3 ± 0.0 15.3 ± 0.0 5.1 ± 0.0 0.5 ± 0.0 37.4 ± 0.1 0.2 ± 0.0 0.3 ± 0.0 0.4 ± 0.0 1.3 ± 0.0 88.4 ± 0.2 12.9 ± 0.0 1.7 ± 0.0 216.1 ± 0.4 16.0 ± 0.0 6.6 ± 0.0 12.9 ± 0.0 10.0 ± 0.0 0.9 ± 0.0 1.0 ± 0.0 877.5

Flesh

(continued on next page)

4.1 ± 0.0 1.8 ± 0.0 3.5 ± 0.0 1310.6 ± 2.6 145.9 ± 0.3 96.7 ± 0.2 100.2 ± 0.2

Peel

10.1 ± 0.0 20.4 ± 0.0 22.1 ± 0.0 18.0 ± 0.0 17.4 ± 0.0 7.8 ± 0.0 8.1 ± 0.0 8.9 ± 0.0 1.6 ± 0.0 3.5 ± 0.0 899.9 ± 1.8 6.6 0.7 ± 0.0 7.0 ± 0.0 47.0 ± 0.1 3.0 ± 0.0 3.0 ± 0.0 136.4 ± 0.3 1.8 ± 0.0 3.5 ± 0.0 0.2 ± 0.0 0.6 ± 0.0 40.2 ± 0.1 5.0 ± 0.0 2.0 ± 0.0 92.3 ± 0.2 5.8 ± 0.0 4.8 ± 0.0 5.6 ± 0.0 17.4 ± 0.0 1.3 ± 0.0 2.4 ± 0.0 1980.2

Fruit

Clone no 5/6

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6

PA2 PB2 C(+) E(−) PB3 PB4 PB4 PA3 PB4 PB3 PP DP PrA TQA NA CAG CCA CA 4CA 3PA 2CQA KG Q3A Q3R Q3Rob Q3G Q3Glu Q3Ar Q3Xy Q3AG Q3AGal QDH

13.0 ± 0.0 30.1 ± 0.1 20.5 ± 0.0 39.4 ± 0.1 7.8 ± 0.0 14.8 ± 0.0 13.9 ± 0.0 9.1 ± 0.0 0.7 ± 0.0 2.8 ± 0.0 955.9 ± 1.9 5.5 8.8 ± 0.0 3.5 ± 0.0 54.1 ± 0.1 7.3 ± 0.0 9.0 ± 0.0 133.2 ± 0.3 0.3 ± 0.0 2.0 ± 0.0 0.5 ± 0.0 0.0 ± 0.0 4.0 ± 0.0 0.6 ± 0.0 4.3 ± 0.0 4.9 ± 0.0 0.6 ± 0.0 0.2 ± 0.0 0.3 ± 0.0 0.8 ± 0.0 1.0 ± 0.0 1.7 ± 0.0 2098.1

2.3 ± 0.0 0.5 ± 0.0 0.7 ± 0.0 1.5 ± 0.0 0.5 ± 0.0 0.6 ± 0.0 0.3 ± 0.0 0.5 ± 0.0 0.6 ± 0.0 2.0 ± 0.0 1454.1 ± 2.9 14.2 0.0 ± 0.0 0.4 ± 0.0 1.0 ± 0.0 0.2 ± 0.0 0.3 ± 0.0 3.7 ± 0.0 0.1 ± 0.0 0.0 ± 0.0 0.3 ± 0.0 0.8 ± 0.0 0.6 ± 0.0 0.1 ± 0.0 4.0 ± 0.0 1.0 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 0.2 ± 0.0 0.3 ± 0.0 0.5 ± 0.0 0.9 ± 0.0 1478.6

2.3 ± 0.0 17.0 ± 0.0 22.0 ± 0.0 20.6 ± 0.0 9.4 ± 0.0 3.5 ± 0.0 4.6 ± 0.0 16.2 ± 0.0 2.0 ± 0.0 1.1 ± 0.0 551.2 ± 1.1 5.9 3.7 ± 0.0 3.5 ± 0.0 34.0 ± 0.1 12.3 ± 0.0 182.4 ± 0.4 101.6 ± 0.2 3.6 ± 0.0 2.0 ± 0.0 0.7 ± 0.0 0.6 ± 0.0 29.5 ± 0.1 5.7 ± 0.0 1.3 ± 0.0 117.8 ± 0.2 11.5 ± 0.0 2.6 ± 0.0 4.9 ± 0.0 3.6 ± 0.0 5.4 ± 0.0 2.5 ± 0.0 2476.0

Fruit

Peel

Seeds

Clone type S

Clone no 5/6

2.0 ± 0.0 1.6 ± 0.0 8.5 ± 0.0 3.3 ± 0.0 1.5 ± 0.0 2.1 ± 0.0 2.2 ± 0.0 6.3 ± 0.0 3.2 ± 0.0 2.1 ± 0.0 1052.5 ± 2.1 4.8 1.6 ± 0.0 9.9 ± 0.0 50.6 ± 0.1 11.4 ± 0.0 212.1 ± 0.4 250.3 ± 0.5 3.2 ± 0.0 4.5 ± 0.0 0.4 ± 0.0 0.5 ± 0.0 27.4 ± 0.1 8.2 ± 0.0 0.8 ± 0.0 113.8 ± 0.2 11.2 ± 0.0 1.8 ± 0.0 3.3 ± 0.0 7.0 ± 0.0 2.2 ± 0.0 0.9 ± 0.0 1859.1

Flesh 9.2 ± 0.0.0 16.9 ± 0.0 40.5 ± 0.1 18.9 ± 0.0 21.2 ± 0.0 10.8 ± 0.0 14.4 ± 0.0 34.5 ± 0.1 5.2 ± 0.0 2.5 ± 0.0 1244.2 ± 2.5 5.4 54.3 ± 0.1 13.0 ± 0.0 108.2 ± 0.2 28.2 ± 0.1 417.7 ± 0.8 378.7 ± 0.8 9.4 ± 0.0 8.6 ± 0.0 0.7 ± 0.0 3.2 ± 0.0 2.6 ± 0.0 0.9 ± 0.0 2.1 ± 0.0 11.2 ± 0.0 1.2 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 0.8 ± 0.0 3.6 ± 0.0 2.1 ± 0.0 3843.4

Peel 0.9 ± 0.0 1.1 ± 0.0 1.0 ± 0.0 0.3 ± 0.0 0.3 ± 0.0 0.2 ± 0.0 0.2 ± 0.0 1.9 ± 0.0 2.6 ± 0.0 1.2 ± 0.0 1480 ± 3 15.0 2.0 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 0.3 ± 0.0 10.6 ± 0.0 10.1 ± 0.0 1.2 ± 0.0 1.0 ± 0.0 0.3 ± 0.0 0.0 ± 0.0 0.6 ± 0.0 0.1 ± 0.0 1.3 ± 0.0 1.6 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 2.4 ± 0.0 1.3 ± 0.0 1528.8

Seeds 6.0 ± 0.0 3.6 ± 0.0 16.6 ± 0.0 4.8 ± 0.0 2.8 ± 0.0 4.4 ± 0.0 8.2 ± 0.0 13.5 ± 0.0 3.7 ± 0.0 1.6 ± 0.0 1255.4 ± 2.5 6.7 95.9 ± 0.2 19.7 ± 0.0 162.2 ± 0.0 2.3 ± 0.7 6.6 ± 0.0 371.4 ± 0.3 2.1 ± 0.0 25.1 ± 0.1 0.6 ± 0.0 0.6 ± 0.0 10.2 ± 0.0 3.5 ± 0.0 1.7 ± 0.0 61.5 ± 0.1 7.1 ± 0.0 1.2 ± 0.0 5.8 ± 0.0 0.3 ± 0.0 3.9 ± 0.0 1.6 ± 0.0 3436.9

Fruit

Clone type N

3.0 ± 0.0 5.4 ± 0.0 6.2 ± 0.0 2.3 ± 0.0 1.3 ± 0.0 1.4 ± 0.0 1.4 ± 0.0 4.5 ± 0.0 2.2 ± 0.0 1.7 ± 0.0 587.7 ± 1.2 5.1 35.9 ± 0.1 4.1 ± 0.0 115.1 ± 0.0 1.2 ± 0.2 1.7 ± 0.0 67.5 ± 0.1 0.4 ± 0.0 3.1 ± 0.0 0.6 ± 0.0 1.1 ± 0.0 22.3 ± 0.0 10.4 ± 0.0 1.6 ± 0.0 142.7 ± 0.3 16.3 ± 0.0 1.7 ± 0.0 7.0 ± 0.0 17.6 ± 0.0 4.1 ± 0.0 2.0 ± 0.0 1102.4

Flesh 2.1 ± 0.0 25 ± 0.0 27.8 ± 0.1 9.5 ± 0.0 5.7 ± 0.0 11.1 ± 0.0 11.9 ± 0.0 18.9 ± 0.0 4.7 ± 0.0 3.0 ± 0.0 1403.3 ± 2.8 6.9 85.8 ± 0.2 11.4 ± 0.0 140.2 ± 0.0 3.3 ± 0.8 9.1 ± 0.0 403.7 ± 0.3 1.1 ± 0.0 41.8 ± 0.1 0.6 ± 0.0 0.1 ± 0.0 1.4 ± 0.0 0.5 ± 0.0 1.5 ± 0.0 7.0 ± 0.0 0.9 ± 0.0 0.2 ± 0.0 0.1 ± 0.0 0.7 ± 0.0 2.5 ± 0.0 1.3 ± 0.0 3900.8

Peel

1.6 ± 0.0 2.0 ± 0.0 4.3 ± 0.0 4.6 ± 0.0 6.2 ± 0.0 1.8 ± 0.0 0.5 ± 0.0 4.0 ± 0.0 3.0 ± 0.0 2.5 ± 0.0 1691.9 ± 3.4 14.7 6.1 ± 0.0 0.8 ± 0.0 0.3 ± 0.0 31.8 ± 0.1 0.9 ± 0.0 11.9 ± 0.0 0.3 ± 0.0 1.1 ± 0.0 0.5 ± 0.0 0.1 ± 0.0 0.7 ± 0.0 0.3 ± 0.0 2.9 ± 0.0 2.7 ± 0.0 0.3 ± 0.0 0.1 ± 0.0 0.1 ± 0.0 0.5 ± 0.0 2.8 ± 0.0 1.3 ± 0.0 1800.3

Seeds

Values are means ± standard deviation. n = 3; Explanations: PC, sum of phenolic compounds; C3E cyanidin-3-hexoside-(epi)catechin; C3P cyanidin-3-pentoside-(epi)catechin; C3H cyanidin-3-hexoside- (epi)cat-(epi) cat; C3G cyanidin-3-O-galactoside; C3Glu cyanidin-3-O-glucoside;C3A cyanidin-3-O-arabinoside; C3X xyanidin-3-O-xyloside; PA2 A-type procyjanidin dimer; PB2 B-type procyjanidin dimer; C(+) (+)-Catechin; PB3 Btype procyjanidin trimer; E(−) (−)-Epicatechin; PB4 B-type procyjanidin tertramer; PA3 A-type procyjanidin trimer; PP Polymeric procyjanidin; DP degree polimeryzation; PrA protocatechuic acid; TQA trihydroxycinnamoylquinic acid isomers; NA neochlorogenic acid; CAG caffeic acid glucoside; CA chlorogenic acid; CCA cryptochlorogenic acid; 3PA 3-O-p-coumaroylquinic acid; 4CA 4-caffequinic acid; 2CQA di-caffeic quinic acid; KG kampferol-3-galactoside; Q3A quercetin-3-O-arabinoglucoside; Q3R quercetin-3-O-rutinoside; Q3Rob quercetin-3-O-robinobioside; Q3G quercetin-3-O-galactoside; Q3Glu quercetin-3-O-glucoside;Q3Ar quercetin3-O-arabinoside; Q3Xy quercetin-3-O-xyloside; G3AG quercetin-3-O-(6″-acetyl)galactoside; Q3AGal quercetin-3-O-(6″-acetyl)galactoside; QDH quercetin-deoxyhexo-heksoside.

a

PC

Flavonols

Phenolic acid

Flavanols

Compounds

Table 2 (continued)

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Food Chemistry 305 (2020) 125430

Food Chemistry 305 (2020) 125430

S. Lachowicz, et al.

higher compared to the flesh. The high content of these compounds in seeds of berry is confirmed by the sour, bitter, and tart flavor. In addition, the highest degree of polymerization was identified in the seeds and it was 2.5 and 2.2 times higher compared to the fruit flesh and peel. These differences could be influenced by the maturation process, exposure to light, cultivar or environmental conditions (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). In addition, contents of monomers and oligomers of flavanols detected in fruit of the tested Saskatoon berry genotypes and their parts (flesh, peel and seeds) reached on average from 65 mg/100 g d.m. in 'clone type S' to 648 mg/100 g d.m. in cv. ‘Smoky’. The highest mean content of these compounds was analyzed in the peel and it was 3 and 13 times higher than in the flesh and seeds. The major monomeric compounds of all studied genotypes and their parts were (+)-catechin, with the mean content at 45 mg/100 g d.m., and also (−)-epicatechin, with the content ranging from 3.27 mg/100 g d.m. in the flesh ('clone type S') to 117 mg/100 g d.m. (cv. ‘Martin’). The highest content of this compound was analyzed in the peel, compared to the other fruit parts of the tested berry genotypes. Another important compounds were procyanidin B2 and B3, with their average contents reaching 31 and 23 mg/100 g d.m., respectively. According to Lavola et al. (2012), the content of flavanols in fruit of Saskatoon berry genotypes cultivated in Finland was slightly (1.2 times) higher than in our study.

differed significantly depending on the analyzed part of berries. Their total content ranged from 202 mg/100 g d.m. ('clone type 5/6') to 1169 mg/100 g d.m. (cv. ‘Thiessen’). These results were in agreement with previously published reports (Jin et al., 2015; Lachowicz, Oszmiański, Seliga, & Pluta, 2017; Lavola et al., 2012; Olthof et al., 2003). The major compound among phenolic acids was chlorogenic acid which accounted for 53% of total hydroxycinnamic acids, with its mean content reaching 358 mg/100 g d.m. on average, regardless of the genotype. Its highest content was found in the peel and it ranged from 11 mg/100 g d.m. (cv. ‘Pembina’) to 845 mg/100 g d.m. (cv. ‘Thiessen’). It was 3 and 23 times higher compared to the flesh and seeds, respectively. Different amounts of cryptochlorogenic and neochlorogenic acids were also noted in various parts of fruit of the studied genotypes and they accounted on average for 14% and 15% in the flesh and peel and for 6% and 7% in seeds, respectively of total hydroxycinnamic acids. In the case of caffeic glucoside and 4-caffequinic acids, they were found mainly in the flesh with an average content of 13 mg/100 g d.m., which was 2 and 12 times higher compared to fruit peel and seeds, respectively. It should be noted that the fruit peel and seeds in the form of processing waste could be valuable sources of anthocyanins as well as polymerized compounds and phenolic acids with health-promoting properties as well as with anti-aging, antioxidative, and anticarcinogenic potency (Farneti et al., 2015). Several authors confirmed chlorogenic, neochlorogenic, and cryptochlorogenic acids to be the major hydroxycinnamic acids in fruits of the Saskatoon berry genotypes (Bakowska-Barczak & Kolodziejczyk, 2008; Farneti et al., 2015; Lachowicz, Oszmiański, & Pluta, 2017; Lachowicz, Oszmiański, Seliga, & Pluta, 2017; Lavola et al., 2012).

3.2.2. Anthocyanins The second group of compounds analyzed in the fruit of seven genotypes of the Saskatoon berry and their peel, flesh, and seeds included seven anthocyanins, identified with the LC/MS technique. Their content in the fruit of the tested genotypes ranged from 572 mg/100 g d.m. in cv. ‘Pembina’ to 1663 mg/100 g d.m. in cv. ‘Thiessen’. The mean anthocyanin content in fruit of the Saskatoon berry cultivars grown in Finland and Alberta (Canada) was 1.3 times higher and 1.2 times lower, than in the fruit of our tested genotypes grown in Poland (BakowskaBarczak & Kolodziejczyk, 2008; Farneti et al., 2015; Lavola et al., 2012). In addition, according to Rockenbach et al. (2011), the content of anthocyanins in red grape (Vitis vinifera) was around 2 times lower than in fruit of the Saskatoon berry. It was confirmed that the content of anthocyanins differed significantly between the analyzed parts of berries. Their higher content was determined in the peel (average 1231 mg/100 g d.m.), regardless of the genotype. The lowest content of anthocyanins was assayed in the seeds and it was around 28 times lower compared to the peel. The major compounds among anthocyanins were cyanidins: -3-O-galactoside and -3-O-glucoside. They accounted for 75% and 20% of total anthocyanins and their average contents in the tested genotypes were at 860 mg/100 g d.m. and 113 mg/100 g d.m. respectively. Their average content in the peel was 791 mg/100 g d.m. and it was 25 times higher than in the flesh of fruit. Seeds contained only traces of anthocyanins. It has been proved that the peel was the richest source of anthocyanins (Oszmiański & Lachowicz, 2016). The content of another anthocyanin, cyanidin-3-O-xyloside, varied also depending on the analyzed fruit parts and accounted on average for 7% of total anthocyanins (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). Various contents of anthocyanins in the tested berry fruits and their parts depended on environmental and climatic factors, cultivar, growing location, and/or exposure to light (Ozga et al., 2007). Furthermore, according to Ribera et al. (2010), the average content of anthocyanins in whole fruit, peel, and flesh of the highbush blueberry (V. corymbosum) cv. ‘Bluegold’ was 1.4, 1.2, and 3.1 times lower than in the peel, flesh, seeds of the berry genotypes tested in our study.

3.2.4. Flavonols Among all polyphenols, detected in the fruit of the analyzed genotypes and in their parts (peel, flesh, seeds), the smallest group (3–16%) included 11 flavonols that were identified with the LC/MS technique (Table 2). Their content ranged from 97 mg/100 g d.m. in 'clone type N' to 583 mg/100 g d.m. in cv. ‘Smoky’. They were found mainly in the flesh of fruit of the analyzed genotypes, where their mean content reached 339 mg/100 g d.m. and accounted for 28% of total polyphenolic compounds in the fruit flesh. About 12 and 40 times lower contents of flavonols were determined in the peel and seeds, respectively, than in the flesh. In the present study, quercetin-3-O-galactoside was the major compound which accounted for 58% of total flavonols on average. Its content ranged from 38% ('clone type N') to 59% (cv ‘Smoky’) in fruit flesh of the analyzed Saskatoon berry genotypes (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). A high content was also determined for quercetin-3-O-arabinoglucoside in fruit and their individual parts. Its mean content was 48 mg/100 g d.m. in berries of the tested genotypes. The highest mean content of this compound was found in the flesh (61 mg/100 g d.m.) and was 14 and 25 times lower than in the peel and seeds, respectively. It means that flavonols accumulated principally in the fruit flesh and only their trace amounts were found in the peel and seeds. According to Inglett and Chen (2011), the average content of flavonols in the peel, flesh, and seeds of miracle berry (Synsepalum dulcificum) was around 1.1, 1.2, and 5.0 times higher than in the tested Saskatoon berry genotypes. As it was presented by Orqueda et al. (2017), the content of flavonols in the whole fruit, peel, flesh, and seeds of the tree tomato or chilto (Argentinean Solanum betaceum) was 2.3 and 1.8 times lower as well as, 2.4 and 10.1 times higher, respectively, compared to the whole fruit and its parts of the Saskatoon berry genotypes tested in this study. The content of polyphenolic compounds differed significantly depending on the analyzed fruit part and genotype. Results from this study showed that fruit of cv. ‘Thiessen’, 'clones type S', and 'type N' and their peel contained mainly anthocyanin, polymerized compounds, and hydroxycinnamic acids. Polymeric procyanidins and phenolic acids were identified in the fruit flesh, while polymerized compounds were

3.2.3. Phenolic acids The phenolic acids were also identified in the fruit of seven Saskatoon berry genotypes and in particular fruit parts using the LC/MS technique (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). The phenolic acids consisted of 10 compounds and accounted on average for 11–27% of total polyphenolics (Table 2). The content of hydroxycinnamic acids 7

Food Chemistry 305 (2020) 125430

S. Lachowicz, et al.

mainly found in the seeds. Fruit of cvs. ‘Smoky’ and ‘Martin’ and their peel contained mainly anthocyanins and polymeric procyanidins (flavanols group). However, flavonols were the major compounds found in the flesh, while polymerized compounds –in the seeds. Fruits of the breeding 'clone no 5/6' and their peel, flesh, and seeds were characterized by high amounts of procyanidin polymers compared to the other identified compounds. In addition, significant amounts of flavonols were also identified in the fruit flesh of this clone. In the case of cv. ‘Pembina’, study results proved a similar distribution of all compounds, while they showed a higher amount of polymeric procyanidins in the peel and seeds, and of flavonols and polymeric procyanidins in the flesh.

higher compared to the flesh and seeds. Among the carotenoids, alltrans-lutein and all-trans-β-carotene were the most commonly noted compounds in fruit parts of different Saskatoon berry genotypes. The peel was the richest source of both compounds and their average contents were 192 mg/kg d.m. and 86 mg/kg d.m., respectively. Contents of all-trans-β-carotene and all-trans-lutein in the peel were 1.6 and 1.8 times higher than in the flesh and seeds, respectively. In turn, Kohno et al. (2001), Razungles, Oszmianski, and Sapis (1989) as well as Mazza and Cottrell (2008) showed that all-trans-β-carotene was the major compound among the identified tetraterpenoids in the tested Saskatoon berry genotypes. Their findings were in an agreement with our results. Furthermore, the average content of carotenoids in the fruit, peel, flesh, and seeds of Argentinean Solanum betaceum was 3.7, 3.3, 3.5, and 5.0 times lower compared to the results of our analyses (Orqueda et al., 2017). In this study, the mean content of chlorophylls determined in seven Saskatoon berry genotypes, and it was similar to that reported earlier by Lachowicz, Oszmiański, Seliga, and Pluta (2017) and Lachowicz et al. (2019) for these genotypes. The highest content of chlorophylls was noted in the peel and it ranged from 70 mg/kg d.m. (cv. ‘Pembina’) to 144 mg/kg d.m. ('clone type S'). For all the tested genotypes, it was on average 2.2 and 3.0 times higher than in the other fruit parts. The most commonly found compound was pheophytin and its content ranged from 23 mg/kg d.m. ('clone no 5/6') to 43 mg/kg d.m. (cv. ‘Thiessen’). Generally, the fruit of the tested Saskatoon genotypes contained a low amount of identified chlorophylls, which indicated the maturity of the raw material under study. The same observation was made Pumilia et al. (2014), who stated that the base content of chlorophylls in the berries indicated their maturity. While, the high content of pheophytin in the peel indicated a greater exposure to temperature when drying the plant material or fruits (Pumilia et al., 2014; DelgadoPelayo, Gallardo-Guerrero, & Hornero-Méndez, 2014).

3.3. Triterpenoid Contents of triterpenoid compounds in the fruit of the investigated Saskatoon berry and their parts are presented in Table 4. Betulinic, oleanolic, and ursolic acids were the three main triterpenoids identified in the whole fruit and their parts using the LC/MS technique (Lachowicz, Oszmiański, & Pluta, 2017; Lachowicz, Oszmiański, Seliga, & Pluta, 2017). The total triterpenoids content in fruit ranged from 31.7 mg (‘Pembina’ cv.) to 71.7 mg ('Thiessen') per kg d.m. The results were 1.3 times lower compared to the Saskatoon berry cultivars and clones from Poland cultivated 2016 and 2017 year (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). The highest average content (92.3 mg/kg d.m.) of triterpenoids was determined in the peel and was 3 and 2 times higher compared to the flesh and seeds, regardless of the genotype. Therefore, it might be concluded that triterpenoid compounds accumulated principally in the peel and seeds, and that only their trace amounts were found in fruit flesh. However, for cv. ‘Smoky’ and 'clones no 5/6' and 'type N', the betulinic acid predominated in the peel compared to the other fruit parts of berry genotypes tested in our study. In the fruit of the analyzed Saskatoon berry genotypes, the major compounds were ursolic acid (40–51%), followed by oleanolic and betulinic acids (27–40% and 17–24%, respectively). Ursolic acid – as the main triterpenoid compound – was also found in cranberry (mean 20% of all wax extract), apple peel (mean 98%), sweet cherry (mean 60%), and pear (mean 50%) (Hussain et al., 2015; Szakiel, Niżyński, & Pączkowski, 2013; Szakiel, Pączkowski, & Huttunen, 2012; Lachowicz, Oszmiański, Seliga, & Pluta, 2017; Sun, Tao, & Zhang, 2019) According to Szakiel, Pączkowski, and Huttunen (2012), contents of oleanolic and ursolic acids found in bilberry fruit cultivated in Finland and Poland were 6.0 and 1.4 times and 7.0 and 1.5 times lower compared to the fruit of Saskatoon berry. Oleanolic and ursolic acids were the major compounds in the bilberry fruit. The average content of ursolic acid in pear was 3 times higher than in the fruit of the tested Saskatoon berry genotypes (Sun et al., 2019). In contrast, Kondo (2006) showed that the average content of triterpenoids in cranberry fruit was twice lower than in the fruit of the genotypes analyzed in our study. These compounds are very important considering their health benefits for the human body. They accumulate mainly in the wax layer of fruit and exhibit antiaging, antioxidative, and anticarcinogenic potency (Neto, 2011).

3.5. Antioxidant potential and correlations The present study included also in vitro tests of the ferric reducing antioxidant capacity (FRAP) and radical scavenging activity (DPPH) that were conducted in the fruit of Saskatoon berry genotypes and also in particular fruit parts (Table 4). Their results confirmed significant differences in contents of antioxidants in the fruit of the tested genotypes and their parts. The antioxidant activity of all Saskatoon berry genotypes was on average 15.11 mmol TE/100 g d.m. to FRAP and 23.9 mmol TE/100 g d.m. to DPPH assays. These results were in an agreement with the previously published data (Lachowicz, Oszmiański, Seliga, & Pluta, 2017). In general, the content of antioxidants determined in Saskatoon berry genotypes was by 40% lower than in chokeberry (Oszmiański & Lachowicz, 2016) and by 20% higher than in cranberry (V. macrocarpon) (Oszmiański, Kolniak-Ostek, Lachowicz, Gorzelany, & Matłok, 2015). The highest antioxidant activity was noted in the peel and ranged from 14.5 mmol TE/100 g (cv. ‘Pembina’) to 41.5 mmol TE/100 g (cv. ‘Thiessen’) when measured with the FRAP method as well as from 32.3 mmol TE/100 g d.m. (clone no 5/6) to 57.9 mmol TE/100 g d.m. (‘clone type N’) when measured with the DPPH assay. The content of antioxidants in the peel was on average 5 and 16 times higher compared to that found in the flesh and seeds. According to a publication by Rockenbach et al. (2011), the average antioxidant potency measured by the FRAP assay in the peel of red grape (Vitis vinifera) cvs. ‘Pinot Noir’, ‘Sangiovese’, and ‘Negro Amaro’ was by 58%, 49%, and 30% lower than in the peel of the Saskatoon berry genotypes investigated in our study. However, as reported by Pantelić et al. (2016) the antioxidant activity of grape flesh in cvs. ‘Pinot Noir’ and ‘Sangiovese’ was by 29% and 21% higher than that in the fruit flesh of the tested Saskatoon berry genotypes. It was proved that the total polyphenolics content played an important role in the antioxidant activity (r2 = 0.697 for FRAP and r2 = 0.815 for DPPH), in addition to individual groups of compounds.

3.4. Tetraterpenoids The quantitative analysis of tetraterpenoids in the fruit of the Saskatoon berry genotypes and their parts was determined using the LC/MS technique (Table 3), and demonstrated their contents to differ significantly depending on the analyzed part of fruit and also on genotype. The content of carotenoids in the fruit of the evaluated genotypes ranged from 262 mg/kg d.m. in cv. ‘Pembina’ to 517 mg/kg d.m. in 'clone type S'. Similar results were obtained for Saskatoon berry genotypes cultivated in Poland (Lachowicz et al., 2019; Lachowicz, Oszmiański, Seliga, & Pluta, 2017). Among the examined fruit parts of these genotypes, the peel was the richest in carotenoids, the average content of which reached 348 mg/kg d.m. and was 1.4 and 1.7 times 8

Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds

Thiessen

9

AC 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.0 0.1 0.1 0.1 0.1 0.1 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.1 0.0 0.1 0.0 0.1 0.0 0.0 0.0

TBC

9CBC 1.0 0.6 1.0 0.6 0.9 0.6 0.9 0.5 0.8 0.5 0.8 0.5 0.6 0.3 0.5 0.3 0.6 0.3 0.5 0.3 1.1 0.6 1.0 0.6 1.0 0.6 0.9 0.5

13CBC

TC

4.2 ± 5.2 ± 3.9 ± 1.5 ± 3.8 ± 4.7 ± 3.5 ± 1.4 ± 3.0 ± 3.7 ± 2.8 ± 1.1 ± 2.0 ± 2.5 ± 1.8 ± 0.7 ± 3.4 ± 4.2 ± 3.2 ± 1.3 ± 2.6 ± 3.2 ± 2.4 ± 1.0 ± 2.3 ± 2.9 ± 2.2 ± 0.9 ± 3.1b 3.8a 2.8c 1.1d

0.1a 0.1 0.1 0.0 0.1 0.1 0.1 0.0 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.1 0.1 0.0 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 152.0 ± 2.4 99.2 ± 1.6 107.6 ± 1.7 67.4 ± 1.1 138.6 ± 2.2 90.5 ± 1.4 98.1 ± 1.6 61.5 ± 1.0 116.8 ± 1.9 76.2 ± 1.2 82.7 ± 1.3 51.8 ± 0.8 77.5 ± 1.2 50.6 ± 0.8 54.8 ± 0.9 34.4 ± 0.5 82.5 ± 1.3 53.9 ± 0.9 58.4 ± 0.9 36.6 ± 0.6 148.2 ± 2.4 96.7 ± 1.5 104.9 ± 1.7 65.7 ± 1.1 132.2 ± 2.1 86.3 ± 1.4 93.6 ± 1.5 58.6 ± 0.9 121.1a 79.1c 85.7b 53.7d

4.3 ± 0.1 2.8 ± 0.0 3.0 ± 0.0 1.9 ± 0.0 3.9 ± 0.1 2.5 ± 0.0 2.8 ± 0.0 1.7 ± 0.0 4.2 ± 0.1 2.7 ± 0.0 3.0 ± 0.0 1.9 ± 0.0 2.8 ± 0.0 1.8 ± 0.0 2.0 ± 0.0 1.2 ± 0.0 7.4 ± 0.1 4.8 ± 0.1 5.2 ± 0.1 3.3 ± 0.1 11.6 ± 0.2 7.5 ± 0.1 8.2 ± 0.1 5.1 ± 0.1 10.3 ± 0.2 6.7 ± 0.1 7.3 ± 0.1 4.6 ± 0.1 6.3a 4.1c 4.5b 2.8d

5.8 ± 3.5 ± 5.4 ± 3.1 ± 5.3 ± 3.2 ± 5.0 ± 2.9 ± 6.8 ± 4.1 ± 6.3 ± 3.7 ± 4.5 ± 2.7 ± 4.2 ± 2.4 ± 1.6 ± 0.9 ± 1.5 ± 0.8 ± 3.7 ± 2.2 ± 3.4 ± 2.0 ± 3.3 ± 2.0 ± 3.0 ± 1.8 ± 4.4a 2.6c 4.1b 2.4d

241.5 ± 3.9 144.4 ± 2.3 225.3 ± 3.6 130.3 ± 2.1 220.2 ± 3.5 131.7 ± 2.1 205.4 ± 3.3 118.8 ± 1.9 205.6 ± 3.3 123.0 ± 2.0 191.8 ± 3.1 110.9 ± 1.8 136.3 ± 2.2 81.6 ± 1.3 127.2 ± 2.0 73.5 ± 1.2 133.2 ± 2.1 79.7 ± 1.3 124.3 ± 2.0 71.9 ± 1.2 265.8 ± 4.3 159.0 ± 2.5 248.0 ± 4.0 143.4 ± 2.3 237.1 ± 3.8 141.8 ± 2.3 221.2 ± 3.5 127.9 ± 2.0 205.7a 123.0c 191.9b 110.9d

63.9 ± 38.2 ± 59.6 ± 34.4 ± 58.2 ± 34.8 ± 54.3 ± 31.4 ± 52.7 ± 31.5 ± 49.1 ± 28.4 ± 34.9 ± 20.9 ± 32.6 ± 18.8 ± 34.4 ± 20.6 ± 32.1 ± 18.6 ± 67.1 ± 40.2 ± 62.7 ± 36.2 ± 59.9 ± 35.8 ± 55.9 ± 32.3 ± 53.0a 31.7c 49.5b 28.6d

2.1 ± 0.0 1.3 ± 0.0 2.0 ± 0.0 1.1 ± 0.0 1.9 ± 0.0 1.2 ± 0.0 1.8 ± 0.0 1.0 ± 0.0 6.8 ± 0.1 4.1 ± 0.1 6.4 ± 0.1 3.7 ± 0.1 4.5 ± 0.1 2.7 ± 0.0 4.2 ± 0.1 2.4 ± 0.0 18.7 ± 0.3 11.2 ± 0.2 17.5 ± 0.3 10.1 ± 0.2 20.1 ± 0.3 12.0 ± 0.2 18.7 ± 0.3 10.8 ± 0.2 17.9 ± 0.3 10.7 ± 0.2 16.7 ± 0.3 9.7 ± 0.2 10.3a 6.2c 9.6b 5.6d

473.7 ± 294.5 ± 406.8 ± 239.8 ± 431.9 ± 268.6 ± 370.9 ± 218.7 ± 395.9 ± 245.3 ± 342.1 ± 201.4 ± 262.5 ± 162.7 ± 226.9 ± 133.6 ± 281.3 ± 175.4 ± 242.2 ± 142.5 ± 519.0 ± 320.8 ± 448.3 ± 264.2 ± 463.0 ± 286.2 ± 399.9 ± 235.7 ± 403.9a 250.5c 348.2b 205.1d

7.6 4.7 6.5 3.8 6.9 4.3 5.9 3.5 6.3 3.9 5.5 3.2 4.2 2.6 3.6 2.1 4.5 2.8 3.9 2.3 8.3 5.1 7.2 4.2 7.4 4.6 6.4 3.8

0.7 ± 0.4 ± 0.3 ± 0.3 ± 0.7 ± 0.3 ± 0.2 ± 0.2 ± 0.2 ± 0.1 ± 0.1 ± 0.1 ± 0.1 ± 0.1 ± 0.1 ± 0.1 ± 2.6 ± 1.3 ± 0.9 ± 0.9 ± 1.2 ± 0.6 ± 0.4 ± 0.4 ± 1.1 ± 0.5 ± 0.4 ± 0.4 ± 1.0a 0.5b 0.3c 0.3c

0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

18.2 ± 0.3 8.9 ± 0.1 6.3 ± 0.1 6.4 ± 0.1 16.6 ± 0.3 8.1 ± 0.1 5.7 ± 0.1 5.9 ± 0.1 14.3 ± 0.2 7.0 ± 0.1 4.9 ± 0.1 5.1 ± 0.1 9.5 ± 0.2 4.6 ± 0.1 3.3 ± 0.1 3.4 ± 0.1 29.6 ± 0.5 14.5 ± 0.2 10.2 ± 0.2 10.4 ± 0.2 31.1 ± 0.5 15.2 ± 0.2 10.7 ± 0.2 11.0 ± 0.2 27.7 ± 0.4 13.6 ± 0.2 9.6 ± 0.2 9.8 ± 0.2 21.0a 10.3b 7.3d 7.4c

Cdb

Ppa

13CL

Ze

TL

Chlorophylls (mg/kg dm)

Carotenoids (mg/kg dm)

Tetraterpenoids

1.0 ± 0.5 ± 0.3 ± 0.4 ± 0.9 ± 0.4 ± 0.3 ± 0.3 ± 5.3 ± 2.6 ± 1.8 ± 1.9 ± 3.5 ± 1.7 ± 1.2 ± 1.3 ± 2.4 ± 1.2 ± 0.8 ± 0.8 ± 9.7 ± 4.7 ± 3.3 ± 3.4 ± 8.6 ± 4.2 ± 3.0 ± 3.1 ± 4.5a 2.2b 1.6c 1.6c

Cb 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.0 0.0 0.0 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.2 0.1 0.1 0.1 0.1 0.1 0.0 0.0

44.6 ± 35.1 ± 93.2 ± 25.0 ± 40.7 ± 32.0 ± 84.9 ± 22.8 ± 37.5 ± 29.6 ± 78.4 ± 21.1 ± 24.9 ± 19.6 ± 52.0 ± 14.0 ± 23.1 ± 18.2 ± 48.2 ± 13.0 ± 43.1 ± 33.9 ± 90.0 ± 24.2 ± 38.4 ± 30.3 ± 80.3 ± 21.6 ± 36.0 28.4c 75.3a 20.2d

Pa 0.7 0.6 1.5 0.4 0.7 0.5 1.4 0.4 0.6 0.5 1.3 0.3 0.4 0.3 0.8 0.2 0.4 0.3 0.8 0.2 0.7 0.5 1.4 0.4 0.6 0.5 1.3 0.3

6.8 ± 0.1 5.4 ± 0.1 14.3 ± 0.2 3.8 ± 0.1 6.2 ± 0.1 4.9 ± 0.1 13.0 ± 0.2 3.5 ± 0.1 9.5 ± 0.2 7.5 ± 0.1 19.8 ± 0.3 5.3 ± 0.1 6.3 ± 0.1 4.9 ± 0.1 13.1 ± 0.2 3.5 ± 0.1 22.7 ± 0.4 17.9 ± 0.3 47.5 ± 0.8 12.8 ± 0.2 18.8 ± 0.3 14.8 ± 0.2 39.4 ± 0.6 10.6 ± 0.2 16.8 ± 0.3 13.2 ± 0.2 35.1 ± 0.6 9.4 ± 0.2 12.4b 9.8d 26.0a 7.0c

Pb

71.4 ± 1.1 50.2 ± 0.8 114.3 ± 1.8 35.9 ± 0.6 65.1 ± 1.0 45.8 ± 0.7 104.2 ± 1.7 32.7 ± 0.5 66.9 ± 1.1 46.7 ± 0.7 105.0 ± 1.7 33.4 ± 0.5 44.3 ± 0.7 31.0 ± 0.5 69.7 ± 1.1 22.2 ± 0.4 80.4 ± 1.3 53.0 ± 0.8 107.6 ± 1.7 37.9 ± 0.6 103.9 ± 1.7 69.3 ± 1.1 143.9 ± 2.3 49.6 ± 0.8 92.7 ± 1.5 61.9 ± 1.0 128.4 ± 2.1 44.3 ± 0.7 74.9b 51.1c 110.4a 36.6d

TCh

a Values are means ± standard deviation, a–d Means - SD followed by different letters within the same line represent significant differences (p < 0.05). n = 3; Explanations: Ze zeoxanthin; TL all-trans-lutein; 13CL 13cis-lutein; AC α-carotene; all-trans-β-carotene; 9CBC 9-cis-β-carotene; 13CBC 13-cis-β-carotene; TC total carotenoids; Ppa Pheophorbide; Cpb chlorophyllide b; Cb chlorophyll b; Pa Pheophytin a; Pb pheophytin b.

Mean

Clone type N

Clone type S

Clone no 5/6

Pembina

Martin

Smoky

Their components

Genotypes

Table 3 The content of tetraterpenoids in seven Saskatoon berry genotypes and their parts.

S. Lachowicz, et al.

Food Chemistry 305 (2020) 125430

Food Chemistry 305 (2020) 125430

S. Lachowicz, et al.

Table 4 The content of triterpenoids and antioxidant activity in seven Saskatoon berry genotypes and their parts. Genotypes

Their components

Triterpenoids [mg/kg d.m.] Betulinic acid

Thiessen

Smoky

Martin

Pembina

Clone no 5/6

Clone type S

Clone type N

Mean

a

Fruit Flesh Peel Seeds Fuit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds Fruit Flesh Peel Seeds

a

14.9 ± 0.3 9.7 ± 0.2 14.3 ± 0.3 12.8 ± 0.3 10.8 ± 0.2 4.5 ± 0.1 13.7 ± 0.3 8.3 ± 0.2 13.5 ± 0.3 9.0 ± 0.2 22.7 ± 0.5 10.5 ± 0.2 7.2 ± 0.1 3.6 ± 0.1 3.9 ± 0.1 1.4 ± 0.0 13.5 ± 0.3 3.7 ± 0.1 27.5 ± 0.6 17.5 ± 0.3 12.7 ± 0.3 7.3 ± 0.1 6.4 ± 0.1 6.7 ± 0.1 11.9 ± 0.2 2.5 ± 0.1 8.5 ± 0.2 5.9 ± 0.1 12.1b 5.8d 13.9a 9.0c

Antioxidant capacity [mmol TE/100 g d.m.]

Ursolic acid

Oleanolic acid

SUM

FRAP

DPPH

36.6 ± 0.7 10.8 ± 0.2 44.9 ± 0.9 11.6 ± 0.2 16.0 ± 0.3 24.2 ± 0.5 42.9 ± 0.9 25.8 ± 0.5 19.6 ± 0.4 9.4 ± 0.2 66.7 ± 1.3 25.6 ± 0.5 19.3 ± 0.4 4.1 ± 0.1 22.0 ± 0.4 3.0 ± 0.1 30.5 ± 0.6 16.1 ± 0.3 31.7 ± 0.6 72.5 ± 1.4 5.6 ± 0.1 20.5 ± 0.4 28.4 ± 0.6 16.9 ± 0.3 29.5 ± 0.6 3.6 ± 0.1 23.3 ± 0.5 7.7 ± 0.2 22.5b 12.7d 37.1a 23.3c

20.2 ± 0.4 23.8 ± 0.5 7.0 ± 0.1 11.1 ± 0.2 21.7 ± 0.4 2.6 ± 0.1 51.7 ± 1.0 19.8 ± 0.4 26.2 ± 0.5 28.9 ± 0.6 26.8 ± 0.5 7.5 ± 0.1 5.2 ± 0.1 1.7 ± 0.0 4.2 ± 0.1 2.0 ± 0.0 6.9 ± 0.1 15.5 ± 0.3 154.3 ± 3.1 65.9 ± 1.3 15.5 ± 0.3 5.9 ± 0.1 40.4 ± 0.8 10.1 ± 0.2 10.4 ± 0.2 6.5 ± 0.1 4.6 ± 0.1 3.4 ± 0.1 15.1c 12.1d 41.3a 17.1b

71.7 ± 1.4 44.3 ± 0.9 66.2 ± 1.3 35.5 ± 0.7 48.5 ± 1.0 31.3 ± 0.6 108.3 ± 2.2 31.3 ± 0.6 59.3 ± 1.2 47.4 ± 0.9 116.1 ± 2.3 43.6 ± 0.9 31.7 ± 0.6 79.9 ± 1.6 30.1 ± 0.6 6.3 ± 0.1.0 50.9 ± 1.0 35.3 ± 0.7 213.5 ± 4.3 155.8 ± 3.1 33.8 ± 0.7 33.7 ± 0.7 75.2 ± 1.5 91.9 ± 1.8 51.8 ± 1.0 12.7 ± 0.3 36.4 ± 0.7 17.0 ± 0.3 49.7c 40.6d 92.3a 54.5b

17.0 ± 0.0 6.8 ± 0.0 41.5 ± 0.1 2.1 ± 0.0 18.9 ± 0.0 6.9 ± 0.0 30.2 ± 0.1 2.0 ± 0.0 15.2 ± 0.0 8.3 ± 0.0 25.1 ± 0.1 1.1 ± 0.0 11.4 ± 0.0 2.3 ± 0.0 14.5 ± 0.0 0.8 ± 0.0 9.5 ± 0.0 2.8 ± 0.0 20.4 ± 0.0 0.2 ± 0.0 17.9 ± 0.0 4.5 ± 0.0 36.3 ± 0.1 3.6 ± 0.0 15.9 ± 0.0 6.2 ± 0.0 36.6 ± 0.1 2.7 ± 0.0 15.1b 5.4c 29.2a 1.8d

27.0 ± 0.2 10.7 ± 0.1 65.7 ± 0.4 3.4 ± 0.0 29.9 ± 0.2 10.9 ± 0.1 47.8 ± 0.3 3.1 ± 0.0 24.0 ± 0.1 13.1 ± 0.1 39.7 ± 0.2 1.8 ± 0.0 18.1 ± 0.1 3.6 ± 0.0 23.0 ± 0.1 1.2 ± 0.0 15.0 ± 0.1 4.4 ± 0.0 32.3 ± 0.2 0.4 ± 0.0 28.3 ± 0.2 7.2 ± 0.0 57.4 ± 0.3 5.7 ± 0.0 25.1 ± 0.2 9.9 ± 0.1 58.0 ± 0.3 4.3 ± 0.0 23.9b 8.5c 46.3a 2.8d

Values are means ± standard deviation, a–d Means - SD followed by different letters within the same line represent significant differences (p < 0.05). n = 3.

The antioxidant activity of the fruit of the analyzed genotypes and their parts was more strongly positively correlated with the content of polymeric procyanidins, anthocyanins, and phenolic acids (r2 = 0.805, r2 = 0.791 and r2 = 0.666 for FRAP and r2 = 0.760, r2 = 0.702 and r2 = 0.804 for DPPH) than with contents of flavanols and flavonols (r2 = 0.479 and r2 = 0.173 for FRAP and r2 = 0.531 and r2 = 0.320 for DPPH). In addition to polyphenolic compounds, positive correlations were reported between antioxidative potency and carotenoids (r2 = 0.599 for FRAP, and r2 = 0.671 for DPPH). In turn, Loza-Mejía and Salazar (2015) noted that the antioxidative properties of the analyzed berries were positively correlated with the total triterpenoids content (r2 = 0.527 for FRAP and r2 = 0.621 for DPPH). Besides, the antioxidant properties of fruits of the tested genotypes and particular fruit parts depended on the composition of health-promoting components including polyphenols and terpenoids (Bakowska-Barczak & Kolodziejczyk, 2008).

flavonols; fruit seeds – with the highest content of polymeric procyanidins; and fruit peel – with the highest contents of pectins, phenolic acids, anthocyanins, terpenoids, and triterpenoids; and a strong positive correlation between results of the FRAP and DPPH assays. The PCA illustrated also that the genotype affected contents of polyphenolic compounds and their groups to a greater extent than contents of the other analyzed compounds. It was reflected in the content of these compounds in fruit parts. The obtained results showed how individual polyphenolic and terpenoid compounds were distributed in particular parts of the fruit and how many valuable compounds remained in the post-production pomace. Moreover, the pomace contained mainly seeds and peel, and also some minor amounts of fruit flesh. 4. Conclusions The present study provides valid information on the health properties of peel, flesh, and seeds isolated from fruit of seven different Saskatoon berry genotypes cultivated in Poland, related to their triterpenoid, tetraterpenoid, a polyphenol profiles, and to their antioxidative potency. Furthermore, results of the presented analyses indicate statistically significant differences between particular fruit parts and Saskatoon berry genotypes. The fruit peel of the following genotypes: ‘Thiessen, 'type S', and 'type N', contained mainly anthocyanins, polymerized compounds, hydroxycinnamic acids, triterpenoids, and carotenoids, as well as exhibited antioxidant activity. In turn,

3.6. Principal component analysis (PCA) Results concerning contents terpenoids, polyphenols, and antioxidants in the fruit of the tested selected genotypes and their parts (peel, flesh, and seeds) were analyzed using the PCA. Two major principal components, i.e. PC1 (45.35%) and PC2 (28.95%), were illustrated in Fig. 1. The obtained data show the presence of 3 groups: fruit flesh – with the highest contents of soluble solid, ash, and

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S. Lachowicz, et al.

PCA 1 v 2 (74.30%) 1,0

Ash Soluble solids

Flavonols

0,8 0,6

Pectins Clone no 5/6(F) Martin(F) Pembina(F) Smoky(F) Clone Type S(F) Thessien(F) Flavanols Catotenoids Clone Type S(P) Clone Type N(F) Chlorophylls Phenolic acids Pembina(P) Smoky(P) DPPH Clone no 5/6(P) FRAP Thessien(P) Anthocyanins Martin(P) Clone Type S(S) Oleanolic acid Phenolic compounds Thessien(S) Martin(S) Clone Type N(P) Smoky(S) Ursolic acid Pembina(S) Clone no 5/6(S) Triterpenoids Clone Type N(S) Betulinic acid

PC2 (28.95%)

0,4 0,2 0,0 -0,2 -0,4 -0,6

Polimeric procyanidins

-0,8 -1,0 -0,8

-0,6

-0,4

-0,2

0,0

0,2

0,4

0,6

0,8

1,0

1,2

PC1 (45.35%) Fig. 1. Principal components analysis showing the relationship among chemical composition and antioxidants in peel (P), flesh (F) and seeds (S) of seven Saskatoon berry genotypes.

anthocyanins and polymeric procyanidins were determined in the peel of fruit of cvs. ‘Smoky’ and ‘Martin’, whereas flavonols predominated in the flesh, and polymerized compounds in the seeds. In the case of the remaining genotypes, significant amounts of flavonols were found in fruit flesh, whereas – these of polymeric procyanidins in the seeds. Particular parts, i.e. peel, flesh, and seeds, of the fruit of Saskatoon berry are valuable waste products. They contain many health-promoting compounds and can be good raw materials to manufacture functional foods or dietary supplements.

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