J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Original Research

Volume 13, March 2021, pages 52-61

Polyphenols, vitamin C, and sugar in commercial apple beverages and their effects on antioxidant and anti-inflammatory activities in vitro

Figure 1. (a) 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, (b) inhibitions of formation of F₂-isoprostanes, (c) lipid hydroperoxides, (d) leukotriene B₄, and (e) myeloperoxidase activity by freshly isolated human neutrophils in vitro of apple beverages – juices and drinks with low (lower than 10 mg GAE/100g; apple beverages n = 8, juices n = 6, drinks n = 2) and high (equal or greater than 10 mg GAE/100g; apple beverages n = 9, juices n = 6, drinks n = 3) total polyphenol contents. *p < 0.05 using two-sample independent t-test.

Figure 2. (a) 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, (b) inhibitions of formation of F₂-isoprostanes, (c) lipid hydroperoxides, (d) leukotriene B₄, and (e) myeloperoxidase activity by freshly isolated human neutrophils in vitro of apple beverages – juices and drinks with low (lower than 1.2 mg/100g; apple beverages n = 9, juices n = 6, drinks n = 3) and high (equal or greater than 1.2 mg/100g; apple beverages n = 8, juices n = 6, drinks n = 2) total catechin contents. *p < 0.05 using two-sample independent t-test.

Figure 3. (a) 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, (b) inhibitions of formation of F₂-isoprostanes, (c) lipid hydroperoxides, (d) leukotriene B₄, and (e) myeloperoxidase activity by freshly isolated human neutrophils in vitro of apple beverages – juices and drinks with low (lower than 0.5 mg/100g; apple beverages n = 10, juices n = 7, drinks n = 3) and high (equal or greater than 0.5 mg/100g; apple beverages n = 7, juices n = 5, drinks n = 2) total quercetin contents. *p < 0.05 using two-sample independent t-test.

Figure 4. (a) 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, (b) inhibitions of formation of F₂-isoprostanes, (c) lipid hydroperoxides, (d) leukotriene B₄, and (e) myeloperoxidase activity by freshly isolated human neutrophils in vitro of apple beverages – juices and drinks with low (lower than 20 mg/100g; apple beverages n = 7, juices n = 7, drinks n = 0) and high (equal or greater than 20 mg/100g; apple beverages n = 10, juices n = 5, drinks n = 5) vitamin C. *p < 0.05 using two-sample independent t-test.

**Table 1.** Nutritional profile, total and specific polyphenol contents of apple beverages (N¹)
Nutrients	All	Juices	Drinks
¹N represents the number of fruit beverage products out of the 17 apple beverages with declared respective nutrient contents on their product labels. p < 0.05 vs. juices using two-sample independent t-tests. *p < 0.05 vs. healthy 5.0 g/100 mL reference sugar value using one-sample t-tests.
Energy (N) (kJ/100g)	45.07 ± 3.74 (17)	46.08 ± 2.28 (12)	42.65 ± 5.58* (5)
Protein (N) (g/100g)	0.10 ± 0.12 (17)	0.11 ± 0.13 (12)	0.09 ± 0.09 (5)
Fat (N) (g/100g)	0.03 ± 0.07 (17)	0.03 ± 0.08 (12)	0.02 ± 0.05 (5)
Carbohydrate (N) (g/100g)	11.08 ± 1.03 (17)	11.34 ± 0.68 (12)	10.43 ± 1.50 (5)
Sugar (N) (g/100g)	10.35 ± 1.18** (17)	10.64 ± 0.88** (12)	9.65 ± 1.61* ** (5)
Dietary fibre (N) (g/100g)	0.01 ± 0.04 (9)	0.00 ± 0.00 (6)	0.04 ± 0.07 (3)
Sodium (N) (mg/100g)	6.78 ± 8.55 (16)	4.73 ± 5.91 (12)	12.94 ± 13.05 (4)
Vitamin C (N) (mg/100g)	33.35 ± 21.87 (17)	19.50 ± 9.88 (12)	44.42 ± 23.19 (5)*
Total polyphenols (N) (mg gallic acid equivalent/100g)	9.9 ± 1.2 (17)	10.7 ± 1.2 (12)	9.4 ± 0.8 (5)*
Total catechin (mg/100g)	1.60 ± 0.10 (17)	1.65 ± 0.07 (12)	1.50 ± 0.08 (5)*
Total quercetin (mg/100g)	0.46 ± 0.09 (17)	0.50 ± 0.06 (12)	0.36 ± 0.07 (5)*

Table 1. Nutritional profile, total and specific polyphenol contents of apple beverages (N¹)

Nutrients

All

Juices

Drinks

¹N represents the number of fruit beverage products out of the 17 apple beverages with declared respective nutrient contents on their product labels. *p < 0.05 vs. juices using two-sample independent t-tests. **p < 0.05 vs. healthy 5.0 g/100 mL reference sugar value using one-sample t-tests.

Energy (N) (kJ/100g)

45.07 ± 3.74 (17)

46.08 ± 2.28 (12)

42.65 ± 5.58* (5)

Protein (N) (g/100g)

0.10 ± 0.12 (17)

0.11 ± 0.13 (12)

0.09 ± 0.09 (5)

Fat (N) (g/100g)

0.03 ± 0.07 (17)

0.03 ± 0.08 (12)

0.02 ± 0.05 (5)

Carbohydrate (N) (g/100g)

11.08 ± 1.03 (17)

11.34 ± 0.68 (12)

10.43 ± 1.50 (5)

Sugar (N) (g/100g)

10.35 ± 1.18** (17)

10.64 ± 0.88** (12)

9.65 ± 1.61* ** (5)

Dietary fibre (N) (g/100g)

0.01 ± 0.04 (9)

0.00 ± 0.00 (6)

0.04 ± 0.07 (3)

Sodium (N) (mg/100g)

6.78 ± 8.55 (16)

4.73 ± 5.91 (12)

12.94 ± 13.05 (4)

Vitamin C (N) (mg/100g)

33.35 ± 21.87 (17)

19.50 ± 9.88 (12)

44.42 ± 23.19 (5)*

Total polyphenols (N) (mg gallic acid equivalent/100g)

9.9 ± 1.2 (17)

10.7 ± 1.2 (12)

9.4 ± 0.8 (5)*

Total catechin (mg/100g)

1.60 ± 0.10 (17)

1.65 ± 0.07 (12)

1.50 ± 0.08 (5)*

Total quercetin (mg/100g)

0.46 ± 0.09 (17)

0.50 ± 0.06 (12)

0.36 ± 0.07 (5)*

**Table 2.** Antioxidant and anti-inflammatory capacity of apple beverages (N¹)
Nutrients	All	Juices	Drinks
¹N represents the number of fruit beverage products out of the 17 apple beverages with declared respective nutrient contents on their product labels. *p < 0.05 vs. juices using two-sample independent t-tests.
1,1-diphenyl-2-picrylhydrazyl radical scavenging antioxidant capacity (N) (mg vitamin C equivalents/100g)	0.64 ± 0.14 (17)	0.56 ± 0.05 (12)	0.85 ± 0.06 (5)*
Inhibition of cellular F₂-isoprostane production (N) (% relative to the positive control)	34.1 ± 3.7 (17)	33.1 ± 3.8 (12)	36.6 ± 1.6 (5)*
Inhibition of cellular lipid hydroperoxide production (N) (% relative to the positive control)	18.0 ± 1.6 (17)	17.6 ± 1.6 (12)	19.1 ± 0.9 (5)*
Inhibition of cellular leukotriene B₄ production (N) (% relative to the positive control)	24.0 ± 1.14 (17)	24.3 ± 0.1 (12)	23.1 ± 1.1 (5)*
Inhibition of cellular myeloperoxidase activity (N) (% relative to the positive control)	25.2 ± 1.4 (17)	25.1 ± 1.5 (12)	23.1 ± 2.6 (5)*

Table 2. Antioxidant and anti-inflammatory capacity of apple beverages (N¹)

Nutrients

All

Juices

Drinks

1,1-diphenyl-2-picrylhydrazyl radical scavenging antioxidant capacity (N) (mg vitamin C equivalents/100g)

0.64 ± 0.14 (17)

0.56 ± 0.05 (12)

0.85 ± 0.06 (5)*

Inhibition of cellular F₂-isoprostane production (N) (% relative to the positive control)

34.1 ± 3.7 (17)

33.1 ± 3.8 (12)

36.6 ± 1.6 (5)*

Inhibition of cellular lipid hydroperoxide production (N) (% relative to the positive control)

18.0 ± 1.6 (17)

17.6 ± 1.6 (12)

19.1 ± 0.9 (5)*

Inhibition of cellular leukotriene B₄ production (N) (% relative to the positive control)

24.0 ± 1.14 (17)

24.3 ± 0.1 (12)

23.1 ± 1.1 (5)*

Inhibition of cellular myeloperoxidase activity (N) (% relative to the positive control)

25.2 ± 1.4 (17)

25.1 ± 1.5 (12)

23.1 ± 2.6 (5)*

**Table 3.** Antioxidant and anti-inflammatory capacity of simulated apple beverages
	AL¹	ALS²	AH³	AHS⁴	AB⁵	ABH⁶
¹AL represented a simulated apple beverage containing vitamin C (20 mg/100g), apple polyphenols (10 mg gallic acid equivalent/100g), catechin (1.2 mg/100g), and quercetin (0.5 mg/100g). ²ALS represented a simulated apple beverage containing vitamin C (20 mg/100g), apple polyphenols (10 mg gallic acid equivalent/100g), catechin (1.2 mg/100g), quercetin (0.5 mg/100g), and fructose (10 g/100g). ³AH represented a simulated apple beverage containing vitamin C (80 mg/100g), apple polyphenols (40 mg gallic acid equivalent/100g), catechin (5 mg/100g), and quercetin (2 mg/100g). ⁴AHS represented a simulated apple beverage containing vitamin C (80 mg/100g), apple polyphenols (40 mg gallic acid equivalent/100g), catechin (5 mg/100g), quercetin (2 mg/100g), and fructose (10 g/100g). ⁵AB was one of the studied commercial apple beverage measured to contain vitamin C (20.5 mg/100g), apple polyphenols (9.5 mg GAE/100g), catechin (1.35 mg/100g), quercetin (0.42 mg/100g), and fructose (10 g/100g). ⁶ABH represented AB and added vitamin C (59.5 mg/100g), apple polyphenols (30.5 mg GAE/100g), catechin (3.65 mg/100g), and quercetin (1.58 mg/100g). ^a,bdifferent alphabets represent significant difference using ANOVA with Bonferroni adjustment.
1,1-diphenyl-2-picrylhydrazyl radical scavenging antioxidant capacity (N) (mg vitamin C equivalents/100g)	1.02 ± 0.08^a	1.03 ± 0.09^a	3.65 ± 0.08^b	3.79 ± 0.16^b	0.52 ± 0.03^a	3.35 ± 0.19^b
Inhibition of cellular F₂-isoprostane production (N) (% relative to the positive control)	33.7 ± 0.8^a	32.2 ± 1.6^a	53.6 ± 2.7^b	57.3 ± 3.1^b	35.2 ± 2.1^a	59.1 ± 1.1^b
Inhibition of cellular lipid hydroperoxide production (N) (% relative to the positive control)	17.0 ± 1.6^a	18.0 ± 1.5^a	32.4 ± 3.1^b	29.8 ± 1.3^b	18.2 ± 0.3^a	30.6 ± 1.7^b
Inhibition of cellular leukotriene B₄ production (N) (% relative to the positive control)	25.8 ± 3.2^a	26.1 ± 3.0^a	48.7 ± 4.0^b	46.4 ± 1.6^b	25.8 ± 1.3^a	48.9 ± 1.4^b
Inhibition of cellular myeloperoxidase activity (N) (% relative to the positive control)	25.2 ± 1.5^a	24.9 ± 2.2^a	40.7 ± 0.3^b	42.1 ± 1.9^b	24.9 ± 3.4^a	45.1 ± 4.6^b

Table 3. Antioxidant and anti-inflammatory capacity of simulated apple beverages

AL¹

ALS²

AH³

AHS⁴

AB⁵

ABH⁶

¹AL represented a simulated apple beverage containing vitamin C (20 mg/100g), apple polyphenols (10 mg gallic acid equivalent/100g), catechin (1.2 mg/100g), and quercetin (0.5 mg/100g). ²ALS represented a simulated apple beverage containing vitamin C (20 mg/100g), apple polyphenols (10 mg gallic acid equivalent/100g), catechin (1.2 mg/100g), quercetin (0.5 mg/100g), and fructose (10 g/100g). ³AH represented a simulated apple beverage containing vitamin C (80 mg/100g), apple polyphenols (40 mg gallic acid equivalent/100g), catechin (5 mg/100g), and quercetin (2 mg/100g). ⁴AHS represented a simulated apple beverage containing vitamin C (80 mg/100g), apple polyphenols (40 mg gallic acid equivalent/100g), catechin (5 mg/100g), quercetin (2 mg/100g), and fructose (10 g/100g). ⁵AB was one of the studied commercial apple beverage measured to contain vitamin C (20.5 mg/100g), apple polyphenols (9.5 mg GAE/100g), catechin (1.35 mg/100g), quercetin (0.42 mg/100g), and fructose (10 g/100g). ⁶ABH represented AB and added vitamin C (59.5 mg/100g), apple polyphenols (30.5 mg GAE/100g), catechin (3.65 mg/100g), and quercetin (1.58 mg/100g). ^a,bdifferent alphabets represent significant difference using ANOVA with Bonferroni adjustment.

1,1-diphenyl-2-picrylhydrazyl radical scavenging antioxidant capacity (N) (mg vitamin C equivalents/100g)

1.02 ± 0.08^a

1.03 ± 0.09^a

3.65 ± 0.08^b

3.79 ± 0.16^b

0.52 ± 0.03^a

3.35 ± 0.19^b

Inhibition of cellular F₂-isoprostane production (N) (% relative to the positive control)

33.7 ± 0.8^a

32.2 ± 1.6^a

53.6 ± 2.7^b

57.3 ± 3.1^b

35.2 ± 2.1^a

59.1 ± 1.1^b

Inhibition of cellular lipid hydroperoxide production (N) (% relative to the positive control)

17.0 ± 1.6^a

18.0 ± 1.5^a

32.4 ± 3.1^b

29.8 ± 1.3^b

18.2 ± 0.3^a

30.6 ± 1.7^b

Inhibition of cellular leukotriene B₄ production (N) (% relative to the positive control)

25.8 ± 3.2^a

26.1 ± 3.0^a

48.7 ± 4.0^b

46.4 ± 1.6^b

25.8 ± 1.3^a

48.9 ± 1.4^b

Inhibition of cellular myeloperoxidase activity (N) (% relative to the positive control)

25.2 ± 1.5^a

24.9 ± 2.2^a

40.7 ± 0.3^b

42.1 ± 1.9^b

24.9 ± 3.4^a

45.1 ± 4.6^b