J Food Bioact

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

Journal website www.isnff-jfb.com

Original Research

Volume 14, June 2021, pages 114-125

Functional fermented carob milk: probiotic viability and polyphenolic profile

Figure 1. Changes in total phenolic content (TPC) of carob fermented milk (FCP12) during refrigerated storage (a) and in vitro digestion (b). Small letters indicate significant differences (p < 0.05) during storage period and different steps of digestion of the fermented milk. AF: After fermentation (16 h), BD: Before digestion.

Figure 2. Antioxidant activity (DPPH and ORAC) of carob fermented milk during the refrigerated storage (4 °C) (a, b) and during the in vitro digestion (c, d). Small letters indicate significant differences (p < 0.05) during storage period and different steps of digestion of the fermented milk. AF: after fermentation (16 h), BD: before digestion.

Figure 3. Survival of both probiotic strain (Lb. brevis B13 and B38) in carob fermented milk (FCP12) during the simulated gastrointestinal digestion. Small letters indicate a statistically significant difference between before the in vitro digestion counts and counts at each digestion step.

Figure 4. α-amylase (a) and α-glucosidase (b) inhibition (%) of carob fermented milk (FCP12) and its digested fractions. Small letters indicate significant differences before and during digestion steps for each tested concentration (0.3, 0.4, 0.5, 0.8 and 1 mg/mL).

**Table 1.** Experimental design for carob powder addition in fermented milks
Formulation	Prebiotic	Sample coding
SM: skim milk, F: fermented milk with only yogurt starter cultures, C: carob powder, I: inulin, FC: fermented milk with carob powder, FI: fermented milk with Inulin, P1: probiotic 1 (Lb. brevis B13), P2: probiotic 2 (Lb. brevis B38), P12: P1 + P2. First column represents the recipe of 4 different treatments for each row; control group (top row), Carob powder added milk (middle row) and inulin added milk (last row). The second column represents what type of prebiotic addition to the formulations; “0” means no addition, “C” means carob powder addition, and “I” means inulin addition. The third column represents the coding for 4 different treatments for each sample group in 3 rows.
SM + Lc. lactis C15	0	F
F + Lb. brevis B13	0	FP1
F + Lb. brevis B38	0	FP2
F + Lb. brevis B13 + Lb. brevis B38	0	FP12
SM + Lc. lactis C15	C	FC
F + Lb. brevis B13	C	FCP1
F + Lb. brevis B38	C	FCP2
F + Lb. brevis B13 + Lb. brevis B38	C	FCP12
SM + Lc. lactis C15	I	FI
F + Lb. brevis B13	I	FIP1
F + Lb. brevis B38	I	FIP2
F + Lb. brevis B13 + Lb. brevis B38	I	FIP12

Table 1. Experimental design for carob powder addition in fermented milks

Formulation

Prebiotic

Sample coding

SM: skim milk, F: fermented milk with only yogurt starter cultures, C: carob powder, I: inulin, FC: fermented milk with carob powder, FI: fermented milk with Inulin, P1: probiotic 1 (Lb. brevis B13), P2: probiotic 2 (Lb. brevis B38), P12: P1 + P2. First column represents the recipe of 4 different treatments for each row; control group (top row), Carob powder added milk (middle row) and inulin added milk (last row). The second column represents what type of prebiotic addition to the formulations; “0” means no addition, “C” means carob powder addition, and “I” means inulin addition. The third column represents the coding for 4 different treatments for each sample group in 3 rows.

SM + Lc. lactis C15

F + Lb. brevis B13

FP1

F + Lb. brevis B38

FP2

F + Lb. brevis B13 + Lb. brevis B38

FP12

SM + Lc. lactis C15

F + Lb. brevis B13

FCP1

F + Lb. brevis B38

FCP2

F + Lb. brevis B13 + Lb. brevis B38

FCP12

SM + Lc. lactis C15

F + Lb. brevis B13

FIP1

F + Lb. brevis B38

FIP2

F + Lb. brevis B13 + Lb. brevis B38

FIP12

**Table 2.** Probiotic *Lb. brevis* (P1 and P2) viable counts (log CFU/mL) in fermented milks during storage period (4 °C)*
Fermented milks	Day 0	Day 1	Day 7	Day 14	Day 21	Day 28
*Values expressed in log CFU/mL as mean ± standard deviation (n = 3).^A,B,C Different capital letters in a column indicates significant differences between fermented milk supplemented with different prebiotics (P < 0.05); ^a,b,c,d Different lowercase letters in a row for each fermented milk denote significant differences during storage period (P < 0.05). F: fermented milk without prebiotic addition including P1/P2, FC: fermented milk with carob powder including P1/P2, FI: fermented milk with inulin powder including P1/P2.
FP1	6.43 ± 0.11^aA	6.42 ± 0.09^aA	6.35 ± 0.08^abA	6.28 ± 0.09^bA	6.17 ± 0.04^cA	6.04 ± 0.25^dA
FP2	6.39 ± 0.09^aA	6.40 ± 0.10^aA	6.38 ± 0.12^aA	6.21 ± 0.10^bA	6.01 ± 0.04^cA	5.92 ± 0.05^cA
FP12	6.28 ± 0.12^aA	6.26 ± 0.08^aA	6.11 ± 0.25^bA	6.05 ± 0.04^bA	5.82 ± 0.13^cA	5.74 ± 0.12^cA
FCP1	8.68 ± 0.12^aB	8.69 ± 0.10^aB	8.80 ± 0.13^bB	8.79 ± 0.04^bB	8.69 ± 0.09^bB	8.28 ± 0.09^cB
FCP2	8.61 ± 0.18^aB	8.60 ± 0.09^aB	8.79 ± 0.08^bB	8.81 ± 0.11^bB	8.70 ± 0.09^bB	8.26 ± 0.14^cB
FCP12	8.66 ± 0.11^aB	8.65 ± 0.05^aB	8.98 ± 0.08^bB	8.93 ± 0.12^bB	8.89 ± 0.25^bB	8.58 ± 0.21^cB
FIP1	7.67 ± 0.05^aC	7.65 ± 0.05^aC	7.57 ± 0.09^aC	7.41 ± 0.05^abC	7.30 ± 0.25^bC	7.27 ± 0.07^bC
FIP2	7.69 ± 0.18^aC	7.70 ± 0.11^aC	7.64 ± 0.12^aC	7.47 ± 0.12^bC	7. 26 ± 0.07^cC	6.81 ± 0.14^dC
FP12	7.61 ± 0.08^aC	7.60 ± 0.11^aC	7.53 ± 0.21^aC	7.46 ± 0.13^aC	6.82 ± 0.15^bC	6.43 ± 0.05^cC

Table 2. Probiotic Lb. brevis (P1 and P2) viable counts (log CFU/mL) in fermented milks during storage period (4 °C)*

Fermented milks

Day 0

Day 1

Day 7

Day 14

Day 21

Day 28

*Values expressed in log CFU/mL as mean ± standard deviation (n = 3).^A,B,C Different capital letters in a column indicates significant differences between fermented milk supplemented with different prebiotics (P < 0.05); ^a,b,c,d Different lowercase letters in a row for each fermented milk denote significant differences during storage period (P < 0.05). F: fermented milk without prebiotic addition including P1/P2, FC: fermented milk with carob powder including P1/P2, FI: fermented milk with inulin powder including P1/P2.

FP1

6.43 ± 0.11^aA

6.42 ± 0.09^aA

6.35 ± 0.08^abA

6.28 ± 0.09^bA

6.17 ± 0.04^cA

6.04 ± 0.25^dA

FP2

6.39 ± 0.09^aA

6.40 ± 0.10^aA

6.38 ± 0.12^aA

6.21 ± 0.10^bA

6.01 ± 0.04^cA

5.92 ± 0.05^cA

FP12

6.28 ± 0.12^aA

6.26 ± 0.08^aA

6.11 ± 0.25^bA

6.05 ± 0.04^bA

5.82 ± 0.13^cA

5.74 ± 0.12^cA

FCP1

8.68 ± 0.12^aB

8.69 ± 0.10^aB

8.80 ± 0.13^bB

8.79 ± 0.04^bB

8.69 ± 0.09^bB

8.28 ± 0.09^cB

FCP2

8.61 ± 0.18^aB

8.60 ± 0.09^aB

8.79 ± 0.08^bB

8.81 ± 0.11^bB

8.70 ± 0.09^bB

8.26 ± 0.14^cB

FCP12

8.66 ± 0.11^aB

8.65 ± 0.05^aB

8.98 ± 0.08^bB

8.93 ± 0.12^bB

8.89 ± 0.25^bB

8.58 ± 0.21^cB

FIP1

7.67 ± 0.05^aC

7.65 ± 0.05^aC

7.57 ± 0.09^aC

7.41 ± 0.05^abC

7.30 ± 0.25^bC

7.27 ± 0.07^bC

FIP2

7.69 ± 0.18^aC

7.70 ± 0.11^aC

7.64 ± 0.12^aC

7.47 ± 0.12^bC

7. 26 ± 0.07^cC

6.81 ± 0.14^dC

FP12

7.61 ± 0.08^aC

7.60 ± 0.11^aC

7.53 ± 0.21^aC

7.46 ± 0.13^aC

6.82 ± 0.15^bC

6.43 ± 0.05^cC

**Table 3.** The changes of physico-chemical properties of different fermented milks during refrigerated storage*
Fermented milks	pH	Titrable acidity (%)	Viscosity (cP)
*Values were expressed as mean ± standard deviation (n = 3). ^A,B,C Different capital letters in a column denote significant differences between fermented milk supplemented with different prebiotics (P < 0.05); ^a,b,c Different lowercase letters in a row for each fermented milk denote significant differences during storage period (P < 0.05). Abbreviations are: F: fermented milk without prebiotic addition including P1&P2, FC: fermented milk with carob powder including P1&P2, FI: fermented milk with inulin powder including P1&P2.
FP1	4.71 ± 0.10^aA	4.56 ± 0.02^abA	4.49 ± 0.07^bA	0.51 ± 0.12^aA	0.59 ± 0.05^abA	0.67 ± 0.10^bA	1,102 ± 0.08^aA	1,167 ± 0.15^aA	1,218 ± 0.16^bA
FP2	4.70 ± 0.04^aA	4.58 ± 0.10^bA	4.50 ± 0.04^bA	0.50 ± 0.08^aA	0.57 ± 0.10^aA	0.66 ± 0.13^aA	1,104 ± 0.06^aA	1,170 ± 0.16^aA	1,217 ± 0.12^bA
FP12	4.69 ± 0.03^aA	4.58 ± 0.11^aA	4.42 ± 0.11^bA	0.52 ± 0.04^aA	0.61 ± 0.12^aA	0.69 ± 0.08^aA	1,107 ± 0.12^aA	1,178 ± 0.12^aA	1,224 ± 0.08^bA
FCP1	4.57 ± 0.03^aB	4.46 ± 0.07^aB	4.35 ± 0.03^bB	0.69 ± 0.08^aB	0.80 ± 0.08^bB	0.96 ± 0.04^cB	1,336 ± 0.14^aB	1,368 ± 0.09^aB	1,408 ± 0.14^bB
FCP2	4.58 ± 0.04^aB	4.48 ± 0.05^aB	4.38 ± 0.03^bB	0.69 ±0.10^aB	0.79 ± 0.10^bB	0.91 ± 0.13^cB	1,324 ± 0.13^aB	1,360 ± 0.16^aB	1,398 ± 0.13^bB
FCP12	4.50 ± 0.03^aB	4.41 ± 0.03^aB	4.33 ± 0.10^bB	0.79 ± 0.06^aB	0.87 ± 0.12^bB	1.10 ± 0.20^cB	1,343 ± 0.12^aB	1,373 ± 0.15^aB	1,423 ± 0.14^bB
FIP1	4.60 ± 0.05^aC	4.52 ± 0.06^aA	4.41 ± 0.08^bA	0.56 ± 0.10^aA	0.76 ± 0.09^bB	0.83 ± 0.08^bC	1,203 ± 0.10^aC	1,271 ± 0.08^aC	1,301 ± 0.12^bC
FIP2	4.61 ± 0.04^aC	4.55 ± 0.12^aA	4.43 ± 0.07^bA	0.54 ± 0.10^aA	0.75 ± 0.05^bB	0.81 ± 0.05^bC	1,200 ± 0.16^aC	1,268 ± 0.14^aC	1,298 ± 0.08^bC
FP12	4.60 ± 0.04^aC	4.50 ± 0.10^aA	4.39 ± 0.02^bAB	0.60 ± 0.12^aA	0.78 ± 0.10^bB	0.89 ± 0.14^bC	1,218 ± 0.14^aC	1,276 ± 0.22^bC	1,324 ± 0.07^cC

Table 3. The changes of physico-chemical properties of different fermented milks during refrigerated storage*

Fermented milks

Titrable acidity (%)

Viscosity (cP)

Day 1

Day 14

Day 28

Day 1

Day 14

Day 28

Day 1

Day 14

Day 28

*Values were expressed as mean ± standard deviation (n = 3). ^A,B,C Different capital letters in a column denote significant differences between fermented milk supplemented with different prebiotics (P < 0.05); ^a,b,c Different lowercase letters in a row for each fermented milk denote significant differences during storage period (P < 0.05). Abbreviations are: F: fermented milk without prebiotic addition including P1&P2, FC: fermented milk with carob powder including P1&P2, FI: fermented milk with inulin powder including P1&P2.

FP1

4.71 ± 0.10^aA

4.56 ± 0.02^abA

4.49 ± 0.07^bA

0.51 ± 0.12^aA

0.59 ± 0.05^abA

0.67 ± 0.10^bA

1,102 ± 0.08^aA

1,167 ± 0.15^aA

1,218 ± 0.16^bA

FP2

4.70 ± 0.04^aA

4.58 ± 0.10^bA

4.50 ± 0.04^bA

0.50 ± 0.08^aA

0.57 ± 0.10^aA

0.66 ± 0.13^aA

1,104 ± 0.06^aA

1,170 ± 0.16^aA

1,217 ± 0.12^bA

FP12

4.69 ± 0.03^aA

4.58 ± 0.11^aA

4.42 ± 0.11^bA

0.52 ± 0.04^aA

0.61 ± 0.12^aA

0.69 ± 0.08^aA

1,107 ± 0.12^aA

1,178 ± 0.12^aA

1,224 ± 0.08^bA

FCP1

4.57 ± 0.03^aB

4.46 ± 0.07^aB

4.35 ± 0.03^bB

0.69 ± 0.08^aB

0.80 ± 0.08^bB

0.96 ± 0.04^cB

1,336 ± 0.14^aB

1,368 ± 0.09^aB

1,408 ± 0.14^bB

FCP2

4.58 ± 0.04^aB

4.48 ± 0.05^aB

4.38 ± 0.03^bB

0.69 ±0.10^aB

0.79 ± 0.10^bB

0.91 ± 0.13^cB

1,324 ± 0.13^aB

1,360 ± 0.16^aB

1,398 ± 0.13^bB

FCP12

4.50 ± 0.03^aB

4.41 ± 0.03^aB

4.33 ± 0.10^bB

0.79 ± 0.06^aB

0.87 ± 0.12^bB

1.10 ± 0.20^cB

1,343 ± 0.12^aB

1,373 ± 0.15^aB

1,423 ± 0.14^bB

FIP1

4.60 ± 0.05^aC

4.52 ± 0.06^aA

4.41 ± 0.08^bA

0.56 ± 0.10^aA

0.76 ± 0.09^bB

0.83 ± 0.08^bC

1,203 ± 0.10^aC

1,271 ± 0.08^aC

1,301 ± 0.12^bC

FIP2

4.61 ± 0.04^aC

4.55 ± 0.12^aA

4.43 ± 0.07^bA

0.54 ± 0.10^aA

0.75 ± 0.05^bB

0.81 ± 0.05^bC

1,200 ± 0.16^aC

1,268 ± 0.14^aC

1,298 ± 0.08^bC

FP12

4.60 ± 0.04^aC

4.50 ± 0.10^aA

4.39 ± 0.02^bAB

0.60 ± 0.12^aA

0.78 ± 0.10^bB

0.89 ± 0.14^bC

1,218 ± 0.14^aC

1,276 ± 0.22^bC

1,324 ± 0.07^cC

**Table 4.** Phenolic compounds of carob fermented milk (FCP12) during refrigerated storage (4 °C)*
Phenolic compounds	Carob powder	Day 0	Day 1	Day 14	Day 28
*Values are means of triplicates ± standard deviations (µg/g of fermented milk sample). Means followed by the different letters (a,b) in the same row represent a statistically significant difference (p < 0.05). Nd: not detected.
Gallic acid	162.50 ± 0.12	102.35 ± 0.14^a	100.82 ± 0.17^a	112.41 ± 0.07^a	78.06 ± 0.10^b
Proto-catechuic acid	39.19 ± 0.04	24.71 ± 0.04^a	21.20 ± 0.08^a	19.64 ± 0.11^a	10.27 ± 0.09^a
Chlorogenic acid	54.12 ± 0.11	33.67 ± 0.24^a	34.18 ± 0.20^a	35.21 ± 0.12^a	26.14 ± 0.07^a
Caffeic acid	Nd	Nd	Nd	Nd	Nd
p-coumaric acid	12.77 ± 0.11	8.11 ± 0.18^a	8.10 ± 0.12^a	11.04 ± 0.06^a	Nd
Ferulic acid	13.44 ± 0.12	7.47 ± 0.14^a	7.89 ± 0.08^a	10.41 ± 0.07^a	3.75 ± 0.14^b
o-coumaric acid	11.07 ± 0.04	6.33 ± 0.20^a	5.47 ± 0.11^a	4.15 ± 0.08^a	Nd
Trans- cinnamic acid	42.40 ± 0.15	24.10 ± 0.11^a	24.85 ± 0.22^a	18.47 ± 0.07^a	10.63 ± 0.15^b
Total phenolic acids (TPA)	335.49 ± 0.18	206.74 ± 0.14^a	202.51 ± 0.17^a	211.33 ± 0.17^a	128.85 ± 0.16^b
(+)-Catechin	11.02 ± 0.03	8.23 ± 0.10^a	8.21 ± 0.11^a	9.04 ± 0.11^a	Nd
Rutin	118.36 ± 0.02	88.76 ± 0.15^a	89.06 ± 0.08^a	89.86 ± 0.08^a	64.07 ± 0.20^b
Myricetin	18.17 ± 0.04	10.53 ± 0.11^a	10.50 ± 0.14^a	8.12 ± 0.15^ab	5.17 ± 0.08^b
Isoquercitrin	24.47 ± 0.06	15.20 ± 0.14^a	15.18 ± 0.15^a	15.89 ± 0.14^a	7.57 ± 0.11^b
Total flavonoids (TF)	172.02 ± 0.14	122.72 ± 0.16^a	122.95 ± 0.18^a	122.91 ± 0.17^a	76.81 ± 0.11^b
Total phenolics (TPA+TF)	507.51 ± 0.23	329.46 ± 0.20^a	325.46 ± 0.20^a	334.24 ± 0.20^a	205.66 ± 0.19^b

Table 4. Phenolic compounds of carob fermented milk (FCP12) during refrigerated storage (4 °C)*

Phenolic compounds

Carob powder

Day 0

Day 1

Day 14

Day 28

*Values are means of triplicates ± standard deviations (µg/g of fermented milk sample). Means followed by the different letters (a,b) in the same row represent a statistically significant difference (p < 0.05). Nd: not detected.

Gallic acid

162.50 ± 0.12

102.35 ± 0.14^a

100.82 ± 0.17^a

112.41 ± 0.07^a

78.06 ± 0.10^b

Proto-catechuic acid

39.19 ± 0.04

24.71 ± 0.04^a

21.20 ± 0.08^a

19.64 ± 0.11^a

10.27 ± 0.09^a

Chlorogenic acid

54.12 ± 0.11

33.67 ± 0.24^a

34.18 ± 0.20^a

35.21 ± 0.12^a

26.14 ± 0.07^a

Caffeic acid

p-coumaric acid

12.77 ± 0.11

8.11 ± 0.18^a

8.10 ± 0.12^a

11.04 ± 0.06^a

Ferulic acid

13.44 ± 0.12

7.47 ± 0.14^a

7.89 ± 0.08^a

10.41 ± 0.07^a

3.75 ± 0.14^b

o-coumaric acid

11.07 ± 0.04

6.33 ± 0.20^a

5.47 ± 0.11^a

4.15 ± 0.08^a

Trans- cinnamic acid

42.40 ± 0.15

24.10 ± 0.11^a

24.85 ± 0.22^a

18.47 ± 0.07^a

10.63 ± 0.15^b

Total phenolic acids (TPA)

335.49 ± 0.18

206.74 ± 0.14^a

202.51 ± 0.17^a

211.33 ± 0.17^a

128.85 ± 0.16^b

(+)-Catechin

11.02 ± 0.03

8.23 ± 0.10^a

8.21 ± 0.11^a

9.04 ± 0.11^a

Rutin

118.36 ± 0.02

88.76 ± 0.15^a

89.06 ± 0.08^a

89.86 ± 0.08^a

64.07 ± 0.20^b

Myricetin

18.17 ± 0.04

10.53 ± 0.11^a

10.50 ± 0.14^a

8.12 ± 0.15^ab

5.17 ± 0.08^b

Isoquercitrin

24.47 ± 0.06

15.20 ± 0.14^a

15.18 ± 0.15^a

15.89 ± 0.14^a

7.57 ± 0.11^b

Total flavonoids (TF)

172.02 ± 0.14

122.72 ± 0.16^a

122.95 ± 0.18^a

122.91 ± 0.17^a

76.81 ± 0.11^b

Total phenolics (TPA+TF)

507.51 ± 0.23

329.46 ± 0.20^a

325.46 ± 0.20^a

334.24 ± 0.20^a

205.66 ± 0.19^b

**Table 5.** Phenolic compounds of carob fermented milk (FCP12) during *in vitro* digestion*
Phenolic compounds	Carob powder	BF	Oral	Gastric	Intestinal	Bioaccessibility (%)
*Results are expressed as μg of individual compound in 1 g of carob fermented milk. Means followed by the different letters (a,b) in the same row represent a statistically significant difference (p < 0.05). BF: Before digestion. Nd: not detected/determined?
Gallic acid	162.50 ± 0.12	102.35 ± 0.14^a	104.56 ± 1.02^a	234.04 ± 1.03^b	452.12 ± 0.99^c	441.74
Proto-catechuic acid	39.19 ± 0.04	24.71 ± 0.04^a	24.53 ± 1.02^a	44.76 ± 0.99^b	65.49 ± 1.05^c	265.03
Chlorogenic acid	54.12 ± 0.11	33.67 ± 0.24^a	36.78 ± 0.05^a	78.23 ± 1.02^b	102.76 ± 0.75^c	305.19
Caffeic acid	Nd	Nd	Nd	Nd	6.45 ± 0.08^a	Nd
p-coumaric acid	12.77 ± 0.11	8.11 ± 0.18^a	10.02 ± 0.08^a	16.06 ± 0.45^a	21.12 ± 0.98^a	260.41
Ferulic acid	13.44 ± 0.12	7.47 ± 0.14^a	8.45 ± 0.99^a	15.17 ± 0.87^b	10.23 ± 0.07^a	136.94
o-coumaric acid	11.07 ± 0.04	6.33 ± 0.20^a	5.33 ± 0.05^a	Nd	Nd	Nd
Trans- cinnamic acid	42.40 ± 0.15	24.10 ± 0.11^a	26.12 ± 0.08^a	56.58 ± 0.99^b	67.44 ± 1.05^b	279.83
Total phenolic acids (TPA)	335.46 ± 0.21	206.74 ± 0.15^a	215.79 ± 0.17^a	444.84 ± 0.21^b	725.61 ± 0.24^c	Nd
(+)-Catechin	11.02 ± 0.03	8.23 ± 0.10^a	7.49 ± 0.08^a	26.19 ± 0.08^b	40.02 ± 1.05^c	486.26
Rutin	118.36 ± 0.02	88.76 ± 0.15^a	90.02 ± 0.10^a	128.73 ± 0.88^b	264.14 ± 1.10^c	297.58
Myricetin	18.17 ± 0.04	10.53 ± 0.11^a	12.37 ± 0.08^a	28.21 ± 0.09^b	35.12 ± 1.00^b	333.52
Isoquercitrin	24.47 ± 0.06	15.20 ± 0.14^a	16.75 ± 0.66^a	8.12 ± 0.06^b	5.36 ± 0.09^b	35.26
Total flavonoids (TF)	172.02 ± 0.14	122.72 ± 0.10^a	126.63 ± 0.11^a	191.25 ± 0.16^b	344.64 ± 0.14^c	Nd
Total phenolics (TPA+TF)	510.88 ± 0.23	329.46 ± 0.17^a	342.42 ± 0.16^a	636.09 ± 0.20^b	1,070.25 ± 0.25^c	Nd

Table 5. Phenolic compounds of carob fermented milk (FCP12) during in vitro digestion*

Phenolic compounds

Carob powder

Oral

Gastric

Intestinal

Bioaccessibility (%)

*Results are expressed as μg of individual compound in 1 g of carob fermented milk. Means followed by the different letters (a,b) in the same row represent a statistically significant difference (p < 0.05). BF: Before digestion. Nd: not detected/determined?

Gallic acid

162.50 ± 0.12

102.35 ± 0.14^a

104.56 ± 1.02^a

234.04 ± 1.03^b

452.12 ± 0.99^c

441.74

Proto-catechuic acid

39.19 ± 0.04

24.71 ± 0.04^a

24.53 ± 1.02^a

44.76 ± 0.99^b

65.49 ± 1.05^c

265.03

Chlorogenic acid

54.12 ± 0.11

33.67 ± 0.24^a

36.78 ± 0.05^a

78.23 ± 1.02^b

102.76 ± 0.75^c

305.19

Caffeic acid

6.45 ± 0.08^a

p-coumaric acid

12.77 ± 0.11

8.11 ± 0.18^a

10.02 ± 0.08^a

16.06 ± 0.45^a

21.12 ± 0.98^a

260.41

Ferulic acid

13.44 ± 0.12

7.47 ± 0.14^a

8.45 ± 0.99^a

15.17 ± 0.87^b

10.23 ± 0.07^a

136.94

o-coumaric acid

11.07 ± 0.04

6.33 ± 0.20^a

5.33 ± 0.05^a

Trans- cinnamic acid

42.40 ± 0.15

24.10 ± 0.11^a

26.12 ± 0.08^a

56.58 ± 0.99^b

67.44 ± 1.05^b

279.83

Total phenolic acids (TPA)

335.46 ± 0.21

206.74 ± 0.15^a

215.79 ± 0.17^a

444.84 ± 0.21^b

725.61 ± 0.24^c

(+)-Catechin

11.02 ± 0.03

8.23 ± 0.10^a

7.49 ± 0.08^a

26.19 ± 0.08^b

40.02 ± 1.05^c

486.26

Rutin

118.36 ± 0.02

88.76 ± 0.15^a

90.02 ± 0.10^a

128.73 ± 0.88^b

264.14 ± 1.10^c

297.58

Myricetin

18.17 ± 0.04

10.53 ± 0.11^a

12.37 ± 0.08^a

28.21 ± 0.09^b

35.12 ± 1.00^b

333.52

Isoquercitrin

24.47 ± 0.06

15.20 ± 0.14^a

16.75 ± 0.66^a

8.12 ± 0.06^b

5.36 ± 0.09^b

35.26

Total flavonoids (TF)

172.02 ± 0.14

122.72 ± 0.10^a

126.63 ± 0.11^a

191.25 ± 0.16^b

344.64 ± 0.14^c

Total phenolics (TPA+TF)

510.88 ± 0.23

329.46 ± 0.17^a

342.42 ± 0.16^a

636.09 ± 0.20^b

1,070.25 ± 0.25^c

**Table 6.** Antibacterial activity in the fermented milk supplemented with 4% carob powder*
	Inhibitory zone diameter
*Values are represented as mean ± SD (n=3). Means followed by the different letters (a,b) in the same row indicate a statistically significant difference (p < 0.05).
Control	11 ± 0.2^a	9 ± 0.3^a	10 ± 0.2^a	10 ± 0.3^a
Fermented milk (FCP12)	12 ± 0.4^b	14 ± 0.2^b	11 ± 0.1^b	13 ± 0.2^b

Table 6. Antibacterial activity in the fermented milk supplemented with 4% carob powder*

Inhibitory zone diameter

L. monocytogenes

S. aureus

S. typhimurium

E. coli

*Values are represented as mean ± SD (n=3). Means followed by the different letters (a,b) in the same row indicate a statistically significant difference (p < 0.05).

Control

11 ± 0.2^a

9 ± 0.3^a

10 ± 0.2^a

10 ± 0.3^a

Fermented milk (FCP12)

12 ± 0.4^b

14 ± 0.2^b

11 ± 0.1^b

13 ± 0.2^b