Characterization of plant-based meat alternatives blended with anthocyanins, chlorophyll, and various edible natural pigments

Figures & data

Figure 1. Pigments extracted from natural sources. 30ml of all samples were prepared in a 50ml conical tube (Φ 3cm) each. The absorbance at 535 nm of all of them is adjusted to 0.700.

S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB, S15 = Sample Anthocyanin (Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control)

Table 1. Composition of PBM.

Ingredient	Content (%)	Remarks
TVP	54.45–52.75	TVP imitates the fibrillar structure of meat muscle and major extender
Shiitake mushrooms	8.00	Umami flavor potentiators and flavor enhancers
ISP	4.50	Act as an emulsifying agent and has elastic textural abilities like meat
IWP	4.50	Increased bulk density, emulsification and textural abilities
Tapioca starch	2.50	Thickener, stabilizer and texturizing agent
Fats	8.00	Taste enhancer, texture, appearance and aroma
Salt	0.20	Overall flavor
Seasoning	2.00	Masking cereal notes, enhance flavor and odour
MC	2.00	Binder gelling and stabilizer
Garlic powder	1.00	Flavour enhancer
Molasses	2.80	Textural, soft and chewy sensation
Natural pigments	0.05–1.75	Imitate the myoglobin color
Ice	10.00	Esthetic and organoleptic properties

Purified water 3 times of TVP for rehydration

TVP; Textured vegetable protein, ISP; isolate soy protein, IWP; isolate wheat protein, MC; Methylcellulose

Figure 2. Reaction scheme for the synthesis of pheophytin and Fe-pheophytin. Derivatives from natural chlorophyll are present in crude spinach extract (PubChem).

Figure 3. Flow diagram of PBM incorporated with natural pigments.

Table 2. Pigment weight and concentration per colorant.

Sample	Concentrations (%)	Pigments weight (g)
Dilute red 1	0.13	.04
Dilute red 2	0.13	.04
Red color CG2	0.22	.065
Paprika	0.05	.015
Monascus Color No.30	0.08	.025
Red RR	0.10	.03
Purple Grape	0.17	.05
Cherry Red	0.28	.08
Monascus Color 100	0.18	.06
Red Cabbage (liquid)	0.25	.08
Red Cabbage 100	0.32	.10
AF Beet Red 30	0.40	.12
Grape Skin Color	0.15	.04
Red Color PB	0.37	.11
Sample anthocyanin	0.54	.18
Sample Fe-chlorophyll	1.00	.30
Myoglobin (control)	1.76	.53

Each pigment contained 30 mL in volume.

Figure 4. Visible appearance of raw PBM patties incorporated with natural pigments.

Figure 5. Visible appearance of steamed PBM patties incorporated with natural pigments. S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB, S15 = Sample Anthocyanin (Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control).

Figure 6. Visible appearance of cooked PBM patties incorporated with natural pigments.

S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB, S15 = Sample Anthocyanin (Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control)

Table 3. Color coordinates of raw, steamed, and cooked patties incorporated with natural pigments.

Samples	Color (Raw patties)			Color (Steam patties)			Color (Cooked patties)
	L*	a*	b*	L*	a*	b*	L*	a*	b*
S 1	53.12 ± 0.67 ^abc	11.93 ± 1.35 ^d	16.19 ± 0.68 ^ef	42.39 ± 0.42 ^a	17.26 ± 2.61 ^b	17.72 ± 0.27 ^de	42.98 ± 1.50 ^ab	17.75 ± 0.63 ^ab	18.56 ± 0.60 ^def
S 2	48.35 ± 0.35 ^ef	16.83 ± 0.56 ^b	17.06 ± 0.45 ^de	36.56 ± 1.54 ^cde	16.16 ± 0.86 ^b	17.54 ± 0.55 ^de	39.83 ± 0.60 ^abcde	16.62 ± 0.79 ^b	20.27 ± 0.99 ^bcde
S 3	50.45 ± 1.99 ^cde	12.03 ± 0.51 ^d	13.60 ± 0.65 ^h	38.31 ± 1.55 ^abcde	13.81 ± 0.55 ^c	15.29 ± 0.61 ^ef	38.92 ± 0.26 ^abcde	15.27 ± 0.38 ^b	15.93 ± 0.14 ^fgh
S 4	53.81 ± 0.48 ^ab	9.40 ± 0.28 ^e	27.92 ± 0.48 ^a	40.67 ± 0.76 ^abc	11.16 ± 0.08 ^d	25.24 ± 0.90 ^a	43.15 ± 0.85 ^ab	10.42 ± 0.45 ^cde	26.05 ± 1.39 ^a
S 5	52.87 ± 1.94 ^abcd	9.10 ± 0.70 ^e	21.45 ± 0.40 ^bc	38.85 ± 2.94 ^abcde	11.88 ± 0.94 ^cd	20.16 ± 1.68 ^bcd	43.93 ± 0.82 ^a	11.41 ± 1.21 ^c	21.48 ± 1.31 ^bcd
S 6	52.86 ± 0.75 ^abcd	7.08 ± 0.36 ^f	15.78 ± 0.67 ^efg	40.31 ± 1.21 ^abcd	7.66 ± 0.11 ^ef	18.17 ± 0.19 ^cde	41.62 ± 2.44 ^abcd	8.67 ± 0.60 ^defg	18.59 ± 2.11 ^def
S 7	46.56 ± 1.33 ^fg	1.75 ± 0.06 ^j	8.37 ± 0.78 ^lm	35.79 ± 2.10 ^de	4.42 ± 0.20 ^g	9.75 ± 1.48 ^h	36.58 ± 0.86 ^defg	7.03 ± 1.31 ^gh	12.97 ± 1.17 ^hij
S 8	45.82 ± 0.36 ^fg	3.22 ± 0.04 ^i	9.66 ± 0.96 ^jkl	34.49 ± 2.03 ^e	5.23 ± 0.03 ^fg	10.93 ± 0.97 ^gh	36.15 ± 2.95 ^efg	4.69 ± 0.63 ^h	9.18 ± 1.25 ^jk
S 9	33.84 ± 0.56 ^j	25.15 ± 0.39 ^a	14.75 ± 0.29 ^fgh	29.02 ± 0.75 ^f	20.63 ± 0.85 ^a	11.43 ± 1.18 ^gh	28.28 ± 0.16 ^h	19.26 ± 1.60 ^a	9.51 ± 0.92 ^jk
S 10	49.84 ± 0.58 ^de	3.06 ± 0.08 ^ij	10.31 ± 0.09 ^ijk	41.62 ± 1.93 ^ab	6.19 ± 0.11 ^efg	15.95 ± 1.42 ^ef	38.77 ± 1.07 ^bcde	7.23 ± 0.19 ^fg	15.19 ± 0.18 ^fgh
S 11	44.55 ± 1.01 ^g	5.15 ± 0.25 ^gh	7.51 ± 0.50 ^m	37.19 ± 0.98 ^bcde	7.47 ± 0.18 ^ef	13.29 ± 0.44 ^fg	40.84 ± 1.42 ^abcde	10.66 ± 0.17 ^cd	16.93 ± 0.97 ^efg
S 12	50.34 ± 0.22 ^cde	13.31 ± 0.07 ^c	14.42 ± 0.03 ^gh	42.13 ± 0.28 ^a	8.27 ± 0.03 ^e	21.07 ± 0.13 ^b	42.85 ± 1.84 ^abc	9.35 ± 0.36 ^cdefg	23.80 ± 1.15 ^ab
S 13	46.38 ± 0.44 ^fg	2.89 ± 0.09 ^ij	9.00 ± 0.34 ^klm	36.06 ± 0.70 ^cde	5.60 ± 0.03 ^fg	11.56 ± 0.61 ^gh	37.85 ± 1.32 ^cdef	8.77 ± 0.10 ^cdefg	14.23 ± 0.07 ^ghi
S 14	41.01 ± 0.59 ^h	6.15 ± 0.08 ^fg	3.93 ± 0.62 ^n	29.67 ± 0.81 ^f	7.73 ± 0.35 ^ef	4.42 ± 0.60 ^i	33.54 ± 2.04 ^fg	9.90 ± 0.94 ^cdef	8.91 ± 2.23 ^k
S 15	47.79 ± 0.82 ^ef	2.51 ± 0.23 ^ij	11.51 ± 0.53 ^i	35.08 ± 1.16 ^e	5.92 ± 0.25 ^efg	11.60 ± 1.37 ^gh	39.59 ± 0.45 ^abcde	8.43 ± 0.36 ^defg	18.00 ± 0.62 ^defg
S 16	49.94 ± 1.21 ^de	6.54 ± 0.09 ^f	18.24 ± 0.54 ^d	37.07 ± 1.46 ^bcde	7.64 ± 0.26 ^ef	16.41 ± 0.96 ^e	32.45 ± 1.13 ^gh	8.34 ± 0.88 ^defg	11.03 ± 0.43 ^ijk
S 17	51.77 ± 1.17 ^bcd	7.06 ± 0.14 ^fg	21.81 ± 0.41 ^b	41.66 ± 0.89 ^ab	5.41 ± 0.27 ^fg	18.04 ± 0.72 ^cde	39.39 ± 2.46 ^abcde	7.82 ± 0.88 ^efg	19.02 ± 2.13 ^cdef
S 18	38.10 ± 0.78 ^i	23.77 ± 0.41 ^b	11.32 ± 0.67 ^ij	42.66 ± 1.27 ^a	6.29 ± 0.44 ^efg	20.66 ± 0.32 ^bc	41.05 ± 1.26 ^abcde	10.20 ± 1.29 ^cde	22.50 ± 1.07 ^abc

The results are shown as mean ± standard deviation (n = 3). a, b, c, d Means with the different letters are significantly different (p < .05). S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB, S15 = Sample Anthocyanin (Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control), S18= Negative control (beef patty)

Table 4. Proximate chemical composition of PBM incorporated with natural pigments.

	Treatment
Samples	Moisture Content (%)	Crude Protein (%)	Crude Fat (%)	Ash (%)
S 1	62.27 ± 0.34^ab	19.24 ± 0.56bc	3.19 ± 0.64^ab	2.14 ± 0.04^bc
S 2	61.63 ± 0.37^ab	2.98 ± 0.52ab	3.16 ± 0.48 ^ab	2.05 ± 0.02^bc
S 3	59.23 ± 0.35^abc	19.78 ± 0.42bc	3.06 ± 0.18 ^ab	2.16 ± 0.06^bc
S 4	61.25 ± 0.36^ab	21.01 ± 0.56ab	3.09 ± 0.17 ^aa	2.26 ± 0.05^bc
S 5	63.28 ± 0.31^ab	2.12±.53 ab	3.17 ± 1.00 ^ab	2.16 ± 0.01^ab
S 6	63.39 ± 0.25^ab	19.45 ± 0.40ab	3.02 ± 0.19 ^ab	1.96 ± 0.16^ab
S 7	62.78 ± 0.33^ab	2.02 ± 0.50ab	2.99 ± 0.54^abc	2.22 ± 0.08^ab
S 8	63.98 ± 0.29^a	19.78 ± 0.41ab	3.14 ± 0.29^ab	1.99 ± 0.10^ab
S 9	64.35 ± 0.25^ab	2.06 ± 0.51ab	2.95 ± 0.53^ab	2.19 ± 0.10^ab
S 10	63.28 ± 0.40^abc	2.05 ± 052ab	3.16 ± 0.25^ab	2.14 ± 0.05 a^b
S 11	62.45 ± 0.32^ab	19.77 ± 045ab	2.92 ± 0.43^abv	2.26 ± 0.10^ab
S 12	63.69 ± 0.24^ab	19.86 ± 0.51ab	2.99 ± 0.44^abv	2.15 ± 0.09^ab
S 13	62.12 ± 0.33^ab	2.09 ± 0.53ab	3.20 ± 0.23^ab	1.96 ± 0.01^ab
S 14	62.27 ± 0.34 ^abc	19.17±.69 ab	3.16 ± 0.15 ^a	2.25 ± 0.04 ^ab
S 15	61.92 ± 0.36 ^abc	19.32±.41 ab	2.93 ± 0.40 ^ab	3.80 ± 0.05 ^ac
S 16	62.31 ± 0.26 ^ab	19.28±.24 ab	3.08 ± 0.18 ^ab	3.25 ± 0.04 ^ac
S 17	61.18 ± 0.13 ^ab	2.50±.54 abc	3.18 ± 0.21 ^ab	1.95 ± 0.05 ^b
S18	62.27 ± 0.34 ^abc	19.17±.69 ab	3.16 ± 0.15 ^ab	1.71 ± 0.03 ^d

The results are shown as mean ± standard deviation (n = 3). a, b, c, d Means with the different letters are significantly different (p < .05). S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB S15 = Sample Anthocyanin (Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control), S18 = Negative control (beef patty)

Figure 7. Absorbance measurement results of pigment samples.

S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB, S15 = Sample Anthocyanin(Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control)

Table 5. Texture Profile Analysis (TPA) of PBM incorporated with natural pigments.

Samples	Texture Profile
	Chewiness (mJ)	Hardness (N)	Gumminess (N)	Springiness (mm)	Cohesiveness (%)	Resilience (%)
S 1	7.43 ± 0.43 ^bcd	5.11 ± 0.85 ^bcde	2.44 ± 0.17 ^bcde	3.06 ± 0.07 ^ab	0.46 ± 0.03 ^abc	0.18 ± 0.02 ^abc
S 2	7.67 ± 1.05 ^bcd	5.27 ± 0.53 ^bcde	2.62 ± 0.26 ^bcd	2.91 ± 0.11 ^bc	0.50 ± 0.00 ^a	0.21 ± 0.01 ^a
S 3	8.03 ± 0.84 ^bcd	5.95 ± 0.38 ^bcd	2.74 ± 0.17 ^bcd	2.92 ± 0.12 ^bc	0.46 ± 0.01 ^abc	0.18 ± 0.01 ^abc
S 4	9.27 ± 0.15 ^b	6.63 ± 0.36 ^bc	3.04 ± 0.06 ^b	3.06 ± 0.05 ^ab	0.46 ± 0.02 ^abc	0.19 ± 0.02 ^abc
S 5	8.77 ± 0.62 ^bc	6.93 ± 0.47 ^b	3.04 ± 0.19 ^b	2.88 ± 0.04 ^bc	0.44 ± 0.00 ^abc	0.17 ± 0.00 ^abc
S 6	8.07 ± 0.38 ^bcd	6.11 ± 0.17 ^bcd	2.77 ± 0.11 ^bc	2.91 ± 0.01 ^bc	0.45 ± 0.01 ^abc	0.18 ± 0.00 ^abc
S 7	6.93 ± 0.84 ^bcde	5.60 ± 0.40 ^bcde	2.48 ± 0.22 ^bcd	2.78 ± 0.09 ^bc	0.44 ± 0.01 ^abc	0.18 ± 0.01 ^abc
S 8	6.00 ± 0.50 ^cde	5.48 ± 0.39 ^bcde	2.23 ± 0.18 ^bcde	2.69 ± 0.02 ^c	0.40 ± 0.00 ^c	0.16 ± 0.00 ^c
S 9	6.57 ± 0.44 ^bcde	5.34 ± 0.39 ^bcde	2.33 ± 0.06 ^bcde	2.82 ± 0.15 ^bc	0.44 ± 0.03 ^abc	0.16 ± 0.02 ^bc
S 10	5.83 ± 1.29 ^cde	4.45 ± 0.86 ^de	2.13 ± 0.39 ^cde	2.70 ± 0.15 ^c	0.48 ± 0.01 ^ab	0.20 ± 0.01 ^ab
S 11	5.27 ± 0.90 ^de	4.39 ± 0.63 ^de	2.04 ± 0.36 ^cde	2.60 ± 0.08 ^c	0.46 ± 0.03 ^abc	0.18 ± 0.02 ^abc
S 12	6.47 ± 0.64 ^bcde	4.99 ± 0.29 ^cde	2.29 ± 0.13 ^bcde	2.81 ± 0.13 ^bc	0.46 ± 0.02 ^abc	0.18 ± 0.02 ^abc
S 13	5.77 ± 0.32 ^de	4.72 ± 0.66 ^de	2.09 ± 0.17 ^cde	2.78 ± 0.10 ^bc	0.45 ± 0.03 ^abc	0.19 ± 0.01 ^abc
S 14	7.23 ± 0.78 ^bcd	5.91 ± 0.41 ^bcd	2.52 ± 0.23 ^bcd	2.86 ± 0.04 ^bc	0.42 ± 0.01 ^bc	0.16 ± 0.00 ^bc
S 15	4.13 ± 0.64 ^e	3.72 ± 0.48 ^e	1.60 ± 0.18 ^e	2.57 ± 0.17 ^c	0.43 ± 0.02 ^abc	0.18 ± 0.01 ^abc
S 16	5.93 ± 1.98 ^cde	4.51 ± 1.00 ^de	2.11 ± 0.58 ^cde	2.70 ± 0.18 ^c	0.46 ± 0.03 ^abc	0.19 ± 0.02 ^abc
S 17	16.87 ± 1.13 ^a	11.23 ± 0.26 ^a	5.09 ± 0.28 ^a	3.31 ± 0.05 ^a	0.45 ± 0.04 ^abc	0.19 ± 0.01 ^abc
S 18	5.10 ± 0.87 ^de	4.54 ± 0.64 ^de	1.89 ± 0.25 ^de	2.68 ± 0.12 ^c	0.42 ± 0.01 ^bc	0.17 ± 0.01 ^bc

The results are shown as mean ± standard deviation (n = 3). a, b, c, d Means with the different letters are significantly different (p < .05). S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB S15 = Sample Anthocyanin (Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control), S18 = Negative control

Figure 8. DPPH radical scavenging activity of PBM incorporated with natural pigments.

Sample = S S1 = Dilute Red 1, S2 = Dilute Red 2, S3 = Red color CG2, S4 = Paprika, S5 = Monascus Color No.30, S6 = Red RR, S7 = Purple Grape, S8 = Cherry Red, S9 = Monascus Color 100, S10 = Red Cabbage (liquid), S11 = Red Cabbage 100, S12 = AF Beet Red 30, S13 = Grape skin Color, S14 = Red Color PB, S15 = Sample Anthocyanin(Sample A), S16 = Sample Fe-pheophytin, S17 = Myoglobin (control)

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article. Find additional templates here: Writing a data availability statement – Author Services (taylorandfrancis.com). Access to raw data is possible upon justifiable request.