ANTHOCYANIN-BASED COLORANT

Abstract

1. A composition, which is an anthocyanin-based colorant composition comprising 50-90 mol-%, based on the total amount of anthocyanins, of pelargonidin-based anthocyanins, and wherein (i) >70 mol-% of all anthocyanins are acylated with at least one phenolic acid; and (ii) >20 mol-% of all anthocyanins are acylated with at least one hydroxycinnamic acid; and wherein the composition has a red color with a hue value H in the L*C*h* color system in the range of 10-30, measured at an L*-value of (70.00.1) in a 0.1 mol/l trisodium citrate dihydrate buffer at pH 3 in a 1 cm-length quartz cell using Spectraflash 650 (Datacolor) in transmission mode under D65 illuminant 10 Deg. Also, the present invention relates to use of the above compositions as a food colorant.

Claims

1-15. (canceled)

16. A process for obtaining an anthocyanin-based food colorant composition from red Ipomoea batatas sweet potatoes, comprising: (a) washing red variety Ipomoea batatas sweet potato tubers or a juice or extract thereof with acidified water to obtain an acidified extract comprising anthocyanins; and (b) filtering the acidified extract to obtain a food colorant composition comprising anthocyanins extracted from the sweet potatoes, wherein 50-90 mol % of the extracted anthocyanins present in the composition are pelargonidin-based anthocyanins extracted from the sweet potatoes, wherein (i) 70 mol-% of the extracted anthocyanins present in the composition are acylated with at least one phenolic acid; and (ii) 20 mol % of the extracted anthocyanins present in the composition are acylated with at least one hydroxycinnamic acid; and wherein the food colorant composition has a red color with a hue value H in the L*C*h* color system in the range of 10-30, measured at an L*-value of (70.00.1) in a 0.1 mol/L trisodium citrate dihydrate buffer at pH 3 in a 1 cm-length quartz cell using Spectraflash 650 (Datacolor) in transmission mode under D65 illuminant 10 Deg.

17. The process of claim 16, wherein the food colorant composition has a red color with a hue value H in the L*C*h* color system in the range of 13-27.

18. The process of claim 16, wherein the food colorant composition contains no or only trace amounts of sulfur compounds.

19. The process of claim 16, wherein 4-15 mol % of the extracted anthocyanins present in the food colorant composition are peonidin-based anthocyanins.

20. The process of claim 16, wherein 80 mol % of the extracted anthocyanins present in the food colorant composition are acylated with at least one phenolic acid.

21. The process of claim 16, wherein up to 80 mol % of the extracted anthocyanins present in the food colorant composition are acylated with at least one hydroxycinnamic acid.

22. The process of claim 16, wherein 5-55 mol % of the extracted anthocyanins present in the food colorant composition is an acylated pelargonidin derivative of formula (1), shown in the protonized, positively charged form: ##STR00008##

23. The process of claim 16, wherein 3-60 mol % of the extracted anthocyanins present in the food colorant composition is an acylated pelargonidin derivative of formula (2), shown in the protonized, positively charged form: ##STR00009##

24. The process of claim 16, wherein the food colorant composition has an anthocyanin content, in terms of kuromanin equivalents, of 2-30 mg/ml for a colorant at 40-60% dry matter.

25. The process of claim 16, wherein step (a) comprises washing red variety Ipomoea batatas sweet potato tubers with acidified water.

26. The process of claim 16, further comprising removing other anthocyanins from the food colorant composition obtained at step (b).

27. The process of claim 16, wherein the process does not include supplementing the food colorant composition obtained at step (b) with additional anthocyanins.

28. The process of claim 16, further comprising purifying the food colorant composition obtained at step (b).

29. The method of claim 16, further comprising pasteurizing the food colorant composition obtained at step (b).

30. The process of claim 16, further comprising concentrating the food colorant composition obtained at step (b) to obtain a concentrate.

31. The process of claim 40, wherein the concentrate has a dry matter content of 10-95 wt. %.

32. A process for coloring a food, comprising: (a) washing red variety Ipomoea batatas sweet potato tubers or a juice or extract thereof with acidified water to obtain an acidified extract comprising anthocyanins; (b) filtering the acidified extract to obtain a food colorant composition comprising anthocyanins extracted from the sweet potatoes, wherein 50-90 mol % of the extracted anthocyanins present in the composition are pelargonidin-based anthocyanins extracted from the sweet potatoes, wherein (i) 70 mol-% of the extracted anthocyanins present in the composition are acylated with at least one phenolic acid; and (ii) 20 mol % of the extracted anthocyanins present in the composition are acylated with at least one hydroxycinnamic acid; and wherein the food colorant composition has a red color with a hue value H in the L*C*h* color system in the range of 10-30, measured at an L*-value of (70.00.1) in a 0.1 mol/L trisodium citrate dihydrate buffer at pH 3 in a 1 cm-length quartz cell using Spectraflash 650 (Datacolor) in transmission mode under D65 illuminant 10 Deg; and (c) adding the food colorant composition to a food.

33. The process of claim 32, wherein the food is a beverage, fruit preparation, dairy product, ice cream, or confectionary.

34. The process of claim 32, wherein the food is a beverage.

35. An anthocyanin-based food colorant composition obtained by a process according to claim 16.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0061] FIG. 1 Chromatographic profile (top) at 520 nm and mass fingerprint (bottom) of anthocyanins from red sweet potato.

[0062] FIG. 2 UV-visible spectra of the two major anthocyanins isolated from red sweet potato extract.

[0063] FIG. 3 Structure of Anthocyanin 1.

[0064] FIG. 4 Structure of Anthocyanin 2.

[0065] FIG. 5 Evolution of DE* 2000 during the 2-month storage of colored model beverage medium in cold room.

[0066] FIG. 6 Evolution of DE* 2000 during the 2-month storage of colored model beverage medium under light exposure.

[0067] FIG. 7 Evolution of DE* 2000 during the 2-month storage of colored model beverage medium under heat exposure.

[0068] FIG. 8 Evolution of DE* 2000 during the 2-month storage of colored model beverage medium containing ascorbic acid in cold room.

[0069] FIG. 9 Evolution of DE* 2000 during the 2-month storage of colored model beverage medium containing ascorbic acid under light exposure.

[0070] FIG. 10 Evolution of DE* 2000 during the 2-month storage of colored model beverage medium containing ascorbic acid under heat exposure.

[0071] FIG. 11 Evolution of DE* 2000 during the 2-week storage of fruit preparation/white mass blends in cold room at 4 C.

[0072] FIG. 12 Evolution of DE* 2000 during the 1-month storage of the colored fruit preparation.

EXAMPLES

[0073] As an illustrative example of a colorant of the invention and a procedure for its preparation the extraction and characterization of anthocyanins from Red Sweet Potato (RSWP), i.e. the red variety of Ipomoea batatas (L.) Lam, will be described.

[0074] As a reference, carminic acid has been used in some instances herein, since it obtains a very stable orange shade when it is added to beverages and thus can be seen as a reference colorant in at least this field of application. However, the legally admissible concentrations of carminic acid in beverages are too low to achieve the color shades as achievable with anthocyanins, and especially as achievable with the composition of the present invention. Thus, while a comparison with carminic acid at the intended color shades might show a superior stability of carminic acid, such an embodiment would be unsuitable in practice due to inadmissible high carminic acid levels. Therefore, carminic acid has been used as a reference only, for comparison known anthocyanins have been chosen.

Example 1

[0075] Characterization of Major Anthocyanins from RSWP

A) Extraction

[0076] A RSWP concentrate prepared by chopping RSWP tubers into slices and washing them 4 times with acidified water to conduct an extraction, and then subjecting the obtained extract to micro-filtration and subsequent purified on an absorbing resin. The resulting anthocyanin extract was concentrated and then pasteurized.

[0077] Anthocyanins were isolated and concentrated on a Sep-Pak C18 cartridge (Waters)) The cartridge was washed with 2 ml of methanol and then with 2 ml of acidified water (HCl 0.3%) before loading few drops of RSWP concentrate. Once the cartridge has been washed with 4 ml of acidified water (HCl 0.3%), and then with 2 ml of ethyl acetate, anthocyanins were finally eluted with a minimum volume of acidified methanol (HCl 0.1%).

B) Analysis

[0078] The anthocyanin extract was analyzed by fast-HPLC/ESI-TI. Separation was obtained at 30 C. using a BEH (Waters) C18 Column (50 mm2.1 mm, 1.8 m), by injecting 1 L of the filtered extract. The mobile phase consisted of two solvents: A, Water/Acetonitrile/HCOOH (95.7/3.3/1, v/v/v) and B, Water/Acetonitrile/HCOOH (44/55/1, v/v/v) at a flow rate of 0.8 ml/min. The gradient used was as follows:

TABLE-US-00001 Time Solvent A Solvent B in min (vol.-%) (vol.-%) 0 94 6 3.3 80 20 6 60 40 7 40 60 8 94 6 11 94 6

[0079] Mass spectrometry analyses were performed on a Bruker Daltonics HCT Ultra, operating in the positive electrospray ionization mode. Ions to be fragmented in MS.sup.2 or MS.sup.3 were automatically chosen by the software. Fragmentation is obtained through a screening of power leading in a determination of the minimal power necessary to break down the linkages.

C) Results and Discussion

[0080] The chromatographic profile of anthocyanins from RSWP is presented on FIG. 1. This profile reflects the presence of two major peaks, eluted at 28.8 and 37.6 min, and numerous other anthocyanins in low amount. Thanks to mass spectrometry data, eleven different compounds were characterized; their maximum absorption wavelength and their mass fragmentations are displayed in Table 1. A tentative identification of the core anthocyanin of each compound was realized, and shows a majority of pelargonidin-derived pigments in RSWP.

[0081] Mass spectrometry revealed the presence of two different anthocyanins under the major peak, this peak being principally made up of the compound based on pelargonidin (m/z 877).

[0082] This first characterization of anthocyanin from RSWP showed the presence of two major anthocyanins referred to herein as Anthocyanin 1 and Anthocyanin 2, respectively. These compounds have molecular weight of 877 and 1039 g/mol and are both based on pelargonidin. Their characteristics are shown in Table 1, together with other pelargonidin-based anthocyanins found in minor amounts in RSWP.

TABLE-US-00002 TABLE 1 Rt .sub.MAX m/z Fragment ions Associated (min) (nm) [M + H].sup.+ (m/z) pelargonidin 28.8 504 877 715, 433, 271 Anthocyanin 1 33.4 504 933 771, 433, 271 other 35.3 526 919 757, 433, 271 other 37.6 508 1039 877, 433, 271 Anthocyanin 2 39.6 508 1095 933, 271 other

[0083] Based on the areas of peaks visible on FIG. 1 the content of pelargonidin-based anthocyanins present in the extract was found to be about 58 mol-%. The amount of Anthocyanin 1 and Anthocyanin 2, based on all anthocyanins, was estimated to be 46.1 mol-%.

Example 2

Extraction and Isolation of the Major Anthocyanins

[0084] A RSWP concentrated extract at 8 CU/kg was extracted using ethyl acetate in order to remove phenolic compounds except anthocyanins. A volume of 30 ml of concentrated extract was diluted in 270 ml of acidified water (pH3) and then washed three times with 300 ml of ethyl acetate. The aqueous phase was concentrated under vacuo.

[0085] The anthocyanin extract obtained was purified on a Sephadex LH20 column (400 mm260 mm, Pharmacia), being eluted with acetic acid/water (4.6:100, v/v) at a flow rate of 16 ml/h. Two red-colored bands were collected. All fractions collected were analyzed by HPLC and fractions of high purity 85%) were grouped together.

[0086] Analytical HPLC was performed using a LiChrosorb RP-18 Column (250 mm4.6 mm, 5.0 m) by injecting 10 l of the filtered extracts. A combination of two solvents was used for elution: A, Water/HCOOH/Acetonitrile (87/10/3, v/v/v) and B, Water/HCOOH/Acetonitrile (40/10/50, v/v/v). The column flow was set at 0.8 ml/min. and the column temperature at 30 C. The gradient used was as summarized below:

TABLE-US-00003 Time Solvent A Solvent B (min) (vol.-%) (vol.-%) 0 94 6 20 80 20 35 60 40 40 40 60 45 10 90 50 10 90

[0087] Fractions containing high purity of one of the two anthocyanins of interest were grouped together and then concentrated under vacuo before freeze-drying. Powders of each anthocyanin were analyzed by NMR.

Example 3

NMR Analyses

A) Methods

[0088] The NMR experiments (.sup.1H, COSY, ROESY, HSQC, HSQC-TOCSY, HMBC, .sup.13C) were obtained at 600.13 MHz on a BRUKER Avance II 600 instrument equipped with a TCI .sup.1H.sup.13C/1.sup.5N CryoProbe at 27 C. Dried samples were solubilized in 500 l in DMSO-d6-TFA-d 99.99% 90:10.

B) Results and Discussion

[0089] Anthocyanins 1 and 2 were isolated in sufficient amounts to be characterized by NMR. Their structures were identified by .sup.1H and .sup.13C NMR spectroscopy in DMSO/TFA (90:10). Tables 2 and 3 show the .sup.1H and .sup.13C assignment and HSQC and TOCSY correlations of Anthocyanin 1 and Anthocyanin 2, respectively. Therein un=unresolved, s=singlet, d=doublet and t=triplet.

TABLE-US-00004 TABLE 2 Anthocyanin 1 ATOM .sub.13C.sup.a .sub.1H.sup.b Correlations Pelargonidin 1 2 162.7 3 144.3 4 135.3 8.94; s.sup. ROESY (Glc-a-1) 5 155.4 6 104.3 6.97; d (1.5) ROESY (Glc-c-1) 7 168.1 8 96.4 7.09; d (1.5) 9 155.7 10 111.9 .sup.1 119.3 .sup.2 135.3 8.58; d (8.8) .sup.3 117.1 7.07; d (8.8) .sup.4 165.4 .sup.5 117.1 7.07; d (8.8) .sup.6 135.3 8.58; d (8.8) Glucose-a HSQC-TOCSY (100.1; 80.3; 77.6; 76.5; 69.5; 60.7) 1 100.1 5.46; d (7.7) HMBC (Pelar-3), ROESY (Glc-a-2, 3, 5, Pelar-4) 2 80.3 3.92; t (8.4) HMBC (Glc-a-1, 3, Glc-b-1), ROESY (Glc-b-1) 3 76.5 3.63; t (9.0) 4 69.5 3.30; t (8.8) 5 77.6 3.49; un.sup. 6 60.7 3.69; d (10) 3.46; un.sup. Glucose-b HSQC-TOCSY (104.0; 76.5; 74.6; 74.3; 69.8; 63.2) 1 104.0 4.84; d (7.7) HMBC (Glc-3), ROESY (Glc-a-2, Glc-b-2, 3, 5) 2 74.6 3.08; t (8.8) 3 76.5 3.24; t (8.8) 4 69.8 3.27; un.sup. 5 74.3 3.23; un.sup. 6 63.2 4.14; dd (12.5; 2.2) 4.09; dd (11.7; 4.8) Glucose-c HSQC-TOCSY (101.4; 77.6; 76.0; 73.2; 69.7; 60.7) 1 101.4 5.12; d (7.7) HMBC (Pelar-5), ROESY (Glc-c-2, 3, 5, Pelar-6) 2 77.6 3.46; un.sup. 3 76.0 3.36; t (9.2) 4 69.7 3.27; un.sup. 5 77.6 3.46; un.sup. 6 60.7 3.74; dd (13.3; 2.2) 3.54; dd (11.9; 5.3) p-hydroxybenzoate 1 165.5 2 120.3 3 131.5 7.56; d (8.8) 4 115.3 6.67; d (8.8) 5 161.9 6 115.3 6.67; d (8.8) 7 131.5 7.56; d (8.8) Pelargonidin 1 2 162.7 3 144.3 4 135.3 8.94; s.sup. ROESY (Glc-a-1) 5 155.4 6 104.3 6.97; d (1.5) ROESY (Glc-c-1) 7 168.1 8 96.4 7.09; d (1.5) 9 155.7 10 111.9 .sup.1 119.3 .sup.2 135.3 8.58; d (8.8) .sup.3 117.1 7.07; d (8.8) .sup.4 165.4 .sup.5 117.1 7.07; d (8.8) .sup.6 135.3 8.58; d (8.8) Glucose-a HSQC-TOCSY (100.1; 80.3; 77.6; 76.5; 69.5; 60.7) 1 100.1 5.46; d (7.7) HMBC (Pelar-3), ROESY (Glc-a-2, 3, 5, Pelar-4) 2 80.3 3.92; t (8.4) HMBC (Glc-a-1, 3, Glc-b-1), ROESY (Glc-b-1) 3 76.5 3.63; t (9.0) 4 69.5 3.30; t (8.8) 5 77.6 3.49; un.sup. 6 60.7 3.69; d (10) 3.46; un.sup. Glucose-b HSQC-TOCSY (104.0; 76.5; 74.6; 74.3; 69.8; 63.2) 1 104.0 4.84; d (7.7) HMBC (Glc-3), ROESY (Glc-a-2, Glc-b-2, 3, 5) 2 74.6 3.08; t (8.8) 3 76.5 3.24; t (8.8) 4 69.8 3.27; un.sup. 5 74.3 3.23; un.sup. 6 63.2 4.14; dd (12.5; 2.2) 4.09; dd (11.7; 4.8) Glucose-c HSQC-TOCSY (101.4; 77.6; 76.0; 73.2; 69.7; 60.7) 1 101.4 5.12; d (7.7) HMBC (Pelar-5), ROESY (Glc-c-2, 3, 5, Pelar-6) 2 77.6 3.46; un.sup. 3 76.0 3.36; t (9.2) 4 69.7 3.27; un.sup. 5 77.6 3.46; un.sup. 6 60.7 3.74; dd (13.3; 2.2) 3.54; dd (11.9; 5.3) p-hydroxybenzoate 1 165.5 2 120.3 3 131.5 7.56; d (8.8) 4 115.3 6.67; d (8.8) 5 161.9 6 115.3 6.67; d (8.8) 7 131.5 7.56; d (8.8)

TABLE-US-00005 TABLE 3 Anthocyanin 2 ATOM .sub.13C.sup.a .sub.1H.sup.b Correlations Pelargonidin 1 2 163.0 3 144.2 4 135.2 8.83; s.sup. ROESY (Glc-a-1) 5 155.4 6 104.9 6.93; d (1.8) ROESY (Glc-c-1) 7 168.3 8 96.1 9.97; s.sup. 9 155.6 10 112.0 .sup.1 119.3 .sup.2 135.2 8.50; d (8.8) .sup.3 117.2 7.04; d (8.8) .sup.4 165.5 .sup.5 117.2 7.04; d (8.8) .sup.6 135.2 8.50; d (8.8) Glucose-a HSQC-TOCSY (100.3; 81.3; 76.1; 74.3; 69.9; 63.3) 1 100.3 5.54; d (7.3) HMBC (Pelar-3), ROESY (Glc-a-2, 3, 5, Pelar-4) 2 81.3 3.95; t (8.1) HMBC (Glc-a-2), ROESY (Glc-a-2, Glc-b-2, 3, 5) 3 76.1 3.69; t (8.8) 4 69.9 3.42; t (9) 5 74.3 3.83 6 63.3 4.37; d (12.5) HMBC (trans-caffeoyl-1), ROESY 2.25; dd (11.8; 6.2) (Glc-a-4, 5) Glucose-b HSQC-TOCSY (104.4; 76.4; 74.8; 74.3; 69.9; 63.1) 1 104.4 4.78; d (7.7) HMBC (Glc-a-2), ROESY (Glc-a-2, Glc-b-2, 3, 5) 2 74.8 3.14; t (8.4) 3 76.4 3.25; un.sup. 4 69.9 3.25; un.sup. 5 74.3 3.25; un.sup. 6 63.1 4.11; d (11) 4.06; dd (11.7; 4.0) Glucose-c HSQC-TOCSY (102.0; 77.9; 76.1; 73.4; 69.9; 61.0) 1 102.0 5.09; d (8.1) HMBC (Pelar-5), ROESY (Glc-c-2, 3, 5, Pelar-6) 2 73.4 3.51; t (8.4) 3 76.1 3.36; t (9.9) 4 69.9 3.25; un.sup. 5 77.9 3.48 6 61.0 3.77; d (11.0) 3.54; dd (12.0; 5.9) p-hydroxybenzoat 1 165.5 2 120.5 3 131.5 7.49; d (8.8) 4 115.3 6.62; d (8.8) 5 162.1 6 115.3 6.62; d (8.8) 7 131.5 7.49; d (8.8) trans-caffeoyl 1 166.8 2 113.8 6.09; d (15.9) 3 145.8 7.27; d (15.9) 4 125.6 5 115.4 6.91; d (1.5) 6 145.6 7 148.6 8 116.0 6.73; d (8.1) 9 121.6 6.81; dd (8.4; 1.8)

[0090] From the above data Anthocyanin 1 was identified as pelargonidin 3-O-(2-O-(6-O-para-hydroxybenzoyl-glucopyranosyl)-glucopyranoside)-5-O-glucopyranoside:

##STR00006##

[0091] Similarly, Anthocyanin 2 has been identified as pelargonidin 3-O-(2-O-(6-O-para-hydroxybenzoyl-glucopyranosyl)-6-O-trans-caffeoyl-glucopyranoside)-5-O-glucopyranoside:

##STR00007##

[0092] These structures are in agreement with mass spectrometry data and UV-visible spectra discussed above.

Example 4

[0093] Shade and Stability of Liquid Bulk Made from RSWP

[0094] RSWP liquid concentrate is evaluated for its shade against other anthocyanin references and for stability during cold storage.

A) Preparation of RSWP Concentrate

[0095] Sliced tubers were exhausted through extractions with acidified water. After clarification step, filtrate was purified onto an absorbent resin. Final concentration leads to a product at 65 Brix.

B) Color Evaluation and Stability Test

[0096] Shade conferred by RSWP concentrate was evaluated against other anthocyanin reference, the red radish, having similar shade but presenting off-flavors (sulfur compounds).

[0097] Samples of liquid concentrate were stored in cold room at 4-8 C. during 6 months including regular analytical evaluation. Samples were sacrificed after each evaluation. Bulk stability was evaluated through spectrophotometric and colorimetric measurements, turbidity, and amount of sludges.

[0098] Spectrophotometric measurements were performed in 1 cm-length quartz cell in a pH3 buffer using spectrophotometer HP8354. Red sweet potato concentrate was characterized through color strength E.sub.3 (expressed in color unit/kg).

[0099] Colorimetric measurements were performed in 1 cm-length quartz cell in a pH3 buffer using Spectraflash 650 (Datacolor) in transmission mode under D65 illuminant 10 Deg. Turbidity was measured on a VWR turbidimeter.

C) Results

[0100] Table 4 provides comparative color parameters of RSWP concentrate and red radish powder, and Table 5 shows the cold storage stability of RSWP concentrate.

TABLE-US-00006 TABLE 4 L* C h RSWP concentrate 70.0 62 23 Red Radish powder 70.0 63 17

[0101] RSWP concentrate presents similar brightness as pure red radish but its shade is more red-orange.

TABLE-US-00007 TABLE 5 E.sub.3 (CU/kg) L* C h Turbidity Sludges t0 8.2 0.1 70 62 23 <1 NTU <0.1% t0 + 1 mth 8.6 0.1 70 61 23 <1 NTU <0.1% t0 + 2 mths 8.0 0.1 70 62 24 <1 NTU <0.1% t0 + 4 mths 8.3 0.1 70 62 23 <1 NTU <0.1% t0 + 5 mths 8.3 0.1 70 61 23 <1 NTU <0.1% t0 + 6 mths 8.1 0.1 70 62 23 <1 NTU <0.1%

[0102] RSWP concentrate kept in cold conditions is highly stable considering color as well as physico-chemical parameters.

Example 5

[0103] Stability in Beverage Application of a Color Made from RSWP

[0104] RSWP concentrate is evaluated in a model beverage medium submitted to a pasteurization step for determining cold, heat and light stabilities against two standard references having similar shades and being used in this application.

A) Preparation of Colored Model Beverage Medium

[0105] The model beverage medium was prepared according to the following recipe.

TABLE-US-00008 Saccharose 43.00% Potassium Sorbate 0.09% Sodium Benzoate 0.07% Citric acid anhydrous 0.86% Milli Q water 55.98%

[0106] A soft drink concentrate around 40 Brix was obtained and further diluted with Milli Q water until 11 Brix. The pH was finally adjusted to 3.00.2 with citric acid.

[0107] As colorant RSWP concentrate at 0.22% was added directly into the model beverage medium. For comparison a red radish/black carrot anthocyanin blend (referred to as rr/bc hereinafter) at 0.13% and having basically the same color shade was used. As a reference (DE* 2000=0) 8.2 wt.-% carminic acid was used in an amount of 0.4 wt.-% After submission to a pasteurization step (referred to as HT) at 92 C. for 40 seconds, the colored beverages were poured into PET bottles and then stored under the following conditions:

For light stability: daylight exposure, room temperature
For heat stability: in a binder incubator at 40 C., 65% RH
For reference storage: in a cold room at 4 C. in the dark

[0108] Colorimetric follow-up was done every week during one month and then after 2-month storage. Measurements were performed directly on the PET bottles using Spectraflash 650 (Datacolor) in transmission mode under D65 illuminant 10 Deg.

B) Results

[0109] Table 6 summarizes the shades of the model beverage medium colored with RSWP or the references at day 0.

TABLE-US-00009 TABLE 6 L* C h DE* 2000 Carminic acid 0.4% 38.80 94.02 44.64 RSWP concentrate 39.26 92.89 45.90 1.15 red radish/black carrot (rr/bc) 39.46 90.88 45.14 0.93 *DE* 2000 is an indicator for the total color variation, which includes the changes all of L*, C and h values and illustrates the total color difference. High values indicate large differences.

[0110] Beverage colored with RSWP is slightly duller than the one colored with carminic acid but is brighter than the beverage colored with rr/bc. Shades brought by the RSWP concentrate and the comparison (rr/bc) and the reference (carminic acid) are similar.

[0111] Table 7 shows the color stability after pasteurization (HT) of the model beverage medium colored with RSWP or rr/bc.

TABLE-US-00010 TABLE 7 L* C h DE* 2000 Carminic acid before HT 38.80 94.02 44.64 Carminic acid after HT 38.62 93.59 44.65 0.18 RSWP before HT 39.26 92.89 45.90 RSWP after HT 39.55 93.35 46.00 0.28 red radish/black carrot before HT 39.46 90.88 45.14 red radish/black carrot after HT 40.00 90.96 45.18 0.47

[0112] Beverage colored with RSWP is more stable through pasteurization compared to the beverage colored with rr/bc. However it remains slightly more sensitive than a beverage colored with carminic acid.

[0113] FIG. 5 shows the evolution of DE* 2000 along the 2-month storage of colored model beverage medium in cold room.

[0114] Beverages colored with carminic acid, RSWP or rr/bc, respectively, present similar stabilities under cold storage. Evolution of coloration is not visually detected whatever the color reference.

[0115] The evolution of DE* 2000 during the 2-month storage of colored model beverage medium under (i) light exposure and (ii) heat exposure is shown in FIGS. 6 and 7, respectively.

[0116] The beverage colored with RSWP is as stable as the one colored with carminic acid after 2-month light exposure (no visual shift of shade is detected in both cases) and is far more stable in both tests than a beverage colored with rr/bc, which undergoes a wide evolution of coloration.

Example 6

[0117] Impact of Ascorbic Acid on the Color Stability of Beverage colored with RSWP extract

A) Experimental

[0118] RSWP concentrate is evaluated in a model beverage medium containing ascorbic acid for determining cold, heat and light stabilities against a standard reference having similar shade.

[0119] The model beverage medium prepared in Example 5 was used, except that 250 ppm ascorbic acid was added before final adjustment of pH to 3.00.2 with citric acid.

[0120] The colors were added in the same manner as described in Example 5, and then the colored beverages were poured into PET bottles and stored under the conditions defined in Example 5. Colorimetric follow-up and measurements were then also performed as defined in Example 5.

B) Results

[0121] FIG. 8 shows the evolution of DE* 2000 along the 2-month storage of colored model beverage medium in cold room.

[0122] Beverages colored with carminic acid, RSWP or rr/bc, respectively, present similar stabilities under cold storage. Evolution of coloration is not visually detected whatever the color reference.

[0123] The evolution of DE* 2000 during 2-month storage of colored model beverage medium under (i) light exposure and (ii) heat exposure is shown in FIGS. 9 and 10, respectively.

[0124] Beverage colored with RSWP is as stable as the one colored with carminic acid after 2-month light exposure in presence of ascorbic acid and the shift of shade is limited. Contrary thereto the beverage colored with rr/bc present a far lower stability to light exposure in presence of ascorbic acid.

[0125] Also, the beverage colored with RSWP, while presenting a not yet optimal stability such as carminic acid, provides a large improvement over rr/bc, considering the evolution of shade associated with the use of this traditional comparative colorant.

Example 7

Color Stability of a RSWP-Colored Fruit Preparation

[0126] RSWP concentrate is evaluated in a fruit preparation application for determining stabilities during storage of the fruit preparation itself and storage of a blend fruit preparation/white mass against a standard reference being used in this application.

[0127] Compared stabilities to pasteurization step of blends fruit preparation/white mass are also described.

A) Ingredients and Process

[0128] The model fruit preparation is a strawberry fruit preparation at pH 3.82 and 40.3 Brix. The colors were added directly into the fruit preparation at following dosages:

RSWP concentrate at 0.9%
Solubilized carmine lake at 0.56%

[0129] Model white mass is a commercial product containing 3.5% fat.

[0130] Colored fruit preparation was incorporated into the white mass at a weight ratio of 15/85 and the mixture was further pasteurized at 90 C. for 5 minutes.

B) Stability Evaluation

[0131] The colored fruit preparations were stored during one month at 10 C. and mixtures of fruit preparation/white mass were stored for 14 days in a cold room at 4 C. in the dark.

[0132] Colorimetric follow-up was done every week during two weeks for blends fruit preparation/white mass, and after one month storage for fruit preparations alone. Measurements were performed in Petri boxes using Datacolor SF 450 in reflection mode.

C) Results

[0133] Table 8 summarizes the shades of the blends fruit preparation/white mass colored with RSWP or the carmine lake reference at day 0 before the pasteurization step.

TABLE-US-00011 TABLE 8 L* C h DE* 2000 RSWP 75.52 15.37 10.94 carmine lake 73.61 21.75 4.93 4.52

[0134] The blend colored with RSWP is duller and more orange that the one colored with carmine lake.

[0135] Table 9 shows the stability of blends fruit preparation/white mass colored with RSWP or the carmine reference during the pasteurization step.

TABLE-US-00012 TABLE 9 L* C h DE* 2000 RSWP before HT 75.52 15.37 10.94 RSWP after HT 76.27 13.68 15.56 1.69 Carmine lake before HT 73.61 21.75 4.93 Carmine lake after HT 75.86 16.50 15.66 4.51

[0136] Blend fruit preparation/white mass colored with RSWP concentrate is far more stable through pasteurization compared to the blend colored with carmine lake and the variation of shade is acceptable (DE* 2000 below 2). This implies that the difference of shade between the two fruit preparation/white mass blends is reduced after pasteurization (DE* 2000=2.04).

[0137] FIG. 11 shows the evolution of DE* 2000 during the 2-week storage of fruit preparation/white mass blends in cold room at 4 C.

[0138] Blend fruit preparation/white mass colored with RSWP concentrate is less stable compared to the one colored with carmine lake under cold storage. Evolution of shade is visually detectable in former case but is considered as acceptable based on the DE 2000 value below 2.

[0139] FIG. 12 shows the evolution of DE* 2000 during the 1-month storage of the colored fruit preparation.

[0140] Fruit preparation colored with RSWP concentrate is almost as stable as the one colored with carminic acid after 1-month cold storage at 10 C. and no visual shift of shade is detected in both cases.