U.S. patent application number 17/423624 was filed with the patent office on 2022-03-24 for a colored beverage having a neutral ph.
The applicant listed for this patent is GNT GROUP B.V.. Invention is credited to Elena LEEB, Jane Lee MACDONALD.
Application Number | 20220087291 17/423624 |
Document ID | / |
Family ID | 1000006061471 |
Filed Date | 2022-03-24 |
United States Patent
Application |
20220087291 |
Kind Code |
A1 |
MACDONALD; Jane Lee ; et
al. |
March 24, 2022 |
A COLORED BEVERAGE HAVING A NEUTRAL PH
Abstract
A liquid foodstuff including a. a spirulina-extract having
phycocyanin, b. at least one multi-sulphated carrageenan and/or
pectin c. a chelating agent d. a solvent, wherein the amount of
multi-sulphated carrageenan and/or pectin is between 0.003 and 0.6
wt %, wherein the weight ratio between multi-sulphated carrageenan
and/or pectin and phycocyanin ranges between 1:1 and 100:1, wherein
the chelating agent is present between 1-2000 ppm, wherein the
solvent is water and optionally ethanol, wherein the amount of
solvent ranges between 60-99 wt %, wherein the wt % are relative to
the total weight of the liquid foodstuff.
Inventors: |
MACDONALD; Jane Lee;
(Mierlo, NL) ; LEEB; Elena; (Mierlo, NL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
GNT GROUP B.V. |
MIERLO |
|
NL |
|
|
Family ID: |
1000006061471 |
Appl. No.: |
17/423624 |
Filed: |
November 19, 2019 |
PCT Filed: |
November 19, 2019 |
PCT NO: |
PCT/EP2019/081840 |
371 Date: |
July 16, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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62794083 |
Jan 18, 2019 |
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62794094 |
Jan 18, 2019 |
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62794104 |
Jan 18, 2019 |
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62794074 |
Jan 18, 2019 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23L 29/256 20160801;
A23L 2/44 20130101; A23L 2/46 20130101; A23L 2/58 20130101; A23L
29/231 20160801; A23L 5/46 20160801; A23V 2002/00 20130101; C12G
3/04 20130101 |
International
Class: |
A23L 2/58 20060101
A23L002/58; A23L 5/46 20060101 A23L005/46; A23L 2/46 20060101
A23L002/46; A23L 2/44 20060101 A23L002/44; A23L 29/256 20060101
A23L029/256; A23L 29/231 20060101 A23L029/231; C12G 3/04 20060101
C12G003/04 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 11, 2019 |
EP |
19156394.9 |
Feb 11, 2019 |
EP |
19156427.7 |
Feb 11, 2019 |
EP |
19156440.0 |
Feb 11, 2019 |
EP |
19156444.2 |
Claims
1. A liquid foodstuff, comprising: a. a spirulina-extract
comprising phycocyanin b. at least one multi-sulphated carrageenan
and/or pectin c. a chelating agent d. a solvent, i. wherein the
amount of the at least one multi-sulphated carrageenan and/or the
pectin is between 0.003 and 0.6 wt %, ii. wherein the weight ratio
between the at least one multi-sulphated carrageenan and/or the
pectin and the phycocyanin ranges between 1:1 and 100:1, iii.
wherein the chelating agent is present between 1-2000 ppm, iv.
wherein the solvent consists of water and optionally ethanol, v.
wherein the amount of the solvent ranges between 60-99 wt %, vi.
wherein the wt % are relative to the total weight of the liquid
foodstuff.
2. The liquid foodstuff according to claim 1, wherein the solvent
content is between 75-98 wt %.
3. The liquid foodstuff according to claim 1, wherein the solvent
consists of water and optionally ethanol (ethylalcohol).
4. The liquid foodstuff according to claim 1, wherein the foodstuff
contains between 0.1-20 wt % ethanol.
5. The liquid foodstuff according to claim 1, wherein the at least
one multi-sulphated carrageenan is a non-degraded occurring
carrageenan.
6. The liquid foodstuff according to claim 1, wherein the at least
one multi-sulphated carrageenan is a hydrolyzed carrageenan.
7. The liquid foodstuff according to claim 1, wherein the at least
one multi-sulphated carrageenan is selected from the group
consisting of -carrageenan, .delta.-carrageenan, .mu.-carrageenan,
.theta.-carrageenan, .lamda.-carrageenan and .nu.-carrageenan, a
hydrolysed product of -carrageenan, a hydrolysed product of
.delta.-carrageenan, a hydrolysed product of .mu.-carrageenan, a
hydrolysed product of .theta.-carrageenan, a hydrolysed product of
.lamda.-carrageenan and a hydrolysed product of
.nu.-carrageenan.
8. The liquid foodstuff according to claim 1, wherein the at least
one multi-sulphated carrageenan content of the liquid foodstuff is
between 0.003-0.6 wt %.
9. The liquid foodstuff according to claim 1, wherein the pectin
content of the liquid foodstuff is between 0.003-0.6 wt %.
10. The liquid foodstuff according to claim 1, wherein the
phycocyanin content is between 0.003-0.07 wt %.
11. The liquid foodstuff according to claim 1, wherein the
dissolved at least one multi-sulphated carrageenan and/or the
pectin and the phycocyanin present within the liquid foodstuff at a
weight ratio between 2:1-50:1.
12. The liquid foodstuff according to claim 1, wherein the
chelating agent is selected from the group consisting of ethylene
diamine tetra acetic acid and/or its Na, K, Ca salts and L-glutamic
acid N,N-diacetic acid tetrasodium salt (GLDA).
13. The liquid foodstuff according to claim 1, wherein the
chelating agent is ethylene diamine tetra acetic acid, which is
present between 1-100 ppm.
14. The liquid foodstuff according to claim 13, wherein ethylene
diamine tetra acetic acid and the phycocyanin present within the
liquid foodstuff at a weight ratio between 2:1-1:300.
15. Process for preparing the liquid foodstuff according to claim
1, wherein the process comprises the steps: a. adding and
dissolving the at least one multi-sulphated carrageenan and/or the
pectin in the water and mixing until the at least one
multi-sulphated carrageenan and/or the pectin are dissolved, as
determined by visual observation; b. adding the phycocyanin, and
mixing until dissolved at a pH of at least 5; c. adding a chelating
agent; d. optionally adding a sweetener, flavor, vitamin, mineral,
salt, buffer, juice, or other beverage components; e. optionally
adding other pigments; f. adding the solvents to obtain the total
volume; g. treating the mixture of a)-f) by either thermally
processing the liquid to at least 65.degree. C. and filling it into
a container; filling the liquid into a container and thermally
processing the filled package to at least 100.degree. C. or no
thermal treatment and cold filling into a container and controlling
by formulation (alcohol or water activity) addition of
preservatives or high pressure.
16. The liquid foodstuff according to claim 1, wherein the at least
one multi-sulphated carrageenan is selected from the group
consisting of -carrageenan, .lamda.-carrageenan, a hydrolysed
product of -carrageenan, and a hydrolysed product of
.lamda.-carrageenan.
17. The liquid foodstuff according to claim 1, wherein the at least
one multi-sulphated carrageenan is selected from the group
consisting of .lamda.-carrageenan and hydrolyzed
.lamda.-carrageenan.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to a color stabilized liquid
foodstuff containing a phycocyanin from spirulina-extract.
BACKGROUND OF THE INVENTION
[0002] Beverages and colored beverages are known in the art. The
use of natural food pigments over synthetic ones has increased in
the food industry, due to increased consumer demands. While several
natural pigments have been applied to low acid beverages, having a
pH greater than 4.6, there is a need for a stable blue pigment
which can be used on its own or blended with other pigments to
deliver a range of colors including but not limited to green,
purple, brown, and black.
[0003] US2018271119 discloses the use of phycocyanin derived from
Galdieria sulphuraria, which is capable of growing at both high
temperature and low pH, for use in beverages. The increased acid
stability of phycocyanin (and allophycocyanin) extracted from
Galdieria sulphuaria might be explained by the differences in the
amino acid sequence in comparison to phycocyanin from commercially
available spirulina (such as Arthrospira platensis, Arthrospira
maxima, ect.). But Galdieria sulphuraria phycocyanin extracts have
not yet been approved or undergone long term testing, as Spirulina
derived phycocyanin has.
[0004] Selig, et al. discloses the use of beet-pectin to stabilize
phycocyanin from a Spirulina-extract at pH 6.8 in an aqueous
solution. Beet-pectin appears to be effective at pH 6.8, but is not
stable upon thermal treatment, up to 120.degree. C. which is common
in the food industry.
[0005] Dewi, et al. discloses a spirulina-extract encapsulated in
K-carrageenan for use in an aqueous solution.
[0006] Phycocyanin, from a Spirulina-extract, is to-date the only
natural blue pigment approved by the US-FDA (FR Doc No: 2013-19550)
and European Union as a coloring food. It is sold in liquid or in
powder form for use as blue pigment in foods. Phycocyanin, however,
has the disadvantage of being unstable in aqueous systems between
pH 2.7-6 where protein aggregation occurs, and also instable under
thermal treatment, leading to loss of color which limits the use of
phycocyanin in low acid foodstuffs. Thus, phycocyanin has limited
use for its food coloring properties in beverages, such as in for
example non-alcoholic and alcoholic drinks, which have a pH greater
than 4.6. Health conscious customers demand a wide range of
naturally colored beverages, and phycocyanin is to-date the only
available natural blue pigment. Hence, there is a need to stabilize
phycocyanin when used in beverages which undergo thermal treatment
or in cold filled alcoholic beverages.
SUMMARY OF THE INVENTION
[0007] The inventors have found a solution to stabilize the blue
color, from a phycocyanin containing spirulina-extract, in liquid
foodstuffs.
The present invention relates to a liquid foodstuff comprising
[0008] a. a spirulina-extract comprising phycocyanin, [0009] b. at
least one multi-sulphated carrageenan and/or pectin [0010] c. a
chelating agent [0011] d. a solvent, [0012] wherein the amount of
multi-sulphated carrageenan and/or pectin is between 0.003 and 0.6
wt %, [0013] wherein the weight ratio between multi-sulphated
carrageenan or pectin and phycocyanin ranges between 1:1 and 100:1,
[0014] wherein the chelating agent is present between 1-2000 ppm,
[0015] wherein the solvent consists of water and optionally
ethanol, [0016] wherein the amount of solvent ranges between 60-99
wt %, [0017] wherein the wt % are relative to the total weight of
the liquid foodstuff.
[0018] The liquid foodstuff according to the invention has an
increased color stability over the heating process as well as
storage duration. The negative effects are
aggregation/precipitation, color fading, color hue changes and lack
of stability over time (weeks or months), as foodstuffs need to
withstand long transport and varying storage conditions.
[0019] Advantages of the liquid foodstuff according to the
invention are color stability and prevention of
aggregation/precipitation; thereby enabling beverages to be
naturally colored; for example, in blue and blue-based colors.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1: Absorbance spectra of aqueous liquid foodstuff at pH
6.5 before and after UHT processing (135.degree. C. for 6 seconds)
with and without .lamda.-carrageenan and EDTA. The combination of
.lamda.-carrageenan with Na.sub.2 EDTA had the best color retention
post thermal processing.
DETAILED DESCRIPTION OF THE INVENTION
[0021] The liquid foodstuff of the invention contains
multi-sulphated-carrageenan and/or pectin, a phycocyanin containing
spirulina-extract and a chelator, preferably at a pH between 4.6
and 8.
[0022] The liquid foodstuff may further contain other components
like for example sweeteners, stabilizers, chelating agents, acids,
proteins, salts, flavors, vitamins, minerals, pigments, and
preservatives.
[0023] The liquid foodstuff may have a blue color due to the
phycocyanin content, but other colors based on blue may be obtained
by way of mixing other pigments in with the liquid foodstuff. The
liquid foodstuff may contain other pigments, such as safflomin
(safflower), anthocyanin, carotenoid, betanin, annatto, lycopene,
curcumin and chlorophyll. The pigments may be added to the blue
colored phycocyanin containing liquid foodstuff to blend into other
colors, such as for example green using safflomin.
[0024] The liquid foodstuff may contain additives and/or
preservatives, such as salts, sorbic acid, salt of sorbate, benzoic
acid, salt of benzoate, sodium hexametaphosphate, natamycin, nisin
and sulfites.
[0025] The liquid foodstuff preferably has a solvent content of
60-99 wt %, more preferably between 75-98 wt %, most preferably
between 83-97 wt %. The solvent consists of water and optionally
ethanol (ethylalcohol). When ethanol is present typically, the
liquid foodstuff contains between 0.1-20 wt %, preferably between
0.5-17 wt %, more preferably between 1-15 wt %, most preferably
between 3-10 wt % ethanol.
[0026] The wt % is defined as the weight % of a component relative
to the total weight of the liquid foodstuff, unless defined
otherwise.
[0027] Preferably the liquid foodstuff is a beverage, such as a
coconut water, alkaline water, coffee beverages, fruit or vegetable
drink, smoothie, dairy beverage, dairy alternatives, nut milks,
nutritional drink, and/or alcoholic beverage.
[0028] The liquid foodstuff's color can be required to be stable
over a long period of time (through transport and storage over
weeks or months), at pH greater than 4.6. The color also needs to
be stable after the liquid food product is subject to high
temperature, up to 120.degree. C., as these temperatures are
commonly used in the food industry in processes such as high
temperature short time pasteurization to ensure food safety.
[0029] Phycocyanin
[0030] The liquid foodstuff contains a phycocyanin containing
spirulina-extract.
[0031] The phycocyanin containing spirulina-extract can be a water
extract of for example Arthrospira platensis and Arthrospira
maxima. Spirulina is a cyanobacteria that contains carbohydrates,
lipids, fiber, minerals and amongst other proteins,
phycobiliproteins. The phycobiliproteins comprise of C-phycocyanin
and allophycocyanin. Phycocyanins exhibit the blue color of the
liquid foodstuffs according to the present invention. The total
phycocyanin content (which is the sum of the C-phycocyanin and
allophycocyanin) in phycocyanin containing spirulina-extracts may
vary by manufacturers and may typically range from 0.7-45.0 wt %.
It is also possible to apply higher concentrations of phycocyanins.
The phycocyanin containing spirulina-extract may also be known as
Spirulina color concentrate, as for example EXBERRY.RTM. Shade Blue
Powder. The phycocyanin containing spirulina-extract may be in
liquid or dry (powder or granule) form and may contain diluents
such as water, invert sugar, sucrose, and/or maltodextrin as
example. For use in this invention, liquid and dry forms of
phycocyanin containing spirulina-extract are applicable.
[0032] The liquid foodstuff preferably contains a dissolved
phycocyanin from a spirulina-extract.
[0033] Multi-Sulphated Carrageenan
[0034] The liquid foodstuff contains a multi-sulphated carrageenan
or combination of multi-sulphated carrageenan.
[0035] The liquid foodstuff preferably contains dissolved
multi-sulphated carrageenan. Carrageenans are viewed as dissolved,
when an aqueous solution is clear to the eye, and does not show
sediment or floating particulates.
[0036] Carrageenans are sourced from seaweed and they are highly
flexible molecules that can form curling helical structures.
Carrageenans are characterized as linear polysaccharides with
repeating galactose units. Carrageenans are classified by the
degree of sulfonation. Examples of mono-sulphated carrageenans are
.kappa.-carrageenan, .gamma.-carrageenan and .alpha.-carrageenan.
Examples of di-sulphated carrageenans are -carrageenan,
.delta.-carrageenan, .mu.-carrageenan and .theta.-carrageenan.
Examples of tri-sulphated carrageenans are .lamda.-carrageenan and
.nu.-carrageenan.
[0037] Multi-sulphated carrageenans are carrageenans containing at
least 2 sulphate groups, preferably 2 or 3 sulphate groups per
disaccharide unit.
[0038] In some embodiments the multi-sulphated carrageenan can be a
non-degraded carrageenan
[0039] In some embodiments the multi-sulphated carrageenan can be a
hydrolyzed multi-sulphated carrageenan preferably by thermal
treatment of a non-degraded multi-sulphated carrageenan under
acidic pH conditions.
[0040] Thermal treatment of carrageenan solutions at acidic pH
leads to a hydrolysis of the glycosidic bonds (.alpha.(1-3) and
.beta.(1-4) bonds) connecting the alternating D-galactose residues.
Thus, the average molecular weight was shown to decrease with
increasing hydrolysis duration (Singh & Jacobsson, 1994). In
addition, a good correlation of the measured viscosity and the
molecular weight was observed, meaning that with decreasing
molecular weight the viscosity decreases (Singh & Jacobsson,
1994). Preferably the pH is between 1.4 and 4 for hydrolysis of the
glycosidic bonds, and preferably the temperature is between
20-98.degree. C. for hydrolysis of the glycosidic bonds, more
preferably between 55 and 98.degree. C.
[0041] Due to the uniform backbone structure, the concentration of
carrageenan can be quantified by the content of galactose. In
addition, the quantification of galactose allows to determine if
the glyosidic bond or also the galactose monomer unit gets degraded
by the hydrolysis conditions.
[0042] A possibility to quantify galactose is the quantification of
monosaccharides after acid hydrolysis. Hydrolysis of carrageenan is
performed with 72% sulphuric acid (H.sub.2SO.sub.4) for 30 min at
30.degree. C., followed by further hydrolysis after dilution to 6%
sulphuric acid (H.sub.2OS.sub.4) for 3 hours at 100.degree. C., The
amount of monosaccharides rhamnose, galactosamine, arabinose,
glucosamine, galactose, glucose, mannose, xylose, galacturonic acid
and glucuronic acid in the hydrolysate can be quantitatively
analyzed by High Performance Anion Exchange Chromatography with
Pulsed Amperometric Detection (HPAEC-PAD) on a gold electrode.
Analyses are performed with an ICS-5000 DP pump, AS-AP autosampler,
DC column compartment and ED electrochemical detector (Thermo
Scientific). The neutral monosaccharides are eluted using a
gradient of 18 mM sodium hydroxide and 200 mM sodium hydroxide with
75 mM sodium acetate. Data analysis is done with Chromeleon
software version 7.2 (Thermo Scientific). Quantitative analyses are
carried out using standard solutions of the monosaccharides
(Sigma-Aldrich).
[0043] Analysis of galactose content of the used lambda carrageenan
powders shows a galactose content of 50-55 wt %.
[0044] Quantification of the galactose content in hydrolyzed
carrageenan solutions showed comparable galactose contents as
related to the initial amounts of used carrageenan powder. These
results demonstrate that a hydrolysis of carrageenan at
pH.gtoreq.1.5 and temperatures.gtoreq.95.degree. C. result in the
hydrolysis of the glycosidic bonds but do not degrade the galactose
unit itself.
[0045] In addition, the degree of sulphonation can be quantified by
the amount of Sulphur. A possibility to quantify Sulphur is based
on the combustion of the sample at temperatures above 1000.degree.
C. and reduction of the released Sulphur to Sulphur dioxide
(Elementar Vario Max Cube). The amount of Sulphur is quantified by
an IR detector which is calibrated using a calibration curve of
sulfadiazine.
[0046] The viscosity of the carrageenan solutions was characterized
by viscometry. Viscosity measurements were taken using an Anton
Paar Rheometer MCR 302 (Switzerland). Flow curves were measured
with a shear rate (1/s) range from 1 to 300 at 20.degree. C. To
compare different carrageenan solutions, the viscosity at a shear
rate of 100/s is used.
[0047] The preferred multi-sulphated carrageenans of the invention
are -carrageenan, .delta.-carrageenan, .mu.-carrageenan,
.theta.-carrageenan, .lamda.-carrageenan and .nu.-carrageenan, the
hydrolysed products of -carrageenan, .delta.-carrageenan,
.mu.-carrageenan, .theta.-carrageenan, .lamda.-carrageenan and
.nu.-carrageenan; more preferred are -carrageenan and
.lamda.-carrageenan, and its hydrolyzed products and most preferred
is .lamda.-carrageenan and hydrolyzed .lamda.-carrageenan.
[0048] Mixtures of multi-sulphated carrageenans can also be
used.
[0049] The multi-sulphated carrageenans to be used in the present
invention have preferably a viscosity between 1 and 3000 mPas, when
dissolved as a 4 wt % solution in water at a pH of 5.5.
[0050] The multi-sulphated carrageenan content of the liquid
foodstuff is between 0.003-0.6 wt %, preferably between 0.01-0.5 wt
%, more preferably between 0.02-0.3 wt %, most preferably 0.03-0.2
wt %.
[0051] The dissolved multi-sulphated carrageenans and phycocyanin
from a spirulina-extract are preferably present within the liquid
foodstuff at a weight ratio between 1:1-100:1, preferably between
2:1-50:1, more preferably between 3:1-25:1, most preferably between
4:1-15:1.
[0052] Pectin
[0053] Instead of using a multi-sulphated carrageenan, it is also
possible to apply pectin to stabilize the phycocyanin in
combination with a chelator. Pectin is a heteropolysaccharide
contained in the primary cell walls of plants. It is generally used
as a stabilizer, gelling agent, or thickener in foodstuff. Pectins
are classified as high-methoxy (HM) or low-methoxy (LM). The degree
of methylation determines the classification. High methoxy pectins
contain more than 50% of the carboxyl groups are methylated and
less than 50% methylation are called low methoxy (LM) pectins.
HM-pectins can form a gel under acidic conditions in the presence
of high sugar concentrations, while LM-pectins form gels by
interaction with divalent cations, particularly Ca.sup.2+. The
chemical composition of pectin varies based upon source material
with most common materials being citrus peels, apple pomace, and
sugar beet pulp. Regardless of source materials and degree of
methylation pectins are applicable to stabilize phycocyanin at
neutral pH and processing temperature greater than 80.degree. C.
when used in combination with a chelator.
[0054] The use of pectin in combination with a chelator shows an
unexpected synergistic effect of color stability upon heat
treatment.
[0055] The pectin content of the liquid foodstuff is between
0.003-0.6 wt %, preferably between 0.01-0.5 wt %, more preferably
between 0.02-0.3 wt %, most preferably 0.03-0.2 wt %.
[0056] The dissolved multi-sulphated carrageenans and/or pectin and
phycocyanin from a spirulina-extract are preferably present within
the liquid foodstuff at a weight ratio between 1:1-100:1,
preferably between 2:1-50:1, more preferably between 3:1-25:1, most
preferably between 4:1-15:1.
[0057] Color
[0058] The color performance is assessed using a spectrophotometer
and measuring absorbance at 620, 650 and 750 nm. The standard
method for phycocyanin determination was established by Yoshikawa
& Belay (2008) to calculate the native phycocyanin content
(mg/mL) from photometric measurements. It relies upon absorbance
measurements at 620 and 650 nm and the extinction coefficients of
C-phycocyanin and allophycocyanin at these wavelengths at pH
6.0.
[0059] The phycocyanin content in a spirulina-extract is calculated
using Yoshikawa & Belay (2008) method at pH 6.0 which is listed
below.
allophycocyanin .times. .times. ( mg .times. mL .times. ) = 0.180
.times. ( Abs .times. .times. 650 .times. .times. nm - Abs .times.
.times. 750 .times. .times. nm ) - 0.042 .times. ( Abs .times.
.times. 620 .times. .times. nm - Abs .times. .times. 750 .times.
.times. nm ) ##EQU00001## C - phycocyanin .times. .times. ( mg mL )
= 0.1 .times. 6 .times. 2 .times. ( Abs .times. .times. 620 .times.
.times. nm - Abs .times. .times. 750 .times. .times. nm ) - 0 . 0
.times. 9 .times. 8 .times. ( Abs .times. .times. 650 .times.
.times. nm - .times. Abs .times. .times. 750 .times. nm )
##EQU00001.2## Total .times. .times. Phycocyanin .times. .times. (
.times. mg mL ) = allophycocyanin .times. .times. ( .times. mg mL )
+ C - phycocyanin .times. .times. ( .times. mg mL )
##EQU00001.3##
[0060] To determine the amount of phycocyanin added to a liquid
foodstuff, the dosage level of phycocyanin containing
spirulina-extract is multiplied by the total phycocyanin content in
the phycocyanin containing spirulina-extract.
[0061] The phycocyanin content of the liquid foodstuff is
preferably between 0.003-0.07 wt %, preferably 0.006-0.05 wt %, and
most preferably 0.008-0.04 wt %.
[0062] The challenge with phycocyanins is the stability during or
after thermal treatment, which can lead to color loss and
precipitation. Precipitation occurs in the pH range of 2.7-6.0 and
for low acid foodstuff the critical range is between 4.6-6.0.
Precipitation can be avoided when the pH is above 6.0 or delayed
when the solution is in a gel or semi-solid form. The
aggregation/precipitation is assessed visually. High absorbance at
750 nm after blending or thermal processing indicates a strong
potential for protein aggregation.
[0063] Preferably the pH of the beverage according to the invention
is between 4.6-10, more preferably between 5-9, most preferably
between 5.5-8.
[0064] Chelators
[0065] The liquid foodstuff contains at least one chelator, which
may also be referred to as chelating agents.
[0066] Chelators are binding agents that influence the chemical
and/or physical state of the molecules/atoms they bind by forming
chelates. Chelators can improve color retention and have been found
to work synergistically with the multi-sulphated carrageenans and
pectin.
[0067] Chelating agents can be synthetic and natural compounds and
include the group of ethylene diamine tetra acetic acid and/or its
Na, K, Ca salts (EDTA), L-glutamic acid N,N-diacetic acid
tetrasodium salt (GLDA), galactaric acid, sodium hexametaphosphate,
glutathione, metallotheionein, 2,3-dimerapto-1-propanesulfonic
acid, chlorella, garlic, cilantro, selenium, milk thistle, vitamin
C, vitamin E, citrates, grape seed extract, quercetin, and lipoic
acid.
[0068] The chelating agents are preferably selected from the group
of ethylene diamine tetra acetic acid and/or its Na, K, Ca salts
(EDTA) and L-glutamic acid N,N-diacetic acid tetrasodium salt
(GLDA). EDTA is considered to be ethylene diamine tetra acetic acid
together with its Na, K, Ca salts, like for example calcium
disodium ethylenediaminetetraacetate, disodium
ethylenediaminetetraacetate, tetrasodium
ethylenediaminetetraacetate, dipotassium
ethylenediaminetetraacetate, and tripotassium
ethylenediaminetetraacetate.
[0069] Calcium disodium ethylenediaminetetraacetate is abbreviated
to CaNa.sub.2 EDTA, disodium ethylenediaminetetraacetate is
abbreviated to Na.sub.2 EDTA, tetrasodium
ethylenediaminetetraacetate is abbreviated to Na.sub.4 EDTA,
dipotassium ethylenediaminetetraacetate is abbreviated to K.sub.2
EDTA, and tripotassium ethylenediaminetetraacetate is abbreviated
to K.sub.3 EDTA.
[0070] The chelating agent is preferably present between 1-2000 ppm
in the liquid foodstuff.
[0071] 1 ppm of chelating agent is to be understood as 0.0001 wt %;
for example, 30 ppm of EDTA are 0.003 wt %.
[0072] EDTA is preferably present between 1-100 ppm, preferably
10-50 ppm, most preferably 20-40 ppm.
[0073] The EDTA and phycocyanin from a spirulina-extract are
preferably present within the liquid foodstuff at a weight ratio
between 2:1-1:300, preferably between 1:1-1:25, most preferably
1:2-1:15.
[0074] The multi-sulphated carrageenans and/or pectin in
combination with a chelator such as EDTA showed a surprisingly high
increase in color retention and extended shelf life after thermal
treatment and/or in the presence of ethyl alcohol at neutral
pH.
[0075] In an embodiment the liquid foodstuff is a beverage
comprising between 0.003-0.6 wt % of .lamda.-carrageenan, between
0.003-0.07 wt % of phycocyanin from a spirulina-extract and a
chelating agent, wherein the weight ratio of .lamda.-carrageenan to
phycocyanin from a spirulina-extract is between 1:1-100:1.
Preferably the chelating is EDTA. More preferably the chelating
agent EDTA is present in an amount between 1 and 100 ppm.
[0076] In an embodiment the liquid foodstuff is a beverage
comprising between 0.01-0.5 wt % of .lamda.-carrageenan,
phycocyanin from a spirulina-extract and a chelating agent and
wherein the weight ratio of .lamda.-carrageenan to phycocyanin from
a spirulina-extract is between 2:1-50:1. Preferably the chelating
agent is EDTA present in an amount from 10-50 ppm. Preferably the
pH is between 4.6-10.
[0077] Process
[0078] Liquid foodstuffs are typically transported and stored
before being consumed. To assure that these liquid foodstuffs do
not spoil and are safe for consumption, thermal processes such as
retort or UHT (Ultra High Temperature) are used for shelf stable
low acid foodstuff. Pasteurization (73.degree. C. for 15 seconds)
or High Pressure Processing (300-600 MPa) may be used for
refrigerated short shelf life products (60 days). Liquid foodstuffs
can also be preserved through water activity and alcohol content.
Preservatives may be added for additional microbiological
control.
[0079] A considerable blue color loss is observed upon UHT
processing (137.degree. C. for 6 seconds) for low acid
foodstuffs.
[0080] The multi-sulphated carrageenans and/or pectin in
combination with a chelator such as EDTA showed a surprisingly high
increase in color retention and extended shelf life after thermal
treatment and/or use of preservatives.
[0081] The liquid foodstuff can be obtained according to a process
comprising the following steps: [0082] a) Adding and dissolving the
multi-sulphated carrageenans and/or pectin in water and mixing
until the multi-sulphated carrageenans and/or pectin are dissolved,
as determined by visual observation. [0083] b) Adding the
phycocyanin containing spirulina-extract, and mixing until
dissolved at a pH of at least 5, preferably between 5 and 10;
[0084] c) Adding a chelating agent, such as EDTA; [0085] d)
Optionally adding a sweetener, flavor, vitamin, mineral, salt,
buffer, juice, or other beverage components; [0086] e) Optionally
adding other pigments, such as anthocyanins; [0087] f) Adding
solvents to obtain the total volume, such as water and alcohol;
[0088] g) Treating the mixture of a)-f) by either thermally
processing the liquid to at least 65.degree. C. and filling it into
a container; filling the liquid into a container and thermally
processing the filled package to at least 100.degree. C. or no
thermal treatment and cold filling into a container and controlling
by formulation (alcohol or water activity), addition of
preservatives or high pressure.
[0089] The multi-sulphated carrageenans can be hydrolyzed with heat
and acid to reduce viscosity and neutralized to a pH of at least
5.0 before blending with phycocyanin in step a).
[0090] The phycocyanins, multi-sulphated carrageenans and/or
pectin, and EDTA are preferably dissolved in the liquid feedstock.
Dissolved means that the phycocyanins, carrageenans and/or pectin,
and EDTA stay in the solution. The solution is clear to the eye and
no sediment or floating particles are observed.
[0091] Multi-sulphated carrageenan and/or pectin can be dry blended
with a non-acidic carbohydrate such as sucrose or maltodextrin to
improve dissolution of the multi-sulphated carrageenan.
[0092] It is noted that the invention relates to all possible
combinations of features described herein, preferred in particular
are those combinations of features that are present in the claims.
It will therefore be appreciated that all combinations of features
relating to the composition according to the invention; all
combinations of features relating to the process according to the
invention and all combinations of features relating to the
composition according to the invention and features relating to the
process according to the invention are described herein.
[0093] It is further noted that the term `comprising` does not
exclude the presence of other elements. However, it is also to be
understood that a description on a product/composition comprising
certain components also discloses a product/composition consisting
of these components. The product/composition consisting of these
components may be advantageous in that it offers a simpler, more
economical process for the preparation of the product/composition.
Similarly, it is also to be understood that a description on a
process comprising certain steps also discloses a process
consisting of these steps. The process consisting of these steps
may be advantageous in that it offers a simpler, more economical
process.
[0094] The invention is now elucidated by way of the following
examples, without however being limited thereto.
REFERENCES
[0095] Buchweitz, M. (2016). 17-Natural Solutions for Blue Colors
in Food. In R. Carle, & R. M. Schweiggert (Eds.), Handbook on
Natural Pigments in Food and Beverages (pp. 355-384): Woodhead
Publishing. [0096] Dewi, E. N. et al. 2017, "Physical
characteristics of phycocyanin from Spirulina microcapsules using
different coating materials with freeze drying method", IOP Conf.
Ser.: Earth Environ. Sci. 55. [0097] Selig, M. J., et at, (2017),
"Protection of blue color in a spirulina derived phycocyanin
extract from proteolytic and thermal degradation via complexation
with beet-pectin", Food Hydrocolloids. [0098] Singh, Satish K. and
Jacobsson, Sven P. (1996). Kinetics of acid hydrolysis of
.kappa.-carrageenan as determined by molecular weight
(SEC-MALLSRI), gel breaking strength, and viscosity measurements.
Carbohydrate Polymers, 23, 89-103. [0099] Yoshikawa & Belay
(2008) Yoshikawa, N., & Belay, A. (2008). Single-Laboratory
Validation of a Method for the Determination of c-Phycocyanin and
Allophycocyanin in Spirulina (Arthrospira) Supplements and Raw
Materials by Spectrophotometry. Journal of AOAC International, 91,
524-529.
Examples
Experiment 1
Experiment 1.1
[0100] Various hydrocolloids were investigated to determine which
ones prevented phycocyanin aggregation in an aqueous solution near
the isoelectric point of spirulina proteins, .about.pH 4.0. Seven
hydrocolloids were tested at a dosage level of 0.05 wt % in a 7 wt
% sucrose solution at pH 4.0 with EXBERRY.RTM. Shade Blue Powder
60000002 phycocyanin containing spirulina-extract at 0.5 wt %
(equating to 0.012 wt % phycocyanin). The beverage solution was
thermally processed to 85.degree. C. (microwave) and filled hot
into PET bottles and cooled in a water bath to less than 35.degree.
C.
[0101] Spirulina aggregation occurred in control, K-carrageenan,
and guar gum. No precipitation occurred in lambda carrageenan, iota
carrageenan, HM pectin (citrus and apple), xanthan, and gum acacia.
(see table 1).
TABLE-US-00001 TABLE 1 Aggregation of spirulina proteins in aqueous
solution after thermal processing at pH 4 .lamda.- I- K- HM Pectin
Gum Guar Control carrageenan carrageenan carrageenan (citrus/apple)
Xanthan Acacia Gum Yes No No Yes No No No Yes
Experiment 1.2
[0102] To expand upon learnings in experiment 1.1, seven
hydrocolloids were tested at a dosage level of 0.05 wt % in a 7 wt
% sucrose solution at pH 6.5 with EXBERRY.RTM. Shade Blue Powder
60000002 at 0.5 wt % (equating to 0.0117 wt % phycocyanin) to
determine if color retention is improved. Trisodium citrate or
citric acid was added to the final solution to adjust the pH to
6.5. The beverages were processed to 135.degree. C. for 6 seconds
and filled at ambient temperature into sterile bottles as the
product is low acid and the higher temperatures are needed to
control for pathogens.
[0103] Total phycocyanin content was determined before and after
processing. The Yoshikawa & Belay (2008) method was modified
and instead of diluting the sample to adjust pH to 6.0, the
solution was measured as is at pH 6.5.
[0104] No aggregation or precipitation was observed after thermal
processing at pH 6.5, however, color loss was significant >85%
after thermal processing for the control sample (no hydrocolloid)
and all seven hydrocolloids. In experiment 1.2 no chelating agents
were present.
[0105] Table 2 shows the total phycocyanin content of the control
sample and with the addition of seven different hydrocolloids
before and after processing and whether any precipitation was
observed.
TABLE-US-00002 TABLE 2 Precipitation and total phycocyanin (PC)
content before and after UHT thermal processing at pH 6.5. Total PC
Total PC % Loss (mg/mL) (mg/mL) after Before Heat After Heat
heating Precipitation Control 0.117 0.016 86% No .LAMBDA.- 0.114
0.016 86% No carrageenan I- 0.116 0.015 87% No Carrageenan K- 0.118
0.017 86% No Carrageenan Xanthan Gum 0.117 0.017 86% No Gum Arabic
0.118 0.017 86% No HM Pectin 0.116 0.017 86% No (citrus/apple) HM
Pectin 0.112 0.015 87% No (sugar beet)
Experiment 1.3
[0106] As the hydrocolloids on their own had no impact to color
retention during thermal processing and protein aggregation is not
a concern above a pH of 6.0, an experiment was conducted to
determine the influence of a chelator, in particular Na.sub.2 EDTA
at 30 ppm on its own and in combination with the same seven
hydrocolloids in experiment 1.2.
[0107] Aqueous solutions were prepared containing 7 wt % sucrose
solution, EXBERRY.RTM. Shade Blue Powder 60000002 at 0.5 wt %
(equating to 0.0117 wt % phycocyanin), Na.sub.2 EDTA at 30 ppm, and
seven hydrocolloids at a dosage level of 0.05 wt %. Trisodium
citrate or citric acid was added to the final solution to reach a
pH of 6.5. The beverages were processed to 135.degree. C. for 6
seconds and filled at ambient temperature into sterile bottles as
the product is low acid and the higher temperatures are needed to
control for pathogens.
[0108] Total phycocyanin content was determined before and after
processing and through 6 weeks in accelerated storage at 32.degree.
C. in a hotbox. The Yoshikawa & Belay (2008) method was
modified as solution were measured at pH 6.5 vs. 6.0.
[0109] No aggregation or precipitation was observed after thermal
processing at pH 6.5. The combination of Na.sub.2 EDTA with
.lamda.-carrageenan, -carrageenan, and HM pectin (citrus/apple and
sugar beet) had the best color retention post processing and
through six weeks of storage at 32.degree. C. The color of the
beverage after thermal processing and through six weeks of storage
remained blue for the Na.sub.2 EDTA with .lamda.-carrageenan,
-carrageenan, HM sugar beet pectin, and HM citrus/apple pectin
samples. Surprisingly, the color retention of EDTA with xanthan,
gum arabic, and .kappa.-carrageenan was worse than EDTA alone and
the color was greenish blue.
[0110] Table 3 shows the total phycocyanin content for the control
sample (no hydrocolloid or Na.sub.2 EDTA), the addition of Na.sub.2
EDTA at 30 ppm, and seven hydrocolloids at 0.05 wt % with 30 ppm
Na.sub.2 EDTA before and after processing and through six weeks of
accelerated storage at 32.degree. C.
TABLE-US-00003 TABLE 3 Total phycocyanin (PC) content before and
after UHT thermal processing at pH 6.5 and through 6 weeks at
32.degree. C. storage. Total PC Total PC Total PC Total PC % (g/L)
(g/L) % Loss (g/L) (g/L) Improvement Before Heat After Heat with
heat 3 wk (32.degree. C.) 6 wk (32.degree. C.) vs. Control Control
0.117 0.016 86% 0.012 0.010 Na.sub.2 EDTA 0.118 0.034 71% 0.028
0.024 145% .lamda. Carrageenan + Na.sub.2 EDTA 0.115 0.046 60%
0.035 0.029 197% Carrageenan + Na.sub.2 EDTA 0.118 0.040 66% 0.029
0.025 152% .kappa. Carrageenan + Na.sub.2 EDTA 0.118 0.033 72%
0.026 0.017 72% Xanthan Gum + Na.sub.2 EDTA 0.120 0.029 76% 0.018
0.016 59% Gum Arabic + Na.sub.2 EDTA 0.118 0.032 73% 0.024 0.021
113% HM Pectin (citrus/apple) + Na.sub.2 EDTA 0.116 0.042 64% 0.031
0.026 168% HM Pectin (sugar beet) + Na.sub.2 EDTA 0.112 0.041 63%
0.029 0.025 154%
Experiment 1.4
[0111] To confirm findings from experiment 1.3, a follow-up
experiment was conducted with EXBERRY.RTM. Shade Blue Powder
60000002 at 0.6 wt % (equating to 0.014 wt % phycocyanin) and
sucrose at 7 wt % at pH 6.5 with combinations of Na.sub.2 EDTA at
30 ppm and Ticaloid.RTM. 750.lamda.-carrageenan (TIC GUMS) at 0.05
wt %. The solutions were analyzed by photometer before and after
UHT processing (135.degree. C.) and at week 3 and 6 in 32.degree.
C. hotbox storage. A total of four beverages were tested: control,
.lamda.-carrageenan, Na.sub.2 EDTA, and .lamda.-carrageenan with
Na.sub.2 EDTA at pH 6.5.
[0112] The results (see table 4) confirm that .lamda.-carrageenan
on its own will not improve phycocyanin color stability at neutral
pH. Almost all the phycocyanin blue color in the control and
.lamda.-carrageenan samples were destroyed during thermal
processing and the resulting color was grey. While
.lamda.-carrageenan on its own did not improve color retention, the
combination of .lamda.-carrageenan with Na.sub.2 EDTA was the best
performing sample after thermal processing and through 6 weeks
storage at 32.degree. C. The total phycocyanin content of the
Na.sub.2 EDTA and carrageenan sample was 134% higher than control
sample (see table 4). Na.sub.2 EDTA only sample was also
significantly better than control with a 77% improvement in total
phycocyanin content over control.
[0113] FIG. 1 shows the change in absorbance before and after
thermal processing of the four solutions tested.
TABLE-US-00004 TABLE 4 Total Phycocyanin content through 6 weeks
32.degree. C. storage of UHT processed pH 6.5 beverage solutions
Total Phycocyanin (mg/mL) content % Before After Week Week
Improvement Heat Heat 3 6 from Control Control 0.143 0.017 0.020
0.014 Na2 EDTA 0.140 0.032 0.024 0.024 77% .lamda.-carrageenan
0.136 0.017 0.014 0.011 -22% .lamda.-carrageenan + 0.133 0.047
0.037 0.032 134% Na2 EDTA
Experiment 2
[0114] An experiment was conducted with EXBERRY.RTM. Shade Blue
Powder 60000002 at 0.5 wt % (equating to 0.012 wt % phycocyanin), 7
wt % sucrose, and 15 wt % ethylalcohol. Na.sub.2 EDTA at 30 ppm
and/or Ticaloid.RTM. 750 (TIC GUMS) .lamda.-carrageenan at 0.05 wt
% was added to determine if phycocyanin containing spirulina
concentrate can be stabilized in alcohol containing beverages. The
pH of the solutions were measured and the results were between 7-9.
No acids or buffers were added to the solutions as pH measurements
of alcohol containing solutions are not accurate.
[0115] All formulas were cold filled and the order of addition was
as follows: [0116] 1) Dry blend .lamda.-carrageenan with sugar and
add to 50 wt % of total water (room temperature). Mix until fully
hydrated (if used). [0117] 2) Add phycocyanin containing spirulina
concentrate and mix until dissolved. [0118] 3) Add Na.sub.2 EDTA
(if used). [0119] 4) Add remaining water and alcohol.
[0120] The alcohol containing beverages were analyzed by photometer
within 2 hours (T0) after batching and after 3 and 6 weeks of
accelerated storage at 32.degree. C.
[0121] Photometric measurements at T0, 3 weeks and 6 weeks are in
Table. The results show that spirulina is stabilized by Na.sub.2
EDTA and .lamda.-carrageenan with Na.sub.2 EDTA.
TABLE-US-00005 TABLE 5 Color Stability of Phycocyanin Containing
Spirulina-Extract in 15 wt % Alcohol after 6 weeks at 32.degree. C.
Total Phycocyanin (mg/mL) content in 15 wt % Alcohol Beverage
through 6 weeks @ 32.degree. C. 7 wt % Sucrose and 0.5 wt % EXBERRY
.RTM. Shade Blue Powder 60000002 Total Phycocyanin (mg/mL) T0 3
weeks 6 weeks % Loss 15% ETOH Control 0.092 0.032 0.024 74% 15%
ETOH .lamda.-carrageenan 0.090 0.032 0.023 74% 0.05 wt % 15% ETOH
Na.sub.2 EDTA 30 ppm 0.094 0.052 0.044 53% 15% ETOH
.lamda.-carrageenan + 0.092 0.051 0.043 54% Na.sub.2 EDTA
* * * * *