U.S. patent application number 16/888782 was filed with the patent office on 2020-12-10 for method for simultaneously extracting lycopene and citrulline from watermelon.
The applicant listed for this patent is ZHENGZHOU FRUIT RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES. Invention is credited to Dalei CHEN, Zhonggao JIAO, Hui LIU, Jiechao LIU, Zhenzhen LV, Wenbo YANG, Chunling ZHANG, Qiang ZHANG.
Application Number | 20200385323 16/888782 |
Document ID | / |
Family ID | 1000005100632 |
Filed Date | 2020-12-10 |
United States Patent
Application |
20200385323 |
Kind Code |
A1 |
JIAO; Zhonggao ; et
al. |
December 10, 2020 |
METHOD FOR SIMULTANEOUSLY EXTRACTING LYCOPENE AND CITRULLINE FROM
WATERMELON
Abstract
A method for simultaneously extracting Lycopene and Citrulline
from a watermelon includes: separating the rind and the pulp of the
watermelon, preprocessing the rind, and using the preprocessed rind
to extract the Citrulline; subjecting the pulp to biological
enzymolysis and filtering, centrifuging a filtrate, using a
precipitate and a filter residue obtained after the centrifuging to
extract the Lycopene, and using a supernatant obtained after the
centrifuging to extract the Citrulline. By using the method for
synchronously extracting Lycopene and Citrulline from the
watermelon of the present invention, about 0.5 kg of Lycopene (6%
content) and more than 1.2 kg of Citrulline which are worthy of
nearly ten thousand yuan can be extracted from each ton of
imperfect watermelons, the economic benefit of each ton of
watermelons can be increased by more than 5000 yuan after
extraction costs are deducted, and the method is high in economic
benefits.
Inventors: |
JIAO; Zhonggao; (Zhengzhou,
CN) ; LIU; Jiechao; (Zhengzhou, CN) ; ZHANG;
Chunling; (Zhengzhou, CN) ; ZHANG; Qiang;
(Zhengzhou, CN) ; LIU; Hui; (Zhengzhou, CN)
; LV; Zhenzhen; (Zhengzhou, CN) ; YANG; Wenbo;
(Zhengzhou, CN) ; CHEN; Dalei; (Zhengzhou,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ZHENGZHOU FRUIT RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL
SCIENCES |
Zhengzhou |
|
CN |
|
|
Family ID: |
1000005100632 |
Appl. No.: |
16/888782 |
Filed: |
May 31, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
PCT/CN2019/076477 |
Feb 28, 2019 |
|
|
|
16888782 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07C 7/005 20130101;
C07C 273/189 20130101 |
International
Class: |
C07C 7/00 20060101
C07C007/00; C07C 273/18 20060101 C07C273/18 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 30, 2018 |
CN |
201811002394.9 |
Claims
1. A method for synchronously extracting Lycopene and Citrulline
from watermelon, which is characterized in that: separate the rind
and pulp of watermelon; the Citrulline is extracted from the rind
after pretreatment; process the pulp with biological enzymolysis,
then filter to obtain filter residue and filtrate; the filtrate is
centrifuged to obtain precipitate and supernatant, the precipitate
is mixed with the filter residue above to use to extract Lycopene,
and the supernatant is used to extract Citrulline.
2. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 1, wherein the process of
biological enzymolysis the amount of biological enzyme added is
0.1-0.3% (m/m), with the temperature maintaining at 40-50.degree.
C., and then performing 1-2 hours of enzymolysis.
3. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 2, wherein the biological enzyme
is at least one of pectinase, cellulase and protease.
4. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 1, wherein the biological
enzymatic treatment of pulp is: watermelon pulp is beaten after the
seeds are removed; the slurry is added with 0.3% (m/m) of
biological enzymes to perform enzymatic hydrolysis for 1.5 h at
45.degree. C.; Then the enzymatic slurry is filtered to obtain
filter residue and filtrate, and the filtrate is centrifuged to
collect the precipitate; after being lyophilized, the precipitate
and filter residue which are combined are ground into powder for
extraction of Lycopene; the supernatant after centrifugation is
used for extraction of Citrulline.
5. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 4, wherein the method for
extracting Lycopene is: (1) ultrasound-assisted organic solvent
leaching: add ethyl acetate to the powder ground from the filter
residue and the precipitate at a ratio of 1:4-1:8 (kg/L) and mix
evenly with the temperature 40-50.degree. C., ultrasonic output
power 80-120 W, extraction time 30-90 min, and extraction times
twice to obtain liquid of crude Lycopene extract; (2) concentrated
under reduced pressure: the liquid of crude Lycopene extract is
concentrated under reduced pressure at 40.degree. C., and the
solvent is recovered to obtain a crude Lycopenet; (3) purification:
Anhydrous ethanol is added to the crude Lycopene at a
material-liquid ratio of 1:3 (kg/L) to embathe for twice; And then
a Lycopene powder is obtained after removing the anhydrous
ethanol.
6. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 1, wherein the pretreatment of
the watermelon rind is: remove the outer skin of watermelon rind,
and then squeeze the rind to obtain the filter residue and rind
juice; The filter residue is subjected to ultrasound-enzyme
assisted solvent leaching to obtain an extract solution; And then
the extract solution is mixed with rind juice and the supernatant
which is obtained by centrifuging the pulp to obtain a pretreatment
material for extracting Citrulline.
7. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 1, wherein the pretreatment of
the watermelon rind is: remove the outer skin of watermelon rind,
and ground the rind into powder after being dried; the powder is
subjected to ultrasound-enzyme assisted solvent leaching and
filtered to obtain a primary extraction solution; the filter
residue is subjected to ultrasound-enzyme assisted solvent
leaching, and then filtered to obtain a secondary extraction
solution; the secondary extraction solution is mixed with the
primary extraction solution and the supernatant after
centrifugation of pulps, to obtain a pretreatment material for
extracting Citrulline.
8. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 6, wherein the
ultrasonic-enzyme-assisted solvent leaching is: water is added to
the pretreated material at a material-liquid ratio of 1:10-1:20
(kg/L), and then 0.1% (m/m) of pectinase and 0.1% (m/m) of
cellulose are also added to the above pretreated material; adjust
pH to 4.0, ultrasonic power to 100-140 W, extraction temperature to
30-50.degree. C., extraction time to 60-120 min, and extraction
times to twice.
9. The method for synchronously extracting Lycopene and Citrulline
from watermelon according to claim 6, wherein the method for
extracting Citrulline is: (1) microbial fermentation: 5% (v/v)
yeast is added to the crude extract of Citrulline to ferment for 24
h to remove sugar; (2) ion-exchange resin purification: the crude
extract of Citrulline after microbial fermentation is filtered to
remove yeasts, and added to HD-8 activated wet resin to adsorb
Citrulline, and then eluted with 0.5 mol/L ammonia solution for 2
BV/h of the elution rate; (3) macroporous adsorption resin
discoloration: mix XAD-761 activated wet resin with Citrulline
eluent at 1:20 (g/mL), and perform static adsorption on condition
of 100-150 r/min for 2 h at room temperature; (4) crystallization
and recrystallization: a. The crude Citrulline extract is
concentrated under vacuum to a soluble solid content of
.gtoreq.30%, the pH of the concentrated solution is adjusted to
5.97, and the Citrulline crystals are precipitated at 4.degree. C.;
The precipitate is centrifuged, washed, and dried to obtain a white
powdered solid of Citrulline crystals. b. Dissolve the Citrulline
powder of step (a) with water, adjust the pH to 5.97, add 0.1%
Citrulline crystals as seed crystals, precipitate the Citrulline
crystals at 4.degree. C., and centrifuge to separate the
precipitates; c. Repeat step (b) 1-2 times, centrifuge the
precipitate, wash, and dry to obtain a refined solid powder of
Citrulline.
10. (canceled)
Description
BACKGROUND OF THE INVENTION
1. Technical Field
[0001] The invention belongs to the technical field of natural
substance extraction, and particularly relates to a method for
synchronously extracting Lycopene and Citrulline from
watermelon.
2. Description of Related Art
[0002] Watermelons are widely planted in China, with short growth
cycles and high yields. The annual planting area is about 2 million
hectares, with a total output of 68 million tons, accounting for
58.99% and 71.45% of the world's total watermelon cultivation area
and total output respectively, and both ranking first in the world.
Due to the strong seasonality of watermelon planting, the market is
concentrated, and it is not easy to store. The imbalance between
supply and demand has caused watermelon prices to fall sharply or
even to overstock, causing a lot of decay. In addition, the
defective melons in the planting process are not suitable for fresh
sales, and often can only be rotten in the ground or sold extremely
cheaply, resulting in a waste of resources. The key to solving
these problems lies in vigorously developing deep processing of
watermelon and increasing the added value of watermelon.
[0003] In addition to fat and cholesterol, watermelon contains a
large amount of glucose, malic acid, fructose, amino acids,
Lycopene and rich vitamin C and other substances. It is a
nutrient-rich, pure, and safe food.
[0004] At present, there are extraction methods for functional
components such as Citrulline and Lycopene in watermelon. However,
these functional components exist in different parts of watermelon.
Extracting a certain natural substance singly can not maximize the
making full use of the nutritional value of watermelons. It has
caused waste of resources and increased production costs.
BRIEF SUMMARY OF THE INVENTION
[0005] In view of the problems existing in the prior art, the
object of the present invention is to provide a method for
synchronously extracting Lycopene and Citrulline from watermelon,
which maximize the making full use of the nutritional value of
watermelon, reduce production costs and obtain higher economic
benefits.
[0006] The technical solution adopted by the invention to fulfill
the above objective is as follows:
[0007] A method for synchronously extracting Lycopene and
Citrulline from watermelon:
[0008] separate the rind and pulp of watermelon; the Citrulline is
extracted from the rind after pretreatment; process the pulp with
biological enzymolysis, then filter to obtain filter residue and
filtrate; the filtrate is centrifuged to obtain precipitate and
supernatant, the precipitate is mixed with the filter residue above
to use to extract Lycopene, and the supernatant is used to extract
Citrulline.
[0009] Wherein the process of biological enzymolysis, the amount of
biological enzyme added is 0.1-0.3% (m/m), with the temperature
maintaining at 40-50.degree. C., and then performing 1-2 hours of
enzymolysis.
[0010] The biological enzyme is at least one of pectinase,
cellulase, and protease.
[0011] In one embodiment, the biological enzymatic treatment of the
pulp is:
[0012] Watermelon pulp is beaten after the seeds are removed. The
slurry is added with 0.3% (m/m) of biological enzymes to perform
enzymatic hydrolysis for 1.5 h at 45.degree. C. Then the enzymatic
slurry is filtered to obtain filter residue and filtrate, and the
filtrate is centrifuged to collect the precipitate. After being
lyophilized, the precipitate and filter residue which were combined
are ground into powder for extraction of Lycopene; the supernatant
after centrifugation is used for extraction of Citrulline.
[0013] The method for extracting Lycopene is:
[0014] (1) Ultrasound-assisted organic solvent leaching: add ethyl
acetate to the powder ground from the filter residue and the
precipitate at a ratio of 1:4-1:8 (kg/L) and mix evenly with the
temperature 40-50.degree. C., ultrasonic output power 80-120 W,
extraction time 30-90 min, and extraction times twice to obtain
liquid of crude Lycopene extract;
[0015] (2) Concentrated under reduced pressure: the liquid of crude
Lycopene extract is concentrated under reduced pressure at
40.degree. C., and the solvent is recovered to obtain a crude
Lycopene;
[0016] (3) Purification: Anhydrous ethanol is added to the crude
Lycopene at a material-liquid ratio of 1:3 (kg/L) to embathe for
twice. And then a Lycopene powder is obtained after removing the
anhydrous ethanol.
[0017] In one embodiment, the pretreatment of the watermelon rind
is:
[0018] Remove the outer skin of watermelon rind, and then squeeze
the rind to obtain the filter residue and rind juice. The filter
residue is subjected to ultrasound-enzyme assisted solvent leaching
to obtain an extract solution. And then the extract solution is
mixed with the rind juice and the supernatant which is obtained by
centrifuging the pulp to obtain a pretreatment material for
extracting Citrulline.
[0019] In one embodiment, the pretreatment of the watermelon rind
is:
[0020] Remove the outer skin of watermelon rind, and ground the
rind into powder after being dried. The powder is subjected to
ultrasound-enzyme assisted solvent leaching and filtered to obtain
a primary extraction solution. The filter residue is subjected to
ultrasound-enzyme assisted solvent leaching, and then filtered to
obtain a secondary extraction solution. The secondary extraction
solution is mixed with the primary extraction solution and the
supernatant after centrifugation of pulps, to obtain a pretreatment
material for extracting Citrulline.
[0021] In any of the preceding embodiments, the ultrasound-enzyme
assisted solvent leaching is:
[0022] Water is added to the pretreated material at a
material-liquid ratio of 1:10-1:20 (kg/L), and then 0.1% (m/m) of
pectinase and 0.1% (m/m) of cellulose are also added to the above
pretreated material. Adjust pH to 4.0, ultrasonic power to 100-140
W, extraction temperature to 30-50.degree. C., extraction time to
60-120 min, and extraction times to twice.
[0023] In one embodiment, the extraction method of Citrulline
is:
[0024] (1) Microbial fermentation: 5% (v/v) yeast is added to the
pretreated material to ferment for 24 hours to remove sugar.
[0025] (2) Ion-exchange resin purification: the pretreated material
after microbial fermentation is filtered to remove yeasts, and
added to HD-8 activated wet resin to adsorb Citrulline, and then
eluted with 0.5 mol/L ammonia solution for 2 BV/h of the elution
rate to obtain Citrulline eluent.
[0026] (3) Macroporous adsorption resin discoloration: mix XAD-761
activated wet resin with Citrulline eluent at 1:20 (g/mL), and
perform static adsorption on condition of 100-150 r/min for 2 h at
room temperature.
[0027] (4) Crystallization and recrystallization:
[0028] a. The crude Citrulline extract is concentrated under vacuum
to a soluble solid content .gtoreq.30%, the pH of the concentrated
solution is adjusted to 5.97, and the Citrulline crystals are
precipitated at 4.degree. C. The precipitate is centrifuged,
washed, and dried to obtain a white powdered solid of Citrulline
crystals.
[0029] b. Dissolve the Citrulline powder of step (a) with water,
adjust the pH to 5.97, add 0.1% (m/m) Citrulline crystals as seed
crystals, precipitate the Citrulline crystals at 4.degree. C., and
centrifuge to separate the precipitates.
[0030] c. Repeat step (b) 1-2 times, centrifuge the precipitate,
wash, and dry to obtain a refined solid powder of Citrulline.
[0031] The Lycopene and/or Citrulline which be prepared by the
method described above.
[0032] The Lycopene prepared by the above method can apply to the
preparation of foods for anti-oxidation, anti-aging, anti-hypoxia
response, prevention and treatment of cancer, and protection of
cardiovascular.
[0033] The Citrulline prepared by the above method can apply to the
preparation of foods for enhancing immunity, anti-oxidation,
improving exercise function, protecting cardiovascular and
cerebrovascular, and improving male sexual function.
[0034] Based on the above scheme, the food is a health product.
[0035] The strain number of yeast is CICC-1012, China Center of
Industrial Culture Collection.
[0036] The pectinase, cellulase, and protease were purchased from
Tianjin Lihua Enzyme Preparation Technology Co., Ltd., and the
enzyme activity was 1000 U/mg.
[0037] Advantages of the technical solution of the present
invention:
[0038] The synchronous extraction method of watermelon Lycopene and
Citrulline achieves sufficient and efficient utilization of
watermelon resources. The extraction rate of Lycopene and
Citrulline is high, and the purity of the extracted Citrulline is
high. The extracted Lycopene and Citrulline meet the relevant
sanitary requirements and product quality standards, and can be
used as raw materials in the food and health products industry, and
has natural advantages of safety.
[0039] The present invention uses a combination of ultrasonic,
enzymatic and other auxiliary extraction methods to ensure
efficient extraction under low temperature conditions. The
conditions are mild and the operation is simple and safe.
[0040] Using the synchronous extraction method of watermelon
Lycopene and Citrulline of the present invention, about 0.5 kg (6%
content) of Lycopene and more than 1.2 kg of Citrulline can be
extracted per ton of inferior watermelon, with a value of nearly
10,000 yuan. After deducting the cost of extraction, each ton of
watermelon can still increase the economic benefit by more than
5,000 yuan, which has higher economic benefits.
DETAILED DESCRIPTION OF THE INVENTION
[0041] Terms used in the present invention have the meanings
generally understood by those of ordinary skill in the art unless
otherwise specified.
[0042] The present invention will be described in further detail
with reference to specific embodiments and with reference to data.
The following examples are only for the purpose of illustrating the
present invention, and are not intended to limit the scope of the
present invention in any way.
Embodiment 1
[0043] A method for synchronously extracting Lycopene and
Citrulline from watermelon, the steps are as follows:
[0044] Isolated the rind and pulp of watermelon;
[0045] Treatment of Pulp:
[0046] Watermelon pulp was beaten after the seeds were removed. The
slurry was added with 0.1% pectinase, 0.1% cellulase, and 0.1%
protease to perform enzymatic hydrolysis for 1.5 h at 45.degree. C.
Then the enzymatic slurry was filtered to obtain filter residue and
filtrate; the filtrate is centrifuged (4000 r/min, room
temperature, 10-20 min) to obtain precipitate and supernatant.
After being lyophilized, the precipitate and filter residue which
have been already combined were ground into powder for extraction
of Lycopene; the supernatant after centrifugation would be used for
extraction of Citrulline.
[0047] The extraction method of Lycopene is:
[0048] (1) Ultrasound-assisted organic solvent leaching: added
ethyl acetate to the powder ground from the filter residue and the
precipitate at a ratio of 1:6 (kg/L) and mixed evenly with the
temperature 45.degree. C., ultrasonic output power 100 W,
extraction time 30 min, and extraction times twice to obtain liquid
of crude Lycopene extract;
[0049] (2) Concentrated under reduced pressure: the liquid of crude
Lycopene extract was concentrated under reduced pressure at
40.degree. C., and the solvent was recovered to obtain a crude
Lycopenet;
[0050] (3) Purification: Anhydrous ethanol was added to the crude
Lycopene at a material-liquid ratio of 1:3 (kg/L) to embathe for
twice. And then a Lycopene powder was obtained after removing the
anhydrous ethanol.
[0051] Treatment of Watermelon Rind:
[0052] Remove the outer skin of watermelon rind, and then squeeze
the rind to obtain the filter residue and rind juice.
[0053] Water was added to the filter residue at a material-liquid
ratio of 1:10 (kg/L), and then 0.1% (m/m) of pectinase and 0.1%
(m/m) of cellulose were also added to the above filter residue.
Adjusted pH to 4.0, ultrasonic power to 100 W, extraction
temperature to 50.degree. C., extraction time to 90 min, and
extraction times to twice. And then the combined extract was
obtained.
[0054] The above combined extract was mixed with the supernatant
which was obtained by centrifugation of the rind juice and pulp to
obtain a pretreatment material for extracting Citrulline.
[0055] Extraction Method of Citrulline:
[0056] (1) Microbial fermentation: 5% (v/v) yeast was added to the
pretreated material to ferment for 24 hours to remove sugar.
[0057] (2) Ion-exchange resin purification: the pretreated material
after microbial fermentation was filtered to remove yeasts, and
added to HD-8 activated wet resin to perform the dynamic exchange
adsorption of Citrulline, and then eluted with 0.5 mol/L ammonia
solution for 2 BV/h of the elution rate.
[0058] The dynamic exchange adsorption is that 180 g of activated
wet resin was packed in a glass chromatography column with an inner
diameter of 26 mm and a height of 400 mm, and the Citrulline
extract solution after fermentation by microorganisms was passed
through the resin column at a speed of 2 BV/h to adsorb
Citrulline.
[0059] (3) Macroporous adsorption resin discoloration: mixed
XAD-761 activated wet resin with Citrulline eluent at 1:20 (g/mL),
and performed static adsorption on condition of 100-150 r/min for 2
h at room temperature.
[0060] (4) Crystallization and recrystallization:
[0061] a. The crude Citrulline extract was concentrated under
vacuum to a soluble solid content of .gtoreq.30%, the pH of the
concentrated solution was adjusted to 5.97, and the Citrulline
crystals were precipitated at 4.degree. C. The precipitate was
centrifuged, washed, and dried to obtain a white powdered solid of
Citrulline crystals.
[0062] b. Dissolved the Citrulline powder of step (a) with water,
adjusted the pH to 5.97, added 0.1% Citrulline crystals as seed
crystals, precipitated the Citrulline crystals at 4.degree. C., and
centrifuged to separate the precipitates.
[0063] c. Repeated step (b) 1-2 times, centrifuged the precipitate,
washed, and dried to obtain a refined solid powder of
Citrulline.
[0064] After testing, the extraction rate of Lycopene was 91.57%;
the extraction rate of Citrulline was 93.43%, and the purity of
Citrulline was 99.35%.
Embodiment 2
[0065] A method for synchronously extracting Lycopene and
Citrulline from watermelon, the steps are as follows:
[0066] Isolated the rind and pulp of watermelon;
[0067] Treatment of Pulp:
[0068] Watermelon pulp was beaten after the seeds were removed. The
slurry was added with 0.1% pectinase, 0.1% cellulase, and 0.1%
protease to perform enzymatic hydrolysis for 1.5 h at 45.degree. C.
Then the enzymatic slurry was filtered to obtain filter residue and
filtrate; the filtrate is centrifuged (4000 r/min, room
temperature, 10-20 min) to obtain precipitate and supernatant.
After being lyophilized, the precipitate and filter residue which
have been already combined were ground into powder for extraction
of Lycopene; the supernatant after centrifugation would be used for
extraction of Citrulline.
[0069] The extraction method of Lycopene is:
[0070] (1) Ultrasound-assisted organic solvent leaching: added
ethyl acetate to the powder ground from the filter residue and the
precipitate at a ratio of 1:6 (kg/L) and mixed evenly with the
temperature 45.degree. C., ultrasonic output power 100 W,
extraction time 30 min, and extraction times twice to obtain liquid
of crude Lycopene extract;
[0071] (2) Concentrated under reduced pressure: the liquid of crude
Lycopene extract was concentrated under reduced pressure at
40.degree. C., and the solvent was recovered to obtain a crude
Lycopenet;
[0072] (3) Purification: Anhydrous ethanol was added to the crude
Lycopene at a material-liquid ratio of 1:3 (kg/L) to embathe for
twice. And then a Lycopene powder was obtained after removing the
anhydrous ethanol.
[0073] Treatment of Watermelon Rind:
[0074] The watermelon rind which the outer skin was removed was
ground into powder after being dried;
[0075] Water was added to the powder at a material-liquid ratio of
1:10 (kg/L), then 0.1% (m/m) of pectinase and 0.1% (m/m) of
cellulose were also added to the above powder. Adjusted pH to 4.0,
ultrasonic power to 100 W, extraction temperature to 50.degree. C.,
and extraction time to 90 min, and a primary extraction solution
was obtained after being filtered. Water was added to the filter
residue at a material-liquid ratio of 1:10 (kg/L), then 0.1% (m/m)
of pectinase and 0.1% (m/m) of cellulose were also added to the
above filter residue. Adjusted pH to 4.0, ultrasonic power to 100
W, extraction temperature to 50.degree. C., and extraction time to
90 min, and a secondary extraction solution was obtained after
being filtered.
[0076] The secondary extraction solution was mixed with the primary
extraction solution and the supernatant after centrifugation of
pulp to obtain a pretreatment material for extracting
Citrulline.
[0077] Extraction Method of Citrulline:
[0078] (1) Microbial fermentation: 5% (v/v) yeast was added to the
pretreated material to ferment for 24 hours to remove sugar.
[0079] (2) Ion-exchange resin purification: the pretreated material
after microbial fermentation was filtered to remove yeasts, and
added to HD-8 activated wet resin to perform the dynamic exchange
adsorption of Citrulline, and then eluted with 0.5 mol/L ammonia
solution for 2 BV/h of the elution rate.
[0080] The dynamic exchange adsorption is that 180 g of activated
wet resin was packed in a glass chromatography column with an inner
diameter of 26 mm and a height of 400 mm, and the Citrulline
extract solution after fermentation by microorganisms was passed
through the resin column at a speed of 2 BV/h to adsorb
Citrulline.
[0081] (3) Macroporous adsorption resin discoloration: mixed
XAD-761 activated wet resin with Citrulline eluent at 1:20 (g/mL),
and performed static adsorption on condition of 100-150 r/min for 2
h at room temperature.
[0082] (4) Crystallization and recrystallization:
[0083] a. The crude Citrulline extract was concentrated under
vacuum to a soluble solid content of .gtoreq.30%, the pH of the
concentrated solution was adjusted to 5.97, and the Citrulline
crystals were precipitated at 4.degree. C. The precipitate was
centrifuged, washed, and dried to obtain a white powdered solid of
Citrulline crystals.
[0084] b. Dissolved the Citrulline powder of step (a) with water,
adjusted the pH to 5.97, add 0.1% Citrulline crystals as seed
crystals, precipitated the Citrulline crystals at 4.degree. C., and
centrifuged to separate the precipitates.
[0085] c. Repeat step (b) 1-2 times, centrifuged the precipitate,
washed, and dried to obtain a refined solid powder of
Citrulline.
[0086] After testing, the extraction rate of Lycopene was 91.76%;
the extraction rate of Citrulline was 92.98%, and the purity of
Citrulline was 98.77%.
[0087] 1. Analysis and test results of Lycopene and Citrulline
extract of the present invention
[0088] (1) Analysis of Lycopene
[0089] The hygienic indicators such as heavy metals and
microorganisms in the watermelon Lycopene extract extracted in
Example 1 were tested and compared with the quality standards of
commercially available Lycopene oleoresin products. The results are
shown in Table 1.
TABLE-US-00001 TABLE 1 Hygienic indicators of watermelon Lycopene
extract Commercial product Test item Unit Test result standard
Arsenic mg/kg 5.3 .times. 10.sup.-2 .ltoreq.5 Lead mg/kg Not
detected .ltoreq.10 Total bacteria CFU/g <10 .ltoreq.1000 Yeast
and mold CFU/g <10 .ltoreq.100 E. coli MPN/100 g <30
.ltoreq.40
[0090] From the test results in Table 1, it can be known that the
indexes of heavy metals and microorganisms of the watermelon
Lycopene extract extracted by the method of the present invention
meet the relevant sanitary requirements and product quality
standards.
[0091] (2) Analysis of Citrulline
[0092] The physicochemical and hygienic indicators of the
Citrulline extract extracted in Example 1 were analyzed and
compared with the quality standards of commercially available
Citrulline products. The results are shown in Table 2.
TABLE-US-00002 TABLE 2 Physicochemical and hygienic indicators of
Citrulline extract Commercial product Test item Unit Test result
standard Citrulline content % 92.48 -- Ammonium content % <0.02
.ltoreq.0.02 (NH.sub.4) Chloride % <0.02 .ltoreq.0.02 Slsfate
(SO.sub.4) % <0.02 .ltoreq.0.02 Iron mg/kg <10 .ltoreq.10
Arsenic mg/kg 2.7 .times. 10.sup.-2 .ltoreq.1 Lead mg/kg Not
detected .ltoreq.10
[0093] From the test results in Table 2, it can be known that the
heavy metal and microbe indexes of the Citrulline extract extracted
by the method of the present invention meet the relevant health
requirements and product quality standards.
[0094] Economic Benefits
[0095] The current price of Lycopene on the Chinese market is about
2,000 yuan/kg (6% content), and Citrulline extracted from plants
can be sold for about $1,000/kg in the United States. By using the
method of the present invention, about 0.5 kg (6% content) of
Lycopene and more than 1.2 kg of Citrulline can be extracted per
ton of inferior watermelon, and the value is nearly 10,000 yuan.
After deducting the extraction cost, the economic benefit can still
increase by more than 5000 yuan per ton of watermelon. Based on the
annual output of 70 million tons of watermelon in China, if 10% of
the processing capacity can be achieved, the output value can reach
more than 60 billion yuan. If the watermelon with higher content of
Lycopene and Citrulline is used as the raw material, the extraction
yield and economic benefits will be higher.
[0096] The above embodiments are only used to illustrate the
technical solutions of the present invention, but not to limit
them. Although the present invention has been described in detail
with reference to the foregoing embodiments, the technical
solutions described in the embodiments are still possibly modified,
or some of the technical features are equivalently replaced by
those skilled in the art by referring to the foregoing. These
modifications or replacements do not depart from the spirit and
scope of the technical solutions claimed in the present
invention.
* * * * *