U.S. patent application number 17/611983 was filed with the patent office on 2022-08-18 for composition for increasing bioavailability and promoting absorption of ginsenosides in black ginseng extract.
The applicant listed for this patent is CJ CHEILJEDANG CORPORATION. Invention is credited to Nam Hoon CHO, So Hyeong KIM, Son Woo KIM, Yong Ki SEO, Do Yu SOUNG.
Application Number | 20220257681 17/611983 |
Document ID | / |
Family ID | |
Filed Date | 2022-08-18 |
United States Patent
Application |
20220257681 |
Kind Code |
A1 |
SOUNG; Do Yu ; et
al. |
August 18, 2022 |
COMPOSITION FOR INCREASING BIOAVAILABILITY AND PROMOTING ABSORPTION
OF GINSENOSIDES IN BLACK GINSENG EXTRACT
Abstract
The present invention relates to a composition for increasing
the bioavailability and promoting the absorption of ginsenosides in
a black ginseng extract.
Inventors: |
SOUNG; Do Yu; (Suwon-si,
Gyeonggi-do, KR) ; KIM; Son Woo; (Suwon-si,
Gyeonggi-do, KR) ; KIM; So Hyeong; (Suwon-si,
Gyeonggi-do, KR) ; SEO; Yong Ki; (Suwon-si,
Gyeonggi-do, KR) ; CHO; Nam Hoon; (Suwon-si,
Gyeonggi-do, KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CJ CHEILJEDANG CORPORATION |
Seoul |
|
KR |
|
|
Appl. No.: |
17/611983 |
Filed: |
May 20, 2020 |
PCT Filed: |
May 20, 2020 |
PCT NO: |
PCT/KR2020/006590 |
371 Date: |
November 17, 2021 |
International
Class: |
A61K 36/258 20060101
A61K036/258; A23L 33/105 20060101 A23L033/105; A61K 47/36 20060101
A61K047/36; A61K 47/10 20060101 A61K047/10 |
Foreign Application Data
Date |
Code |
Application Number |
May 20, 2019 |
KR |
10-2019-0058675 |
Claims
1. A composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract, the
composition comprising the black ginseng extract containing
ginsenosides Rk1 and Rg5, wherein the total content of ginsenosides
Rk1 and Rg5 is 20 parts by weight to 90 parts by weight, based on
100 parts by weight of the total content of ginsenosides Rb1, Rb2,
Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and Rh1(s).
2. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
1, wherein when 9 g of the black ginseng extract is administered,
the sum of the AUC (Area under the plasma level-time curve) values
of ginsenosides Rb1, Rb2, Rc, Rd, Rg3(s), Rk1, Rg5, CK, Rh2(s),
PPD(s), Re, Rg1, Rh1(s), and PPT(s) is 500 to 1200 .mu.ghr/ml.
3. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
2, wherein when 9 g of the black ginseng extract is administered,
the AUC (Area under the plasma level-time curve) value of
ginsenoside Rk1 is 20 to 40 .mu.ghr/ml.
4. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
2, wherein when 9 g of the black ginseng extract is administered,
the AUC (Area under the plasma level-time curve) value of
ginsenoside Rg3 is 40 to 80 .mu.ghr/ml.
5. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
2, wherein when 9 g of the black ginseng extract is administered,
the AUC (Area under the plasma level-time curve) value of
ginsenoside Rh2 is 250 to 400 .mu.ghr/ml.
6. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides of claim 1, wherein the Tmax value
of ginsenosides Rb1, Rb2, Rc, Rd, Rg3(s), Rk1, Rg5, CK, Rh2(s),
PPD(s), Re, Rg1, Rh1(s), and PPT(s) in the black ginseng extract is
5 to 10 hours.
7. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
1, wherein the black ginseng extract further comprises acidic
polysaccharides and polyphenols.
8. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
1, wherein the composition is a food or pharmaceutical
composition.
9. The composition for increasing the bioavailability and promoting
the absorption of ginsenosides in a black ginseng extract of claim
1, wherein the black ginseng extract is prepared by a method for
preparing a black ginseng extract enriched with ginsenosides Rk1
and Rg5, the method comprising: a step for steaming ginseng to
prepare black ginseng; a step for extracting the prepared black
ginseng with a solvent; and a step for maturing the obtained black
ginseng extract.
10. The composition for increasing the bioavailability and
promoting the absorption of ginsenosides in a black ginseng extract
of claim 9, wherein the steaming is performed at 70.degree. C. to
120.degree. C. for 3 to 12 times.
11. The composition for increasing the bioavailability and
promoting the absorption of ginsenosides in a black ginseng extract
of claim 9, wherein the steaming is performed under the condition
of 2 hours or more per steaming.
12. The composition for increasing the bioavailability and
promoting the absorption of ginsenosides in a black ginseng extract
of claim 9, wherein the step for maturing the black ginseng extract
is performed at 80.degree. C. or higher.
13. The composition for increasing the bioavailability and
promoting the absorption of ginsenosides in a black ginseng extract
of claim 9, wherein the step for maturing the black ginseng extract
is performed for 3 hours or more.
Description
TECHNICAL FIELD
[0001] The present invention relates to a composition for
increasing the bioavailability and promoting the absorption of
ginsenosides in a black ginseng extract.
BACKGROUND ART
[0002] Ginseng is a perennial herb belonging to the genus Panax of
the Araliaceae family and is one of the herbal medicines used in
oriental medicine. As physiologically active efficacies of general
ginseng, an effect on the central nervous system, an anticancer
effect (Patent Document 1), and the like have been reported. In
particular, about 40 types of ginsenosides have been discovered so
far, and it has been confirmed that they exerted a wide range of
effects on the central nervous system, the endocrine system, the
metabolic system, etc., and showed excellent effects in regulating
body functions, that is, normalizing physiological functions. These
ginsenosides have similar or opposite actions, and it is known that
various effects are exerted by a specific component alone or
through the interaction of several types of components.
[0003] The present inventors studied various raw materials among
processed products derived from ginseng, controlled the conditions
for processing ginseng to prepare black ginseng having an increased
content of ginsenosides present in trace amounts in ginseng, and
confirmed that the bioavailability and absorption rate in the body
of ginsenosides contained in a black ginseng extract were
significantly improved compared to that of red ginseng.
Prior Art Document
[0004] (Patent Document 1) Korean Unexamined Publication No.
2010-0134463
DETAILED DESCRIPTION OF THE INVENTION
Technical Problem
[0005] An object of the present application is to provide a
composition for increasing the bioavailability and promoting the
absorption of ginsenosides in a black ginseng extract.
Technical Solution
[0006] According to an aspect of the present application, there is
provided a method for preparing a black ginseng extract.
[0007] Another aspect of the present application provides a
composition for increasing the bioavailability and promoting the
absorption of ginsenosides in a black ginseng extract.
[0008] Hereinafter, the present application will be described in
detail.
[0009] One embodiment of the present application provides a method
for preparing a black ginseng extract enriched with ginsenosides
Rk1 and Rg5, the method comprising: a step for steaming ginseng to
prepare black ginseng; a step for extracting the prepared black
ginseng with a solvent; and a step for maturing the obtained black
ginseng extract.
[0010] The black ginseng extract is derived from black ginseng
obtained by steaming ginseng multiple times.
[0011] The ginseng may be Korean ginseng (Panax ginseng), Canadian
ginseng (P. quinquefolius), Chinese Ginseng (P. notoginseng),
Bamboo ginseng (P. japonicus), Dwarf ginseng (P. trifolium),
Himalayan ginseng (P. pseudoginseng), Vietnamese ginseng (P.
vietnamensis), American ginseng (Panax quinquefoliurn), or the
like. Ginseng is generally divided into white ginseng and red
ginseng according to processing methods. White ginseng refers to
ginseng obtained by drying unprocessed ginseng gathered right out
of the field, that is, fresh ginseng, as it is. Red ginseng is
ginseng obtained by steaming, drying, and processing fresh ginseng.
The preparation process of red ginseng involves several chemical
changes, such as saponin modification and amino acid changes.
Ginsenoside components that do not exist in ginseng are generated
by the heat applied during the red ginseng preparation process. The
unique active components in red ginseng show cancer-preventive
effect, cancer cell growth-inhibiting effect, blood
pressure-lowering effect, brain nerve cell-protective effect, and
learning ability-improving effect.
[0012] Black ginseng of the present application may be obtained by
steaming nine times any one selected from the group consisting of
ginseng leaves, young shoots, stems, stem shells, roots, root
shells, seeds, fruits, immature fruits, mature fruits, flesh,
pericarp, flowers, androecium, gynoecium, calyxes, stamens, petals,
calyx pieces, carpels, and combinations thereof. The androecium
represents the entire stamens in a flower, and the gynoecium
represents the entire pistils in a flower. The carpel is a
component that makes the pistil of a flower and represents a
deformed form of a leaf, which is generally called a floral leaf.
The ginseng may be, but is not limited to, ginseng roots. Other
parts of ginseng steamed nine times may be mixed.
[0013] The ginsenoside Rk1 is represented by the following
formula:
##STR00001##
[0014] (wherein Glc represents a glycosyl residue.)
[0015] The ginsenoside Rg5 is represented by the following
formula:
##STR00002##
[0016] (wherein Glc represents a glycosyl residue.)
[0017] The steaming may be performed 3 to 12 times at 70.degree. C.
to 120.degree. C.
[0018] The steaming may be performed at 75.degree. C. to
115.degree. C., 80.degree. C. to 110.degree. C., 85.degree. C. to
105.degree. C., 87.degree. C. to 103.degree. C., or 90.degree. C.
to 100.degree. C. When performing steaming under the above
temperature condition, the content of the active ingredient
ginsenosides Rk1 and Rg5 increases, and substances harmful to
health are not generated so that the quality can be optimized.
[0019] The steaming may be performed 3 to 11 times, 4 to 10 times,
5 to 9 times, 6 to 9 times, 7 to 9 times, 8 times, or 9 times. When
performing steaming under the above condition of the number of
steaming, the content of the active ingredient ginsenosides Rk1 and
Rg5 increases, and substances harmful to health are not generated
so that the quality can be optimized. As long as the
bioavailability of the black ginseng extract obtained by applying
the steaming conditions of the present application increases and
the absorption rate is improved, the number of steaming can be
variously applied.
[0020] The steaming time may be 1.5 hours to 6 hours, 2 hours or
more, 2 hours to 6 hours, 2 hours to 5.5 hours, 2 hours to 5 hours,
2 hours to 4.5 hours, 2 hours to 4 hours, 2 hours to 3.5 hours, or
2 hours to 3 hours per steaming. When performing steaming under the
above time condition, the content of the active ingredient
ginsenosides Rk1 and Rg5 increases, and substances harmful to
health are not generated so that the quality can be optimized.
[0021] The method for preparing the black ginseng extract may
further include a step for drying the steamed ginseng each time
completing steaming. Drying may be performed at 25.degree. C. to
60.degree. C., 27.degree. C. to 58.degree. C., or 30.degree. C. to
55.degree. C. In addition, drying may be performed for 10 hours to
30 hours, 12 hours to 28 hours, 12 hours to 26 hours, or 12 hours
to 24 hours.
[0022] As used herein, the term "extract" refers to a material
extracted by any method from a raw material, and is meant to
include, without limitation, all of an extract thus extracted, a
concentrate obtained therefrom, a dried product and powders of the
concentrate.
[0023] The extract may be obtained by extraction from a raw
material or a dried product thereof, and the raw material of the
extract may be a raw material grown or a commercially available raw
material, without limitation.
[0024] When the extract is obtained by extracting the raw material,
conventionally known general extraction methods, such as solvent
extraction, ultrasonic extraction, filtration and reflux
extraction, may be used. The extract may be prepared by using
solvent extraction or reflux extraction. The extraction process may
be repeated several times, after which the extract may be further
subjected to steps such as concentration or freeze-drying. For
example, high concentration extract powders may be prepared by
concentrating the obtained extract under reduced pressure to obtain
a concentrate, freeze-drying the concentrate, and using a grinder.
The extract also includes a fraction obtained by further
fractionating the extract.
[0025] The extract may be obtained by using water, an organic
solvent, or a mixture thereof as an extraction solvent. The organic
solvent may be any one selected from the group consisting of an
alcohol, a lower alcohol having 1 to 4 carbon atoms, hexane
(n-hexane), ether, glycerol, propylene glycol, butylene glycol,
ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl
acetate, benzene, and a mixture thereof.
[0026] When a mixture of water and an organic solvent is used as an
extraction solvent, the mixture of water and an organic solvent may
be a mixture of water and a lower alcohol having 1 to 4 carbon
atoms, and may be a mixture of water and ethanol. In this case, the
solvent may be 20% (v/v) to 90% (v/v) aqueous ethanol solution, 25%
(v/v) to 85% (v/v) aqueous ethanol solution, 30% (v/v) to 80% (v/v)
aqueous ethanol solution, 20% (v/v) to 40% (v/v) aqueous ethanol
solution, 30% (v/v) to 60% (v/v) aqueous ethanol solution, 35%
(v/v) to 75% (v/v) aqueous ethanol solution, 40% (v/v) to 70% (v/v)
aqueous ethanol solution, 45% (v/v) to 65% (v/v) aqueous ethanol
solution, or 50% (v/v) to 80% (v/v) aqueous ethanol solution. When
preparing the black ginseng extract, the extraction solvent may be
added 4 times to 10 times, 5 times to 9 times, or 6 times to 8
times the raw material for extraction, black ginseng. The content
of the active ingredient ginsenosides Rk1 and Rg5 increases during
extraction under the above aqueous ethanol solution condition.
[0027] When preparing the black ginseng extract, extraction may be
performed for 2 hours to 12 hours, 3 hours to 11 hours, 4 hours to
10 hours, 4 hours to 9 hours, or 4 hours to 8 hours. The content of
the active ingredient ginsenosides Rk1 and Rg5 increases during
extraction under the above time condition.
[0028] The black ginseng extract may be prepared by the following
process. The black ginseng obtained by completing the final
steaming and drying may be firstly extracted with 50% (v/v) to 80%
(v/v) aqueous ethanol solution, the residue generated from the
first extraction may be secondly extracted with the aqueous ethanol
solution of the same concentration, the residue generated from the
secondary extraction may be tertiarily extracted with 30% (v/v) to
60% (v/v) aqueous ethanol solution, and the residue generated from
the tertiary extraction may be quaternarily extracted with 20%
(v/v) to 40% (v/v) aqueous ethanol solution. The black ginseng
extract may be each of the extracts obtained in each round and may
be a mixture of the extracts obtained from the first to fourth
extractions. The solvent used for each extraction round may have
various concentrations. That is, to increase the content of the
active ingredient ginsenosides Rk1 and Rg5, the extraction solvent
may be used in various combinations.
[0029] The black ginseng extract may be 55 Brix % to 85 Brix %, 57
Brix % to 83 Brix %, 60 Brix % to 80 Brix %, 62 Brix % to 78 Brix
%, or 65 Brix % to 75 Brix %. When the black ginseng extract is in
the above Brix % concentration range, the quality, such as the
content of active ingredient ginsenosides Rk1 and Rg5, is
optimized, and the viscosity of the black ginseng extract is not
too high, so the desired formulation can be prepared.
[0030] The method for preparing the black ginseng extract may
further include a step for maturing the black ginseng extract after
the extraction step.
[0031] The step for maturing the black ginseng extract may be
performed at 80.degree. C. or higher, 82.degree. C. to 95.degree.
C., 84.degree. C. to 93.degree. C., 86.degree. C. to 91.degree. C.,
or 88.degree. C. to 90.degree. C. The content of ginsenosides Rk1
and Rg5 increases during maturing under the above temperature
condition.
[0032] The step for maturing the black ginseng extract may be
performed for 3 hours or more, 3 hours to 48 hours, 3 hours to 44
hours, 3 hours to 40 hours, 3 hours to 36 hours, 3 hours to 32
hours, 3 hours to 28 hours, or 3 hours to 24 hours. The content of
ginsenosides Rk1 and Rg5 increases during maturing under the above
time condition.
[0033] The total content of ginsenosides Rk1 and Rg5 contained in
the black ginseng extract may be 20 parts by weight to 90 parts by
weight, 25 parts by weight to 85 parts by weight, 30 parts by
weight to 80 parts by weight, 35 parts by weight to 75 parts by
weight, 40 parts by weight to 70 parts by weight, 45 parts by
weight to 65 parts by weight, or 50 parts by weight to 60 parts by
weight, based on 100 parts by weight of the total content of
ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and
Rg1(s). When the total content of ginsenosides Rk1 and Rg5 is
within the above range, the effect of increasing the
bioavailability of the black ginseng extract is excellent.
[0034] In the method for preparing the black ginseng extract,
pressure is not applied during the steaming. Pressure is not
applied separately during the steaming, so harmful substances such
as benzopyrene are not generated. Thus, the black ginseng extract
of the present application has excellent quality in terms of
safety.
[0035] The black ginseng is prepared by steaming ginseng. A black
ginseng extract having a high content of ginsenosides Rk1 and Rg5
can be obtained through extraction of the prepared black ginseng
and maturing. For example, black ginseng was prepared by steaming 3
to 12 times at 70.degree. C. to 120.degree. C., with the condition
of 2 hours or more per steaming. To this black ginseng, 4 to 10
times the weight of the black ginseng of water, a lower alcohol
having 1 to 4 carbon atoms, or a mixed solvent thereof was added,
and the black ginseng was subjected to the extraction for 2 to 12
hours to obtain a black ginseng extract. The black ginseng extract
was concentrated, and the black ginseng concentrate was matured at
80.degree. C. or higher for 3 hours or longer. As a result, the
total content of ginsenosides Rk1 and Rg5, which are known to be
not contained or contained in trace amounts in red ginseng, was 20
parts by weight to 90 parts by weight, based on 100 parts by weight
of the total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1,
Rg3(s), Rk1, Rg5, and Rh1(s), and was significantly higher than
that in the red ginseng extract.
[0036] Another embodiment of the present application provides a
composition for increasing the bioavailability and promoting the
absorption of ginsenosides in a black ginseng extract, the
composition comprising the black ginseng extract containing
ginsenosides Rk1 and Rg5, wherein the total content of ginsenosides
Rk1 and Rg5 is 20 parts by weight to 90 parts by weight, based on
100 parts by weight of the total content of ginsenosides Rb1, Rb2,
Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and Rh1(s).
[0037] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides in the black ginseng
extract of the present application, the total content of
ginsenosides Rk1 and Rg5 may be 25 parts by weight to 85 parts by
weight, 30 parts by weight to 80 parts by weight, 35 parts by
weight to 75 parts by weight, 40 parts by weight to 70 parts by
weight, 45 parts by weight to 65 parts by weight, or 50 parts by
weight to 60 parts by weight, based on 100 parts by weight of the
total content of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Rg3(s),
Rk1, Rg5, and Rh1(s). When the total content of ginsenosides Rk1
and Rg5 is within the above range, the effects of increasing the
bioavailability and promoting the absorption of ginsenosides in the
black ginseng extract are excellent.
[0038] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the total content of ginsenosides Rk1 and Rg5
contained in the black ginseng extract may be 9 mg/g or more, 9
mg/g to 30 mg/g, 9 mg/g to 28 mg/g, 9 mg/g to 26 mg/g, 9 mg/g to 24
mg/g, 9 mg/g to 22 mg/g, 9.1 mg/g to 20 mg/g, 9.1 mg/g to 18 mg/g,
9.2 mg/g to 16 mg/g, 9.2 mg/g to 14 mg/g, or 9.2 mg/g to 12 mg/g.
When the total content of ginsenosides Rk1 and Rg5 is within the
above range, the effects of increasing the bioavailability and
promoting the absorption of ginsenosides in the black ginseng
extract are excellent.
[0039] The term "bioavailability" used concerning the ginsenosides
of the black ginseng extract in the present application refers to
the ratio of the amount of ginsenosides that are absorbed and reach
the systemic circulation in an unchanged form when the black
ginseng extract is administered.
[0040] In the present specification, the term "absorption rate" may
be used interchangeably with the same meaning as
bioavailability.
[0041] In the present application, the bioavailability may be
expressed as the area under the plasma level-time curve (AUC) of
ginsenosides measured when the black ginseng extract was taken, or
may also be expressed by comparing the AUC value when the black
ginseng extract was taken with the AUC value of ginsenosides
measured when the same amount of the red ginseng extract is
taken.
[0042] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, when 9 g of the black ginseng extract is administered,
the sum of the AUC values of ginsenosides Rb1, Rb2, Rc, Rd, Rg3(s),
Rk1, Rg5, CK, Rh2(s), PPD(s), Re, Rg1, Rh1(s), and PPT(s) may be
500 to 1200 .mu.ghr/ml.
[0043] Specifically, in the composition for increasing the
bioavailability and promoting the absorption of ginsenosides of the
present application, when 9 g of the black ginseng extract is
administered, the sum of the AUC values of ginsenosides Rb1, Rb2,
Rc, Rd, Rg3(s), Rk1, Rg5, CK, Rh2(s), PPD(s), Re, Rg1, Rh1(s), and
PPT(s) may be 550 to 1150 .mu.ghr/ml, 600 to 1100 .mu.ghr/ml, 650
to 1050 .mu.ghr/ml, 700 to 1000 .mu.ghr/ml, or 750 to 950
.mu.ghr/ml.
[0044] In addition, in the composition for increasing the
bioavailability and promoting the absorption of ginsenosides of the
present application, when 9 g of the black ginseng extract is
administered, the AUC value of ginsenoside Rk1 may be 20 to 40
.mu.ghr/ml, more specifically 25 to 40 .mu.ghr/ml, 25 to 38
.mu.ghr/ml, 25 to 35 .mu.ghr/ml, or 30 to 35 .mu.ghr/ml.
[0045] In addition, in the composition for increasing the
bioavailability and promoting the absorption of ginsenosides of the
present application, when 9 g of the black ginseng extract is
administered, the AUC value of ginsenoside Rg3 may be 40 to 80
.mu.ghr/ml, more specifically 45 to 75 .mu.ghr/ml, 50 to 70
.mu.ghr/ml, or 55 to 65 .mu.ghr/ml.
[0046] In addition, in the composition for increasing the
bioavailability and promoting the absorption of ginsenosides of the
present application, when 9 g of the black ginseng extract is
administered, the AUC value of ginsenoside Rh2 may be 250 to 400
.mu.ghr/ml, more specifically 270 to 380 .mu.ghr/ml, 290 to 360
.mu.ghr/ml, 300 to 350 .mu.ghr/ml, 310 to 350 .mu.ghr/ml, or 320 to
340 .mu.ghr/ml.
[0047] The term "Tmax value" herein refers to the time at which the
blood concentration reaches the maximum after administration of the
black ginseng extract or red ginseng extract, which is the time
when the absorption of the extract reaches the maximum and is the
moment when the absorption rate and the excretion rate of the
extract are the same. It is an indicator of the absorption rate
when comparing the absorption of extracts.
[0048] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the Tmax value of ginsenosides Rb1, Rb2, Rc, Rd,
Rg3(s), Rk1, Rg5, CK, Rh2(s), PPD(s), Re, Rg1, Rh1(s), and PPT(s)
may be 5 to 10 hours. Specifically, the Tmax value of ginsenosides
in the black ginseng extract may be 5.5 to 9.5 hours, 6.0 to 9.0
hours, 6.5 to 9.0 hours, 6.7 to 8.8 hours, 6.9 to 8.7 hours, or 7.0
to 8.5 hours.
[0049] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the black ginseng extract may further include acidic
polysaccharides and polyphenols.
[0050] The acidic polysaccharides may be contained in an amount of
1 mg/g or more, 1 mg/g to 20 mg/g, 1 mg/g to 18 mg/g, 1 mg/g to 16
mg/g, 1 mg/g to 14 mg/g, 1 mg/g to 12 mg/g, 1.5 mg/g to 10 mg/g, 1
mg/g to 8 mg/g, 2 mg/g to 6 mg/g, 1 mg/g to 4 mg/g, or 2.5 mg/g to
4 mg/g.
[0051] When the acidic polypeptides are contained within the above
range, a synergistic effect between the acidic polypeptides and
ginsenosides Rk1 and Rg5 contained in the black ginseng increases.
Thus, the effects of increasing the bioavailability and promoting
the absorption of ginsenosides in the black ginseng extract are
excellent.
[0052] The polyphenols may be contained in an amount of 15 mg/g or
more, 15 mg/g to 40 mg/g, 15 mg/g to 38 mg/g, 15 mg/g to 36 mg/g,
16 mg/g to 34 mg/g, 17 mg/g to 32 mg/g, 18 mg/g to 30 mg/g, 19 mg/g
to 28 mg/g, 20 mg/g to 26 mg/g, or 20 mg/g to 24 mg/g. When the
polyphenols are contained within the above range, a synergistic
effect between the polyphenols and ginsenosides Rk1 and Rg5
contained in the black ginseng increases. Thus, the effects of
increasing the bioavailability and promoting the absorption of
ginsenosides in the black ginseng extract are excellent.
[0053] The composition for increasing the bioavailability and
promoting the absorption of ginsenosides in the black ginseng
extract of the present application may be a food or pharmaceutical
composition.
[0054] As used herein, the term "administration" means introducing
a predetermined substance to a subject in any suitable way. The
composition of the present application can be administered through
any general route that can reach an in vivo target. The route of
administration of the composition of the present application is not
particularly limited but may be oral or parenteral. The
administration of the present application may be performed 1 to 4
times, 2 to 3 times, or 2 times a day. In addition, the
administration of the present application may be performed for a
period of 4 weeks or more, 8 weeks or more, 4 weeks to 12 weeks, or
8 weeks to 12 weeks.
[0055] The dosage of the black ginseng extract containing
ginsenosides Rk1 and Rg5 contained in the composition for
increasing the bioavailability and promoting the absorption of
ginsenosides in the black ginseng extract depends on the patient's
condition and weight, the degree of the disease, drug form, and
route and period of the administration and may be appropriately
selected depending on the circumstances. For example, the black
ginseng extract containing ginsenosides Rk1 and Rg5 may be
administered at a dose of 0.0001 to 1000 mg/kg, 0.001 to 700 mg/kg,
0.01 to 500 mg/kg, 0.1 to 100 mg/kg, or 0.1 to 100 mg/kg, 0.1 to 10
mg/kg, 0.1 to 8 mg/kg, 0.1 to 6 mg/kg, 0.1 to 5 mg/kg, 0.1 to 4
mg/kg, 0.3 to 3.5 mg/kg per day. The administration may be
performed once or several times per day in a divided manner. The
black ginseng extract containing ginsenosides Rk1 and Rg5 of the
present application is a composition for increasing the
bioavailability and promoting the absorption, and when the
composition is administered in an amount equivalent to the dosage
of an extract containing other ginsenosides in general, it may have
excellent bioavailability of ginsenosides and may have excellent
efficacy of a composition containing other ginsenosides in general.
In addition, even when the composition is administered in an amount
smaller than the dosage of an extract containing other ginsenosides
in general, it may have equivalent, similar, or excellent
bioavailability of ginsenosides and may have equivalent, similar,
or excellent efficacy of a composition containing other
ginsenosides in general. The dosage of the black ginseng extract
containing ginsenosides Rk1 and Rg5 of the present application may
be increased or decreased according to the route of administration,
the degree of the disease, sex, weight, age, or the like.
Accordingly, the above dosage does not limit the scope of the
present application in any way.
[0056] The food composition of the present application may be
provided in the form of a powder, a granule, a tablet, a capsule,
syrup, or a beverage. The food composition may be used with other
food or food additives, in addition to the black ginseng extract
containing ginsenosides Rk1 and Rg5 of the present application,
which is the active ingredient. The mixing amount of the active
ingredient may be determined appropriately depending on the purpose
of use, for example, prevention, health, or therapeutic
treatment.
[0057] The black ginseng extract containing ginsenosides Rk1 and
Rg5 may be contained in the food composition in an amount of 0.001
wt % to 60 wt %, for example, 0.01 wt % to 50 wt %, 0.1 wt % to 45
wt %, 1 wt % to 40 wt %, or 1 wt % to 20 wt %. When the content of
the black ginseng extract in the food composition is less than
0.001 wt %, the effects of the black ginseng extract may not be
sufficiently exhibited. When it exceeds 60 wt %, the effect of the
black ginseng extract may be relatively low relative to the
introduced concentration, but there is no problem in terms of
safety, and thus the black ginseng extract can be used in an amount
greater than the above range.
[0058] There is no particular limitation on the type of the food of
the present application. Examples of the food of the present
application include meat, sausages, bread, chocolate, candies,
snacks, confectioneries, pizza, ramen, other noodles, gums, dairy
products including ice cream, various soups, beverages, teas,
drinks, alcoholic beverages, and vitamin complexes.
[0059] The pharmaceutical composition of the present application
may further include an appropriate carrier, an excipient, or a
diluent, commonly used for preparing the pharmaceutical
composition.
[0060] Examples of the carrier, the excipient, or the diluent that
can be used for the pharmaceutical composition of the present
application include lactose, dextrose, sucrose, sorbitol, mannitol,
xylitol, erythritol, maltitol, starch, acacia rubber, alginate,
gelatin, calcium phosphate, calcium silicate, cellulose,
methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone,
water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc,
magnesium stearate, or mineral oil.
[0061] The pharmaceutical composition of the present invention may
be formulated and used in the form of oral formulations, such as
powders, granules, tablets, capsules, suspensions, emulsions,
syrups, aerosols, or the like, external applications,
suppositories, and sterilized injection solutions, respectively,
according to conventional methods. For example, the pharmaceutical
composition of the present application may be provided in the form
of an aerosol, powders, gels, an ointment, or a drop for easy
application to the mucous membrane. The mucous membrane may be, but
not limited to, a nasal mucosa, an oral mucosa, an airway mucosa,
or an eye mucosa.
[0062] For preparation into formulations, a diluent or an
excipient, such as a commonly used filler, a bulking agent, a
binder, a wetting agent, a disintegrating agent, a surfactant, or
the like may be used. Solid formulations for oral administration
include tablets, pills, powders, granules, capsules, etc. These
solid formulations may be prepared by mixing the compound with at
least one excipient, for example, starch, calcium carbonate,
sucrose, lactose, gelatin, etc.
[0063] Furthermore, in addition to simple excipients, lubricants,
such as magnesium stearate and talc are also used. Liquid
formulations for oral administration include a suspension, a liquid
for internal use, an emulsion, syrup, and the like. In addition to
water and liquid paraffin, which are commonly used simple diluents,
various excipients, for example, a wetting agent, a sweetener, a
flavoring agent, a preserving agent, or the like may be added.
[0064] Formulations for parenteral administration include a
sterilized aqueous solution, a non-aqueous solvent, a suspension,
an emulsion, a lyophilized formulation, and a suppository
formulation. As the non-aqueous solvent or the suspension,
propylene glycol, polyethylene glycol, a plant oil such as olive
oil, an injectable ester such as ethylolate, or the like may be
used. As a base for suppositories, witepsol, macrogol, tween 61,
cacao butter, laurin butter, glycerogelatin, or the like may be
used.
[0065] Solid formulations for oral administration include tablets,
pills, powders, granules, capsules, etc. These solid formulations
are prepared by mixing the pharmaceutical composition of the
present application with at least one excipient, for example,
starch, calcium carbonate, sucrose, lactose, gelatin, etc.
Furthermore, in addition to simple excipients, lubricants, such as
magnesium stearate and talc may also be used.
[0066] Liquid formulations for oral administration include a
suspension, a liquid for internal use, an emulsion, syrup, and the
like. In addition to water and liquid paraffin, which are commonly
used simple diluents, various excipients, for example, a wetting
agent, a sweetener, a flavoring agent, a preserving agent, or the
like may be added.
[0067] Formulations for skin administration may be dusting powders,
emulsions, suspensions, oils, sprays, ointments, cream pastes,
gels, foams, or solutions. The pharmaceutical formulation of the
present application may be an anhydrous ointment, and may contain
paraffin that is suitable for local application and is in a liquid
state at body temperature, particularly, low-viscosity paraffin, or
may contain the natural fats or partially synthesized fats, for
example, coconut fatty acid triglyceride, hydrogenated oil, for
example, hydrogenated peanut oil or caster oil, partial fatty acid
ester of glycerol, for example, glycerol monostearate and
distearate, silicone, for example, polymethylsiloxane, such as
hexamethyldisiloxane or octamethyltrisiloxane. For example, the
pharmaceutical formulation may contain a fatty alcohol, which is
associated with an aqueous cream and serves to increase the
moisture absorption capacity, and sterols, wool wax, other
emulsifying agents and/or other additives.
[0068] The dosage of the black ginseng extract containing
ginsenosides Rk1 and Rg5 contained in the pharmaceutical
composition of the present application is as described above.
[0069] The pharmaceutical composition of the present application
may be administered to mammals such as rats, mice, livestock,
humans, and the like through various routes. Administration may be
performed through, for example, oral, rectal, intravenous,
intramuscular, subcutaneous, endobronchial inhalation, intrauterine
dural, or intracerebroventricular injection.
[0070] The black ginseng extract containing ginsenosides Rk1 and
Rg5 of the present application may be administered to a subject in
need thereof in an effective amount.
[0071] In the present application, the subject in need thereof
means a subject in need of the bioavailability increase and
absorption promotion of ginsenosides in the black ginseng
extract.
[0072] The subject herein may be an animal, including a human or an
animal other than a human.
[0073] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the black ginseng extract may be prepared by the
method for preparing a black ginseng extract enriched with
ginsenosides Rk1 and Rg5, the method comprising: a step for
steaming ginseng to prepare black ginseng; a step for extracting
the prepared black ginseng with a solvent; and a step for maturing
the obtained black ginseng extract.
[0074] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the steaming may be performed 3 to 12 times at
70.degree. C. to 120.degree. C.
[0075] The steaming may be performed at 75.degree. C. to
115.degree. C., 80.degree. C. to 110.degree. C., 85.degree. C. to
105.degree. C., 87.degree. C. to 103.degree. C., or 90.degree. C.
to 100.degree. C. When performing steaming under the above
temperature condition, the content of the active ingredient
ginsenosides Rk1 and Rg5 increases, and substances harmful to
health are not generated so that the quality can be optimized.
[0076] The steaming may be performed 3 to 11 times, 4 to 10 times,
5 to 9 times, 6 to 9 times, 7 to 9 times, 8 times, or 9 times. When
performing steaming under the above condition of the number of
steaming, the content of the active ingredient ginsenosides Rk1 and
Rg5 increases, and substances harmful to health are not generated
so that the quality can be optimized.
[0077] The steaming time may be 1.5 hours to 6 hours, 2 hours or
more, 2 hours to 6 hours, 2 hours to 5.5 hours, 2 hours to 5 hours,
2 hours to 4.5 hours, 2 hours to 4 hours, 2 hours to 3.5 hours, or
2 to 3 hours per steaming. When performing steaming under the above
time condition, the content of the active ingredient ginsenosides
Rk1 and Rg5 increases, and substances harmful to health are not
generated so that the quality can be optimized.
[0078] In the composition for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the step for maturing the black ginseng extract may be
performed at 80.degree. C. or higher, 82.degree. C. to 95.degree.
C., 84.degree. C. to 93.degree. C., 86.degree. C. to 91.degree. C.,
or 88.degree. C. to 90.degree. C. The content of ginsenosides Rk1
and Rg5 increases during maturing under the above temperature
condition.
[0079] The step for maturing the black ginseng extract may be
performed for 3 hours or more, 3 hours to 48 hours, 3 hours to 44
hours, 3 hours to 40 hours, 3 hours to 36 hours, 3 hours to 32
hours, 3 hours to 28 hours, or 3 hours to 24 hours. The content of
ginsenosides Rk1 and Rg5 increases during maturing under the above
time condition.
[0080] The contents described herein concerning the method for
preparing the black ginseng extract, which is one embodiment of the
present application, may be applied to the composition for
increasing the bioavailability and promoting the absorption of
ginsenosides in the black ginseng extract. The contents common
between them will not be described redundantly to avoid undue
complexity of the specification. For example, the contents of the
step for steaming ginseng to prepare black ginseng; the step for
extracting the prepared black ginseng with a solvent; and the step
for maturing the obtained black ginseng extract can be applied to
the composition for increasing the bioavailability and promoting
the absorption of ginsenosides.
[0081] As another aspect of the present application, the present
application provides a method for increasing the bioavailability
and promoting the absorption of ginsenosides in a black ginseng
extract, the method comprising a step for administering to a
subject a composition comprising a black ginseng extract containing
ginsenosides Rk1 and Rg5, wherein the total content of ginsenosides
Rk1 and Rg5 is 20 parts by weight to 90 parts by weight, based on
100 parts by weight of the total content of ginsenosides Rb1, Rb2,
Rc, Rd, Re, Rg1, Rg3(s), Rk1, Rg5, and Rh1(s).
[0082] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, when 9 g of the black ginseng extract is administered,
the AUC (Area under the plasma level-time curve) value of
ginsenosides Rb1, Rb2, Rc, Rd, Rg3(s), Rk1, Rg5, CK, Rh2(s),
PPD(s), Re, Rg1, Rh1(s), and PPT(s) may be 500 to 1200
.mu.ghr/ml.
[0083] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the Tmax of ginsenosides in the black ginseng extract
may be 5 hours to 10 hours.
[0084] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the black ginseng extract may further include acidic
polysaccharides and polyphenols.
[0085] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the composition containing the black ginseng extract
may be a food composition or a pharmaceutical composition.
[0086] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the black ginseng extract may be prepared by the
method for preparing a black ginseng extract enriched with
ginsenosides Rk1 and Rg5, the method comprising: a step for
steaming ginseng to prepare black ginseng; a step for extracting
the prepared black ginseng with a solvent; and a step for maturing
the obtained black ginseng extract.
[0087] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the steaming may be performed 3 to 12 times at
70.degree. C. to 120.degree. C.
[0088] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the steaming may be performed under the condition of 2
hours or more per steaming.
[0089] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the step for maturing the black ginseng extract may be
performed at 80.degree. C. or higher.
[0090] In the method for increasing the bioavailability and
promoting the absorption of ginsenosides of the present
application, the step for maturing the black ginseng extract may be
performed for 3 hours or more.
Effects of the Invention
[0091] The black ginseng extract of the present application has the
effects of increasing bioavailability and promoting the absorption
of ginsenosides and thus, can be usefully used as an active
ingredient.
[0092] However, effects of the present application are not limited
to the above-mentioned effects and other effects not mentioned will
be clearly understood from the following description by a person
skilled in the art.
BRIEF DESCRIPTION OF THE DRAWINGS
[0093] FIGS. 1a to 1c show changes in ginsenosides according to the
steaming time during the preparation of the black ginseng.
[0094] FIGS. 2a to 2c show changes in ginsenosides according to the
maturing temperature and the maturing time of the black ginseng
concentrate.
[0095] FIG. 3 shows a clinical trial design plan for measuring the
bioavailability of ginsenosides in black ginseng concentrate.
BEST MODE FOR CARRYING OUT THE INVENTION
[0096] Hereinafter, the present invention will be described in
detail through Preparation Examples and Experimental examples.
[0097] However, the following Preparation Examples and Experimental
examples merely illustrate the present invention, and the contents
of the present application are not limited by the following
Examples and Experimental examples.
PREPARATION EXAMPLE 1
Preparation of Black Ginseng
[0098] 1-1. Preparation of Red Ginseng (One Steaming-Drying Black
Ginseng)
[0099] Four-year-old fresh ginseng roots were placed into a screw
washer (Samgak fmc, South Korea) and washed once for 3 minutes to
remove soil or foreign substances. The washed fresh ginseng roots
were placed in a steamer and first steamed at 98.degree. C. for 2
hours, excluding the preheating time. The steamer was cooled to a
temperature of 55.degree. C. Then, the steamed fresh ginseng roots
were taken out of the steamer. The internal temperature of a hot
air dryer was maintained at 55.degree. C., and then, the steamed
fresh ginseng roots were added and dried for about 18 hours. The
dried red ginseng was moved to a sun-drying place (a drying
facility under natural light) and dried for over a month. Through
the above process, 930 kg of red ginseng (one steaming-drying) was
prepared.
[0100] 1-2. Preparation of Nine Times Steaming-Nine Times Drying
Black Ginseng
[0101] 930 kg of red ginseng prepared in Preparation Example 1-1
above was steamed under the same steaming and drying conditions as
in Preparation Example 1-1. The above-described steaming and drying
processes were repeatedly performed and were referred to as two
times to nine times steaming-drying depending on the number of
repetitions. As the steaming and drying were repeated, the color
gradually became closer to black. Through the above process, 853.8
kg of nine times steaming-nine times drying black ginseng was
prepared. When preparing the black ginseng extract of Preparation
Example 2 or the black ginseng concentrate of Preparation Example
3, nine times steaming-nine times drying black ginseng was used as
a raw material.
PREPARATION EXAMPLE 2
Preparation of Black Ginseng Extract
[0102] Each of 284.6 kg of the red ginseng prepared in Preparation
Example 1-1 and 284.6 kg of the black ginseng prepared in
Preparation Example 1-2 was taken and pulverized. Then, 70% (v/v)
aqueous ethanol solution corresponding to 6 times the weight of the
pulverized product was added, and the first extraction was
performed for 6 hours at 82.degree. C. The extraction residue
generated from the first extraction was subjected to the second
extraction under the same conditions as in the first extraction.
50% (v/v) aqueous ethanol solution corresponding to 6 times the
weight of the extraction residue generated from the second
extraction was added, and the third extraction was performed for 6
hours at 85.degree. C. 30% (v/v) aqueous ethanol solution
corresponding to 6 times the weight of the extraction residue
generated from the third extraction was added, and the fourth
extraction was performed for 6 hours at 90.degree. C. Purified
water corresponding to 6 times the weight of the extraction residue
generated from the fourth extraction was added, and the fifth
extraction was performed for 6 hours at 95.degree. C.
PREPARATION EXAMPLE 3
Preparation of Black Ginseng Concentrate
[0103] The first to fourth red ginseng extracts and the first to
fourth black ginseng extracts obtained in Preparation Example 2
each were filtered to separate the extracts from the extraction
residues. For each raw material, only the separated extract was
placed in a concentrator and concentrated at 55.degree. C. in a
vacuum concentrator for 4 hours. 231 kg of 70 Brix % red ginseng
concentrate was obtained, and 212 kg of 70 Brix % black ginseng
concentrate was obtained. The obtained red ginseng concentrate and
black ginseng concentrate were used as samples for the following
component analysis and experiments on the bioavailability increase
and absorption rate.
EXPERIMENTAL EXAMPLE 1
Confirmation of Changes in Ginsenosides Content in Black Ginseng
Concentrate According to Steaming Time
[0104] Nine times steaming-drying was carried out in the same
manner as in the steaming conditions of Preparation Example 1-1
except that the steaming time was changed to 1 hour, 2 hours, and 3
hours. The prepared red ginseng was extracted and concentrated
according to Preparation Examples 2 and 3. The ginsenosides content
was measured in the same manner as in Experimental Example 3
below.
[0105] When the steaming time was set to 1 hour, it was confirmed
that ginsenosides Rk1 and Rg5 did not exist even after nine times
steaming-drying as shown in FIG. 1a. Meanwhile, when the steaming
time was set to 2 hours, the content of ginsenosides Rk1 and Rg5
was significantly increased compared to the black ginseng
concentrate prepared with the steaming time of 1 hour (FIG. 1b). In
addition, when the steaming time was set to 3 hours, the content of
ginsenosides Rk1 and Rg5 was increased compared to the case where
the steaming time was set to 2 hours (FIG. 1c).
EXPERIMENTAL EXAMPLE 2
Confirmation of Changes in Ginsenosides Content in Black Ginseng
Concentrate According to Maturing Temperature or Maturing Time
[0106] The black ginseng concentrate obtained in Preparation
Example 3 was placed in a 2 ml vial and matured in a water bath for
8 hours at 70.degree. C., 80.degree. C., and 90.degree. C.,
respectively, and the changes in ginsenosides content over time
were measured. The ginsenosides content was measured in the same
manner as in Experimental Example 3.
[0107] As a result, when maturing the black ginseng concentrate at
70.degree. C., the ginsenosides content did not increase even after
8 hours of maturing (FIG. 2a). Meanwhile, when maturing the black
ginseng concentrate at 70.degree. C., the content of ginsenosides
Rk1 and Rg5 increased over time. Even at 90.degree. C., the content
of ginsenosides Rk1 and Rg5 increased over time as in the case of
maturing at 80.degree. C.
[0108] In addition, as can be seen from FIGS. 2b to 2c, the content
of ginsenosides Rk1 and Rg5 was changed according to the maturing
time. The content of ginsenosides Rk1 and Rg5 increased in the
black ginseng concentrate matured under the condition of 80.degree.
C. or higher for 3 hours or more.
EXPERIMENTAL EXAMPLE 3
Analysis of Ginsenoside Components in Black Ginseng Concentrate
[0109] The analysis of ginsenoside components present in the red
ginseng concentrate or the black ginseng concentrate obtained in
Preparation Example 3 was performed using HPLC. As a sample, a
solution obtained by diluting the black ginseng concentrate
obtained in Preparation Example 3 in methanol at a ratio of 1/30,
followed by filtration through a 0.45 .mu.m filter, was used. HPLC
1260 (DAD) (Agilent, USA) was used. VENUSIL XBP C18 column (4.6
MM.times.250 MM, 5.0 .mu.m) was used, and as mobile phases,
LC-grade water was used as solvent A and acetonitrile as solvent B.
The column temperature was maintained at 35.degree. C. The gradient
conditions were as follows: 0-3.8 min, a flow rate of 1 ml/min,
hold at 30% B; 3.81-4.5 min, a flow rate of 0.5 ml/min, hold at 30%
B; 4.51-6 min, a flow rate of 1.2 ml/min, 30% B increased to 42% B;
7.5-13 min, a flow rate of 0.5 ml/min, hold at 42% B; until 18 min,
a flow rate of 0.5 ml/min, increased to 47.4% B; 18.1-25 min, a
flow rate of 1 ml/min, increased to 55% B; 30-32 min, a flow rate
of 1 ml/min, increased to 60% B; 42-50 min, a flow rate of 1
ml/min, 60% B increased to 90% B; and 50.1-53 min, a flow rate of 1
ml/min, hold at 30% B. The results of the component analysis are
shown below (unit: mg/g).
TABLE-US-00001 TABLE 1 Ginsenoside content Rb1 Rb2 Rc Rd Re Rg1 Rg3
(S) Rk1 Rg5 Rh1 (S) Red ginseng 4.91 2.21 3.23 1.75 3.74 1.29 0.42
0.12 0.46 0.43 Black ginseng 0.83 0.00 0.00 0.00 1.34 0.00 4.12
4.75 4.54 0.94
[0110] As a result of analyzing ginsenosides in the black ginseng
concentrate, it was confirmed that 0.832 mg/g of Rb1, 1.157 mg/g of
Rf, 1.008 mg/g of Rg2(S), 0.265 mg/g of Rg2(R), 0.941 mg/g of
Rg1(S), 0.416 mg/g of Rh1(R), 0.274 mg/g of Rg6, 0.777 mg/g of F4,
0.862 mg/g of Rk3, 1.714 mg/g of Rh4, 4.122 mg/g of Rg3(S), 1.184
mg/g of Rg3(R), 4.747 mg/g of Rk1, and 4.539 mg/g of Rg5
existed.
[0111] When comparing the ginsenosides content contained in the
black ginseng concentrate and the red ginseng concentrate, Rk1 and
Rg5 increased about 40 times and about 10 times, respectively,
compared to those in the red ginseng concentrate.
EXPERIMENTAL EXAMPLE 4
Analysis of Acidic Polysaccharide Components in Black Ginseng
Concentrate
[0112] The analysis of acidic polysaccharide components present in
the red ginseng concentrate or the black ginseng concentrate
obtained in Preparation Example 3 was performed through the
carbazole-sulfuric acid colorimetric method. A sample obtained by
diluting 300 mg of each concentrate obtained in Preparation Example
3 above in 10 ml of distilled water was used. The diluted solution
was heated in boiling water at 90.degree. C. for 3 hours, cooled,
and centrifuged (3000 rpm, 10 minutes). 1 ml of the supernatant was
taken, and 4 ml of ethanol was added to form a white precipitate.
To obtain the white precipitate, the supernatant was removed after
centrifugation (3000 rpm, 10 minutes), 4 ml of distilled water was
added to dissolve the white precipitate, and 1 ml of 1:4 mixture of
n-butanol and CHCl.sub.3 was added, stirred, and then centrifuged
again (3000 rpm, 10 minutes). 4 ml of water extract, which is a
supernatant herein, was taken, and sonication was performed.
[0113] 20 .mu.l of the sonication-completed solution, 80 .mu.l of
distilled water, 50 .mu.l of 0.1% carbazole ethanol reagent
(prepared with 0.125 g of carbazole/100 ml of anhydrous ethanol),
and 600 .mu.l of sulfuric acid, were added to a 2 ml tube and
stirred. Then, 200 .mu.l each was placed in a 96-well plate. The
absorbance was measured at 530 nm, and the content was calculated
using a calibration curve.
[0114] For standard products, galacturonic acid was dissolved in
distilled water to prepare concentrations of 1000, 500, 250, 125
mg/L. 20 .mu.l of the standard solution, 80 .mu.l of distilled
water, 50 .mu.l of 0.1% carbazole ethanol reagent (prepared with
0.125 g of carbazole/100 ml of anhydrous ethanol), and 600 .mu.l of
sulfuric acid were added to a 2 ml tube and stirred. Then, 200
.mu.l each was placed in a 96-well plate. The absorbance was
measured at 530 nm to make a calibration curve, and the
concentration was measured. The acidic polysaccharides content is
shown below (unit: mg/g).
TABLE-US-00002 TABLE 2 Acidic polysaccharides Red ginseng 0.37
Black ginseng 2.63
[0115] As shown in Table 2, the black ginseng concentrate contained
a large amount of acidic polysaccharides compared to the red
ginseng concentrate.
EXPERIMENTAL EXAMPLE 5
Analysis of Polyphenol Components in Black Ginseng Concentrate
[0116] The polyphenols content present in the red ginseng
concentrate and the black ginseng concentrate obtained in
Preparation Example 3 was measured using a Microplate Reader
(Powerwave XS, BioTek, USA). 2 g of sodium carbonate (Sigma 223484,
CAS No. 497-19-8) is taken in a 100 mL constant volume flask.
Distilled water was added to adjust the total volume to 100 mL to
prepare a 2% sodium carbonate reagent. Folin-Ciocalteu's phenol
reagent (Sigma F9252-1L) and distilled water were mixed at a ratio
of 1:1 to prepare 50% Folin-Ciocalteu's phenol reagent, which was
wrapped with aluminum foil to prevent light from being
transmitted.
[0117] As for the test solutions, the red ginseng concentrate and
the black ginseng concentrate obtained in Preparation Example 3
each were diluted with distilled water at a ratio of 1:1. 0.1 mL of
each solution was taken and mixed with 0.1 mL of 50%
Folin-Ciocalteu's phenol reagent and 2 mL of 2% sodium carbonate.
Then, the mixture was allowed to stand in a dark place for 30
minutes, and the absorbance was measured at 750 nm. The standard
solution was prepared by taking 0.4 g of gallic acid (Sigma G7384,
CAS No. 149-91-7) in a 100 mL constant volume flask, adding
distilled water to adjust the total volume to 100 mL, and diluting
the solution to concentrations of 31.25 ppm, 62.5 ppm, 125 ppm, 250
ppm, and 500 ppm. The standard solution was mixed with the reagents
in the same manner as the test solutions. Then, the mixture was
allowed to stand in a dark place for 30 minutes, and the absorbance
was measured at 750 nm.
[0118] After absorbance measurement, a calibration curve was
prepared with the absorbance of the standard solution as the
abscissa axis and the concentration of the standard solution as the
ordinate axis. The total polyphenol content of the red ginseng
concentrate and the black ginseng concentrate obtained in
Preparation Example 3 was calculated using Equation 1 below.
Total polyphenol content (mg/mL)=(A.times.B.times.C)/D [Equation
1]
[0119] * A: Total amount of test solution (mL), B: Dilution factor,
C: Total polyphenol concentration in the test solution (mg/mL), D:
Collected sample amount (mL)
[0120] The polyphenols content is shown below (unit: mg/g).
TABLE-US-00003 TABLE 3 Polyphenols Red ginseng 10.4 Black ginseng
20.8
[0121] As shown in Table 3, the black ginseng concentrate contained
a large amount of polyphenols compared to the red ginseng
concentrate.
[0122] Summary of Experimental Examples 3 to 5
[0123] The black ginseng concentrate prepared in Preparation
Example 3 had ginsenosides Rk1 and Rg5, acidic polysaccharides, and
polyphenols content higher than that of the red ginseng concentrate
prepared in Preparation Example 3. Accordingly, the black ginseng
concentrate of Preparation Example 3 was named as a standardized
black ginseng concentrate. The above standardized black ginseng
concentrate was used for measuring the bioavailability of
ginsenosides.
EXPERIMENTAL EXAMPLE 6
Measurement of Bioavailability of Ginsenosides in Black Ginseng
Concentrate
[0124] In Experimental Example 6, after single oral administration
of the black ginseng concentrate and the red ginseng concentrate to
healthy male volunteers, the pharmacokinetic (bioavailability)
characteristics of ginsenosides were compared and evaluated.
[0125] The clinical trial design plan was conducted in a
randomized, open, two-way crossover trial form (see FIG. 3).
Subjects judged suitable for the clinical trial were selected by
conducting screening tests, such as inquiry, physical examinations,
and clinical laboratory tests within three weeks (-21 d to -1 d)
from the first scheduled health functional food administration date
(1 d) for volunteers only. Selected subjects are randomized to one
of two sequence groups, admitted to the clinical trial center on
the afternoon of -1 d, and asked to be fasted except for drinking
water after dinner. On the morning of the health functional food
administration day (1 d), the black ginseng concentrate or the red
ginseng concentrate was orally administered according to the
assigned sequence group, followed by the clinical trial according
to a set schedule. Subjects were discharged on the morning of 2 d.
A drug-free period is at least 14 days after the first
administration. On the afternoon of 14 d, the day before the second
period, subjects were hospitalized again, maintained fasting after
dinner, and received the black ginseng concentrate or the red
ginseng concentrate on the morning of 15 d. After that, the
clinical trial was conducted according to the schedule and subjects
were discharged on the morning of 16 d.
[0126] The rationale for setting the dosage of the black ginseng is
as follows. Ginsenosides have been approved by the Korean Ministry
of Food and Drug Safety for their functionalities of improving
immunity, relieving fatigue, improving blood circulation through
inhibition of platelet aggregation, improving memory, and helping
antioxidant effect. To be recognized for these five functional
contents, the daily intake of the sum of ginsenosides Rb1, Rg1, and
Rg3, which are indicators of ginsenosides, should be 3 mg-80 mg. In
the clinical trial, the sum of Rb1, Rg1, and Rg3, which are
indicators of ginsenosides, is less than 80 mg in 9 g of the black
ginseng concentrate (T) and 9 g of the red ginseng concentrate (R),
respectively. So, the corresponding dose (9 g) was set as a single
dosage in the clinical trial.
[0127] Blood sampling time points for pharmacokinetic
(bioavailability) evaluation were set to be before dosing and 0,
0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 24 h after dosing
on 1 d/15 d. The method for measuring the sample is as follows.
Regarding the concentrations of the main components of ginsenosides
(Rb1, S-Rg3, S-Rh2, S-PPD, Rg1, S-Rh1, Rg5, Rk1, S-PPT, Rd, Rb2,
Rc, and CK), plasma separated from the collected blood was analyzed
using LC-MS/MS for the maximum plasma concentration (Cmax) and the
area under the blood concentration-time curve until the last
measurable blood sampling time (AUClast) of each ginsenoside.
[0128] 6-1. Assessment of Bioavailability Degree of Fourteen
Ginsenosides in Black Ginseng
[0129] After making two test subjects intake the red ginseng
concentrate and the black ginseng concentrate at 2-week intervals,
the absorption rate of fourteen ginsenosides (Rb1, Rb2, Rc, Rd,
Rg3(s), Rk1, Rg5, CK, Rh2(s), PPD(s), Re, Rg1, Rh1(s), and PPT(s))
was analyzed. As a result of analyzing the absorption rate, as
shown in Table 4 below, the absorption rate (based on the AUC) of
the black ginseng concentrate was: 1.8 times for the sum of 14
ginsenosides; 15.7 times for Rg3; and 11.7 times for Rh2, compared
to the red ginseng concentrate, and each increased. In particular,
in the red ginseng concentrate-ingested group, Rk1 absorption was
0, whereas, in the black ginseng concentrate-ingested group, 32
.mu.ghr/ml of the uptake rate for Rk1 (based on the AUC) was
measured. The total absorption rate of all ginsenosides was 456
.mu.ghr/ml in the red ginseng group and 770 .mu.ghr/ml in the black
ginseng group, increasing by 1.7 times. In addition, in terms of
the absorption speed, the time to reach the maximum total
ginsenosides content in the red ginseng concentrate was 11 hours
after administration (Tmax 11 hr), whereas the time to reach the
maximum total ginsenosides content in the black ginseng concentrate
was 8.5 hours after administration (Tmax 8.5 hr). So, the
absorption speed was also excellent in the black ginseng
concentrate.
TABLE-US-00004 TABLE 4 Component Rb1 Rb2 Rc Rd Re Rg1 Rg3 Rk1 Rg5
Rh2 PPD CK Rh1 PPT AUC Tmax Red 87 35 34 13 ND ND 4 ND -- 28 17 230
ND 8 456 11 ginseng Black 25 12 8 7 ND ND 60 32 -- 332 39 246 7 4
770 8.5 ginseng Multiplying -3.4 -3 -4.6 -1.9 15.7 32 11.7 2.3 1.1
7 -2.1 1.7 factor
PREPARATION EXAMPLE 4
Preparation of Foods
[0130] 4-1. Preparation of Wheat Flour Foods
[0131] 0.5 to 5.0 parts by weight of the black ginseng extract of
the present invention was added to wheat flour, and this mixture
was used to prepare bread, cakes, cookies, crackers, and
noodles.
[0132] 4-2. Preparation of Soups and Gravies
[0133] 0.2 to 5.0 parts by weight of the black ginseng extract of
the present invention was added to soups and gravies to prepare
meat-processed products for health improvement, noodle-style soups,
and gravies.
[0134] 4-3. Preparation of Ground Beef
[0135] 10 parts by weight of the black ginseng extract of the
present invention was added to ground beef to prepare ground beef
for health improvement.
[0136] 4-4. Preparation of Dairy Products
[0137] 5 to 10 parts by weight of the black ginseng extract of the
present invention was added to milk, and various dairy products
such as butter and ice cream were prepared using the milk.
[0138] 4-5. Preparation of Dry Cereal
[0139] Brown rice, barley, glutinous rice, and adlay were
pregelatinized by a conventional method, dried, roasted, and
prepared into powders by a pulverizer to have a particle size of 60
mesh.
[0140] Black beans, black sesame, and perilla seeds were also
steamed and dried by a conventional method, roasted, and prepared
into powders by a pulverizer to have a particle size of 60
mesh.
[0141] The black ginseng extract of the present invention was
concentrated under reduced pressure, sprayed, and dried by a hot
air dryer. The resulting dry product was prepared into powders by a
pulverizer to have a particle size of 60 mesh.
[0142] The thus-prepared grains, seeds and nuts, and the black
ginseng extract of the present invention were mixed in the
following ratio to prepare dry cereal:
[0143] grains (brown rice (30 parts by weight), adlay (15 parts by
weight), and barley (20 parts by weight)), seeds and nuts (perilla
seeds (7 parts by weight), black beans (8 parts by weight), black
sesame (7 parts by weight), the black ginseng extract of the
present invention (3 parts by weight), Ganoderma lucidum (0.5 part
by weight), and Rehmannia glutinosa (0.5 part by weight).
[0144] 4-6. Preparation of Health Drinks
[0145] 5 g of the black ginseng extract of the present invention
was homogeneously mixed with minor ingredients, such as high
fructose corn syrup (0.5%), oligosaccharide (2%), sugar (2%), salt
(0.5%), and water (75%). The mixture was sterilized instantly and
packaged in small containers, such as glass bottles, pet bottles,
etc., to prepare health drinks.
[0146] 4-7. Preparation of Vegetable Juice
[0147] 5 g of the black ginseng extract of the present invention
was added to 1,000 ml of tomato or carrot juice to prepare
vegetable juice.
[0148] 4-8. Preparation of Fruit Juice
[0149] 1 g of the black ginseng extract of the present invention
was added to 1,000 ml of apple or grape juice to prepare fruit
juice.
PREPARATION EXAMPLE 5
Preparation of Pharmaceutical Compositions
[0150] 5-1. Preparation of Powders
[0151] The black ginseng extract of the present invention 2 g
[0152] Lactose 1 g
[0153] The above ingredients were mixed and filled in an airtight
pouch to prepare powders.
[0154] 5-2. Preparation of Tablets
[0155] The black ginseng extract of the present invention 100
mg
[0156] Corn starch 100 mg
[0157] Lactose 100 mg
[0158] Magnesium stearate 2 mg
[0159] The above noted ingredients were mixed and tabletted
according to a conventional tablet preparation method to provide
tablets.
[0160] 5-3. Preparation of Capsules
[0161] The black ginseng extract of the present invention 100
mg
[0162] Corn starch 100 mg
[0163] Lactose 100 mg
[0164] Magnesium stearate 2 mg
[0165] The above ingredients were mixed and filled in a gelatin
capsule according to a conventional method for preparing capsules
to prepare capsules.
[0166] 5-4. Preparation of Pills
[0167] The black ginseng extract of the present invention 1 g
[0168] Lactose 1.5 g
[0169] Glyserine 1 g
[0170] Xylitol 0.5 g
[0171] The above ingredients were mixed and prepared into a pill
according to a conventional method in such a manner that one pill
has a weight of 4 g.
[0172] 5-5. Preparation of Granules
[0173] The black ginseng extract of the present invention 150
mg
[0174] Soybean extract 50 mg
[0175] Glucose 200 mg
[0176] Starch 600 mg
[0177] The above ingredients were mixed and 100 mg of 30% ethanol
was added thereto, followed by drying at 60.degree. C. After
formation of granules, the granules were filled into packaging.
* * * * *