Further, particular attention is made to minimize injury during and after cutting. In another
embodiment, flowering whole plants are collected by hand and the flowers are then separated for further processing.
Minimization of Environmental Impact and Time Factors of Degradation:
Delivery time of cut plant material to the processing facility and exposure of biomass 5 to sun, high temperature, and other negative environmental factors, should be minimized to
prevent the impact of unwanted degradation processes as described above. For example, in
one embodiment of the การประดิษฐ์นี้, the delivery time for Fabaceae plants for further
processing does not exceed 30 minutes from the time of cutting. In another embodiment,
plants that undergo long distance transport are treated to a post-cutting procedure involving 10 immediately placing the plant biomass into Styrofoam coolers containing bags of frozen gel
packs to help maintain freshness and natural moisture content during overnight delivery to
the processing facility. These procedures were conducted for plant biomass from Lamiaceae
and Moraceae families. Other post-cutting procedures that achieve the results described
above may be used as well. As a nonlimiting example, for many plant species it is beneficial 15 to not only minimize delivery time for processing, but to also keep the cut plant material cool,
by refrigeration if necessary, to prevent and/or minimize unwanted degradation prior to
and/or during processing.
Cleaning Step Prior to Grinding and Maceration:
20 A washing step to remove the soil particles and other debris from plants prior to
further processing is performed once the plant tissue is harvested. The washing is achieved
using a low-pressure rinse for a short duration under conditions to prevent the initiation of
the release of the cell juice from biomass, to cause injury, or to remove valuable components.
For example, ในหนึ่ง รุรุ of the การประดิษฐ์นี้, the washing of the plant biomass 25 was accomplished in less than or equal to 5 minutes with a water pressure of less than or
equal to 1 kg/cm2. Residual water wash did not contain any green or yellow pigments, which
indicates the absence of subsequent injury. The excess water is removed from washed
biomass in order to keep the dry matter content close to natural level.
30 After the plant tissue biomass is harvested, as described above, further processing of
the plant tissue biomass is performed to yield plant cell juice. ในหนึ่ง รุรุ, the harvested plant tissue biomass is subjected to grinding, maceration, and pressing to separate the intracellular content, i.e., the cell juice, and to separate it from the fiber-
enriched press-cake containing predominantly cell walls.
35
An example of a suitable processing protocol involves the steps described below. A hammer mill may be used to grind plants to yield plant tissue particles of a small size in a
short time and without significant increase of biomass temperature. ในหนึ่ง รุรุ, a
modified hammer mill is used to produce the maximum size of macerated plant particles less than or equal to 0.5 centimeters during less than or equal to 10 seconds of treatment, where the increase of biomass temperature is less than or equal to 5° C.
5
Exposure of ground and macerated plant biomass is minimized to prevent the impact of unwanted catabolic processes, as described above. The separation of plant cell juice from fiber-enriched material (or press-cake) is commenced as soon as possible after grinding and
maceration of the plant biomass. The plant biomass is processed in a short time and without 10 significant increase in temperature. ในหนึ่ง รุรุ, immediately after grinding and
maceration, the plant biomass is pressed using a horizontal, continuous screw press
(Compact Press "CP-6", Vincent Corporation, FL). The pressure on the cone is maintained
at level 24 kg/cm2, screw speed is at 12 rpm, and biomass temperature increase is less
than or equal to 5°C.
15
The initial cell juice usually contains small fiber particles, which can absorb valuable cell juice components and also block the hoses and pumps. The above particles should be removed by filtration or low-speed centrifugation. For example, the initial cell juices produced
after the pressing step are filtered through four layers of nylon fabric prior to using the plant 20 cell juice in the methods of the การประดิษฐ์นี้.
Once plant cell juice is separated, the plant cell juice is relatively stable colloidal
dispersion in which organelles represent the dispersed phase and cytoplasm represents the
continuous phase. Cell juice is then treated to a processes involving (1) triggering
25 destabilization of above colloidal dispersion performing a "initiation of ส่วนแยก เมมเบรน
aggregation step" to yield a destabilized cell juice and (2) performing a "ส่วนแยก เมมเบรน
separation step" on destabilized cell juice mixture to yield a ส่วนแยก เมมเบรน (containing
nucleous, or chloroplasts, or chromoplasts, or mitochondria, or combination of thereof) and a
cell juice supernatant. ในหนึ่ง รุรุ, initiation of ส่วนแยก เมมเบรน destabilization is 30 accomplished by subjecting said cell juice to electromagnetic waves at a frequency of 2.45
GHz. In another embodiment the frequency employed is greater than 2.45 GHZ up to 7.0
GHz. After destabilization is achieved, a ส่วนแยก เมมเบรน separation step is performed.
This step includes, for example, separating of destabilized cell juice into the membrane
ส่วนแยก and the cell juice supernatant using separating techniques ที่รวมถึง filtration, or
35 centrifugation, or combination of thereof.
A variety of instruments can be employed in the process of the การประดิษฐ์ in order to generate the electromagnetic waves necessary to destabilize the cell juice: magnetrons, power grid tubes, klystrons, klystrodes, crossed-field amplifier, travelling wave tubes, and
gyrotrons. One such instrument includes, but is not limited to high power magnetron.
5 Conventional and industrial magnetrons operate at a frequency of 915 MHz and 2.45 GHz
and can be employed. However at those frequencies undesirable heat is can be generated
that can denature the cell juice composition. It is therefore advantageous to use
electromagnetic waves operating at frequencies that are substantial higher than the
frequencies of conventional or industrial magnetrons, which allows for destabilization of the 10 cell juice without undesirable denaturing due to heat generation. This frequency is typically
above the frequency of conventional microwave magnetrons, i.e., above 2.45 GHz, in
another embodiment greater than 2.45 GHz and less than about 7 GHz; and in another
embodiment from about 3 to about 6 GHz. During the destabilizing step of the การประดิษฐ์ the
temperature of the cell juice is beneficially maintained below 40°C, in another embodiment 15 below about 35°C, in another embodiment below about 30°C, in another embodiment below
about 25°C, in another embodiment below about 20°C.
The freshly obtained ส่วนแยก เมมเบรน commonly referred to in the art, as "protein-
vitamin concentrate," is a paste having intensive color and specific odor that is plant raw
20 material source specific. The ส่วนแยก เมมเบรน is represented predominantly by
chloroplasts present in the green parts of plant or mostly by chromoplasts present in flowers.
The composition of the ส่วนแยก เมมเบรน includes predominantly phospholipids, membrane
proteins, chlorophyll, nucleus, mitochondria and carotenoids.
25 Process for Preparing ส่วนแยก ไซโตพลาสซึม / ไซโตโซล Derived Cosmetic Compositions
Substantially-Free From ส่วนแยก เมมเบรน
The การประดิษฐ์นี้ also relates to a method for preparing the cytoplasm/ไซโตโซล
ส่วนแยก derived cosmetic compositions substantially-free from ส่วนแยก เมมเบรน exhibiting
antioxidant activity, cell growth stimulation activity, or both antioxidant and cell growth
30 stimulation activities. The method involves providing a cell juice that has been separated
from a fresh plant biomass, as already described above with respect to the Membrane-
Derived Cosmetic Composition. The plant cell juice is then treated under conditions effective
to separate the plant cell juice into a ส่วนแยก เมมเบรน and a ส่วนแยก ไซโตพลาสซึม / ไซโตโซล.
35 The ส่วนแยก ไซโตพลาสซึม / ไซโตโซล can then be optionally further processed under
conditions effective to separate the ส่วนแยก ไซโตพลาสซึม / ไซโตโซล into its component parts, namely the cytoplasm ส่วนแยก and a ไซโตโซล ส่วนแยก. The cytoplasm ส่วนแยก includes
predominantly white soluble proteins; in C3 plants, these proteins largely consist of the
enzyme ribulose-1,5biphosphate carboxylase oxygenase. The ไซโตโซล ส่วนแยก contains low
molecular weight soluble components. ไซโตโซล ส่วนแยก is refined under conditions effective
to yield a cell serum ส่วนแยก having antioxidant activity, cell growth stimulation activity, or 5 both antioxidant and cell growth stimulation activities. The cell serum ส่วนแยก is stabilized
under conditions effective to yield a stable bioactive botanical cosmetic composition
exhibiting antioxidant activity, cell growth stimulation activity, or both antioxidant and cell
growth stimulation activities as described for example, in U.S. Patent Nos. 7,442,391;
7,473,435; 7,537,791; 8,043,635; 8,101,212 and 8,277,852.
The plant cell juice may be obtained from all types of plants. Examples of suitable plants that may