U.S. patent application number 09/935320 was filed with the patent office on 2002-08-15 for bioprocess for the high-yield production of food flavor-acceptable jasmonic acid and methyl jasmonate, novel jasmonic acid isomer produced thereby and uses thereof.
Invention is credited to Blocker, Robert W., Farbood, Mohamad I., Hagedorn, Myrna, Kim, Augustine Yonghwi, Kossiakoff, Nicolas, McLean, Lynda B., McLean, Michael P., Sprecker, Mark A..
Application Number | 20020110881 09/935320 |
Document ID | / |
Family ID | 23858566 |
Filed Date | 2002-08-15 |
United States Patent
Application |
20020110881 |
Kind Code |
A1 |
Farbood, Mohamad I. ; et
al. |
August 15, 2002 |
Bioprocess for the high-yield production of food flavor-acceptable
jasmonic acid and methyl jasmonate, novel jasmonic acid isomer
produced thereby and uses thereof
Abstract
Described is a bioprocess for the high-yield production of food
flavor-acceptable jasmonic acid and methyl jasmonate, as well as a
novel jasmonic acid isomer produced thereby and organoleptic uses
thereof. The process yields at least 5% of the "cis" isomer defined
according the structure: 1 (wherein R is hydrogen or methyl) or at
least 5% of the "cis" isomer defined according to the structure: 2
(wherein R is hydrogen or methyl). Compositions containing at least
98% of the isomer having the structure: 3 with an optical rotation
(.alpha..sub.D.sup.2) of +58.degree. are novel. Compositions
containing at least 98% of the isomer having the structure: 4 with
an optical rotation (.alpha..sub.D.sup.20) of +58.degree. are also
novel. The process of our invention comprises the cultivation under
aerobic condition of one or more specific strain of Diplodia
gossypina in a nutrient medium followed either by (1) isolation of
the jasmonic acid product or (2) esterification of the jasmonic
acid to form methyl jasmonate followed by the isolation of the
methyl jasmonate and novel products produced by such process.
Inventors: |
Farbood, Mohamad I.; (State
College, PA) ; Blocker, Robert W.; (Brick, NJ)
; McLean, Lynda B.; (Matawan, NJ) ; Sprecker, Mark
A.; (Sea Bright, NJ) ; McLean, Michael P.;
(Matawan, NJ) ; Kossiakoff, Nicolas; (Les Arcs sur
Argens, FR) ; Kim, Augustine Yonghwi; (Morganville,
NJ) ; Hagedorn, Myrna; (Woodland Park, CO) |
Correspondence
Address: |
Joseph F. Leightner, Esq.
INTERNATIONAL FLAVORS & FRAGRANCES INC.
521 West 57th Street
New York
NY
10019
US
|
Family ID: |
23858566 |
Appl. No.: |
09/935320 |
Filed: |
August 22, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
09935320 |
Aug 22, 2001 |
|
|
|
09468134 |
Dec 21, 1999 |
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|
Current U.S.
Class: |
435/148 ;
435/252.3; 562/503 |
Current CPC
Class: |
C07C 59/82 20130101;
C12P 7/62 20130101; C12N 1/14 20130101; A23L 27/203 20160801; C12P
7/40 20130101; A23L 27/24 20160801; C11B 9/003 20130101; C07C
2601/08 20170501; C07C 69/738 20130101; A24B 15/34 20130101 |
Class at
Publication: |
435/148 ;
435/252.3; 562/503 |
International
Class: |
C12P 007/26; C12N
001/20; C07C 061/16 |
Claims
What is claimed is:
1. A process for the manufacture of food flavor acceptable jasmonic
acid in relatively high yield of from 1-2 grams per liter,
containing at least 5% of a "cis" isomer having a structure
selected from the group consisting of: 132and a maximum of 95% by
weight of at least one of the isomers having a structure selected
from the group consisting of: 133which comprises the steps of (i)
cultivation under aerobic conditions of a strain of Diplodia
gossypina organism selected from the group consisting of: (a)
Diplodia gossypina ATCC 10936; (b) Diplodia gossypina ATCC 20575;
(c) Diplodia gossypina NRRL 25011; and (d) Diplodia gossypina ATCC
36037 in a nutrient medium containing an assimilable source of
carbon and an assimilable source of nitrogen followed by (ii)
isolation of the jasmonic acid product from the culture medium.
2. The process of claim 1 wherein the compound
10-oxo-trans-8-decenoic acid having the structure: 134is intimately
admixed in the nutrient medium in an amount of from about 0.7 up to
about 10 ppm by weight of the nutrient medium prior to the
cultivation step.
3. The process of claim 1 wherein the cultivation takes place under
aeration conditions at an aeration rate in the range of from about
0.5 V/V/M up to about 1.0 V/V/M.
4. The process of claim 2 wherein the cultivation takes place under
aeration conditions at an aeration rate in the range of from about
0.5 V/V/M up to about 1.0 V/V/M.
5. The process of claim 1 wherein the nutrient medium contains from
about 6.times.10.sup.-3 up to about 10.5.times.10.sup.-3 gram
moles/liter of [Mg.sup.++].
6. The process of claim 5 wherein the source of the [Mg.sup.++] is
MgSO.sub.4.7H.sub.2O and the range of concentrations of said
MgSO.sub.4.7H.sub.2O is from about 1.5 up to about 2.5 grams per
liter.
7. The process of claim 1 wherein the strain of Diplodia gossypina
is Diplodia gossypina ATCC 10936.
8. The process of claim 7 wherein the compound
10-oxo-trans-8-decenoic acid having the structure: 135is intimately
admixed in the nutrient medium in an amount of from about 0.7 up to
about 10 ppm by weight of the nutrient medium prior to the
cultivation step.
9. The process of claim 7 wherein the cultivation takes place under
aeration conditions at an aeration rate in the range of from about
0.5 V/V/M up to about 1.0 V/V/M.
10. The process of claim 7 wherein the nutrient medium contains
from about 6.times.10.sup.-3 up to about 10.5.times.10.sup.-3 gram
moles/liter of [Mg.sup.++].
11. The process of claim 2 wherein the nutrient medium contains
from about 6.times.10.sup.-3 up to about 10.5.times.10.sup.-3 gram
moles/liter of [Mg.sup.++].
12. The process of claim 1 wherein methyl jasmonate containing at
least 25% by weight of the isomer having the structure: 136is
produced from the jasmonic acid by means of reaction of the
resulting jasmonic acid with methyl alcohol according to the
reaction: 137wherein the final isolation comprises the steps of:
(a) extraction of the jasmonic acid from the fermentation broth
with an extraction solvent to form a jasmonic acid extract; (b)
concentration of the resulting jasmonic acid extract, whereby the
extraction solvent is stripped; (c) esterification of the resulting
jasmonic acid with methyl alcohol, whereby methyl jasmonate is
formed according to the reaction: 138(d) concentration of the
resulting methyl jasmonate.
13. The process of claim 1 wherein the jasmonic acid produced is at
least 98% of the optical isomer having the structure: 139having an
optical rotation (.alpha..sub.D.sup.20) of +58.degree. and
comprising the additional steps of (iii) extraction of the
resulting jasmonic acid product with a solvent in order to form a
jasmonic acid product solvent extract; (iv) concentrating the
resulting extract by means of solvent evaporation, thereby forming
a concentrate; (v) fractionating the concentrate with silica gel in
order to effect isomer separation; and (vi) collecting said 98%
optical cis isomer.
14. The process of claim 1 carried out in liquid surface
culture.
15. The process of claim 1 carried out in submerged liquid
culture.
16. The process of claim 15 wherein the cultivation takes place
under aeration conditions at an aeration rate in the range of from
about 0.5 V/VIM up to about 1.0 V/V/M.
17. The process of claim 1 wherein the assimilable carbon source is
glucose.
18. The process of claim 1 wherein the assimilable nitrogen source
is sodium nitrate.
19. The process of claim 1 wherein the incubation time is from
about 3 up to about 15 days.
20. The process of claim 1 wherein the pH range is from about 4.5
up to about 9.
21. The process of claim 20 wherein the pH of the fermentation
broth is 6.
22. The process of claim 1 wherein the temperature range of
reaction is from about 20.degree. C. up to about 35.degree. C.
23. The process of claim 1 wherein the temperature range of the
fermentation is from about 25.degree. C. up to about 32.degree.
C.
24. A jasmonic acid derivative defined according to the structure:
140having an optical rotation (.alpha..sub.D.sup.20) of +58.degree.
produced according to the process of claim 1.
25. A jasmonic acid derivative defined according to the structure:
141wherein R is hydrogen or methyl, having an optical rotation
(.alpha..sub.D.sup.20) of +58.degree. prepared according to the
process of claim 1.
26. A jasmonic acid derivative defined according to the structure:
142having an optical rotation (.alpha..sub.D.sup.20 of +58.degree.
prepared according to the process of claim 13.
27. A process for augmenting, enhancing or imparting an aroma or
taste in or to a consumable material selected from the group
consisting of foodstuffs, chewing gums, beverages and smoking
tobaccos, comprising the step of adding to the consumable material
base an aroma or taste augmenting, enhancing or imparting quantity
and concentration of the product of claim 24.
28. A process for augmenting, enhancing or imparting an aroma in or
to a perfume composition, cologne or perfumed article, comprising
the step of admixing with a perfume base, a cologne base or a
perfumed article base an aroma augmenting, enhancing or imparting
quantity and concentration of the product defined according to
claim 24.
Description
BACKGROUND OF THE INVENTION
[0001] This invention concerns a microbial process for production
of compositions of matter containing isomers of jasmonic acid
having the structure: 5
[0002] which are flavor acceptable as well as isomers having the
structure: 6
[0003] which are also flavor acceptable. This invention is also
concerned with the production of methyl jasmonate isomers from the
aforementioned jasmonic acid isomers.
[0004] Considerable time and effort have been expended by
microbiologists in the search for better processes for the
production of jasmonic acid and methyl jasmonate which are flavor
acceptable isomers, including those isomers defined according to
the structure: 7
[0005] wherein R is methyl or hydrogen. Thus, Broadbent in United
Kingdom Patent Specification No. 1,286,266 published on Aug. 23,
1972 discloses and claims a process for the manufacture of jasmonic
acid which comprises cultivation of the organism Lasiodiplodia
theobromae in a nutrient medium containing an assimilable source of
carbon and an assimilable suurce of nitrogen followed by isolation
of the product from the culture medium. Gunther, et al, German
Democratic Republic Patent DD 279 688 published on Jun. 13,1990
discloses a process for the production of 7-iso-jasmonic acid by
strains of the organism Botryodiplodia theobromae in aerobic
culture. This work is also discussed in the paper Miersch, et al,
Phytochemistry, Volume 26, No. 4, pages 1037-1039, 1987, entitled
"(+)-7-ISO-JASMONIC ACID AND RELATED COMPOUNDS FROM BOTRYODIPLODIA
THEOBROMAE." The organisms, Botryodiplodia theobromae and
Lasiodermea theobromae are synonyms of Diplodia gossypina as
discussed in the paper Jones, MYCOTAXON, Volume VI, No. 1 at pages
24-26, July-September 1997 (title: "THE CURRENT TAXONOMIC STATUS OF
DIPLODIA GOSSYPINA." As confirmed by Husler and Munch in the
article entitled "Microbial Production of Natural Flavors," Volume
63, No. 10, ASM News at pages 551-559:
[0006] "Another plant fatty acid metabolite, jasmonic acid, an
endogenous plant growth regulator with a variety of physiological
functions, is produced by means of a similar metabolic pathway.
After a lipoxygenase produces a hydroperoxide derivative of
linolenic acid, this compound is converted to its allene oxide,
which cyclizes. .beta.-Oxidation and double-bond reduction yields
jasmonic acid. The methylester of jasmonic acid is not only a
volatile plant hormone, possibly involved in interplant
commurnication, but is also an important flavor and fragrance
molecule that imparts a sweet-floral, jasmine-like note . . .
[0007] Otto Miersch and his collaborators at the Institute of Plant
Biochemistry, Halle-Saale, Germany, who were studying fungal plant
pathogens, including Botryodiplodia theobromae, discovered that
such microorganisms produce jasmonic acid. The biosynthetic steps
leading to jasmonic acid in this filamentous fungus are probably
similar to those found in plants. Recently our laboratory, which is
evaluating this strain's capacity for producing jasmonic acid,
found that B. theobromae yields only very low concentrations of
jasmonic acid in liquid culture. Such findings suggest that the
biosynthesis and excretion of jasmonic acid is strictly controlled
during the growth cycle of this fungus on plants in its natural
habitat."
[0008] no bioprocesses for the high yield production of fruit
flavor-acceptable jasmonic acid or methyl jasmonate exist. The
Husler and Munch paper was published in October 1997. ASM News is
published by the American Society for Microbiology.
[0009] Thus, in the flavor and fragrance art, a need has arisen for
the development and efficient high yield production of naturally
occurring jasmonic acid and methyl jasmonate, which have heretofore
been found to be useful and necessary for the creation of flavor
formulations used in augmenting or enhancing the aroma or taste of
foodstuffs, chewing gums, toothpaste, medicinal products, chewing
tobaccos and smoking tobaccos and also useful in augmenting or
enhancing the aroma of perfume compositions, colognes and perfumed
articles (e.g., solid or liquid anionic, cationic, nonionic or
zwitterionic detergents, perfumed polymers, fabric softener
compositions, fabric softener articles, hair preparations, cosmetic
powders and the like).
[0010] Methyl jasmonate (without indicating which isomer) is
disclosed by Arctander, Perfume and Flavor Chemicals at monograph
2093 to have a "Powerful floral-herbaceous, sweet-tenacious odor
representing typical background notes of Jasmin absolute."
Arctander further discloses that this material "is an almost
obvious candidate for work on improved artificial Jasmin absolute."
Acree, et al, J. Agric. Food Chem., 1985, Volume 33, pages 425-427,
discloses that the isomer of methyl jasmonate having the structure:
8
[0011] and having an optical rotation (.alpha..sub.D.sup.20) of +58
has a strong odor, whereas the other stereoisomers of (Z)-methyl
jasmonate are substantially odorless.
[0012] No specific disclosures exist in the prior art showing the
use of the jasmonic acid isomers of our invention defined according
to the structure: 9
[0013] or defined according to the structure: 10
[0014] for use as flavor adjuvants (wherein R is methyl or
hydrogen). Furthermore, nothing in the prior art discloses the
stereoisomer of the compound having the structure: 11
[0015] particularly the stereoisomer having the optical rotation
(.alpha..sub.D.sup.20) of +58.degree..
THE INVENTION
[0016] Our invention relates to a bioprocess for tile high yield
production of food flavor acceptable jasmonic acid and methyl
jasmonate, a novel jasmonic acid isomer produced thereby and uses
thereof. The process of our invention yields at least 5% of the
"cis" isomer defined according to the structure: 12
[0017] (wherein R is hydrogen or methyl) or the "cis" isomer
defined according to the structure: 13
[0018] (wherein R is hydrogen or methyl). Compositions containing
at least 98% of the isomer having the structure: 14
[0019] having an .alpha..sub.D.sup.20 of +58.degree. are novel.
Furthermore, compounds having the structure: 15
[0020] wherein R is methyl or hydrogen having an
.alpha..sub.D.sup.20 of +58.degree. are also novel.
[0021] The process of our invention comprises the cultivation under
aerobic conditions of one or more specific strains of Diplodia
gossypina in a nutrient medium followed by either (1) isolation of
the jasmonic acid or (2) esterification of the jasmonic acid to
form methyl jasmonate followed by isolation of the methyl
jasmonate. More specifically, our process comprises cultivation
under aerobic conditions of a strain of Diplodia gossypina organism
selected from the group consisting of:
[0022] (i) Diplodia gossypina ATCC 10936;
[0023] (ii) Diplodia gossypina ATCC 20575;
[0024] (iii) Diplodia gossypina NRRL 25011; and
[0025] (iv) Diplodia gossypina ATCC 36037
[0026] in a nutrient medium containing an assimilable source of
carbon and an assimilable source of nitrogen followed by isolation
of the jasmonic acid product from the culture medium or followed by
esterification of the jasmonic acid to form methyl jasmonate and
then followed by isolation of the methyl jasmonate product from the
reaction mass.
[0027] In one embodiment of the process of our invention, the
compound, 10-oxo-trans-8-decenoic acid having the structure: 16
[0028] is intimately admixed in the nutrient medium in an amount of
from about 0.7 up to about 10 ppm (parts per million) by weight of
the nutrient medium prior to the cultivation step.
[0029] The 10oxo-trans-8-ecenoic acid having the structure: 17
[0030] is disclosed in U.S. Pat. No. 5,681,738 issued on Oct. 28,
1997 to be a fungal growth hormone to stimulate mycelial growth of
cultivated mushrooms. The specification of U.S. Pat. No. 5,681,738
issued on Oct. 28, 1997 is incorporated by reference herein. The
bioprocess of our invention is shown according to the reactions:
18
[0031] wherein D.G. strain is intended to mean "strain of Diplodia
gossypina."
[0032] The esterification reaction following the bioprocess
reaction of our invention is shown as follows: 19
[0033] Thus, the assimilable carbon source for our invention has
been found to be glucose. The preferred assimilable nitrogen source
is sodium nitrate.
[0034] Preferably, the aeration rate range for carrying out the
cultivation step of our invention is from about 0.5 v/v/m (liters
air per liter fermentation mass per minute) up to about 1.0
v/v/m.
[0035] Furthermore, it is preferable that the culture medium
contain magnesium ion, for example, magnesium ion in the form of
magnesium sulfate heptahydrate having the formula:
MgSO.sub.4.7H.sub.2O
[0036] with the magnesium ion concentration ([Mg.sup.++]) being in
the range of from about 6.times.10.sup.-3 up to about
10.5.times.10.sup.-3 gram moles per liter. Thus, for example, the
concentration range of magnesium sulfate heptahydrate in grams per
liter preferably is in the range of from about 1.5 up to about 2.5
grams per liter of magnesium sulfate heptahydrate.
[0037] The cultivation step of the process of our invention may
take place under liquid surface culture conditions or may take
place under submerged liquid culture conditions. The incubation
time is from about 3 up to about 15 days.
[0038] The pH range during the fermentation is from about 4.5 up to
about 9, with a preferred pH of about 6.
[0039] The temperature for the fermentation reaction may vary
between about 20.degree. C. up to about 35.degree. C., with a
preferred temperature range for the fermentation being from about
25.degree. C. up to about 32.degree. C.
[0040] We have found that the preferred strain of Diplodia
gossypina is the Diplodia gossypina ATCC 10936.
[0041] Along with the desired "cis" jasmonic acid isomers having
the structures: 20
[0042] prepared according to the reactions: 21
[0043] other jasmonic acid isomers defined according to the
structures: 22
[0044] for example, are also produced in very small quantities when
using the strains of Diplodia gossypina:
[0045] (i) Diplodia gossypina ATCC 10936;
[0046] (ii) Diplodia gossypina ATCC 20575;
[0047] (iii) Diplodia gossypina NRRL 25011; and
[0048] (iv) Diplodia gossypina ATCC 36037.
[0049] The inventive process may be conducted in a batch or
continuous mode of operation. In a batch fermentation, the nutrient
broth, culture and substrate, are combined and fermented until the
jasmonic acid concentration becomes constant. In a continuous
process, the substrate in the nutrient broth may be continuously
recirculated through a fermentation reactor, with the provision
that substrate and product are respectively added and removed from
the recirculating broth.
[0050] In carrying out the present invention, cultivation and
fermentive incubation of the Diplodia gossypina fungus are
accomplished in an aqueous medium in the presence of the usual
nutrient substances (preferably using sodium nitrate, magnesium
sulfate heptahydrate and glucose). A suitable medium thus is one
which contains carbon sources, nitrogen sources, inorganic salts
and growth factors. Additional examples of inorganic salts include
the phosphate salts of magnesium and the phosphate and sulfate
salts of sodium, calcium and potassium. These nutrients may be
supplemented with, for example, one or more vitamins of the "B"
group and one or more trace minerals such as iron, manganese,
cobalt and copper, as desired.
[0051] For the nutrient broth, it is preferred to utilize glucose
(as stated, supra) at a concentration of from about 2 up to about
20 weight percent preferably at about 10 weight percent. It is also
preferred to employ "B" vitamins either as a separate supplement or
in the form of a yeast extract. The kind and amounts of these
additives can be determined by applying the general knowledge in
the art for the cultivation of microorganisms.
[0052] In a typical procedure, one of the Diplodia gossypina
organisms:
[0053] (i) Diplodia gossypina ATCC 10936;
[0054] (ii) Diplodia gossypina ATCC 20575;
[0055] (iii) Diplodia gossypina NRRL 25011; or
[0056] (iv) Diplodia gossypina ATCC 36037
[0057] is first cultivated in inoculum quantifies to produce a
mature culture in nutrient broth. The culture is inoculated into a
fermentor nutrient broth and allowed to establish itself. The
substrate is then added and fermentation is continued until a
steady concentration of jasmonic acid is present.
[0058] The cultivation and fermentative incubation of the Diplodia
gossypina fungus can be carried out as a stationary culture or as a
submerged culture (e.g., shake-flask fermentor) under aerobic
conditions. Cultivation and incubation may proceed as stated,
supra, in a pH range of from about 4.5 up to about 9, preferably at
6. The pH may be regulated by the addition of an inorganic or
organic acid or base such as hydrochloric acid, acetic acid, sodium
hydroxide, calcium carbonate, ammonia, ion-exchange resins or by
the addition of a buffer such as phosphate or phthalate. The
incubation temperature as stated, supra, is suitably maintained
between about 20.degree. C. up to about 35.degree. C., with a range
of from about 25.degree. C. up to about 32.degree. C. being
preferred.
[0059] In accordance with another typical procedure of the present
invention, the process is conveniently carried out by adding the
substrate to the culture medium at the onset of cultivation under
aerobic conditions. Alternatively, the substrate may be added
either alone or in combination with another carbon source, such as
xylose during fermentative incubation or when cultivation is
complete. It is preferable to add the substrate to the culture
medium during the period of from 4 up to 24 hours after the growth
of the culture in the fermentative broth has commenced. Desirable
results can be obtained when the substrate is added continuously
over the entire fermentation after an initial fungal (Diplodia
gossypina) cultivation period of from 3 up to 12 hours. A preferred
feed rate for this continuous addition is from about 0.01 up to I
gram per hour per liter, with a preferred range of from 0.6 up to
0.8 grams per hour per liter. The concentration of the substrate in
the medium may vary depending on the conditions employed. In
practice, the concentration of the substrate in the medium may
conveniently vary from 0.01% up to about 10%/o, with a preferable
concentration of about 1% by weight consistent with the manner in
which it is added to the culture.
[0060] The present invention provides unexpectedly high yields of
jasmonic acid, e.g., from about 0.8 up to about 1.5 grams per
liter.
[0061] Another aspect of our invention is the production of methyl
jasmonate isomers 23
[0062] In this case, the isolation step comprises the steps of:
[0063] (a) extraction of the jasmonic acid from the fermentation
broth with an extraction solvent such as ethyl acetate to form a
jasmonic acid extract;
[0064] (b) concentration of the jasmonic acid extract whereby the
extraction solvent is stripped;
[0065] (c) esterification of the resulting jasmonic acid with
methyl alcohol whereby the methyl jasmonate is formed according to
one of the reactions: 24
[0066] (d) concentration of the resulting methyl jasmonate.
[0067] Additional steps of fractionation of the resulting
concentrate with, for example, silica gel in order to effect isomer
separation and then collecting the 98% "cis" isomer are
preferred.
[0068] The jasmonic acid derivative(s) defined according to the
structure: 25
[0069] (wherein R is methyl or hydrogen) and one or more auxiliary
perfume ingredients including, for example, hydrocarbons, alcohols,
ketones (other than the jasmonic acid derivatives of our
invention), aldehydes, nitriles, esters (other than the jasmonic
acid derivatives of our invention), ethers, synthetic essential
oils and natural essential oils, may be admixed so that the
combined odors of the individual components produce a pleasant and
desired fragrance, particularly and preferably in the floral area
(e.g., jasmine and jasmine/rose aromas). Such perfume compositions
usually contain (a) the main note or the "bouquet" or foundation
stone of the composition; (b) modifiers which round off and
accompany the main note; (c) fixatives which include odorous
substances which lend a particular note to the perfume throughout
all stages of evaporation, and substances which retard evaporation;
and (d) topnotes which are usually low-boiling, fresh-smelling
materials.
[0070] In perfume compositions, it is the individual components
which contribute to their particular olfactory characteristics;
however, the overall sensory effect of the perfume composition will
be at least the sum total of the effects of each of the
ingredients. Thus, one or more of the jasmonic acid derivative(s)
of our invention can be used to alter, modify or enhance the aroma
characteristics of a perfume composition, for example, by utilizing
or moderating the olfactory reaction contributed by another
ingredient in the composition. Thus, a suitable mixture would be
that of the cis isomers of jasmonic acid and methyl jasmonate, to
wit: the mixture of the compounds having the structures: 26
[0071] or the mixture of compounds having the structures: 27
[0072] The amount of jasmonic acid derivative(s) of our invention,
which will be effective in perfume compositions as well as in
perfumed articles and colognes, depends upon many factors including
the other ingredients, their amounts and the side effects which are
desired. It has been found that perfume compositions containing as
little as 0.005% of the jasmonic acid derivative(s) or even less
(e.g., 0.002%) can be used to impart powerful long lasting jasmine,
floral-herbaceous aromas with sweet-herbaceous, green-woody
topnotes to soaps, cosmetics, detergents including anionic,
cationic, nonionic and zwitterionic solid or liquid detergents,
perfumed polymers and other products. The amounts employed can
range up to 70% of the fragrance components and will depend upon
considerations of cost, nature of the end product, the effect
desired on the finished product and the particular fragrance
sought.
[0073] The jasmonic acid derivative(s) of our invention are useful
(taken alone or taken together with other ingredients in perfume
compositions) in detergents, soaps, space odorants and deodorants,
perfines, colognes, toilet waters, bath preparations, hair
preparations such as lacquers, brilliantines, pomades and shampoos;
cosmetic preparations such as creams, deodorants, hand lotions and
sun screens; powders such as talcs, dusting powders, face powders
and the like.
[0074] As little as 0.25% of the jasmonic acid derivative(s) will
suffice to impart an intense and long lasting jasmine,
floral-herbaceous aroma with sweet-herbaceous, green-woody topnotes
to floral perfume formulations. Generally no more than 5% of the
jasmonic acid derivative(s) based on the ultimate end product are
required to be used in the perfume compositions.
[0075] Furthermore, as little as 0.25% of the jasmonic acid
derivative(s) will suffice to impart such aromas to perfumed
articles per se, whether in the presence of other perfume materials
or whether used by themselves. Thus, the range of use of the
jasmonic acid derivative(s) of our invention in perfumed articles,
e.g., perfumed polymers and solid or liquid anionic, cationic,
nonionic or zwitterionic solid or liquid detergents, may vary from
0.25% up to about 5% by weight based on the total weight of the
perfumed article.
[0076] In addition, the perfume composition or fragrance
composition of our invention can contain a vehicle or carrier for
the jasmonic acid derivative(s). The vehicle can be a liquid such
as a nontoxic alcohol, e.g., ethanol; a nontoxic glycol, e.g.,
propylene glycol; or the like. The carrier can also be an absorbent
solid such as a gum (e.g., gum arabic, xanthan gum or guar gum) or
components for encapsulating the composition by means of
coacervation (such as gelatin) or by means of formation of a
polymer around a liquid center (as by using a urea formaldehyde
prepolymer to form a polymeric capsule around a perfume composition
center).
[0077] It will be appreciated from the present disclosure that the
jasmonic acid derivative(s) according to the present invention can
be used to alter, vary, fortify, modify, enhance or otherwise
improve the flavor of a wide variety of materials which are
ingested, consumed or otherwise organoleptically sensed.
[0078] The terms "alter" and "modify" in their various forms will
be understood herein to mean the supplying or imparting of a flavor
character or note to an otherwise bland, relatively tasteless
substance, or augmenting an existing flavor characteristic where
the natural flavor is deficient in some regard or supplementing the
existing flavor impression to modify its organoletpic
character.
[0079] The term "enhance" is intended herein to mean the
intensification (by use of the jasmonic acid derivative of our
invention) of a flavor or aroma note or nuance in a tobacco flavor
or foodstuff or perfume composition or perfumed article without
changing the quality of said note or nuance.
[0080] A "flavoring composition" is taken to mean one which
contributes a part of the overall flavor impression by
supplementing or fortifying a natural or artificial flavor in a
material, or one which supplies substantially all the flavor and/or
aroma character to a consumable article.
[0081] The term "foodstuff" as used herein includes both solid and
liquid ingestible materials for man or animals, which materials
usually do, but need not, have nutritional value. Thus, foodstuffs
include meats, gravies, soups, convenience foods, malt, alcoholic
and other beverages, milk and dairy products, seafoods, including
fish, crustaceans, mollusks and the like, candies, vegetables,
cereals, soft drinks, snacks, dog and cat foods, other veterinary
products and the like. The jasmonic acid derivative(s) of our
invention are also useful in tobacco flavorants and tobacco
enhancers.
[0082] The term "tobacco" will be understood herein to mean natural
products such as, for example, Burley, Turkish tobacco, Maryland
tobacco, flue-cured tobacco and the like, including tobacco-like or
tobacco-based products such as reconstituted or homogenized leaf
and the like, as well as tobacco substitutes intended to replace
natural tobacco such as lettuce and cabbage leaves and the like.
The tobaccos and tobacco products in which the jasmonic acid
derivative(s) of our invention are useful include those designed or
used for smoking such as in cigarettes, cigar and pipe tobacco, as
well as products such as snuff, chewing tobacco and the like.
[0083] When the jasmonic acid derivative(s) of this invention are
used in a flavoring composition, they can be combined with
conventional flavoring materials or adjuvants. Such co-ingredients
or flavor adjuvants are well known in the art for such use and have
been extensively described in the literature. Requirements of such
adjuvant materials are: (1) that they be non-reactive with the
jasmonic acid derivative(s) of our invention; (2) that they be
organoleptically compatible with the jasmonic acid derivative(s) of
our invention whereby the flavor of the ultimate consumable
material to which the jasmonic acid derivative(s) are added is not
detrimentally affected by the use of the adjuvant; and (3) that
they be ingestibly acceptable and thus nontoxic or otherwise
non-deleterious. Apart from these requirements, conventional
materials can be used and broadly include other flavor materials,
vehicles, stabilizers, thickeners, surface active agents,
conditioners and flavor intensifiers.
[0084] Such conventional flavoring materials include saturated
fatty acids, unsaturated fatty acids and amino acids; alcohols
including primary and secondary alcohols, esters, carbonyl
compounds including ketones (other than the jasmonic acid
derivatives of our invention) and aldehydes; lactones; other cyclic
organic materials including benzene derivatives, alicyclic
compounds, heterocyclics such as furans, pyridines, pyrazines and
the like; sulfur-containing compounds including thiols, sulfides,
disulfides and the like; proteins; lipids, carbohydrates; so-called
flavor potentiators such as monosodium glutamate; magnesium
glutamate, calcium glutamate, guanylates and inosinates; natural
flavoring materials such as cocoa, vanilla and caramel; essential
oils and extracts such as anise oil, clove oil and the like and
artificial flavoring materials such as vanillin, ethyl vanillin and
the like.
[0085] Specific preferred flavor adjuvants are as follows:
[0086] anise oil;
[0087] ethyl-2-methyl butyrate;
[0088] vanillin;
[0089] cis-3-heptenol;
[0090] cis-3-hexenol;
[0091] trans-2-heptenal;
[0092] butyl valerate;
[0093] 2,3-diethyl pyrazine;
[0094] methyl cyclopentenolone;
[0095] benzaldehyde;
[0096] valerian oil;
[0097] 3,4-dimethoxyphenol;
[0098] amyl acetate;
[0099] amyl cinnamate;
[0100] .gamma.-butyryl lactone;
[0101] furfural;
[0102] trimethyl pyrazine;
[0103] phenyl acetic acid;
[0104] isovaleraldehyde;
[0105] ethyl maltol;
[0106] ethyl vanilin;
[0107] ethyl valerate;
[0108] ethyl butyrate;
[0109] cocoa extract;
[0110] coffee extract;
[0111] peppermint oil;
[0112] spearmint oil;
[0113] clove oil;
[0114] anethol;
[0115] cardamom oil;
[0116] wintergreen oil;
[0117] cinnamic aldehyde;
[0118] ethyl-2-methyl valerate;
[0119] .gamma.-hexenyl lactone;
[0120] 2,4-decadienal;
[0121] 2,4-heptadienal;
[0122] methyl thiazole alcohol (4-methyl-5-.beta.-hydroxyehtyl
thiazole);
[0123] 2-methyl butanethiol;
[0124] 4-mercapto-2-butanone;
[0125] 3-mercapto-2-pentanone;
[0126] 1-mercapto-2-propane;
[0127] benzaldehyde;
[0128] furfural;
[0129] furfuryl alcohol;
[0130] 2-mercapto propionic acid;
[0131] alkyl pyrazine;
[0132] methyl pyrazine;
[0133] 2-ethyl-3-methyl pyrazine;
[0134] tetramethyl pyrazine;
[0135] polysulfides;
[0136] dipropyl disulfide;
[0137] methyl benzyl disulfide;
[0138] alkyl thiophene;
[0139] 2,3-dimethyl thiophene;
[0140] 5-methyl furfural;
[0141] acetyl furfuran;
[0142] 2,4-decadienal;
[0143] guiacol;
[0144] phenyl acetaldehyde;
[0145] .beta.-decalactone;
[0146] d-limonene;
[0147] acetoin;
[0148] amyl acetate;
[0149] maltol;
[0150] ethyl butyrate;
[0151] levulinic acid;
[0152] piperonal;
[0153] ethyl acetate;
[0154] n-octanal;
[0155] n-pentanal;
[0156] n-hexanal;
[0157] diacetyl;
[0158] monosodium gulatamate;
[0159] monopotassium glutamate;
[0160] sulfur-containing amino acids, e.g., cysteine;
[0161] hydrolyzed vegetable protein;
[0162] 2-methylfuran-3-thiol;
[0163] 2-methyldihydrofuran-3-thiol;
[0164] 2,5-dimethylfuran-3-thiol;
[0165] hydrolyzed fish protein;
[0166] tetrametiyl pyrazine;
[0167] propylpropenyl disulfide;
[0168] propylpropenyl trisulfide;
[0169] diallyl disulfide;
[0170] diallyl trisulfide;
[0171] dipropenyl disulfide;
[0172] dipropenyl trisulfide;
[0173] 4-methyl-2-[(methylthio)-ethyl]-1,3-dithiolane;
[0174] 4,5-dimethyl-2-(methylthiomethyl)-1,3-dithiolne; and
[0175] 4-methyl-2-(methylthiomethyl)-1,3-dithiolane.
[0176] The jasmonic acid derivative(s) of our invention or
compositions incorporating them, as mentioned above, can be
combined with one or more vehicles or carriers for adding them to
the particular product. Vehicles can be edible or otherwise
suitable materials such as ethyl alcohol, propylene glycol, water
and the like, as described supra. Carriers include materials such
as gum arabic, carrageenan, xanthan gum, guar gum and the like.
[0177] The jasmonic acid derivative(s) prepared according to our
invention can be incorporated with the carriers by conventional
means such as spray-drying, drum-drying and the like. Such carriers
can also include materials for coacervating the jasmonic acid
derivative(s) of our invention to provide encapsulated products, as
set forth supra. When the carrier is an emulsion, the flavoring
composition can also contain emulsifiers such as mono- and
diglycerides or fatty acids and the like. With these carriers or
vehicles, the desired physical form of the compositions can be
prepared.
[0178] The quantity of jasmonic acid derivative(s) utilized should
be sufficient to impart the desired flavor characteristic to the
product, but on the other hand, the use of an excessive amount of
the jasmonic acid derivative(s) is not only wasteful and
uneconomical, but in some instances, too large a quantity may
unbalance the flavor or other organoleptic properties of the
product consumed. The quantity used will vary depending upon the
ultimate foodstuff; the amount and type of flavor initially present
in the foodstuff; the further process or treatment steps to which
the foodstuff will be subjected; regional and other preference
factors; the type of storage, if any, to which the product will be
subjected; and the preconsumption treatment such as baking, frying
and so on, given to the product by the ultimate consumer.
Accordingly, the terminology "effective amount" and "sufficient
amount" is understood in the context of the present invention to be
quantitatively adequate to alter the flavor of the foodstuff.
[0179] It is accordingly preferred that the ultimate composition
contain from about 0.1 parts per million (ppm) up to about 500 ppm
of the jasmonic acid derivative(s).
[0180] The jasmonic acid derivative(s) of our invention when
utilized in flavoring compositions can be varied over a wide range
depending upon the particular flavor nuances desired to be added to
the foodstuff. Thus, the amounts of jasmonic acid derivative(s) of
our invention may be contained in flavoring materials from about 1
ppm up to about 50% by weight of the flavoring composition. Indeed,
the compounds having the structures: 28
[0181] may be utilized in raspberry flavors (e.g., for use in
raspberry-flavored yogurt, for example) at levels of between about
1% and about 50%. Such materials also have utility in flavorings
for salad dressings, particularly in such cuisines as "Thai"
cuisine.
[0182] According to another aspect of our invention, an
organoleptically improved smoking tobacco product and additives
therefor as well as methods of making the same which overcome
specific problems heretofore encountered in which specific Turkish,
oriental-like aromas prior to smoking and improved Turkish,
oriental aromas on smoking in the mainstream and the side stream
are created or enhanced or modified or augmented and may be readily
controlled and maintained at the desired uniform level regardless
of variations in the tobacco components of the blend. In
particular, low grade Virginia-type tobaccos may be upgraded using
the jasmonic acid derivative(s) of our invention.
[0183] This invention further provides improved tobacco additives
and methods whereby various desirable natural aromatic Turkish
tobacco flavoring characteristics with oriental notes may be
imparted to smoking tobacco products and may be readily varied and
controlled to produce the desired uniform flavoring
characteristics.
[0184] In carrying out this aspect of our invention, we add to
smoking tobacco materials or a suitable substitute therefor (e.g.,
dried lettuce leaves) an aroma and flavor additive containing as an
active ingredient one or more of the jasmonic acid derivative(s) of
our invention.
[0185] In addition to the jasmonic acid derivative(s) of our
invention, other flavoring and aroma additives may be added to the
smoking tobacco material or substitute therefor, either separately
or in admixture with the jasmonic acid derivative(s) of our
invention as follows:
I. Synthetic Materials
[0186] .beta.-Ethyl-cinnamaldehyde;
[0187] Eugenol;
[0188] Dipentene;
[0189] .beta.-Damascenone;
[0190] Maltol;
[0191] Ethyl maltol;
[0192] Delta-Undecalactone;
[0193] Delta-Decalactone;
[0194] Benzaldehyde;
[0195] Amyl acetate;
[0196] Ethyl butyrate;
[0197] Ethyl valerate;
[0198] Ethyl acetate;
[0199] 2-Hexenol-1;
[0200] 2-Methyl-5-isopropyl-1,3-nonadiene-8-one;
[0201] 2,6-Dimethyl-1,6-undecadiene-10-one;
[0202] 2-ethyl-5-isopropyl acetophenone;
[0203]
2-Hydroxy-2,5,5,8a-tetramethyl-1-(2-hydroxyethyl)-decahydrmonaphtha-
lene;
[0204] Dodcahydro-3a,6,6,9a-tetramethyl naphthol(2, 1-b) furan;
[0205] 4-Hydroxy hexanoic acid, .gamma.-lactone; and
[0206] Polyisoprenoid hydrocarbons defined in Example V of U.S.
Pat. No. 3,589,372 issued on Jun. 29, 1971.
II. Natural Oils
[0207] Celery seed oil;
[0208] Coffee extract;
[0209] Bergamot oil;
[0210] Cocoa extract;
[0211] Nutmeg oil; and
[0212] Origanum oil.
[0213] An aroma and flavoring concentrate containing one or more of
the jasmonic acid derivative(s) of our invention and, if desired,
one or more of the above-indicated additional flavoring additives
may be added to the smoking tobacco material, to the filter or to
the leaf or paper wrapper. The smoking tobacco material may be
shredded, cured, cased and blended tobacco material or
reconstituted tobacco material or tobacco substitutes (e.g.,
lettuce leaves) or mixtures thereof. The proportions of flavoring
additives may be varied in accordance with taste, but insofar as
enhancement or the imparting of oriental and/or Turkish tobacco
notes, we have found that satisfactory results are obtained if the
proportion by weight of the sum total of jasmonic acid
derivative(s) to smoking tobacco material is between 50 ppm and
1,500 ppm (0.005%-0.15%) of the active ingredients to the smoking
tobacco material. We have further found that satisfactory results
are obtained if the proportion by weight of the sum total of
jasmonic acid derivative(s) used to flavoring material is between
500 and 15,000 ppm (0.05%-1.5%).
[0214] Any convenient method for incorporating the jasmonic acid
derivative(s) into the tobacco product may be employed. Thus, the
jasmonic acid derivative(s) taken alone or along with other
flavoring additives may be dissolved in a suitable solvent such as
ethanol, diethyl ether and/or other organic solvents, and the
resulting solution may either be spread onto the cured, cased and
blended tobacco material or the tobacco material may be dipped into
such solution. Under certain circumstances, a solution of the
jasmonic acid derivative(s) taken alone or taken further together
with flavoring additives as set forth above may be applied by means
of a suitable applicator such as a brush or roller on the paper or
leaf wrapper for the smoking product, or it may be applied to the
filter by either spraying or dipping or coating.
[0215] Furthermore, it will be apparent that only a portion of the
tobacco or substitute therefor need be treated, and the
thus-treated tobacco may be blended with other tobaccos before the
ultimate tobacco product is formed. In such cases, the tobacco
treated may have the jasmonic acid derivative(s) in excess of the
amounts or concentrations above indicated so that when blended with
other tobaccos, the final product will have the percentage within
the indicated range.
[0216] In accordance with one specific example of our invention, an
aged, cured and shredded domestic Virginia tobacco is sprayed with
a 20% alcohol solution of the compound having the structure: 29
[0217] optical rotation (.alpha..sub.D.sup.20) equal +58.degree. on
a dry basis. Thereafter, the alcohol is removed by evaporation and
the tobacco is manufactured into cigarettes by the usual
techniques. The cigarette, when treated as indicated, has a desired
and pleasing aroma which is detectable in the mainstream and the
side stream when the cigarette is smoked. The aroma is described as
being sweeter with pronounced Turkish/oriental characteristics and
with improved body and enhanced tobacco character with subsidiary
"floral," "jasmonic" nuances in the mainstream and the side stream.
In addition, interesting amber nuances are inparted.
[0218] While our invention is particularly useful in the
manufacture of smoking tobacco such as cigarette tobacco, cigar
tobacco and pipe tobacco, other tobacco products formed from
sheeted tobacco dust or fines may also be used. Likewise, the
jasmonic acid derivative(s) of our invention can be incorporated
with materials such as filter tip materials, seam paste, packaging
materials and the like, which are used along with tobacco to form a
product adapted for smoking. Furthermore, the jasmonic acid
derivative(s) can be added to certain tobacco substitutes of
natural or synthetic origin (e.g., dried lettuce leaves) and,
accordingly, by the term "tobacco" as used throughout this
specification, is meant any composition intended for human
consumption by smoking or otherwise when composed of tobacco plant
parts or substitute material or both. Thus, chewing tobacco is also
included in the foregoing meaning of "tobacco."
[0219] During the fermentation reaction of our invention, two
byproducts are also formed, to wit, the compound having the
structure: 30
[0220] and the compound having the structure: 31
[0221] the compound having the structure: 32
[0222] formed via the reaction: 33
[0223] It will be understood that each of the compounds having the
structures: 34
[0224] may be retained with the jasmonic acid derivative(s) of our
composition for their flavor or fragrance utilities. Thus, in
addition to the additives as set forth, supra, the compounds having
the structures: 35
[0225] may be "retained" and not "separated" in the isolation of
the jasmonic acid derivative(s) prior to the jasmonic acid
derivative(s) being utilized in flavor or fragrance
formulations.
[0226] More specifically, our invention also contemplates mixtures
of derivatives having the structures: 36
[0227] (wherein R is methyl or hydrogen) taken further together
with the compounds having the structures: 37
BRIEF DESCRIPTION OF THE DRAWINGS
[0228] FIG. 1 is the HPLC profile for the reaction product of
Example XIII (conditions: 250 mm.times.4.6 mm ZORBAX.RTM. SB-C18
column).
[0229] FIG. 2 is the HPLC profile for the reaction product of
Example XIV (conditions: 250 mm.times.4.6 mm ZORBAX.RTM. SB-C18
column).
[0230] FIG. 3 is the HPLC profile for the reaction products of
Example XV having the structures: 38
[0231] FIG. 4 is the NMR spectrum for the product of Example XV for
the compound having the structure: 39
[0232] (optical rotation (.alpha..sub.D.sup.20)=+58).
[0233] FIG. 4A is an enlargement of section "A" of the NMR spectrum
of FIG. 4.
[0234] FIG. 4B is an enlargement of section "B" of the NMR spectrum
of FIG. 4.
[0235] FIG. 4C is an enlargement of section "C" of the NMR spectrum
of FIG. 4.
[0236] FIG. 5A is a GC mass spectrum for tie compound having the
structure: 40
[0237] (.alpha..sub.D.sup.20) prepared according to Example XV.
[0238] FIG. 5B is the GC mass spectrum for the product produced
according to Example XV having the structure: 41
[0239] together with small amounts of the compound defined
according to tie structure: 42
[0240] with tie optical rotation for tie compound having tie
structure: 43
[0241] being (.alpha..sub.D.sup.20)+580.
[0242] FIG. 6 is the mass spectrum for the compound having the
structure: 44
[0243] prepared according to Example XV.
[0244] FIG. 7 is the NMR spectrum for the compound having the
structure: 45
[0245] used in the process of Example XIII.
[0246] FIG. 8 represents a cutaway side elevation view of apparatus
used in forming perfumed polymers which contain embedded therein at
least one of the jasmonic acid derivative-containing compositions
of our invention.
[0247] FIG. 9 is a front view of the apparatus of FIG. 8 looking in
the direction of the arrows.
[0248] FIG. 10A is a schematic block flow diagram showing process
steps for preparing pure jasmonic acid defined according to the
structures: 46
[0249] FIG. 10B is a schematic block flow diagram showing the
process steps and apparatus for preparing the methyl jasmonate of
our invention from the jasmonic acid of our invention according to
the reactions: 47
[0250] FIG. 11 is the NMR spectrum for the pure optical isomer
(.alpha..sub.D.sup.20) of the compound having the structure: 48
[0251] prepared according to Example XV having an optical rotation
of +58.degree..
[0252] FIG. 11A is an enlargement of section "A" of the NMR
spectrum of FIG. 11.
[0253] FIG. 11B is an enlargement of section "B" of the NMR spectum
of FIG. 11
[0254] FIG. 11C is an enlargement of section "C" of the NMR
spectrum of FIG. 11.
[0255] FIG. 12 is the mass spectrum for the reaction product of
Example XV containing the compounds having the structures: 49
[0256] as well as the compounds having the structures: 50
[0257] and the compound having the structure: 51
[0258] FIG. 13A is the HPLC profile for the optical isomer having
the structure: 52
[0259] prepared according to Example XV (optical rotation
(.alpha..sub.D.sup.20)=+58.degree.) (conditions: OV1 column).
[0260] FIG. 13B is the HPLC profile for the optical isomer
(.alpha..sub.D.sup.20=+58.degree.) having the structure: 53
[0261] prepared according to Example XV (conditions: Carbowax
column).
[0262] FIG. 14 is the HPLC profile (chiral column) for the optical
isomer (.alpha..sub.D.sup.20=+58.degree.) of the compound having
the structure: 54
[0263] produced according to Example XV.
[0264] FIG. 15 is the NMR spectrum for the compound having the
structure: 55
[0265] (optical isomer: (.alpha..sub.D.sup.20=+58.degree.) prepared
according to Example VI.
[0266] FIG. 15A is an enlargement of section "A" of the NMR
spectrum of FIG. 15.
DETAILED DESCRIPTION OF THE DRAWINGS
[0267] Referring to FIG. 1, the peak indicated by reference numeral
10 is the peak for the jasmonic acid isomer defined according to
the structure: 56
[0268] Referring to FIG. 2, the peak indicated by reference numeral
20 is the peak for the jasmonic acid defined according to the
structure: 57
[0269] Referring to FIG. 2, the peak indicated by reference numeral
41 is the peak for the isomer having the structure: 58
[0270] The peak indicated by reference numeral 31 is for the isomer
having the structure: 59
[0271] The curve for the material containing the isomer having the
structure: 60
[0272] is indicated by reference numeral 30. The curve for the
material containing the isomer having the structure: 61
[0273] is indicated by reference numeral 40.
[0274] Referring to FIG. 5B, the peak indicated by reference
numeral 50 is the peak for the isomer having the structure: 62
[0275] The peaks indicated by reference numerals 51 and 52 are for
the isomer having the structure: 63
[0276] Referring to FIGS. 8 and 9, there is provided a process for
forming scented polymer elements (wherein the polymer may be a
thermoplastic polymer such as low density polyethylene or
polypropylene or copolymers of ethylene and vinyl acetate or
mixtures of polymers and copolymers such as copolymers of ethylene
and vinyl acetate and polyethylene) such as pellets useful in the
formation of plastic particles useful in fabricating certain
articles which may be perfumed. This process comprises heating the
polymer or mixture of polymers to the melting point of said polymer
or mixture of polymers, e.g., 250.degree. C. in the case of low
density polyethylene. The lower most portion of the container is
maintained at a slightly lower temperature and the material in the
container is taken off at such location for delivery through the
conduit. Thus referring to FIGS. 8 and 9, in particular, the
apparatus used in producing such elements comprises a device for
forming the polymer containing perfume, e.g., polyethylene or
polyethylene-polyvinyl acetate of mixtures of same or
polypropylene, which comprises a vat or container 1212 into which
the polymer taken alone or in admixture with other polymers in the
perfuming substance which is at least one of the jasmonic acid
derivative(s) of our invention or mixtures of jasmonic acid
derivatives and other compatible perfume components is placed. The
container is closed by means of an airtight lid 1228 and clamped to
the container by bolts 1265. A stirrer 1273 traverses the lid or
cover 1228 in an airtight manner and is rotatable in a suitable
manner. A surrounding cylinder 1212A having heating coils which are
supplied with electric current through cable 1214 from a rheostat
or control 1216 is operated to maintain the temperature inside the
container 1212 such that the polymer in the container will be
maintained in the molten or liquid state. It has been found
advantageous to employ polymers at such a temperature that the
viscosity will be in the range of 90-100 Saybolt seconds. The
heater 1218 is operated to maintain the upper portion of the
container 1212 within a temperature range of, for example,
220-270.degree. C. in the case of low density polyethylene. The
bottom portion of the container 1212 is heated by means of heating
coils 1212A regulated through the control 1220 connected thereto
through a connecting wire 1222 to maintain the lower portion of the
container 1212 within a temperature range of 220-270.degree. C.
[0277] Thus, the polymer or mixture of polymers added to the
container 1212 is heated from 10-12 hours, whereafter the perfume
composition or perfume material which contains one or more of the
jasmonic acid derivatives of our invention is quickly added to the
melt. Generally, about 10-45% by weight of the resulting mixture of
the perfumery substance is added to the polymer.
[0278] After the perfume material is added to the container 1212,
the mixture is stirred for a few minutes, for example, 5-15
minutes, and maintained within the temperature ranges indicated
previously by the heating coil 1212A. The controls 1216 and 1220
are connected through cables 1224 and 1226 to a suitable supply of
electric current for supplying the power for heating purposes.
[0279] Thereafter, the valve "V" is opened, permitting the mass to
flow outwardly through conduit 1232 having a multiplicity of
orifices 1234 adjacent to the lower side thereof. The outer end of
the conduit 1232 is closed so that the liquid polymer in intimate
admixture with one or more of the jasmonic acid derivatives of our
invention or mixture of perfume substance and one or more of the
jasmonic acid derivatives of our invention, will continuously drop
through the orifices 1234 downwardly from the conduit 1232. During
this time, the temperature of the polymer intimately admixed with
the perfumery substance in the container 1212 is accurately
controlled so that a temperature in the range of from about
240-250.degree. C. (for example, in the case of low density
polyethylene) will exist in the conduit 1232. The regulation of the
temperature through the controls 1216 and 1220 is essential in
order to insure temperature balance to provide for the continuous
dropping or dripping of molten polymer intimately admixed with the
perfiume substance which is all or which contains one or more of
the jasmonic acid derivatives of our invention, through the
orifices 1234 at a rate which will insure the formation of droplets
1236 which will fall downwardly onto a moving conveyor belt 1238
caused to run between conveyor wheels 1240 and 1242 beneath the
conduit 1232.
[0280] When the droplets 1236 fall onto the conveyor 1238, they
form pellets 1244 which harden almost instantaneously and fall off
the end of the conveyor 1238 into a container 1245 which is
advantageously filled with water or some other suitable cooling
liquid to insure the rapid cooling of each of the pellets 1244. The
pellets 1244 are then collected from the container 1245 and
utilized for the formation of other functional products, e.g.,
garbage bags and the like.
[0281] Belt 1238 is continuously moistened with sponge-like
material 1256 supplied with water 1254 from container 1250 having
sidewall 1248 so that the belt is continuously cooled.
[0282] Referring to FIG. 10A, carbon source (e.g., glucose) from
location 102 is admixed with nutrient (containing, for example,
magnesium sulfate heptahydrate and sodium nitrate) from location
101 and culture (containing at least one strain of Diplodia
gossypina) from location 100 are admixed and placed in fermenter
103. The fermenter is run, for example, for a period of 10 days,
and the resulting product is then passed through line 106 past
valve 107 into mixing vessel 105 and combined with extraction
solvent (e.g., ethyl acetate) from location 104. The resulting
mixture is then separated and the solvent/jasmonic acid material is
passed through line 110 past valve 111 into distillation column 112
where overhead solvent is distilled through line 113 and recovered
at location 114, and crude jasmonic acid (bottoms) is passed into
vessel 119 where it is admixed with aqueous sodium carbonate
solution from location 116 which is passed through line 117 past
valve 118. The resulting sodium carbonate solution/jasmonic acid
material is then passed through line 121 past valve 122 and mixed
with extraction solvent from location 120 which is passed through
line 123 past valve 124 and admixed with such extraction solvent in
vessel 125. The organic phase is then passed to location 127
through line 126, and the aqueous phase is passed through line 128
into vessel 131 where it is admixed with phosphoric acid from
location 129 (pH adjustment). The resulting product is then further
mixed with extraction solvent from location 132 which is passed
through line 133 past valve 134 and admixed in vessel 136. The
resulting organic phase is separated from the aqueous phase and
passed through line 137 past valve 138 into distillation column
139, where the recovered solvent (overhead) is passed through line
140 into solvent recovery system 141, and the resulting pure
jasmonic acid (bottoms) is passed through line 142 into location
143 (structure: 64
[0283] Referring to FIG. 10B, jasmonic acid having the structure:
65
[0284] (from the apparatus of FIG. 10A) at location 143 is passed
through line 145 past valve 146 into autoclave 149, where it is
admixed with methanol from location 144 which is passed through
line 147 past valve 148 into autoclave 149. Autoclave 149 is heated
under pressure using heating element 150 causing the reactions:
66
[0285] to take place thereat. The resulting products (crude) having
the structures: 67
[0286] are then passed through line 151 past valve 152 into
distillation column 153, where the excess methanol is recovered
overhead via line 154 into recovery system 155, and the resulting
crude methyl jasmonate products are admixed with extraction solvent
from location 156, passed through line 157 past valve 160 into
vessel 159. The extraction solvent/crude methyl jasmonate is then
mixed and passed through line 161 past valve 162 into distillation
column 163, where solvent is recovered overhead via line 164 into
solvent recovery system 165, and purified methyl jasmonate
(bottoms) is passed into vessel 167, the purified methyl jasmonate
having the structures: 68
[0287] Referring to FIG. 12, the GC mass spectrum for the crude
reaction product of Example XV contains the following peaks:
[0288] (a) the peak indicated by reference numeral 120 is the peak
for the compound having the structure: 69
[0289] (b) the peak indicated by reference 121 is the peak for the
compound having the structure: 70
[0290] (c) the peak indicated by reference numeral 121 also
contains a side peak for the compound having the structure: 71
[0291] (d) the peaks indicated by reference numerals 122 and 123
are for compounds having the structures: 72
[0292] (e) the peak indicated by reference numeral 124 is for the
compound having the structure: 73
[0293] (f) the peaks indicated by reference numerals 125A and 125B
are for compounds defined according to the structure: 74
[0294] (g) the peak indicated by reference numeral 126 is for tie
compound having the structure: 75
[0295] The following examples are given to illustrate embodiments
of the invention as it is preferred to practice it. It will be
understood that these examples are illustrative, and the invention
is not to be considered as restrictive thereto except as indicated
in the appended claims.
[0296] All parts, proportions, percentages and ratios hereinafter
referred to are by weight unless otherwise indicated.
EXAMPLE I
Jasmonic Acid Screening Procedure
[0297]
1 Medium 5059H KH.sub.2PO.sub.4 1.0 gm MgSO.sub.4.cndot.7H.sub.2O
0.5 gm Yeast extract 1.0 gm Soy peptone 5.0 gm Trace minerals 10.0
ml Dextrose 30.0 gm Deionized water 1.0 L pH adjusted to 5.5 before
sterilization
[0298]
2 Trace Minerals Solution 5059H FeSO.sub.4.7H.sub.2O 10.0 mg
ZnSO.sub.4.7H.sub.2O 8.8 mg CuSO.sub.4.5H.sub.2O 15.0 mg
MnSO.sub.4.H.sub.2O 7.6 mg
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.sub.2O 10.0 mg Deionized water
1.0 L
Procedure
[0299] 500 M1 flasks containing 100 ml of broth were inoculated
from slant stock cultures (PDA) and incubated at 25.degree. C.
stationary or at 150 rpm. Flask cultures were periodically analyzed
by acidifying a 1 ml sample, extracting with 1 ml of ethyl acetate,
centrifuging, concentrating and spotting on TLC plates.
Alternately, a whole flask culture was extracted, the crude extract
used to prepare the methyl ester and followed by GC analysis.
3 TLC Solvents Ethyl acetate 50% Hexanes 50% Acetic acid 0.5%
Screening Results
[0300]
4 TABLE I Jasmonic Acid (mg/L) Incubation Incubation Station- Time
Time Microorganisms ATCC ary (Days) Shake (Days) Diplodia gossypina
10936 210 7 1.1 7 Diplodia gossypina 16391 Neg 7 5.6 7 Diplodia
gossypina 20575 Neg 7 Neg 7 Diplodia gossypina 20576 Neg 7 Neg 7
Diplodia gossypina 22644 Neg 7 Neg 7 Diplodia gossypina 26123 23 7
Neg 7
[0301]
5 TABLE II Jasmonic Acid (TLC) Incubation Incubation Microorganisms
Stationary Time (Days) Shake Time (Days) Diplodia gossypina ATCC
28570 Neg 9 Diplodia gossypina ATCC 34643 + 9 Diplodia gossypina
ATCC 36037 + 9 Diplodia gossypina ATCC 76087 + 9 Lasiodiplodia
theobromae IFO 31643 + 5 Botryosphaeria rhodina CBS 110.11 Neg 7
Botryosphaeria rhodina CBS 124.13 Neg 7 Botryosphaeria rhodina CBS
174.26 Neg 7 Botryosphaeria rhodina CBS 175.26 Neg 7 Botryosphaeria
rhodina CBS 176.26 Neg 7 Botryosphaeria rhodina CBS 190.73 Neg 10
Botryosphaeria rhodina CBS 230.30 Neg 7 Botryosphaeria rhodina CBS
287.47 + 7 Botryosphaeria rhodina CBS 301.36 Neg 7 Botryosphaeria
rhodina CBS 304.79 + 5 Botryosphaeria rhodina CBS 306.58 Neg 5
Botryosphaeria rhodina CBS 356.59 Neg 5 Botryosphaeria rhodina CBS
374.54 Neg 10 Botryosphaeria rhodina CBS 456.78 Neg 10
Botryosphaeria rhodina CBS 494.78 Neg 10 Botryosphaeria rhodina CBS
495.78 Neg 10 Medium 5074H NaNO.sub.3 2.0 gm KH.sub.2PO.sub.4O 1.0
gm MgSO.sub.4.7H.sub.2O 0.5 gm KCl-- 0.5 gm FeSO.sub.4.7H.sub.2O
10.0 mg Yeast extract 1.0 gm Trace minerals 1.0 ml Dextrose 50.0 gm
Deionized water 1.0 L pH adjusted to 5.5 before sterilization Trace
Minerals Solution 5059H ZnSO.sub.4.7H.sub.2O 1.0 gm
CuSO.sub.4.5H.sub.2O 0.15 gm MnSO.sub.4.H.sub.2O 0.1 gm
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.su- b.2O 0.1 gm Deionized water
1.0 L
Screening Results
[0302]
6 TABLE III Jasmonic Acid (TLC) Incubation Incubation
Microorganisms Stationary Time (Days) Shake Time (Days)
Botryodiplodia theobromae FPRL S-22L Neg 7 Neg 7 Botryosphaeria
rhodina CBS 176.26 + 7 + 7 Botryosphaeria rhodina CBS 205.75 + 7 -
7 Botryosphaeria rhodina CBS 287.47 + 7 + 7 Bottyosphaeria rhodina
CBS 304.79 + 7 + 7
[0303]
7TABLE IV NOTE: Same medium as 5074H, except pH adjusted to 9.0
before sterilization. A 72-96 hour shake flask inoculum (100 ml/500
ml flask; 150 rpm; 30.degree. C.) was chopped in a sterile blender
jar and 4 ml was used to inoculate a new flask, including the
addition of 10 ml sterile 50% dextrose. Jasmonic Acid (TIC)
Incubation Incubation Microorganisms Stationary Time (Days) Shake
Time (Days) Diplodia gossypina ATCC 20575 + 6 Diplodia gossypina
ATCC 26123 + 7 Diplodia gossypina ATCC 36037 + 6 Diplodia gossypina
ATCC 64959 + 6 Botryosphaeria rhodina CBS 287.47 + 7 Botryosphaeria
rhodina CBS 304.73 + 6 Lasiodiplodia theobromae IFO 6469 + 7
EXAMPLE II
Screening of Microorganisms in Submerged Culture for the Production
of Jasmonic Acid
[0304]
8 Medium: Buffered Minimal Salts Medium (bMS) NaNO.sub.3 2.0 gm/L
KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5 gm/L
MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L FeSO.sub.4-7H.sub.2O 1.0
mg/L TASTONE .RTM. 900 0.5 gm/L
[0305] After sterilization of bMS medium at 120.degree. C. and 15
psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0306] Inocula for all microorganisms were prepared from frozen
cultures. 2 M1 of inoculum were added to 100 ml of sterile bMS
medium and incubated at 28.degree. C. and 200 rpm. The production
of jasmonic acid in the cultures was monitored by TLC and HPLC
analysis.
9 TLC Solvents Hexane 10% Ethyl acetate 90% Acetic acid 0.5%
[0307]
10 Microorganisms Jasmonic Acid Production (9 Days) Diplodia
gossypina ATCC 10936 648 ppm Diplodia gossypina ATCC 16391 --
Diplodia gossypina ATCC 20575 1263 ppm Diplodia gossyprna ATCC
22644 -- Diplodia gossypina ATCC 26123 -- Diplodia gossypina ATCC
34643 -- Diplodia gossypina ATCC 36037 -- Diplodia gossypina ATCC
64959 -- Diplodia gossypina ATCC 20571 -- Diplodia gossypina NRRL
13607 -- Diplodia gossypina NRRL 25011 75 ppm Diplodia gossypina
ATCC 34643 503 ppm Botryodiplodia theobromae D7/2 337 ppm (German
Patent Stain)
EXAMPLE III
Screening of Microorganisms for the Production of Jasmonic Acid in
Submerged Culture Under Reduced Nitrogen Levels
[0308]
11 Medium: Buffered Minimal Salts Medium (bMS) NaNO.sub.3 0.5 gm/L
KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5 gm/L
MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L FeSO.sub.4-7H.sub.2O 1.0
mg/L TASTONE .RTM. 900 0.5 gm/L
[0309] Following sterilization of bMS medium at 120.degree. C. and
15 psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0310] Inocula for all microorganisms were prepared from frozen
cultures. 2 M1 of inoculum were added to 100 ml of sterile bMS
medium and incubated at 28.degree. C. and 200 rpm. The production
of jasmonic acid in the cultures was monitored by TLC and HPLC
analysis.
12 TLC Solvents Hexane 10% Ethyl acetate 90% Acetic acid 0.5%
[0311]
13 Jasmonic Acid Production Microorganisms (7 Days) Botryodiplodia
rhodina CBS 110.1 - Botryodiplodia rhodina CBS 124.13.1 -
Botryodiplodia rhodina CBS 175.26 - Botryodiplodia rhodina CBS
176.25 44 ppm Botryodiplodia rhodina CBS 190.73 + Botryodiplodia
rhodina CBS 287.47 + Botryodiplodia rhodina CBS 289.56 -
Botryodiplodia rhodina CBS 304.79 + Botryodiplodia rhodina CBS
306.58 + Botryodiplodia rhodina CBS 356.59 - Botryodiplodia rhodina
CBS 374.54 - Botryodiplodia rhodina CBS 447.62 + Botryodiplodia
rhodina CBS 456.78 - Botryodiplodia rhodina CBS 494.78 -
Botryodiplodia rhodina CBS 495.78 - Botryodiplodia rhodina CBS
559.7 + Diplodia gossypina ATCC 36037 417 ppm Diplodia gossypina
ATCC 58760 95 ppm Diplodia gossypina ATCC 76087 +
EXAMPLE IV
Production of Jasmonic Acid in Stationary Fernbach Flask
Culture
[0312] Reactions:
14 76 77 and 78 79 Medium Medium 5074H KH.sub.2PO.sub.4 1.0 gm
MgSO.sub.4.7H.sub.2O 0.5 gm Yeast extract 1.0 gm Soy peptone 5.0 gm
Trace minerals 10.0 ml Dextrose 30.0 gm Deionized water 1.0 L pH
adjusted to 9.0 before sterilization Trace Minerals Solution 5074H
FeSO.sub.4.7H.sub.2O 10.0 mg ZnSO.sub.4.7H.sub.2O 8.8 mg
CuSO.sub.4.5H.sub.2O 15.0 mg MnSO.sub.4.H.sub.2O 7.6 mg
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.sub.2O 10.0 mg Deionized water
1.0 L
Procedure
[0313] Inoculum:
[0314] A 500 ml Erlenmeyer flask containing 100 ml of the above
medium was inoculated with Diplodia gossypina ATCC 10936, using a
stock slant culture. 10 Ml of sterile 50% dextrose were added and
the flask was incubated for 72 hours at 30.degree. C. and 150
rpm.
[0315] Production:
[0316] 500 Ml of the above medium was added to a 2.8 liter Fembach
flask and sterilized for 20 minutes at 121.degree. C. The 72 hour
inoculum was chopped in a sterile blender jar, and 10 ml were added
along with 50 ml of sterile 50% dextrose to the Fembach flask,
followed by stationary incubation at 30.degree. C.
Results
[0317] After 10 days of incubation, 402 ml of culture broth were
recovered, having a titer of 1.2 gram/liter jasmonic acid as
determined by HPLC.
EXAMPLE V
Production of Jasmonic Acid in Aseptic Stationary Tray Culture
[0318] Reactions:
15 80 81 and 82 83 Medium Medium 5059H KH.sub.2PO.sub.4 1.0 gm
MgSO.sub.4.7H.sub.2O 0.5 gm Yeast extract 10.0 gm Soy peptone 5.0
gm Trace minerals 10.0 ml Dextrose 30.0 gm Deionized water 1.0 L pH
adjusted to 8.5 before sterilization Trace Minerals Solution 5059H
FeSO.sub.4.7H.sub.2O 10.0 mg ZnSO.sub.47H.sub.2O 8.8 mg
CuSO.sub.4.5H.sub.2O 15.0 mg MnSO.sub.4.H.sub.2O 7.6 mg
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.su- b.2O 10.0 mg Deionized
water 1.0 L
Procedure
[0319] Inoculum:
[0320] A 500 ml Erlenmeyer flask containing 100 ml of the above
medium was inoculated with Diplodia gossypina ATCC 10936, using a
stock slant culture. 10 Ml of sterile 50% dextrose were added and
the flask was incubated for 72 hours at 30.degree. C. and 150
rpm.
[0321] Production:
[0322] 3 Liters of sterile broth, adjusted to pH 8.5 before
sterilization, were inoculated with 10 ml of a 72 hour chopped
inoculum. 300 Ml of sterile 50% dextrose were added, the mixture
poured into a sterile plastic tray (approximately 11 inches long,
10 inches wide and 5 inches deep) and covered with a
cheesecloth/cotton screen to maintain pure culture conditions. The
tray culture was incubated stationary for 14 days at 3 1.5.degree.
C.
Results
[0323] The culture broth had a titer of 0.71 gram/liter after 10
days of incubation. The final recovered broth (2.14 liters, 14
days) had a titer of 1.24 gramliter as deternined by HPLC.
EXAMPLE VI
Tray Culture Production of Jasmonic Acid
[0324] Reactions:
16 84 85 and 86 87 Medium Medium 5059H KH.sub.2PO.sub.4 1.0 gm
MgSO.sub.4.7H.sub.2O 0.5 gm Yeast extract 1.0 gm Soy peptone 5.0 gm
Trace minerals 10.0 ml Dextrose 30.0 gm Deionized water 1.0 L pH
adjusted to 9 before sterilization
[0325] Chloramphenicol was added after sterilization to help
protect against contamination.
17 Trace Minerals Solution 5059H FeSO.sub.4.7H.sub.2O 10.0 mg
ZnSO.sub.4.7H.sub.2O 8.8 mg CuSO.sub.4.5H.sub.2O 15.0 mg
MnSO.sub.4.H.sub.2O 7.6 mg
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.sub.2O 10.0 mg Deionized water
1.0 L
Procedure
[0326] Inoculum:
[0327] A 72 hour inoculum was prepared in 2.8 liter Fembach flasks,
each flask containing 100 ml. The flasks were incubated at
30.degree. C. and 150 rpm.
[0328] Production:
[0329] A total of 540 liters of broth was prepared in a 300 gallon
fermenter. The pH was adjusted to 9.0, and the broth was sterilized
for 30 minutes at 121.degree. C. After cooling to 30.degree. C.,
chloramphenicol (36 grams dissolved in 600 ml ethanol), 60 liters
of sterile 50% dextrose and 2,307 grams chopped inoculum (weight
adjusted for a standard 45% packed cell volume) were added. After
mixing for 5-10 minutes, the inoculated broth was transferred to
large plastic trays (approximately 20 inches long, 17 inches wide
and 5 inches deep), placed on shelves in an
environmentally-controlled room. Approximately 6 liters of
inoculated broth was added to each tray (total of 100 trays), the
trays covered with aluminum foil and incubated stationary at
31-32.degree. C. for 14 days. At the conclusion of incubation, the
mycelial mats were filtered off, the broth was transferred to the
300 gallon fermenter and sterilized for 20 minutes at 121.degree.
C. The broth was cooled to room temperature, acidified to pH 4, and
the jasmonic acid was recovered by adsorption/elution on a nonionic
resin.
Results
[0330] The bulked broth (approximately 380 liters) from the 100
tray cultures had a titer of 0.895 gram/liter jasmonic acid as
determined by HPLC.
EXAMPLE VII
Production of Jasmonic Acid by Combined Surface-submerged
Fermentation
[0331] Reactions:
18 88 89 and 90 91 Medium Medium 5059H KH.sub.2PO.sub.4 1.0 gm
MgSO.sub.4.7H.sub.2O 0.5 gm Yeast extract 1.0 gm Soy peptone 5.0 gm
Trace minerals 10.0 ml Dextrose 30.0 gm Deionized water 1.0 L pH
adjusted to 9 before sterilization
[0332] Chloramphenicol was added after sterilization to help
protect against contamination.
19 Trace Minerals Solution 5059H FeSO.sub.4.7H.sub.2O 10.0 mg
ZnSO.sub.4.7H.sub.2O 8.8 mg CuSO.sub.4.5H.sub.2O 15.0 mg
MnSO.sub.4.4H.sub.2O 7.6 mg
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.sub.2O 10.0 mg Deionized water
1.0 L
Procedure
[0333] Inoculum:
[0334] A 500 ml Erlenmeyer flask containing 100 ml of the above
medium was inoculated with Diplodia gossypina ATCC 10936, using a
stock slant culture. 10 Ml of sterile 50% dextrose were added and
the flask was incubated for 72 hours at 30.degree. C. and 150
rpm.
[0335] Production:
[0336] A single coil of stainless steel wire cloth, having
approximately 3/8 inch.times.3/8 inch openings, was attached
vertically to the baffles of a 150 liter fermenter. Four circular
wire cloth screens (same opening size) were attached to the
vertical coil in horizontal positions such that the lowest screen
was about one inch above the broth surface, and the other screens
were separated by about one inch space between each screen. An
opening in the center of each screen permitted freedom for
agitation, using one impeller.
[0337] 60 Liters of broth were made up and transferred to the
fermenter, the pH adjusted to 9.0 and the broth sterilized at
121.degree. C. for 30 minutes. After cooling to 30.degree. C., 2.4
liters of chopped 72 hour inoculum, chloramphenicol (6.0 grams
dissolved in 60 ml of ethanol) and 1.2 liters of sterile 50%
dextrose were added. Sterile 50% dextrose solution and sterile
deionized water were added as needed during the 19 day incubation
at 30.degree. C. Aeration was provided and agitation was engaged
periodically to prevent the developing mycelium attached to the
screens from drying out.
Results
[0338] At the conclusion of the incubation, approximately 31 liters
of broth were recovered, having a titer of 185 mg/liter jasmonic
acid as determined by HPLC.
EXAMPLE VIII
Production of Jasmonic Acid Using Plastic Biofilter Supports
[0339] Reactions:
20 92 93 and 94 95 Medium Medium 5059H KH.sub.2PO.sub.4 1.0 gm
MgSO.sub.4.7H.sub.2O 0.5 gm Yeast extract 1.0 gm Soy peptone 5.0 gm
Trace minerals 10.0 ml Dextrose 30.0 gm Deionized water 1.0 L pH
adjusted to 9 before sterilization
[0340] Chloramphenicol was added after sterilization to help
protect against contamination.
21 Trace Minerals Solution 5059H FeSO.sub.4.7H.sub.2O 10.0 mg
ZnSO.sub.4.7H.sub.2O 8.8 mg CuSO.sub.4.5H.sub.2O 15.0 mg
MnSO.sub.4.H.sub.2O 7.6 mg
(NH.sub.4).sub.6Mo.sub.7O.sub.24.4H.sub.2O 10.0 mg Deionized water
1.0 L
Procedure
[0341] Inoculum:
[0342] A 500 ml Erlenmeyer flask containing 100 ml of the above
medium was inoculated with Diplodia gossypina ATCC 10936, using a
stock slant culture. 10 Ml of sterile 50% dextrose were added and
the flask was incubated for 72 hours at 30.degree. C. and 150 rpm.
The pH of the broth was adjusted to 9.0 before sterilization.
[0343] Production:
[0344] A 130 liter fermenter, equipped with a spray ball attached
to the center of the headplate, was filled 2/3 full with Jaeger
Tripack biofilter 2 inch diameter supports. 30 Liters of broth was
prepared and, after the pH was adjusted to 9.0, was charged to the
fermenter. A recirculation loop was provided for the purposes of
broth sterilization, for periodic broth recirculation (through the
spray nozzle) during incubation to replenish nutrients to the
developing mycelium attached to the biosupports and to prevent
drying out. Sterilization was carried out by continuous
recirculation for 30 minutes at 121.degree. C. After cooling to
30.degree. C., 30 ml of chopped inoculum chloramphenical (3 grams
dissolved in 30 ml of ethanol) and 3 liters of sterile 50% dextrose
were added. Incubation was carried out at 30.degree. C. without
agitation, and periodic recirculation of the inoculated broth was
performed. Aeration was provided, and sterile 50% dextrose solution
and deionized water were added as needed.
Results
[0345] Based on the original 30 liters of staring broth, the batch
yielded 146 mg/liter jasmonic acid as determined by HPLC.
EXAMPLE IX
Production of Jasmonic Acid in Shake Flask Culture
[0346] Reactions:
22 96 97 and 98 99 Medium: Buffered Minimal Salts Medium (bMS)
NaNO.sub.3 0.5 gm/L KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5
gm/L MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L
FeSO.sub.4-7H.sub.2O 1.0 mg/L TASTONE .RTM. 900 0.5 gm/L
[0347] Following sterilization of bMS medium at 120.degree. C. and
15 psi 60 g/l of 50% sterile glucose solution was added.
Procedure
[0348] Inoculum:
[0349] A 500 ml Erlenmeyer flask containing 100 ml of sterile bMS
medium was inoculated with 2 ml of a frozen chopped cell culture of
Diplodia gossypina ATCC 10936. 6 Ml of sterile 50% dextrose were
added and the flask was incubated for 3 days at 28.degree. C. and
200 rpm. Following 3 days of incubation, the culture was chopped
for 1 minute in a sterile Waring Blender and this was used as
inoculum.
[0350] Production:
[0351] 1 Ml of inoculum was added to 100 ml of sterile bMS medium
in a 500 ml Erlenmeyer flask. 6 M1 of sterile 50% dextrose were
added and the flask was incubated at 28.degree. C. and 200 rpm. The
production of jasmonic acid in the culture was monitored by TLC and
HPLC analyses.
Results
[0352] Diplodia gossypina ATCC 10936 produced 1.2 gram/liter
jasmonic acid after 7 days of incubation.
EXAMPLE X
Production of Jasmonic Acid in Fernbach Flask Submerged Culture
[0353] Reactions:
23 100 101 and 102 103 Medium: Buffered Minimal Salts Medium (bMS)
NaNO.sub.3 2.0 gm/L KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5
gm/L MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L
FeSO.sub.4-7H.sub.2O 1.0 mg/L TASTONE .RTM. 900 0.5 gm/L
[0354] After sterilization of bMS medium at 120.degree. C. and 15
psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0355] Inoculum:
[0356] A 500 ml Erlenmeyer flask containing 100 ml of sterile bMS
medium was inoculated with 2 ml of a frozen chopped cell culture of
Diplodia gossypina ATCC 10936. 6 Ml of sterile 50% dextrose were
added and the flask was incubated for 3 days at 28.degree. C. and
200 rpm. Following 3 day of incubation, the culture was chopped for
1 minute in a sterile Waring Blender and this was used as
inoculum.
[0357] Production:
[0358] 5 Ml of inoculum were added to 500 ml of sterile bMS medium
in a Fembach flask. 30 Ml of sterile 50% dextrose were added and
the flask was incubated at 28.degree. C. and 150 rpm. The
production ofjasmonic acid was monitored by TLC and HPLC
analyses.
Results
[0359] Diplodia gossypina ATCC 10936 produced jasmonic acid at a
titer of 0.9 gram/liter after 8 days of incubation.
EXAMPLE XI
Production of Jasmonic Acid in Laboratory Fermenters
[0360] Reactions:
24 104 105 and 106 107 Medium: Buffered Minimal Salts Medium (bMS)
NaNO.sub.3 1.0 gm/L KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5
gm/L MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L
FeSO.sub.4-7H.sub.2O 1.0 mg/L TASTONE .RTM. 900 0.5 gm/L
[0361] Following sterilization of bMS medium at 120.degree. C. and
15 psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0362] Inoculum:
[0363] A 500 ml Erlenmeyer flask containing 100 ml of sterile bMS
medium was inoculated with 2 ml of a frozen chopped cell culture of
Diplodia gossypina ATCC 10936. 6 Ml of sterile 50% dextrose were
added and the flask was incubated for 3 days at 28.degree. C. and
200 rpm. Following 3 days of incubation, the inoculum was chopped
for 1 minute in a sterile Waring Blender. 1 Ml of inoculum and 6 ml
of sterile 50% dextrose were added to 100 ml of sterile bMS medium.
After another 3 days of incubation, the whole culture was used as
an inoculum for a laboratory 10 liter fermenter.
[0364] Production:
[0365] 10 Liters of bMS medium containing 1 gram/liter of
NaNO.sub.3 was added to a 14 liter laboratory fermenter and
sterilized for 30 minutes at 121.degree. C. The above inoculum and
600 grams of sterile 50% dextrose were added to a fermenter. The
fermenter was run at 28.degree. C. and 1,000 rpm with aeration at 1
VVM and 15 psi back pressure. The pH of fernenter was maintained at
6.0 by addition of 25% NaOH. After 4 days of incubation, an
additional 0.5 gram/liter aliquot of NaNO.sub.3 was added. The
concentration of glucose in the broth was monitored and additional
sterile 50% dextrose was added to prevent glucose depletion. The
production of jasmonic acid in the broth was monitored by TLC and
HPLC analyses.
Results
[0366] After 13 days of incubation, Diplodia gossypina ATCC 10936
produced jasmonic acid at a titer of 0.85 gram/liter.
EXAMPLE XII
Production of Jasmonic Acid In Submerged 100-l Pilot Plant
Fermenters
[0367] Reactions:
25 108 109 and 110 111 Medium: Buffered Minimal Salts Medium (bMS)
NaNO.sub.3 0.5 gm/IL KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5
gm/L MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L
FeSO.sub.4-7H.sub.2O 1.0 mg/L TASTONE .RTM. 900 0.5 gm/L
[0368] Following sterilization of bMS medium at 120.degree. C. and
15 psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0369] Inoculum:
[0370] A 500 ml Erlenmeyer flask containing 100 ml of sterile bMS
medium was inoculated with 2 ml of a-frozen chopped cell culture of
Diplodia gossypina ATCC 10936. 6 Ml of sterile 50% dextrose were
added and the flask was incubated for 3 days at 28.degree. C. and
200 rpm. Following 3 days of incubation, the inoculum was chopped
for 1 minute in a sterile Waring Blender. 5 Ml of the inoculum and
30 ml of sterile 50% dextrose were added to each of two Fembach
flasks containing 500 ml of sterile bMS medium. The flasks were
incubated at 28.degree. C. and 150 rpm for 72 hours.
[0371] Production:
[0372] 100 Liters of bMS containing 1 gram/liter of NaNO.sub.3 were
charged to a 150 liter pilot plant fermenter and sterilized for 30
minutes at 121 C. 1,000 Ml of whole inoculum and 600 grams of
sterile 50% dextrose were added to the fermenter. The fermenter was
run at 28.degree. C. and 450 rpm with aeration at I VVM and 15 psi
back pressure. The pH of fermenter was maintained at 6.0 by
addition of 25% NaOH. After 4 days of incubation, an additional 0.5
gram/liter aliquot of NaNO.sub.3 was added. The concentration of
glucose in the broth was monitored and additional sterile 50%
dextrose was added to prevent glucose depletion. The production of
jasmonic acid in cell culture was monitored by TLC and HPLC
analyses.
Results
[0373] After 9 days of incubation, Diplodia gossypina ATCC 10936
produced jasmonic acid at a titer of 0.85 gram/liter.
EXAMPLE XIII
Effect of 10-Oxo-8-trans-decenoic Acid on the Production of
Jasmonic Acid in Fernbach Flask Submerged Culture
[0374] Reactions:
26 112 113 and 114 115 Medium: Buffered Minimal Salts Medium (bMS)
NaNO.sub.3 0.5 gm/L KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5
gm/L MgSO.sub.4-7H.sub.2O 2.0 gm/L KCl 0.5 gm/L
FeSO.sub.4-7H.sub.2O 1.0 mg/L TASTONE .RTM. 900 0.5 gm/L
[0375] Following sterilization of bMS medium at 120.degree. C. and
15 psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0376] Inoculum:
[0377] A 500 ml Erlenmeyer flask containing 100 ml of sterile bMS
medium was inoculated with 2 ml of a frozen chopped cell culture of
Diplodia gossypina ATCC 10936. 6 Ml of sterile 50% dextrose were
added and the flask was incubated for 3 days at 28.degree. C. and
200 rpm. Following 3 days of incubation, the culture was chopped
for 1 minute in a sterile Waring Blender and this was used as
inoculum.
[0378] Production:
[0379] 5 Ml of inoculum and 30 ml of sterile 50% dextrose were
added to a Fernbach flask containing 500 ml of sterilize bMS
medium. A stock solution of 50% 10-oxo-decenoic acid (10-ODA) in
ethanol was prepared and dispensed to the flasks at a final
concentration of either 1 or 10 ppm. The cultures were incubated at
28.degree. C. and 150 rpm. The production ofjasmonic acid was
monitored by TLC and HPLC analyses.
Results
[0380]
27 Number Flasks 3 Days 6 Days 1 Control 106 ppm 249 ppm 2 1 ppm
ODA 197 ppm 538 ppm 3 10 ppm ODA 279 ppm 630 ppm
EXAMPLE XIV
Effect of 10-oxo-8-trans-decenoic Acid on the Production of
Jasmonic Acid in A Submerged 10-l of Laboratory Fermenter
[0381] Reactions:
28 116 and 117 Medium: Buffered Minimal Salts Medium (bMS)
NaNO.sub.3 2.0 gm/L KH.sub.2PO.sub.4 2.0 gm/L K.sub.2HPO.sub.4 0.5
gm/L MgSO.sub.4--7H.sub.2O 2.0 gm/L KCl 0.5 gm/L
FeSO.sub.4--7H.sub.2O 1.0 mg/L TASTONE .RTM. 900 0.5 gm/L
[0382] After sterilization of bMS medium at 120.degree. C. and 15
psi, 60 g/l of 50% sterile glucose solution was added.
Procedure
[0383] Inoculum:
[0384] A 500 ml Erlenmeyer flask containing 100 ml of sterile bMS
medium was inoculated with 2 ml of a frozen chopped cell culture of
Diplodia gossypina ATCC 10936. 6 Ml of sterile 50% dextrose were
added and the flask was incubated for 3 days at 28.degree. C. and
200 rpm. Following 3 days of incubation, the inoculum was chopped
for 1 minute in a sterile Waring Blender. 1 Ml of inoculum and 6 ml
of sterile 50% dextrose were added to 100 ml of sterile bMS medium.
After another 3 days of incubation, the whole culture was used as
an inoculum for a laboratory 10 liter fermenter.
[0385] Production:
[0386] 10 Liters of bMS containing 1.0 grams/liter of NaNO.sub.3
were charged to a 14 liter laboratory fermenter and sterilized for
30 minutes at 121.degree. C. 100 Ml of inoculum lated and 600 grams
of sterile 50% dextrose were added to the fermenter. A stock
solution of 10-ODA was prepared as in Example XIII and added to the
fermenter at a final concentration of 10 ppm in the broth. The
fermenter was run at 28.degree. C. and 1,000 rpm with aeration at 1
VVM and 15 psi back pressure. The pH of fermenter was maintained at
6.0 by the addition of 25% NaOH. After 4 days of incubation, an
additional 0.5 grams/liter aliquot of NaNO.sub.3 was added. The
concentration of glucose in the broth was monitored and additional
sterile 50% dextrose was added to prevent glucose depletion. The
production of jasmonic acid was monitored by TLC and HPLC
analyses.
Results
[0387] After 11 days of incubation, Diplodia gossypina ATCC 10936
produced jasmonic acid at a titer of 1.5 grams/liter.
EXAMPLE XV
Production of Natural "EPI" Meteyl Jasmonate-preparation and
Purification from Natural Jasmonic Acid Fermentation Broth
Extract
[0388] Reactions: 118
29 Purification Scheme Step Description 1.0 119 2.0 Crude RF
Jasmonic Acid optional topping 240 grams 120 3L aqueous solution
3.0 121 4.0 aqueous solution H.sub.3PO.sub.4 re-acidification to pH
2 4.1 122 4.2 123 5.0 Purified Jasmonic Acid (615 grams) 2460 grams
Nat Methanol 5.1 6.0 7.0 8.0 124
Description of Process Steps
[0389] 1.0 The rush-over of the crude topped extract is carried out
under best vacuum to a final pot temperature of 260 C. At higher
temperatures, there is thermal breakdown and some loss of vacuum.
Most of the charge remains as "residue." A portion of this residue
was dissolved in ethyl acetate and extracted with aqueous base.
Re-acidification and GLC analysis indicated that there was no
jasmonic acid left remaining in the residue.
[0390] 2.0. The extract is stirred and 3 liters of 6% sodium
carbonate solution to neutralize the sodium jasmonate. The pH is
adjusted to 9.
[0391] 3.0. The basic solution is extracted three times with
hexane. This removes non-acidic impurities as well as significant
sulfurous odor bodies. At lower pH, more material is removed.
[0392] The hexane extract contains approximately 160 grams of
organic material. The major component is identified as a lactone
having the structure: 125
[0393] This lactone is derived from the alcohol according to the
reaction: 126
[0394] 4.0. Approximately 225 grams of 85% phosphoric acid is
required to adjust the acidity to pH=2.
[0395] 5.0 Natural jasmonic acid having the structures: 127
[0396] is charged to an autoclave along with 4 times its weight of
natural methanol. The autoclave is sealed and operated for a period
of 12 hours at 150.degree. C. The final composition is
approximately 70:30 (weight:weight) methyl jasmonate:jasmonic acid.
The autoclave pressure is 220 psig.
[0397] 6.0 The autoclave is opened and the methanol stripped from
the resulting product. The pH of the product is adjusted to 9.
[0398] 7.0 Ethyl acetate is used to extract the resulting methyl
jasmonate. The ethyl acetate is stripped at a temperature below
60.degree. C. at atmospheric pressure. The resulting methyl
jasmonate is then fractionally distilled, using a 12 inch.times.1.5
inch Goodloe packed column. The first fraction is distilled at a
reflux ratio of 4:1, and the remainder of the fractions are
distilled at reflux ratio of 1:1. The conditions of distillation
are: 125.degree. C.; vapor temperature at 1 mm/Hg pressure.
[0399] 8.0 The methyl jasmonate is redistilled at a reflux ratio of
4:1 at 125.degree. C. and 1 mm/Hg pressure.
EXAMPLE XVI
[0400] A white chocolate raspberry flavor was prepared for addition
to nonfat yogurt containing aspartame.
[0401] The following flavor composition was prepared:
30 Ingredients Parts by Weight Jasmonic acid prepared according to
Example VI 15 Natural cocoa extract 28 Natural raspberry extract 30
3-Phenyl-4-pentenal 8
[0402] At the rate of 8 ppm, the above-mentioned mixture was added
to natural nonfat yogurt containing aspartame, cultured,
pasturized, grade A nonfat milk, modified cornstarch, whey protein
and gelatin, and active cultures with L. acidophilus.
[0403] On mixing the flavor with the natural yogurt, the resulting
product has a natural raspberry, fresh, "just-picked," "seedy"
taste with floral topnote.
[0404] A second sample was flavored without the jasmonic acid
present, and the resulting flavor did not have the "fresh-just
picked-seedy" quality.
EXAMPLE XVII
Jasmine Perfume Formulation
[0405] The following mixture is prepared:
31 Parts by Ingredients Weight Jasmonic acid product prepared
according to Example XIV 50 Methyl jasmonate product prepared
according to Example 50 XV Orange oil 20 Bergamot oil 20 Neroli oil
20 .gamma.-Methyl ionone 20 1-Acetyl-2,5,5-trimethyl cycloheptane
45
[0406] The products of Examples XIV and XV impart to this jasmine
perfume formulation powerful, long lasting jasmine,
floral-herbaceous aromas with sweet-herbaceous, green-woody
topnotes. Accordingly, the perfume composition of Example XVII can
be described as "jasmine with floral-herbaceous undertones and
sweet-herbaceous, green-woody topnotes."
EXAMPLE XVIII
Preparation of Soap Compositions
[0407] 100 Grams of soap chips are produced according to Example V
of U.S. Pat. No. 4,058,487 issued on Nov. 5, 1997, the
specification for which is incorporated herein by reference, as
follows:
[0408] The sodium salt of an equal mixture of C.sub.10-C.sub.14
alkane sulfonate (95% active), 40 lbs, is dissolved in a mixture of
80 lbs of anhydrous isopropanol and 125 lbs of deionized water at
150.degree. F. In this mixture is dissolved 10 lbs of partially
hydrogenated coconut oil fatty acids and 15 lbs of sodium
mono-C.sub.14 alkyl maleate, and the pH of this solution is
adjusted to 6.0 by the addition of a small amount of 50% aqueous
solution of sodium hydroxide. The isopropanol is distilled off and
the remaining aqueous solution is drum dried. The resulting solid
actives are then blended in a chip mixture with 10 lbs of water,
0.2 lbs of titanium hydroxide and 0.7 lbs of one of the perfume
ingredients set forth in Table V, infra.
32TABLE V Ingredients Fragrance Profile Jasmonic acid isomer having
the structure: A powerful, long lasting jasmine, floral- herbaceous
aroma with sweet-herbaceous, green-woody topnotes. 128 (optical
rotation: +58.degree.) Isomer having the structure; A powerful,
long lasting jasmine, floral- herbaceous aroma with
sweet-herbaceous, green-woody topnotes. 129 (optical rotation:
+58.degree.). Isomer having the structure: A powerful, long lasting
jasmine, floral- herbaceous aroma with sweet-herbaceous,
green-woody topnotes. 130 (optical rotation: +58.degree.). Isomer
having the structure: A powerful, long lasting jasmine, floral-
herbaceous aroma with sweet-herbaceous, green-woody topnotes. 131
(optical rotation: +58.degree.). Perfume composition of Example
XVII. Jasmine with floral, herbaceous undertones and sweet,
herbaceous, green, woody topnotes.
EXAMPLE XIX
[0409] Preparation of Detergent Composition
[0410] A total of 100 grams of a detergent powder prepared
according to U.S. Pat. No. 4,058,472 (the specification for which
is incorporated by reference herein) and containing 5% by weight of
the sodium salts of a mixture of sulfonated C.sub.14-C.sub.18 alkyl
catechol as a surface active component, the mixture being 60 parts
by weight of mono- C.sub.14-C.sub.18 alkyl catechol and 40 parts by
weight of di-C.sub.14-C.sub.18 catechol, 35% sodium
tetrapyrophosphate, 30% sodium silicate, 20% of sodium carbonate,
3% of sodium carboxymethyl cellulose and 7% of starch is mixed with
0.15 grams individually with each of the aroma ingredients set
forth in Table V of Example XVIII until a substantially homogeneous
composition is obtained. Each of the compositions has an excellent
aroma as set forth in Table V of Example XVIII.
EXAMPLE XX
Preparation of a Cosmetic Powder Composition
[0411] A cosmetic powder is prepared by mixing in a ball mill, 100
grams of talcum powder with 0.25 grains of each of the perfume
materials of Table V of Example XVIII. Each of the powders has an
excellent aroma as set forth in Table V of Example XVIII.
EXAMPLE XXI
Perfumed Liquid Detergent
[0412] Concentrated liquid detergents with aromas as set forth in
Table V of Example XVIII are prepared by adding 0.10%, 0.15% and
0.20% of each of the ingredients set forth in Table V of Example
XVIII. They are prepared by adding and homogeneously mixing the
appropriate quantity of perfume substance of Table V of Example
XVIII in the liquid detergent. The detergents individually possess
aromas as set forth in Table V of Example XVIIL the intensity
increasing with greater concentrations of perfume substance set
forth in Table V of Example XVIII.
EXAMPLE XXII
Preparation of a Cologne and Handkerchief Perfume
[0413] Each of the ingredients of Table V of Example XVIII is
incorporated individually into colognes of several strengths at
concentrations of 2.0%, 2.5%, 3.0%, 3.5%, 4.0% and 5.0% in 75%,
80%, 85%, 90% and 95% aqueous ethanol; and into several
concentrations of handkerchief perfumes at the rate of 15%, 20% and
25% (in 80%, 85%, 90% and 95% aqueous ethanol). Distinct and
definite aromas as set forth in Table V of Example XVIII are
imparted to the colognes and to the handkerchief perfumes at the
several concentrations set forth above.
EXAMPLE XXIII
Preparation of Soap Compositions
[0414] 100 Grams of soap chips (IVORY.RTM., produced by the Procter
& Gamble Company of Cincinnati, Ohio) are admixed with 1 gram
of each of the substances set forth in Table V of Example XVIII,
supra, until homogeneous compositions are heated under 3
atmospheres pressure at 180.degree. C. for a period of 3 hours, and
the resulting liquids are placed into soap molds. The resulting
soap cakes, on cooling, manifest excellent aromas as set forth in
Table V of Example XVIII.
EXAMPLE XXIV
Preparation of Solid Detergent Compositions
[0415] Detergents are prepared from the following ingredients
according to Example I of Canadian Patent No. 1,007,948, the
specification for which is incorporated by reference herein:
33 Parts by Ingredients Weight NEODOL .RTM. 45-11 (a
C.sub.14-C.sub.15 alcohol ethoxylated with 12 11 moles of ethylene
oxide) Sodium carbonate 55 Sodium citrate 20 Sodium sulfate, water
brighteners q.s.
[0416] This detergent is a "phosphate-free" detergent. A total of
100 grams of said detergent is admixed with 0.10, 0.15, 0.20 and
0.25 grams of each of the substances set forth in Table V of
Example XVIII, supra. Each of the detergent samples has an
excellent aroma as indicated in Table V of Example XVIII.
EXAMPLE XXV
Preparation of Drier-added Fabric Softener Article
[0417] Utilizing the procedure of Example I at column 15 of U.S.
Pat. No. 3,632,396, the specification for which is incorporated by
reference herein, a non-woven cloth substrate useful as a
drier-added fabric softening article of manufacture is prepared
wherein the substrate, substrate coating, outer coating and the
perfume material are as follows:
[0418] 1. a water "dissolvable" paper ("Dissolvo Paper") as the
substrate;
[0419] 2. ADOGEN.RTM. 448 (melting point about 140.degree. F.) as
the first substrate coating; and
[0420] 3 . an outer coating having the following formulation
(melting point about 150.degree. F.):
[0421] 57% C.sub.20-C.sub.22 HAPS;
[0422] 22% isopropyl alcohol;
[0423] 20% antistatic agent; and
[0424] 1% of one of the perfumery substances set forth in Table V
of Example XVIII, supra.
[0425] Fabric softening compositions containing the substances as
set forth in Table V of Example XVIII, supra, essentially consist
of a substrate having a weight of about 3 grams per 100 square
inches; a substrate coating weighing about 1.85 grams per 100
square inches of substrate; and an outer coating weighing about 1.5
grams per 100 square inches of substrate are prepared, thereby
providing a total aromatized substrate and outer coating weight
ratio of about 1:1 by weight of the substrate.
[0426] The aromas as set forth in Table V of Example XVIIL supra,
are imparted in a pleasant manner to the headspace in a drier on
operation thereof, using the said drier-added fabric softening
non-woven fabric by adding to the drying cycle.
[0427] As stated above in the case of fabric softener articles, the
entire U.S. Pat. No. 3,632,396 is incorporated by reference herein.
Thus, all of the articles of U.S. Pat. No. 3,632,396, acting as
fabric softening articles in said U.S. Pat. No. 3,632,396, may be
perfumed in their outer coating with from 0.25% up to 5% by weight
of each of the perfuming substances of Table V of Example XVIII,
supra.
EXAMPLE XXVI
Hair Preparation
[0428] A "soft-feel, good-hold" hair spray is produced containing
the following ingredients:
34 Ingredients Parts by Weight Polyvinylpyrollidone/vinyl acetate
"E-735 Copolymer" 4.00 manufactured by the GAF Corporation of New
York, NY Anhydrous ethanol 70.90 Dioctyl sebecate 0.05 Benzyl
alcohol 0.05 "Propellant A-46" manufactured by the GAF 24.95
Corporation of New York, NY Fragrance ingredient as set forth in
Table V 0.05 of Example XVIII, supra
[0429] The PVP/VA copolymers are first dissolved in alcohol and all
other ingredients are added until uniform. The propellant is then
pressurized and used as an aerosol. The resulting hair sprays each
have pleasant aromas as set forth in Table V of Example XVIII,
supra.
EXAMPLE XXVII
Scouring Cleanser Composition
[0430] A scouring cleanser composition is prepared in accordance
with Example I at columns 11 and 12 of U.S. Pat. No. 4,193,888
issued on Mar. 18, 1980, the specification for which is
incorporated by reference herein. To this composition, the
substances set forth in Table V of Example XVIII, supra, are added
at the level of 0.25% as set forth in the table in said Example I
of U.S. Pat. No. 4,193,888, yielding an aroma on using said
cleanser in ordinary circumstances which is quite pleasant and
described in Table V of Example XVIII, supra.
EXAMPLE XXVIII
[0431] A fabric softening article prepared substantially as set
forth in Example VIII of Canadian Patent No. 1,069,260, the
specification for which is incorporated by reference herein, is
prepared containing 0.21% by weight of a perfuming substance as set
forth in Table V of Example XVIII, supra, and yielding, on use in a
drier, a faint aroma as set forth in Table V of Example XVIII,
supra.
EXAMPLE XXIX
Tobacco Flavor Formulations
[0432] Cigarettes are produced using the following
formulations:
35 Ingredients Parts by Weight Bright 40.1 Burley 24.9 Maryland 1.1
Turkish 11.6 Stem (flue cured) 14.2 Glycerine 2.8 H.sub.2O 5.3
[0433] At the rate of 0.2%, the following tobacco formulation is
applied to all of the cigarettes produced with the above tobacco
formulation:
36 Parts by Ingredients Weight Ethyl butyrate 50 Ethyl valerate 50
Maltol 20 Cocoa extract 20 Coffee extract 20 Ethyl alcohol (95%) 45
H.sub.2O 41.900
[0434] To portions of 50% of the cigarettes at levels of 10 and 20
ppm, the jasmonic acid-containing composition of Example VI is
added. These cigarettes are hereinafter called "experimental"
cigarettes. The cigarettes without the jasmonic acid composition
are hereinafter called "control" cigarettes. The control and
experimental cigarettes are then evaluated by paired comparison and
the results are as follows:
[0435] (a) in aroma, the experimental cigarettes are all found to
be more aromatic with Turkish tobacco-like nuances; and
[0436] (b) in smoke flavor, the experimental cigarettes are all
found to be more aromatic, more sweet with Turkish tobacco,
oriental-like nuances than the control cigarettes.
[0437] The experimental cigarettes containing the mixture of
jasmonic acids are found to be fruity and floral and have pleasant,
aesthetically pleasing fruity and floral notes, in addition.
* * * * *