Title of Invention

A PROCESS FOR THE PRODUCTION OF SAFFRON PIGMENTS AND FLAVOUR CONCENTRATE

Abstract A process for the isolation of saffron pigments and flavour concentrate by mixing saffron with one food-grade solvent or a combination of more than one food graded-solvents, macerating and agitating the mixture with continuance protection from light condition at a temperature in the range of 5°C to 100°C, centrifuging/filtering the above said macerated mixture to remove undesirable fibrous plant material, cooling the centrifuge immediately to a temperature in the range of -30"C to -20°C under inert atmosphere at a pH in the range of 4 to 9, lyophilizing the cooled centrifuge under reduced pressure to obtain crude material, isolating the concentrate by column chromatography using silica gel method and obtaining brilliant orange color shining saffron pigments and flavour concentrate with recovery of 95%.
Full Text A PROCESS FOR THE ISOLATION OF SAFFRON PIGMENT AND FLAVOUR CONCENTRATE Field of the present invention
The present invention relates to an efficient method for the extraction of high stability, superior quality, value added, standardized, ready-to-use saffron pigments and flavour concentrate with recovery of about 95% and use of the saffron pigments and flavour concentrate of range between 0.05 to 3% in food, pharmaceutical, and allied industries for flavour, and color. Background Art
I he current trend in food colorants has inc easmgly centered on naturally occurring pigments because of consumer preferences. Crocm. the digentiobioside of crocetin. and other crocetin glycosides namely crocetin gentiobioside glucoside. crocetin diglucoside and others are probably the only true water-soluble natural carotenoid pigments and are the constituent of saffron, the world's most expensive spice. (Basker D, Negbi M 1983 Uses of saffron. I-'.a> Hoi 37. 228- 236.) and of cape jasmine fruit [(Gardenia j'axminoidix Ellis, Ichi T. Katayema 1. Toda M 1993 ( hanges in carotenoid pattern during development of gardenia fruit. Okayuma Daiguku\cjftikuhn Gakujutsu Hokoku (Japan) N2. 9-15; ( Chem Abstr 1994, 121 : 104261u); Kamikura M. Naka/ato K. 1985 Comparison of natural yellow colors extracted from saffron. Crocus xaimix and gardenia fruit, Gardenia jasminoides. Eisei Shikensho Hokoku (Japan) 103. 157-160(Chem Abstr 1986 105: 75885 n)]. Attempts for the production of crocin pigment in cultured cells of G. jitsminoidis is also underway [(Nawa Y. Ohtali 1. 1992 Induction of Callus from flesh of Gardenia jasrninmJe.x Ellis fruit and formation of yellow pigment in the callus. Biochem 56. 1732-1736: Kim S H. Park Y G, Lu Y H 1991 Suspension i ulture of Gardenia jasminoides Ellis cell for production of yellow pigment. ./. Microbial Bioiechnol 1 142-149: K( )hda H 1991 Production of useful pigments in cultured cells of Gardenia jusminoidcs form glandijlora. Fragrance. 1 44-
I he orange- red tripartite stigmas from Crocus sativns flowers after drying constitute the saffron of commerce, used tor colouring and flavouring food and in the Ayurvedic system of medicine (Sastry L V L. Snruvasan M. Subrahmanyan V 1955 Saffron (Crocus xativus I..). JSci Industr Res 14A, 178-184). Saffron is valued both for its colouring and flavour properties whereas cape jasmine is a colour source only. Freshly picked saffron is virtually odourless but during drying odoriferous substances develop by degradation of bitter picrocrocm. The saffron
taste may be described as somewhat bitter, while its flavour is generally warm honey like and very pleasing. The plant C. salivus L Fam. Iridaceae), a perennial, stemless. low -growing herb with globular corms is a native of Greece and Asia-minor and now cultivated in various parts of the world from Europe to China. Today. Iran is the major supplier to the world market but its quality is not as high as that of the former supplier, Spain. India is also a major producer (approx. 30 tones per annum valued at US S20 million). Successful and significant penetration of the international market will be dependent upon introduction of quality products by India either as such in the form of saffron or through value added newer products. This patent reports a new value added product of saf&on. The total concentration of pigments (Crocins). volatile and bitter principles, in processed material was very close to that present in the corresponding starting material i.e. in saffron filaments. Keeping in yiew, its utility as a ready to serve new flavouring and colouring material, the product and process is likely to be well received by food and pharmaceutical industry. In addition, crocins obtained may also be used for anti-carcinogenic activity and other pharmacological studies [Escribano J, Alonoso G L, Salinas M R and Fernandez J A. 197. n" P970064, Spain; Garcia F E. C.A. (2000) 133: P254466b].
The chief pigments of saffron are Crocetin & its esters Crocin-1 [Crocetin-di (jS-D-gentiobiosyl) ester], Crocin-2 [CrocetinH3-D-gentiobiosyl)-(]3-D-glucosyl)-ester], Crocin-3 (Crocetm-mcno-/J-D-gentiobiosyl ester), Crocin-4 (0-D-monoglucoside ester of monomethyl a-crocetin): Crocetin-di-(/3-D-glucosyl) ester, Croceiin-mono-|8-D-ghicosylester, 13-m-Crocin along with minor amounts ofCrocetin-(j8-gentiobiosyl)-(/3-neopolitanosyl)-ester, Mangicrocin (mangiferin-6'-O-crocetyl-l"-0-0-D-glucoside ester). a-Carotene, 0-Carotene, lycopene. zeaxanthin, Neapolitanose-(( )-0-D-ghicopyranosyl-( 1 —»2)-p- Fresh stigmas were virtually odourless and most of the flavour compounds were
formed by thermal and enzymic degradation of picrocrocin [4-(8-D-glucopyranosyloxy)-2,6,6-trirrie1hyl-l-cyclohexene-l-carboxaldehyde ] into glucose, safranal (2.6.6 -trimethyl-1. 3-cyclohexadiene-l-carboxaldehyde) and 4-hydroxysafranal (2,6,6-trimedryl-4-fl-hydroxy-1-cyclobhexene-l-carboxaldehyde).Safianal further undergoes
enzymic reduction and non-en/ymic oxidation, decarboxylation and isomerization to afford isophorone-related compounds. Safianal and isophorone derivatives have been reported as the chief flavour constituents of superior grade saffron (Zarghami N S. Heinz D E 1971 Monoterpene aldehydes and isophorone related compounds of saffion. Phyiochem 102755-2761; The volatile constituents of saffion. Lehenxim Wiss Technol 42-4).
In India, saffron is produced mostly in J & K State. The conventional methods involve separation of stigma from flowers and post-harvest processing followed by drying to afford saffion of commerce. Earlier, Regional Research Laboratory, Jammu had devised a process for the quality post-harvest processing and drying of saffion to afford product of ISO Standards or even better (Raina B L, Agarwal. S G, Bhatia A K, Gaur G S 1996 Changes in Pigments and Volatiles of Saffion (Crocus sativus L.) During Processing and Storage, J Sci. FoodAgric. 71.27-32)
Most of the information on analysis and chemical composition of saffron is of academic interest. No concrete information is available in the literature for the quantitative isolation of total pigments and flavour material (ex Crocus sativus stigma or from any other source) into a value-added product economically, hence the necessity for the development of a process. The above mentioned literature survey revealed that most of the important coloring and flavour compounds of saffion are glycosidic in nature, which undergo enzymatic and thermal hydrolysis and degradation during processing and storage thus rendering it difficult to isolate quality determining coloring pigments and flavour compounds in quantitative yields Probably due to this reason, no ready to use saffion preparations are available in the market.
One of the problems faced by the food industry is that most of the carotenoid available in the market are insoluble in water and being extracted from the plant sources with organic solvents like chlorinated solvents and petroleum based aromatic hydrocarbons. Since there are restrictions on the use of solvent extracted pigments and flavors in the processed food industry, thus there is a scope for the development of a process avoiding use of organic solvents for the extraction of ready to use pigments and flavor chemicals for the food industry. The present invention provides a process, which uses so to
say natural solvents with total recovery of desirable materials in quantitative yield without solvent residues along with the separation of value added fraction of high utility from the starting materials.
Since, saffron is used from a long-long time as food additive, its harmlessness for human consumption has been more than proved, hence of the products thereof. Moreover, recently its great anti-oxidative and anti-tumor activities [Esribano. J et a). Cancer Letters! 19%) 100. 2.1-30) have been demonstrated. Saffron has a very high retail value because its cultivation and collection of flowers is a very labor-intensive process. Saffron collection in fact involves three separate manual steps consisting picking of flowers, separating of stigma and drying or post-harvest processing. To produce 1 kg of saffron, processing of nearly 1.5 lacs to 2.5 lacs flowers is necessary. Introduction of IVth processing step for the production of ready to use/serve freely water-soluble value added product may be of immense value for the food and pharmaceutical industry.
No prior art is available in the literature concerning the present invention.
Objects of the present invention
The main object of the present invention is to develop an efficient method for the
extraction of high stability, superior quality, value added, standardized, ready-to-use
saffron pigments and flavour concentrate.
Another object of the present invention is to develop a concentrate free from solvent
residues.
Yet another object of the present invention is io develop a concentrate used in
pharmaceutical, food, and allied industries.
Still another object of the present invention is to develop that helps recover very high
percentage of the saffron pigments and flavour compounds as compared to starting
material.
Still another object of the present invention is to develop saffron concentrate that does
not undergo enzymatic and thermal hydrolysis and degradation during processing.
Still another object of the present invention is to develop a water-soluble concentrate.
Still another object of the present invention is to develop a concentrate free from
decomposed impurities.
Still another object of the present invention is to develop a an odour free saffron
concentrate.
Summary of the present invention
The present invention relates to an efficient method for the extraction of high stability, superior quality, value added, standardized, ready-to-use saffron pigments and flavour concentrate, said process comprising steps of mixing saffron with one food-grade solvent or a combination of more than one food graded-solvents, macerating and agitating the mixture with continuance protection from light condition, centrifuging the macerated mixture to remove undesirable fibrous plant material, cooling the centrifuge immediately, lyophilizing the cooled centrifuge under reduced pressure to obtain crude material, isolating the concentrate by column chromatography, and obtaining brilliant orange color shining saffron pigments and flavour concentrate with recovery of about 95%, and use of the saffron pigments and flavour concentrate of range between 0.05 to 3% in food, pharmaceutical, and allied industries for flavour, and color.
Detailed description of the present invention
Accordingly, the present invention relates to a process for the isolation of saffron pigments and flavour concentrate, said process comprising steps of:
a) mixing saffron with one food-grade solvent or a combination of more than one food graded-solvents,
b) macerating and agitating the mixture with continuance protection from light condition at a temperature in the range of 5°C to 100OC,
t) centrifuging/filtering the above said macerated mixture to remove undesirable fibrous plant material,
d) cooling the centrifuge immediately to a temperature in the range of-30°C to -20°C" under men atmosphere at a pH in the range of 4 to 9,
e) lyophilizing the cooled centrifuge under reduced pressure to obtain crude material.
I") isolating the concentrate by column chromatography, and
g) obtaining brilliant orange color shining saffron pigments and flavour concentrate with recovery
of 95%. In another embodiment of the present invention, wherein the concentrate is of flavoring and pharmaceutical grade.
In yet another embodiment of the present invention, wherein the saffron is obtained from stigma, or style, or mixture of both, in filament or powder form.
In still another embodiment of the present invention, wherein concentrate is free from solvent icsidues.
In still another embodiment of the presen* invention, wherein the concentrate is free soluble in polar solvents.
In still another embodiment of the present invention, wherein the concentrate is used in pharmaceutical, food, and allied industries.
In still another embodiment of the present invention, wherein the chromatography is using silica-gel method.
In still another embodiment of the present invention, wherein said method helps recover about 95"o of the saffron pigments and flavour compounds as compared to starting material. In still another embodiment of the presen invention, wherein the pigments are crocus pigments. In still another embodiment of the present invention, wherein the saffron does not undergo enzymatic and thermal hydrolysis and degradation during processing.
In still another embodiment of the present invention, wherein the concentrate is water-soluble. In still another embodiment of the present invention, wherein the concentrate is free of solvent iesidues.
In still another embodiment of the present invention, wherein the concentrate is free from decomposed impurities.
In still another embodiment of the present invention, wherein the concentrate is odour free.
In still another embodiment of the present invention, wherein the solvents are food-grade solvents.
In still another embodiment of the present invention, wherein the extraction is at temperature is ranging between 110 to 20°C.
In still another embodiment of the present invention, wherein the atmosphere for the extraction is selected from a group comprising air, inert, hydrogen, helium, and nitrogen.
In still another embodiment of the present invention, wherein the atmosphere is protected from light conditions.
In still another embodiment of the present invention, wherein the extraction is at pH ranging between 4 to 9.
In still another embodiment of the present invention, wherein the extraction is at pH ranging between 6 to 8.
In still another embodiment of the present invention, wherein maceration is for the time duration ranging between 30 minutes and 24 hours.
In still another embodiment of the present invention, wherein the solvent is selected from a group comprising water, steam, superheated steam, alcohols, and ketones. In still another embodiment of the present invention, wherein the alcohol is of size C1 to C4, or its derivatives, and/or their mixture.
In still another embodiment of the present invention, wherein the centrifuge is cooled at the temperature ranging between 60 to ~60°C.
In still another embodiment of the present invention, wherein the centrifuge is cooled at temperature about -10OC.
In still another embodiment of the present invention, wherein rate of extraction slows down with decrease in temperature.
In still another embodiment of the present invention, wherein the rate of extraction is increased with high ratio of raw material to solvent.
In still another embodiment of the present invention, wherein increase in temperature leads to thermal decomposition of crocin pigments into undesirable crocetin.
In still another embodiment of the present invention, wherein all the parameters of the
method is critical for obtaining concentrate of desired standards.
In still another embodiment of the present invention, wherein yield of the concentrate
is ranging between 10 to 30%.
In still another embodiment of the present invention, wherein the use of the saffron
pigments and flavour concentrate in food, pharmaceutical, and allied industries for
flavour, and color.
In still another embodiment of the present invention, wherein the concentration of the
said concentrate is ranging between 0.05 to 3%.
In still another embodiment of the present invention, wherein the present invention
relates to a process for the production of a new value added product of saffron named as
Saffron pigments and flavour concentrate. The invention particularly relates to a novel and
standardized product for the production of a concentrate containing the original colour
and flavour of saffron. More particularly this in mention relates to the total extraction
of saffron pigments and flavour compounds in a single step without any enzymatic/
thermal
hydrolysis and degradation of quality determining pigments and flavoring compounds
followed by removal of solvent under suitable reduced pressure and temperature into
brilliant orange colored shining powder, in varied solvent media under optimum conditions of
temperature, pressure. pH and time to get saffron concentrate of high stable quality which
is separated from the undesirable fibrous matter by filtration/ centrifugation for the
recovery of flavoring/ pharmaceutical grade. Crocus pigments and flavour concentrate of
natural Crocus .1
In still another embodiment of the present invention, wherein accordingly, the present invention provides a new process for the production of saffron pigments and flavour concentrate which comprises direct extraction and follow up processing of saffron in a single step with food grade solvents at a temperature in the range of 110 to -10°C under natural/ inert/ Hydrogen atmosphere as such or in the dark, at a pH in the range of 4 to 9. for a period of 30 min. to 24 hr. recovering and separating the product by conventional methods.
In still another embodiment of the present invention, wherein the food grade solvents used may be such as water, steam, superheated steam, alcohols and ketones. In another embodiment of the process the atmosphere used may be that of nitrogen, air, hydrogen, helium either as such or in the dark.
In still another embodiment of the present invention, wherein thus, the process of production of saffron pigments and flavour concentrate comprises of the-direct selective extraction of filament or powder saffron in a single step with food grade solvents under continuous maceration and agitation without any enzymatic/ thermal decomposition of natural constituents, under natural/dark conditions in varied solvent media if desired, under optimum conditions of temperature in the range of 110 to -10"C. under natural/inert/Nitrogen atmosphere in natural or dark, at a pH in the range of 4 to 9 for a period of 30 min. to 24 hr. recovering and separating the product at a temperature in the range of 60 to -60° C to get Saffron pigments (colour) and flavour concentrate of high quality, which is separated from the un-extracted fibrous material by conventional methods like centrifugation or filtration.
In still another embodiment of the present invention, wherein in general, the usual relationship of time, temperature, and amount apply in the present case. Preferred temperatures range from 110 to -10° C is ideal. At lower temperature extraction tends to be slow unless a high ratio of raw material to solvent is utilized. At higher temperature, the thermal decomposition of crocin pigments to undesirable crocetin becomes an increasingly serious side reaction.
In still another embodiment of the present invention, wherein in general, the usual relationship of activity, temperature, pressure, medium and time, along with other physical and chemical parameters apply in the present case. It is important to optimize all the above-mentioned conditions in order to get the desired product.
Advantages:
1. An efficient and new one step process for the production of value added Saffron pigments and flavour concentrate of high stable quality from any type of (such as filament or powder) saffron.
2. The process provides a saffron pigments and flavour concentrate rich in natural pigments and flavoring chemicals. The total pigment concentration in the concentrate is around 10 to 25.6% depending upon the quality of starting material.
3. An improved one step process for the production of Saffron colour and flavour concentrate, which is devoid of the fibrous material and thus suitable as ready to use preparation in the food and pharmaceutical industry.
4. A new one step process for the production of Saffron pigments and flavour concentrate with typical saffron aroma and color which is freely soluble in
polar solvents including water and is free from all solvent residues. The invention is described further with reference to examples given below. The example should not be construed as to restrict the scope of the invention.
Example 1
Saffron (10g.), water (100ml.) and ethanol (100ml.) were mixed and macerated at 60°C efficiently for 4 hours in dark and centrifuged to afford an aqueous material, which was immediately cooled and subjected to lyophilization at a temperature in the range of -20° C. The crude material so obtained was passed over silica gel to afford saffron pigments and flavour concentrate (5.45g). The product was analyzed by UV spectrophotometry at 440 nm for its pigment concentration (Color value calculated as Crocin by UV estimation at 440 nm = 23%).
Example 2
Saffron (1 Ogms) and water (500ml.) were mixed and macerated at 80°C efficiently for 30 minutes and filtered to afford an aqueous material, which was stored in deep freezer. The process was repeated three times and mixed extract was concentrated at a temperature in the range of 30-40° C under inert atmosphere. The crude material so obtained was passed over alumina grade 1 to afford saffron pigments and flavour concentrate (6.73g; pigment concentration 10%).
Example 3
Saffron (l0gms), diethyl ether (200ml.) and methanol (400ml.) were mixed and macerated at 20°C efficiently for 14 hours with addition of triple distilled water (200 ml.) and centrifuged to afford an aliquot material which was subjected to rotavapor at a temperature in the range of 10"C under reduced pressure. The crude material so obtained was passed over Sephadex column to at lord saffron pigments and flavour concentrate (5.98g; pigment concentration 17.6%).
Example 4
Saffron (10g,), n-butanol (250ml.) water (100ml) and methanol (50ml.) were mixed and macerated at 5°C to -5°C efficiently for 10 hours and centrifuged to afford liquid material which was subjected to lyophilization at a temperature in the range of -30° C under reduced pressure. The crude material (6.92g) had pigment concentration of 16.6%.




We claim :
1. A process for the isolation of saffron pigments and flavour concentrate, said process
comprising steps of:
a) mixing saffron with one food-grade solvent or a combination of more than one food graded-solvents,
b) macerating and agitating the mixture with continuance protection from light condition at a temperature in the range of 5°C to 100°C,
c) centrifuging/filtering the above said macerated mixture to remove undesirable fibrous plant material,
d) cooling the centrifuge immediately to a temperature in the range of -30°C to -20°C under inert atmosphere at a pH in the range of 4 to 9,
e) lyophilizing the cooled centrifuge under reduced pressure to obtain crude material.
f) isolating the concentrate by column chromatography using silica gel method and
g) obtaining brilliant orange color shining saffron pigments and flavour concentrate with recovery of 95%.
2. A method as claimed in claim 1, wherein the saffron is obtained from stigma, or style, or
mixture of both, in filament or powder form.
A method as claimed in claims 1-2, wherein the inert gas used is for the extraction is hydrogen, helium, and nitrogen.
4. A method as claimed in claims 1-3, wherein the extraction is at pH preferably ranging between 6 to 8.
5. A method as claimed in claims 1-4, wherein maceration is for the time duration ranging between 30 minutes and 24 hours.
A method as claimed in claims 1-5, wherein the solvent is selected from a group comprising
water, steam, superheated steam, alcohols, and ketones.
A method as claimed in claims 1-6, wherein the alcohol is of size CI to C4, or its
derivatives, and/or their mixture.
A method as claimed in claims 1-7, wherein the centrifuge is cooled at the temperature
ranging between 60 to -60°C.
A method as claimed in claims 1-8, wherein the centrifuge is cooled at temperature about
10°C.
A method as claimed in claims 1-9, wherein lyophilizing the cooled centrifuge if done at a
temperature between -15 to -25°C, preferably at -20°C.
A process for the isolation of saffron pigments and flavour concentrate substantially as
herein describe with reference to examples accompanying this specification.

Documents:

1001-del-2003-abstract.pdf

1001-del-2003-claims.pdf

1001-del-2003-complete specification (granded).pdf

1001-del-2003-correspondence-others.pdf

1001-del-2003-correspondence-po.pdf

1001-del-2003-description (complete).pdf

1001-del-2003-form-1.pdf

1001-del-2003-form-13.pdf

1001-del-2003-form-19.pdf

1001-del-2003-form-2.pdf

1001-del-2003-form-3.pdf

1001-del-2003-pa.pdf


Patent Number 196771
Indian Patent Application Number 1001/DEL/2003
PG Journal Number 37/2008
Publication Date 12-Sep-2008
Grant Date 16-Mar-2007
Date of Filing 14-Aug-2003
Name of Patentee COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH, an Indian registred body incorporated under the Registration of Societies Act (Act XXI of 1860)
Applicant Address RAFI MARG, NEW DELHI-110001
Inventors:
# Inventor's Name Inventor's Address
1 S.G AGARWAL REGIONAL RESEARCH LABORATORY (CSIR),JAMMU
2 R. K . THAPPA REGIONAL RESEARCH LABORATORY (CSIR)JAMMU
3 V. K. AGNIHOTRI, REGIONAL RESEARCH LABORATORY (CSIR)JAMMU
4 O.P. SURI REGIONAL RESEARCH LABORATORY (CSIR)JAMMU
5 G. N. QAZI REGIONAL RESEARCH LABORATORY (CSIR)JAMMU
PCT International Classification Number A23L 1/221
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA