Title of Invention

"A PROCESS FOR THE PREPARATION OF BIXIN DYE FROM ANNATTO (BIXA ORELLANA L) SEEDS"

Abstract The present invention relates to a process for the preparation of bixin dye from annatto (Bixa orellana L) seeds. Annatto extracts obtained from the seeds of the tropical shrub Bixa orellana L.have wide spread use in the food industry for the colouring of many commodities. The principal colouring component of annatto seeds is the liposoluble diapocarotenoid 9'-cis-bixin, which is the monomethyl ester of the water soluble 9'-cis-norbixin. The process involves the removal of unwanted resinous matter using hexane/petroleum ether from annatto seeds by adopting soxhlet extractions techniques for about 8-12h. The same seeds are again subjected to soxhlet extraction by using acetone for an 8-12h. The dye settled in the recovery flask during the soxhlet extraction is collected as high bixin dye (66-92% bixin) by separating the miscella using techniques such as decantation/ siphoning/ filtration/ centrifugation
Full Text The present invention relates to a process for the preparation of bixin dye from annatto (Bixa orellana L) seeds.
Annatto extracts obtained from the seeds of the tropical shrub Bixa orellana Lhave wide spread use in the food industry for the colouring of many commodities. The principal colouring component of annatto seeds is the liposoluble diapocarotenoid 9'-cis-bixin, which is the monomethyl ester of the water soluble 9'-cis-norbixin. Annatto extracts are usually formulated to provide suitably stabilized forms in the colour range red-orange-yellow for use in a wide range of foods such as dairy products, flour confectionary, fish, soft drinks, meat products, snack foods, and dry mixes.
The bixin content of the seed varies widely depending upon the agro climatic conditions and agricultural practices. The bixin content of local varieties varies between 0.7-2.0 %. Extraction of the coloring component is by different methods. General practices are:
- boiling with vegetable oil,
- boiling with alkali,
- Organic solvents.
Reference may be made to three main commercial process which are used to extract the pigment from dried annatto seeds (H.D.Preston and M.D.Rickard, Food chemistry, 5, 47-56, 1980) where in (1) direct extraction into oil, (2) direct extraction into aqueous alkali, or (3) indirect extraction of annatto pigment into suitable solvent such as acetone were discussed and the extracts being washed with hexane, or some other solvent in which the pigment was virtually insoluble to remove impurities and odours.
Reference may be made to various constituents isolated and identified from Bixa orellana (Anil srivastava, Y.N.Shukla, S.P.Jain and Sushil Kumar, Journal of Medicinal and Aromatic Plant Sciences 21, 1145-1154, 1999) wherein bixin is the oil soluble carotenoid principle colouring matter present in the seeds and is accompanied by small amout of orellin, a water soluble yellow substances,
fat, resin and a bitter stuff. Alkaline hydrolysis of bixin yielded more stable and more soluble trans bixin.
Reference may be made to preparation of annatto dyes from Bixa orellana seeds (G.V. Krishna murthy and N. Giridhar, Proceedings of the first Indian Convention of Food Scientists and Technologists, P.78, 1979) where in a method was described for obtaining a crude annatto preparation, soluble in water, alcohol or organic solvents that could be easily incorporated into foods. The seeds were extracted with 0.2% Na2C03 solutions and precipitated with acid to yield 9.3% by wt. of total dye. The dark red powder had an acid-soluble and an water-soluble component, and was soluble to varying extents in ethanol, benzene and butanol. The water-soluble dye slowly separated out in orange squash and synthetic syrup but was stable for two months in totapuri mango pulp and tomato ketchup. The colour was stable when added to solution or suspension with pH greater than 5.0.
Reference may be made to methods of extracting total pigments from annatto seeds (SV Bhalker and PJ Dubash, Indian Journal of Dairy Sciences, 36(2), 157-167, 1983) wherein water, sodium carbonate, NaOH or solvent extraction (with 5, 10, 15 and 20ml of 25% chloroform in ethanol v/v) were investigated. The various methods were critically evaluated and compared maximum extraction rate percent of total pigment was : water dislodging, 60-65; alkali extraction with NaOH, 73.9; and solvent extraction 76.48. Optimum concentration of Na2C03 was 0.4% and a maximum extraction of pigments was achieved after about 5h shaking. Water dislodging was quite time consuming (6h shaking) and required many unit operations. 74% was extracted inlh by the NaOH method, but ultimately the dye had to be dissolved in the solvent. Solvent extraction gave a maximum of 76% total pigment. Solvent extraction was therefore found to be the simplest and the best method for extracting annatto pigments.
Reference may be made to the process on "Production of crystalline dye from Annatto" developed by CFTRI, Mysore, wherein the yield of dye has been mentioned as 3% with 30-35% bixin and very rarely dye containing 50% bixin
depending on the quality of the raw material. The process is based on extraction of the seed in a battery of percolators at room temperature using organic solvents.
Patent search on the annatto reveals that the reports are mainly pertaining to preparation of different formulations and applications in various types of foods. The patent search also reveals that there is no work reported on the production of high bixin dye, which is an intermediate for different types of annatto dye formulations to suit to the requirements of various sectors of food products.
The main object of the present invention is to provide a process for the preparation of bixin dye from annatto (Bixa orellana L) seeds which is essential for production of different types of annatto dye formulations to suit to the colour requirements of a wide range of foods.
Another object of present invention is to produce high bixin content dye from annatto seeds to make the process more techno-economical and viable.
Accordingly the present invention provides a process a process for the preparation of bixin dye from annatto (Bixa orellana L) seeds, comprising;
a) extracting bixin from annatto seeds using solvents selected from hexane, and petroleum ether in the ratio of 1:5,
b) extracting crude pigment using acetone,
c) separating sediment from the above obtained crude pigment,
d) recovering solvents from miscella to obtain low bixin dye,
e) drying moist dyes in vacuum oven to recover bixin dye

In an another embodiment of the process the solvent used for extraction is in the range of 2-2.5 times the weight of annatto seed.
In an another embodiment of the process the solvent used for extraction is from a group of hexane, petroleum ether and acetone.
The process of the present invention may be carried out as follows:
The process involves the removal of unwanted resinous matter using hexane/petroleum ether from annatto seeds by adopting soxhiet extractions techniques for about 8-12h. The same seeds are again subjected to soxhiet extraction by using acetone for an 8-12h. The dye settled in the recovery flask during the soxhiet extraction is collected as high bixin dye (66-92% bixin) by separating the miscella using techniques such as decantation/ siphoning/ filtration/ centrifugation. The solvent is recovered from miscella by distillation. The dye was kept under vacuum drying at 40-50C by applying 20-25mm Hg vacuum for 2-4h to remove the traces of solvent. The yield of dye was about 0.55-1.0%.
The following examples are given by way of illustration of the present invention and therefore should not be construed to limit the scope of the present invention.
Example-1
The annatto seed (400g) was loaded in to the middle piece of a soxhiet extraction system and extracted with 2 liters of solvent hexane (LR grade, Sd fine Chemicals.Mumbai). The extraction is carried out for a period of 8 hours. The solvent is recovered to separate the resinous matter and this solvent can be reused. The seed in the soxhiet middle piece is further extracted with 2 litres of solvent acetone (LR grade, Sd fine Chemicals.Mumbai) in a soxhiet extraction system for a period of 10 hours. The crystalline sediment which is clearly visible at the bottom of collection flask is separated from the miscella to obtain high bixin
dye (4.12 g with 65.8% bixin) by decanting. After separation the miscella is distilled to recover the solvent acetone for reuse.
Example-2
The annatto seed (450g) was loaded in to the middle piece of a soxhiet extraction system and extracted with 2 liters of solvent hexane (LR grade, Sd fine Chemicals.Mumbai). The extraction is carried out for a period of 12 hours. The solvent is recovered to separate the resinous matter and this solvent can be reused. The seed in the soxhiet middle piece was further extracted with 2 liters of solvent acetone (LR grade, Sd fine Chemicals.Mumbai) in a soxhiet extraction system for a period of 12 hours. The crystalline sediment, which is clearly visible at the bottom of collection flask is separated from the miscella to obtain high bixin dye (4.78 g with 91.5% bixin) by decanting. After separation the miscella was distilled to recover the solvent acetone for reuse.
Example-3
The annatto seed (400g) was loaded in to the middle piece of a soxhiet extraction system and extracted with 2 litres of solvent hexane (LR grade, Sd fine Chemicals.Mumbai). The extraction is carried out for a period of 8 hours. The solvent is recovered to separate the resinous matter and this solvent can be reused. The seed in the soxhiet middle piece was further extracted with 2 liters of solvent acetone (LR grade, Sd fine Chemicals, Mumbai) in a soxhiet extraction system for a period of 8 hours. The crystalline sediment, which is clearly visible at the bottom of collection flask is separated from the miscella to obtain high bixin dye (2.21 g with 81.5% bixin) by decanting. After separation the miscella is distilled to recover the solvent for reuse.
The main advantages of the present invention are :
1. The bixin dye plays a major role in production of different annatto dye
formulations.
2. To produce stable annatto dye formulations with higher percent of bixin. especially oil soluble and oil soluble emulsions, high bixin annatto dye is required.
3. The dye containing high bixin has a better potential in the export market.
4. This method improves the process economics.





We claim:
1. A process for the preparation of bixin dye from annatto (Bixa orellana L) seeds,
comprising:
a) extracting bixin from annatto seeds using solvents selected from hexane, and petroleum ether in the ratio of 1:5,
b) extracting crude pigment using acetone,
c) separating sediment from the above obtained crude pigment,
d) recovering solvents from miscella to obtain low bixin dye,
e) drying moist dyes in vacuum oven to recover bixin dye

2. A process as claimed in claim 1, wherein the solvent used for extraction is in the range of 2-2.5 times the weight of annatto seed.
3. A process for the preparation of bixin dye from annatto {Bixa orellana L) seeds substantially as herein described with reference to the examples.

Documents:

552-del-2004-Abstract (27-11-2009).pdf

552-del-2004-abstract.pdf

552-del-2004-Claims (27-11-2009).pdf

552-del-2004-claims.pdf

552-del-2004-Correspondence-Others (27-11-2009).pdf

552-del-2004-Correspondence-Others-(08-04-2010).pdf

552-del-2004-correspondence-others.pdf

552-del-2004-correspondence-po.pdf

552-del-2004-Description (Complete) (27-11-2009).pdf

552-del-2004-description (complete).pdf

552-del-2004-form-1.pdf

552-del-2004-form-18.pdf

552-del-2004-form-2.pdf

552-del-2004-Form-3 (27-11-2009).pdf

552-del-2004-form-3.pdf

552-del-2004-form-5.pdf


Patent Number 244511
Indian Patent Application Number 552/DEL/2004
PG Journal Number 50/2010
Publication Date 10-Dec-2010
Grant Date 09-Dec-2010
Date of Filing 22-Mar-2004
Name of Patentee COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH
Applicant Address RAFI MARG, NEW DELHI-110001, INDIA.
Inventors:
# Inventor's Name Inventor's Address
1 PAMIDIGHANTAM PRABHAKARA RAO CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE-570 013, INDIA.
2 GALLA NARSING RAO CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE-570 013, INDIA.
3 AKULA SATYANARAYANA CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE-570 013, INDIA.
4 DUBASI GOVARDHAN RAO CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, MYSORE-570 013, INDIA.
PCT International Classification Number C 09 B 61/00
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA