The present invention relates to a process for preparation of detoxified Simarouba glauca oilseed meal.
Simarouba glauca, commonly known as Aceituno, is a tree-borne oilseed belonging to Simaroubaceae family. Simarouba is widely grown in countries like Costa Rica, El Salvador, Honduras, Cuba, Nicaragua, Mexico, Haiti and Jamaica. Simarouba forms an important source of edible oil for various South and Central American countries. Simarouba has synonyms like Quassia simarouba, Zwingera amara, simarouba medicinalis etc. Gavilan, negrito, maruba and paradise tree are some of the common names of simarouba in different countries. Cultivation of simarouba was introduced in India as a potential source of vegetable oil during late sixties to assess its performance on marginal lands in sub-humid climate of Orissa. Later, in seventies the cultivation of simarouba spread to other Indian states like Gujarat, Maharashtra, Tamilnadu, Karnataka etc in semi arid and saline land.
Simarouba is a ever-green medium-sized tree that grows upto 15m high, with a trunk 50-80 cm. Simarouba tree produces bright green leaves 20-50 cm in length, small white flowers and small red fruits. Flowering is annual, beginning in December and continuing upto following February. Simarouba trees are polygamodioecius, with about 5% of population producing exclusively staminate flowers, 40-50% produce pistilate flowers and remaining 40-50% producing mainly staminate and few andromonoecious flowers.
Simarouba tree has an ability to grow well even in marginal waste land / dry land with degraded soils. Simarouba is a unique tree that all its parts are useful in some way or the other. Seeds are economically very important, as they contain 55-65% edible oil, which can be used in manufacture of vegetable fat or margarine. In central American countries, simarouba oil is produced and marketed for edible purposes. Simarouba oil is also used in industrial manufacture of soap, lubricant, paint, polishes, Pharmaceuticals etc. Shells (endocarp) are used in hard board industry. Fruit pulp, constituting 60% of fresh fruitlet and amounting to about 20kg / tree/ year contains 11-12% sugars, making them well suited for beverage/ fermentation industry. Leaf litter, about 20kg / tree/ year, makes
good manure. Simarouba wood, being light and insect resistant, is useful in furniture making. The bark and leaf of simarouba contain simarubin, a triterpene useful in curing amoebiasis, diarrhoea and malaria. The oil cake of simarouba rich in nitrogen (7.5-8.0%), phosphorus (1%) and potash (1.5%) is valuable organic manure.
The seeds of simarouba contain about 30-35% kernels and 65-70% shells. Simarouba kernels are very rich source of oil with its content varying between 55 and 65%. Simarouba oil is being used for edible as well as industrial purposes. Protein content in simarouba kernel ranges between 15 and 20%. Residual meal cake of simarouba, after its oil extraction, contains high protein (45-55%). However, the meal as well as kernel has persistent bitter taste. Presence of toxic quassine and other anti nutritional factors render simarouba meal unfit for food / feed purposes in spite of presence of high protein of good quality. Main toxic factors of simarouba have been reported to belong to triterpene family. As the residual meal cake obtained from simarouba, which is toxic to livestock, is disposed-off as manure (Mario Lewy-Van Severen. 1953 March. Journal of Am. Oil Chemist Society. 124-126 pp., Rao, KVSA and Lakshminarayana, G. 1983 July/Aug. J. Fd. Sci. Tech. 176-177 pp., Chikara J, Shethia BD, Meena Rathod and Pandya, JB. 1998 Oct/Dec. J. Oil Technologists Association of India 30(4) 177-179 pp).
Literature pertaining to simarouba deals mainly with therapeutic properties of its quassinoids. Patent search clearly indicates that no patent deals with detoxification process of simarouba. Among the patents pertaining to therapeutic properties, Grieco, PA, 2003 US Patent # 6573296) have reported antineoplastic, antiviral and herbistatic activity in quassinoid preparations. Simarouba is the subject of one US patent so far, whereby its water extract was found to have cosmetic or pharmaceutical property, having significant depigmentation activity with ability to enhance the protective water barrier function of skin and keratinocyte differentiation (Bonte, F. 1997. US Patent # 5676948).
Population in South American countries like Cuba, Brazil, Guyana have used simarouba bark extract for fever, malaria, dysentery, as hemostatic agent to stop bleeding and as tonic.
The Merck Institute reported that simarouba was 91.8% effective against intestinal amebas in humans. Scientists have again documented the simarouba's ability to kill common dysentery-causing organism, Entamoeba histolytica, Salmonella and Shigella. Studies have also demonstrated that simarouba and/or its potent quassinoids were effective against malaria in vitro as well as in vivo (O'Neill, MJ. et al. 1988. J. Ethnopharmacol 22.2. 183-190 pp).
Although there is no scientific report available pertaining to detoxification of simarouba meal, few earlier reports deal with detoxification of meal of other tree-borne oilseeds.
Reference may be made to the publication of Aregheore, EM, Becker, K and Makkar, HPS (S. Pacific J. Nat. Sci. 2003. 21. 50-56 pp) whereby heat and 14 different chemical treatments have been used to detoxify the meal of Jatropha curcas from lectin and phorbol esters. Chemical detoxification of Jatropha included treatment with sodium hydroxide and sodium hypochlorite. The main draw back of this reference is that the treatments though inactivated lectin but not phorbol ester of Jatropha. Further, treatments have not been applicable to Simarouba, where in major toxic and antinutritional factors are saponins and alkaloids.
Reference may also be made to the publication of Makkar, HPS, Becker, K and Schmook, B (Plant Foods for Human Nutrition), whereby roasting has been employed to detoxify Jatropha curcas. Report indicates that roasting inactivated 100% trypsin inhibitor, but lectin was reduced partly. The main draw back of this reference is that the treatment has not been applicable to Simarouba. Further, report also indicates that roasting has not affected the saponins, phytate and phorbol esters of Jatropha.
Reference may also be made to the publication of Aderibigbe, AO, Johnson, COLE, Makkar, HPS, Becker, K and Foidl, M (Animal Feed Science Technology 1997. 67. 223-243 pp), whereby several dry and wet heat treatments have been used to detoxify Jatropha meal, whereby trypsin inhibitor activity has been reduced. However, the main draw back of the report is that the treatments have not been applicable to Simarouba.
Further, the used heat treatments could not decrease the antinutritional factors like saponin, lectins and phytate.
Reference may also be made to the publication of Shanmugasundaram, T and Venkataraman, LV (J. Sci. Food Agric. 1985. 36. 1189-1192 pp), wherein 20, 40, 60, 80 and 95% ethanol extractions have been used to detoxify Medhuca seed flour from saponin. The main draw back of the report is that the treatments have not been applicable to Simarouba. Further, effect of ethanol extraction on removal of alkaloid has not been indicated.
Reference may be made to the publication of Barrett JE, Klopfenstein CF and Leipold HW (Plant Foods for Human Nutrition. 1998. 52. 9-15 pp), wherein canola and rapeseed meals are treated with alkali (5% NaHCO3 and 5% NH4HCO3) and heated to partly remove glucosinolates. The main draw back of the report is that the treatments have not been applicable to Simarouba. Further, influence of alkali treatment on antinutritional factors like saponins and alkaloids are not indicated.
Reference may be made to publication of Klans Dieter Schwenke, Jurgen Kroll, Reinhard Lange, Mantred Kujawa and Werner Schnaak (J. Sci. Food Agric. 1990. 51. 391-405 pp), wherein mild soaking in citric acid or ammonium carbonate is used to detoxify rapeseed flour from glucosinolates. The main draw back of the report pertains to non-applicability of treatment to Simarouba, whereby toxic constituents are different as saponins and alkaloids.
Reference may be made to publication of Behura NC, Dehuri PK, Mishra SK and Samal PC (Animal Nutrition and Feed Technology, 2008. 8. 245-252) wherein different physico-chemical treatments are used to detoxify Simarouba oilcake. However, main drawback of the process pertains to loss of protein content in detoxified Simarouba meal, complete lack of quantification of toxic constituents to claim detoxification of Simarouba, objectively.
The main objective of the present invention is to detoxify simarouba meal.
Another objective of the present invention is to detoxify simarouba meal by reducing saponin.
Yet another objective of the present invention is to detoxify simarouba meal by reducing alkaloid
Still another objective of the present invention is to prepare simarouba meal rich in protein content
Yet another objective of the present invention is to evaluate the safety and detoxification efficacy in rat growth study and Protein Efficiency Ratio (PER) assessment.
Accordingly, the present invention provides the process for detoxification of simarouba meal, which comprises following steps:
(a) grinding Simarouba kernels into coarse particles followed by extraction in hexane
solvent to defat the Simarouba kernel and to obtain the meal with less than 1.0%
fat,
(b) boiling defatted Simarouba meal in organic solvent selected from 2% acetic acid
in ratio 1:10 (w/v) optionally extraction with mixture of ethanol and 2% acetic
acid (1:8) for 30 min at temperature range of 95-98° C at pH 3.8-4.4,
(c) filtering the residue of Simarouba meal and washing with 2% acetic acid solution,
(d) re-boiling the filtered and washed Simarouba meal in 2% acetic acid in the ratio
of 1:10 to 1:12 for 30 min at temperature range of 95-98° C at pH 3.5-4.0,
(e) filtering the obtained residue of Simarouba meal of step (d) and drying in hot air
oven at 60° C to attain the moisture content of 6-7%,
(f) powdering the dried meal of step (e) to get the desired detoxified Simarouba meal.
In an embodiment of the present invention, following critical parameters are identified:
1. Acidic pH of detoxifying medium (3.5-4.4 pH)
2. Minimum of two boiling treatments, in acetic acid medium
3. Temperature of acid extraction (95-98°C)- boiling condition
In an embodiment of the present invention, decorticated simarouba kernels were ground into coarse particles. Six extractions in Soxhlet unit using hexane solvent were employed to defat the simarouba kernel particles and to obtain the meal with 1-1.5% fat content.
In another embodiment of the present invention defatted simarouba meal (lOOg) was taken in 2% acetic acid in ratio 1:10 (w/v) and subjected to boiling for 30 min.
In yet another embodiment of the present invention, residue simarouba meal after boiling was filtered-off, decanting the medium. Filtered simarouba meal was washed with 2% acetic acid solution.
In yet another embodiment of the present invention, filtered and washed simarouba meal, after its first boiling was subjected to boiling second time in 2% acetic acid in the ratio of 1:12 for 30 min.
hi still another embodiment of the present invention, residue simarouba meal after boiling second time, was filtered-off, decanting the medium. Simarouba meal thus boiled and filtered was dried in hot air oven at 60°C to attain the moisture content of 6-7%. Thus treated and detoxified simarouba meal was powdered.
In yet another embodiment of the present invention,detoxified simarouba meal as obtained at step (f) contained negligible levels of toxicants like saponin (-0.15%) and alkaloid (-0.09%) and higher protein content (-60%) and in-vitro-protein digestibility (84-86%).
In still another embodiment of the present invention the detoxified simarouba meal prepared, is a fine powder of free-flow nature, having creamish brown color with no off-taste.
In another embodiment of present invention, the efficacy of thus detoxified Simarouba meal was evaluated for safety in rat growth study and PER.
The detoxification treatment used in the invention resulted in following properties in Simarouba meal:
i. Reduction in saponin level of simarouba meal:
Untreated simarouba meal contained saponins at the level of 3.70%. Detoxification process as mentioned at steps a-e caused reduction in saponin content upto the extent of 94% and detoxified meal contained negligible level of saponin (0.15%). ii. Reduction in alkaloid level of simarouba meal:
Untreated simarouba meal contained alkaloid at the level of 1.0%. Detoxification process, as mentioned at steps a-e caused reduction in alkaloid content upto the extent of 91% and detoxified meal contained negligible level of alkaloid (0.09%). Hi. Increase in protein content of simarouba meal:
Detoxification process, as mentioned at step a-e caused increase in protein content of simarouba meal by over 27%. Untreated simarouba meal contained protein at the level of 47.7% and the protein content increased to 60.7% in detoxified simarouba meal, iv. Safety evaluation of detoxified Simarouba meal in rat growth study:
Efficacy of detoxification process, as mentioned at step a-f determined in rat growth study, following standard method (IS 7481-1974) indicated the PER of 1.90.
Evaluation of efficacy of detoxification process of Simarouba meal, as followed in present invention indicated PER of 1.90 in rat bio-assay growth study
(Table Removed) Novelty of Invention:
1. Simple, adoptable process using 2% acetic acid under double boiling
condition, which simultaneously removes saponins and alkaloids by 91-
94%.
2. Simarouba meal thus detoxified had higher protein content (-60%) with
high in-vitro protein digestibility.
3. Detoxified Simarouba meal thus obtained with Protein Efficiency Ratio
(PER) of 1.90 for 50% replacement of protein source, has potential for use
in animal feed formulations.
The flow chart of process for detoxification of Simarouba meal:
(Formula Removed)
The following examples are given by way of illustration of the present invention and therefore should not be construed to limit the scope of present invention. Variation and changes may be made by one skilled in art without departing from spirit of the invention. All parts and percentages are by weight, unless otherwise indicated.
Example 1
In the present examples, untreated and reference simarouba meal was prepared without any detoxification treatment. Decorticated simarouba seeds were broken into bits, using Roller Flaker. Six extractions in Soxhlet unit using hexane solvent were employed to defat the simarouba kernel particles and to obtain the meal with 1-1.5% fat content. Untreated simarouba meal without any detoxification treatment contained following constituents.
(Table Removed)
Example 2
Defatted simarouba meal (lOOg) was dispersed in a ratio of 1:20 (w/v, pH 6.8) and boiled (95-98°C) for 30 min. The slurry was filtered and washed with water. The residue was dried in a hot air oven at 60°C to a moisture level of 6-7%. Detoxified simarouba meal obtained by above process contained following constituents.
(Table Removed) Example 3
Defatted simarouba meal (lOOg) was taken in 2% acetic acid solution in the ratio 1:10 (w/v, pH 3.8-4.4) and autoclaved at 1 kg/cm2 for 30 min. Simarouba meal residue thus obtained was washed again with 2% acetic acid. Resulting meal was dried in a hot air oven at 60°C to a moisture level of 6-7%. Detoxified simarouba meal thus obtained contained following constituents.
Example 4
Defatted simarouba meal (l00g) was taken in 2% acetic acid solution in the ratio 1:10 (w/v, pH 3.8-4.4) and boiled (95-98°C) for 30 min and filtered. The residue meal was washed with 2% acetic acid and dried in a hot air oven at 60°C to a moisture level of 6-7%. Thus detoxified and powdered simarouba meal contained following constituents.
(Table Removed)
Example 5
Known weight of defatted simarouba meal (lOOg) was dispersed in 1% NaOH in the ratio 1:10 (w/v) into a conical flask and stirred for 1 hour on a rotary shaker. The residue meal after extraction was filtered and washed with 1% NaOH. The simarouba meal thus detoxified was dried in a hot air oven at 60°C to a moisture level of 6-7%. Dried and powdered detoxified simarouba meal contained following constituents.
(Table Removed)
Example 6
Defatted simarouba meal (lOOg) was taken in 70% ethanol (1:8; w/v) containing 2% acetic acid (v/v, pH 3.8-4.4). The contents in conical flask were stirred for 1 hour on a Rotary Shaker. The residue meal after extraction was filtered and dried in hot air oven at 60°C to a moisture content of 6-7%. Dried and powdered detoxified simarouba meal contained following constituents.
(Table Removed)
Example 7
A known weight of simarouba meal (l00g) was taken in 2% acetic acid at the ratio 1:10 (w/v, pH 3.8-4.4) and cold extracted for 2 hrs on Rotary Shaker. The meal after extraction was filtered off and dried in a hot air oven at 60°C to moisture content of 6-7%. Treated and dried simarouba meal was powdered. The simarouba meal thus detoxified contained following constituents.
(Table Removed)
Example 8
A known weight of defatted simarouba meal (lOOg) was packed in a wide percolating column and saturated with 2% acetic acid. Meal was left to remain in contact with 2% acetic acid solution in column for 6 hrs and then drained. Similar sequential percolation of meal was done with 2% acetic acid for four times. After four percolations, meal was squeezed and kept in oven at 60°C till moisture content of 6-7%. Simarouba meal thus detoxified contained following constituents.
(Table Removed)
Example 9
Defatted simarouba meal (lOOg) was taken in 2% acetic acid at the ratio 1:6 (w/v, pH 3.8-4.4) and kept at rotary shaker for 6 hrs. The same treatment was followed for four extractions and each extraction was followed by pressing in nylon cloth. At the end of fourth extraction, meal was dried at 60°C till the moisture content was 6-7%. This detoxified simarouba meal contained the following constituents.
(Table Removed) Example 10
Defatted simarouba meal (lOOg) was taken in 2% acetic acid at the ratio 1:10 (w/v, 3.8-4.4 pH) and subjected to boiling ((95-98°C) for 30 min. Residue simarouba meal was filtered off, decanting the acid medium. Filtered simarouba meal was washed with 2% acetic acid solution. Simarouba meal after its first boiling was subjected to second boiling (95-98°C) in 2% acetic acid in the ratio 1:10 (w/v, pH 3.5-4.0) for 30 min. Simarouba meal was filtered decanting acid medium. Simarouba meal thus treated was dried in hot air oven at 60°C to attain moisture content of 6-7%. Detoxified simarouba meal was powdered and meal thus obtained contained following constituents.
(Table Removed) Advantages of the invention,
Detoxified simarouba meal thus prepared, employing defatting, two-stage boiling in acid, washing and drying, has the following advantages.
a) Process resulting in detoxified simarouba meal contained negligible level of
saponin, which was reduced by 94%, when compared to its original content.
(Table Removed) b) Process resulting in detoxified simarouba meal contained negligible level of
alkaloids, which was reduced by 91%, when compared to its original content.
(Table Removed)
c) Process resulting in detoxified simarouba meal also contained higher protein
content. Protein content in detoxified simarouba meal increased by over 27%.

We claim:
1. A process for preparation of detoxified simarouba glauca oilseed meal which comprises the steps of:
a) grinding Simarouba kernels into coarse particles followed by extraction in
hexane solvent to defat the Simarouba kernel and to obtain the meal with less
than 1.0% fat,
b) boiling defatted Simarouba meal in organic solvent selected from 2%
acetic acid in ratio 1:10 (w/v), optionally extracting with a mixture of ethanol
and 2% acetic acid (1:8), for 30 min at temperature range of 95-98° C at pH
3.8-4.4,
c) filtering the residue of Simarouba meal and washing with 2% acetic acid
solution,
d) re-boiling the filtered and washed Simarouba meal in 2% acetic acid in the
ratio of 1:10 to 1:12 for 30 min at temperature range of 95-98° C at pH 3.5-4.0,
e) filtering the obtained residue of Simarouba meal of step (d) and drying in
hot air oven at 60° C to attain the moisture content of 6-7%,
f) powdering the dried meal of step (e) to get the desired detoxified
Simarouba meal.
2. A process, as claimed in claim 1, wherein detoxified simarouba meal as obtained
at step (f) contained negligible levels of toxicants like saponin (-0.15%) and alkaloid (~0.09%) and higher protein content (~60%) and in-vitro-protein digestibility (84-86%).
3. A process, as claimed in claim 1-2, wherein detoxified simarouba meal prepared,
is a fine powder of free-flow nature, having creamish brown color with no off-
taste.
4. A process, as claimed in claim 1-3 , wherein detoxified simarouba meal as
obtained at step (h) is useful as ingredient in animal feed.
5. A process for preparation of detoxified Simarouba glauca oilseed meal substantially as herein described with reference no. 6 and 10 .