ES2542163B1 - PROCEDURE FOR OBTAINING GERMINATED INTEGRAL RICE GRAINS AND PRODUCTS DERIVED FROM THEMSELVES THAT INCLUDE BIOLOGICALLY ACTIVE COMPOUNDS - Google Patents

Abstract

Process for obtaining germinated brown rice grains and products derived therefrom that comprise biologically active compounds. # The present invention relates to a simple and economical germination process that, using light, allows to obtain germinated brown rice grains and / or products derived therefrom with high contents of biologically active compounds, preferably {ga} -orizanol and GABA. The germinated brown rice grains obtained or the products derived from them are used in the preparation of food or cosmetic compositions, which in addition to their nutritional value, comprise a high content of healthy bioactive compounds, which make them recommended for both a complete diet , as for the prevention and / or reduction of the risk of the appearance of chronic degenerative pathologies.

Description

PROCEDURE FOR OBTAINING GERMAN INTEGRAL RICE GRAINS AND PRODUCTS DERIVED FROM THEMSELVES THAT INCLUDE BIOLOGICALLY ACTIVE COMPOUNDS

DESCRIPTION 5

SECTOR AND OBJECT OF THE INVENTION

The present invention relates to the agri-food sector since it specifically refers to both a method of obtaining germinated brown rice grains with 10 high contents of biologically active compounds, such as germinated brown rice grains or products derived from they are obtained from said process, such as the use thereof to obtain healthy and / or cosmetic food compositions, and directly obtained compositions.

 fifteen

STATE OF THE TECHNIQUE

Germination improves the texture and organoleptic properties of brown rice grains and nutrients are more easily digested and absorbed. The current interest aroused in sprouted brown rice is mainly due to the positive effect of the germination process on certain bioactive compounds that play an important role in the prevention of chronic diseases, among which are -oryzanol and acid gamma-aminobutyric acid (GABA).

Or-oryzanol is a potent antioxidant of relevant pharmacological, nutritional and cosmetic interest for its antioxidant, hypolipidemic effects and its ability to reduce the risk of cardiovascular diseases (Ghatak and Pamchal. Gamma-oryzanol - A multi-purpose steryl ferulate. Curr Nutr & Food Sci; 2011; 7: 10-20), for alleviating menopausal symptoms (Ishihara et al. Clinical effect of gamma-oryzanol on climacteric disturbance on serum lipid peroxides.
Nihon Sanka Fujinka Gakkai Zasshi; 1982; 34: 243-51), and as an antidiabetic agent (Imam et al. Antidiabetic properties of germinated Brown rice: a systematic review, Evidence-based Complementary and Alternative Medicine, 2012; DOI: 10.1155 / 2012/816501), to which also anti-cancer properties have been associated (Hirose et al. Modifying effects of phytic acid and gamma-oryzanol on the promotion stage of rat carcinogenesis.
Anticancer Res; 1999; 9: 3665-70). 35

For its part, GABA is a potent neurotransmitter and exerts beneficial health effects such as the regulation of blood pressure and heart rate, stimulates insulin secretion contributing to the prevention of diabetes, relieves pain, anxiety and insomnia (Patil and Khan. Germinated Brown rice as a value added rice product: A review. J. Food Sci. Technol; 2011: 48: 661-667). He has been assigned an important role in learning and memory and has been associated with reducing the risk of Alzheimer's disease (Mamiya et al. Effects of pre-germinated Brown rice on depression-like behavior in mice. Pharmacol. Biochem. Behav; 2007: 86: 62-67).

For all the above, there is a growing interest in obtaining food and / or 10 food or cosmetic preparations enriched in -oryzanol and GABA and the scientific literature shows that sprouted brown rice is an excellent natural source of these compounds, easily prepared and of attractive consumption (Wu et al. Germinated brown rice and its role in human health. Cri. Rev. Food Sci. Nutr; 2013: 53: 451-463).

 fifteen

The scientific references concerning the variations in the content of or-oryzanol during the germination of brown rice are not coincidental. This seems mainly due to the differences between varieties and between the main germination parameters, namely time, temperature and quantity of water used during the process, being jobs in which the content of this compound increases, is maintained and even decreases as 20 consequence of the process (Moongngarm and Saetung. Comparison of chemical compositions and bioactive compounds of germinated rice and Brown rice. Food Chem; 2010; 122: 782-88; Jayadeep and Malleshi. Nutrients, composition of tocopherols, tocotrienols, and -oryzanol , and antioxidant activity in brown rice before and after biotransformation. CyTA. Journal of Food. 2011; 9: 82-87; Kim et al. Chemical and functional components in different parts of rough rice 25 (Oryza sativa L.) before and after germination Food Chem; 2012; 134: 288-293). In contrast, the data found in the literature to date on the effect of germination on the GABA content of brown rice are quite coincidental and indicate that this compound generally increases with germination time (Cáceres et al. Maximizing the phytochemical content and antioxidant activity of Ecuadorian brown rice 30 sprouts through optimal germination conditions. Food Chem; DOI: 10.1016 / j. foodchem. 2013.11.156.).

The germination of rice begins with the hydration of the seed, at which time the metabolism reactivation and the hydrolysis of proteins and carbohydrates occur.

reserve, while synthesizing and accumulating new metabolites, some of them beneficial to health. The content of these bioactive compounds, therefore, will depend on the germination conditions.

EXPLANATION OF THE INVENTION 5

The first object of the invention relates to a process for obtaining brown rice grains and / or products derived therefrom that comprise biologically active compounds, characterized in that it comprises:

 10

a) in the presence of light, germinate brown rice grains for a period of time between 20 and 120 h and at a temperature between 25 and 34 ° C.

Biologically active compounds are chosen from compounds with antioxidant activity and / or compounds with antihypertensive and / or antidiabetic and / or antidepressant activity, which are preferably or-oryzanol and gamma-aminobutyric acid (GABA).

In a particular embodiment, in step a) the time is 24 hours, the temperature is 28 ° C and the or-oryzanol comprises a content greater than 14 mg per 100 g of dried flour of germinated brown rice grains and a content of GABA greater than 23 mg per 100 g of dried flour from germinated brown rice grains. twenty

The second object of the invention relates to the product obtained by the process, preferably, germinated brown rice grains or a flour obtained from the rice grains.

 25

The third object of the invention relates to the use of the product obtained by the process, in the preparation of healthy food compositions.

The fourth object of the invention relates to the use of the product obtained by the process, in the preparation of cosmetic compositions. 30

The fifth object of the invention relates to healthy food compositions and / or directly obtained cosmetic compositions.

DETAILED DESCRIPTION OF THE INVENTION 35

The technical problem solved by the present invention is to obtain germinated brown rice grains and / or products derived therefrom that have high contents of biologically active compounds, notably increased with respect to the content of these same compounds in brown rice grains. obtained by the procedures currently known in the state of the art. 5

The inventors of the present invention application have discovered that the presence of light during the germination of brown rice grains is a germination parameter that, surprisingly, allows to obtain germinated brown rice grains with increased contents of -oryzanol and GABA, which they are markedly superior to those of the same 10 germinated rice grains in the absence of light and to those of the same non-germinated grains (see Examples 3 and 4).

The main technical advantages of the invention presented here are:

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a) with respect to the procedure:

- it is economical, simple and environmentally friendly,

-

b) with respect to the product obtained by the procedure: in the case of 20 germinated rice grains, these can be easily included in the diet of consumers either directly in their fresh natural state, or transformed into flours for incorporation into the formulation of new foods or in the form of instant prepared foods.

As indicated in the prior art section, it is known to use germination procedures to increase the content of biologically active compounds. However, and in the knowledge of the inventors of the present patent application, among the parameters involved in germination, to date the use of light during the germination of brown rice seeds had not been considered to increase the content of biologically active compounds. 30

It is the first object of the invention, a process for obtaining brown rice grains and / or products derived therefrom that comprise biologically active compounds, hereinafter method of obtaining the invention, comprising:

a) in the presence of light, germinate brown rice grains for a period of time between 20 and 120 h, at a temperature between 25 and 34 ° C.

In the present invention, "biologically active compounds" means any substance effective for the maintenance of health or for reducing the risk of suffering from 5 diseases.

In one aspect of the invention, the biologically active compound is an antioxidant, and is preferably -oryzanol.

 10

By "-oryzanol" is meant the mixture of esters of triterpenic alcohols of ferulic acid found mostly in the integral rice husk, and which have been associated with antioxidant properties beneficial to health.

In another aspect of the invention, the biologically active compound is a compound with activity, by way of indication and not limiting, antihypertensive and / or antidiabetic and / or antidepressant, and preferably is gamma-aminobutyric acid.

By "gamma-aminobutyric acid or GABA" is meant a non-protein amino acid of relevant importance as a brain neurotransmitter, which is considered beneficial for good health maintenance and also acts as a hypotensive, antidiabetic and antidepressant compound.

Any variety of brown rice grains is included in the scope of the process of the invention. Examples of rice varieties that are used in the invention are, although not limited to, commercial varieties of Oryza sativa, such as PRONACA SFL09, INIAP 14, INIAP 15, INIAP 17 and experimental cultivation GO39839.

In a particular embodiment, in step a) the time is 24 hours, the temperature is 28 ° C and the or-oryzanol comprises a content greater than 14 mg per 100 g of dry flour of 30 grains of sprouted brown rice and a content of GABA greater than 23 mg per 100 g of dried flour from germinated brown rice grains (see FIG 1 and 2).

The process for obtaining the invention, when using grains of brown rice of the variety SFL 09 and is carried out for 24 hours at 28 ° C, allows an increase of -35

oryzanol of 30% with respect to the same non-germinated rice and 57% with respect to the same rice germinated in the absence of light, and a GABA increase of 2500% with respect to the same non-germinated rice and 92% with respect to the same rice germinated in darkness.

 5

It is the second object of the invention, the product obtained by the process for obtaining the invention, hereinafter product of the invention.

In one aspect of the invention, the product of the invention are germinated brown rice grains, hereinafter germinated brown rice grains of the invention, and / or the 10 products derived therefrom that comprise biologically active compounds.

The germinated brown rice grains of the invention are consumed directly as a washed and clean fresh vegetable to such an extent that they do not require additional washing when cooking and admitting any type of packaging, for sale of fresh produce, canned food (canned, pickled, between others), frozen or any other presentation of edible vegetable foods.

In the present invention, "derived products" refers to the germinated brown rice grains of the invention that undergo some physical or chemical transformation, but also to the extracts comprising the bioactive compounds present in the germinated or modified brown rice grains.

The germinated rice grains of the invention are dehydrated and converted into flours following any conventional procedure of those that are collected in the state of the art, and are used as instant food, flour to batter or prepare bakery foods or pastries, which are additionally suitable for coeliacs.

The sprouted brown rice grains of the invention and the flours obtained therefrom constitute a source of bioactive compounds that are extracted therefrom using appropriate techniques, such as those for example indicated in documents CN102464696 A, CN103082171 A, JP2006111583 A2, JP5070361 B1.

In another aspect of the invention, the product of the invention is an extract that is obtained from the germinated brown rice grains of the invention or from the flours obtained therefrom, which comprises or-oryzanol and / or GABA

It is the third object of the invention, the use of the product of the invention, in the elaboration of healthy food compositions comprising it.

"Healthy food compositions" means solid or liquid food preparations, including food supplements, rich in 5 biologically active compounds, preferably or-oryzanol and / or GABA, which in addition to the proper nutritional value of rice, promote the good state of health and well-being of consumers, for their participation in the prevention and delay in the appearance of chronic degenerative diseases such as hyperlipidemias, cardiovascular diseases, diabetes, obesity, dementia, menopause, depression and cancer. 10

The healthy effect of the bioactive compounds -oryzanol and / or GABA is sufficiently collected in the state of the art section.

Examples of healthy food compositions are, but not limited to, prepared dishes comprising meats, vegetables, vegetables, salads and side dishes; energy and strengthening drinks intended for populations with specific needs in order to achieve their nutritional and functional revaluation; daily products of batter, tempura, pastries, pastries, energy bars, snacks, bread doughs, pastries and related products; and soups, purees and instant drinks. twenty

When the healthy food compositions are food supplements, they are presented in the form of capsules, in lyophilized form, in liquid form, in the form of pills or in the form of gels comprising the bioactive compounds, which are preferably -oryzanol and / or GABA . 25

It is the fourth object of the invention, the use of the product of the invention, in the preparation of cosmetic compositions comprising it.

Finally, it constitutes the fifth object of the invention, healthy food compositions and / or cosmetic compositions comprising the product of the invention.

Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics. For the person skilled in the art, other aspects, advantages and characteristics of the invention will be derived in part from the

description and in part of the practice of the invention. The following figures and examples are provided by way of illustration, and are not intended to be limiting of the present invention.

BRIEF DESCRIPTION OF THE FIGURES 5

Figure 1. Content in -oryzanol (mg / 100g). (A) dry flour from uncooked brown rice grains; (B) dried flour from germinated brown rice grains obtained by the process of obtaining the invention for 24 h at 28 ° C; (C) dried flour from germinated integrated rice grains obtained in the absence of light. 10

Figure 2. GABA content (mg / 100g). (A) dry flour from uncooked brown rice grains; (B) dried flour from germinated brown rice grains obtained by the process of obtaining the invention for 24 h at 28 ° C; (C) dried flour from germinated brown rice grains obtained in the absence of light. fifteen

EMBODIMENTS OF THE INVENTION

Example 1. Obtaining germinated brown rice grains of the invention using the method of obtaining the invention

To obtain the germinated brown rice grains of the invention, a commercial certified brown rice cultivar SFL 09 marketed and distributed by INDIA - PRONACA (Ecuador) was used.

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50 g of brown rice were washed with distilled water and then sanitized with 250 mL of 0.1% sodium hypochlorite for 30 min. They were then drained and washed with distilled water until neutral pH. Subsequently, the rice was germinated in the same amount of distilled water (ratio 1: 5 p / v) and kept for 24 hours at 28 ° C in the presence of light. 30

After that time, the germinated brown rice grains of the invention were drained and lyophilized.

Example 2. Obtaining germinated brown rice grains using a germination procedure in the absence of light.

Identical conditions were reproduced to those indicated in Example 1, but in this case germination was carried out in dark conditions.

Example 3. Analysis of -oryzanol by HPLC in the germinated brown rice grains obtained according to Examples 1 and 2, and in the same non-germinated brown rice 5

The extraction of -oryzanol from samples obtained according to Examples 1 and 2 was carried out with 100% methanol (1:10 ratio, w / v), as a control the unrminated brown rice was used, henceforth raw. For this, 2 g of the corresponding dry rice flour obtained by lyophilization were combined with 20 mL of methanol and vortexed for 2 min. 10 They were subsequently centrifuged at 4500xg, 10 min, at room temperature. The supernatant was removed and the residue was subjected to extraction under the same conditions twice more. The supernatants were bound and concentrated to dryness on a rotary evaporator, at 35 ° C. The obtained residue was dissolved in 1 mL of methanol, filtered through 0.22 µm membranes and its content analyzed by HPLC. The chromatographic system consisted of an Alliance 2695 separation module 15 (Waters, Milford, USA) equipped with a C18 column (150 x 3.9 mm id, 5 µm) (Waters) thermostated at 25 ° C and a 2996 aligned diode detector (Waters) at 325 nm. 20 µL of sample was injected and the separation was carried out at a flow of 1 mL / min by elution of the following gradient formed by mobile phase A acetonitrile HPLC grade, mobile phase B methanol grade HPLC and mobile phase C bi-distilled water with the 20 gradient indicated in Table 1.

Table 1. Elution gradient for the analysis of or-oryzanol by HPLC

 Time (min)

 Phase A (%) Phase B (%) Phase C (%)

 0

 60 35 5

 5

 60 35

 5

 8

 60 40 0

 10

 60 40 0

 twenty

 22 78 0

 35

 22 78 0

 Four. Five

 60 35 5

 fifty

 60 35 5

The identification was made by comparing the maximum absorption wavelength and mass spectrum with a commercial pattern of or-oryzanol (Cymit, Spain). The quantification was performed using a calibration line with the same commercial standard in a concentration range between 5-150 µg / mL (R2> 0.990).

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Example 4.- GABA analysis in the germinated brown rice grains obtained according to Examples 1 and 2, and in the same un germinated brown rice

GABA extraction of the samples obtained according to Examples 1 and 2 was carried out in distilled water (1:20 ratio, w / v), as raw brown rice was used as control. For this, 10 500 mg of the corresponding dry rice flour obtained by lyophilization were combined with 12 mL of water and the mixture was kept under stirring at 4 ° C overnight. They were then centrifuged at 32500xg, 10 min, at 5 ° C. 50 μL of the supernatant was taken and lyophilized, to which 10 μL of internal allyl-glycine standard (1.2 mg / mL) and 20 μL of the methanol: water: triethylamine (1: 1: 0.5) mixture reagent were added ). The mixture was stirred vigorously in vortex and concentrated to dryness. Then 30 µL of the mixture reagent formed by MetOH: phenylisothiocyanate: triethylamine: water (7: 1: 1: 1) was added and allowed to react 20 min at room temperature. Then, its concentration was carried out to dryness and its subsequent dissolution in mobile phase A formed by 0.1 M ammonium acetate, pH 6.5, vigorous mixture in vortex and centrifugation at 32500xg for 5 min. Subsequently, the sample was filtered through 0.22 µm membrane and its content analyzed by HPLC. The chromatographic system consisted of an Alliance 2695 separation module (Waters, Milford, USA) equipped with an Alltima C18 Column column (5 m, 250 x 4.6 mm) preceded by an Alltima C18 Pre-column (5 m, 7.5 x 4.6 mm ) thermostatized at 43 ° C and a 2996 aligned diode detector (Waters) at 242 nm. 20 µL of sample was injected and separation was performed at a flow rate of 0.7 mL / min by elution of the next gradient formed by mobile phase A 0.1 M ammonium acetate, pH 6.5 and mobile phase B ammonium acetate 0, 1 M-acetonitrile-methanol pH 6.5 with the gradient indicated in Table 2.

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Table 2. Elution gradient for GABA analysis by HPLC

 Time (min)

 Phase A (%) Phase B (%)

 0

 100

 0

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 90 100

 40

 60

 40

 42

 0 100

 48

 0 100

 49

 100 0

 53

 100 0

The identification was made by comparing the maximum absorption wavelength and mass spectrum with a commercial GABA pattern (Sigma, Spain). The quantification was performed using an internal and external calibration line with the same commercial standard (allyl-glycine and GABA, respectively) in a concentration range between 15-240 µg / mL (R2> 0.990).

Claims (12)

1. Procedure for obtaining brown rice grains and / or products derived therefrom that comprise biologically active compounds, characterized in that it comprises: a) in the presence of light, germinate brown rice grains for a period of time between 20 and 120 h and at a temperature between 25 and 34 ° C. 2. Method according to claim 1, characterized in that the biologically active compounds are chosen from a compound with antioxidant activity and / or a compound with antihypertensive and / or antidiabetic and / or antidepressant activity. 3. Method according to claim 2, characterized in that the compound with antioxidant activity is or-oryzanol. fifteen 4. Method according to any one of claims 2 and 3, characterized in that the compound with antihypertensive and / or antidiabetic and / or antidepressant activity is gamma-aminobutyric acid.  twenty 5. Method according to any one of claims 1 to 4, characterized in that in step a) the time is 24 hours and the temperature 28 ° C. 6. Method according to claim 5, characterized in that the or-oryzanol comprises a content greater than 14 mg per 100 g of dried flour of germinated brown rice grains and a content of gamma-aminobutyric acid greater than 23 mg per every 100 g of dried flour from germinated brown rice grains. 7. Product obtained by the process as described in any one of claims 1 to 6. 30 8. Product according to claim 7, which are grains of sprouted brown rice. 9. Product according to claim 7, which is a flour obtained from the germinated brown rice grains according to claim 8. 35 10. Use of the product as described in any one of claims 7 to 9, in the preparation of healthy food compositions. 11. Use of the product as described in any one of claims 7 to 9, in the preparation of cosmetic compositions. 5 12. Healthy food compositions and / or cosmetic compositions obtained as described in any one of claims 10 and 11.