A functional beverage containing bitter apricot kernel extract and a method for its preparation

Abstract

The invention relates to a new functional beverage with a high phenolic compound content for use in the nutrition and food industry and a method for its preparation. The invention particularly relates to a functional beverage comprising bitter apricot kernel extract, which has a high content of bioactive compounds, having antioxidant, antigenotoxic, antihyperglycemic and antihyperlipidemic effects, and a method for its preparation. The beverage in question is a type of coffee beverage.

Description

[0001] A FUNCTIONAL BEVERAGE CONTAINING BITTER APRICOT KERNEL EXTRACT AND A METHOD FOR ITS PREPARATION

[0002] Field of the Invention

[0003] The invention relates to a new functional beverage with a high phenolic compound content for use in the nutrition and food industry and a method for its preparation.

[0004] The invention in particular relates to a functional beverage comprising bitter apricot kernel extract, with a high content of bioactive compounds, having antioxidant, antigenotoxic, antihyperglycemic and antihyperlipidemic effects, and a method for its preparation. The beverage in question is a type of coffee beverage.

[0005] State of the Art

[0006] Since they are rich in active metabolites, plant extracts have been widely used for centuries in the treatment of diseases among the people. Phytochemicals are non- essential primary or secondary metabolites found in the flowers, leaves, stems, and bodies of plants, which are produced by plants, and playing a role in their defense mechanisms, they provide plants with color, taste, smell, and aroma and possess strong antioxidant activity that is protective or disease preventing. Although it is known that plants produce these chemicals to protect themselves, recent research shows that these chemicals have protective effects against diseases. Phytochemicals and their derivatives have been shown to be beneficial for human health due to their antioxidant, antiinflammatory and anti-proliferative properties. Although secondary metabolites do not have a direct role in the vital functions of life in living creatures, they have an important role in the protection and maintenance of health and are used as raw materials in many industries today. In recent years, it is known that polyphenols, flavonoids and phenolic acids in the contents of plants have antigenotoxic, antidiabetic, antihyperlipidemic and anti-inflammatory effects by cleaning reactive oxygen species (ROS) and increasing host antioxidant defense systems. In addition, numerous studies showing that phytochemicals protect against DNA damage have revealed the presence of strong phyto-antimutagens in the plant kingdom.

[0007] The fruits of the apricot tree (Prunus armeniaca L.), a member of the Rosaceae family, are rich in dietary fibers, proteins, sugars, fatty acids, micronutrients, volatile compounds, carotenoids, phenolics, and lignans. Moreover, its various parts are widely used in the treatment of various diseases including respiratory diseases, digestive disorders and gynecological diseases due to its various biological properties such as antipyretic, wound healing, anti-inflammatory, antidiabetic, antihelminthic, he pato protective and anticancer activity. Apricot kernels contain a significant amount of fats, carbohydrates, proteins, polyphenols, flavonoids, fibers and micronutrients such as cyanogenic glycoside amygdalin, vitamins (A, B group, C, E and K), amino acids and fatty acids, and it has been shown to have a great effect on human health as it can be beneficial for intestinal health and cholesterol absorption, as well as having anti-obesity and anti-diabetes properties. In addition, various parts of the apricot plant are used in clinical trials of cancer treatment in the field of complementary medicine worldwide, and apricot based products have shown promising therapeutic potential in this field. It has been stated that the proteins found in apricot kernels increase the antioxidant activity of foods and that apricot kernels comprise various types of antioxidant components such as lutein, beta carotene and zeaxanthin, which also help fight free radicals.

[0008] Apricot kernels are classified as sweet apricot, semibitter apricot and bitter apricot according to their taste. Apricot kernels comprise cyanogenic glycoside amygdalin (up to 4.9%) as the main component, and these cyanogenic glycosides are especially present in high amounts in bitter apricot kernels.

[0009] Various studies in the literature show that apricot kernels have biological properties such as protective activity on the heart, antioxidative / radical scavenging, neuroprotective, antihyperlipidemic, he pato protective, antimicrobial, antiparasitic, antiviral, antiinflammatory, antidiabetic, analgesic, immunomodulatory and anticancer activity.

[0010] Coffee is the most important commercial food product worldwide, and most of the coffee drinks consumed worldwide are produced as Arabica and Robusta. The coffee plant, botanically classified within the Rubiaceae family, encompasses approximately 80 species. 2 species, Arabica and Robusta, are cultivated worldwide. 75% of the global coffee production is covered by Arabica beans and 24% by Robusta beans. Coffee fruits are harvested after their color changes from green to red, and the beans are separated from the pulp by pulp separation machines so that the beans, which are thinly coated on the outside, can be taken out. After the other layers other than the coffee bean membrane are separated by processing with wet or dry processing techniques, the coffee beans are dried. In the roasting stage, where the coffee gains color and aroma, green coffee beans are roasted at 210- 24012 and the water is evaporat ed. Studies in the literature have shown that coffee exhibits antioxidant, anti-inflammatory and anti-mutagenic properties depending on the bioactive components such as caffeine, chlorogenic acid and diterpenes it contains. Regarding the effects of coffee on glucose homeostasis; caffeine has been shown to reduce insulin sensitivity due to adrenaline release. Adenosine A1 receptor antagonism resulting from acute coffee intake (within 1- 3 hours) may reduce skeletal muscle glucose uptake. In addition, coffee polyphenols have important antioxidant and anti-inflammatory effects that may affect the glucose metabolism and insulin sensitivity.

[0011] The improper use of these plant extracts, which have benefits in different areas, in daily life can cause various toxic effects on the human body. Amygdalin (D-mandelonitrile-b- gentiobioside) is one of the main bioactive substances found not only in apricot kernels, but also in products such as bitter almonds, flaxseeds, etc. Amygdalin is a natural compound used in traditional medicine as an anti-cancer, antipyretic and cough suppressant. Today, it is known to be applied for the prevention and treatment of migraine, hypertonia, chronic inflammation, asthma, bronchitis, emphysema, leprosy, diabetes, colorectal cancer and vitiligo. In a study evaluating the antioxidant effects of amygdalin on tunicamycin induced endoplasmic reticulum stress in mouse livers, it was found that pretreatment with amygdalin reduced tunicamycin induced endoplasmic reticulum stress and had a significant antioxidant activity in liver tissue. In a study, the in vitro genotoxic and anti genotoxic effects of amygdalin on human peripheric blood lymphocytes were evaluated by comet assay and found to have an anti-genotoxic effect against H2O2. Today, this compound is called vitamin B17 and is found in about 3% of apricot kernels. Considering it is a bioactive substance with highly beneficial content, there might be a desire to consume it in large amounts, but controlled consumption enhances its effectiveness.

[0012] Nutrition and stress management are very important in terms of improving the quality of life in both healthy individuals and chronic and metabolic diseases (diabetes, cancer, thyroid, etc.). In line with the increasing interest in fibrous, high nutritional value, and antioxidant comprising foods, the need for functional foods in the society is increasing. There is an uneasy approach to the consumption of natural food sources containing B17, which is very important to consume in a controlled manner. For these reasons, in the state of the art, there is a need for a functional food in which B17 is taken in an effective and appropriate dose that can be added to the daily diet in healthy individuals and different disease models. When evaluated in terms of its protective properties in the technique, there is no product that allows two strong contents such as coffee and bitter apricot kernel extract to be taken into the body at the same time.

[0013] As a result due to the said disadvantages and the insufficiency of the current solutions regarding the subject matter, a development is required to be made in the relevant technical field.

[0014] Objective of the Invention

[0015] The present invention relates to a new functional beverage with a high content of bioactive compounds and a method for preparing the same, using bitter apricot kernel extract, which meets the above-mentioned requirements, eliminates all disadvantages and brings some additional advantages.

[0016] The main object of the invention is to provide a functional beverage rich in bioactive content that will be effective in the intake of vitamin B17 (Amygdalin) in an effective and appropriate dose of bitter apricot kernel extract and to produce this beverage.

[0017] The object of the invention is to provide a product with antioxidant, antigenotoxic, antihyperglycemic and antihyperlipidemic effects that will provide numerous benefits in different disease models with bitter apricot kernel extract and coffee content.

[0018] Another object of the invention is to ensure that the active ingredients in the kernels are taken into the body in a controlled manner with this functional and practical beverage obtained by mixing apricot kernel extract and coffee. Thus, it is ensured that bitter apricot kernels, which will be preferred in a more limited manner, are added to the social life routine against the possibility of toxic effects when used alone.

[0019] Another object of the invention is to provide a product that combines the biological properties of apricot kernels such as protective activity on the heart, antioxidative / radical scavenging, neuroprotective, antihyperlipidemic, hepatoprotective, antimicrobial, antiparasitic, antiviral, anti-inflammatory, antidiabetic, analgesic, immunomodulatory and anticancer activity with the antioxidant, anti-inflammatory and anti-mutagenic properties of coffee.

[0020] Another object of the invention is to provide a product that will provide protection against diseases with the effects of natural bioactive components (Amygdalin, caffeine, chlorogenic acid and diterpenes) in its content and to ensure that these products are easily accessible to individuals.

[0021] In order to fulfill the aforementioned objects,, the invention is a is a functional beverage with a high bioactive content, comprising Bitter Apricot Kernel Extract powder and ground coffee.

[0022] In order to fulfill the aforementioned objects, the invention is a method of preparing a functional beverage with a high bioactive content comprising the following process steps: i. Weighing the bitter apricot kernel ii. Extraction of Bitter Apricot Kernels with Solvents and then Removal of the Solvent iii. Lyophilization drying and powdering of the Bitter Apricot Kernel Extract, iv. Mixing Bitter Apricot Kernel Extract Powder with ground coffee.

[0023] The structural and characteristic features, as well as all advantages of the invention, will be clearly understood through the detailed description provided below and references made to the accompanying figures, therefore, evaluation should be based on both the figures and the detailed description.

[0024] Figures for a Better Understanding of the Invention

[0025] Figure 1, SRB test results of cytotoxic effects of the combination of Doxorubicin (DOX), Methotrexate (MET) and 5-Fluorouracil (5FU) and BAE (Bitter apricot kernel extract) on MDA-MB-231 and MCF-7 breast cancer cell lines of BAE.

[0026] Detailed Description of the Invention

[0027] In this detailed description, the preferred embodiments of the invention are described solely for the purpose of a better understanding of the subject matter and without any limiting effect.

[0028] The invention provides a new approach to producing a beverage, preferably a beverage which is rich in bioactive content and whose uptake of this content is increased.

[0029] A functional beverage with a high bioactive content, which is the subject of the invention, comprises Bitter Apricot Kernel Extract powder and ground coffee. Said functional beverage preferably comprises 20% bitter Apricot Kernel Extract powder by weight and 80% ground coffee by weight.

[0030] In the Bitter Apricot Kernel Extract product of the invention, effective and non-toxic dose of 20% is preferred.

[0031] The product of the invention is in the form of a homogeneous powder. It is preferably in ready to consume form such as sachet form and can be consumed by preparing in boiled water.

[0032] A preferred embodiment of the product of the invention includes Bitter Apricot Kernel Extract powder and ground Arabica coffee.

[0033] In order to achieve said objects, the invention is a method of preparing a functional beverage with a high bioactive content comprising the following process steps: i. Weighing of the bitter apricot kernel ii. Extraction of Bitter Apricot Kernels with Solvents and Subsequent Removal of the Solvent iii. Lyophilization drying and powdering of the Bitter Apricot Kernel Extract Powder Form, iv. Mixing Bitter Apricot Kernel powder extract with ground coffee.

[0034] In order to obtain a preferred example of the invention, the following processes are followed:

[0035] (i) Weighing of the bitter apricot kernels in the desired ratio

[0036] (ii) The weighed Bitter Apricot Kernels are extracted by sequential extraction using ethanol and water and then the solvent is removed by evaporator..

[0037] (iii) The bitter Apricot Kernel Extracts obtained here are dried by lyophylisation for preferably 48 hours to give them a powder form. Here, the extracts are kept at -80 C in case they are kept before lyophylisation (freeze-drying) process.

[0038] (iv) Bitter Apricot Kernel powder extract is mixing with ground coffee in a ratio of 1 :4.

[0039] In our method of the invention, the following points should be considered when doing the extraction. 1- The temperature is adjusted after placing the samples with the appropriate solvent.

[0040] 2- The solvent should be added in sufficient quantity to perform solvent siphoning. Care should be taken to ensure that there is no leakage.

[0041] 3- The cooling system should be adequate in quantity.

[0042] 4- After extraction is completed, the system should be allowed to cool down.

[0043] 1 gram of BAE should comprise 3% Amygdalin, and the BAE extract combined with coffee should be prepared in the appropriate dosage determined.

[0044] The coffee comprising bitter apricot kernel extract of the invention, is the first as a functional beverage, and considering its rich contents, especially phenolic compounds, in both components, antioxidant, antigenotoxic, antihyperglycemic and antihyperlipidemic effects are observed. The following analyzes were applied to determine the effects on the product of the invention; Cytotoxicity tests (SRB) related to the effects of Coffee + BAE (Bitter Apricot Kernel Extract) on in vitro breast cancer cell lines, as well as analyses of glucose, insulin, GSH-Px , SOD, LDH, LDL, fodder, fluid, and body weight. Antioxidant tests (DPPH and CUPRAC) of coffee and BAE were performed. Additionally, a diabetes model was established in vivo using experimental animals.

[0045] Studies on Coffee+BAE beverage, BAE (Bitter apricot kernel extract), Coffee, which is the product of the invention, are given below;

[0046] 1. The antioxidant tests DPPH ((2,2-diphenyl-1-picrylhydrazyl)and CuPRAC ("CupricReducing Antioxidant Capacity", "copper(ll) reducing antioxidant capacity") were performed. The ratios used are given below. IC50 values of BAE and coffee+BAE were used in antioxidant tests. Trolox IC50 5.86 ug / ml

[0047] As a result of the tests, it was determined that the coffee+ BAE mixture of the invention had a very high antioxidant effect.

[0048] 2. To determine its effectiveness in cancer, one of the selected chronic disease models, cytotoxicity tests (SRB) were conducted on in vitro breast cancer cell lines to evaluate the effects of BAE (Bitter Apricot Kernel Extract). According to the SRB test performed with two different cancer cell lines (MCF-7, MDA-MB-231) selected here, combination treatments of BAE were performed with Doxorubicin (DOX), Methotrexate (MET) and 5- Fluorouracil (5FU) used in traditional treatment. Figure 1 shows the cytotoxic effects of BAE (Bitter Apricot Kernel Extract) alone and in combination with Doxorubicin (DOX), Methotrexate (MET), and 5-Fluorouracil (5FU) on MDA-MB-231 and MCF-7 breast cancer cell lines, as determined by SRB assay results.

[0049] As a result of the test, it was determined that the coffee+ BAE mixture of the invention had synergistic and / or antagonistic effects at different doses with all three cancer drugs. Table-1 shows the Cl values of the cytotoxic effects of BAE on MDA-MB-231 and MCF- 7 breast cancer cell lines according to the SRB test.

[0050] Table 1 shows the Cl values of BAE on MDA-MB-231 and MCF-7 breast cancer cell lines as determined by the SRB assay for cytotoxic effects.

[0051] In the trials evaluating the effect of BAE on in vitro breast cancer cell lines; a doseresponse relationship was assessed using arithmetic fold increase over a wide dose range of 0 to 1000 pg / ml.

[0052] A diabetes model was established in vivo using experimental animals. The possible benefits of the BAE in diabetes have been evaluated. Relevant analyses were performed and their positive effects on diabetes were determined according to the results of statistical evaluation.

[0053]

Claims

CLAIMS1. A functional beverage with high bioactive content, characterized by comprising; Bitter Apricot Kernel Extract powder and ground coffee.

2. The beverage according to claim 1 , characterized in that; the said coffee being Arabica coffee.

3. The beverage according to claim 1 , characterized by comprising; 20% bitter Apricot Kernel Extract powder by weight and 80% ground coffee by weight.

4. A method for producing a functional beverage with high bioactive content, characterized by comprising; the following process steps: i. Weighing of the bitter apricot kernels ii. Extraction of Bitter Apricot Kernels with Solvents and subsequent removal of the solvent iii. Lyophilization drying and powdering of Bitter Apricot Kernel Extract, iv. Mixing the Bitter Apricot Kernel Extract powder with ground coffee.

5. The method according to claim 4, characterized in that; in process step ii) Bitter Apricot Kernels are extracted by sequential extraction with ethanol and water as solvents6. The method according to claim 4, characterized in that; in process step ii) after the extraction process in the procedure, the solvent is removed by evaporator.

7. The method according to claim 4, characterized in that; in the process step iii) the Bitter Apricot Kernel Extract is dried by lyophilization for 48 hours.

8. The method according to claim 4, characterized in that; in process step iv) the Bitter Apricot Kernel powder extract is mixed with ground coffee in a ratio of 1 :4 by weight.

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