Description:Field of Invention
The present invention is directed to a composition obtained from Okra plant for use in enhancing endurance among subjects. The present invention also relates to a method of preparation of said composition and its use for enhancing endurance.
Background
Okra, scientifically known as Abelmoschus esculentus, belonging to the family Malvaceae, is commonly known as Lady’s finger, as well as by several vernacular names, including okra, bhindi, okura, quimgombo, bamia, gombo, and lai long ma, in the different geographical regions of its cultivation. The immature green pods of okra are usually consumed as vegetables, while the extract of the pods also serves as a thickening agent in numerous recipes for soups, as well as sauces, to augment their viscosity. Generally, okra is a high-value crop because it represents a source of nutrients that are important to human health, e.g., vitamins, potassium, calcium, carbohydrates, dietary fiber, and unsaturated fatty acids such as linolenic and oleic acids. Durazzo et al discloses that extracts of young okra pods have also been reported to display moisturizing and diuretic properties, whereas the seeds of this plant have been reported to possess anticancer and fungicidal properties. Durazzo et al evaluated the bioactive components and beneficial properties of Abelmoschus esculentus (Okra) like antidiabetic activity. Dantas et al discloses the extraction of Okra mucilage and its application as thickeners and food stabilizers. Xia Fangbo et al discloses the antioxidant and anti-fatigue constituents from Okra seeds and skins. Yan Li discloses a composition and anti-fatigue activity of water-soluble okra from Okra stem pectins.
The present invention discloses a palatable powder composition prepared from Okra for enhancing endurance and method of preparation of said palatable powder composition.
Detailed description
The present invention discloses a palatable, free flowing powder composition prepared from Okra (Abelmoschus esculentus). The powder composition comprises polysaccharides and carbohydrates. The present invention also discloses the capability of said powder composition for enhancing endurance to the subject when administered an effective amount of the composition and method of preparation of said palatable powder composition.
Okra, scientifically known as Abelmoschus esculentus, is a flowering plant that is valued for its edible green pods. Okra is low in calories and a good source of dietary fiber. It contains various vitamins and minerals, including vitamin C, vitamin K, and folate. One distinctive feature of okra is the mucilage or slime found in the pods. This substance is released when the pods are cut or cooked. The mucilage is known to have thickening properties and is used in some traditional dishes for this reason. The mucilaginous nature of okra mucilage may cause digestive discomfort for some people. It can affect the texture of the food as it passes through the digestive tract and may lead to bloating or gas in some individuals. In the present invention, a palatable, free flowing, light brown colored powder composition is obtained from Okra which is soluble in water. This powder composition is used for enhancing the endurance.
Endurance is a term used to describe the capacity of the body to sustain prolonged physical activity or to resist fatigue over an extended period. It involves the cardiovascular, respiratory, musculoskeletal, and metabolic systems working together to support the demands of prolonged or repetitive exercise. Endurance activities place a significant demand on the adenosine triphosphate (ATP) energy system, and the body's ability to efficiently produce and regenerate ATP which is essential for sustaining performance. ATP is the primary energy currency in cells. The energy stored in ATP is used for various cellular processes, such as muscle contraction, active transport of ions across cell membranes, and synthesis of macromolecules like proteins and nucleic acids. ATP increases cellular energy through the processes of cellular respiration, substrate-level phosphorylation, and the transfer of high-energy phosphate groups. The energy stored in ATP is released when needed to drive various cellular activities. ATPase, short for Adenosine Triphosphatase, is an enzyme that catalyzes the hydrolysis of ATP (adenosine triphosphate) to ADP (adenosine diphosphate) and inorganic phosphate (Pi). This hydrolysis reaction releases energy that can be used by the cell to perform various functions, such as muscular contraction, active transport of ions across cell membranes, and synthesis of macromolecules like proteins and nucleic acids. ATPases play essential roles in cellular metabolism and energy transfer.
The present invention is directed towards a composition prepared from Okra for enhancing endurance. The composition from Okra increases the ATP level in blood, liver and muscles. Composition also increases the ATPases thereby increasing the energy by dephosphorylation. The composition of the invention decreases Creatine Kinase levels in blood, increases the succinate dehydrogensase (SDH) enzyme level, decreases the lactate dehydrogenase (LDH) in blood, decreases less serum urea nitrogen (SUN), decreases lactic acid (LA) and increases the muscle glycogen.
The composition prepared from Okra is a palatable, free flowing powder composition. The composition is administered in an effective dosage form wherein the composition comprises polysaccharides and carbohydrates for enhancing endurance in a subject in need thereof.
As used herein, the term "subject" refers to the target of administration. It can be a vertebrate, such as a mammal, a fish, a bird, a reptile, or an amphibian. Alternatively, the subject of the herein disclosed methods can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term does not denote a particular age or sex. Thus, adult and new-born subjects, whether male or female, are intended to be covered unless otherwise specified.
As used herein, the terms "effective amount" and "effective dose" refer to an amount that is sufficient to achieve the desired therapeutic result or to have an effect on undesired symptoms but is generally insufficient to cause adverse side effects. One skilled in the art should know that the effective amount can vary from one individual to another due to the external factors and age, sex, diseased state, races, body weight, formulation of the extract, availability of other active ingredients in the formulation and so on.
The composition of the present invention comprises 10-50% Polysaccharides and 30-70% total carbohydrates. In one embodiment, the powder composition comprises 36.7% total carbohydrates and 11.5% polysaccharides.
Compositions of the present invention may also comprise other components, for example, one or more excipients.
In another aspect, the present invention provides use of the composition comprising extract of Okra that improves endurance or performance.
The composition of the present invention can be formulated and the formulation can be prepared in a variety of forms. Non-limiting examples of such forms include capsules, tablets, pills, dispersions, suspensions, solutions, powders, teas, syrup concentrates, bars and chews.
The powder composition of the invention may be included as a compound ingredient in other products. Non-limiting examples of other products include food, medicine, sport beverages, meal replacement drinks, powders, ointments, dietary supplement capsules, tablets, or a simple water additive.
In another embodiment, a method of enhancing endurance in a subject, comprises administering to the subject in need thereof an effective amount of the composition.
In an embodiment, the administration of composition disclosed herein to a subject is via an oral route.
In another embodiment, the composition administered to a mammal is at a dose of about 100 to 2000 mg to a mammal per day.
In yet another embodiment, method of preparation of the free-flowing powder composition from okra is disclosed. Dried Okra pods are powdered and extracted with hydroalcoholic solution at 90oC for 90 minutes. Repeat the extraction for three times. All the filtrates are collected and heated to 70oC to form concentrated mixture. Ethyl acetate is added to the concentrated mixture, stirred for 2 minutes at room temperature and then allowed to settle for 1 hour. Separate the ethyl acetate and aqueous layer. Concentrate the aqueous layer at 85oC to form concentrated aqueous solution. Ethanol is added to the concentrated aqueous solution and the mixture is cooled at 4oC for 24 hours. Mixture is clarified and residue and centrifugate is separated. The centrifugate is concentrated at 95oC and dried to form free flowing powder composition of Okra.
In another embodiment, method of preparation of the free-flowing powder composition from okra is disclosed. Dried Okra pods are powdered and extracted with water at 90oC for 90 minutes. Repeat the extraction for three times. All the filtrates are collected and heated to 60oC. Enzyme pectinase is added to the heated mixture and stirred for 4 hours at same temperature. After 4 hours of enzyme reaction, the mixture is clarified and concentrated under vacuum at 95oC. Concentered wash is spray dried to obtain free flowing powder composition of Okra.
Efficacy of the powdered composition is studied by forced swim test in rats. In one embodiment, rats are administered with Okra extract at 100mg/Kg, 1 hour before swimming test. The immobility time is78 sec compared to normal control group which showed an immobility time of 121 seconds. When rats are administered with okra extract treated with enzyme also showed less immobility time of 81 sec compared to normal control group which showed an immobility time of 121 seconds. Less immobility time indicates better endurance and less fatigue. Okra dried powder showed an immobility time of 115 seconds which is comparable to normal control group.
In another embodiment, efficacy of the powdered composition is studied by evaluating the swimming endurance by weight loaded forced swim test (FST)in ratsto assess endurance capacity. Rats administered with okra extract at a dosage of 100mg/Kg/day for 28 days showed high swimming time of 24 secs compared to normal control group which showed a swimming time of 13 secs. Administering Okra extract treated with enzyme at a dosage of 100mg/Kg/day for 28 days to rats also showed high swimming time of 22 seconds. After administering Okra water extract alone to rats showed a swimming time of 16 seconds, and okra dried powder showed a swimming time of 14 seconds which is comparable to normal control group. A higher swimming time indicates better endurance and less fatigue.
Examples
Method of preparation of Okra extract.
1Kg powdered Okra dried pods were extracted with 15L of water and methanol in 80:20 ratio at 90oC for 90 minutes. After the extraction, first filtrate and first residue were separated. First residue was again extracted with 15L of water: methanol in 80:20 ratio at 90oC for 150 minutes to form second filtrate and second residue. Second residue was again extracted with 15L of water: methanol in 80:20 ratio at 90oC for 150 minutes to form third filtrate and third residue. First, second and third filtrates were combined and heated to 70oC to evaporate methanol part to form concentrated aqueous part. 20 Liters of ethyl acetate was added to the concentrated aqueous part, stirred for 2 minutes at room temperature and allowed to settle for 1 hour. After 1 hour, aqueous and ethyl acetate layers were separated. Washing process with ethyl acetate was repeated two more times. The aqueous layer was separated from ethyl acetate layer and the aqueous part was concentrated at 85oC until the TDS reached 20%. 100 liters of ethanol was added slowly to the concentrated aqueous solution and the mixture was cooled at 4oC for 24 hrs. After 24 hours, mixture was clarified and centrifugate and residue were separated. Centrifugate was concentered under vacuum at 95oC until it reached a TDS concentration of 20%. Concentrated wash was spray dried to obtain free flowing powder composition of Okra with 50% yield (sample 1).
Evaluation of swimming endurance of Okra extracts by forced swim test (FST) in rats
Adult male albino rats (Sprague-Dawley) weighing 200- 250g were housed in polypropylene cages. The rats were kept at 22–24°C and 65% relative humidity with alternate light and dark periods of 12-h intervals each. Animals had free access to pellet food and filtered water. The animals were acclimatized for one week before the start of the experiment. Extracts were orally fed to rats at 100 mg/kg, 1 hour before swimming test. Animals were divided into 6 groups with 5 rats in each group. The test included exposure of the animals to a water tank (height, 70 cm; diameter, 40 cm, containing water up to 50 cm of height at 25°C) for 5 minutes duration. Each animal made vigorous attempts to get out of water during the first couple of minutes and thereafter surrendered to experimental conditions and assumed a typical immobile posture (which is defined as when no additional activity was observed other than that required to keep the head above the water) with occasional escape attempts. The total duration of immobility in the test session was recorded. In this test, less immobility time indicates better endurance/less fatigue.
Groups Immobility time (Sec)
Normal control 121
Positive control 78
Okra dried powder 115
Okra water extract 93
Okra water extract+enzyme 81
Okra extract (sample 1) 78

Increased immobility time is often interpreted as a sign of depressive-like behavior in rodents. Conversely, a reduction in immobility time is considered a potential indicator of an antidepressant effect. Endurance is the ability to sustain physical activity over an extended period. Fatigue is the decline in physical and mental performance that occurs after prolonged exertion. In the context of the force swim test, a rat with better endurance would be able to swim and remain active for a longer duration before showing signs of fatigue. In the forced swim test, less immobility time can be associated with better endurance. This is because reduced immobility time implies that the animal is more actively attempting to escape or cope with the stressful situation of being placed in water. Increased activity, such as swimming and struggling, indicates a higher level of endurance or resilience in the face of a challenging environment.
Rats treated with Okra water extract showed an immobility time of 93 seconds compared to normal control group. Group 6 rats treated with sample 1 prepared as per example 1 showed less immobility time of 78 seconds compared to normal control group which showed an immobility time of 121 seconds. Administering Okra extract treated with enzyme at a dosage of 100mg/Kg to rats (group 5)1 hour before swimming test showed less immobility time of 81 sec compared to normal control group which showed an immobility time of 121 seconds. Okra dried powder treated group showed an immobility time of 115 seconds which is comparable to normal control group. Less immobility time indicates better endurance and less fatigue.
Evaluation of swimming endurance of Okra extracts by (weight-loaded) forced swim test (FST) in rats
Adult male albino rats (Sprague-Dawley) weighing 200- 250g were housed in polypropylene cages. The rats were kept at 22–24°C and 65% relative humidity with alternate light and dark periods of 12-h intervals each. Animals had free access to pellet food and filtered water. The animals were acclimatized for one week before the start of the experiment. Rats were divided into 6 groups and each group contain 5 rats. Extracts were orally fed to rats at 100 mg/kg/day for 28 days. After 30 min of last dose, rats were taken out from each group for a weight-loaded swimming test. 30 min after the last intragastric administration, the rats were placed individually in a swimming pool (height, 70 cm; diameter, 40 cm, containing water up to 50 cm of height at 25°C), in which the rats could not support themselves by touching the bottom with their tails. The rats were loaded with a metal block (approximately 5% of body weight) attached to their tails. The swimming time to exhaustion was used as the index of the forced swimming capacity. More swimming time indicates better endurance/less fatigue.
Groups Swimming time (Sec)
Normal control 13
Positive control 25
Okra dried powder 14
Okra water extract 16
Okra water extract +enzyme 22
Okra extract (sample 1) 24

The forced swim test with a weight load is designed to impose a greater physical challenge on the animals, simulating conditions that may induce fatigue more quickly. Rats that exhibit extended swimming times under these challenging conditions are considered to have better endurance and a higher threshold for fatigue. Group 6 animals treated with Okra extract prepared as per example 1 at a dosage of 100mg/Kg/day for 28 days showed high swimming time of 24 seconds compared to normal control group which showed a swimming time of 13 secs. Administering Okra extract treated with enzyme at a dosage of 100mg/Kg/day for 28 days to rats also showed high swimming time of 22 seconds. After administering Okra water extract alone to rats showed a swimming time of 16 seconds. Okra dried powder treated group showed a swimming time of 14 seconds which is comparable to normal control group. A higher swimming time indicates better endurance and less fatigue. , Claims:.