Compositions and methods for obtaining cellular energy using mint concentrates

Abstract

Compositions comprising a peppermint concentrate are provided. The compositions may be administered to an individual in need thereof for (i) improving a physiological state associated with metabolic fatigue in one or more cells of the individual, and / or (ii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells of the individual, and / or (iii) enhancing mitochondrial function in the individual, and / or (iv) treating or preventing a calcium deficiency / depletion disorder in the individual. Additionally or alternatively, the compositions may be administered to treat or prevent a mitochondrial-related disease or a condition associated with a change in mitochondrial function in an individual in need or at risk.

Description

Technical Field

[0001] This invention relates generally to compositions and methods for managing energy at the cellular level using peppermint concentrate. These compositions and methods can enhance mitochondrial function and increase bioenergetics by activating mitochondrial calcium oneway transporters, thereby promoting cell activation, and in some embodiments, promoting cell activation in middle-aged and elderly individuals, the elderly, or ICU patients. Background Technology

[0002] Population aging has always been a noteworthy demographic event. Due to increased life expectancy, the elderly population is growing faster than the overall population, and due to declining birth rates, the proportion of elderly people relative to other population groups has increased significantly. For example, in 1950, one in twelve individuals was at least 60 years old, and by the end of 2000, one in ten people was 60 years of age or older. By the end of 2050, the number of people aged 60 or older worldwide is projected to be one in five.

[0003] Individuals who are already aging or in the process of aging often experience some degree of physical decline and / or cognitive impairment, including age-related cognitive decline, and age-related changes in brain morphology and cerebrovascular function are commonly observed. Cognitive decline is consistently reported as aging across a range of cognitive domains, including processing speed, attention, episodic memory, spatial ability, and executive function. Brain imaging studies have shown that these normal age-related cognitive declines are associated with a reduction in the volume of gray and white matter in the brain, with the frontal striatum system being most affected by aging. This reduction in cortical volume can be attributed to numerous harmful cellular processes involved in normal aging, such as the accumulation of free radical damage over time leading to oxidative damage, chronic low-grade inflammation, homocysteine ​​buildup (which, when elevated, is a risk factor for cognitive impairment and dementia), and reduced mitochondrial efficiency. In addition to direct cellular damage, the brain is also indirectly impaired by damage to microvascular structures. Clearly, the pathology of aging and dementia involves the complexity of these interrelated factors. For example, mitochondrial dysfunction leads to increased oxidative stress, which can trigger inflammation and vascular damage.

[0004] Mitochondria are the primary source of aerobic energy production in mammalian cells and maintain a large Ca2+ gradient on their inner membrane, providing a signaling potential for the molecule. Furthermore, mitochondrial Ca2+ plays a role in regulating ATP production within mitochondria and potentially contributes to the coordination of cellular metabolic homeostasis (Glancy, B. et al. (2012). “Role of mitochondrial Ca2+ in the regulation of cellular energetics”, Biochemistry, Vol. 14, No. 51: 2959-2973). Alterations in mitochondrial Ca2+ homeostasis have been associated with a variety of pathological conditions and are key to the etiology of several human diseases (Arduino et al., Journal Physiol. July 2018; 596(14):2717-2733).

[0005] Nutrition, education, physical exercise, and cognitive training have recently been demonstrated as potential interventions to prevent age-related physical and cognitive decline. A wealth of clinical, epidemiological, and individual evidence supports individual nutritional factors that reduce the risk of dementia and age-related neurodegeneration. However, results from formal trial testing of nutritional interventions have been mixed (Schmitt et al., Nutrition Reviews 68: S2–S5 (2010)). Furthermore, efforts have been made to utilize mitochondrial Ca2+ transport mechanisms for therapeutic interventions, but pharmacological compounds that guide and selectively modulate mitochondrial Ca2+ homeostasis are currently lacking.

[0006] Therefore, there is a need for compounds that can directly and selectively regulate mitochondrial Ca2+ homeostasis. It is also desirable that such compounds be natural. Furthermore, it is desirable that such compounds be suitable for vegan diets.

[0007] Any references to prior art documents in this specification should not be construed as an admission that such prior art is well-known or part of common knowledge in the field. Summary of the Invention

[0008] The object of this invention is to improve the prior art, and more specifically, to provide compositions, unit dosage forms, and methods that overcome the problems of the prior art and solve the above-mentioned needs, or at least to provide useful alternatives.

[0009] The inventors were surprised to find that the objective of the invention could be achieved through the subject matter of the independent claims. The dependent claims further expand the conception of the invention.

[0010] In particular, based on experimental data to be published later in this article, it is believed that peppermint concentrates as disclosed herein enhance the efficiency of mitochondrial energy production.

[0011] Therefore, embodiments of the present invention provide compositions comprising an effective amount of peppermint concentrate for (i) improving physiological states associated with metabolic fatigue in one or more cells of an individual, and / or (ii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells of an individual, and / or (iii) enhancing mitochondrial function of an individual, and / or (iv) treating or preventing calcium deficiency / depletion disorders in an individual.

[0012] The peppermint concentrate can be obtained by a method including the following steps or by the method thereof:

[0013] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0014] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0015] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0016] d) Optionally, dried peppermint concentrate.

[0017] In one particular implementation, at least a portion of one or more cells is part of at least one body part selected from the group consisting of the liver, kidney, brain, and skeletal muscle.

[0018] In another specific embodiment, physiological states associated with metabolic fatigue include muscle fatigue, muscle weakness, lack of energy, weakness, or lack of energy, particularly lack of physical energy. In another specific embodiment, an effective amount of peppermint concentrate is administered orally daily for at least one week.

[0019] In an additional specific embodiment, the composition further comprises at least one compound selected from the group consisting of: antioxidants, anti-inflammatory compounds, glycosaminoglycans, prebiotics, fiber, probiotics, fatty acids, enzymes, minerals, trace elements and / or vitamins.

[0020] In another specific embodiment, the composition is selected from the group consisting of: food compositions, dietary supplements, nutritional compositions, complete nutritional compositions, pharmaceuticals, oral nutritional supplements, medical foods, nutritional products, beverages, powdered nutritional products reconstituted in water or milk before consumption, food additives, foods for special medical purposes (FSMP), pharmaceuticals, pet foods, and combinations thereof.

[0021] In other specific embodiments, the composition is in the form of a solid powder, powder rod, capsule, or solution.

[0022] In another specific embodiment, an effective amount of peppermint concentrate is applied to a food product or beverage that also contains components selected from the group consisting of: proteins, carbohydrates, fats, and mixtures thereof.

[0023] Another embodiment of the invention provides a composition comprising peppermint concentrate for use in an effective amount to treat mitochondrial-related diseases or conditions associated with altered mitochondrial function in individuals in need or at risk, reducing their morbidity and / or severity, the method comprising orally administering an effective amount of peppermint concentrate to the individual in need or at risk.

[0024] The peppermint concentrate can be obtained by a method including the following steps or by the method thereof:

[0025] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0026] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0027] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0028] d) Optionally, dried peppermint concentrate.

[0029] In one particular implementation, mitochondrial-related diseases or conditions are selected from the group consisting of: stress, physiological aging, obesity, decreased metabolic rate, metabolic syndrome, diabetes, diabetic complications, hyperlipidemia, neurodegenerative diseases, cognitive impairment, stress-induced or stress-related cognitive dysfunction, mood disorders, anxiety disorders, age-related neuronal death or dysfunction, musculoskeletal disorders, frailty, pre-frailty, chronic kidney disease, kidney failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathy, and malnutrition, as well as combinations thereof.

[0030] Another embodiment of the present invention provides a composition comprising peppermint concentrate for use in effective amounts to delay the onset of metabolic decline in healthy middle-aged and older adults, maintain muscle mass and / or muscle function in healthy middle-aged and older adults, maintain immune function in healthy middle-aged and older adults, and / or maintain cognitive function in healthy middle-aged and older adults.

[0031] The peppermint concentrate can be obtained by a method including the following steps or by the method thereof:

[0032] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0033] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0034] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0035] d) Optionally, dried peppermint concentrate.

[0036] Another embodiment of the invention provides a composition comprising peppermint concentrate for use in an effective amount to enhance at least one of an individual's psychological or muscular performance.

[0037] The peppermint concentrate can be obtained by a method including the following steps or by the method thereof:

[0038] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0039] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0040] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0041] d) Optionally, dried peppermint concentrate.

[0042] Another embodiment of the invention provides a composition comprising peppermint concentrate for use in an effective amount to improve or maintain an individual's cognitive function.

[0043] The peppermint concentrate can be obtained by a method including the following steps or by the method thereof:

[0044] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0045] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0046] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0047] d) Optionally, dried peppermint concentrate.

[0048] In one particular implementation, cognitive functions are selected from the group consisting of: perception, memory, attention, speech comprehension, speech generation, reading comprehension, image creation, learning, reasoning, and combinations thereof.

[0049] In one specific embodiment of all the foregoing embodiments, an effective amount of peppermint concentrate may be administered in a composition that also contains calcium. In one specific embodiment of all the foregoing embodiments, the individual is a middle-aged or elderly person, an elderly person, or an ICU patient.

[0050] Another embodiment of the invention provides a unit dosage form comprising peppermint concentrate for use in an effective amount for at least one of the following: (i) treating mitochondrial-related diseases or conditions associated with altered mitochondrial function, reducing their incidence or severity, and / or (ii) improving physiological states associated with metabolic fatigue in one or more cells, and / or (iii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells, and / or (iv) treating or preventing calcium deficiency / depletion disorders, and / or (v) increasing metabolic rate, and / or (vi) improving or maintaining cognitive function, and / or (vii) increasing or maintaining mitochondrial function.

[0051] The peppermint concentrate can be obtained by a method including the following steps or by the method thereof:

[0052] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0053] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0054] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0055] d) Optionally, dried peppermint concentrate.

[0056] In one particular implementation, physiological states associated with metabolic fatigue include muscle fatigue, muscle weakness, lack of energy, weakness or lack of energy, particularly lack of physical energy.

[0057] Those skilled in the art will gain a clearer understanding of these and other aspects, features, and advantages of the present invention after reading the detailed description of the embodiments of the present invention in conjunction with the accompanying drawings. Attached Figure Description

[0058] Figure 1 The iron content (mg / kg) based on dry weight (DW) is shown in peppermint material A and the corresponding peppermint concentrate A obtained from said peppermint material A according to the method of Example 1. The values ​​represent the average among three independent samples, and the error bars represent the standard deviation. Figure 1 The term "peppermint material" should be understood as the whole of the leaves and stems of dried peppermint material A that has been ground into powder before being further processed in the concentration method of Example 1.

[0059] Figure 2 The molar ratio (M / M) between the iron and oxalic acid molar concentrations in peppermint material A and the corresponding peppermint concentrate A obtained from said peppermint material A according to the method of Example 1 is shown. The value represents the average between two repeated measurements. Figure 2The term "peppermint material" should be understood as the whole of the leaves and stems of dried peppermint material A that has been ground into powder before being further processed in the concentration method of Example 1.

[0060] Figure 3 The molar ratio (M / M) between the iron and phytic acid molar concentrations in peppermint materials A and B, and the corresponding peppermint concentrates A and B (= peppermint A concentrate and peppermint B concentrate) obtained from said peppermint materials A and B respectively according to the method of Example 1, is shown. The values ​​represent the average between two independent samples, and the error bars represent the standard deviation. Figure 3 The terms “peppermint material A” and “peppermint material B” should be understood as the whole of the respective leaves and stems of dried peppermint materials A and B, which are ground into powder before being further processed in the concentration method of Example 1.

[0061] Figure 4 Iron bioavailability is shown for a peppermint concentrate prepared from peppermint material B using water (according to the method of Example 1) and a peppermint concentrate prepared from peppermint material B using water in the presence of citric acid, hydrochloric acid, malic acid, or ascorbic acid (according to the method of Example 2). Values ​​represent the average between two independent samples, and error bars represent the standard deviation.

[0062] Figure 5 The absolute amounts of bioavailable iron contained in peppermint material B and the corresponding peppermint concentrate are shown. The peppermint concentrate was prepared from peppermint material B with water according to the concentration method of Example 1, or from peppermint material B with water in the presence of citric acid, hydrochloric acid, malic acid, and ascorbic acid, respectively, according to the concentration method of Example 2. The absolute amount of bioavailable iron was calculated by multiplying the iron content in the sample by the bioavailability value. The value represents the average between two independent samples, and the error bars represent the standard deviation. Figure 5 The term "peppermint material" should be understood as the whole of the leaves and stems of dried peppermint material B that have been ground into powder before being further processed in the concentration method of Example 1 or Example 2.

[0063] Figure 6This figure illustrates the effect of different concentrations of peppermint concentrate B (= peppermint B concentrate) on mitochondrial Ca2+ uptake (= mitochondrial Ca2+ elevation). The graph shows the effects of different dilutions of peppermint concentrate B (light gray 15.6 μg / ml, gray 31.25 μg / ml, and dark gray 62.5 μg / ml) or the control (ctrl). Data are plotted as the percentage change in the area under the curve (AUC) of the integrated mitochondrial calcium elevation induced by 5 mM caffeine stimulation, normalized relative to a 100% control (aqueous buffer). Results are presented as mean + / - SEM from n=8 biological replicates. A statistically significant difference between conditions is indicated by P < 0.05 (one-way ANOVA).

[0064] Figure 7 This figure illustrates the effect of in vitro digests of peppermint concentrate B (= peppermint B concentrate) at different concentrations on mitochondrial Ca2+ uptake (= mitochondrial Ca2+ elevation) in C2C12-derived myotubes. The figure shows the effects of different concentrations of in vitro digests of peppermint concentrate B (light gray for 15.6 μg / ml, gray for 31.25 μg / ml, and dark gray for 62.5 μg / ml). Data are plotted as the percentage change in the area under the curve of integrated mitochondrial calcium elevation induced by 5 mM caffeine stimulation, normalized relative to a 100% control (aqueous buffer). Results are presented as mean + / - SEM from n=8 biological replicates. A statistically significant difference between conditions is indicated by P < 0.05 (one-way ANOVA). Detailed Implementation

[0065] As used herein, the terms “comprising,” “including,” etc., should be interpreted as inclusive, the opposite of exclusive or exhaustive, meaning “including but not limited to.” Similarly, the terms “comprising / including” and “or” should be considered inclusive unless the context explicitly prohibits this interpretation. However, the compositions disclosed herein may not contain any elements not specifically disclosed herein. Therefore, the disclosure of embodiments using the term “comprising / including” includes both embodiments “consisting substantially of the specified components” and embodiments “consisting of the specified components.”

[0066] As used herein, “composition consisting essentially of peppermint concentrate” and “composition consisting essentially of calcium peppermint concentrate” do not include any additional compounds affecting mitochondrial calcium input other than the peppermint concentrate disclosed herein and optional calcium. In one particular non-limiting embodiment, the composition consists of an excipient, the peppermint concentrate disclosed herein, and optional calcium.

[0067] All numerical ranges should be understood to include every integer within the range, whether whole or fractional. Furthermore, these numerical ranges should be understood to support claims involving any number or subset of numbers within the range. For example, disclosures of 1 to 10 should be understood to support ranges of 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, etc.

[0068] All percentages expressed herein are by weight of the total weight of the composition unless otherwise stated. As used herein, “about,” “approximately,” and “substantially” should be understood to mean values ​​within a certain numerical range, such as -10% to +10% of the mentioned values, preferably -5% to +5%, more preferably -1% to +1%, and most preferably -0.1% to +0.1%. All numerical ranges herein should be understood to include all integers or fractions within that range.

[0069] As used herein, the singular forms “a,” “an,” and “the” include multiple referents unless the context explicitly indicates otherwise. Thus, for example, references to “a bioactive compound” or “the bioactive compound” include one bioactive compound but also two or more bioactive compounds.

[0070] Unless otherwise stated, all percentages in this specification refer to weight percentages where applicable.

[0071] Unless otherwise defined, all technical terms have and should be given the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0072] The term “and / or” used in the context of “X and / or Y” should be interpreted as “X” or “Y” or “X and Y”. Similarly, “at least one of X or Y” should be interpreted as “X” or “Y” or “both X and Y”. For example, “at least one of mental or muscular expression” means “mental expression” or “muscular expression” or “both mental and muscular expression”.

[0073] As used herein, the terms “example” and “such as” (especially when followed by a list of terms) are exemplary and illustrative only and should not be considered exclusive or comprehensive. However, disclosure of embodiments using the terms “example” and “such as” includes disclosure of embodiments in which “the terms are exclusive and / or comprehensive”.

[0074] As used herein, “associated with” and “related to” mean that they occur simultaneously, preferably caused by the same underlying disease, and most preferably caused by one of the identified diseases being caused by the other identified disease.

[0075] As used herein, the terms “food,” “food product,” and “food composition” mean a product or composition intended for ingestion by an individual (such as a human) and to provide that individual with at least one nutrient. Compositions of this disclosure (including the various embodiments described herein) may comprise, consist of, or consist substantially of the elements disclosed herein, and any additional or optional ingredients, components, or elements described herein or intended for use in a diet.

[0076] As used herein, the terms “beverage” and “drink” refer to “food,” “food product,” or “food composition” that is typically consumed by drinking. In particular, the terms “beverage” and “drink” are used interchangeably.

[0077] As used herein, the term "treatment" means applying the compositions disclosed herein to a subject suffering from a condition to alleviate, reduce, or improve at least one symptom associated with the condition and / or slow, reduce, or halt the progression of the condition. The term "treatment" includes both preventative or preventive treatment (preventing and / or delaying the development or progression of a target pathological condition or disorder), as well as curative, therapeutic, or disease-modifying treatment, including therapeutic measures to cure, delay, or alleviate symptoms of a diagnosed pathological condition or disorder and / or halt its progression; and treatment of patients at risk of or suspected of having the disease, and treatment of patients who are ill or have been diagnosed with a disease or medical condition. The term "treatment" does not necessarily mean that a subject is treated until fully recovered. The term "treatment" also refers to health maintenance and / or promotion in individuals who do not have the disease but may be prone to developing unhealthy conditions. The term "treatment" is also intended to include intensifying or otherwise enhancing one or more major preventative or therapeutic measures. As a non-limiting example, treatment may be administered by a patient, caregiver, physician, nurse, or other healthcare professional.

[0078] Both human and veterinary treatments are within the scope of this disclosure. Preferably, the peppermint concentrate disclosed herein is administered in portions or units providing an effective amount for treatment or prevention.

[0079] As used herein, the term "prevention" means applying the compositions disclosed herein to a subject who does not exhibit any symptoms of the condition in order to reduce or prevent the development of at least one symptom associated with the condition. Furthermore, "prevention" includes reducing the risk, morbidity, and / or severity of the condition or disorder.

[0080] As used in this article, "effective amount" is the amount used to treat or prevent an individual's defect, to treat or prevent a disease or medical condition, or more generally, the amount used to reduce symptoms, manage disease progression, or to provide an individual with nutritional, physiological, or medical benefits.

[0081] As used herein, relative terms such as “improvement,” “enhancement,” “enhancement,” etc., refer to the effect of the compositions disclosed herein (i.e., compositions containing the peppermint concentrate disclosed herein) relative to the effect of applying a composition that is otherwise identical but lacks / does not contain the peppermint concentrate disclosed herein during the same time period.

[0082] As used herein, “application” includes another individual providing the mentioned composition to an individual so that the individual can consume the composition, and also includes only the act of the individual consuming the mentioned composition himself.

[0083] As used herein, the term "animal" includes, but is not limited to, mammals, including but not limited to rodents; aquatic mammals; livestock, such as dogs, cats and other pets; farm animals, such as sheep, pigs, cattle and horses; and humans. When using "animal," "mammal," or their plural forms, these terms also apply to any animal capable of having an effect that is manifested or intended to be manifested by the context of the paragraph, such as an animal benefiting from improved mitochondrial calcium input. While the terms "individual" or "subject" are commonly used herein to refer to humans, this disclosure is not limited thereto. Therefore, the terms "individual" or "subject" refer to any animal, mammal, or human who may benefit from the methods and compositions disclosed herein.

[0084] As used herein, the term "pet" means any animal that may benefit from or enjoy the compositions provided in this disclosure. For example, a pet can be a bird, a bovine, a canine, a horse, a feline, a goat, a wolf, a rodent, a sheep, or a pig, but it can also be any suitable animal. The term "companion animal" means a dog or a cat.

[0085] As used herein, "subject" or "individual" refers to a mammal, preferably a human. In the case of humans, the term "elderly" means being at least 60 years old from birth, preferably 63 years or older, more preferably 65 years or older, and most preferably 70 years or older. In the context of humans, the term "middle-aged or elderly" means being at least 45 years old from birth, preferably 50 years or older, more preferably 55 years or older, and includes elderly individuals.

[0086] As used in this article, “frailty” is defined as a clinically identifiable state of increased vulnerability due to a decline in age-related reserves and function across multiple physiological systems, impairing the ability to cope with daily or acute stress. The presence of one or two of these criteria in a pre-frailty stage identifies a high risk of progression to frailty.

[0087] As used herein, the terms "serving" or "unit dosage" are interchangeable and refer to a physically discrete unit suitable as a unit dose for use in human and animal subjects, each unit preferably comprising a predetermined amount of the disclosed peppermint concentrate composition, sufficient to produce the desired effect, in conjunction with a pharmaceutically acceptable diluent, carrier, or mediator. The specifications of the unit dosage form depend on the specific compound used, the desired effect, and the pharmacodynamics associated with each compound in the host. In one embodiment, the unit dosage form may be a predetermined amount of liquid contained in a container such as a bottle.

[0088] As used herein, "oral nutritional supplement" or "ONS" is a composition comprising at least one macronutrient and / or at least one micronutrient, for example in sterile liquid, semi-solid, or powder form, and intended to supplement other nutritional intakes, such as those from food. Non-limiting examples of commercially available ONS products include MERITENE. ® BOOST ® NUTREN ® and SUSTAGEN ® In some embodiments, the ONS can be a beverage in liquid form that can be consumed without further addition of liquid, such as a portion of liquid in a composition.

[0089] As used herein, "incomplete nutrition" preferably refers to a nutritional product that does not contain adequate amounts of macronutrients (proteins, fats, and carbohydrates) or micronutrients, and is insufficient to serve as the sole source of nutrition for the animal to which it is administered. The term "complete nutrition" refers to a product that can serve as the sole source of nutrition for an individual. From a complete nutritional composition, an individual can obtain 100% of its nutritional needs.

[0090] As used herein, “metabolic fatigue” refers to a decline in mitochondrial function in one or more cells (e.g., cells in the liver, kidneys, brain, or skeletal muscle) due to insufficient substrates within one or more cells and / or accumulation of metabolites within muscle fibers that interfere with calcium release or the ability of calcium to stimulate mitochondrial function. Physiological states associated with metabolic fatigue may include muscle fatigue, muscle weakness, lack of energy, weakness, or lack of energy, particularly a lack of bodily energy.

[0091] As used herein, the term "added organic solvent" refers to an organic solvent that is exogenous to the peppermint material and is added in addition to the peppermint material for the preparation of a peppermint material concentrate. The term "added organic solvent" does not include organic solvents that are inherently present in the peppermint material of the peppermint material concentrate.

[0092] As used in this article, the term "vegan" refers to an edible composition that contains no animal products or products of animal origin.

[0093] As used in this article, the term "vegetarian" refers to an edible composition that does not contain meat (including fish).

[0094] As used in this article, the term “biological accessibility” refers to the fraction of the total amount of matter that is theoretically available for absorption.

[0095] As used herein, the term "GAE" refers to gallic acid equivalent. This term is used when quantifying the content of a component against a gallic acid calibration curve. Gallic acid equivalent means that each quantified component is equivalent to one molecule of gallic acid. In other words, 1 mg GAE / g of the quantified component is equivalent to 1 mg / g of the quantified component.

[0096] As used herein, the term "mint" refers to any plant from the genus *Mentha*, preferably any edible plant from the genus *Mentha*. More preferably, mint refers to plants selected from the list consisting of: spearmint (*Mentha spicata*), peppermint (*Mentha × piperita*), or combinations thereof. Most preferably, mint refers to peppermint, especially peppermint.

[0097] As used herein, the terms “blending” and “mixing” are used interchangeably.

[0098] Methods for obtaining peppermint concentrate

[0099] The peppermint concentrate provided in this article can be obtained by a method including the following steps or by that method:

[0100] a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension.

[0101] b) Blend the peppermint material suspension to obtain a peppermint material slurry.

[0102] c) Using physical methods to separate and obtain peppermint concentrate from peppermint slurry.

[0103] d) Optionally, dried peppermint concentrate.

[0104] In one embodiment, the ratio of peppermint material to aqueous liquid in the peppermint material suspension is 1:3 to 1:20, preferably 1:5 to 1:20, more preferably 1:10 to 1:20, and most preferably 1:12 to 1:18.

[0105] Peppermint material contains peppermint plant cells. Similarly, peppermint material suspension contains peppermint plant cells, including whole peppermint plant cells. The peppermint plant cells in the peppermint material suspension are derived from peppermint material.

[0106] The peppermint material suspension is the product obtained in step a). The peppermint material slurry is the product obtained in step b). The peppermint material suspension and the peppermint material slurry are different. Specifically, in the peppermint material slurry, the peppermint plant cells are destroyed and the intracellular substances of the peppermint plant cells are released, while in the peppermint material suspension, the peppermint plant cells are not destroyed, so that the intracellular substances of the peppermint plant cells are not released.

[0107] This aqueous liquid, and therefore the peppermint material suspension, contains no added organic solvents. For example, this aqueous liquid, and therefore the peppermint material suspension, contains no added organic solvents selected from the list consisting of: acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, tert-butanol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxyethane (dimethyl glycol dimethyl ether, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4-dioxane, 1, 2-Dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerol, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, dichloromethane, N-methyl-2-pyrrolidone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tris(hydroxymethyl)methylglycine, tris(hydroxymethyl)aminomethane, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0108] In a preferred embodiment, the aqueous liquid in step a) comprises at least 80% by weight water, more preferably 90% by weight water, and even more preferably at least 95% by weight water. Most preferably, the aqueous liquid in step a) is water.

[0109] In one embodiment, a penetrant may be further added to the peppermint material suspension prior to step b). The penetrant may be selected from a list consisting of glucose, glycerol, sucrose, sorbitol, sodium chloride, potassium chloride, or combinations thereof. Preferably, the penetrant is sucrose. Those skilled in the art can readily determine the amount of penetrant to be added to the peppermint material suspension based on the type of penetrant and the isotonicity of the suspension. The penetrant can be used to adjust the osmotic pressure. This can help maintain the integrity of some plant structures storing bioactive compounds and / or micronutrients and prevent them from breaking down under osmotic pressure, which is also desirable. Furthermore, this allows for further improvement in the stability of bioactive compounds and / or micronutrients.

[0110] In a preferred embodiment, acid may be further added to the peppermint material suspension prior to step b).

[0111] In one embodiment, acid is added to the peppermint material suspension prior to step b) until a pH of 2 to 5.5, preferably 2.5 to 4.5, and most preferably 3 to 4 is reached.

[0112] The acid may be selected from the group consisting of hydrochloric acid, citric acid, malic acid, ascorbic acid, acetic acid, lactic acid, propionic acid, fumaric acid, tartaric acid, phosphoric acid, adipic acid, succinic acid, gluconic acid, or mixtures thereof. Preferably, the acid is selected from the group consisting of malic acid, citric acid, hydrochloric acid, or mixtures thereof. In an even more preferred embodiment, the acid is selected from the group consisting of citric acid, hydrochloric acid, or mixtures thereof. In a most preferred embodiment, the acid is hydrochloric acid. In another most preferred embodiment, the acid is citric acid. The acid may be provided as a pure acid solution, as a dilute acid solution, or as an acidic food ingredient. Examples of acidic food ingredients include citrus juices, such as lemon juice, lime juice, orange juice, tangerine juice, etc.

[0113] The peppermint material suspension may contain 0.01% to 5% by weight of acid.

[0114] When the acid is citric acid, the peppermint material suspension may contain 0.01% to 3% by weight of citric acid, preferably 0.5% to 2.3% by weight of citric acid.

[0115] When the acid is hydrochloric acid, the peppermint material suspension may contain 0.01% to 0.5% hydrochloric acid, preferably 0.02% to 0.54% hydrochloric acid.

[0116] When the acid is malic acid, the peppermint material suspension may contain 0.01% to 3% by weight of malic acid, preferably 0.5% to 2.5% by weight of malic acid.

[0117] The use of acids has a dual effect. In particular, acids lower the pH and chelate micronutrients, especially iron. The decrease in pH and chelation help improve the solubility of micronutrients, especially iron, thus helping to improve the bioavailability of micronutrients, particularly iron, in peppermint concentrate.

[0118] The mint material may comprise any part of the mint plant, such as leaves, stems, flowers, buds, and roots. In one embodiment, the mint material comprises leaves and / or stems from mint. Preferably, the mint material comprises a large amount of leaves. The mint material comprises at least 80% by weight of leaves from mint, more preferably at least 90% by weight of leaves from mint, even more preferably at least 95% by weight of leaves from mint, and even more preferably at least 98% by weight of leaves from mint. The remainder of the mint material may be any part of the mint plant other than leaves from mint disclosed herein. In one embodiment, the remainder of the green plant material consists only of stems from mint.

[0119] In a preferred embodiment, the peppermint material consists only of peppermint leaves.

[0120] Most preferably, the mint material consists only of mint leaves.

[0121] Leaves are preferred because they typically contain a high proportion of bioactive compounds and / or micronutrients, and are edible. Therefore, leaves are a good edible starting material for concentrating large amounts of bioactive compounds and / or micronutrients from peppermint.

[0122] In some embodiments, the peppermint material includes peppermint, spearmint, or combinations thereof. In a preferred embodiment, the peppermint material consists of peppermint, spearmint, or combinations thereof. In a most preferred embodiment, the peppermint material consists of peppermint.

[0123] In one embodiment, the peppermint material is dried peppermint material and / or fresh peppermint material. For example, the peppermint material is dried leaves and / or fresh leaves from peppermint. Advantageously, the peppermint material is dried peppermint material. Dried peppermint material is easier to process on an industrial scale because it has a longer shelf life than fresh peppermint material.

[0124] In one embodiment, when the peppermint material comprises or is dried peppermint plant material, the dried peppermint material can be ground into powder prior to step a). The dried peppermint material can be ground by dry milling. Dry milling can be achieved using any machine equipped with shearing or cutting devices. For example, dry milling can be performed using a hammer mill, stone mill, roller mill, ball mill, jet mill, colloid mill, stirred media mill, bead mill, pin mill, roller grinder, roller fine grinder, impeller mill, cryogenic mill, rod mill, vibratory mill, cutting mill, disc mill, perforated disc mill, micro-cutting mill, or extrusion equipment.

[0125] In one embodiment, the blending step c) can be performed by any type of shearing or mixing device. Examples of mixing devices are: mixers, kitchen mixers, drum mixers, paddle mixers, stirrers, flow impellers, planetary mixers, multi-shaft mixers, Scanima mixers, or Stephan mixers. In one embodiment, blending can be performed in step b) for at least 8 seconds, preferably 8 seconds to 5 minutes, more preferably 1 minute to 3 minutes. In one embodiment, blending can be performed in step b) at a temperature of 4°C to 80°C, preferably 4°C to 25°C, more preferably 10°C to 25°C. The preferred temperature range of 4°C to 25°C is advantageous because it limits the oxidation / chemical degradation of plant organelles that may occur at higher temperatures (e.g., 60°C to 100°C). For example, blending can be performed at room temperature. This step allows the disruption of plant cells and the release of their intracellular substances, including bioactive compounds and / or micronutrients. This helps improve the bioavailability of the bioactive compounds and / or micronutrients in the final concentrate when ingested by humans.

[0126] In one implementation, step c) is performed by filtration and / or centrifugation and / or decantation and / or heat treatment.

[0127] In one implementation, the filtering in step c) can be performed using the same conditions or features as the filtering step c1) provided below in the “Filtering in step c1)” section.

[0128] In one implementation, the heat treatment in step c) can be performed under the same conditions or with the same characteristics as the heat treatment step c2) provided below in the “Step c2) Heat Treatment” section.

[0129] In one implementation, the decantation or centrifugation of step c) can be performed under the same conditions or characteristics as step c3) of the decantation or centrifugation provided below in the “Step c3) Decantation or Centrifugation” section.

[0130] In one implementation, step c) is performed using physical means through the following steps:

[0131] c1) Filter the peppermint slurry to obtain the permeate.

[0132] c2) Optionally, the permeate is heat-treated.

[0133] c3) Centrifuge or decant the permeate to obtain a peppermint concentrate.

[0134] Step c1) Filtering

[0135] As described above, in one embodiment, the method may include step c1) filtering the peppermint material slurry of step b) to obtain a permeate.

[0136] After filtration in step c1), residual material and permeate are obtained. The substance that passes through the filter is called "permeate"; the substance that does not pass through the filter and is recycled is called "residual material". Residual material is removed after step c1), and permeate is recovered and further processed after step c1).

[0137] In a preferred embodiment, step c1) filtration is performed using a filter with a sieve opening of 25 μm to 1000 μm, preferably 25 μm to 500 μm, more preferably 100 μm to 200 μm. This pore size facilitates the separation and concentration of compounds of interest from peppermint (such as bioactive compounds and / or micronutrients), and thus improves their purity, while discarding / reducing undesirable compounds, such as insoluble peppermint compounds. The pore size also reduces the particle size of the peppermint concentrate to a level that makes the concentrate less prone to settling, especially when used in liquid form.

[0138] Step c1) filtration can be performed in one or more steps. In one embodiment, step c1) filtration can be performed in 1 to 10 steps, preferably 1 to 5 steps. When step c1) filtration is performed in several steps, i.e., 2 to 10 steps, preferably 2 to 5 steps, the size of the filter pores decreases in each successive filtration step. In other words, the size of the filter pores used in a predetermined filtration step (e.g., the first filtration step) is larger than the size of the filter pores used in successive and downstream filtration steps (e.g., the second filtration step), and so on.

[0139] In a more preferred embodiment, step c1) filtration is performed in two steps. Specifically, the peppermint material slurry is first filtered with a filter having a sieve opening of 400 to 500 micrometers, preferably 500 micrometers, and then filtered with a filter having a sieve opening of 50 to 200 micrometers, preferably 180 micrometers.

[0140] The filtration process involves several steps, especially two steps, which reduces the tendency for the filter to clog.

[0141] In one embodiment, the sequence of steps a), b), and c1) is repeated at least twice, preferably two to five times, before step c2), and starting from the second sequence of steps a), b), and c1), the peppermint material in step a) is replaced by the permeate obtained in step c1) of the previous sequence of steps a), b), and c1). For clarity, starting from the second sequence of steps a), b), and c1), the permeate, rather than the peppermint material, from the previous sequence of steps a), b), and c1) is suspended in the aqueous liquid of step a) in the continuous sequence of steps a), b), and c1). Therefore, starting from the second sequence of steps a), b), and c1), the suspension of steps a) and b) is not a peppermint material suspension, but a permeate suspension, and the slurry of steps b) and c1) is not a peppermint material slurry, but a permeate slurry. Additionally, permeate is still obtained in step c1) starting from the second sequence of steps a), b), and c1). Furthermore, starting from the second sequence of steps a), b), and c1), residue is also obtained in step c1). The obtained residue can be further processed in the successive sequences of steps a), b), and c1).

[0142] Step c2) Heat treatment

[0143] As described above, in one embodiment, the method may include step c2) optionally heat-treating the permeate obtained in step c1). This heat treatment step allows for an extension of the shelf life of the final peppermint concentrate. In one embodiment, step c2) is not optional.

[0144] In one embodiment, step c2) heat treatment is performed at a temperature of at least 60°C for at least 2 seconds. Preferably, step c2) heat treatment is performed at a temperature of 60°C to 125°C for 2 seconds to 30 minutes. More preferably, step c2) heat treatment is performed at a temperature of 70°C to 85°C for 1 minute to 3 minutes.

[0145] Step c3) Decanting or centrifugation

[0146] As described above, in one embodiment, the method may include step c3) centrifuging or decantation of the permeate to obtain a peppermint concentrate. Preferably, step c3) is a centrifugation step of the permeate. In one embodiment, step c3) centrifugation is performed at 500 g to 10000 g, preferably 1000 g to 5000 g, more preferably 1000 g to 3000 g. In one embodiment, step c3) centrifugation is performed for 2 minutes to 30 minutes, preferably 2 minutes to 20 minutes, more preferably 5 minutes to 15 minutes.

[0147] After centrifugation or decantation, a supernatant and a precipitate are obtained. The precipitate corresponds to the material that forms a sediment at the bottom of the centrifuge / decantation vessel; it is typically a solid or semi-solid (especially a paste), while the supernatant corresponds to the material that floats or lies above the precipitate; it is typically a liquid. The supernatant is discarded. The precipitate is recovered. The precipitate obtained after step c3) corresponds to the peppermint concentrate.

[0148] In one implementation, step c3) involves centrifugation or decantation only once. In other words, the precipitate obtained in step c3) is not further centrifuged or decanted.

[0149] After drying in step d), the peppermint concentrate is not in a semi-solid form (especially a paste form), but in powder form. For example, the drying step can be carried out by spray drying, roller drying, air drying, or freeze drying. In one embodiment, drying in step d) is not optional.

[0150] As an alternative to drying the peppermint concentrate into a powder, the water activity of the peppermint concentrate can be reduced to improve its microbial stability over time. Therefore, in an alternative embodiment, the method may include step d') reducing the water activity of the peppermint concentrate after applying physical means in step c) or centrifugation or decantation in step c3). After reducing the water activity in step d'), the peppermint concentrate has a water activity below 0.85, preferably from 0.5 to 0.85. After this water activity reduction in step d'), the peppermint concentrate is not in powder form. In fact, the peppermint concentrate obtained after step d') is in the same form as the peppermint concentrate obtained after step c) or step c3), i.e., in a semi-solid form (especially a paste form). However, the peppermint concentrate obtained after step d') has a lower water activity than the peppermint concentrate obtained after step c) or step c3). This water activity reduction in step d') can be carried out by evaporating the peppermint concentrate, by drying the peppermint concentrate, or by adding a humectant to the peppermint concentrate. Preferably, the humectant is sucrose. Those skilled in the art can easily determine the amount of humectant to be added to the peppermint concentrate based on the type of humectant and the target water activity. The drying step can be performed by freeze-drying, spray drying, air drying, or roller drying. The evaporation step can be performed using an evaporator.

[0151] In some embodiments, step d') reducing the water activity of the peppermint concentrate and step d) drying the peppermint concentrate can be performed sequentially. In this embodiment, step d') reducing the water activity of the peppermint concentrate occurs before step d) drying the peppermint concentrate.

[0152] In some implementations, particularly when step d) drying is applied, the method does not include evaporation or drying prior to step d) drying.

[0153] In some implementations, particularly when step d) drying is applied, the method does not include any evaporation or drying steps.

[0154] In some implementations, when step d) drying is not applied, the method does not include any evaporation or drying steps.

[0155] In some implementations, step c) is not performed by evaporation or does not involve any use of evaporation equipment, such as a rotary evaporator. Similarly, steps c1, c2, c3, and c4 are not performed by evaporation or do not involve any evaporation or any use of evaporation equipment, such as a rotary evaporator.

[0156] It has been observed that, compared with methods that directly apply drying or evaporation to the permeate as a concentration method without additional physical separation, especially without a centrifugation step, the iron concentration in the final concentrate, expressed in ppm, is significantly increased in the method of the present invention.

[0157] In some embodiments, the method may include a step d") heat-treating the peppermint concentrate after step c) or step c3). This step d") heat treatment may be performed before or after step d'). This step d") heat treatment may be performed at a temperature of at least 60°C for at least 2 seconds. Preferably, the step d") heat treatment is performed at a temperature of 60°C to 125°C for 2 seconds to 30 minutes.

[0158] This method operates to efficiently concentrate bioactive compounds and / or micronutrients, including bioactive compounds and / or micronutrients that guide and selectively regulate mitochondrial Ca2+ homeostasis.

[0159] The obtained concentrate contains a high amount of bioactive compounds and / or micronutrients, including iron. This concentrate guides and selectively regulates mitochondrial Ca2+ homeostasis through its composition of bioactive compounds and / or micronutrients. The concentrate is derived from plant material. Therefore, it is of natural origin and suitable for vegetarian / vegan diets. Furthermore, the concentrate has good sensory properties and, when used in orally delivered / eaten products (e.g., beverages, food products, etc.), produces no sensory defects or very limited sensory defects, particularly limited metallic off-flavors. Additionally, the concentrate is an important source of iron, and the iron in this concentrate has satisfactory bioavailability. Without being bound by theory, the significant amount of iron in this peppermint concentrate may confer immune benefits, particularly contributing to the maintenance of immune function.

[0160] This method is essentially natural. It does not involve the use of added organic solvents, yet still allows for the efficient concentration of bioactive compounds and / or micronutrients (including iron).

[0161] In a preferred embodiment, the method does not involve the use of any added organic solvents. For example, this method does not involve the use of any added organic solvent selected from the list consisting of: acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, 3-butanone, tert-butanol, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, diethylene glycol, diethyl ether, diethylene glycol dimethyl ether, 1,2-dimethoxyethane (dimethyl glycol dimethyl ether, DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,4-dioxane, 1,2-dichloroethane, ethanol, ethyl acetate, ethylene glycol, glycerol, heptane, hexamethylphosphoramide (HMPA), hexane, methanol, dichloromethane, N-methyl-2-pyrrolidone (NMP), nitromethane, naphthalene, pentane, 1-propanol, 2-propanol, pyridine, toluene, triethylamine, tris(hydroxymethyl)methylglycine, tris(hydroxymethyl)aminomethane, tetrahydrofuran, o-xylene, m-xylene, p-xylene, and combinations thereof.

[0162] This method allows for the efficient concentration of micronutrients, such as iron, in the peppermint material. Advantageously, the iron concentration (by weight percentage) in the peppermint concentrate obtained in step c) or c3) is at least twice, preferably two to ten times, higher than the iron concentration in the peppermint material obtained in step a). For example, the iron concentrations in the peppermint concentrate and the peppermint material can be measured according to the methods provided in the examples.

[0163] This method allows for the effective reduction of the ratio of unwanted anti-nutritional factors to micronutrients such as iron, particularly anti-nutritional factors that may reduce or prevent the absorption of micronutrients such as iron in the body.

[0164] Specifically, this method significantly increases the molar ratio (M / M) of iron to oxalic acid. Increasing the molar ratio of iron to oxalic acid to reduce the effect of oxalic acid on iron is advantageous. In fact, oxalic acid may reduce or prevent the absorption of iron in the body. Advantageously, the molar ratio of iron to oxalic acid in the peppermint concentrate obtained in step c) or c3) is at least twice as high, preferably two to ten times higher, than that in the peppermint material obtained in step a). The molar ratio of iron to oxalic acid in the peppermint concentrate is expressed as the dry weight of the peppermint concentrate. The molar ratio of iron to oxalic acid in the peppermint material is expressed as the dry weight of the peppermint material. For example, the molar ratio of iron to oxalic acid in the peppermint concentrate and the peppermint material can be measured according to the method provided in the examples.

[0165] In some embodiments, the method of the present invention significantly increases the molar ratio (M / M) of iron to phytic acid. Increasing the molar ratio of iron to phytic acid to reduce the effect of phytic acid on iron is advantageous. In fact, phytic acid may reduce or prevent the absorption of iron in the body. Advantageously, the molar ratio of iron to phytic acid in the peppermint material concentrate obtained in step c) or c3) is at least twice as high, preferably at least three times as high, and more preferably three to ten times as high as that in the peppermint material in step a). The molar ratio of iron to phytic acid in the peppermint concentrate is expressed as the dry weight of the peppermint concentrate. The molar ratio of iron to phytic acid in the peppermint material is expressed as the dry weight of the peppermint material. For example, the molar ratio of iron to phytic acid in the peppermint concentrate and the peppermint material can be measured according to the method provided in the examples.

[0166] In some embodiments, the method does not include any step of adding enzymes. For example, the method does not include any step of adding protein-degrading enzymes, carbohydrate-degrading enzymes, cellulase-degrading enzymes, oxalate-degrading enzymes, phytate-degrading enzymes, and / or phenolic compound-degrading enzymes.

[0167] Peppermint Concentrate

[0168] In one embodiment, the peppermint concentrate disclosed herein contains at least 500 ppm of iron based on the dry weight of the peppermint concentrate. In a preferred embodiment, the peppermint concentrate contains at least 1000 ppm of iron based on the dry weight of the peppermint concentrate, more preferably at least 1500 ppm of iron. In one embodiment, the peppermint concentrate contains at most 15000 ppm of iron based on the dry weight of the peppermint concentrate, preferably at most 4000 ppm of iron. In one embodiment, the peppermint concentrate is derived from peppermint material. In particular, the peppermint concentrate contains peppermint material. The peppermint material can be the peppermint material provided above in the "Methods for Obtaining Peppermint Concentrate" section. For example, the iron concentration of the peppermint concentrate can be measured according to the methods provided in the examples.

[0169] In some embodiments, the peppermint concentrate disclosed herein has an iron bioavailability of at least 3%, preferably at least 9%, more preferably at least 10%, and even more preferably at least 15%. In some other embodiments, the peppermint concentrate disclosed herein has an iron bioavailability of up to 50%, preferably up to 35%, and more preferably up to 25%. The iron bioavailability of the peppermint concentrate can be measured as provided in the examples.

[0170] In one embodiment, the peppermint concentrate disclosed herein has an absolute amount of bioavailable iron of at least 200 ppm, preferably at least 300 ppm, more preferably at least 350 ppm, and even more preferably at least 500 ppm. In some other embodiments, the peppermint concentrate has an absolute amount of bioavailable iron of up to 1500 ppm, preferably up to 1000 ppm, more preferably up to 800 ppm, even more preferably up to 600 ppm, even more preferably up to 150 ppm, and even more preferably up to 50 ppm. For example, the absolute amount of bioavailable iron in the peppermint concentrate can be measured according to the methods provided in the examples.

[0171] In one embodiment, the peppermint concentrate disclosed herein has a molar ratio (M / M) of iron to oxalic acid of at least 0.3, preferably 0.3 to 3, more preferably 0.4 to 3, even more preferably 0.4 to 1.5, and most preferably 0.4 to 0.8. The molar ratio of iron to oxalic acid is expressed by the dry weight of the peppermint concentrate. For example, the molar ratio of iron to oxalic acid in the peppermint concentrate can be measured according to the methods provided in the examples.

[0172] In some embodiments, the peppermint concentrate disclosed herein contains less than 15,000 ppm, preferably less than 12,000 ppm, and more preferably less than 10,500 ppm of oxalic acid based on the dry weight of the peppermint concentrate. For example, the concentration of oxalic acid in the peppermint concentrate can be measured according to the methods provided in the examples.

[0173] In some embodiments, the peppermint concentrate disclosed herein has a molar ratio (M / M) of iron to phytic acid of at least 5, preferably at least 7, more preferably at least 7.5. In a particular embodiment, the peppermint concentrate disclosed herein has a molar ratio of iron to phytic acid of 5 to 80, preferably 7 to 80, more preferably 7.5 to 80, even more preferably 7.5 to 60, and most preferably 7.5 to 55. The molar ratio of iron to phytic acid is expressed as the dry weight of the peppermint concentrate. For example, the molar ratio of iron to phytic acid in the peppermint concentrate can be measured according to the methods provided in the examples.

[0174] In some embodiments, the peppermint concentrate disclosed herein contains less than 3000 ppm, preferably less than 2000 ppm, and more preferably less than 1900 ppm, based on the dry weight of the peppermint concentrate. For example, the concentration of phytic acid in the peppermint concentrate can be measured according to the methods provided in the examples.

[0175] In one embodiment, the peppermint concentrate disclosed herein is free of any added organic solvents, particularly any added organic solvents listed above in the "Methods for Obtaining Peppermint Concentrates" section.

[0176] In one embodiment, the peppermint concentrate disclosed herein has a pH of 3 to 8.

[0177] In one embodiment, the peppermint concentrate disclosed herein contains 0.01% to 5% by weight of an acid. The acid may be as provided above in the "Methods for Obtaining Peppermint Concentrate" section. Preferably, the acid is selected from the group consisting of ascorbic acid, malic acid, citric acid, hydrochloric acid, or mixtures thereof. In a more preferred embodiment, the acid is selected from the group consisting of malic acid, citric acid, hydrochloric acid, or mixtures thereof. In an even more preferred embodiment, the acid is selected from the group consisting of citric acid, hydrochloric acid, or mixtures thereof. In a most preferred embodiment, the acid is hydrochloric acid. In another most preferred embodiment, the acid is citric acid.

[0178] Acids can be provided as pure acid solutions, as dilute acid solutions, or as acidic food ingredients. Examples of acidic food ingredients include citrus juices, such as lemon juice, lime juice, orange juice, and tangerine juice.

[0179] When the acid is citric acid, the peppermint concentrate disclosed herein may contain 0.01% to 3% by weight, preferably 0.5% to 2.3% by weight, of citric acid.

[0180] When the acid is hydrochloric acid, the peppermint concentrate disclosed herein may contain 0.01% to 0.5% hydrochloric acid, preferably 0.05% to 0.5% hydrochloric acid.

[0181] When the acid is malic acid, the peppermint concentrate disclosed herein may contain 0.01% to 3% by weight of malic acid, preferably 0.5% to 2.5% by weight of malic acid.

[0182] In one embodiment, the peppermint concentrate disclosed herein may contain a penetrant. The penetrant may be one as provided above in the "Methods for Obtaining Peppermint Concentrate" section.

[0183] In one embodiment, the peppermint concentrate disclosed herein may include a humectant. The humectant may be one as provided above in the "Methods for Obtaining Peppermint Concentrate" section.

[0184] In one embodiment, the peppermint concentrate disclosed herein may contain 1% to 50% by weight, preferably 5% to 20% by weight, of total sucrose. The sucrose in the peppermint concentrate may be used as a humectant and / or penetrant. Regardless of whether sucrose is used as a humectant and / or penetrant, the range of total sucrose content provided herein is applicable.

[0185] The peppermint concentrate disclosed herein contains a large number of bioactive compounds and / or micronutrients, including iron. The peppermint concentrate guides and selectively regulates mitochondrial Ca2+ homeostasis through its composition of bioactive compounds and / or micronutrients. The peppermint concentrate is derived from plant material. Therefore, it is of natural origin and suitable for vegetarian / vegan diets.

[0186] In addition, this peppermint concentrate has good sensory properties and does not produce sensory defects or produces very limited sensory defects when used in oral delivery / edible products, especially very limited metallic odor.

[0187] Furthermore, this peppermint concentrate is an important source of iron, and the iron in this concentrate has satisfactory bioavailability. Without being bound by theory, the significant amount of iron in this peppermint concentrate may provide immune benefits, particularly in helping to maintain immune function.

[0188] Composition

[0189] The composition may further comprise one or more additional bioactive compounds. In particular, the composition may also comprise at least one compound selected from the group consisting of: antioxidants, anti-inflammatory compounds, glycosaminoglycans, prebiotics, fiber, probiotics, fatty acids, enzymes, minerals, trace elements, and / or vitamins. The compounds may be derived from natural sources. Therefore, the compounds may be derived from extracts of plants, animals, fish, fungi, algae, or microbial fermentation. Preferably, the compounds are derived from plants, fungi, or algae. Minerals are considered to be derived from natural sources. In a preferred embodiment, the enzyme may be a protease such as trypsin, or an enzyme extract such as, for example, bromelain.

[0190] The effective amount of peppermint concentrate also varies depending on the specific composition, the recipient's age and condition, and the specific disorder or disease being treated. However, in a general embodiment, 0.20 mg to 20 g may be administered to an individual daily, preferably 1 mg to 20 g daily, more preferably 1 mg to 10 mg daily, and even more preferably 1 g to 10 g daily.

[0191] The composition may contain an effective amount of peppermint concentrate. For example, a single serving or dose of the composition may contain an effective amount, while a package may contain one or more servings, or one or more doses. Optionally, the composition may also contain calcium.

[0192] In one particular embodiment, peppermint concentrate, especially an effective amount of peppermint concentrate, may be applied to a composition that also contains calcium.

[0193] The composition may contain food additives selected from the following: acidifiers, thickeners, buffers or reagents for pH adjustment, chelating agents, colorants, emulsifiers, excipients, flavoring agents, minerals, penetrants, pharmaceutically acceptable carriers, preservatives, stabilizers, sugars, sweeteners, texture agents, vitamins, minerals, and combinations thereof.

[0194] Peppermint concentrate, particularly an effective amount of peppermint concentrate, can be administered in any composition suitable for human and / or animal consumption. In a preferred embodiment, it is administered orally or enterally (e.g., via tube feeding). For example, it can be administered to an individual in the form of a beverage, food product, capsule, tablet, powder, or suspension.

[0195] Non-limiting examples of suitable compositions include food compositions, dietary supplements, dietary supplements (e.g., liquid ONS), nutritional compositions, complete nutritional compositions, beverages, pharmaceuticals, oral nutritional supplements, medical foods, nutritional products, foods for special medical purposes (FSMP), powdered nutritional products reconstituted in water or milk before consumption, food additives, pharmaceuticals, pet foods, and combinations thereof.

[0196] Food products according to the invention may include dairy products, such as fermented milk products, like yogurt, buttermilk, etc.; ice cream; condensed milk; milk; dairy cream; flavored milk beverages; whey-based beverages; toppings; coffee creamer; plant-based dairy analogues; chocolate; cheese-based products; soups; sauces; vegetable purees; condiments; puddings; custards; baby food; nutritional formula foods, such as those for complete nutrition, for example, for infants, children, adolescents, adults, the elderly, or the critically ill; such as cereals and cereal bars.

[0197] Beverages (or drinks) may include, for example, milk- or yogurt-based beverages, fermented milk, protein drinks, coffee, tea, energy drinks, soy drinks, water-based beverages, dairy beverage analogs, fruit and / or vegetable beverages, and fruit and / or vegetable juices.

[0198] Peppermint concentrate, in particular an effective amount, may be applied to food products or beverages that also contain components selected from the group consisting of: proteins, carbohydrates, fats, and mixtures thereof.

[0199] In one embodiment, the protein source is preferably purified protein (i.e., isolated from a natural food component from which the protein is produced). The protein content of the composition is preferably 20% to 99% by weight, for example 20% to 90% by weight, for example 30% to 80% by weight, for example 40% to 80% by weight, for example 50% to 80% by weight, for example 40% to 70% by weight.

[0200] Non-limiting examples of proteins or their sources suitable for use in this composition include hydrolyzed, partially hydrolyzed, or unhydrolyzed proteins or protein sources. They may be derived from any known or other suitable source, such as milk (e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice, corn), or vegetable (e.g., soybean, pea) sources. Combinations of protein sources or types may be used. Non-limiting examples of proteins or their sources include whole pea protein, whole pea protein isolates, whole pea protein concentrates, milk protein isolates, milk protein concentrates, casein protein isolates, casein protein concentrates, whey protein concentrates, whey protein isolates, sodium caseinate or calcium caseinate, whole milk, partially or completely skimmed milk, yogurt, soy protein isolates and soy protein concentrates, and combinations thereof. Combinations of protein sources or types may be used. Preferred proteins include pea protein, oat protein, fava bean protein, whey protein, soy protein, and casein. Casein proteins may, for example, comprise sodium caseinate and calcium caseinate.

[0201] The protein source can be provided by individual amino acids, polypeptides containing amino acids, or mixtures thereof. Certain amino acids are beneficial for many muscle growth, muscle maintenance, and / or muscle enhancement therapies, such as L-arginine, L-glutamine, lysine, and branched-chain amino acids (i.e., leucine, isoleucine, and valine; particularly leucine and isoleucine). These specific amino acids can be provided as a protein source, or they can be supplements to a primary protein source. Therefore, the protein source in this composition may include: one or more branched-chain amino acids (leucine, isoleucine, and valine); one or both of L-arginine and L-glutamine; and lysine. In a preferred embodiment, the composition comprises whey protein and / or casein protein, and one or more (or all) of individual amino acids, such as leucine, isoleucine, and L-arginine.

[0202] In one embodiment, the composition further comprises one or more of a medium-chain triglyceride, such as hexanoic acid, caprylic acid, capric acid, and lauric acid. In one embodiment, the composition further comprises a phospholipid, such as phosphatidylcholine.

[0203] The composition may also contain carbohydrate and / or fat sources. Non-limiting examples of suitable fats include low-erucic acid canola oil, corn oil, and high-oleic sunflower oil. Non-limiting examples of suitable carbohydrates include sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, and mixtures thereof. In addition or alternatively, dietary fiber may be added. Dietary fiber passes through the small intestine without being digested by enzymes and functions as a natural filler and laxative. Dietary fiber may be soluble or insoluble, and generally a mixture of both types is preferred. Non-limiting examples of suitable dietary fiber include soybeans, peas, oats, pectin, guar gum, partially hydrolyzed guar gum, gum arabic, fructooligosaccharides, acidic oligosaccharides, galactooligosaccharides, sialyl lactose, and oligosaccharides derived from animal milk. A preferred fiber mixture is a mixture of inulin and shorter-chain fructooligosaccharides. In one embodiment, the fiber content is between 2 and 40 g / L of the composition, for example, between 4 and 10 g / L.

[0204] One or more other minerals besides calcium may be used in the composition. Non-limiting examples of suitable minerals include boron, chromium, copper, iodine, iron, magnesium, manganese, molybdenum, nickel, phosphorus, potassium, selenium, silicon, tin, vanadium, zinc, and combinations thereof.

[0205] One or more other vitamins may also be used in the composition. Non-limiting examples of suitable vitamins include vitamin A, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin or nicotinamide), vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine, pyridoxal, pyridoxine, or pyridoxine hydrochloride), vitamin B7 (biotin), vitamin B9 (folic acid), and vitamin B12 (various cobalamins; typically cyanocobalamin in vitamin supplements), vitamin C, vitamin D, vitamin E, vitamin K, folic acid, biotin), and combinations thereof. "Vitamin" includes such compounds, provitamins, their derivatives, and analogues that are naturally derived from plant and animal foods or synthesized synthetically.

[0206] One or more food-grade emulsifiers, such as diacetyl tartrate of monoglycerides and diglycerides, lecithin, and / or monoglycerides and diglycerides, may be incorporated into the composition. Suitable salts and stabilizers may also be included.

[0207] The compositions disclosed herein can be used for therapeutic administration using any of a variety of formulations. More specifically, the pharmaceutical compositions may contain a suitable pharmaceutically acceptable carrier or diluent and may be formulated into solid, semi-solid, liquid, or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalers, gels, microspheres, and aerosols. Therefore, the composition can be administered in a variety of ways, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal, and intratracheal administration. The active agent may be systemic after administration or localized by the use of topical, intramural, or implantable devices that act to maintain the active dose at the implantation site.

[0208] In pharmaceutical dosage forms, peppermint concentrate can also be used in combination with other pharmaceutically active compounds as appropriate. The following methods and excipients are merely exemplary and in no way limiting.

[0209] For oral formulations, peppermint concentrate can be used alone or in combination with appropriate additives to prepare tablets, powders, granules, or capsules, for example, in combination with common additives such as lactose, mannitol, corn starch, or potato starch; in combination with binders such as crystalline cellulose, cellulose functional derivatives, gum arabic, corn starch, or gelatin; in combination with disintegrants such as corn starch, potato starch, or sodium carboxymethyl cellulose; in combination with lubricants such as talc or magnesium stearate; and, if necessary, in combination with diluents, buffers, wetting agents, preservatives, and flavoring agents.

[0210] In some embodiments, the composition is in the form of a solid powder, powder rod, capsule, or solution.

[0211] The composition, particularly peppermint concentrate, and more particularly an effective amount of peppermint concentrate, may be administered at least one day per week, preferably at least two days per week, more preferably at least three or four days per week (e.g., every other day), most preferably at least five days per week, six days per week, or seven days per week. The duration of administration may be at least one week, preferably at least one month, more preferably at least two months, most preferably at least three months, for example at least four months. In one embodiment, dosing is performed at least daily; for example, the subject may receive one or more doses daily. In some embodiments, administration continues for the remainder of the individual's life. In other embodiments, administration occurs until no detectable symptoms of a medical condition persist. In a specific embodiment, administration occurs until a detectable improvement in at least one symptom occurs, and in other cases, the improvement continues.

[0212] In some embodiments, the composition, particularly peppermint concentrate, and more particularly an effective amount of peppermint concentrate, is administered orally daily for at least one week.

[0213] In one embodiment, the composition is free of milk and / or soy.

[0214] In one embodiment, the composition is vegetarian. In another embodiment, the composition is vegan.

[0215] Treatment

[0216] One aspect of this disclosure is a composition comprising an effective amount of peppermint concentrate for (i) improving physiological states associated with metabolic fatigue in one or more cells of an individual, and / or (ii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells of an individual, and / or (iii) enhancing mitochondrial function of an individual, and / or (iv) treating or preventing calcium deficiency / depletion disorders in an individual.

[0217] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0218] In one embodiment, at least a portion of one or more cells is part of at least one body part selected from the liver, kidney, brain, and skeletal muscle.

[0219] In another implementation, physiological states associated with metabolic fatigue include muscle fatigue, muscle weakness, lack of vitality, weakness or lack of energy, especially lack of physical energy.

[0220] In some implementations, the method includes identifying an individual as having the condition or at risk of having the condition prior to administration.

[0221] Another aspect of this disclosure is a composition comprising peppermint concentrate for use in a therapeutically effective amount to treat or prevent mitochondrial-related diseases or conditions associated with altered mitochondrial function (e.g., reducing their morbidity and / or severity) in individuals in need or at risk. The method comprises orally administering an effective amount of peppermint concentrate to the individual in need or at risk.

[0222] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0223] Unbound by theory, it is believed that various types of stress lead to stress-induced damage to mitochondria, thereby reducing their ability to perform a variety of functions essential for overall cellular function. The methods disclosed herein can be used to treat conditions involving mitochondrial stress-induced damage, which can manifest in any of a variety of ways, including but not limited to mitochondrial diseases.

[0224] Mitochondrial diseases result from inherited or spontaneous mutations in mitochondrial or nuclear DNA that lead to alterations in the function of proteins or RNA molecules that normally reside in mitochondria. However, problems with mitochondrial function may affect only certain tissues due to factors that occur during development and growth that are not yet fully understood. Even when tissue-specific isotypes of mitochondrial proteins are considered, it is difficult to explain the varied patterns of affected organ systems in clinically observed mitochondrial disease syndromes.

[0225] Mitochondrial diseases are caused by defects in mitochondria, specific compartments found in every cell of the body, except for red blood cells. Mitochondria are responsible for producing more than 90% of the energy needed to sustain life and support growth. When they malfunction, less and less energy is produced within the cell. Cell damage and even cell death follow. If this process repeats throughout the body, the entire system begins to malfunction, and the lifespan of people affected is severely compromised. Mitochondrial diseases primarily affect children, but are becoming increasingly common in adults.

[0226] Mitochondrial diseases appear to cause the most severe damage to cells in the brain, heart, liver, skeletal muscle, kidneys, and endocrine and respiratory systems.

[0227] Many symptoms of mitochondrial disorders are nonspecific. These symptoms can also exhibit an intermittent progression with periodic exacerbations. Various manifestations of mitochondrial disorders have been mentioned in review articles on mitochondrial medicine, including paroxysmal migraines, as well as myalgia, gastrointestinal symptoms, tinnitus, depression, chronic fatigue, and diabetes. In patients with mitochondrial disorders, clinical symptoms often occur during periods of high energy demand associated with physiological stressors such as illness, fasting, excessive exercise, and extreme ambient temperatures. Furthermore, psychological stressors frequently trigger symptoms, likely because patients cannot generate sufficient ATP to match their higher brain energy demands.

[0228] Depending on which cells are affected, symptoms can include loss of motor control, muscle weakness and pain, gastrointestinal disturbances and difficulty swallowing, slow growth, heart disease, liver disease, diabetes, respiratory complications, epilepsy, visual / hearing problems, lactic acidosis, developmental delays, and susceptibility to infections.

[0229] Mitochondrial diseases include, but are not limited to: Alpert disease; Barth syndrome; β-oxidation deficiency; carnitine deficiency; carnitine-acyl-carnitine deficiency; chronic progressive extraocular muscle palsy syndrome; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; CPT. I deficiency; CPTII deficiency; creatine deficiency syndrome; cytochrome c oxidase deficiency; type II glutaric aciduria; Kearns-Sayre syndrome; lactic acidosis; LCHAD (long-chain acyl-CoA dehydrogenase deficiency); Leber hereditary optic neuropathy; Leydig disease; lethal infantile cardiomyopathy; Left disease; MAD (medium-chain acyl-CoA dehydrogenase deficiency); mitochondrial cell diseases; mitochondrial DNA depletion; mitochondrial encephalomyopathy, lactic acidosis, and stroke-like symptoms; mitochondrial encephalopathy; mitochondrial myopathy; mitochondrial recessive ataxia syndrome; muscular dystrophy, myoclonic epilepsy, and fragmented red fiber disease; myoneurogenic gastrointestinal encephalopathy. encephalopathy; neuropathy, ataxia, retinitis pigmentosa and ptosis; Pearson syndrome; POLG mutation; pyruvate carboxylase deficiency; pyruvate dehydrogenase deficiency; SCHAD (short chain acyl-CoA dehydrogenase deficiency); and very long chain acyl-CoA dehydrogenase deficiency.

[0230] In one implementation, mitochondrial-related diseases or conditions are selected from the group consisting of: stress (e.g., early stress and / or its effects), physiological aging, obesity, decreased metabolic rate, metabolic syndrome, diabetes, diabetic complications, hyperlipidemia, neurodegenerative diseases, cognitive impairment, stress-induced or stress-related cognitive dysfunction, mood disorders (e.g., stress-induced or stress-related mood disorders), anxiety disorders (e.g., stress-induced or stress-related anxiety disorders), age-related neuronal death or dysfunction (e.g., age-related neuronal death or dysfunction not attributable to a specific neurodegenerative disease), musculoskeletal disorders, frailty, pre-frailty, chronic kidney disease, kidney failure, trauma, infection (e.g., infection occurring in the ICU), cancer, hearing loss, macular degeneration, myopathy, and malnutrition, and combinations thereof.

[0231] Therefore, one aspect of this disclosure is a unit dosage form comprising peppermint concentrate for use in an effective amount to treat or prevent at least one condition selected from the group consisting of: stress (e.g., early stress and / or its effects), physiological aging, obesity, decreased metabolic rate, metabolic syndrome, diabetes, diabetic complications, hyperlipidemia, neurodegenerative diseases, cognitive impairment, stress-induced or stress-related cognitive dysfunction, mood disorders (e.g., stress-induced or stress-related mood disorders), anxiety disorders (e.g., stress-induced or stress-related anxiety disorders), age-related neuronal death or dysfunction (e.g., age-related neuronal death or dysfunction not attributable to a specific neurodegenerative disease), musculoskeletal disorders, frailty, pre-frailty, chronic kidney disease, renal failure, trauma, infection (e.g., infection occurring in the ICU), cancer, hearing loss, macular degeneration, myopathy, and malnutrition, and combinations thereof.

[0232] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0233] Another aspect of this disclosure is a method for treating an individual suffering from at least one condition selected from the group consisting of: stress, obesity, decreased metabolic rate, metabolic syndrome, diabetes, cardiovascular disease, hyperlipidemia, neurodegenerative disease, cognitive impairment, stress-induced or stress-related cognitive dysfunction, mood disorder (e.g., stress-induced or stress-related mood disorder), anxiety disorder (e.g., stress-induced or stress-related anxiety disorder), and age-related neuronal death or dysfunction (e.g., age-related neuronal death or dysfunction not attributable to a specific neurodegenerative disease), trauma, infection (e.g., infection occurring in an ICU), or cancer, said method comprising administering to said individual suffering from said at least one condition a composition comprising an effective amount of peppermint concentrate.

[0234] In one embodiment, the hyperlipidemia being treated or prevented includes hypertriglyceridemia. In one embodiment, the hyperlipidemia being treated or prevented includes elevated free fatty acids. In one embodiment, the age-related neuronal death or dysfunction being treated or prevented is achieved by applying the composition to middle-aged or older individuals, such as elderly individuals.

[0235] The stress being treated or prevented can be early stress, i.e., stress experienced from birth until the age of 5. Early stress has been reported to have significant detrimental effects on cognitive performance, including increased or predisposed levels of psychological parameters such as depression, anxiety, and abnormal risk-taking behavior. Increased levels of attention deficit hyperactivity disorder (ADHD), post-traumatic stress disorder (PTSD), and major depressive disorder have been reported in individuals who have experienced early stress.

[0236] Another aspect of this disclosure is a method for delaying the onset of metabolic decline in healthy middle-aged and older adults, maintaining muscle mass, reducing oxidative stress, maintaining immune function, and / or maintaining cognitive function, said method comprising administering to the healthy middle-aged and older adults a composition comprising an effective amount of peppermint concentrate.

[0237] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0238] Another aspect of this disclosure is a composition comprising peppermint concentrate, used in an effective amount to delay the onset of metabolic decline in healthy middle-aged and older adults and / or maintain muscle mass and / or maintain immune function and / or maintain cognitive function in healthy middle-aged and older adults.

[0239] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0240] Another aspect of this disclosure is a method for enhancing at least one of an individual's mental or muscular performance, the method comprising administering to the individual a composition containing an effective amount of peppermint concentrate.

[0241] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0242] Another aspect of this disclosure is a composition comprising peppermint concentrate, used in an effective amount to enhance at least one of an individual's mental or muscular performance.

[0243] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0244] Another aspect of this disclosure is a method for improving or maintaining the cognitive function of an individual, the method comprising administering to the individual a composition containing an effective amount of peppermint concentrate.

[0245] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0246] In one implementation scheme, cognitive functions are selected from the group consisting of: perception, memory, attention, speech comprehension, speech generation, reading comprehension, image creation, learning, reasoning, and combinations thereof.

[0247] Another aspect of this disclosure is a composition comprising peppermint concentrate for use in an effective amount to improve or maintain an individual's cognitive function.

[0248] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0249] In one implementation scheme, cognitive functions are selected from the group consisting of: perception, memory, attention, speech comprehension, speech generation, reading comprehension, image creation, learning, reasoning, and combinations thereof.

[0250] The compositions disclosed herein can also be used to treat any of a variety of other diseases and conditions in which defective or reduced mitochondrial activity is associated with the pathophysiology of the disease or condition, or in which increased mitochondrial function will produce the desired beneficial effect. Non-limiting examples of such conditions include male infertility associated with decreased sperm motility, macular degeneration, and other age-related and genetic eye disorders and hearing loss (e.g., age-related hearing loss).

[0251] Another aspect of this disclosure is a unit dosage form comprising peppermint concentrate for use in an effective amount for at least one of the following: (i) treating mitochondrial-related diseases or conditions associated with altered mitochondrial function, reducing their incidence or severity, and / or (ii) improving physiological states associated with metabolic fatigue in one or more cells, and / or (iii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells, and (iv) treating or preventing calcium deficiency / depletion disorders, and / or (v) increasing metabolic rate, and / or (vi) improving or maintaining cognitive function, and / or (vii) increasing or maintaining mitochondrial function.

[0252] The peppermint concentrate is the peppermint concentrate disclosed herein, and the peppermint concentrate can be obtained by the methods disclosed herein.

[0253] In one implementation, physiological states associated with metabolic fatigue include muscle fatigue, muscle weakness, lack of energy, weakness or lack of energy, particularly lack of physical energy.

[0254] In one implementation, mitochondrial-related diseases or conditions are selected from the group consisting of: stress (e.g., early stress and / or its effects), physiological aging, obesity, decreased metabolic rate, metabolic syndrome, diabetes, diabetic complications, hyperlipidemia, neurodegenerative diseases, cognitive impairment, stress-induced or stress-related cognitive dysfunction, mood disorders (e.g., stress-induced or stress-related mood disorders), anxiety disorders (e.g., stress-induced or stress-related anxiety disorders), age-related neuronal death or dysfunction (e.g., age-related neuronal death or dysfunction not attributable to a specific neurodegenerative disease), musculoskeletal disorders, frailty, pre-frailty, chronic kidney disease, kidney failure, trauma, infection (e.g., infection occurring in the ICU), cancer, hearing loss, macular degeneration, myopathy, and malnutrition, and combinations thereof.

[0255] In one implementation scheme, cognitive functions are selected from the group consisting of: perception, memory, attention, speech comprehension, speech generation, reading comprehension, image creation, learning, reasoning, and combinations thereof.

[0256] In another implementation, the unit dosage form consists primarily of peppermint concentrate.

[0257] In some implementations of different aspects of this disclosure, the individual may be middle-aged or elderly, elderly, or an ICU patient.

[0258] The above application examples do not require uninterrupted, continuous daily application. Instead, brief interruptions may be allowed during application, such as interruptions of two to four days during the application period. The ideal duration of application of the composition can be determined by those skilled in the art.

[0259] In any of the preferred embodiments of the above aspects, the aforementioned benefits / effects (e.g., enhancing at least one of mental or muscular performance, improving or maintaining cognitive function, etc.) are obtained by enhancing mitochondrial calcium uptake in cells, preferably skeletal muscle cells.

[0260] In any of the preferred embodiments of the foregoing aspects, any effect / benefit on mitochondrial function (e.g., increased mitochondrial energy, increased mitochondrial calcium uptake, enhanced mitochondrial function, etc.) is preferably an effect / benefit on the mitochondria of skeletal muscle cells.

[0261] Those skilled in the art will understand that they are free to combine all features of the invention disclosed herein. In particular, features described for products / compositions of the invention can be combined with treatment methods of the invention, and vice versa. Additionally, features described for different embodiments or aspects of the invention / disclosure can be combined.

[0262] Furthermore, if known equivalents exist for a specific feature, such equivalents should be incorporated as expressly mentioned in this specification. Further advantages and features of the invention will become apparent upon reference to the accompanying drawings and non-limiting embodiments.

[0263] Example

[0264] In all the following embodiments, the peppermint material is peppermint raw material, and the peppermint concentrate is a concentrate obtained by processing peppermint using the methods described in the embodiments.

[0265] Example 1 - Method of concentrating peppermint from dried herbs in the absence of acid according to the present invention

[0266] Dried peppermint (peppermint) material, including leaves and stems, was ground into powder. Peppermint A originated from France, while peppermint B originated from Egypt. The powder was mixed with water at a ratio of 1:15 (w:v) and allowed to hydrate for 5 minutes to form a suspension. The suspension was then blended for 1 minute to obtain a slurry. The obtained slurry was filtered through a filter with a mesh size of 500 μm. The permeate was recovered and subsequently filtered through a filter with a mesh size of 180 μm. The permeate was recovered again, and the obtained permeate was heat-treated to a temperature of 71°C for 2 minutes. After cooling to 4°C, the permeate was centrifuged at 2500 g for 10 minutes. The precipitate was recovered to form a peppermint concentrate. Optionally, the concentrate could be dried.

[0267] Example 2 - Method of concentrating peppermint from dried herbaceous plants in the presence of acid according to the present invention

[0268] Dried peppermint (peppermint) material, including leaves and stems, was ground into powder. The powder (50 g) was mixed with water (700 mL) at a ratio of 1:15 (w:v) to prepare a suspension. Pure citric acid (4.5 g anhydrous citric acid), ascorbic acid (16.8 g), hydrochloric acid (3.6 mL 6M HCl solution), or malic acid (4.7 g) was added to the suspension to achieve a pH of 3.5. After acid addition, the suspension was held for 2 minutes to ensure proper hydration of the powder. The suspension was then blended for 2 minutes to obtain a slurry. The obtained slurry was filtered through a filter with a 500 μm mesh size. The permeate was recovered and subsequently filtered through a filter with a 180 μm mesh size. The permeate was recovered again, and the obtained permeate was heat-treated to a temperature of 71 °C for 2 minutes. After cooling to 4 °C, the permeate was centrifuged at 2500 g for 10 minutes. The precipitate was recovered to form a peppermint acid concentrate. Optionally, the concentrate can be dried.

[0269] Example 3—Iron Quantity

[0270] Materials and methods

[0271] Iron content in peppermint material (i.e., dried peppermint powder obtained after grinding) and peppermint concentrates obtained in Examples 1 or 2 were determined by atomic emission spectrometry using microwave inductively coupled plasma atomic emission spectrometry (MP-AES) 4200 (Agilent, Switzerland). For MP-AES analysis, samples (approximately 100 mg to 400 mg) were mineralized in duplicate using an Xpress microwave bomb digestion system (Mars 6, CEM, USA) with 4 mL of 70% HNO3 ultrapure mass (Sigma-Aldric, St. Louis, MO, USA) and 1 mL of 30% H2O2 (Merck KGaA, Darmstadt, Germany). The mineral solutions were then transferred to 50 mL Falcon tubes and adjusted to 20 mL with Milli-Q water. Iron content was measured at a wavelength of 371 nm using external calibration and multi-element standards. The accuracy of the analysis was examined by analyzing standard reference materials (SRM3233, typical diet; NIST, MD, USA).

[0272] result

[0273] Figure 1 The iron concentrations (based on DW) of peppermint material A (i.e., dried peppermint powder obtained after grinding) and peppermint concentrate A obtained according to the method of Example 1 are shown.

[0274] The concentration method of the present invention achieves a significant increase in iron concentration from 298 ppm in peppermint material to 2973 ppm in peppermint concentrate.

[0275] Example 4 - Quantification of anti-nutritional factors

[0276] Materials and methods :

[0277] Oxalic acid was extracted from the sample with water under mechanical stirring. Oxalic acid was determined by coupling ion chromatography (Dionex ICS-5000 with a Dionex Ion PAC AS16 REFIC Analytical column (250 x 2 mm)) with mass spectrometry (SCIEX Triple Quad 5500 with Selexion).

[0278] Phytate was measured using the Megazyme kit for "Phytate / Total Phosphorus". This kit quantifies free phosphorus and total phosphorus in a sample by colorimetric detection. Total phosphorus is defined as phosphorus derived from phytate and other sources, and is measured after the sample has been treated with phytase followed by alkaline phosphatase. Free phosphorus, on the other hand, is defined as phosphorus derived from sources other than phytate within the sample, and is measured without enzyme treatment from the kit. In short, 1 g of sample was mixed with 20 mL of HCl (0.66 M) and stirred vigorously for 3 hours. The extract (1 mL) was centrifuged at 13000 rpm for 10 minutes, and 0.5 mL of the resulting supernatant was neutralized with 0.5 mL of NaOH solution (0.75 M). The neutralized sample extract (0.05 mL) was mixed with distilled water (0.60 mL), the provided buffer I (0.20 mL), and the phytase suspension (0.02 mL) for the quantification of total phosphorus. To quantify free phosphorus, a control sample was prepared by mixing the sample extract (0.05 mL) with distilled water (0.62 mL) and the provided buffer I (0.20 mL). Both samples were vortexed and incubated at 40 °C for 10 min. Distilled water (0.02 mL) and the provided buffer 3 (0.2 mL) were added to the control, while buffer 3 (0.20 mL) and suspension 4 (ADP, 0.02 mL) were added to the sample for total phosphorus. The sample was vortexed and incubated at 40 °C for 15 min. The reaction was stopped by adding 0.30 mL of trichloroacetic acid (50% w / v). The sample was centrifuged at 13000 rpm for 10 min. The supernatant (1 mL) was used for the colorimetric determination of phosphorus. The sample was mixed with 0.5 mL of the colorimetric reagent. The colorimetric reagent was prepared by mixing 1 part ammonium molybdate solution (5% w / v) with 5 parts ascorbic acid (10% w / v) / sulfuric acid (1 M) solution. After mixing the sample with the colorimetric reagent, incubate it at 40°C for 1 hour, and then read the absorbance at 655 nm.

[0279] The concentration of phosphorus is calculated using the measured absorbance, which is then used to calculate the concentration of PA. The concentrations of free phosphorus and total phosphorus in the reaction are calculated using equation (1).

[0280]

[0281] c 磷 =Phosphorus concentration in the sample [g / 100g]

[0282] Average M = Average value of phosphorus standards [μg / ΔA] 标准物 ], where M = μg of phosphorus in the standard solution (i.e., 0.5-7.5) divided by ΔA 标准物 (i.e. A) 标准物X -A 标准物0 )

[0283] vHCl = Original sample extract volume [mL]

[0284] F = Dilution factor (in this study, F = 1)

[0285] ΔA 样品 =The difference in absorbance of the sample (i.e., A) 样品总 -A 样品游离 )

[0286] w = weight of the original sample material [g]

[0287] v = the sample volume [mL] used in the colorimetric determination step (= 1 mL)

[0288] 10000 = Conversion from μg / g to g / 100g

[0289] The calculated bound phosphorus concentration is converted to PA concentration using equation (2). It is assumed that the amount of bound phosphorus measured is solely from PA.

[0290]

[0291] c PA= Concentration of PA in the sample [g / 100g]

[0292] c 磷 =Concentration of bound phosphorus in the sample [g / 100g]

[0293] 0.282 = Mass fraction of phosphorus in PA

[0294] Then calculate the molar ratio between iron and phytic acid.

[0295] result :

[0296] Table 1 shows the amounts of oxalic acid and phytic acid based on dry weight of peppermint concentrate A obtained according to the method of Example 1.

[0297]

[0298] Figure 2 The molar ratio between the iron and oxalic acid molar concentrations in peppermint material A (i.e., dried peppermint powder obtained after grinding) and peppermint concentrate A obtained according to the method of Example 1 is shown. The concentration method of the present invention increases the iron to oxalic acid ratio from 0.25 to 0.52, indicating that the concentration of iron potentially chelated by oxalic acid in the peppermint concentrate is lower than that in the peppermint material, and therefore potentially more iron is available for absorption.

[0299] Figure 3The diagram shows the molar ratio between iron and phytic acid molar concentrations in two different peppermint materials A and B (i.e., dried peppermint powders obtained by grinding dried peppermint materials A and B, respectively) and in dried peppermint concentrates obtained from peppermint materials A and B according to the method of Example 1. The concentration method of the present invention increases the iron to phytic acid ratio from 2.2 in the peppermint materials to 7.7 in the peppermint concentrate.

[0300] The concentration method of this invention increases the iron to phytic acid ratio from 6.6 in peppermint material B to 44 in peppermint B concentrate. The same trend was observed for different batches of raw materials.

[0301] These results indicate that the concentration of iron potentially chelated by phytic acid in peppermint concentrate is lower than that in peppermint material, thus potentially more iron is available for absorption.

[0302] Example 5 - In vitro digestion to quantify iron bioavailability

[0303] Materials and methods :

[0304] In short, 1 g of peppermint material (i.e., dried peppermint powder obtained after grinding) and peppermint concentrate prepared according to the concentration methods of Examples 1 and 2 were mixed with 10 mL of pH 2 KCl 5 mmol + NaCl 140 mmol. After adjusting the pH to 2, 0.5 mL of pepsin solution (prepared by dissolving 200 mg of pepsin in 10 mL of 0.1 M HCl) was added, and the sample was incubated at 37 °C for 1 hour. After 1 hour, the pH was adjusted to 5.5 with 1 M NaHCO3. The sample volume was adjusted to 15 mL by adding 6.7 KCl 5 mmol + NaCl 140 mmol. Pancreatic enzyme solution (2.5 mL, prepared by adding 87.5 mg of pancreatic enzyme and 525 mg of bile extract to 44 mL of 0.1 M NaHCO3) was added, and the sample was incubated at 37 °C for 2 hours to obtain the in vitro digest.

[0305] Iron content in an aliquot (2.5 g) of intact digest was analyzed by MPAES. The remaining sample was centrifuged at 10000 g for 30 min at 4 °C, and the iron content in the 2.5 g supernatant was analyzed by MPAES. Iron bioavailability was defined as:

[0306]

[0307] Iron bioavailability refers to the fraction of the total amount of iron that is theoretically available for absorption.

[0308] result :

[0309] Figure 4 The diagram illustrates the iron bioavailability of a peppermint concentrate prepared from peppermint material B using water according to the concentration method of Example 1, or a peppermint concentrate prepared from peppermint material B using the concentration method of Example 2 in the presence of an acid (i.e., citric acid, hydrochloric acid, malic acid, or ascorbic acid). Compared to a concentrate prepared using water with the concentration method of the present invention in the absence of an acid (11%) or in the presence of another acid such as ascorbic acid (10%), the use of citric acid during the concentration method of the present invention significantly increases iron bioavailability (24%). Hydrochloric acid has a positive effect on iron bioavailability, but to a lesser extent than citric acid, producing 20% ​​iron bioavailability. Malic acid has a positive effect on iron bioavailability, but to a lesser extent than citric acid, producing 18% iron bioavailability.

[0310] Figure 5 The absolute amounts of bioavailable iron contained in peppermint material B and peppermint concentrate are shown. The peppermint concentrate was prepared from peppermint material B with water according to the concentration method of Example 1, or from peppermint material B with water in the presence of citric acid, hydrochloric acid, malic acid, and ascorbic acid, respectively, according to the concentration method of Example 2. The absolute amount of bioavailable iron was calculated by multiplying the iron content in the sample by a bioavailability value. The peppermint concentrate prepared with hydrochloric acid has a significant advantage over the peppermint material because it contains a much higher amount of bioavailable iron (601 ppm in the peppermint concentrate, compared to 288 ppm in the peppermint material).

[0311] Example 6 - Peppermint concentrate and in vitro digests increase mitochondrial activity via mitochondrial calcium uptake. .

[0312] Materials and methods :

[0313] To test the effect of peppermint concentrate B prepared according to Example 1 or its in vitro digestion on mitochondrial activation in living cells, the increase in mitochondrial calcium was measured in myotubes differentiated from C2C12 cells. An in vitro digestion of peppermint concentrate B was prepared according to the method of Example 5. The in vitro digestion simulated the state of peppermint concentrate B after digestion.

[0314] C2C12 cells were purchased from ATCC. C2C12 cells were seeded at a density of 4500 cells / well in 384-well plates in DMEM high glucose (Gibco) + 10% fetal bovine serum. Myotubes differentiated from C2C12 cells by growing them in DMEM containing 2% horse serum for 7 days.

[0315] Mitochondrial calcium measurements were performed using myotubes infected with an adenovirus (from Sirion Biotech) expressing a mitochondrial-targeting calcium sensor luminescent protein (from a mitochondrial mutant) (Montero et al., 2004). For luminescent protein remodeling, cells or myotubes were incubated at room temperature (22±℃) for 2 hours at 48 hours post-infection in a standard luminescent protein buffer containing 1 μM wild-type coelenterin (145 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 10 mM glucose, and 10 mM Hepes, pH 7.4). Cells were then washed with a modified luminescent protein buffer.

[0316] For sample preparation, peppermint concentrate B or in vitro digests of different concentrations were prepared by diluting peppermint concentrate B or in vitro digests in the above-mentioned standard jellyfish luminescent protein buffer.

[0317] For processing, before measurement, peppermint concentrate B (in different concentrations, such as...) Figure 6 (as shown) or its in vitro digested products (in different concentrations, such as Figure 7 (As shown) Myotube cultures were added directly to standard jellyfish luminescent protein buffer for 2 hours. For the control (ctrl), myotube cultures were incubated with standard jellyfish luminescent protein buffer only, without peppermint concentrate or associated in vitro digests. After incubating myotube cultures with peppermint concentrate or in vitro digests or standard jellyfish luminescent protein buffer (for control) for 2 hours, myotubes were stimulated with 5 mM caffeine to induce mitochondrial calcium rise. Total mitochondrial calcium uptake during stimulation (= total mitochondrial calcium rise) was calculated as the area under the curve. Luminescence was measured on a FLIPR cell imaging reader (molecular device). Luminescence data were calibrated to calcium concentration using the algorithm described above (Alvarez & Montero, 2002). Quantification was performed using a custom module analysis based on Excel (Microsoft) and GhaphPad Prism 7.02 (GraphPad) software.

[0318] result :

[0319] The results are shown in Figure 6 and Figure 7 middle.

[0320] Figure 6 The effect of peppermint concentrate B prepared according to Example 1 at different concentrations on mitochondrial Ca2+ uptake is shown. It can be observed that different concentrations of peppermint concentrate B promote mitochondrial Ca2+ uptake in C2C12-derived myotubules after caffeine stimulation.

[0321] Figure 7The effect of in vitro digests of peppermint concentrate B prepared according to Example 1 at different concentrations on mitochondrial Ca2+ uptake is shown. It can be observed that in vitro digests of peppermint concentrate B at different concentrations promote mitochondrial Ca2+ uptake in C2Cl2-derived myotubes after caffeine stimulation.

Claims

1. A composition containing an effective amount of peppermint concentrate, for (i) improving physiological states associated with metabolic fatigue in one or more cells of an individual, and / or (ii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells of an individual, and / or (iii) enhancing mitochondrial function in an individual, and / or (iv) treating or preventing calcium deficiency / depletion disorders in an individual. The peppermint concentrate can be obtained by a method including the following steps or by said method: a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension. b) Blend the peppermint material suspension to obtain a peppermint material slurry. c) Using physical means to separate and obtain a peppermint concentrate from the peppermint material slurry. d) Optionally, the peppermint concentrate is dried.

2. The composition for the purpose according to claim 1, wherein at least a portion of the one or more cells is part of at least one body part selected from the group consisting of liver, kidney, brain and skeletal muscle.

3. The composition for the use according to any one of the preceding claims, wherein the physiological state associated with metabolic fatigue includes muscle fatigue, muscle weakness, lack of vitality, weakness or lack of energy, particularly lack of physical energy.

4. The composition for the purpose according to any one of the preceding claims, wherein the effective amount of peppermint concentrate is administered orally daily for at least one week.

5. A composition for the purpose according to any one of the preceding claims, wherein the composition further comprises at least one compound selected from the group consisting of: antioxidants, anti-inflammatory compounds, glycosaminoglycans, prebiotics, fiber, probiotics, fatty acids, enzymes, minerals, trace elements and / or vitamins.

6. A composition for the said use according to any one of the preceding claims, wherein the composition is selected from the group consisting of: food compositions, dietary supplements, nutritional compositions, complete nutritional compositions, pharmaceuticals, oral nutritional supplements, medical foods, nutritional products, beverages, powdered nutritional products reconstituted in water or milk before consumption, food additives, foods for special medical purposes (FSMP), pharmaceuticals, pet foods, and combinations thereof.

7. The composition for the purpose according to any one of the preceding claims, wherein the composition is in the form of a solid powder, powder rod, capsule or solution.

8. A composition for the use according to any one of the preceding claims, wherein the effective amount of peppermint concentrate is applied to a food product or beverage, the food product or beverage further comprising a component selected from the group consisting of: proteins, carbohydrates, fats, and mixtures thereof.

9. A composition comprising peppermint concentrate for use in an effective amount to treat, in individuals in need or at risk, mitochondrial-related diseases or conditions associated with altered mitochondrial function, to reduce their morbidity and / or severity, the method comprising orally administering an effective amount of the peppermint concentrate to the individual in need or at risk. The peppermint concentrate can be obtained by a method including the following steps or by said method: a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension. b) Blend the peppermint material suspension to obtain a peppermint material slurry. c) Using physical means to separate and obtain a peppermint concentrate from the peppermint material slurry. d) Optionally, the peppermint concentrate is dried.

10. The composition for the purpose according to claim 9, wherein the mitochondrial-related disease or condition is selected from the group consisting of: stress, physiological aging, obesity, decreased metabolic rate, metabolic syndrome, diabetes, diabetic complications, hyperlipidemia, neurodegenerative diseases, cognitive impairment, stress-induced or stress-related cognitive dysfunction, mood disorders, anxiety disorders, age-related neuronal death or dysfunction, musculoskeletal disorders, frailty, pre-frailty, chronic kidney disease, renal failure, trauma, infection, cancer, hearing loss, macular degeneration, myopathy, and malnutrition, as well as combinations thereof.

11. A composition comprising peppermint concentrate, for use in effective amounts to delay the onset of metabolic decline in healthy middle-aged and older adults, maintain muscle mass and / or muscle function in healthy middle-aged and older adults, maintain immune function in healthy middle-aged and older adults, and / or maintain cognitive function in healthy middle-aged and older adults. The peppermint concentrate can be obtained by a method including the following steps or by said method: a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension. b) Blend the peppermint material suspension to obtain a peppermint material slurry. c) Using physical means to separate and obtain a peppermint concentrate from the peppermint material slurry. d) Optionally, the peppermint concentrate is dried.

12. A composition containing peppermint concentrate, used in an effective amount to enhance at least one of an individual's mental or muscular performance. The peppermint concentrate can be obtained by a method including the following steps or by said method: a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension. b) Blend the peppermint material suspension to obtain a peppermint material slurry. c) Using physical means to separate and obtain a peppermint concentrate from the peppermint material slurry. d) Optionally, the peppermint concentrate is dried.

13. A composition containing peppermint concentrate, used in an effective amount to improve or maintain an individual's cognitive function. The peppermint concentrate can be obtained by a method including the following steps or by said method: a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension. b) Blend the peppermint material suspension to obtain a peppermint material slurry. c) Using physical means to separate and obtain a peppermint concentrate from the peppermint material slurry. d) Optionally, the peppermint concentrate is dried.

14. The composition for the said use according to claim 13, wherein the cognitive function is selected from the group consisting of: perception, memory, attention, speech comprehension, speech generation, reading comprehension, image creation, learning, reasoning, and combinations thereof.

15. The composition for the said use according to any of the preceding claims, wherein the effective amount of peppermint concentrate is applied in a composition that also contains calcium.

16. The composition for the said use according to any of the preceding claims, wherein the individual is a middle-aged or elderly person, an elderly person, or an ICU patient.

17. A unit dosage form containing peppermint concentrate, for use in an effective amount for at least one of the following: (i) treating mitochondrial-related diseases or conditions associated with altered mitochondrial function, reducing their incidence or severity, and / or (ii) improving physiological states associated with metabolic fatigue in one or more cells, and / or (iii) increasing mitochondrial energy and mitochondrial calcium uptake in one or more cells, and / or (iv) treating or preventing calcium deficiency / depletion disorders, and / or (v) increasing metabolic rate, and / or (vi) improving or maintaining cognitive function, and / or (vii) increasing or maintaining mitochondrial function. The peppermint concentrate can be obtained by a method including the following steps or by said method: a) Suspend the peppermint material in an aqueous liquid to form a peppermint material suspension. b) Blend the peppermint material suspension to obtain a peppermint material slurry. c) Using physical means to separate and obtain a peppermint concentrate from the peppermint material slurry. d) Optionally, the peppermint concentrate is dried.

18. The unit dosage form of claim 17, wherein the physiological state associated with metabolic fatigue includes muscle fatigue, muscle weakness, lack of vitality, weakness or lack of energy, particularly lack of bodily energy.

19. The composition for the said use according to any one of claims 1 to 16 or the unit dosage form according to any one of claims 17 or 18, wherein the peppermint concentrate is a peppermint concentrate, and wherein the peppermint material comprises or is composed of peppermint material.

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