Description:FIELD OF INVENTION

[0001] The present invention is directed to a method for preparation of ferula-based formulation, and more particularly to preparation of a formulation from Ferula sp. Extracts with hypoglycemic properties.

BACKGROUND OF INVENTION

[0002] The use of plant extracts and derivatives for healing and prevention has been extensively documented in traditional and folk medicine literature. Nutraceutical are foods or bioactive ingredients in foods that safeguard or enhance health, whether they are delivered as unprocessed agricultural products, processed foods, dietary supplements, extracts, beverages, or other goods. They are produced at the nexus of the food and pharmaceutical industries. In other words, the entire concept is based on disease preventing/treating phytonutrients found in foodstuffs in combating diseases.
[0003] There are over 60 species of the genus Ferula, which is a member of the Umbeliferacaea family, with the majority of them found on the continents of Central Asia, Europe, and North Africa. They are herbaceous, monoecious, perennial plants that can reach heights of 1-4 m and have thick, hollow stems that resemble succulents. They have huge schizogeneous channels and lysigenous chambers filled with milky fluids. After 4-5 years of growth, the massive tap root is chopped off near to the crown after its diameter at the crown reaches 10-15 cm to collect the exudes.
[0004] A few recent reports document that oleo-gum resin from F. asafoetida possess antidiabetic activity. The administration of aqueous asafoetida extract at a dose of 50 mg/kg for 4 weeks demonstrated hypoglycemic activity in streptozotocin-diabetic rats during the second week and at the end of the four-week treatment period (Akhlaghi, 2012). F. asafoetida aqueous extracts treatment led to a significant improvement in hyperglycemia, and increased percentage of body weight in alloxan-induced adult male albino diabetic rats (Helal, Mostafa, MhMood, & Kahwash, 2005). However, no clinical evaluations have so far been reported on the therapeutic efficacy of asafoetida gum against diabetics.
[0005] The non-volatile portion of Ferula resin, which is primarily made up of secondary metabolites, has received most of the attention from major research studies. The majority of the processes discussed in the prior art involve a solvent-based extraction, which excludes the majority of the therapeutic compounds that confer multiple health benefits. Rarely are the studies on therapeutic potential of the volatile fraction or other Ferula oleo-gum components. Furthermore, there is a knowledge gap about the biological activities of other Ferula species besides F. asafoetida that needs to be filled. Besides that, there is no reliable method for producing Ferula extracts with the greatest therapeutic potential.
[0006] In general, a crude extract from a natural habit prepared in a predetermined ratio solely based on weight will not fully satisfy the therapeutic need due to inconsistencies in the natural yield caused by geographical location and seasonality. In the background of foregoing limitations, food composition(s) is desired that promote health, prevent disease, and provide the best nutrients for enhancing the general health of diabetics while also assisting in blood sugar control. Getting an appropriate composition and the right carrier to the target site is the biggest obstacle in therapeutic agent delivery.
[0007]. In the background of foregoing limitations, there exists a need for a low energy consuming, cost effective and commercially viable technology that can enable preparation of ferula-based formulation to eliminate aforementioned health issues. .
OBJECT OF THE INVENTION

[0008] The primary objective of the present disclosure is to provide a novel synergistic food composition with numerous health benefits that contains oleo-gum resin or extracts from a variety of Ferula sp.
[0009] In one other objective of the present disclosure, the method for preparing the novel synergistic food composition is disclosed that can be used to produce a beverage having multiple health benefits.
[0010] Another objective of the present disclosure is to provide a food composition(s) based on various species of Ferula that promote health, prevent disease, and provide the best nutrients for enhancing the general health of diabetics while also assisting in blood sugar control.
[0011] In yet another objective of the present disclosure, the food composition containing Ferula extracts contains various potential carriers that could deliver the primary formulation with the major components intact while maintaining its efficacy.
SUMMARY OF THE INVENTION
[0012] In accordance with one significant aspect of present disclosure, a ferula-based formulation consisting of ferula extracts is disclosed. The ferula extracts here is prepared from a variety of Ferula sp., including Ferula asafoetida, Ferula alliacea, Ferula rubricaulis, Ferula foetida, Ferula foetidissima, Ferula jaeschkeana, Ferula fukanensis, Ferula sinkiangensis and Ferula narthex, either alone or as a combination of two or more species.

[0013] In another aspect of the present disclosure, the ferula extracts are made utilizing ferula resin or a component of the plant, such as leaves, rhizomes, shoots, stems, or roots, or any combination thereof.

[0014] In another aspect, a method for preparation of ferula-based formulation is disclosed, wherein the method comprises a homogeneous mixture of ingredients, and wherein the Ferula sp. extract is at a level of 0.1- 27% w/w with a particle size less than 500 µm, plant gums are at a level of 50- 75% w/w with a particle size less than 500 µm, edible oil at 0.1- 20% v/w, edible starch at a level of 30- 50% w/w with a particle size less than 500 µm, and wherein the homogenous mixture of ingredients is blended for 10-30 minutes, and aged for 12-72 hours in presence of air or nitrogen at temperature ranging between 22- 40°C.

[0015] In yet another aspect, a method for preparing flavored beverage from a ferula-based formulation is disclosed, wherein the method comprises steps of adding starter culture at a level of 0.5–5% to pasteurized milk that has been cooled to 40–50°C, incubating the mixture at 25–45°C for 4–8 hours, and churning coagulated product with a solvent (pH: 6–8) at a ratio of 1:1 until a homogeneous suspension of pH 3.5–5 is obtained, and to which a composition of ferula extract comprising of a whole resin or solvent based extract of resin or a solvent based plant component extract is added at a level of 0.1-5% w/v.

BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 shows process flow chart for preparing ferula based formulation, in accordance with one preferred embodiment of present disclosure.
DETAILED DESCRIPTION OF THE EMBODIMENTS

[0017] Before the present working principle of preparation of formulation from genus Ferula is described, it is to be understood that this disclosure is not limited to the particular means and mode for achieving so, as described, since it may vary within the specification indicated. It is also to be understood that the terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of the present invention, which will be limited only by the appended claims. The words "comprising," "having," "containing," and "including," and other forms thereof, are intended to be equivalent in meaning and be open ended in that an item or items following any one of these words is not meant to be an exhaustive listing of such item or items, or meant to be limited to only the listed item or items. The disclosed embodiments are merely exemplary methods of the invention, which may be embodied in various forms.
[0018] The present disclosure relates to methods and compositions of formulated extracts from the genus Ferula, specifically compositions for oral consumption that contain sulfur volatiles, ferulic acid, and coumarins, as well as their esters, polyacetylenes, and polysaccharides. Furthermore, the present invention relates to how these compositions are used. The present disclosure provides a novel synergistic composition of oleo-gum resin derived from two or more Ferula species for promoting health benefits by lowering blood sugar levels, improving dyslipidemia and/or gastritis, and/or preventing constipation. Further, the present disclosure provides steps for processing extracts from Ferula asafoetida and related species to create compositions with predetermined chemical compound profiles or ratios to satisfy specific requirements for finished goods.

[0019] For the purposes of present disclosure, the term "therapeutically effective" or "effective amount" as used herein refers to a material or amount of material that is effective in preventing, alleviating, or ameliorating one or more symptoms of a disease or medical condition, and/or extending the survival of the subject being treated. In this context, a "combination" is any association between or among two or more items. The combination may consist of two or more distinct items, such as two compositions or two collections. It may be a combination of those things, such as a single mixture of the two or more things, or any variation thereof. The term "composition" refers to any mixture in this context. It could be a solution, a suspension, a liquid, a powder, a paste, aqueous, non-aqueous, or any combination of the above. Also, an extract, as defined herein, is a solution or a slurry containing compound(s), usually in concentrated form, obtained by treating a solid material (such as plant material) with a solvent to remove desired compounds or components.

[0020] In accordance with one exemplary embodiment, a method for preparation of formulation from genus Ferula is disclosed. Firstly, a method for tapping Ferula resin is described. Accordingly, the stem is cut close to the crown of the root, before the flowering begins, in order to collect the oleo-gum-resin. However, traditional cutting methods destroy crown buds, resulting in plant mortality. As a result, suitable tapping technologies for high production of oleo-gum resin on a sustainable basis are required. For the purpose of tapping the root exudates, a lateral incision is performed on the root with a diameter greater than 7 cm, preferably 12 cm.

[0021] The process of collection of oleo-gum-resin and making incision is repeated. The number of incisions is limited to 15 times for achieving the highest production of the therapeutically active composition, however repeated incision, and collection (scrapped off) of oozes from the surface representing oleo-gum resin are done at frequent intervals. To prevent oxidation and evaporation of key volatiles, the roots are covered quickly after each incision. Plant parts such roots, stems, leaves, flower buds, seeds, and rhizomes are harvested from Ferula sp. plants that are at least one year old. For the composition, any of the aforementioned plant parts—fresh or dried, alone or in combination—are further ground.

[0022] In accordance with another exemplary embodiment of present disclosure, composition for preparation of Ferula extract is disclosed. Individual solvent-based extracts of the resin or plant material produced as discussed in aforementioned paragraphs, and can be accomplished using methods known to those skilled in the art, such as Soxhlet extraction, maceration, liquid-liquid extraction, microwave-assisted extraction, ultrasound-assisted extraction, percolation, digestion, infusion, and supercritical fluid extraction, and others.

[0023] Additionally, distillation techniques like hydro distillation, steam distillation, and others can be used to separate the principal components. Solvents such as hexane, dichloromethane, methanol, ethanol, acetonitrile, water, chloroform, ethyl acetate, Supercritical carbon dioxide, and others can be used alone or in combination. Moreover, the extraction process can be accelerated up by employing solutes including salts, polyethylene glycols and others.

[0024] In one working embodiment, the oleo-gum resin and/or plant material of the Ferula species listed in Table 1 are blended in the appropriate range in a mechanically agitated vessel. Alternatively, solvent-based resin extracts and/or solvent-based plant material extracts, or both, are mixed in the suitable range given in Table 1. Another technique of production involves combining plant material, oleogum, and solvent extracts of resin and/or plant material in the appropriate range as described in Table 1. Further, a saturated extract is made by adding a hydrophilic solvent with a pH of 5.8-8 to the oleo-gum resin at a solvent to resin ratio ranging from 0.5:1 to 20:1.

[0025] The pH is adjusted using any available acid or base. Bases such as sodium carbonate, ammonium hydroxide, potassium hydroxide, sodium hydroxide, and others can be used. The hydrophilic solvent is added intermittently, and intermittent mixing is performed for 10-30 minutes to form a homogeneous mixture free of undesirable exothermic reactions, as heat-generating chemical reactions have the potential to produce inconsistent or poor-quality end products. As a result, the temperature is maintained between 25-40°C in complete darkness and the presence of air. The result slurry or suspension is passed through a sieve to remove extraneous matter. This composition will henceforth be denoted as Composition or C1.

Table 1 Ferula species used to make Composition-1 or C1
Ferula species Range used in the composition
Ferula asafoetida > 99 % w/w
Ferula foetida 0.1-99% w/w
Ferula alliacea 0.02-98 % w/w
Ferula foetidissima 0.3-99.3% w/w
Ferula rubricaulis 0.02-99.7% w/w
Ferula narthex 0.1-99.1 % w/w
Ferula jaeschkeana 0.02-98.9 % w/w
Ferula fukanensis 0.02-97% w/w
Ferula sinkiangensis 0.25-99.5% w/w

[0026] In next working embodiment, a method for preparation of flour is discussed. The flour used in the formulation includes either cereal flour, legume flour or combination of two. Cereal flour could be preferably wheat, refined wheat flour, corn flour, rice flour, oats flour. Black beans, black gram, green gram, garbanzo beans (chickpeas), kidney beans, soybeans, green pea, white lentil, and lupin are some examples of legumes from which flour can be made. Alternatively, flour made from starches found in tubers such as taro, potato, arrowroot, cassava, and canna can be used.

[0027] The flour is obtained by milling the ingredients without generation of heat and roughly sieving it on a 500 µm sieve with less than 5% retention followed by a 300 µm sieve with 20-50% retention. The fiber content of this coarse fraction must be at least 1% to 50%, preferably 2-10%. Further processing is carried out on flour with a moisture content of less than 15%, preferably less than 10%. Dehydration or drying can be accomplished using methods known to those skilled in the art, such as freeze drying and spray drying. Food drier or Food dehydrator can be used where the flour is spread out on a heated tray to a uniform thickness of about 10 mm, preferably less than 6mm. The heated tray is then placed in a drier at a constant temperature of no more than 120 °C. Every 15 minutes, for about 1-2 minutes, hot air is blown into the system. The procedure takes about 60-90 minutes to complete.

[0028] In next working embodiment, preparation of emulsifying agent is disclosed. Ferula based formulation further comprises of plant gums such as gum Arabic or synthetic emulsifiers and stabilizers preferably containing polysaccharides and glycoproteins which may act as prebiotics. Preferably, gum Arabic is used due to its excellent emulsifying properties and low viscosity even at a higher concentration. The gum Arabic, preferentially, with a moisture content less than 12%, more preferably a moisture content of 10% by weight is taken for processing. The gum Arabic is spread out on a heated tray to a uniform thickness of about 10 mm, preferably less than 6mm. The heated tray is then placed in a drier at a constant temperature of no more than 120 °C. Every 15 minutes, for about 1-2 minutes, hot air is blown into the system. The process takes between 60 and 90 minutes to finish, and the finished product has a moisture content of 4-7%.

[0029] In accordance with one significant embodiment, method for preparation of Ferula based composition (F1) is disclosed. A food composition comprising a homogeneous mixture of the ingredients as mentioned in aforementioned paragraphs is prepared. A method for making the said formulation (F1) is to blend components, flour obtained in above paragraphs and emulsifying agent, preferably gum Arabic obtained in accordance with above method, in a blender at a temperature of 20- 40°C for about 5-20 minutes until a homogeneous mixture is obtained. The ratio of Flour and gum Arabic can be ranging from 0.5: 2 to 2: 2. Furthermore, C1, obtained as explained above, is then added to the aforementioned mixture, blended for 5-30 minutes at a temperature not exceeding 40°C until a homogeneous mixture with a particle sizes less than 500 µm is obtained.

[0030] The resulting slurry or paste is immediately cooled by spreading it in a tray precooled at 15 °C and allowing it to cool for about 30-45 minutes at a temperature not exceeding 30 °C. The contents are then transferred to airtight containers and allowed to rest for 12-72 hours at a temperature of 20-35°C, preferably at 25°C, to prevent undesirable oxidation. The finished product that constitutes about 0.1- 27% w/w of Ferula sp. extract is either ground into powder or mounded into desired shapes. As a result, the final formulation (F1) can be offered as powder either compressed or loose, granules, capsules, lumps, or cake. The final composition consists of carbohydrates 60-80%, moisture 10-20%, protein 4-12%, fat 0.5-2%, vitamin C 0.001-0.0035%, ash content 1-15%, alcohol extractable matter 4-8%, and volatile oil 0.2-5%.

[0031] The above method steps are followed by addition of oil-based flavoring agent. The oils or fats are not particularly limited but usually mustard oil, sunflower, groundnut, safflower oil, sesame oil is preferred. The usage of oil or fats is optional in F1 obtained in above paragraphs is to further stabilize the emulsion and prevent the evaporation of components. Numerous uses are possible for the food formulation F1 obtained above that is explained below.

[0032] In certain embodiments, the present invention relates to a cooked food comprising of the aforementioned composition. In another embodiments, the food composition F1 can be used to make flavored drinks such as fermented milk, curd, yogurt, buttermilk, and lassi, wherein the milk can be derived from mammals other than cow, buffalo, goat, camel, and sheep.

[0033] In other embodiments, pasteurized milk brought to a temperature of 40-50 °C is mixed with F1 at a concentration of 0.1-10% w/v and thoroughly mixed at an rpm of 50-200 for about 10-20 minutes. A starter culture at a concentration of 0.5-5% is added to this, and the mixture is incubated at 25-45°C for 4-8 hours with starter cultures from the genera Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, and others. The resulting substance is then mixed with an extracting medium (pH: 6-8) in a 1:1 ratio and blended until a homogeneous suspension of pH 3-4.5 is obtained. This process primarily improves texture, aroma, flavor, and health effects.

[0034] Addition of salt-based or other plant extracts as a flavoring agent or preservative can be added to above preparation in one alternate embodiment. In other embodiments, the food composition F1 as obtained above can optionally include a flavoring agent or preservative to impart or maintain a specific flavor over the composition's shelf life. Additionally, a person skilled in the art may use any flavoring agent that can provide a particular flavor profile for the food composition's intended end use. Some of examples include, table salt, black-salt, pink-salt, spices such as pepper, cumin, mustard, ginger, chilly, sugar, herbs such as Coriandrum sp., Murraya koenigii and Mentha sp.

[0035] Addition of nutritionally fortifying agents is now explained for purposes of present disclosure. In other embodiments, the food formulation F1 can optionally include minerals such as calcium, phosphorus, magnesium, iron, zinc, iodine, selenium, copper, manganese, chromium, molybdenum, and chloride, either singly or in combination. Other embodiments allow for the optional inclusion of vitamins such as A, D, E, K, C, thiamin, riboflavin, niacin, vitamin B6, folate, vitamin B12, biotin, and pantothenic acid, either singly or in combination in the food composition F1.

[0036] The food composition F1 when ingested alone or in combination with other preparations may produce health advantages by decreasing blood sugar levels, improving dyslipidemia and/or gastritis, and/or preventing constipation. In certain embodiments, the present invention relates to a cooked food comprising one or more of the aforementioned compositions. The compositions of the present invention can be administered in a dosage range of 50 mg to 2000 mg per dose in any of the mentioned dosage forms. The recommended dose of administration is twice a day. In another aspect, the invention discloses a method of treatment, comprising the step of administering to a subject in need thereof with an effective amount of a composition as described herein for the treatment, prevention, or delay of progression of a disorder as described herein.

[0037] In one exemplary embodiment, the preparation of formulation is explained. Accordingly, a 4-5-year-old Ferula species (listed in Table 2) is repeatedly incised, and the oleo-gum is harvested. A lateral incision is performed on the root with a diameter of about 10 cm and gum from only the first seven incisions are collected for the plant species 1-4 listed in Table 2. For the plant species 5-9 listed in Table 2, gum from only the first five incisions are collected. To collect the oleo-gum with the least amount of light exposure, sufficient care is taken. The gum is checked to see if there are any stones or plant debris. The appropriate amount of oleo gum, as specified in Table 2, is then blended with 20 ml of solvent at 25 °C in complete darkness to create a homogenous slurry for 14 minutes. Then, an additional 40 ml of solvent is added, and the mixture is blended at 100 rpm for about 10 minutes.

[0038] Finally, 40 ml of solvent is added and blended for 10 minutes at 50 rpm. Reverse osmosis treated water with a Total dissolved solids not more than 50 ppm and a pH of 7.3 is used as the solvent in this case. To get rid of extraneous material, this blended suspension is then put through a 400µm sieve to obtain Composition 1 or C1 (Table 2). The following ingredients are added to 5 g of prepared F1: 35 g of refined wheat flour, 75 g of gum Arabic, both previously air-dried at 100 °C with final moisture contents of 12 and 6%. The ingredients are thoroughly blended for 25 minutes at 30°C until a homogeneous mixture is obtained. The resulting slurry or paste is immediately cooled by spreading it in a tray precooled at 15 °C and allowing it to cool for about 45 minutes at 22 °C. The contents are then transferred to airtight containers and allowed to rest for 36 hours at 22 °C. Followed by the incubation time, the contents are finally ground in a 400µm sieve to retain about 30% of the particles. The process flow chart for the same is illustrated in Figure 1.

Table 3 Ferula species used to make Composition 1 or C1 (
S.No Ferula species Range used in the composition (%) (w/w)
1. Ferula asafoetida 18.6
2. Ferula foetida 12.1
3. Ferula alliacea 8.3
4. Ferula foetidissima 13.3
5. Ferula rubricaulis 6.2
6. Ferula narthex 14.7
7. Ferula jaeschkeana 9.1
8. Ferula fukanensis 2.2
9. Ferula sinkiangensis 15.5

[0039] Table 3 below depicts nutritional composition of above formulation F1:
Parameter Composition (%)
carbohydrates 75
Protein 5
Fat 1.3
alcohol extractable matter 5.90
volatile oil 1.3
vitamin C 0.001

[0040] In next working embodiment, method of preparation of formulation is explained. Solvent extracts of Ferula sp. are prepared as follows: 500 g of Dried rhizomes of 4-year-old Ferula sp. are mixed as given in Table 4 and collected in glass thimble of a Soxhlet extractor. In a Soxhlet extractor, a sequential extraction is performed using petroleum ether, ethyl acetate, acetone, and ethanol, with three cycles of each solvent. The solvent is completely evaporated in a rotary evaporator, and the residue is lyophilized for 24 hours to obtain the dry flaky material, which is referred to as the non-volatile fraction.

[0041] Additionally, the oleo-gum is collected by making a lateral incision on the root with a diameter of about 12 cm and gum from only the first seven incisions are collected. Further, essential oils are isolated from oleo gum by hydro distillation (Clevenger apparatus). About 100 g of the Ferula gum are measured out in the proportions shown in the, transferred to the Clevenger apparatus' round bottom flask with 500 ml of water, and the distillation is started at 60 °C for 2 h, then raised up to 80 °C for 2 h, and finally 100 °C for 1 h. The non-volatile fraction is combined with the essential oil following the addition of 4 ml of water, and the combination is blended at 100 rpm for approximately 10 minutes to produce C1.

[0042] The solvent in this example is reverse osmosis treated water with a total dissolved solids concentration of less than 50 ppm and a pH of 6. To 6 g of prepared C1, the following components are added: 13 g of wheat flour, 16 g of soya flour, and 65 g of gum Arabic, previously air-dried at 100 °C with final moisture percentages of 12, 11, and 6%, respectively. The ingredients are thoroughly blended with 2 ml of sesame oil for 25 minutes at 30°C until a homogeneous mixture is obtained. The resultant slurry or paste is immediately cooled by spreading it in a tray precooled at 15 °C and allowing it to cool for around 45 minutes at 20 °C. The contents are then transferred to airtight containers and allowed to rest for 72 hours at 22 °C. After incubation, the contents are crushed in a 400 µm sieve to retain roughly 30% of the particles.

Table 4 Ferula species used to make Composition 1 or C1
Ferula species Range used in the composition (%) (w/w)
Ferula asafetida 48.1
Ferula foetidissima 20
Ferula foetida 10.3
Ferula rubricaulis 6.2
Ferula narthex 3.8
Ferula jaeschkeana 8.7
Ferula fukanensis 1.9
Ferula sinkiangensis 1.0

Table 5 Ferula species used to make Composition 1 or C1
Ferula species Range used in the composition (%) (w/w)
Ferula asafoetida 38.3
Ferula foetida 25.2
Ferula foetidissima 20.7
Ferula rubricaulis 15.8

[0043] In next exemplary embodiment, preparation of a flavored beverage comprising formulation F1 is described. One liter of pasteurized milk is reduced to 45°C and mixed with 2% v/v of Bifidobacterium and 1% v/v of Lactobacillus. This mixture is vortexed for approximately 5 minutes after which 13.34 g of Formulation and 1L of pasteurized milk are added. The blend is then incubated for a further 6 hours at 37°C. A coagulated product is produced and then 6.6 g of sodium chloride and 2L of RO water are added to a churn at a pH of 9. Fresh Mentha leaves are crushed to a coarse paste and added to the beverage in an amount of around 1g. Within two hours, the beverage is consumed after being chilled to 10°C.

[0044] In another exemplary embodiment, analysis of compounds in Formulation F1 is demonstrated. The volatile constituents of the formulation F1 are investigated. Around 100g of the formulated product (F1) is placed in a hydro distillation apparatus, and the temperature is progressively increased from 60 °C to 100 °C at a rate of 10 °C per hour for 5 hours. The oil is further diluted with hexane and analyzed using a High-Resolution Gas Chromatography-Orbitrap Mass Spectrometry (GC-Orbitrap-MS). One microliter of the standard or sample extract is injected into the Q Extractive GC-Orbitrap MS system equipped with a TriPlus™ RSH™ Autosampler (Thermo Scientific) in split mode with a split ratio of 5.

[0045] The inlet temperature is set at 200 °C. The TRACE 1300 series GC operated in constant flow mode at 1.5 mL min-1 with helium as a carrier gas, using the following oven temperature programs: the initial temperature 70 °C is maintained for 2 min, raised to 150 °C at a rate of 5 °C/min, maintained for 3 min, raised to 200 °C at a rate of 10 °C/min held for 3 min, raised to 280 °C at a rate of 10 °C/min and maintained for 10 min. The transfer line and source temperatures are 280 and 250 °C, respectively. The EI ion source operated at 70 eV. Data is recorded in full scan profile mode at 50–600 m/z with a Fourier transform resolution of 60,000.

[0046] For non-volatile analysis, 1 g of the F1 is homogenized with 2 ml of acetonitrile and sonicated for 5 h in a cooling ultrasonic bath set at 10 °C and under complete darkness. The acetonitrile layer is then separated from the pellet by centrifuging the sample at 4000 rpm at 4°C. The pellet is re-suspended in 2.5 ml of water and 2.5 ml of 6N sodium hydroxide. The samples are then sonicated in an ultrasonic bath at 20°C for 45 min. The pH is then decreased to 2.0 with diluted sulphuric acid to allow the hemicellulose to precipitate.

[0047] After centrifuging the samples at 5000 rpm for 10 minutes, the supernatant is transferred to a separating funnel, and an equivalent volume of ethyl acetate is added to perform a liquid-liquid extraction. After phase separation, the organic layer is recovered. The procedure is carried out five times. A rotary evaporator is used to evaporate the organic layer. The resulting residue is then diluted in 2 mL of methanol before being analyzed using LC-MS and/or RP-HPLC using a C18 column.

[0048] In order to test the efficacy of product, a clinical evaluation is performed on the final compound that has been prepared. About, 150 ml of freshly set curd is mixed with 150 ml of drinking water and made into a homogenous suspension using a hand blender. To this, 1 g of the investigational formulation (F1) and 0.5 of table salt is added and the resulting beverage is consumed immediately. An open label, prospective, randomized, two-arm, cohort-based clinical study is designed to evaluate the efficacy, safety, and health benefits of regular consumption of the composition in adults with Diabetes Mellitus Type II and any of the cohort conditions: Hypertension, dyslipidemia, gastritis, and constipation.

[0049] Clinical study is initiated and guidelines of the Indian Council of Medical Research (ICMR) India regulations with the said investigational product F1 (TT Compounded Asafoetida) product amongst 80 adults aged between 30 to 65 years, all sexes inclusive with history of uncomplicated Diabetes Mellitus type II with a minimum of one of the cohort conditions as Dyslipidemia, Gastritis, Constipation and Hypertension. As per the defined test protocol 1g of investigational formulation (F1) (TT Compound Asafoetida powder) to be consumed in a 300ml of the beverage, which is to be prepared as per the standardized instructions for a period of 96 days.

[0050] As per the research methodology 20 subjects acted as control group who consumed only beverage without the investigational product. Assessment of Fasting blood glucose, post prandial blood glucose, HbA1C, fasting triglycerides, Amlapitta Symptoms, Constipation Severity is done on periodic intervals with day 0, day 48 (interim) and day 96 (completion). The results of the study concluded with statistical findings that 91.23% of subjects showed who consumed the investigational formulation (F1) (TT Compounded Asafoetida Powder) showed 20% reduction in HbA1C in 96 days. The average reduction is 21.39% compared to 0.88% in Control Group. The results of the cohort conditions showed that 81.25% of subjects who consumed the investigational formulation (F1) (TT Compounded Asafoetida Powder) showed a statistically significant reduction in Gastritis as assessed by Amlapitta Symptoms Score in 96 days, compared to Control Group. Likewise, 92.85% of subjects who consumed the investigational formulation (F1) (TT Compounded Asafoetida Powder) showed a statistically significant reduction in Constipation as assessed by Constipation Severity Scale in 96 days, compared to Control Group.

[0051] The foregoing description is a specific embodiment of the present disclosure. It should be appreciated that this embodiment is described for purpose of illustration only, and that numerous alterations and modifications may be practiced by those skilled in the art without departing from the spirit and scope of the invention. It is intended that all such modifications and alterations be included insofar as they come within the scope of the invention as claimed or the equivalents thereof.
, Claims:We Claim:

1) A ferula-based formulation consisting of ferula extracts, wherein the ferula extracts comprises of is prepared from a variety of Ferula sp., including Ferula asafoetida, Ferula alliacea, Ferula rubricaulis, Ferula foetida, Ferula foetidissima, Ferula jaeschkeana, Ferula fukanensis, Ferula sinkiangensis and Ferula narthex, either alone or as a combination of two or more species.

2) The ferula-based formulation, as claimed in claim 1, comprising of edible constituents selected from a texturizer, bulking agent or thickener, colouring agent, chemical or natural preservative, other plant extracts, taste and flavor enhancer, antimicrobial agent, antioxidants, pH modulator, minerals, desiccant, sweetener, vitamins, carbonating agents or any combination thereof.

3) The ferula-based formulation, as claimed in claim 1, wherein the ferula extracts are made utilizing ferula resin or a component of the plant, such as leaves, rhizomes, shoots, stems, or roots, or any combination thereof.

4) The ferula-based formulation, as claimed in claim 3, wherein process of obtaining ferula resins entails cutting the root or rhizomes, scraping the exudates after a few days, and repeating the exudation process up to the first few determined cuts (incisions) on top of root or rhizomes.

5) The ferula-based formulation, as claimed in claim 3, wherein fresh, dried, or a combination of both forms of the ferula resin and/or plant component may be used.

6) The ferula-based formulation, as claimed in claim 3, wherein the ferula resin and/or plant material is used directly or after grinding.

7) The ferula-based formulation, as claimed in claim 3, wherein a solvent-based ferula resin extract and/or a solvent-based plant component extract is used separately or in combination.

8) The ferula-based formulation, as claimed in claim 3, wherein the extract of ferula is produced by combining weighed portions of the resin and/ or solvent -based extract of the resin or plant and/or ground plant material with a solvent of pH 4-9 at a solvent to resin ratio ranging from 0.5:1 to 30:1, while incubating at a temperature of 20-40 °C in presence of air for about 10-30 minutes.

9) The ferula-based formulation, as claimed in claim 1, wherein the formulation comprises of not more than 27 parts of F. asafoetida, 0.1-15 parts of F. foetida, 0.025 – 18 parts of F. alliacea, 0.3-16 parts of F. foetidissima, 0.02-23 parts of F. rubricaulis, 0.1-20 parts of F. narthex, 0.02-21 parts of F. jaeschkeana, 0.02-23 parts of F. fukanensis, 0.25-15 parts of F.sinkiangensis.

10) The ferula-based formulation, as claimed in claim 1, further comprising plant gums such as gum arabic or synthetic emulsifiers and stabilizers preferably containing polysaccharides and glycoproteins which act as prebiotics.

11) The ferula-based formulation, as claimed in claim 1, further comprises of one or more edible oil or fats, preferably mustard oil, sunflower, groundnut, safflower oil, sesame oil and the like that act as emulsifiers or a binding agent.

12) The ferula-based formulation, as claimed in claim 1, further comprises of one or more binding or bulking agents such as edible starch derived from cereal flour and not limited to wheat flour, refined wheat flour, soya flour, corn flour, and rice flour.

13) The ferula-based formulation, as claimed in claim 1, further comprises of carbohydrates of 60-80%, moisture of 10-20%, protein of 4-12%, fat of 0.5-2%, ash content of 1-15%, alcoholic extractable matter of 4-8%, and volatile oil of 0.2-5%.

14) The ferula-based formulation, as claimed in claim 13, wherein the volatile oil contains volatile compounds including 20- 60% of (E)-sec-Butyl propenyl disulfide, 1-35% of (Z)-sec-Butyl propenyl disulfide, 1-40% 1-(1-(Methylthio)propyl)-2-propyldisulfane, 0.1- 10% of Disulfide, bis(1-methylpropyl), 0.1- 10% of n-Propyl sec-butyl disulfide, 0.1- 9% alpha-Pinene, 0.1- 8% of beta-Ocimene, < 2% of a-Phellandrene, < 2% of ?-Terpinene, < 2% of p-Cymene, < 2% of D-Limonene and others.

15) The ferula-based formulation, as claimed in claim 1, further comprises phenolic acids including vanillin, ferulic acid, coumarins, as well as their esters, polyacetylenes, and other metabolites.

16) The ferula-based formulation, as claimed in claim 1, wherein the formulation is prepared in form of powder either compressed or loose, granules, lumps, capsules, cake and liquid or suspensions such as solutions, syrups, sauce, oils.

17) The Ferula-based formulation, as claimed in claim 1, when ingested alone or in combination with other preparations or drinks, may produce health advantages by regulating blood sugar levels, improving dyslipidemia and/or gastritis, and/or preventing constipation.

18) A method for preparation of ferula-based formulation, wherein the method comprises a homogeneous mixture of ingredients, and wherein the Ferula sp. resins and/or plant extracts are at a level of 0.1- 27% w/w with a particle size less than 500 µm, plant gums are at a level of 50- 75% w/w with a particle size less than 500 µm, edible oil at 0.1- 20% v/w, edible starch at a level of 30- 50% w/w with a particle size less than 500 µm, and wherein the homogenous mixture of ingredients is blended for 10-30 minutes, and aged for 12-72 hours in air at the temperature ranging between 22- 40°C.

19) A method for preparing flavored beverage from a ferula-based formulation, comprising steps of adding starter culture at a level of 0.5–5% to pasteurized milk that has been cooled to 40–50°C, incubating the mixture at 25–45°C for 4–8 hours, and churning coagulated product with a solvent (pH: 6–8) at a ratio of 1:1 until a homogeneous suspension of pH 3.5–5 is obtained, and to which the Ferula-based formulation is added at a level of 0.05-5% w/v.

20) A method for preparing flavored beverage from a ferula-based formulation, comprising steps of adding starter culture at a concentration of 0.5-5% to pasteurized milk that has been cooled to 35-50°C, followed by adding Ferula-based formulation at a concentration of 0.05-7% w/v and thoroughly mixing at a rpm of 50-200 for about 10-20 minutes and incubating the mixture at 25-45 °C for 4-8 hours before churning the product with an extracting medium (pH: 6-8) at a ratio of 1:1 until a homogeneous suspension of pH 3.5–5.3 is obtained.