The present disclosure relates generally to compositions for management of urolithiasis. More specifically, the disclosure is directed to a functional beverage composition for nephroprotective effect against urolithiasis comprising Dolichos biflorus, Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania somnifera, Foeniculum vulgare Mill, and stevia. The present disclosure further provides a process of preparing the composition.
BACKGROUND OF THE INVENTION
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. [0003] Urolithiasis afflicts around 19% of the Asian population with variable recurrence rate, 21 to 53%. Recurrence of renal stones treatments may eventually lead to deterioration of renal functional. Calcium oxalate (75%-90%) is the most common type of stone that afflicts Asian population and its proportion has increased in countries, like China, Japan, India and Indonesia. Oxalate is known as a metabolic end-product (largely produced by liver and less likely absorbed from food) handled by membrane oxalate transporters. Recent studies in Slc26 transporter knockout mice reported manifestation of hyperoxalemia, hyperoxaluria and calcium oxalate urolithiasis and suggested patho-physiological role of membrane oxalate transporters dysfunction. A study on 114 patients of idiopathic calcium oxalate urolithiasis suggested an inheritable malfunctioning in oxalate membrane transport in stone cases.
[0004] Pharmacological and interventional therapeutic procedures do not overcome the risk of stone recurrence and therapy like Extracorporeal Shock Wave Lithotripsy (ESWL) may also be associated with many serious adverse effects including hypertension, renal damage and impaired functioning. Few years back, the use of medicinal plants was proposed for better management of urolithiasis. Till date, plenty of research has been done in this discipline but could

not provide satisfactory results. All together few adverse events are reported with use of compounds extracted from medicinal plants. With the recent developments in nutrition science, it was suggested as "Let functional food be thy medicine" and thus, there is requirement to switch from the nutritional aspect of food to functional foods, as forms of dietary therapy to cure diseases (Martirosyan DM, Singh J. A new definition of functional food by FFC: what makes a new definition unique? Functional foods in health and disease. 2015 Jul l;5(6):209-23 and Olaiya CO, Soetan KO, Esan AM. The role of nutraceuticals, functional foods and value added food products in the prevention and treatment of chronic diseases. African Journal of food science. 2016 Oct 31;10(10): 185-93). [0005] Thus, there is a need in the art to develop new and improved composition for management of urolithiasis employing natural components with efficacy and no adverse side effects.
OBJECTS OF THE INVENTION
[0006] An object of the present disclosure is to provide a composition for
management of urolithiasis.
[0007] An object of the present disclosure is to provide a functional beverage
composition for management of urolithiasis comprising natural components.
[0008] An object of the present disclosure is to provide a functional beverage
composition that restores normalization of citrate and calcium and does not affect
plasma and urinary levels of creatinine, phosphorous and urea.
[0009] Another object of the present disclosure is to provide a process for
preparation of the composition.
SUMMARY OF THE INVENTION
[0010] This summary is provided to introduce a selection of concepts in a simplified form that are further described below in Detailed Description section. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.

[0011] Aspects of the present disclosure provide a composition comprising
natural plant based components that effectively manage the condition of
urolithiasis. The present disclosure provides a functional beverage composition
for the same.
[0012] In an aspect, the present disclosure provides a functional beverage
composition for management of urolithiasis comprising Dolichos biflorus,
Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania
somnifera, Foeniculum vulgare Mill, and stevia.
[0013] In an embodiment, the composition may comprise whole of Dolichos
biflorus, whole of Phyllanthus emblica L., whole of Ocimum tenuiflorum L, whole
of Withania somnifera and whole of Foeniculum vulgare Mill.
[0014] In an embodiment, the composition may comprise an extract of Dolichos
biflorus, extract of Phyllanthus emblica L., extract of Ocimum tenuiflorum L,
extract of Withania somnifera and extract of Foeniculum vulgare Mill.
[0015] In an embodiment, the composition may comprise part of Dolichos
biflorus, part of Phyllanthus emblica L., part of Ocimum tenuiflorum L, part of
Withania somnifera and part of Foeniculum vulgare Mill.
[0016] In an embodiment, the part or extract may be selected from the group
comprising of root, leaves, shoot, fruits, rhizome, seed, stem, barks, flower, sap,
bud or combinations thereof of the plant.
[0017] In an embodiment, the present disclosure provides a functional beverage
composition for management of urolithiasis comprising Dolichos biflorus seeds,
Phyllanthus emblica L. fruit, Ocimum tenuiflorum L. leaves, green tea leaves,
Withania somnifera roots, Foeniculum vulgare Mill, seeds, and stevia.
[0018] In an embodiment, the present disclosure provides a functional beverage
formulation for management of urolithiasis comprising Dolichos biflorus,
Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania
somnifera, Foeniculum vulgare Mill, and stevia.
[0019] In a preferred embodiment, the formulation is a tea precursor formulation.
[0020] In an aspect, the present disclosure provides a process of preparing a
functional beverage composition comprising the steps of: (a) powdering and

optionally sieving Dolichos biflorus seeds, dried Phyllanthus emblica L. fruit, dried Ocimum tenuiflorum L. leaves, dried green tea leaves, dried Withania somnifera roots, Foeniculum vulgare Mill, seeds, and dried stevia leaves to get uniform sized powders; and (b) mixing the powders of step (a) for about 30 minutes to give the composition.
[0021] Other aspects of the invention will be set forth in the description which follows, and in part will be apparent from the description, or may be learnt by the practice of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0022] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0023] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply. [0024] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0025] In some embodiments, numbers have been used for quantifying weights, percentages, ratios, and so forth, to describe and claim certain embodiments of the invention and are to be understood as being modified in some instances by the term "about." Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements. [0026] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0027] As used in the description herein and throughout the claims that follow, the meaning of "a," "an," and "the" includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of "in" includes "in" and "on" unless the context clearly dictates otherwise. [0028] Unless the context requires otherwise, throughout the specification which follow, the word "comprise" and variations thereof, such as, "comprises" and "comprising" are to be construed in an open, inclusive sense that is as "including, but not limited to."
[0029] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein.

[0030] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. "such as") provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention. [0031] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified.
[0032] The description that follows, and the embodiments described therein, is provided by way of illustration of an example, or examples, of particular embodiments of the principles and aspects of the present disclosure. These examples are provided for the purposes of explanation, and not of limitation, of those principles and of the disclosure.
[0033] It should also be appreciated that the present disclosure can be implemented in numerous ways, including as a system, a method or a device. In this specification, these implementations, or any other form that the invention may take, may be referred to as processes. In general, the order of the steps of the disclosed processes may be altered within the scope of the invention. [0034] The headings and abstract of the invention provided herein are for convenience only and do not interpret the scope or meaning of the embodiments. [0035] The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include all possible combinations of the disclosed elements. Thus, if one embodiment comprises elements A, B, and C, and a second embodiment

comprises elements B and D, then the inventive subject matter is also considered
to include other remaining combinations of A, B, C, or D, even if not explicitly
disclosed.
[0036] As described herein, the term 'therapeutically effective amount' refers to
the amount of the composition required to bring about a change or improvement
in a subject without side effects or overdosing.
[0037] The term, "subject" as used herein refers to an animal, preferably a
mammal, and most preferably a human. The term "mammal" used herein refers to
warm-blooded vertebrate animals of the class 'mammalia' , including humans,
characterized by a covering of hair on the skin and, in the female, milk-producing
mammary glands for nourishing the young, the term mammal includes animals
such as cat, dog, rabbit, bear, fox, wolf, monkey, deer, mouse, pig and human.
[0038] The term, 'management', or 'treatment' as used herein refers to alleviate,
slow the progression, attenuation, prophylaxis or as such treat the existing disease
or condition. Treatment also includes treating, preventing development of, or
alleviating to some extent, one or more of the symptoms of the diseases or
condition.
[0039] In an embodiment, the present disclosure provides a functional beverage
composition for management of urolithiasis comprising Dolichos biflorus,
Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania
somnifera, Foeniculum vulgare Mill, and stevia.
[0040] In an embodiment, the composition may comprise whole of Dolichos
biflorus, whole of Phyllanthus emblica L., whole of Ocimum tenuiflorum L, whole
of Withania somnifera and whole of Foeniculum vulgare Mill.
[0041] In an embodiment, the composition may comprise an extract of Dolichos
biflorus, extract of Phyllanthus emblica L., extract of Ocimum tenuiflorum L,
extract of Withania somnifera and extract of Foeniculum vulgare Mill.
[0042] In an embodiment, the composition may comprise part of Dolichos
biflorus, part of Phyllanthus emblica L., part of Ocimum tenuiflorum L, part of
Withania somnifera and part of Foeniculum vulgare Mill.

[0043] In an embodiment, the part or extract may be selected from the group
comprising of root, leaves, shoot, fruits, rhizome, seed, stem, barks, flower, sap,
bud or combinations thereof of the plant.
[0044] In a preferred embodiment, the composition comprises Dolichos biflorus
seeds, Phyllanthus emblica L. fruit, Ocimum tenuiflorum L. leaves, green tea
leaves, Withania somnifera roots, Foeniculum vulgare Mill, seeds, and stevia. In a
more preferred embodiment, the composition comprises Dolichos biflorus seeds,
Phyllanthus emblica L. dried fruit, Ocimum tenuiflorum L. dried leaves, green tea
dried leaves, Withania somnifera dried root powder, Foeniculum vulgare Mill.
Seed powder, and stevia dried leaves.
[0045] Dolichos biflorus seeds are a rich source of various natural bioactive
substances and have potential for treating variety of diseases. The inventors of the
present disclosure studied the medicinal properties of the seeds for providing a
suitable composition for effective management of urolithiasis.
[0046] In an embodiment, the present disclosure comprises Dolichos biflorus in a
weight percentage range of about 58% to about 62%, preferably a range of about
59%) to about 61%> of the composition.
[0047] In an embodiment, the present disclosure comprises Phyllanthus emblica
L. in a weight percentage range of about 9% to about 11%, preferably a range of
about 9.5%o to about 10.5% of the composition.
[0048] In an embodiment, the present disclosure comprises Ocimum tenuiflorum
L. in a weight percentage range of about 9% to about 11%, preferably a range of
about 9.5%o to about 10.5% of the composition.
[0049] In an embodiment, the present disclosure comprises green tea leaves in a
weight percentage range of about 7% to about 9%, preferably a range of about
7.5%o to about 8.5% of the composition.
[0050] In an embodiment, the present disclosure comprises Withania somnifera in
a weight percentage range of about 4% to about 6%, preferably a range of about
4.5%o to about 5.5% of the composition.

[0051] In an embodiment, the present disclosure comprises Foeniculum vulgare Mill, in a weight percentage range of about 4% to about 6%, preferably a range of about 4.5% to about 5.5% of the composition.
[0052] In an embodiment, the present disclosure comprises stevia in a weight percentage range of about 1.5% to about 2.5%, preferably a range of about 1.9% to about 2.1%) of the composition.
[0053] In an embodiment, the composition comprises Dolichos biflorus in a weight percentage range of about 58% to about 62%, Phyllanthus emblica L. in a weight percentage range of about 9% to about 11%, Ocimum tenuiflorum L in a weight percentage range of about 9% to about 11%, green tea leaves in a weight percentage range of about 7% to about 9%, Withania somnifera in a weight percentage range of about 4% to about 6%, Foeniculum vulgare Mill, in a weight percentage range of about 4% to about 6%, and stevia in a weight percentage range of about 1.5% to about 2.5%.
[0054] In an embodiment, the present disclosure provides a functional beverage formulation for management of urolithiasis comprising Dolichos biflorus, Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania somnifera, Foeniculum vulgare Mill, and stevia.
[0055] In an embodiment, the formulation may further comprise one or more excipient(s). The excipient may be selected from the group comprising of flavonoids, vitamins, stabilizers, binders, flavoring agents, sweeteners, coloring agents, humectants, preservatives, olfactory agents, and combinations thereof. Any other excipient(s) well known to a person skilled in the art may be employed without going beyond the spirit and scope of the disclosure. The choice and amount of the excipient to be employed may be determined based on the formulation being prepared.
[0056] In a preferred embodiment, the formulation is a tea precursor formulation. [0057] The tea precursor formulation may be infused in an infusion solvent to give tea. The infusion solvent may be milk, water, or combination thereof, preferably hot water, or any solvent suitable for drinking that does not interfere with the functioning of the phyto-chemicals. The tea gets infused in the solvent in

about 3 to 10 minutes. Said formulation may additionally be consumed with artificial sweeteners, honey, or sugar, preferably honey may be taken with the tea. [0058] The composition of the present disclosure restores normal blood and urinary oxalate and citrate levels and shows antiurolithiatic activity. It is believed that increase in plasma and urinary oxalate is primarily due to endogenous production instead of intestinal absorption of oxalate, in case of defective Slc26 transporters. Without being bound to theory, it is believed that endogenous oxalate in conjunction with idiopathic or chronic reduced activity or defective Slc26 transporters due to aging results in calcium oxalate kidney stones. Slc26 family transporters facilitate oxalate secretion from kidney and intestine and help to maintain oxalate homeostasis (Brzica H, Breljak D, Burckhardt BC, Burckhardt G, Sabolic I. Oxalate: from the environment to kidney stones. Arhiv za higijenuradaitoksikologiju. 2013 Dec 6;64(4):609-0). The composition of the present disclosure is believed to exhibit antiurolithiatic activity through improving activity of Slc26 transporters. Treatment with probenecid (Slc26 transporters inhibitor) blocks the antiurolithiatic effect exhibited by the composition and these findings indicate that the composition possibly produces antiurolithiatic effect through Slc26 transporters.
[0059] The composition significantly reduces blood and urine oxalate level and does not affect plasma and urinary level of creatinine, phosphorous and urea. Dolichos biflorus, the key active ingredient, exhibits antiurolithiatic effect through restoration of oxalate homeostasis by improving activity of Slc26 family transporters. This corroborates that Dolichos biflorus bioactive compounds possibly partly shift oxalate excretion from kidney to intestine through increasing activity of Slc26 family transporters and thereby reduce hyperoxaluria and hyperoxalemia.
[0060] In some embodiments, the composition possesses cytoprotective effect and antioxidant activity.
[0061] The composition has no adverse side-effects, is eco-friendly, economical and easy to produce. The composition has high patient compliance as it can be easily consumed as a beverage and is palatable.

[0062] In still another embodiment, the present disclosure relates to a tea bag
comprising the tea precursor formulation and a means for packaging.
[0063] In an embodiment, the means for packaging may be any suitable material
that is porous and is suitable for infusion. The means for packaging may be made
from cotton or muslin, cotton silk, jute, nylon, paper, cellulose, alloy or metal,
preferably it is a muslin cloth. The means for packaging must not interfere with
the aroma or flavor of the tea.
[0064] In an embodiment, the present disclosure provides a process of preparing a
functional beverage composition comprising the steps of: (a) powdering and
optionally sieving Dolichos biflorus seeds, dried Phyllanthus emblica L. fruit,
dried Ocimum tenuiflorum L. leaves, dried green tea leaves, dried Withania
somnifera roots, Foeniculum vulgare Mill, seeds, and dried stevia leaves to get
uniform sized powders; and (b) mixing the powders of step (a) for about 30
minutes to give the composition.
[0065] In an embodiment, the composition obtained by the above process may be
stored in a clean, dry place away from moisture and light. It may also be packaged
into tea bags for future use.
[0066] In an embodiment, the present disclosure provides a method of treatment,
prevention and amelioration of urolithiasis comprising administering a
therapeutically effective amount of the composition to a subject.
[0067] The composition is also effective in treatment of hyperoxalemia,
hyperoxaluria, and urolithiasis caused by calcium oxalate. Chronic mild
hyperoxaluria can cause calcium oxalate stone formation or urolithiasis.
[0068] While the foregoing describes various embodiments of the disclosure,
other and further embodiments of the disclosure may be devised without departing
from the basic scope thereof. The scope of the invention is determined by the
claims that follow. The invention is not limited to the described embodiments,
versions or examples, which are included to enable a person having ordinary skill
in the art to make and use the invention when combined with information and
knowledge available to the person having ordinary skill in the art.

EXAMPLES
[0069] The present invention is further explained in the form of following examples. However, it is to be understood that the following examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. [0070] MATERIALS AND METHODS: All the chemicals and solutions used for present study were of analytical grade purchased from Magus Chemicals, Mohali and reagents were prepared fresh before use. All plant constituents were collected from Dehradun, Uttarakhand during the month of June 2017. The voucher specimen of all constituents: Dolichos biflorus (seeds- voucher no. 0443), Phyllanthus emblica L. (dried fruit- voucher no. 0924), Ocimum tenuiflorum L. (dried leaves- voucher no. 0773), Green Tea: (dried leaves- voucher no. 0331), Withania somnifera (dried root powder- voucher no. 0578), Foeniculum vulgare Mill. (Seed's powder- voucher no. 0808) and Stevia (dried leaves- voucher no. 0511) was fixed into herbarium sheet, authenticated by registered plant taxonomist and deposited at the department of botany, Sri Venkateshwara University, Tirupati, Andhra Pradesh and College of Pharmacy, Chitkara University, Rajpura. The seeds were powdered and passed through a mesh 60 and stored in airtight container prior to extraction.
[0071] Statistical Analysis: Values were expressed in terms of mean ± standard error mean. Differences among data were determined using one- way ANOVA test followed by Tukey's multiple comparison test (GraphPad Software, Inc., version 6, CA, USA.) and P < 0.05 was considered statistically significant. Example 1: Preparation of functional beverage composition as Tea precursor formulation
[0072] Each 2 g tea bag constituted of: moderately coarse powder of Dolichos biflorus (seeds) (1.2 g), Phyllanthus emblica L. (dried fruit) (0.2 g), Ocimum tenuiflorum L. (dried leaves) (0.2 g), Green Tea: (dried leaves) (0.16 g), Withania somnifera (dried root powder) (0.1 g), Foeniculum vulgare Mill. (Seed's powder) (0.1 g) and Stevia (dried leaves) (0.04 g). The seeds and dried leaves were powdered and passed through a mesh 60 to ensure powder of uniform particle size and stored in airtight container prior to mixing. The uniformed particle size

powders were added in a stainless steel pan as per their composition. The powders were transferred to tumbler mixing machine and allowed to mix for 30 minutes to give the tea precursor formulation. The mixed powder was weighed and filled 2g in each tea bag. The filled tea bag was dipped in 250 mL hot water (temperature 90 °C) for 10 minutes. Tea bag was removed after 10 minutes and the aqueous extract was allowed to cool (at room temperature), filtered with filter paper to give the tea formulation, which was tested for antiurolithiatic activity. Example 2: Antiurolithiatic activity
[0073] 2.1 Animals: Antiurolithiasis activity of the functional beverage was evaluated in albino Wistar rats using ethylene glycol induced urolithiasis model. Male Wistar rats weighing around 200-250 g were obtained from animal house of College of Pharmacy, Chitkara University, Rajpura. The study protocol and all the experimental procedures used in this study were approved from the Institutional Animal Ethics Committee. The animals were kept in cages of polypropylene and maintained under standardized conditions (temperature 27°C ± 1°C, humidity 60% ± 4%>, and natural dark and light cycles), fed with diet and water as per protocol. The rats were divided into following groups, comprising of six animals in each group. Naive control served as the control regimen and received regular food and drinking water. Remaining groups received urolithiasis inducing treatment till 28th day, comprising ethylene glycol (1% w/v) with ammonium chloride (1% w/v) for initial 4 days, followed by ethylene glycol (1% w/v) alone in drinking water. Urolithiasis control group received calcium oxalate crystals- inducing treatment till 28th day. Vehicle group received vehicle only along with calcium oxalate crystals - inducing treatment till 28th day. Cystone group served as the standard regimen and received antiurolithiatic drug, cystone (500 mg/kg, po, OD), from 15th day till 28th day. Dolichos biflorus group received the tea formulation of Example 1 (2 ml po, TDD) from 15th day till 28th day and Thiazide group received hydrochlorothiazide diuretics (200 mg/kg, OD) from 15th day till 28th day. Probenecid groups (on regular diets and oxalate-free diets) received probenecid (100 mg/kg ip, OD) from 15th day till 28th day. In another group (on regular diets and oxalate-free diets), probenecid (100 mg/kg ip,

OD) was administered along with the tea formulation of Example 1 (2 ml po, TID) from 15th day till 28th day.
[0074] 2.2 Collection and Analysis of Urine; After 28 days of a treatment period, all rats were placed in individual metabolic cages with free access to drinking water, but without food. Urine samples were collected for 24 hours. The urine volume of each animal was measured. Urine sample of each animal was centrifuged at 2500g for 15 minutes. In the supernatant, total urinary excretion of oxalate, citrate, calcium, phosphorous, and creatinine was measured by various biochemical kits (Magus Chemicals, Mohali, India) according to manufacturer's guidance.
[0075] Table 1 presents the effect of the formulation and other treatment groups on urinary biochemical parameters.
Table 1: Effect on urinary biochemical parameters

Naive
contro
1 Urolit
hiasis
contro
1 Vehicl
e contro
1 Presen t
compo sition (FB)
Group Cyston
e treated Thiazi
de treate
d Probe
necid
treate
d Probe necid + FB Probe necid +oxala te free diet Probe necid + FB
+
oxalat
e free
diet
Oxalat
e
(uinol/
L) 238.2± 18.93 2881± 259.5a 3065± 129.5a 331.8± 43.09b 460.5± 53.63b 2489± 197.8 2407± 212.6 2880± 146.4 2511± 200.7 2566± 230.4
Citrat
e (uinol/
L) 4.53±0 .20 2.48±0 .32a 2.68±0
.33a 4.78±0. 28bl 4.21±0.
46b2 3.31± 0.26 3.43±0 .26 3.35±0 .27 3.03± 0.34 3.05±0
.25
Calciu
m (uinol/
L) 1.4±0. 14 0.10±0 .02al 0.10±0 .01al 1.61±0.
19b2 1.56±0.
23b3 1.00± 0.16b3 1.48±0 .19b3 1.15±0
.27 1.22± 0.35 1.50±0 .30
Magn esium (mmol
/L) 3.23±0 .22 6.35±0 .34a 5.96±0 .38a 3.13±0. 31b 3.30±0.
35b 2.68± 0.21b 6.21±0 .36 5.05±0 .39 5.41± 0.55 5.63±0 .40
Creati
nine
(mmol
/L) 4.35±0 .28 3.55±0 .55 3.71±0 .33 3.95±0. 31 3.96±0. 44 4.18± 0.34 4.21±0 .43 4.18±0 .41 4.30± 0.41 4.43±0 .27
Phosp
hate
(mmol
/L) 14.88± 0.99 16.35± 1.31 15.15± 1.33 14.88± 1.06 16.20± 0.90 17.08 ±1.60 16.43± 1.09 16.87± 1.80 15.77 ±1.32 17.30± 1.77
Volum
e (ml/24 hours) 29.27± 1.32 27.48± 1.97 23.50± 1.53 29.52± 1.52 25.57± 2.02 28.40 ±0.70 26.17± 2.04 25.32± 1.96 27.07 ±2.11 26.55± 1.51

ap<0.0001 vs Naive control; alp<0.01 vs Naive control; bp<0.0001 vs Urolithiasis control; blp<0.001 vs Urolithiasis control; b2p<0.01 vs Urolithiasis control; b3p<0.05 vs Urolithiasis control; Values are expressed as Mean±SEM; N=6
[0076] The present formulation and cystone significantly increased urinary level of citrate as compared to urolithiasis control, vehicle control and hydrochlorothiazide treated group. However, there was no statistically significant difference between present composition and cystone treated groups for urinary level of citrate. The probenecid treatment antagonized the beneficial effect of the formulation on urinary citrate level.
[0077] Present formulation and cystone significantly reduced urinary oxalate level as compared to urolithiasis control, vehicle control and hydrochlorothiazide treated group. However, there was no statistically significant difference between the present composition and cystone treated groups for urinary level of oxalate. The probenecid treatment antagonized the beneficial effect of the present formulation on urinary oxalate level.
[0078] The present formulation and cystone significantly increased urinary calcium level as compared to urolithiasis control and vehicle control group. However, there was no statistically significant difference between the formulation, cystone and hydrochlorothiazide treated groups for urinary level of calcium. However, probenecid treatment did not antagonize the beneficial effect of the present formulation on urinary calcium level.
[0079] Also, the administration of oxalate free diet with probenecid did not further reduce urinary and blood oxalate level in animals treated with or without the present formulation. This supports that endogenous oxalate instead of dietary oxalate contributes to primary source of oxalate in body. Therefore, low oxalate or oxalate free diet contributes less to urolithiaisis management. [0080] Functional Beverage (FB) and cystone did not significantly affect urinary creatinine, phosphorous and volume as compared to naive control, urolithiasis control, vehicle control and hydrochlorothiazide treated group. Therefore, the aqueous functional beverage of Example 1 was found to be equieffective to cystone, which is a clinically established treatment of urolithiasis.

[0081] 2.3 Serum Analysis: On 29th day, each rat under treatment was anesthetized with diethyl ether and blood sample was collected retro-orbitally and thereafter animals were sacrificed by cervical decapitation. Serum was separated by centrifugation at 10,000g for 5min and stored at -20 °C, until analysis. Various blood parameters oxalate and citrate including serum parameters such as phosporous, calcium, urea, and creatinine were analyzed by using various biochemical kits (Magus Chemicals, Mohali, India) according to the manufacturer's guidance.
[0082] Table 2 below presents the effect of the different treatment groups on the plasma biochemical parameters.
Table 2: Effect on plasma biochemical parameters

Vehicle Control Urolith
iasis
Contro
1 Cystone Treated Thiazi
de Treate
d Functio
nal
beverag
e(FB)
composi
tion Proben
ecid
Treate
d Proben ecid +
FB
Treate
d Proben
ecid
+oxalat
e free
diet
Treate
d Proben
ecid +
FB +
Oxalat
e free
diet
Treate
d
Oxalate
(junol/L
) 16.16±1 .70 61.83±
3.52 18.00±3 .05a 59.50± 5.17 16.33±1 .54a 72.83± 5.10b 63.66± 6.81b 56.33± 5.23b 46.83± 5.62b
Citrate
(junol/L
) 133.00±
3.72 78.33± 3.34 125.16± 6.88a 87.00± 5.30 130.50±
5.25a 76.83± 4.79 83.83± 5.13b 78.33± 4.30b 86.16± 6.74b
Calcium (mg/dL) 9.16±0. 94 10.33± 0.88 9.50±0. 67 11.50± 1.43 9.50±0. 88 10.00± 0.57 11.08± 0.66 9.86±0.
77 9.98±0.
77
Creatini
ne (mg/dL) 0.46±0. 08 1.98±0. 21 0.45±0. 09 0.35±0 .07 0.55±0. 10 1.10±0.
23 1.33±0. 21 1.31±0. 20 1.25±0. 20
Phospho
rous (mg/dL) 8.83±0. 70 6.96±0. 82 7.83±0. 94 8.00±0 .89 8.16±0. 60 7.50±0. 99 8.38±1. 04 8.56±1. 06 9.15±1. 04
Urea (mg/dL) 13.66±1 .11 13.01± 1.22 11.66±1 .11 10.66± 1.02 10.33±0 .49 12.66± 0.88 13.43± 0.86 13.08± 1.36 12.98± 1.05
ap<0.0001 vs Urolithiasis control; bp<0.0001 vs Functional Beverage Group; Values are expressed as Mean±SD; N=6
[0083] Aqueous functional beverage (FB) formulation and cystone significantly increased citrate level in blood as compared to urolithiasis control, vehicle control

and hydrochlorothiazide treated group. However, there was no statistically significant difference between the present composition and cystone treated groups for citrate level in blood.
[0084] The FB formulation and cystone significantly reduced oxalate level in blood as compared to urolithiasis control, vehicle control and hydrochlorothiazide treated group. However, there was no statistically significant difference between FB formulation and cystone treated groups for oxalate level in blood. [0085] The inhibitor of Slc26a6 transporter, probenecid treatment antagonized the effect of the present formulation on restoration of normal level of citrate and oxalate in blood. The oxalate free diet along with probenecid with and without FB could not restore normal level of citrate and oxalate in blood. These findings indicated dietary oxalate content did not significantly contribute in hyperoxaluria and/or hyperoxalemia. The impairment in oxalate excretion through Slc26a6 transporter may be the possible reason for increased urinary oxalate level followed by urolithiasis. Thus, treatment with the present composition restores functioning of Slc26a6 transporter at intestinal and renal system and thereby restores normal level of oxalate in blood.
[0086] The present composition and cystone did not significantly affect blood calcium, creatinine, phosphorous and urea as compared to naive control, urolithiasis control, vehicle control and hydrochlorothiazide treated group. [0087] 2.4 Histopathological Study of Kidney: The abdomen of the animal was cut and both kidneys were removed. Kidneys after isolation, were cleaned with phosphate buffer saline (ice-cold). Kidney of animals was dissected and fixed in 10% neutral buffered formalin and processed in a sequential manner with graded alcohol and xylene, and in the end embedded in paraffin wax. Histological sections of about 5 /mi thickness were prepared by microtomy and stained with hematoxylineosin (H&E) dye for histological examination. Histological slides were analyzed under a light microscope at lOx magnification. The various histological changes such as hemorrhages, congestion, tubular swelling, vacuolar changes in the cytoplasm, and interstitial fibrosis were observed to check changes in the kidney.

[0088] Histopathological sections under light microscope showed normal architecture in vehicle control rats. In urolithiatic, thiazide, probenecid and probenecid + oxalate free diet animals, mild nephritis with renal tubular damage and inflammation was observed due to concomitant treatment of ethylene glycol in drinking water. Cystone, present formulation (FB) and Probenecid + FB treatment with and without oxalate free diet treated group, showed less mild inflammation and nephritis and reported protection against ethylene glycol induced nephrotoxicity. It showed that the present formulation and cystone possess cytoprotective effect which may be due to their antioxidant activity. Whereas thiazide and probenecid did not possess cytoprotective effect as these do not possess any antioxidant activity.
[0089] The foregoing examples are merely illustrative and are not to be taken as limitations upon the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the scope of the invention.
ADVANTAGES OF THE PRESENT INVENTION
[0090] The present disclosure provides a functional beverage composition for management of urolithiasis comprising natural components. [0091] The present disclosure provides a functional beverage composition that restores normalization of citrate and calcium and does not affect plasma and urinary levels of creatinine, phosphorous and urea.

We Claim:

1. A functional beverage composition for management of urolithiasis comprising Dolichos biflorus, Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania somnifera, Foeniculum vulgare Mill, and stevia.
2. The composition as claimed in claim 1, wherein the composition comprises whole of Dolichos biflorus, whole of Phyllanthus emblica L., whole of Ocimum tenuiflorum L, whole of Withania somnifera and whole of Foeniculum vulgare Mill.
3. The composition as claimed in claim 1, wherein the composition comprises an extract of Dolichos biflorus, extract of Phyllanthus emblica L., extract of Ocimum tenuiflorum L, extract of Withania somnifera and extract of Foeniculum vulgare Mill.
4. The composition as claimed in claim 1, wherein the composition comprises part of Dolichos biflorus, part of Phyllanthus emblica L., part of Ocimum tenuiflorum L, part of Withania somnifera and part of Foeniculum vulgare Mill.

5. The composition as claimed in claims 3-4, wherein the part or extract is selected from the group comprising of root, leaves, shoot, fruits, rhizome, seed, stem, barks, flower, sap, bud or combinations thereof of the plant.
6. The composition as claimed in claim 1, wherein the composition comprises Dolichos biflorus seeds, Phyllanthus emblica L. fruit, Ocimum tenuiflorum L. leaves, green tea leaves, Withania somnifera roots, Foeniculum vulgare Mill. seeds, and stevia.
7. The composition as claimed in claim 1, wherein the composition comprises Dolichos biflorus in a weight percentage range of 58% to 62%, Phyllanthus emblica L. in a weight percentage range of 9% to 11%, Ocimum tenuiflorum L in a weight percentage range of 9% to 11%, green tea leaves in a weight percentage range of 7% to 9%, Withania somnifera in a weight percentage range of 4% to 6%, Foeniculum vulgare Mill, in a weight percentage range of 4% to 6%, and stevia in a weight percentage range of 1.5% to 2.5%.

8. A functional beverage formulation for management of urolithiasis comprising Dolichos biflorus, Phyllanthus emblica L., Ocimum tenuiflorum L, green tea leaves, Withania somnifera, Foeniculum vulgare Mill, and stevia.
9. The formulation as claimed in claim 8, wherein the formulation is a tea precursor formulation.
10. A process of preparing a functional beverage composition comprising the
steps of: (a) powdering and optionally sieving Dolichos biflorus seeds, dried
Phyllanthus emblica L. fruit, dried Ocimum tenuiflorum L. leaves, dried green tea
leaves, dried Withania somnifera roots, Foeniculum vulgare Mill, seeds, and dried
stevia leaves to get uniform sized powders; and (b) mixing the powders of step (a)
for 30 minutes to give the composition.