Description:TECHNICAL FIELD

[0001] The present invention relates to a radish leaf Powder, More especially Formulation and Evaluation of a Nutraceutical Tablet using Radish leaf powder by Wet Granulation Method.

BACKGROUND ART

[0002] Due to the change in eating habits, lifestyle, food consumption and production shows impact on health and increasing incidence of lifestyle diseases are the leading cause for Nutraceuticals consumption and the market is booming, nutraceuticals have a role in nutritional, immunological and physiological functions.
[0003] Nutraceuticals are products, which other than nutrition are also used as medicine. A nutraceutical product may be defined as a substance, which has physiological benefit or provides protection against chronic disease. Nutraceuticals may be used to improve health, delay the aging process, prevent chronic diseases, increase life expectancy, or support the structure or function of the body. Nowadays, nutraceuticals have received considerable interest due to potential nutritional, safety and therapeutic effects.
[0004] Raphanus sativus (Radish) is an annual herb of family Cruciferae or Brassicaceae, and consumed as vegetable, commonly known as Mooli. Almost all parts of the plant including leaves, seeds and roots are utilized in medicine.
[0005] Omar M Noman, et al, 2021, have reported that Raphanus sativus.L leaves extract shows inhibitory effect on cancer cell proliferation. The 70% ethanol extract followed by maceration procedure of Raphanus sativus L. leaves effect on the proliferation and viability of two breast (MCF-7 & MDA-MB-231), liver (Hep G2), and lung (A549) cancer cells were evaluated using MTT assay. From the above MTT assay results revealed that Raphanus sativus.L exhibited antiproliferative activity against all the tested cancer cells. The viability of all the treated cells decreased in dose dependent manner and Raphanus sativus.L leaf extract Posses highest antiproliferative efficiency against all cell lines with IC50 values of 217-453 microgram per millilitre.
[0006] Da-Hee Chung, et.al, 2012, studied the effect of ethyl acetate extracts of radish leaves on hypertension in 11week old spontaneously hypertensive rats (SHRs) Six wister rats were used as normotensive controls. The systolic blood pressure (SBP) of the SHRs showed a decreasing trend with the consumption of the radish leaf extract. The SBP of the group fed 90mg extract/kg body weight reduced from 214 mmHg to 166 mmHg and was significantly lower than that of the normotensive and hypertensive controls. The extract increased the concentration of NO in serum and the activities of antioxidant enzymes such as glutathione peroxidase and catalyse in red blood cells. It was observed that consumption of an Ethyl acetate extract of radish leaves may have antihypertensive effects in SHRs since it increases the serum concentration of NO and fecal concentration of Na+ and enhance antioxidant activity
OBJECTS OF THE INVENTION
[0007] The principal object of the present invention is to overcome the disadvantages of the prior art by providing
[0008] The main object of the present invention is to formulate and evaluate nutraceutical tablet using Radish leaf powder treatment and prevention, anti-aging qualities, and cancer prevention.
[0009] Another object of the present invention is to provide a formulation that aids in increasing income of farmers by enabling them to increase production of radish leaves.
[0010] Yet another object of the present invention is to provide a formulation which is rich in Vitamin C & K, Iron, Phosphorus and other phytochemicals which are helpful in preventing the onset of different diseases and their complications.
[0011] The foregoing and other objects of the present invention will become readily apparent upon further review of the following detailed description of the embodiments as illustrated in the accompanying drawings.
SUMMARY OF THE INVENTION

[0012] The present invention relates to a radish leaf Powder, More especially Formulation and Evaluation of a Nutraceutical Tablet using Radish leaf powder by Wet Granulation Method.

[0013] In an embodiment of the present invention a radish leaf water extract-based nutraceutical formulation, comprising, Radish Leaf Extract; Starch Paste; Magnesium Stearate; Lactose; Dry Starch powder; and Talc.

[0014] In another aspect of the present invention is in the form of a tablet. The method of preparation of formulation, comprising the following steps: i) Collecting fresh white radish leaves and shade draying said leaves on a cotton cloth for 7 days followed by drying in hot air oven; ii) Powdering and sieving said dried leaves using blender and sieve no 22; iii) Adding Desired quantities of Starch, Lactose, Magnesium stearate, Talc was weighed. These powders are passed through 60 mesh sieves; iv) Mixing the sifted powders and Radish leaf powder thoroughly in mortar with pestle until uniform blend is obtained.

[0015] While the invention has been described and shown with reference to the preferred embodiment, it will be apparent that variations might be possible that would fall within the scope of the present invention.
BRIEF DESCRIPTION OF DRAWINGS

[0016] So that the manner in which the above-recited features of the present invention can be understood in detail, a more particular description of the invention, briefly summarized above, may have been referred to by embodiments, some of which are illustrated in the appended drawings. It is to be noted, however, that the appended drawings illustrate only typical embodiments of this invention and are therefore not to be considered limiting of its scope, for the invention may admit to other equally effective embodiments.
[0017] These and other features, benefits, and advantages of the present invention will become apparent by reference to the following text figure, with like reference numbers referring to like structures across the views, wherein:
[0018] Figure 1 illustrates a Radish leaves before drying.
[0019] Figure 2 illustrates a Radish leaves after drying.
[0020] Figure 3 illustrates a Radish Nutraceutical tablet.
[0021] Figure 4 illustrates a ?max of Raphanus sativus.L leaf powder.
[0022] Figure 5 illustrates Calibration Curve for radish leaf powder at 264nm.
[0023] Figure 6 illustrates Percentage of drug release from the Nutraceutical Tablet.
[0024] Figure 7 illustrates a UV Spectra for Dissolution of Nutraceutical Tablet.

Figure 5 represents graphical representation of Dissolution studies of Nutraceutical tablets.
DETAILED DESCRIPTION OF THE INVENTION

[0025] While the present invention is described herein by way of example using embodiments and illustrative drawings, those skilled in the art will recognize that the invention is not limited to the embodiments of drawing or drawings described and are not intended to represent the scale of the various components. Further, some components that may form a part of the invention may not be illustrated in certain figures, for ease of illustration, and such omissions do not limit the embodiments outlined in any way. It should be understood that the drawings and the detailed description thereto are not intended to limit the invention to the particular form disclosed, but on the contrary, the invention is to cover all modifications, equivalents, and alternatives falling within the scope of the present invention as defined by the appended claim.
[0026] As used throughout this description, the word "may" be used in a permissive sense (i.e. meaning having the potential to), rather than the mandatory sense, (i.e. meaning must). Further, the words "a" or "an" mean "at least one” and the word “plurality” means “one or more” unless otherwise mentioned. Furthermore, the terminology and phraseology used herein are solely used for descriptive purposes and should not be construed as limiting in scope. Language such as "including," "comprising," "having," "containing," or "involving," and variations thereof, is intended to be broad and encompass the subject matter listed thereafter, equivalents, and additional subject matter not recited, and is not intended to exclude other additives, components, integers, or steps. Likewise, the term "comprising" is considered synonymous with the terms "including" or "containing" for applicable legal purposes. Any discussion of document acts, materials, devices, articles, and the like are included in the specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention.
[0027] In this disclosure, whenever a composition or an element or a group of elements is preceded with the transitional phrase “comprising”, it is understood that we also contemplate the same composition, element, or group of elements with transitional phrases “consisting of”, “consisting”, “selected from the group of consisting of, “including”, or “is” preceding the recitation of the composition, element or group of elements and vice versa.
[0028] The present invention relates to a radish leaf water extract-based nutraceutical formulation and method of preparation thereof for prevention and treatment of many diseases in a cost effective manner, so that people can afford easily and as little to no side effects.

[0029] In an embodiment of the present invention a radish leaf water extract-based nutraceutical formulation, comprising, Radish Leaf Extract in the range of 45 to 50%; Starch Paste in the range of 8.5 to 8.75%; Magnesium Stearate in the range of 1.5 to 2.5%; Lactose in the range of 20% to 21.5%; Dry Starch powder 5% to 6.25%; and Talc in the range of 10% to 11.25%.

[0030] In another aspect of the present invention is in the form of a tablet. The method of preparation of formulation, comprising the following steps: i) Collecting fresh white radish leaves and shade draying said leaves on a cotton cloth for 7 days followed by drying in hot air oven; ii) Powdering and sieving said dried leaves using blender and sieve no 22; iii) Adding Desired quantities of Starch, Lactose, Magnesium stearate, Talc was weighed. These powders are passed through 60 mesh sieves; iv) Mixing the sifted powders and Radish leaf powder thoroughly in mortar with pestle until uniform blend is obtained.

[0031] In an aspect of the present invention the exact composition of the formulation is presented in the table as mentioned below:
Sl. NO INGREDIENTS QUANTITY FOR ONE
TABLET (mg) QUANTITY FOR 80
TABLETS (gm)
1 Radish Leaf Powder 200 16
2 Starch Paste (10%) 35 2.8
3 Magnesium Stearate 10 0.8
4 Lactose 85 6.8
5 Dry Starch powder 25 2.0
6 Talc 45 3.6

[0032] A method for preparation of said formulation comprising the following steps: the present invention, the method for preparation of said formulation comprising the following steps: i) Collection of leaves; (ii) Preparation of Granulating Medium (10% Starch Paste); (iii) Sifting of Powder; (iv) Blending of Powders; (v) Wet Granulating (Screening); (vi) Drying; (vii) Dry Screening; (vii) Dry Blending; (viii) Punching; (ix) Punching.
[0033] In another aspect of the present invention the residue was used as active ingredient for nutraceutical tablet formulation. All other ingredients such as Radish Leaf Powder; Starch Paste (10%); Magnesium Stearate; Lactose; Dry Starch powder; Talc.
[0034] Leaves of white radish were collected, weigh about 3.5 kg and washed with water and shade dried for 7 days on the cotton cloth. After shade drying process, leaves weighed about 220gms and dried in hot air oven at 600c for 1hour, dried leaves were powdered using blender. Then powder was passed through sieve no 22 and finally 164gms of powder was obtained.
[0035] Nutraceutical tablets were prepared by wet granulation method. 10% Starch paste was used as granulating medium, Magnesium stearate and talc as Glidant and Lubricant, Dry starch powder & Sod. Starch Glycolate as Disintegrating agent, Lactose as Diluents, Micro Crystalline Cellulose & Poly Vinyl Pyrrolidine as binder.
[0036] Table discloses the percentage composition of nutraceutical tablet:

[0037] The method for preparation of said formulation comprising Starch; Radish leaf powder; Magnesium Stearate; Lactose; Dry Starch powder; Talc.
[0038] RESULTS
Phyto-chemical evaluation.
1. Detection of Alkaloids: -
a. Dragendorff’s test: Took 2ml of aqueous extract in a clean test tube, add Dragendorff’s reagent. Orange brown color precipitate indicates presence of Alkaloids.
b. Hager’s test: Took 2ml of aqueous extract in a clean test tube, add Hager’s reagent. Yellow color precipitate indicates the presence of Alkaloids.
c. Wagner’s test: Took 2ml of aqueous extract in a clean tube, add Wagner’s reagent. Red color precipitate indicates the presence of Alkaloids.
d. Mayer’s test: Took 2ml of aqueous extract in a clean tube, add Mayer’s reagent. No cream colour precipitate indicates the absence of Alkaloids.
2. Test for Steroids:- Took 2ml of aqueous extract in a clean test tube, add few drops of chloroform and equal amount of conc. sulfuric acid. Appearance of blue colour indicates the presence of Steroids.
3. Tannins and phenolic compounds:-
[0039] a. Took 2ml of aqueous extract in a clean test tubes, add Ferric chloride. Appearance of bluish black colour indicates the presence of Tannins and Phenolic compounds
[0040] b. Took 2ml of aqueous extract in a clean test tube, add lead acetate. No appearance of bluish yellow precipitate indicates the absence of Tannins and Phenolic
[0041] c. Took 2ml of aqueous extract in a clean test tube, add potassium chromate solution. Appearance of orange yellow precipitate indicates the presence of Tannins and Phenolic compounds.
[0042] Flavonoids:-Took 2ml of aqueous extract & mix with 5% Fecl3 (Ferric chloride) solution. Green or purple precipitate indicates the presence of Flavonoids.
[0043] Protein and aminoacids :-
[0044] a. Millon’s test: Took 2ml of aqueous extract in a clean test tube, add Millon’s reagent. Appearance of yellowish to red precipitate indicates the presence of Proteins and Amino acids.
[0045] b. Xanthoprotein test: Took 2ml of aqueous extract in a clean test tube, add Ethanol and conc. Nitric acid. Appearance of yellow colour indicates the presence of Proteins and Aminoacids.
[0046] Carbohydrates test:-a. Molisch’s Test: Took 2ml of aqueous extract in a clean test tube, add Molisch’s reagent. Appearance of violet colour ring at the junction of the test tube Indicates the presence of Carbohydrates.
[0047] b. Fehling’s Test: Took 2ml of aqueous extract in a clean test tube, add Fehling’s reagent. Appearance of green colour indicates the presence of Carbohydrates.
[0048] Glycoside test:- a. Borntrager’s test: Took 2ml of aqueous extract in a clean test tube and dissolve in 1ml of Benzene and add dil. Ammonia to the sample add Hydrochloric acid heat and filter. Rose pink to red colour is not obtained indicates the absence of Glycosides. b. Keller Killiani test: Took 2ml of aqueous extract in a clean test tube and treated with Glacial acetic acid shake well, add a trace amount of Ferric chloride solution and Conc. Sulfuric acid, Presence of blue colour in acetic acid layer and presence of red colour at junction of 2 liquids indicates the presence of Glycosides. c. Legal’s test: Took 2ml of aqueous extract in a clean test tube add water make it alkaline add few drops of Sod. nitroprusside, no blue colour indicates the absence of Glycosides.
[0049] Preformulation Studies of powdered blend:-
a. Angle of repose. Angle of repose was determined by using funnel method. The weighed blend was taken in a funnel and height was adjusted in such a way that tip of the funnel almost touches the apex of the heap. Blend was allowed to flow through the funnel and height of the heap of blend was measured. A circle around the heap was drawn to measure the radius of the cone. The angle of repose was calculated using the equation.
tan ? = h/r
Where, h=height of the pile (cm).
r=radius of the pile base (cm).
b. Rate of flow
[0050] Weighed quantity of blend was taken in a clean and dry funnel placing a cotton plug at neck of funnel. The cotton was removed and time required for the blend to flow out of the funnel was recorded.
c. Loose bulk density
[0051] Loose bulk density was determined by placing weighed quantity of blend into graduated measuring cylinder. Volume and weight were measured.
LBD = Weight of the powder/volume of the packing.
d. Tapped bulk density
[0052] The measuring cylinder contain weighed amount of blend was allowed to fall under its own weight on to a hard surface from the height of 5cm at two second intervals. The tapping was continued until no further change in volume was noted.
TBD = weight of the powder/tapped volume.
e. Compressibility Index
[0053] It was determined using following formula. Compressibility Index (%) = (TBD- LBD) x 100 / TBD.
f. Hausner ratio
[0054] It was determined by using formula. Hausner ratio = TBD/LBD.

Table 03: Relationship between flow and angle of repose , Carr’s index, Hausner ratio.
Flow Angle of repose Carr’s index(%) Hausner ratio
Excellent <25 5-15 1.00-1.11
Good 25-30 12-16 1.12-1.18
Fair to passable 30-40 18-21 1.19-1.34
Poor >40 23-35 1.35-1.45
Very poor 33-38 1.46-1.59
Extremely poor >40 >1.60

Evaluation of nutraceutical tablets (Post compression)
a. Colour and Appearance
[0055] The compressed tablets were examined for their colour and appearance.
b. Weight variation test
[0056] Weight variation was determined by weighing 20 tablet individually. Average weight was calculated from the total weight of all tablets. The individual weights are compared with the average weight then percentage deviation was calculated using the formula,
Percentage deviation = individual weight – average weight x 100 / average weight.
Table No :08
The percentage difference in the weight variation should be within the permissible limit.
Average weight (mg) Percentage deviation allowed (%)
80 or less 10.0
More than 80 and less than 250 7.5
250 or more 5.0

c. Hardness test
[0057] The hardness of the tablet was tested using Monsanto hardness tester. The tablet was placed across the diameter in between the spindle and the anvil. The knob was adjusted to hold the tablet in position. The pressure was increased slowly to break the tablet.
d. Friability test
[0058] It was tested by using Roche friabilator (4 minute at 25 rpm). 10 tablets were weighed collectively and placed in the chamber of friabilator after 100 rotations (4 minute), the tablets are taken out from the friabilator and intact tablets are again weighed collectively. Percentage friability was determined.
Percentage Friability = W1 –W2/ W1 ×100
e. Thickness and Diameter
[0059] Thickness and Diameter of the tablets were evaluated by using Vernier Calipers.
f. Disintegration test
[0060] The Disintegration test apparatus consist of a basket rack assembly containing six open ended glass tubes, A tablet was placed in each of six tubes in the basket. They are held vertically and bottom of the tube was covered with 10mesh screen. By the use of the motor the basket raises and lowers in the immersion fluid at a frequency of 28-32 cycles per minute. The wire screen was always maintained below the level of the fluid. The fluid temperature was maintained at 37±2°C throughout the test. The disintegration time was noted when all the tablets pass through the sieve. This time should comply with the time stated in the monograph for that tablet.
[0061] Disintegration for uncoated tablets is 15mins.
g. Dissolution test
[0062] In vitro testing of drug release from tablet is common step of quality control as well as an initial phase of the product development process. In vitro dissolution testing of oral dosage forms measures the dissolution rate of drug substance going from the solid state into solution per unit time under standardized conditions of temperature, pH, and composition.
1. Preparation of standard sub-stock solution.
[0063] 50mg of radish leaf powder was weighed accurately and dissolved it in 50ml of volumetric flask. In that above solution withdrawn 5ml of solution dissolved in 50ml volumetric flask. Concentration of sub stock solution is 100µg/ml.
2. Preparation of standard calibration curve.
[0064] From the above standard sub-stock solution 1, 2, 4, 6, 8, 10ml were withdrawn and transferred to 10ml volumetric flask and make up the volume up to 10ml to get a concentration of 10, 20, 40, 60, 80, 100 µg/ml respectively. The absorbance of each solution was measured by UV-Visible spectrophotometer at 264nm. The curve of concentration (x-axis) v/s absorbance (y-axis) was plotted.
In-vitro drug release.
[0065] a. Dissolution studies on nutraceutical tablet of radish leaves powder were conducted in USP Apparatus 2 i.e., Paddle Apparatus (Electro lab TDT-08L). The dissolution medium was distilled water 900ml. The paddle rotation speed was kept at 50 rpm for 60 minutes. Samples were analyzed by UV-Visible Spectrophotometer at 264nm. (Shimadzu UV-1800). Percentage of drug released from tablet was calculated.
[0066] b. The Nutraceutical tablet was placed in dissolution basket containing 900ml of distilled water temperature is maintained at 37±0.50c, paddle is rotated at 50rpm.
[0067] c. 5ml of samples are withdrawn at different time period of 2.5min, 5min, 7.5min, 10min. Samples are filtered on whatman filter paper, samples are analyzed at 264nm using UV-Visible Spectrophotometer.
[0068] Table No- 09:- Observed Phytochemical screening reports of Raphanus sativus.L leaf powder.
SL.
No. Phytoconstituents Raphanus sativus.L leaf powder.
1 Alkaloids +ve = Present
2 Steroids +ve = Present
3 Tannins and Phenolic compounds +ve = Present
4 Flavonoids +ve = Present
5 Proteins and Aminoacids +ve = Present
6 Carbohydrates +ve = Present
7 Glycosides +ve = Present

Table no. 10. Observed Preformulation studies of the powdered blend.

SL. No. Preformulation tests Results
1 Angle of repose 34.550
2 Rate of flow 1.030 gm/sec
3 Loose bulk density 0.496 gm/cc
4 Tapped bulk density 0.554 gm/cc
5 Compressibility index 10.469%
6 Hausner Ratio 1.116

Table No-11 :- Observed evaluation of Nutraceutical tablet
SL. No. Evaluation test Results
1 Colour and appearance Light green and Round shape.
2 Percent weight variation 1.51%
3 Hardness test 4.2 kg/cm2
4 Friability test 0.503%
5 Thickness 0.4 mm
6 Diameter 1.43 mm
7 Disintegration Time 6 minutes

3. In-vitro drug release.
In pharmaceutical industry, drug dissolution testing is routinely used to provide critical in vitro drug release information for both quality control purposes, to assess batch-to-batch consistency of tablets.
1. Calibration curve.
Table 12:- Reading of different concentration of radish leaves powder at 265nm.
Sl No Absorbance Concentration(ug)
1 0.023 10
2 0.049 20
3 0.081 40
4 0.138 60
5 0.148 80
6 0.198 100

Percentage of Drug release :-
[0069] The average percentage of drug release within 10mins as determined by the proposed Spectrophotometric method after in vitro dissolution of tablets is depicted in figure 15.
[0070] Dissolution studies of Nutraceutical tablets were preformed and the percentage of drug release was found to be 63.75% within 7.5 mins.
EVALUATION OF ANTHELMINTIC ACTIVITY OF NUTRACEUTICAL TABLET:-
[0071] The findings of the Nutraceutical radish leaf tablet anthelmintic activity test showed that it had various levels of anthelmintic activity. At all of the tested concentrations, the Nutraceutical tablet demonstrated worm death in addition to worm paralysis.
[0072] When compared to the conventional medicine piperazine citrate, which has a concentration of 10mg/ml, the tablet was shown to have the maximum activity at 30mg/ml (paralysis time) and 50mg/ml (death time).
[0073] Table no-13 :- Observed Anthelmintic activity of Nutraceutical tablet.
Sl.
No Group mg/ml Eisenia fetida earthworm.
Time taken for paralysis in min Time taken for death in min
1 Control
(double distilled water)
-
-
-
2 Standard 10 21 44
3 Nutraceutical tablet 10 57 95
20 55 113
30 42 123
40 54 127
50 45 110
60 50 111

[0074] Various modifications to these embodiments are apparent to those skilled in the art from the description and the accompanying drawings. The principles associated with the various embodiments described herein may be applied to other embodiments. Therefore, the description is not intended to be limited to the 5 embodiments shown along with the accompanying drawings but is to be providing the broadest scope consistent with the principles and the novel and inventive features disclosed or suggested herein. Accordingly, the invention is anticipated to hold on to all other such alternatives, modifications, and variations that fall within the scope of the present invention and appended claims.
, Claims:1) A radish leaf water extract based nutraceutical formulation, comprising;
i) Radish Leaf Extract in the range of 45 to 50%;
ii) Starch Paste in the range of 8.5 to 8.75%;
iii) Magnesium Stearate in the range of 1.5 to 2.5%;
iv) Lactose in the range of 20% to 21.5%;
v) Dry Starch powder 5% to 6.25%; and
vi) Talc in the range of 10% to 11.25%.
2) The nutraceutical formulation as claimed in claim 1, wherein said formulation is in the form of a tablet.
3) The method of preparation of formulation as claimed in claim 1, comprising the following steps:
i) Collecting fresh white radish leaves and shade draying said leaves on a cotton cloth for 7 days followed by drying in hot air oven;
ii) Powdering and sieving said dried leaves using blender and sieve no 22;
iii) Adding Desired quantities of Starch, Lactose, Magnesium stearate, Talc were weighed. These powders are passed through 60 mesh sieve;
iv) Mixing the sifted powders and Radish leaf powder thoroughly in mortar with pestle until uniform blend is obtained.
4) The method of preparation as claimed in claim 3, wherein said method involves dry blending and punching to get tablets of sufficient hardness.
5) The nutraceutical formulation as claimed in claim 1, the dissolution method for analyzing the nutraceutical tablet formulated from radish leaf powder was carried out.