Description:TECHNICAL FIELD
[0001] The present invention relates to the field of herbal drug formulations, nutraceuticals, and functional foods. More particularly, it relates to an aqueous extract of Hibiscus sabdariffa L. exhibiting potent antioxidant, anthelmintic, and antidiabetic activities, and to the formulation of an herbal jelly incorporating the extract for human consumption.
BACKGROUND ART
[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] Herbal plants have long been recognized for their medicinal properties in traditional systems of medicine. Among them, Hibiscus sabdariffa L. (family: Malvaceae) is widely used for its anthocyanins, flavonoids, and phenolic compounds. The calyces are consumed in teas, beverages, and traditional remedies. Herbal medicines and nutraceuticals have gained significant attention as alternatives to synthetic drugs due to their safety, accessibility, and lower side effects. Among the wide variety of medicinal plants, Hibiscus sabdariffa L., commonly known as “Roselle,” is traditionally used in many cultures as a herbal remedy for hypertension, inflammation, and metabolic disorders. Its calyces are rich in anthocyanins, flavonoids, phenolic acids, and organic acids, which have been reported to exhibit antioxidant, antimicrobial, antidiabetic, and anthelmintic activities.
[0004] Conventional synthetic formulations for antioxidant and antidiabetic therapy often cause undesirable effects and are expensive for regular use, particularly in developing regions. While Hibiscus sabdariffa has been studied for its pharmacological potential, there is a lack of standardized aqueous extraction processes and integrated formulations that combine its bioactivity with convenient consumer-friendly dosage forms.
[0005] However, conventional applications are limited to crude usage, with little evidence-based formulation development or standardized evaluation. The prior art discloses certain antioxidant and antidiabetic properties of hibiscus extracts, but systematic aqueous extraction, phytochemical profiling (GC-MS), and integrated evaluation of antioxidant, anthelmintic, and antidiabetic activity are lacking. Furthermore, incorporation of such extracts into a palatable, stable jelly formulation with demonstrated efficacy and consumer acceptance is not reported in the existing literature.
[0006] CN102813686B The invention discloses a method for extracting active ingredients in roselle calyx, which belongs to the technical field of extracting active ingredients in roselle. The extraction process is as follows: dry roselle calyx is crushed, put into a reaction kettle liner of a subcritical water extraction tank, and press The liquid-solid ratio is 15:1~25:1, add water with a pH value of 1.0~3.0, the extraction pressure is 5~10MPa, the extraction temperature is 110~150°C, the extraction time is 20~50min, filter, and the obtained filtrate is concentrated to dryness under reduced pressure. After adding water to dissolve, freeze-dry to obtain the active ingredient extract of roselle calyx. The method has the advantages that: compared with the traditional water extraction method and 30% alcohol extraction method, the method for extracting effective components in the roselle calyx by using subcritical water has shorter extraction time, higher extract yield, and is green and pollution-free.
[0007] Hibiscus sabdariffa L. – A phytochemical and pharmacological review. In an aspect of the prior art, Hibiscus sabdariffa (roselle) has been extensively studied for its phytochemical composition, particularly anthocyanins, phenolic acids and various volatile/non-volatile constituents. Comprehensive reviews and phytochemical surveys report that anthocyanins (e.g., delphinidin and cyanidin glycosides) are major pigments in the calyces and that non-anthocyanin compounds such as 5-hydroxymethylfurfural (HMF) and various furfural derivatives are commonly detected by chromatographic profiling. This body of analytical work establishes the typical chemical makeup of roselle calyces and provides common analytical markers used for extract standardization.
[0008] Existing methods of herbal product preparation also face challenges such as poor reproducibility, lack of stability, and limited patient compliance. Herbal teas or decoctions made from Hibiscus sabdariffa are popular, but they are not standardized, and their bioactive content may vary depending on preparation. Moreover, functional food products incorporating Hibiscus sabdariffa are limited in variety and often lack combined health benefits targeting multiple biological pathways.
[0009] Thus, there exists a need for a standardized aqueous extract of Hibiscus sabdariffa with proven multifunctional bioactivities and its application in the development of an herbal jelly suitable for nutraceutical and therapeutic use.
[0010] The present invention provides an aqueous extract of Hibiscus sabdariffa L. calyces exhibiting potent antioxidant, anthelmintic, and antidiabetic activities. The extract is obtained by refluxing dried calyces’ powder with distilled water, yielding a phytochemically rich fraction containing anthocyanins, furans, organic acids, flavonoids, and glycosides. GC-MS profiling confirms multiple bioactive compounds.
OBJECTS OF THE INVENTION
[0011] The principal object of the present invention is to overcome the disadvantages of the prior art.
[0012] The primary objective of the present invention is to provide a standardized aqueous extract of Hibiscus sabdariffa L. calyces exhibiting multifunctional bioactivities, specifically antioxidant, anthelmintic, and antidiabetic properties, and to develop a novel herbal jelly formulation incorporating the extract for use as a nutraceutical functional food.
[0013] Another objective of the present invention is to perform phytochemical screening and GC-MS analysis of the extract to identify bioactive compounds.
[0014] Another objective of the present invention is to establish in vitro evaluation methods including DPPH assay for antioxidant activity, earthworm model for anthelmintic activity, and yeast cell glucose uptake for antidiabetic activity.
[0015] Another objective of the present invention is to formulate a novel herbal jelly incorporating Hibiscus sabdariffa extract in combination with natural ingredients such as sappan wood, chia seeds, and pineapple.
[0016] Another objective of the present invention is to provide a consumer-acceptable nutraceutical formulation with demonstrated health-promoting effects.
SUMMARY
[0017] The present invention provides a novel process for the aqueous extraction of Hibiscus sabdariffa calyces and the subsequent evaluation of its antioxidant, anthelmintic, and antidiabetic properties. The invention further discloses the formulation of a multi-ingredient herbal jelly incorporating Hibiscus sabdariffa extract along with Caesalpinia sappan (sappan wood), chia seeds, corn flour, and pineapple juice.
[0018] In one aspect, the method involves collecting authenticated Hibiscus sabdariffa calyces, preparing dried powder, and subjecting the powder to Soxhlet extraction using distilled water. The pooled aqueous extracts are concentrated, screened for phytochemical constituents, and analyzed by GC-MS to identify bioactive compounds. The extract is further assessed for antioxidant activity using the DPPH method, anthelmintic activity against selected parasites, and antidiabetic potential via glucose uptake by yeast cells.
[0019] In another aspect, the invention discloses a herbal jelly formulation, wherein standardized Hibiscus sabdariffa extract is combined with aqueous extracts of sappan wood, chia seeds, pineapple juice, and corn flour slurry. The ingredients are blended under controlled heating with continuous stirring until a homogenous jelly texture is obtained. The jelly is molded, cooled, refrigerated, and stored in airtight containers for subsequent use.
[0020] The present invention provides a cost-effective, natural, and multifunctional nutraceutical product with proven biological activities, offering potential applications in healthcare, dietary supplementation, and functional food industries.
[0021] The invention provides a herbal jelly formulation comprising hibiscus extract, sappan wood extract, chia seeds, pineapple juice, corn flour, and sugar, which retains the bioactivity and offers high palatability. The jelly formulation received overwhelmingly positive consumer feedback, supporting its use as a nutraceutical functional food.
BRIEF DESCRIPTION OF DRAWINGS
[0022] The accompanying illustrations are incorporated into and form a part of this specification in order to aid in comprehending the current disclosure. The pictures demonstrate exemplary implementations of the current disclosure and, along with the description, assist to clarify its fundamental ideas.
[0023] Fig.1 (a) (b) (c) (d) (e) shows the extraction process of Hibiscus sabdariffa calyces.
[0024] Fig 2 (a) (b) shows the GC-MS chromatogram of aqueous extract showing major bioactive compounds.
[0025] Fig 3 shows the DPPH antioxidant activity curve.
[0026] Fig 4 shows the Anthelmintic activity comparison between extract and Albendazole.
[0027] Fig 5 shows the Glucose uptake curve of yeast cells treated with extract.
[0028] Fig 6 shows the process diagram of herbal jelly formulation.
[0029] Fig 7 shows the Pie chart showing consumer feedback on jelly.
[0030] It should be noted that the figures are not drawn to scale, and the elements of similar structure and functions are generally represented by like reference numerals for illustrative purposes throughout the figures. It should be noted that the figures do not illustrate every aspect of the described embodiment sand do not limit the scope of the present disclosure.
[0031] Other objects, advantages, and novel features of the invention will become apparent from the following detailed description of the present embodiment when taken in conjunction with the accompanying drawings.
DETAILED DESCRIPTION OF THE INVENTION
[0032] 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.
[0033] As used throughout this description, the word "may" is 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 documents 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.
[0034] 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.
[0035] The present invention is described hereinafter by various embodiments with reference to the accompanying drawing, wherein reference numerals used in the accompanying drawing correspond to the like elements throughout the description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiment set forth herein. Rather, the embodiment is provided so that this disclosure will be thorough and complete and will fully convey the scope of the invention to those skilled in the art. In the following detailed description, numeric values and ranges are provided for various aspects of the implementations described. These values and ranges are to be treated as examples only and are not intended to limit the scope of the claims. In addition, several materials are identified as suitable for various facets of the implementations.
[0036] The present invention relates to the field of herbal drug formulations, nutraceuticals, and functional foods. More particularly, it relates to an aqueous extract of Hibiscus sabdariffa L. exhibiting potent antioxidant, anthelmintic, and antidiabetic activities, and to the formulation of an herbal jelly incorporating the extract for human consumption.
[0037] In an embodiment of the present invention a formulation of a herbal jelly includes aqueous extract of Hibiscus sabdariffa, comprising, i) 6 g of sappan wood extract prepared by boiling in water and reducing to half its volume; ii) 1.6 g of Hibiscus sabdariffa extract prepared by boiling in water and filtering; iii) 1.6 g of chia seeds, pre-soaked in water; iv) 46 g of corn flour dissolved in 150 mL of water to form slurry; v) 80 g of pineapple pulp sweetened with 8 g sugar and filtered into juice; and water as required, characterized in that the extracts, chia seeds, corn flour slurry, and pineapple juice are combined by heating and continuous stirring until the mixture thickens to jelly consistency, and the resulting jelly is cooled, molded, refrigerated, and stored in airtight containers.
[0038] In another embodiment of the present invention the method for preparing an aqueous extract of Hibiscus sabdariffa calyces and investigating its in vitro antioxidant, anthelmintic, and antidiabetic properties,
comprising the steps of: collecting fresh and dried powder of Hibiscus sabdariffa calyces; storing said powder in a clean, airtight container; subjecting the dried powder to Soxhlet extraction using distilled water as the aqueous solvent, wherein 25 g of powder is refluxed with 150 mL of water for 4 hours, followed by further extraction with 250 mL of water for 3 hours, and drying the residue prior to subsequent cycles; and pooling and concentrating the extracts by rotary evaporation; characterized in that the aqueous extract is screened for phytochemical constituents, analyzed by GC-MS for bioactive compounds, and evaluated in vitro for, antioxidant activity using the DPPH radical scavenging assay; anthelmintic activity against selected organisms; and antidiabetic potential through glucose uptake by yeast cells.
[0039] In an embodiment of the present invention, microscopic evaluation of the powdered form of Hibiscus sabdariffa calyces was carried out to identify diagnostic features and confirm authenticity of the raw material. A small quantity of the powdered sample was placed in a watch glass and mixed with chloral hydrate solution. The mixture was gently heated to achieve clearing of tissues. Following the clearing process, a few drops of phloroglucinol solution were added, immediately followed by concentrated hydrochloric acid, and the preparation was subjected to mild heating to selectively stain lignified cells. Thereafter, a small portion of the stained material was transferred onto a clean glass slide, covered with a cover slip, and observed under a compound microscope. The microscopic observation revealed characteristic anatomical and histological features suitable for confirming the identity and quality of the plant material.
Feature Description
1.Epidermal cells Polygonal or rectangular shaped cells with slightly wavy walls, often visible in surface fragments.
2.Trichomes Multicellular, uniseriate trichomes often seen with 1–2 cells long; some may be glandular or non-glandular.
3. Calcium oxalate crystals Present as prismatic crystals or scattered druses; often seen singly or in groups.
4. Red colouring matter Due to anthocyanins (mainly delphinidin and cyanidin glycosides); gives pinkish-red tone to powder. Visible as stained regions.
5. Parenchymatous cells Thin-walled, isodiametric or rounded cells containing red pigment; may also contain starch grains.
6. Vessels (Xylem Elements) Long, narrow, with spiral or scalariform thickening; visible as elongated structures.
7. Fibres Elongated, lignified structures with tapering end, often slightly curved; add mechanical strength.
8.Starch grains Simple or compound, round to oval in shape; though not always prominent in calyces.
9. Glandular Structures Occasionally visible as capitate glands; stalked with multicellular heads.
10. Pigmented cells Cells with deep red to purple pigment (anthocyanin); may be seen scattered or aggregated.
[0001] In an embodiment of the present invention, extraction of the powdered calyces of Hibiscus sabdariffa was carried out using an aqueous solvent system. Specifically, 25 grams of dried and powdered Hibiscus sabdariffa calyces were subjected to reflux extraction with 150 mL of distilled water for 4 hours. The marc obtained after the first extraction was air-dried and subsequently refluxed again with 250 mL of distilled water for an additional 3 hours. The extracts obtained from both reflux cycles were pooled together to ensure maximum yield of phytoconstituents. The combined extract was then concentrated under reduced pressure using a rotary evaporator to remove excess solvent, yielding a thick, semi-solid aqueous extract.
[0002] The dried extract was subjected to preliminary phytochemical screening to identify the major classes of bioactive compounds. The percentage yield of the extract was calculated with respect to the weight of the initial dried powder material, and the results are presented in the accompanying table.
Extraction Yield obtained Percentage yield %
4 hrs 5.5g 20%
3 hrs 9.5g 38%
Total 15g 60%
[0003] In an embodiment of the present invention, the extract of Hibiscus sabdariffa was subjected to carbohydrate detection tests. In Molisch’s test, the filtrate treated with alcoholic α-naphthol solution followed by the careful addition of concentrated sulphuric acid produced a violet ring at the junction, thereby confirming the presence of carbohydrates. In Benedict’s test, the filtrate treated with Benedict’s reagent and heated in a water bath resulted in the formation of an orange-red precipitate, indicating the presence of reducing sugars. In Barfoed’s test, the filtrate treated with Barfoed’s reagent and boiled for 1–2 minutes yielded a dark red precipitate, thereby confirming the presence of carbohydrates. In Seliwanoff’s test, the filtrate treated with Seliwanoff’s reagent and subjected to heating developed a cherry-red coloration, confirming the presence of ketose sugars.
[0004] In another aspect of the invention, the extract was tested for alkaloids using Wagner’s reagent. The treatment of the extract with Wagner’s reagent produced a reddish-brown precipitate, confirming the presence of alkaloids. Similarly, in a further embodiment, the extract was evaluated for proteins using the Biuret test. The addition of sodium hydroxide solution and copper sulfate solution to the extract, followed by gentle mixing and standing for 4–5 minutes, led to the appearance of a bluish-violet color, thereby confirming the presence of proteins.
[0005] In yet another embodiment of the present invention, the extract was subjected to tannin detection by means of the ferric chloride test. The addition of ferric chloride solution produced a blue, black, violet, or green coloration/precipitate, indicating the presence of tannins. In addition, the extract was examined for flavonoids using ferric chloride, and the formation of a green to black coloration confirmed the presence of flavonoids.
[0006] In a further aspect of the invention, the extract was tested for glycosides using the Keller-Killiani method. The extract treated with water and glacial acetic acid, followed by ferric chloride, and subsequently layered with concentrated sulphuric acid along the sides of the test tube, produced a brown ring at the junction of the liquid layers. This characteristic reaction confirmed the presence of glycosides.
[0007] GC-MS Peak Report: Compound Name and Area
S. No. Compound Name Area
1 Furfural 9137845
2 2(5H)-Furanone 16580518
3 6-Oxa-bicyclo[3.1.0]hexan-3-one 1686292
4 2,5-Furandione, dihydro-3-methylene- 1894834
5 2-Amino-1-(furan-2-yl)ethan-1-ol, Me derivative 3384494
6 2-Furancarboxaldehyde, 5-methyl- 4756471
7 2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3-one 748167
8 2-Methyliminoperhydro-1,3-oxazine 1516264
9 Propanoic acid 29598199
10 Succinic anhydride 5679543
11 2,3-Pentanedione, 4-methyl- 963208
12 5H-1,4-Dioxepin, 2,3-dihydro-2,5-dimethyl- 696217
13 Pentanoic acid, 4-oxo- 1569436
14 2,3-Dihydro-1H-pyrazol-3-one, 2Ac derivative (isomer 2) 2962966
15 2-Furancarboxylic acid, 2-methylpropyl ester 4232437
16 4-Methyloctanoic acid 1789963
17 2,5-Furandione, 3-methyl- 2246695
18 1,6-Diazabicyclo(3.1.0)hexane-5-carboxylic acid-, methyl ester 9197323
19 sec-Butyl acetate 922915
20 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl- 6851055
21 5-(Methoxymethyl)furan-2-carbaldehyde 1419452
22 Benzoic acid 1097107
23 2-(Isobutoxymethyl)oxirane 4059438
24 2-Furancarboxaldehyde, 5-(chloromethyl)- 1140108
25 3-Cyclopentene-1-acetaldehyde, 2-oxo- 928992
26 2-Pentenoic acid, 3-ethyl-, methyl ester 3874521
27 6-Hydroxy-2,6-dihydropyran-3-one 1938576
28 1H-Pyrazole-3-carboxylic acid, 2,5-dihydro-5-oxo- 5598123
29 5-Hydroxymethylfurfural 60893864
30 Cycloheptane-1,2-dione 1073072
31 5-(Hydroxymethyl)-2-(dimethoxymethyl)furan 2371380
32 3-Heptanol 900710
33 .beta.-D-Glucopyranose, 1,6-anhydro- 9405024
34 1,6-Anhydro-.beta.-D-glucofuranose 3993418
35 Davanone B 3657389
36 (-)-Spathulenol 3390618
37 Globulol 977460
38 Bicyclo[2.2.1]heptane-2,3-diol, 1,7,7-trimethyl-, (endo,endo)- 1724228
39 Ledol 688368
40 beta.-Eudesmo 954314
41 Davanone-2-ol, beta- 3340860
42 2-Undecanone, 6,10-dimethyl- 1589006
43 1-Hexadecanol 1087754
44 Hexadecanoic acid, methyl ester 763368
45 1-Eicosanol 1769951
46 9-Octadecenoic acid (Z)-, methyl ester 3674371
47 Glycidyl palmitate 2018267
48 Glycidyl palmitoleate 14481075
49 Acetic acid, 3-methyl-6-oxo-9-oxabicyclo[3.3.1]non-2-yl ester 986366
50 Glycidyl palmitate 1203154
[0008] In an embodiment of the present invention, the anthelmintic activity of the aqueous extract of Hibiscus sabdariffa calyces was evaluated using an in-vitro assay model. The test was conducted employing Indian adult earthworms (Lumbricus terrestris), which are widely accepted as experimental models owing to their anatomical and physiological resemblance to human intestinal parasites, particularly for preliminary anthelmintic screening.
[0009] In an aspect of the invention, the earthworms were collected from the agricultural fields of SRMC Agricultural College and maintained under suitable laboratory conditions prior to experimentation. The assay was performed by preparing test solutions of the aqueous extract at concentrations of 20 mg/mL and 40 mg/mL in distilled water. Groups of five earthworms of approximately equal size were introduced into separate Petri dishes containing 25 mL of the respective test solutions.
[0010] In another embodiment of the present invention, albendazole at a concentration of 40 mg/mL was employed as the reference standard drug, while double-distilled water was used as the control. All test and standard solutions were prepared freshly immediately prior to the initiation of the experiment in order to maintain reproducibility and accuracy.
[0011] In a further embodiment, the evaluation parameters included the time required for paralysis and the time required for death of the worms after exposure to the test solutions. These values were recorded for each group and compared against the standard drug and control. The results demonstrated the anthelmintic potential of Hibiscus sabdariffa extract relative to albendazole, thereby confirming its therapeutic relevance.
Sample group Concentration Test organism Earthworm
Time for Paralysis
(min) Time for death(min)
Contol (Distilled water) - - -
Standard(Albendazole) 40mg/ml 3.25 7.45
Hibiscus Extract 20mg/ml 3.03 6.02
Hibiscus Extract 40mg/ml 2.1 3.25
[0012] In an embodiment of the present invention, the anthelmintic potential of the aqueous extract of Hibiscus sabdariffa calyces was evaluated using an in-vitro earthworm assay. Indian adult earthworms (Lumbricus terrestris) were selected due to their physiological similarity to human intestinal parasites. Groups of five worms of approximately equal size were exposed to test solutions of the extract prepared at concentrations of 20 mg/mL and 40 mg/mL in distilled water. Albendazole at 40 mg/mL served as the reference standard, while double-distilled water was used as a control.
[0013] In an aspect of the invention, the time required for paralysis and death of worms was recorded for each group. The results demonstrated that Hibiscus sabdariffa extract exhibited a dose-dependent paralytic and lethal effect on the worms. Higher concentrations induced faster paralysis and death compared to lower concentrations. Notably, the extract at both 20 mg/mL and 40 mg/mL acted more rapidly than albendazole at 40 mg/mL, indicating superior activity.
[0014] In another embodiment, dose–response analysis indicated an IC₅₀ value of approximately 0.05 mg/mL, confirming high potency of the extract. The dose–response curve of Hibiscus sabdariffa extract versus albendazole for worm mortality further substantiated the significant anthelmintic effect. Increasing the concentration of the extract led to a marked reduction in time to death and an enhanced percent effect, thereby confirming concentration-dependent efficacy.
[0015] In a further embodiment of the present invention, the antioxidant activity of the extract was assessed using the in-vitro DPPH radical scavenging assay. The method was based on the reduction of the stable, purple-colored radical 1,1-diphenyl-2-picryl hydrazyl (DPPH) to a yellow-colored product in the presence of an antioxidant.
[0016] In an aspect of the invention, the reagents used were as follows: (i) DPPH solution prepared in methanol to an absorbance of 0.9 at 516 nm; (ii) ascorbic acid solution, prepared as a standard antioxidant (50 mg in 100 mL methanol); and (iii) the aqueous extract of Hibiscus sabdariffa, prepared at a concentration of 1 mg/mL.
Concentration(mg/ml) Mean absorbance Activity in percentage
1
0.07
93%
1
1
2
0.17
82.4%
2
2
3
0.27
74.4%
3
3
4
0.39
60.4%
4
4
5
0.45
54.6%
5
5
[0017] The procedure comprised the addition of varying concentrations of test solution to 75 μL of DPPH in methanol, with the final volume adjusted to 3 mL. DPPH solution alone served as the blank. The mixtures were incubated for 15 minutes at room temperature, and absorbance was measured spectrophotometrically. The percentage inhibition was calculated relative to the blank.
[0018] The results showed that the extract exhibited concentration-dependent free radical scavenging activity, with maximum inhibition of 93% observed at the lowest tested concentration (1 mg/mL). The antioxidant effect gradually decreased at higher concentrations, with 82.4% at 2 mg/mL, 74.4% at 3 mg/mL, 60.4% at 4 mg/mL, and 54.6% at 5 mg/mL.
[0019] In another embodiment of the invention, the results indicated that the aqueous extract of Hibiscus sabdariffa possesses significant radical-scavenging potential, particularly at lower concentrations, consistent with the properties of antioxidant-rich phytoconstituents. Such activity demonstrates potential therapeutic and nutraceutical applications in mitigating oxidative stress.
[0020] In an embodiment of the present invention, the glucose adsorption capacity of the extract was evaluated using the yeast cell glucose uptake method, which serves as a simple in vitro model for screening antidiabetic activity. The principle of this method is based on the ability of yeast cells (Saccharomyces cerevisiae) to absorb glucose from the medium, where an increase in glucose uptake in the presence of the extract indicates potential antidiabetic activity, functioning in a manner similar to insulin-like action.
[0021] In an aspect of the invention, the experimental procedure involved the preparation of a yeast suspension by dissolving dry yeast in warm distilled water at 37°C, incubating for 10 minutes, and centrifuging at 3000 rpm for 5 minutes. The pellet was washed several times and resuspended to obtain a 10% yeast suspension. A 5 mM glucose solution was prepared, and reaction mixtures were set up containing 1 mL glucose solution, the test extract at concentrations ranging from 1–5 mg/mL, and 1 mL yeast suspension. Controls were prepared using distilled water instead of the extract. All mixtures were incubated at 37°C for 60 minutes, followed by estimation of glucose levels in the supernatant using colorimetric methods at 520 nm.
[0022] In another aspect of the invention, the results demonstrated that the extract showed strong glucose adsorption activity across all tested concentrations. The adsorption activity was highest at 2 mg/mL with 96.77% activity, while at 5 mg/mL it remained significant at 82.79%. The trend suggested effective binding of glucose molecules, which could help regulate postprandial blood glucose levels.
[0023] In an aspect of the invention, the conclusion of this evaluation confirmed that the tested extract exhibited high glucose adsorption capacity, thereby showing potential as a hypoglycemic agent. This property may be attributed to the presence of dietary fibers or bioactive compounds capable of binding glucose molecules, indicating the therapeutic potential of the extract in blood sugar management.
[0024] Herbal Jelly Formulation
[0025] In an embodiment of the present invention, a novel herbal jelly formulation was developed using Sappan wood, Hibiscus sabdariffa, Chia seeds, Corn flour, and Pineapple, with the objective of creating a palatable nutraceutical product enriched with antioxidant, anthelmintic, and hypoglycemic properties.
[0026] In an aspect of the invention, the preparation process involved sequential extraction of the plant materials. Sappan wood extract was obtained by boiling 6 grams of wood pieces in 120 mL water for 10 minutes and reducing the volume to 60 mL, followed by filtration. Hibiscus sabdariffa extract was prepared by boiling 1.6 grams of powder in 100 mL of water, reducing to 50 mL, adding 25 mL water, and boiling further before filtration. Pineapple juice was prepared by grinding 2 slices of pineapple with 8 grams of sugar, followed by filtration. Corn flour slurry was prepared by dissolving 46 grams of corn flour in 150 mL water.
[0027] In another aspect of the invention, the extracts were combined in a pan along with pre-soaked chia seeds, and the corn flour slurry was gradually added with continuous stirring. The mixture was cooked until thickened, after which pineapple juice was incorporated. The final jelly was poured into greased molds, cooled at room temperature, and refrigerated for 1 hour to set. The product was then cut into desired shapes and stored in airtight containers.
[0028] In an aspect of the invention, this formulation provides a functional jelly with therapeutic benefits, integrating the pharmacological activities of the herbal ingredients with an appealing edible form.
[0029] Feedback and Responses of Jelly
[0030] In an embodiment of the present invention, the formulated herbal jelly was subjected to sensory evaluation and consumer feedback. The responses were analyzed using graphical representations such as pie charts.
[0031] In an aspect of the invention, the feedback revealed that a majority of the respondents rated the jelly as Excellent, reflecting a high degree of consumer acceptance and satisfaction. A negligible percentage of respondents rated the jelly as Poor, while no respondents marked it as Good or Average.
[0032] In another aspect of the invention, these results indicate that the jelly formulation not only possesses therapeutic potential but also offers sensory appeal and market acceptability as a functional food product.
[0033] Further, the operations need not be performed in the disclosed order, although in some examples, an order may be preferred. Also, not all functions need to be performed to achieve the desired advantages of the disclosed system and method, and therefore not all functions are required.
[0034] 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 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:I/We Claim:
1. A formulation of a herbal jelly comprising aqueous extract of Hibiscus sabdariffa, comprising:
i) 6 g of sappan wood extract prepared by boiling in water and reducing to half its volume;
ii) 1.6 g of Hibiscus sabdariffa extract prepared by boiling in water and filtering;
iii) 1.6 g of chia seeds, pre-soaked in water;
iv) 46 g of corn flour dissolved in 150 mL of water to form slurry;
v) 80 g of pineapple pulp sweetened with 8 g sugar and filtered into juice; and water as required,
characterized in that
the extracts, chia seeds, corn flour slurry, and pineapple juice are combined by heating and continuous stirring until the mixture thickens to jelly consistency, and the resulting jelly is cooled, molded, refrigerated, and stored in airtight containers.
2. The method for preparing an aqueous extract of Hibiscus sabdariffa calyces as claimed in claim 1, comprising the steps of:
i) collecting fresh and dried powder of Hibiscus sabdariffa calyces;
ii) storing said powder in a clean, airtight container;
iii) subjecting the dried powder to soxhlet extraction using distilled water as the aqueous solvent, wherein 25 g of powder is refluxed with 150 mL of water for 4 hours, followed by further extraction with 250 mL of water for 3 hours, and drying the residue prior to subsequent cycles; and

iv) pooling and concentrating the extracts by rotary evaporation.
3. The method as claimed in claim 2, wherein the mixture is cooked for 10–15 minutes until uniform thickening occurs, ensuring homogeneous texture.
4. The method as claimed in claim 2, wherein a light layer of edible oil is applied to the mold to prevent sticking during cooling.
5. The composition as claimed in claim 1, wherein the final product is refrigerated for at least one hour prior to storage to achieve set consistency.