The universe has supplied a vast supply of cures for human illnesses. Because they contain elements with therapeutic properties, medicinal plants have been utilized for ages as cures for many illnesses. 80% of people worldwide still primarily receive their medical treatment from conventional practitioners, according to the WHO. Currently, there is a growing interest in herbal medicines around the world, along with a rise in laboratory research into the pharmacological characteristics of bioactive ingredients and their potential to cure different ailments.(1) Gorakhmundi, often referred to as Sphaeranthus indicus Linn, is a fragrant, spreading herb that is a member of the Asteraceae family. It is widespread in India, growing in humid environments from the Kumaon highlands to Sikkim in the Himalayas, at heights of up to 1500 meters. This plant goes by several names, such as Mahashravani, Tapodhana, Munditika, Mundi, Shravana, Bhikshu, and Shravanashirshaka.(2) In the Indian system of medicine, the plant as a whole plant or its different parts like leaf, stem, bark, root, flower and seed are widely used for curing many diseases. The plant has been reported with different types of secondary metabolites such as eudesmanolides, sesquiterpenoids, sesquiterpene lactones, sesquiterpene acids, flavone glycosides, flavonoid C-glycosides, isoflavone glycoside, sterols, sterol glycoside, alkaloid, peptide alkaloids, amino acids and sugars. Essential oil has been isolated from flowers and whole plant. (3) The whole plant, its isolated secondary metabolites and different parts have been reported for ovicidal, antifeedant, anthelmintic, antimicrobial, antiviral, macrofilaricidal, larvicidal, analgesic, antipyretic, hepatoprotective, antitussive, wound healing, bronchodilatory, mast cell stabilizing activity, anxiolytic, neuroleptic, immunomodulatory, anti diabetic, antihyperlipidemic and antioxidant, antioxidant, central nervous system  anti arthritic, nephroprotective, anticonvulsant activities and many other activities. It is also effective for psoriasis.(4) A range of secondary metabolites are present in different regions of Sphaeranthus indicus, such as Eudesmanolides, Sesquiterpenoids, Sesquiterpene lactones, Sesquiterpene acids, Flavone glycosides, Flavonoid C-glycosides, Isoflavone glycosides, Sterols, Sterol glycoside, Alkaloids, Peptide alkaloids, Amino acids, Sugars Due to research into ethnopharmacology and traditional remedies, a large number of medications have been brought to market. Ayurvedic medicine safety and practice require the establishment and validation of evidence, which will require work S.indicus Linn is one such therapeutic plant. The goal of the current investigation is to verify the medication using traditional scientific methodologies. (1)

PLANT PROFILE

PHARMACOGNOSTICAL  EVALUATION STUDIES:

MACROSCOPIC EVALUATION STUDY:

The entire Sphaeranthus indicus Linn plant was observed macroscopically. It included things like size, form, texture, color, taste, consistency, odor, and surface features.

MICROSCOPIC EVALUATION STUDY:

Using a microscope, a transverse section of Sphaeranthus indicus's leaves, roots, and stem was collected.

The properties of Sphaeranthus indicus powder were examined.     

PHYSIOCHEMICAL EVALUATION STUDIES

According to WHO criteria, the following physicochemical characteristics were determined: total ash value, sulphated ash, acid insoluble ash values, loss on drying, and sulphated ash.

DETERMINATION OF LOSS ON DRYING(LOD)

Weigh an empty, clean china dish and record its weight (W1). Weigh approximately 1.0 gram of the powdered drug into the weighed china dish. Record the combined weight of the china dish and the powdered drug (W2). Place the china dish with the drug in a hot air oven set at 105°C. Dry the sample until the weight difference between two consecutive weighings (after cooling in a desiccator) is not more than 0.5 mg. After drying, cool the china dish and sample in a desiccator to prevent moisture absorption from the air. Weigh the china dish with the dried sample and record the weight (W3) Calculate the percentage loss on drying (% LOD) with reference to the air-dried sample. (6)

FOR THE CALCULATION OF LOSS ON DRYING

As given the formula

PercentageLOD=(weight of before drying-weight of after drying)/(weight of the substance tobe taken)×100

DETERMINATION OF TOTAL ASH CONTENT:

Weigh the previously dried and weighed china dish (W1). Add the sample to the dish and weigh again (W2). Carefully heat the china dish over a small flame to char the material. This step helps in converting organic matter into carbon without losing any ash. Transfer the charred sample to a muffle furnace set at 550 ± 25°C. Maintain the temperature until a grey ash is obtained. This step ensures complete combustion of the organic matter, leaving only the ash. Cool the china dish containing the ash in a desiccator to prevent moisture Examine the ash to ensure it is carbon-free. If not, repeat the charring and ignition steps the dish from the desiccator and weigh it (W3). Repeat the heating, cooling, and weighing operations until two successive weighings do not differ significantly (within 0.001g). Record the lowest mass obtained. Calculate the Ash Content.(6)

DETERMINATION OF SULPHATED ASH :

Accurately weigh an empty, clean china dish (W1). Add 1.0 g of the powdered drug to the dish and weigh again (W2). Gently ignite the china dish until the substance is thoroughly charred, avoiding loss of ash. This step partially decomposes the organic matter. Cool the charred residue. Moisten the residue with 1 ml of sulphuric acid to help oxidize the remaining organic matter. Heat gently until white fumes are no longer evolved. This step ensures the oxidation of carbonaceous material. Place the china dish in a muffle furnace set at 800 ± 25°C. Maintain this temperature until black particles have disappeared, indicating the complete combustion of the organic material. Allow the china dish to cool. Add a few drops of sulphuric acid to the residue. Heat gently until white fumes cease and then ignite in the muffle furnace again. Allow the china dish to cool in a desiccator. Weigh the dish and record the mass (W3). Repeat the cooling, adding sulphuric acid, heating, and weighing steps until two successive weighings do not differ by more than 0.5 mg (0.0005 g). Record the lowest stable mass. Calculate the Ash Content.(6)

DETERMINATION OF ACID INSOLUBLE ASH:

The procedure  described is a typical method for determining the total ash content of a sample. Add 25 ml of dilute hydrochloric acid (HCl) to the ashed sample in the china dish. Heat the mixture on a water bath for 10 minutes. Allow the mixture to cool. Filter the contents of the dish using appropriate filter paper. Wash the filter paper with water until the washings are free from acid (you can check the pH of the washings to ensure they are neutral). Once the filter paper is free from acid, return it to the china dish. Place the dish in an oven at 100 ± 2°C for 3 hours to dry the filter paper completely. Ignite the dried filter paper in a muffle furnace at 550 ± 25°C for one hour. After igniting, cool the china dish in a desiccator to prevent moisture absorption from the air. Repeat the cooling and weighing process until two successive weighings do not differ by more than a specified amount (usually 0.1 mg). Use the recorded mass to calculate the total ash content of the sample.(6)

FOR CALCULATION OF ASH CONTENT

As given the formula

PercentageLOD=weight of before drying-weight of after dryingweight of the substance tobe taken×100

DETERMINATION OF TOTAL ASH CONTENT:

Weigh the previously dried and weighed china dish (W1). Add the sample to the dish and weigh again (W2). Carefully heat the china dish over a small flame to char the material. This step helps in converting organic matter into carbon without losing any ash. Transfer the charred sample to a muffle furnace set at 550 ± 25°C. Maintain the temperature until a grey ash is obtained. This step ensures complete combustion of the organic matter, leaving only the ash. Cool the china dish containing the ash in a desiccator to prevent moisture Examine the ash to ensure it is carbon-free. If not, repeat the charring and ignition steps the dish from the desiccator and weigh it (W3). Repeat the heating, cooling, and weighing operations until two successive weighings do not differ significantly (within 0.001g). Record the lowest mass obtained. Calculate the Ash Content.(6)

DETERMINATION OF SULPHATED ASH :          Accurately weigh an empty, clean china dish (W1). Add 1.0 g of the powdered drug to the dish and weigh again (W2). Gently ignite the china dish until the substance is thoroughly charred, avoiding loss of ash. This step partially decomposes the organic matter. Cool the charred residue. Moisten the residue with 1 ml of sulphuric acid to help oxidize the remaining organic matter. Heat gently until white fumes are no longer evolved. This step ensures the oxidation of carbonaceous material. Place the china dish in a muffle furnace set at 800 ± 25°C. Maintain this temperature until black particles have disappeared, indicating the complete combustion of the organic material. Allow the china dish to cool. Add a few drops of sulphuric acid to the residue. Heat gently until white fumes cease and then ignite in the muffle furnace again. Allow the china dish to cool in a desiccator. Weigh the dish and record the mass (W3). Repeat the cooling, adding sulphuric acid, heating, and weighing steps until two successive weighings do not differ by more than 0.5 mg (0.0005 g). Record the lowest stable mass. Calculate the Ash Content.(6)

DETERMINATION OF ACID INSOLUBLE ASH:

The procedure  described is a typical method for determining the total ash content of a sample. Add 25 ml of dilute hydrochloric acid (HCl) to the ashed sample in the china dish. Heat the mixture on a water bath for 10 minutes. Allow the mixture to cool. Filter the contents of the dish using appropriate filter paper. Wash the filter paper with water until the washings are free from acid (you can check the pH of the washings to ensure they are neutral). Once the filter paper is free from acid, return it to the china dish. Place the dish in an oven at 100 ± 2°C for 3 hours to dry the filter paper completely. Ignite the dried filter paper in a muffle furnace at 550 ± 25°C for one hour. After igniting, cool the china dish in a desiccator to prevent moisture absorption from the air. Repeat the cooling and weighing process until two successive weighings do not differ by more than a specified amount (usually 0.1 mg). Use the recorded mass to calculate the total ash content of the sample.(6)

FOR CALCULATION OF ASH CONTENT

As given the formula

Ash content=  (weight of ash)/(weight of sample taken)×100

DETERMINATION OF ALCOHOL SOLUBLE EXTRACT:

5 g of coarsely powdered, air-dried drug is macerated with 100 ml of ethanol(at the specified strength) in a closed flask.The mixture is shaken frequently for the first six hours. It is then allowed to stand for the remaining eighteen hours.The mixture is filtered rapidly to avoid loss of solvent. 25 ml of the filtrate is taken. The 25 ml of filtrate is evaporated to dryness in a pre-weighed, flat-bottomed, shallow dish.The dish with the dried extract is then dried at 105ºC until a constant weight is achieved.The final step is to weigh the dried extract and calculate the percentage of alcohol-soluble extractive with reference to the weight of the air-dried drug.(6)

DETERMINATION OF WATER SOLUBLE EXTRACT:

5 g of coarsely powdered, air-dried drug is macerated with 100 ml of chloroform water  in a closed flask. Shake the mixture frequently for the first six hours. Allow it to stand for the remaining eighteen hours. The mixture is filtered rapidly to prevent loss of solvent.25 ml of the filtrate is taken. The 25 ml of filtrate is evaporated to dryness in a pre-weighed, flat-bottomed, shallow dish.The dish with the dried extract is dried at 105ºC until a constant weight is obtained. Weigh the dried extract and calculate the percentage of chloroform-water soluble extractive with reference to the weight of the air-dried drug.(6)

FOR THE CALCULATION OF  EXTRACTIVE VALUES

As per given formula,

% of  extractive = weight of residue/weight of the drug x 100.

PHARAMACOLOGICAL  ACTIVITY

ANTICANCER ACTIVITY

Compounds:

Sphaeranthanolide, flavonoids, terpenoids

Effects:

Inhibit the growth of various cancer cell lines, including breast and colon cancer cells.(8)

 ANTI-OXIDANT ACTIVITY

Compounds:

Flavonoids (quercetin, luteolin), phenolic acid (chlorogenic acid), glycosides

Effects: 

Significant free radical scavenging activity, protecting cells from oxidative damage.(8)

ANTI-MICROBIAL ACTIVITY

Compounds:

 Extracts of Sphaeranthusindicus

Effects:

Active against a variety of bacterial and fungal pathogens, including Staphylococcus aureus and Candida albicans.(8)

 HEPATOPROTECTIVE ACTIVITY

Compounds:

Sphaerathuside, flavonoids, terpenoids

Effects:

Protect liver cells from toxin damage, improving liver function.(8)

 ANTI-DIABETIC ACTIVITY

Compounds:

Chlorogenic acid, other phenolic compounds

Effects:

Reduce blood glucose levels, improve glucose tolerance.(8)

 NEUROPROTECTIVE ACTIVITY

Compounds:

Flavonoids (quercetin, luteolin), phenolic acids

Effects:

Protect against neurodegenerative diseases by reducing oxidative stress and inflammation in neural tissues. (8)

IMMUNOMODULATORY ACTIVITY

Compounds:

Flavonoids, terpenoids

Effects:

Modulate immune responses, potentially beneficial in autoimmune conditions and immune deficiencies.(8)

ANTI-PYRETIC ACTIVITY

Compounds :

Flavonoids, other anti-inflammatory compounds

Effects :

Reduce fever in animal models comparable to standard anti-pyretic drugs.(8)

ANTI-INFLAMMATORY ACTIVITY

Compounds::

Sphaeranthanolide, flavonoids (quercetin, kaempferol), phenolic acids (caffeic acid)

Effects:

Inhibit pro-inflammatory cytokines and enzymes such as COX and LOX(8)

ANTIOXIDANT ACTIVITY

The free radical DPPH is rather stable. The evaluation of an antioxidant's capacity to scavenge the stable radical DPPH is the foundation of the experiment. Based on the principle of antioxidants, S. indicus reduces the radical to the matching hydrazine when reacting with a hydrogen donor. The solution loses color stereometrically in relation to the quantity of electrons taken up when DPPH radicals react with an appropriate reducing agent and pair off their electrons. Based on the phosphomolybednum complex formation, which was detected spectrophotometrically at 695 nm, the extract's overall antioxidant capability was determined. Flavonoids are natural products that have been demonstrated to exhibit a number of biological properties connected to the process of antioxidant action. Because of the flavonoids and other ingredients it contains, sphaeranthus indicus has antioxidant properties.(9)

IMMUNOODULATORY ACTIVITY

In tests on mice, the petroleum ether extract from the flower heads of Sphaeranthus indicus Linn. was found to be beneficial in boosting phagocytic activity, hemagglutination antibody titer, and delayed version hypersensitivity. To determine a dose-response relationship, the petroleum extract's activity was evaluated at five different dosage levels. It was discovered that the ideal dose was 200 mg/kg; at larger levels, the activity either decreased or did not rise at all. According to the current study, the medication has a lot of potential as an immunomodulatory agent because it promotes phagocytic function, humoral and cellular immunity, and both.(9)

PSYCHOTROPIC ACTIVITY

Strong neuropharmacological activity has been observed in the hydroalcoholic extract of S.indicus.The pentobarbital-induced hypnosis was extended, and spontaneous motor activity was greatly decreased by the hydroalcoholic extract of Sphaeranthus indicus. Perhaps it is acting as a light sedative. Additionally, Sphaeranthus indicus hydroalcoholic extract decreased.(9)

ANTIMICROBIAL ACTIVITY

The Sphaeranthus indicus chloroform extract has shown slightly greater efficacy than the ethanol and petroleum ether extracts against Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Klebsiella sp., Candida albicans, and Cryptococcus neoformans. Asthma, diarrhea, leucoderma, and indigestion are among the conditions for which the plant is said to be therapeutic. Researchers extracted a novel sesquiterpene glycoside, sphaeranthanolide, and isoflavone glycoside from S. indicus leaves and flowers. These compounds were discovered to be immunological stimulants. Treatments for skin diseases, coughs, and fevers include S.indicus, according to medical knowledge from tribal healers. Pile treatment is reportedly effective when the bark is crushed and combined with whey. Thin layer chromatographic separation produced the active constituent 7 hydroxyfrullanolide, and the chloroform extract of leaves in this investigation demonstrated strong action against the chosen human infections. This substance, which is classified as sesquiterpenes, was extracted from a floral portion of Spaeranthus indicus. It is possible to analyze this herb's composition for the treatment of many ailments, such as cancer and skin conditions, and to create a herbal medication since it has a wide spectrum of therapeutic value.(9)

ANTIDIABETIC ACTIVITY

In rats administered streptozotocin (STZ) (60 mg?kg?^(-1 ) i.p.) and nicotinamide (120 mg?kg?^(-1)) to induce diabetes, we examined the potential antihyperglycemic effects of Sphaeranthus indicus extract. Normal and diabetic rats were used to assess fasting plasma glucose levels, serum insulin levels, serum lipid profiles, magnesium levels, glycosylated hemoglobin, changes in body weight, and liver glycogen levels. After taking S. indicus orally for 15 days, there was a notable drop in blood glucose levels and an increase in liver glycogen and plasma insulin. When given the alcoholic extract of S. indica, rats that were fasting and not diabetics showed a notable improvement in the oral glucose tolerance test. The reference standard that was employed was glibenclamide. The results show that Sphaeranthus indicus extract, which is alcoholic, may have potential benefits in the management of diabetes.(9)

ANTIHYPERLIPIDEMIC ACTIVITY

Serum TC and LDL-c levels increased, but cholesterol levels decreased. The animal fed an atherogenic diet had elevated HDL levels. The main risk factor for coronary heart disease (CHD) was elevated blood cholesterol, with HDL serving as a cardioprotective lipoprotein. The levels of TC and LDL-c are dramatically reduced by treatment with 500 mg/kg/day of Sphaeranthus indicus extract. Lecithin's increased activity might be the cause of the Sphaeranthus indicus extract's effects. The TG level decreased when Sphaeranthus indicus extract (500 mg/kg/day) was administered. LDL:HDL-c ratio and atherogenic index were significantly lower. Both high and low blood cholesterol levels may be predicted by the LDL:HDL-c ratio, and the most significant marker of coronary heart disease (CHD) is the atherogenic index. Strong antihyperlipidemic effects are demonstrated by the alcoholic extract of Sphaeranthus indicus. (9)

ANXIOLYTIC ACTIVITY

The animals that were given extracts or diazepam (1 mg/kg) displayed increased walking, activity in the center of the test, and total locomotion; they also showed decreased bouts of fighting in the foot-shot-induced aggression, which suggested anxiolytic activity; and finally, they showed an increase in the time spent, percent entries, and total entries in the open arm of the elevated pulse maze (EPM). Three extracts from the flowers of Sphaeranthus indicus produced significant activity in the mice: petroleum ether (10 mg/kg), alcoholic (10 mg/kg), and water (30 mg/kg). The EPM and OFT showed a greater anxiolytic effect from petroleum ether extract (10 mg/kg) compared to ethanolic and water extracts; however, it was not as strong as that of diazepam (1 mg/kg).(9)

PHARMACOGNOSTICAL EVALUATION STUDIES:

The primary and most trustworthy criterion for identifying plant-based medications is the pharmacognostic analysis. The pharmacognostical characteristics are required for identification verification as well as for assessing the crude drug's quality and purity. The comprehensive and methodical pharmacognostic assessment would provide important data for subsequent research.

TRANSVERSE SECTION OF LEAF

The leaf is dorsiventral and exhibits an extensive number of various sorts of trichomes on both the epidermis. Simple trichomes have thick walls, three to four cells, with dimensions of 130.8–145.2 ?m in length and 29.0–43.5 ?m in breadth. Trichomes have a straight or knee-shaped shape, a swelling base, and a collapsed cell in the center or at the tip. Three or four collateral vascular bundles connected to a cluster of sclerenchymatous cells on either side can be seen in the midrib.

A discontinuous ring of lignified pericyclic fibers and a well-developed ring of bicollateral vascular bundle around the pith are characteristics of the stem that indicate cork with two to three layers of parenchymatous cells covered with papillose cuticle possessing trichomes. Pitted and lignified, medullary rays are approximately unitetraseriate.

On its exterior surface, the root displays the characteristic brown tissue known as the metaderm. Forming an uneven layer, it is made up of suberized cells that act as a barrier. The secondary cortex has radially distributed secretory canals that alternate with radial groupings of pericyclic fibers and a few stone cells. Radially oriented parenchymatous phloem is present. Two to five seriate, lignified, and pitted medullary rays are present.

1. Habit
2. Flowering Twig
3. Crude Drug
4. T.S Of Leaf Through Midrib
5. Stele Enlarged
6. Vascular Bundle
7. Portion Showing Trichomes
8. Portion Of Lamina Showing Palisade And Spongy Parenchyma
9. Venation Pattern
10. Stomata Enlarged
11. T.S. Of Stem Showing Various Regions
12. Portion Showing Epidermis And Cortex
13. Portion Showing Vascular Bundle
14. Pith Cells
15. Druse Type Crystal In Pith Cells
16. Pitted Parenchyma
17. T.S Of Root Showing Various Regions
18. Portion Showing Cork And Sec. Cortex
19. Secondary Cortex Showing Oil Globules
20. Secondary Phloem Showing Oil Globules
21. Secondary Xylem Showing Vessels And Medullary Rays
22. Medullary Rays With Pits

25-26. Parenchyma Cells

27. Group Of Fibres

28. Fibre With Forked End

 29. Fibre With Drawn Out End

 30. Tracheid

 31. Pitted Parenchyma

 32. Vessel With Spiral Thickenings

33-35. Vessels With Pitted Thickenings

 36-37. Vessels With Terminal And Sub Terminal Opening

 Powder Characters

 38-39. Multicellular Covering Trichomes

40. Glandular Trichome

 41. Cork Cells

 42. Parenchyma Cells

 43. Vessel With Spiral Thickenings

 44. Vessel With Simple Pits

 45. Vessel With Scalariform Thickenings

 46. Vessel With Bordered Pits

 47. Druse Type Of Crystals

PHYSIOIOCHEMICAL EVALUATION STUDIES

Standards for crude pharmaceuticals can be set with the use of the quantitative determination of some pharmacognostical factors. In order to identify drug adulteration or inappropriate management, it is crucial to continuously evaluate the drugs physically. The medication may inhibit the growth of bacteria, fungi, or yeast because its moisture level is not excessively high. Determining the ash value and acid-insoluble ash value is equally significant in the assessment of crude medicines. Total ash, or the presence or absence of foreign inorganic materials such metallic salts and/or silica, is particularly significant for evaluating the purity of pharmaceuticals. Sphaeranthus indicus Linn crude powder analysis physicochemical parameter data are shown in the table.

Sphaeranthus indicus Linn.,

         Alkaloids, flavonoids, sugars, proteins, tannins, steroids, and saponin glycosides were found, according to a preliminary screening of phytochemicals.