INTRODUCTION:

Inflammation and pain are protective responses but, when persistent, contribute to chronic diseases such as arthritis and neuropathic pain. Conventional treatment relies heavily on NSAIDs and opioids, which provide symptomatic relief but carry risks of gastrointestinal bleeding, cardiovascular complications, renal impairment, and tolerance or dependence in long-term use. This necessitates the search for safer and effective alternatives.^1,2,3

Ayurveda, a 5,000-year-old comprehensive healthcare system, describes numerous plant-based formulations with analgesic and anti-inflammatory properties.⁴ Several medicinal plants rich in flavonoids, alkaloids, tannins, and glycosides have been reported to modulate inflammatory mediators and nociceptive pathways.^5,6 Previous studies have demonstrated the potential of herbal formulations such as polyherbal oils, decoctions, and tablets in preclinical models of inflammation and pain. However, systematic comparative evaluation of marketed Ayurvedic formulations remains limited.⁷

Analgesics are medications that work on the central or peripheral neural systems to reduce pain without substantially changing consciousness. They do not completely block feeling, in contrast to anesthetics. NSAIDs (like ibuprofen) and opioids (like morphine) are common varieties.⁸ Although NSAIDs are frequently usedto relieve pain and inflammation, they can also have negative side effects, such as cardiovascular problems, renal impairment, and stomach ulcers. Therefore, safer and more effective treatments are still needed, particularly for chronic illnesses.^9,10

Using proven in vivo models, including carrageenan-induced paw edema, the hot plate test, the tail immersion method along with writhing in rodents due to acetic acid, current study aimed to assess and contrast an anti-inflammatory as well as analgesic capabilities of Rhumayog Tablet (RT) and Sandhivatari Tablet (ST). The study is to estimate the usefulness of these Ayurvedic formulations in comparison to common NSAIDs such as diclofenac sodium and indomethacin, as well as to offer scientific support for their traditional use.

MATERIAL AND METHODS:

RT and ST were obtained from local market. RT, Marketed by Zandu (Batch No.: EO0002) and ST, Marketed by Sanjivani Pharma (Batch No.: 35). Indomethacin and diclofenac sodium were used as standard reference drugs.

DOSE SELECTION:

The doses for RT (350mg/kg) and ST (250mg/kg) were calculatedby converting the human therapeutic dose to animal equivalent doses based onbody surface, area, in accordance with standard guidelines. Pilot studies and previous literature on similar Ayurvedic formulations further supported the selected doses.

Animals:

Healthy Wistar rats (150–250g) and the Swiss albino mice (20–30gm), of either sex, were obtained from the National Institute of Biosciences, Nigwewadi, Maharashtra. Rats were housed in groups of six and mice in groups of five under controlled temperature (27 ± 2°C), 12h light/dark cycle and free.access for food and water. All procedures were conducted with prior approval ofthe Institutional Animal Ethics Committee (IAEC/AMCP/01/2024-25) and in compliance with CPCSEA guidelines. The experimental animal study was conducted inthe animal house in pharmacology department, Ashokrao Mane College of Pharmacy, Peth Vadgaon (Registration Number- 1232/PO/Re/S/08/CPCSEA). The number of animals used was the minimum required to achieve statistical significance. Humane endpoints were predefined to minimize suffering, and animals were provided with adequate housing, nutrition, and care throughout the study.

Phytochemical Investigation:

Preliminary phytochemical screening of RT and ST was conducted according to standard protocols described by K.R. Khandelwal.¹¹ Both formulations tested positive for tannins, phenols, alkaloids, flavonoids, glycosides, carbohydrates, steroids, and saponins.

Although qualitative phytochemical detection was performed, future work should focus on quantifying marker constituents (e.g., total flavonoid content or HPLC fingerprinting) to ensure reproducibility and batch-to-batch consistency.

Anti-inflammatory Activity¹²

The carrageenan-induced ratpaw edema model was used in Wistar rats (n = 6 per group). Animals were separated into five groups:

Group I: Normal control

Group II: Induced (carrageenan, 0.1Ml of 1% w/v solution, sub-plantar injection)

Group III: Standard, (Indomethacin, 10mg/kg, p.o.)

Group IV: RT (350mg/kg, p.o.)

Group V: ST (250mg/kg, p.o.)

Drugs were given orally one time daily for three serial days prior to carrageenan injection. On third day, carrageenan (0.1 milliliter of the 1%) was injected to sub-plantar area of each rat’s left hind paw after 30 min of the last drug dose. The volume of Paw was measured, at 0, 1, 2, 3, 4, and 5hrs with the help of a digital plethysmometer. The formula given below was used to determine the percentage of reduction in inflammation:

Percentage (%) Inhibition = Vc-Vt/Vc × 100

Where, Vc denotes mean. paw volume for control and Vt = mean paw volume of treated groups.

Analgesic Activity:

A number of methods were employed to evaluate analgesic activity, such as hot plate and tail immersion along with writhing caused by acetic acid induction. Total 20mice were chosen at random, allotted to Four distinct groups, (n= 5 per group). Group I (Control), while the second group was given Diclofenac sodium with distilled water orally (50mg/kg) animal body weight as the Standard. Third and fourth group was taken as test group which receives test tablets RT at low and also high dose (250mg/kg ; 350mg/kg, by oral route) and ST at low as well as high dose (150mg/kg ; 250mg/kg, by oral route). All the animals were reused for analgesic models after the washout period.

Analgesia Meter- Hot Plate¹³

Prior to the treatment each animal was placed separately into hot plate that was sustained at an ambient temp. of 55±2°C and the measurement of its reaction time considered as initial reaction. Once the initial response time has been measured, the treatment such as RT, ST as well as reference standard drug i.e. Diclofenac sodium (50mg/kg) was given at different selected dosage to each mouse and then placed individually on an Eddy's hot plate that kept at a controlled temp., and a paddle switch was used to record the hot plate a latency period while the forepaws were licking. After the standard as well as test drug administration the response time was recalculated for 30,60,90 and 120 minutes.

Tail Immersion¹⁴

Mice in test and also standard group were treated with selected Ayurvedic formulations (RT, ST) at different selected dosage and Diclofenac sodium (50mg/kg; orally). After dosing the last 2 to 3cm section of every mouse tail was marked and remained submerged in warm water that was kept on 55±0.5ºC. The reaction time for each mice was measured by the mouse's time to take its tail out of the hot water. After the standard and test drugs were administered orally for 0, 15, 30, 45, 60, and 120 minutes, the reaction time was recalculated.

Acetic Acid Induced Writhing¹⁵

One hour after treatment (Diclofenac sodium, RT, ST), animals received ip inj.of acetic acid (0.6%, 10mL/kg). To evaluate analgesic effect of test tablets, the no. of writhes brought on due to 0.6 percent acetic acid (10 ml/kg, Intraperitoneal) in next 20minutes, were recorded.

Percentage inhibition was calculated to compare the analgesic activity of R.T. as well as S.T. with the standard analgesic drug that is Diclofenac sodium. It is calculated by formula:

Percentage Inhibition = Wc - Wt/ Wc × 100

Were, Wc represents mean number of writhes in control and Wt = mean number of writhes in treated groups.

Statistics:

The mean±standard error ofmean, (S.E.M.) were employed to represent all outcomes. One-way analysis ofvariance (ANOVA) performed by Dunnett's multiple comparison test, were used regarding statistical comparisons. p-values lower than 0.05, 0.01, or 0.001 were considered as statistically significant.

RESULTS:

Carrageenan Induced Rat Paw Edema:

The outcomes concise in Table 1 have exposed an anti-inflammatory action of both RT and ST. Carrageenan injection produced a progressive increase in paw volume in the induced group. Both RT (350mg/kg) and ST (250 mg/kg) significantly (p<0.001) reduced paw edema compared with the induced group. RT showed 23.7% inhibition, while ST demonstrated 23.1% inhibition at 5 hours, values comparable to indomethacin (28.4%). These findings confirm the anti-inflammatory activity of both formulations, with RT displaying slightly greater efficacy. (Graph no.1)

Table 1: Effect of Rhumayog Tablet (350 mg/kg) Sandhivatari Tablet (250 mg/kg) on Carrageenan Induced Paw Edema

Groups	Dose (mg/kg)	Rat Paw Edema Volume (in ml)						Percentage Inhibition at 5hr
Groups	Dose (mg/kg)	0 hr	1 hr	2hr	3hr	4hr	5 hr	Percentage Inhibition at 5hr
NORMAL	-	1.065± 0.02513	1.113± 0.02186	1.082± 0.01973	1.067± 0.04224	1.072± 0.02344	1.085± 0.02884	-
INDUCED (Carrageenan)	0.1ml of 1% W/V	1.342± 0.05552***	1.443± 0.04681***	1.512± 0.04757***	1.638± 0.06074***	1.747± 0.05795***	1.695± 0.05755***	-56.22
Indomethacin	10 mg/kg	1.283± 0.03373	1.382± 0.02822	1.455± 0.02884	1.422± 0.03260**	1.372± 0.02915***	1.293± 0.03461***	28.40
Rhumayog Tablet	350 mg/kg	1.19± 0.03141*	1.258± 0.02455**	1.323± 0.02201***	1.34± 0.01633***	1.303± 0.02028***	1.21± 0.01949***	23.66
Sandhivatari Tablet	250 mg/kg	1.23± 0.02569	1.312± 0.02822*	1.387± 0.02431*	1.438± 0.02182**	1.395± 0.03274***	1.303± 0.02963***	23.07

The observations are mean± S.E.M., *p<0.05: Less significant, **p<0.01: significant, ***p<0.001 Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Graph No. 1: Graphical representation of Effect of Rhumayog Tablet (350 mg/kg) Sandhivatari Tablet (250 mg/kg) on Carrageenan Induced Rat Paw Edema at different time intervals.

Table 2: Effect of Rhumayog Tablet (250 and 350 mg/kg) on Hot plate method in mice

Groups	Dose (mg/kg)	Basal Reaction Time (sec)
Groups	Dose (mg/kg)	0 min	30 min	60 min	90 min	120 min
CONTROL	-	6.2± 0.3742	6.4± 0.5099	7± 0.3162	7± 0.3162	8± 0.3162
Diclofenac Sodium	50 mg/kg	6.6± 0.5099	9± 0.3162***	11.6± 0.2449***	13± 0.3162***	14.6± 0.2449***
Rhumayog Tablet	250 mg/kg	5.8± 0.2000	7.6± 0.4000	9.2± 0.4899**	10.8± 0.3742***	12.4± 0.5099***
Rhumayog Tablet	350 mg/kg	6.4± 0.4000	8.8± 0.3742**	11± 0.3162***	12.8± 0.3742***	13.6± 0.4000***

The observations are mean± S.E.M., **p<0.01: significant, ***p<0.001 Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Graph No. 3: Graphical representation of Effect of Sandhivatri Tablet (150 and 250 mg/kg) on Hot plate method in mice at different time intervals.

Graph No. 2: Graphical representation of Effect of Rhumayog Tablet (250 and 350 mg/kg) on Hot plate method in mice at different time intervals.

Table 3: Effect of Sandhivatri Tablet (150 and 250 mg/kg) on Hot plate method in mice

Groups	Dose (mg/kg)	Basal Reaction Time (sec)
Groups	Dose (mg/kg)	0 min	30 min	60 min	90 min	120 min
Control	-	6.2± 0.3742	6.4± 0.5099	7± 0.3162	7± 0.3162	8± 0.3162
Diclofenac Sodium	50 mg/kg	6.6± 0.5099	9± 0.3162***	11.6± 0.2449***	13± 0.3162***	14.6± 0.2449***
Sandhivatari Tablet	150 mg/kg	5.6± 0.4000	7.8± 0.2000*	10.2± 0.3742***	11.8± 0.5831***	13± 0.5477***
Sandhivatari Tablet	250 mg/kg	6.4± 0.2449	8.6± 0.2449***	11± 0.3162***	12.8± 0.4899***	14.2± 0.3742***

The observations are mean± S.E.M., *p<0.05: Less significant, ***p<0.001Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Table 4: Effect of Rhumayog Tablet (250 and 350 mg/kg) on Tail Immersion method

Groups	Dose (mg/kg)	Latency to flick tail (sec)
Groups	Dose (mg/kg)	0 min	15 min	30 min	45 min	60 min	120 min
Control	-	2.742± 0.1975	2.896± 0.03027	3.022± 0.1210	3.02± 0.08155	2.984± 0.04697	3.024± 0.05980
Diclofenac Sodium	50 mg/kg	3.206± 0.2393	3.676± 0.4080	3.86± 0.4073	4.336± 0.4186**	4.67± 0.4085***	4.324± 0.3763**
RT	250 mg/kg	2.934± 0.08004	3.238± 0.1126	3.48± 0.1201	3.74± 0.1399	4.086± 0.1340**	3.826± 0.08501*
RT	350 mg/kg	3± 0.08994	3.294± 0.1278	3.578± 0.1267	3.928± 0.1649*	4.272± 0.1546**	4.04± 0.1664*

The observations are mean± S.E.M., *p<0.05: Less significant, **p<0.01: significant, ***p<0.001 Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Graph No. 4: Graphical representation of Effect of Rhumayog Tablet (250 and 350 mg/kg) Tail Immersion method at different time intervals.

Table 5: Effect of Sandhivatari Tablet (150 and 250 mg/kg) on Tail Immersion method

Groups	Dose (mg/kg)	Latency to flick tail (sec)
Groups	Dose (mg/kg)	0 min	15 min	30 min	45 min	60 min	120 min
CONTROL	-	2.742± 0.1975	2.896± 0.03027	3.022± 0.1210	3.02± 0.08155	2.984± 0.04697	3.024± 0.05980
Diclofenac Sodium	50 mg/kg	3.206± 0.2393	3.676± 0.4080	3.86± 0.4073	4.336± 0.4186**	4.67± 0.4085***	4.324± 0.3763**
ST	150 mg/kg	3.004± 0.1997	3.184± 0.1693	3.62± 0.1811	3.866± 0.2031	4.166± 0.1712**	3.864± 0.1893
ST	250 mg/kg	3.114± 0.2055	3.364± 0.2238	3.7± 0.2540	3.998± 0.2033*	4.328± 0.1903**	3.942± 0.2033*

The observations are mean± S.E.M., *p<0.05: Less significant, **p<0.01: significant, ***p<0.001 Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Graph No. 5: Graphical representation of Effect of Sandhivatari Tablet (150 and 250 mg/kg) on Tail Immersion method at different time intervals.

Acetic Acid Induced Writhing:

In contrast to acetic acid, RT (250 & 350 mg/kg) significantly lowered the amount of writhings. By RT (350 mg/kg), the greatest percentage suppression of the writhing response was seen. Diclofenac sodium exhibited a maximal writhing response inhibition of 67.49%. The observations are given in table no. 6.

Table 6: Effect of Rhumayog Tablet (250 and 350 mg/kg) on Acetic acid induced writhing method

Groups	Dose	No. of writhes	Percentage Inhibition
CONTROL	-	56.6 ± 1.208	-
Diclofenac Sodium	50 mg/kg	18.4 ± 1.208***	67.49
R.T.	250 mg/kg	35.8 ± 2.223***	36.74
R.T.	350 mg/kg	21 ± 1.304***	62.89

The observations are mean± S.E.M., ***p<0.001 Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Acetic acid-induced writhing is considerably reduced by ST (150 and 250 mg/kg). ST (250 mg/kg) exhibited the greatest percentage of writhing response reduction. The highest level of writhing response inhibition demonstrated by diclofenac sodium was 67.49%. Table No. 7 provides the observations.

Table 7: Effect of Sandhivatari Tablet (150 and 250 mg/kg) on Acetic acid induced writhing method

Groups	Dose	No. of writhes	Percentage Inhibition
Control	-	56.6 ± 1.208	-
Diclofenac Sodium	50 mg/kg	18.4 ± 1.208***	67.49
S.T.	150 mg/kg	41.80 ± 2.354***	26.14
S.T.	250 mg/kg	26.20 ± 1.463***	53.71

The observations are mean± S.E.M., ***p<0.001 Extremely significant (One Way ANOVA followed by Dunnet’s Test)

Graph No. 6: Graphical representation of Effect of (a) Rhumayog Tablet (250 and 350 mg/kg) as well as (b) Sandhivatari Tablet (150 and 250 mg/kg) on Acetic acid induced writhing method

DISCUSSION:

Numerous Ayurvedic therapeutic preparations comprising oils, minerals, and calcined metals in combination with herbs are extensively utilized in traditional medicine systems for their capability to decrease inflammation as well as pain. These preparations are widely marketed within India and exported internationally, primarily for therapy for inflammatory diseases including arthritis. However, there exists a lack of systemic, protocol-based investigations substantiating their pharmacological efficacy. Moreover, comprehensive reports on their chemical characterization and biological activities remain limited.^[15,16-20] Hence, there is a compelling need for scientific validation of these formulations, which are based on ancient Ayurvedic knowledge.

The current investigation offers experimental proof that the anti-inflammatory and analgesic properties of two Ayurvedic formulations, Rhumayog Tablet (RT) and Sandhivatari Tablet (ST). Both demonstrated significant activity across validated models of inflammation and pain, with RT showing slightly greater efficacy than ST.

In the carrageenan-induced paw edema model, RT and ST reduced paw swelling by approximately 24% and 23%, respectively, compared with 28% inhibition by indomethacin. Since this model is mediated initially by histamine and serotonin, followed by prostaglandins and cytokines, the findings suggest that both formulations may interfere with multiple mediators of inflammation. However, the precise mechanisms remain unclear and require further molecular studies.

Analgesic evaluation in central (hot plate and tail immersion) and peripheral (acetic acid writhing) models demonstrated significant dose-dependent effects. RT (350 mg/kg) provided analgesia comparable to diclofenac sodium in both thermal and chemical nociception assays, while ST was moderately less potent. These results indicate that the formulations may modulate both central and peripheral pain pathways, possibly through cyclooxygenase inhibition or suppression of inflammatory mediators, though this remains speculative.

This study provides pharmacological evidence supporting the use of RT and ST in musculoskeletal pain and inflammatory conditions. RT appears slightly more potent, suggesting potential clinical preference. Future research should focus on: Standardization of formulations using marker compounds, Mechanistic studies on inflammatory mediators and signaling pathways, Long-term safety and toxicity evaluations, controlled clinical trials to confirm efficacy in patients.

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Received on 30.06.2025 Revised on 06.11.2025

Accepted on 08.01.2026 Published on 01.07.2026

Available online from July 04, 2026

Research J. Pharmacy and Technology. 2026;19(7):2999-3006.

DOI: 10.52711/0974-360X.2026.00427

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.