PHYTOCHEMICAL CHARACTERIZATION OF AN AYURVEDIC FORMULATION TRIJATAKADI GUTIKA USING GC-MS WITH POSSIBLE ACTION ON KANTAROGA (THROAT DISEASES)

Standardisation of polyherbal formulations plays a key role in estimating the quality and safety of drugs. In this study, a novel polyherbal formulation was prepared for the treatment of diabetic nephropathy and it was attempted to evaluate for its safety and efficacy against free radicals. To ascertain the safety of the formulation, individual herbs and polyherbal formulation have been screened for microbial load, heavy metals, pesticide residues and aflatoxins according to Ayush guidelines. Phytochemical analysis was carried out using standard procedures and GCMS analysis to identify the bioactive compounds. Antioxidant methods such as DPPH, nitric oxide, hydrogen peroxide, hydroxyl and superoxide radical scavenging assays were undertaken. To predict the mechanism behind the antioxidant efficacy, a molecular docking analysis was performed against Keap1/Nrf2 (PDB ID: 2FLU) protein using AutoDock Vina software. Phytochemical analysis showed the presence of alkaloids, tannins, flavones, glycosides, polyphenols, phytosterols and saponins. The results of in vitro antioxidant studies showed increase in percentage inhibition with increase in concentration and found no significant difference compare with standards. Among the 15 bioactive compounds identified from GCMS analysis, guanosine showed high binding affinity with Keap1/Nrf2 and interacting with similar residues as that of standard ascorbic acid which is analysed through molecular docking. Based on the results, it has been concluded that the safety parameters of polyherbal formulation were found within specified limits of Ayush guidelines and its efficacy against oxidative stress plays an effective role to treat diabetic nephropathy due to its synergistic antioxidant effect.

Polyherbalism, an alternative natural-based therapy for various disorders, has been quoted about 1,300 years before in Sharangdhar Samhita. Herbal-based combination therapy stages a vital role for the treatment of type 2 diabetes mellitus (T2DM) and associated complications. The present study aims at developing an Ayurvedic-based polyherbal formulation (ADPHF6) and the assessing its antidiabetic and antioxidant property. ADPHF6 polyherbal formulation was measured for phytochemical components by qualitative methods. The polyherbal formulation was quantitatively estimated for its phytochemical constituents, i. e. total phenol and flavonoid content. Further, the antioxidant property of ADPHF6 formulation was evaluated by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging assay, hydrogen peroxide radical scavenging assay and metal chelating assay. α-Amylase and α-glucosidase inhibitory activities of polyherbal formulation were also assessed. ADPHF6 was further analysed for its protective antioxidant property against reactive oxygen species (ROS‾)-induced damage in human lymphocyte DNA and pUC19 plasmid. ADPHF6 polyherbal formulation revealed the presence of phytochemical constituents such as alkaloids, flavonoids, phenols, tannins, terpenoids, saponins and cardiac glycosides in significant levels. Further, it also measured the higher levels of total phenols (473.3±3.05 mg/g) and flavonoid (664±5.29 mg/g) content. Polyherbal formulation also exhibited IC50 values of 49.9±0.15, 65.1±0.10 and 60.1±0.05 mg/mL for 2,2- diphenyl-1-picryl-hydrazyl-hydrate (DPPH), hydrogen peroxide (H2O2) and Fe2+ radical scavenging activities, respectively. ADPHF6 revealed an inhibitory activity (IC50) of 0.67±0.01 and 0.81±0.01 mg/mL for α-amylase and glucosidase, respectively. Pre-treated human peripheral blood lymphocytes with ADPHF6 aqueous extract illustrated enhanced protection against ROS-mediated damage as compared with post-treated groups. DNA nicking assay rendered protective activity against the OH¯ radical-induced DNA damage in supercoiled pUC19 plasmid. Our present study demonstrates that ADPHF6 offers potent inhibitory activity against free radicals as well as digestive enzymes. However, studies should be conducted using in vivo model to further elucidate the effect against free radicals and its anti-hyperglycaemic activity in the management of non-insulin-dependent diabetes.

A polyherbal medicine, Habb-e-Ustukhuddus (HU), is used for its anti-inflammatory properties. However, the anticancer and chemopreventive properties of HU were not known, and Therefore, investigated in the present study. Cancer cells were treated with 50-400 µg/ml HU and MTT, trypan blue, and clonogenic assays were performed. Propidium iodide (PI) staining, annexin V-FITC assay, and JC-1 staining were done for cell cycle progression, apoptosis, and mitochondrial membrane potential, respectively, using flow cytometry. Immunoblotting, cell migration and invasion assays were performed. Chemical characterization of HU was done through GC-MS and HPLC analyses. C57BL/6 mice were used to assess the in vivo toxicity of HU. While evaluating the anticancer activity, the methanolic extract of HU (50-400 µg/ml) strongly inhibited the growth and survival (P<0.05-0.001) of lung and breast cancer cells and increased the cell population in the sub-G1 phase of the cell cycle. HU caused apoptotic death of cancer cells (P<0.05-0.001), which was associated with the depolarization of mitochondrial membrane potential (Δψ) (P<0.001) and an increase in Bax to Bcl-2 protein ratio. Further, HU inhibited the invasion and migration of cancer cells, which was accompanied by an increase in the epithelial marker, E-cadherin, and a decrease in the mesenchymal marker, vimentin. The HU characterization by GC-MS and HPLC analyses showed the abundance of bioactive compounds including flavonoids and alkaloids. In the chemopreventive study, the oral administration of methanolic extract of the formulation HU (50 and 100 mg/kg body weight) to mice did not cause any toxicity and significantly increased the specific activities of hepatic drug metabolizing phase I and phase II enzymes, which suggested for its detoxification potential of xenobiotic compounds. Together, these results demonstrated the anticancer potential HU, without any apparent toxicity in mice, and thus HU could be further explored for its clinical utility in cancer control.

Biofilm is a complex microbial community highly resistant to antimicrobials. The formation of biofilms in biotic and abiotic surfaces is associated with high rates of morbidity and mortality in hospitalized patients. The aim of the present study was to develop a polyherbal formulation for antibacterial activity and inhibition of biofilm formation. Andrographis paniculata (leaves), Cassia fistula (flower), Gymnema sylvestre (leaves), Ocimum tenuiflorforum (leaves), Cinnamomum verum (bark)were the herbs chosen for the study. The extraction of the herbal samples was carried out using the cold extraction method and their phytochemical characterization was carried out by GC-MS analysis. The anti-microbial activity and antibiofilm activity of polyherbal formulation was carried out using the agar well diffusion method and microtitre plate assay. The phytochemical results showed the presence of resins, carbohydrates, proteins, saponin and phenol. The agar well diffusion assay results and microtitre plate assay showed the highest activity exhibited with 500µg/ml of polyherbal extract against Pseudomonas aeruginosa and lowest against Escherichia coli. The results confirm the differences in antibacterial and antibiofilm activity of polyherbal formulation of various concentration. Larger concentration of formulation showed to be more effective against biofilm cells. The purpose of this study was to investigate the anti-biofilm properties and inhibitory impact of ACGOC extracts against the most prevalent clinical isolates such as S. aureus,B.pumilusand P. aeruginosa and so the hypothesis of synergistic interaction of biologically active compounds from five different herbal extracts was proved for its antibiofilm assays.

Background: Novel polyherbal formulation (PHF) is the utilization of more than one herb in the preparation of herbal medication. The thought is found in the conventional system of medicine where the variety of herbs in a specific proportion of illness. Because of synergism, polyherbalism presents a few advantages which aren't accessible in single herbal medication. It is utilized in these medications for the treatment of numerous sicknesses including antioxidants. Objective: To develop a phytochemical screening and GC-MS analysis of Novel Polyherbal formulation for In vitro antioxidant activity. Materials and Methods: Macroscopical, preliminary phytochemical, quantitative phytoconstituents, and In-vitro antioxidant activity of all the individual extract and polyherbal formulation was done by chemical method. Identification of phytoconstituents with the aid of Gas chromatography – Mass spectroscopy (GC-MS). Results: Macroscopical study and physicochemical examination, for example, ash value, extractive value, loss on drying, and pH were reported to A. racemosus, B. variegata, C. bonducella, S. asoka, and S. racemosus and novel polyherbal formulation. Qualitative phytochemical investigation revealed the presence of alkaloids, flavonoids, gums & mucilage, carbohydrates, steroids, proteins & amino acids, fats & fixed oils, glycoside, phenols, and saponins. Quantitative estimation such as TAC, TFC, TGC, TSC, and TPC was showed positive results. All the individual extract and PHF were subjected to GC-MS analysis. All the individual extract and polyherbal formulation displayed strong antioxidant activity. Conclusions: To conclude the PHF was reported that high level of bioactive contents present and strong antioxidant activity in contrast to the preferred ascorbic acid. The GC-MS uncovered the presence of bioactive compounds and these compounds are suggested to treat antibacterial, antioxidant, anti-inflammatory, and antiviral, anti-tumor, anti-proliferative activity, and antifungal activity.

Combinations of many medicinal plants, called polyherbal formulations, have been used to make medicines work better because the bioactive compounds in them work together more effectively. Consequently, The aim of this paper is to assess the phytochemical compounds and analgesic properties of poly-herbal tea formulated from Anthocleista djalonensis, Musa paradisiaca, Mucuna pruriens, Thespesiagarckeana, and Curcuma longa to decrease induced pain on adult albino mice using appropriate standard methods. Data obtained show that the polyherbal formulated tea (Anthocleista djalonensis, Musa paradisiaca, Mucuna pruriens, Thespesiagarckeana and Curcuma longa) contains 17 compounds identified by GC-MS analysis, and they include: 2-(3,7-Dimethyl-octa-2,6-dienyl) -4-methoxyphenol, 2-(3,7-Dimethyl-octa-2,6-dienyl) -4-methoxyphenol, 2-Methoxy-4-m Benzene, 1-(1,5-dimethyl-4-hexenyl) -4-methyl, 1,3-Cyclohexadiene, 5-(1,5-dimethyl-4-hexenyl)-4-m -6-methylene-, Ledol, Tumerone, 3-Benzofuranmethanol, 2,3-dihydro-2-(4-hydroxy-3-methoxyphenyl), Curlone, Inositol, 1-deoxy-, alpha-D-mannofuranoside, methyl, carinol, 2(3H)-furanone, dihydro-3,4-bis[(4-hydroxy-3-methoxyphenyl)methyl] -, (3R-trans)-, Phenethyl alcohol, 2,5-dihydroxy-.alpha.-methyl- (-), 2-Methyl-4-octenal. Results obtained reveal that analgesic property of the polyherbal formulated tea could possess pain-relieving effects similar to that of pentazocine and aspirin, suggesting that the poly-herbal formulated tea may work by interacting with opioid receptors or by inhibiting cyclooxygenase enzymes. Overall, this study supports the use of these plants mixture in traditional medicine for the treatment of pain.

Enprostil is a synthetic prostaglandin E2 analogue with gastric antisecretory and mucosal protective properties. We compared the effects of enprostil and ranitidine on the healing of gastric ulcers and the subsequent relapse rates over 6 months. Patients (N = 156) were recruited for a double-blind study from 12 centers in Europe; 71 were randomly assigned to oral treatment with 35 micrograms enprostil twice daily and 85 to 150 mg ranitidine twice daily for up to 8 weeks. Both groups were of similar demography; their healing rates were also similar. Cumulative intent-to-treat healing rates were at 4 weeks enprostil 48%, ranitidine 41%: at 6 weeks enprostil 65%, ranitidine 68%; and at 8 weeks enprostil 72%, ranitidine 80%. Of those patients who met all protocol criteria and completed treatment, and were endoscoped at the prescribed times, healing rates were at 4 weeks enprostil 55%, ranitidine 54%, at 6 weeks enprostil 75%, ranitidine 84%; and at 8 weeks enprostil 80%, ranitidine 90%. Relief of pain was rapid and similar in both groups. The incidence of adverse events was low and similar in the two groups. The treatment-free relapse rate at 6 months was enprostil 64%, ranitidine 49%; the median times to relapse were 169 and 203 days, respectively. Enprostil and ranitidine appear to be equally effective in healing gastric ulcers.

1. JB-9322 is a selective histamine H2-receptor antagonist with gastric antisecretory activity and mucosal protective properties. 2. The affinity of JB-9322 for the guinea-pig atria histamine H2-receptor was approximately 2 times greater than that of ranitidine. 3. In vivo, the ID50 value for the inhibition of gastric acid secretion in pylorus-ligated rats was 5.28 mg kg-1 intraperitoneally. JB-9322 also dose-dependently inhibited gastric juice volume and pepsin secretion. In gastric lumen-perfused rats, intravenous injection of JB-9322 dose-dependently reduced histamine-, pentagastrin- and carbachol-stimulated gastric acid secretion. 4. JB-9322 showed antiulcer activity against aspirin and indomethacin-induced gastric lesions and was more potent than ranitidine. 5. JB-9322 effectively inhibited macroscopic gastric haemorrhagic lesions induced by ethanol. Intraperitoneal injection was effective in preventing the lesions as well as oral treatment. The oral ID50 value for these lesions was 1.33 mg kg-1. By contrast, ranitidine (50 mg kg-1) failed to reduce these lesions. In addition, the protective effect of JB-9322 was independent of prostaglandin synthesis. 6. These results indicate that JB-9322 is a new antiulcer drug that exerts a potent cytoprotective effect in addition to its gastric antisecretory activity.

Overview of the mucosal protective effects of misoprostol in man

Trimoprostil is a new synthetic prostaglandin E2 analogue that inhibits acid secretion and has mucosal protective properties. It was compared with cimetidine to assess its effectiveness in the short-term treatment of duodenal ulcer. Seven centres recruited 107 patients, who were randomized to receive either 3 mg trimoprostil daily (n = 54) or 1 g cimetidine daily (n = 53) for 4 weeks, the drugs being taken in four divided doses. Of patients completing treatment, 23 of 40 (58%) healed with trimoprostil, compared with 47 of 53 (89%) with cimetidine (p less than 0.001). Both drugs relieved daytime and nighttime pain, but cimetidine was significantly quicker. Eight patients taking trimoprostil were withdrawn because of pain, nausea, and vomiting, but none taking cimetidine; diarrhoea did not occur with trimoprostil. There were no clinically significant changes in haematology or in biochemistry studies. In conclusion, trimoprostil was not as effective as cimetidine in the treatment of duodenal ulcer.

CP-66,948 is a histamine H2-receptor antagonist with gastric antisecretory activity and mucosal protective properties. The affinity of CP-66,948 for the guinea pig atria histamine H2-receptor is 15 times greater than that of cimetidine and seven times greater than that of ranitidine. In vivo, the ED50 value for inhibition of gastric acid secretion in pylorus-ligated rats is 2 mg/kg intraduodenally, and in histamine or pentagastrin-stimulated Heidenhain pouch dogs the antisecretory ED50 values are 0.3 mg/kg per os and 1.0 mg/kg per os, respectively. CP-66,948 also inhibits ethanol-induced gastric hemorrhagic lesions in rats following either oral or systemic administration (ED50 values of 12 mg/kg per os and 6 mg/kg subcutaneously). In addition, the mucosal protective activity is independent of prostaglandin synthesis. CP-66,948 inhibits gastric acid secretion in man, and its mucosal protective activity may provide additional benefits in peptic ulcer therapy.

The gastric antisecretory effects of three different doses of enisoprost, a new synthetic PGE1 analogue, were compared with placebo and misoprostol in 20 healthy male volunteers. Enisoprost 100, 200 and 400 micrograms all significantly (P less than 0.0001; ANOVA) suppressed histamine-stimulated acid and pepsin output when compared with placebo or misoprostol 200 micrograms. Misoprostol produced a significant decrease of stimulated acid output when compared with placebo (P = 0.0012). The concentration of pepsin in gastric juice was significantly (P less than 0.0001) decreased by enisoprost at the commencement of histamine stimulation. This effect was short-lived, and was maximal with enisoprost 400 micrograms. There was a significant dose-response relationship for enisoprost for inhibition of stimulated acid output (P = 0.0065). Enisoprost was well tolerated, and no consistent drug-related adverse effects were detected. The profile of antisecretory effect of enisoprost, producing marked suppression of both acid and pepsin secretion independently, is unusual. This combination of activity along with any mucosal protective properties might be particularly effective in the treatment of peptic ulcer disease.

Sa1427 Lafutidine, an H2 Receptor Antagonist with Mucosal Protective Properties, Attenuates 5-Fluorouracil-induced Intestinal Mucositis in Mice by Reducing Secondary Inflammation via Activation of Sensory Afferent Neurons

Background: The nephroprotective effects of herbal extracts have increasingly gained attention due to the rise in kidney diseases and the limitations of conventional treatments. This study explores the renal protective properties of a polyherbal extract formulation composed of Ziziphus spina-christi, Trigonella foenum-graecum and Nigella sativa (ZTN) in Wistar albino rats. Methods: Three groups of adult male and female Wistar albino rats consisting of the control group that received only water, another group was administered with 100 mg/kg of the polyherbal (ZTN herbal) formulation, while the last group was administered with 250 mg/kg of the ZTN polyherbal extract formulation. The treatments administered for 14 days and thereafter Kidney weight, uric acid, Blood Urea Nitrogen (BUN) and creatinine levels were assessed to assess renal function. Histological analysis was conducted for tissue integrity evaluation. Result: The polyherbal extract formulation of ZTN induced a dose-dependent increase in kidney weight in both male and female rats. Uric acid levels decreased significantly in a dose-dependent manner, with the most substantial reduction at 250 mg/kg. BUN levels also showed a notable decrease, particularly at the higher dose, indicating enhanced renal function. Creatinine levels exhibited a significant dose-dependent reduction, with female rats showing higher baseline levels but greater overall reductions. Histological analysis revealed mild histological changes at 100 mg/mL and severe changes at 250 mg/mL. Histological changes include glomerular hypertrophy and tubular necrosis, more pronounced at higher doses. The polyherbal extract formulation of ZTN demonstrated significant nephroprotective effects, reducing uric acid, BUN and creatinine levels in a dose-dependent manner. Interestingly, an observation of gender-based differences occur in female rats showing higher baseline levels but greater reductions with treatment. These findings support the potential therapeutic use of polyherbal extracts in managing renal health, emphasizing the importance of dose regulation to avoid adverse effects.

Polyherbal formulations are increasingly investigated for their synergistic antioxidant potential against oxidative stress–related disorders. This study evaluated a polyherbal ethanol extract derived from Abelmoschus esculentus pods and Telfairia occidentalis leaves (AETO-PHF) through integrated in vitro and in silico approaches. GC–MS analysis identified 37 compounds, with dodecanoic acid (16.24%) and 9-octadecenoic acid (Z)-,2,3-dihydroxypropyl ester (16.41%) as predominant constituents. Antioxidant assays revealed potent dose-dependent radical scavenging, with IC₅₀ values of 61.39 ± 0.17 µg/mL (DPPH), 11.15 ± 0.15 µg/mL (H₂O₂ scavenging), 61.75 ± 0.00 µg/mL (FRAP), and 38.97 ± 2.66 µg/mL (NO inhibition). These results were statistically comparable (p > 0.05) to ascorbic acid (61.38 ± 0.58, DPPH; 61.71 ± 0.20, FRAP; and 38.94 ± 0.00, NO inhibition µg/mL) and gallic acid (11.30 ± 0.84 µg/mL, H₂O₂ scavenging). Molecular docking against cytochrome c peroxidase showed strong interactions of dodecanoic acid (− 5.7 kcal/mol) and 9-octadecenoic acid ester (− 6.2 kcal/mol), both surpassing the binding affinity of the reference antioxidant ascorbic acid (− 5.5 kcal/mol). Molecular dynamics simulations confirmed stable protein–ligand complexes with favorable RMSD, RMSF, and hydrogen-bond interaction profiles. These findings validate the traditional use of A. esculentus and T. occidentalis, demonstrate synergistic antioxidant efficacy of their polyherbal blend, and provide molecular-level insights into their mechanism of action. AETO-PHF represents a promising candidate for nutraceutical and therapeutic applications against oxidative stress-related diseases, meriting further in vivo and clinical studies.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-28672-5.