Classical Ayurvedic Formulation Research Articles

Cosmeceutical assets of ancient and contemporary ayurvedic astuteness

Cosmeceuticals are topical pharmaceutical hybrids intended to enhance the beauty that provides additional healthrelated function. Skin acts as a natural barrier between internal and external environments thus play an important role in vital biological functions such as protection against mechanical/chemical damages, microorganisms, and ultraviolet damage. All most all manuscripts of ayurvedic literature are full of cosmeceutical product, but these are spread throughout manuscripts along with their rationality. In the current scenario, different companies are manufacturing ayurvedic cosmeceutical product by taking some of the ingredients of different cosmeceutical formulation mentioned in ayurvedic texts. The basic approach of these two different groups of the formulation is different from each other, but their ultimate goal is to enhance the personal appearance of a person. It is observed that classical ayurvedic cosmeceutical formulation may act through improvement of the general condition of the person as well as the direct impact of skin and hair, whereas contemporary cosmeceuticals product enhances beauty mainly by the direct impact of skin and hair than general condition of the person.

Experimental and histopathological observation scoring methods for evaluation of wound healing properties of Jatyadi Ghrita.

Introduction:Jatyadi ghrita is a classical Ayurvedic formulation indicated in the treatment of various types of ulcers.Aim:The study was designed to explore the wound healing properties of Jatyadi Ghrita in diabetes - induced rats.Materials and Methods:In the present study, diabetes mellitus was induced to 6 to 8-week-old male Wistar rats by injecting streptozotocin cut 65 mg/kg body weight intravenously by 15 min prior to the administration of Nicotinamide at 230 mg/kg body weight intraperitoneally. Animals having diabetes were used for grouping namely, diabetic control (DC), Ghrita control (GC), positive control (PC), i.e., mupirocin HCl, Jatyadi Ghrita treatment and one group of non-DC. Full-thickness excision wound was created and diameter was recorded. Daily clinical observations were recorded. A wound scoring method was developed. Wound diameter and score were recorded on days 1, 2, 3, 5, 7, 9, 12, 14 and 15. Photographs were taken at the same time interval points. Body weight and feed consumption were recorded weekly. Animals were sacrificed at regular intervals to collect the wound area tissue for histopathology analysis. Obtained data was analyzed statistically.Results and Observation:It was observed that there was no significant difference in diameter and percent change in wound healing as compared to any control. However, clinical score and histopathological changes in Jatyadi Ghrita group were improved from the second day of the study as compared to control.Conclusion:This indicates that the drug has similar wound healing activity as compared to the modern drug mupirocin HCl.

Evaluation of pharmacological and toxicological studies of an ayurvedic medicine Rasaraj Ras on biological system of the male Sprague-Dawley rats

Background: In this study, the pharmacological and toxicological effects along with possible side effects of the classical ayurvedic formulation Rasaraj Ras (RR) which is used as a traditional medicine in the treatment of hemiplegia in the rural population were evaluated. Methods: During this study, various experiments on body growth rate, organ-body weight ratio and tissue hydration indices were performed to evaluate its efficacy and toxicity. To find out the toxicological characteristic of RR, it was administered chronically to the male Sprague-Dawley rats at a dose of 40 mg/kg. After 28 days chronic administration of the RR preparation the following toxicological changes were noted. Results: All throughout the experimental period the RR treated animals were always maintaining negligible changes in body weight, but all throughout the experimental period no statistically significant increase or decrease was noted. There is a statistically significant decrease in the relative percent weight of the male rat heart. There is a statistically highly significant decrease in the absolute weight of the male rat liver. There is a statistically highly significant decrease in the relative percent weight of the male rat liver. There is a statistically significant decrease in the relative percent weight of the male rat kidney. In the tissue hydration index determination, no significant changes were noted in case of any organ. Conclusions: As RR decreases abnormally weight of several organs in body of treated rats, so it should not be administered chronically at a higher dose.

An experimental study to evaluate the antiosteoporotic effect of Panchatikta Ghrita in a steroid-induced osteoporosis rat model

Objective:The study was conducted to develop the glucocorticoid-induced osteoporosis (GIO) model in Sprague-Dawley weanling rats using different doses of methylprednisolone (MP) and evaluate the antiosteoporotic effect of a classical ayurvedic formulation, Panchatikta Ghrita (PG), in this model.Materials and Methods:Institutional Animal Ethics Committee approval was obtained. Development of model was done by subcutaneous injection of 2 doses of MP (14 and 28 mg/kg/week) for 4 weeks in 21-day old weanlings. Following confirmation of the dose of MP that induced osteoporosis, the antiosteoporotic effect of PG was tested in this model in comparison to a known antiosteoporotic agent, alendronate. Both alendronate (2.9 mg/kg/day) and PG (1.35 g/kg/day) were administered orally 2 weeks after MP - 14 mg/kg/week injection and continued for 4 weeks. Serum and urine calcium and inorganic phosphate were analyzed at weekly intervals. Animals were sacrificed after 6 weeks, and femur bones were processed to measure bone hardness and elasticity and for histological studies.Results:Rats treated with MP - 14 mg/kg/week showed optimum osteoporotic effect with no mortality as compared to MP - 28 mg/kg/week; hence, this dose of MP was used further for the efficacy study. Osteoporotic rats treated with PG 1.35 g/kg showed increase in serum calcium and inorganic phosphate levels, whereas urine calcium and phosphate levels were significantly reduced. A significant decrease in a number of osteoclasts, whereas an increase in bone hardness and elasticity was observed as compared to diseased group demonstrating antiosteoporotic effect of PG.Conclusion:PG has an antiosteoporotic effect in GIO rat model.

Preliminary Screening of a Classical Ayurvedic Formulation for Anticonvulsant Activity.

Background:Epilepsy is a serious and complex central nervous system disorder associated with recurrent episodes of convulsive seizures due to the imbalance between excitatory (glutamatergic) and inhibitory (GABAergic) neurotransmitters level in the brain. The available treatments are neither competent to control the seizures nor prevent progress of disease. Since ages, Herbal medicines have remained important sources of medicines in many parts of world which is evidenced through their uses in traditional systems of medicine i.e. Ayurveda, Siddha, Unani, Homeopathy and Chinese etc.Aim:A polyherbal formulation (containing Terminalia chebula Retz., Asparagus racemosus Willd., Embelia ribes Burm. F, Acorus calamus L., Tinospora cordifolia (Willd.) Miers, Convolvulus pluricaulis Choisy, Saussurea lappa C.B.Clarke, Achyranthes aspera L.) is mentioned in Ayurvedic classics Bhaiṣajya Ratnāvali. The aim of the study was to evaluate the anticonvulsant activity of the formulation in Maximum electroshock and Pentylenetetrazole induced convulsions in rats.Materials and Methods:In the present study, a polyherbal formulation was developed as directed by classical text and evaluated for the anticonvulsant activity using Maximal Electroshock Shock (MES) and Pentylenetetrazole (PTZ) induced convulsions in rats. Statistical comparison was done by one way ANOVA followed by the Tukey's multiple comparison test.Results:The obtained results showed that the PHF had a protective role on epilepsy. Treatment with PHF significantly improves antioxidant enzymes activities of superoxide dismutase (SOD) and glutathione (GSH) levels significantly as compared to controls. PHF also significantly decreased malonaldialdehyde (MDA) levels in the brain. Moreover, it also attenuated the PTZ-induced increase in the activity of GABA-T in the rat brain.Conclusion:These findings suggest that PHF might have possible efficacy in the treatment of epilepsy.

Toxicological Studies of the Ayurvedic Medicine “Naradiya Laksmivilasa Rasa” Used in Sinusitis

In this study, the toxicological effects along with possible side effects of the classical Ayurvedic formulation Naradiya Laksmivilasa Rasa (NMB), which is used as a traditional medicine in the treatment of sinusitis in the rural population, were evaluated. During this study, various experiments on body growth rate, organ-body weight ratio, and tissue hydration indices were performed to evaluate its efficacy and toxicity. To find out the toxicological characteristics of NMB, it was administered chronically to male Sprague-Dawley rats at a dose of 100 mg/kg. After 32 days of chronic administration of the NMB preparation, the following toxicological changes were noted. Throughout the experimental period, the NMBtreated animals showed negligible [1.04% loss (p 5 0.914) to 3.18% gain (p 5 0.753)] changes in body weight and no statistically significant increase or decrease was noted. The study involving comparison of the relative weight of the major organs of the rats revealed some significant results. There was a statistically highly significant increase in the relative percentage weight of the male rat kidney, along with an increase in the absolute weight of the male rat spleen and an increase in the relative percentage weight of the male rat spleen. In the tissue hydration index experiment, there was a statistically significant increase in the water content of the male rat liver. As NMB increases abnormally, the weight of several organs increases in the body of treated rats, so it should not be administered chronically at a higher dose.

PHARMACEUTICAL STANDARDIZATION OF APAMARGA KSHARA YOGA

Objective: Apamarga Kshara Yoga (APK), a classical Ayurvedic formulation containing Apamarga Kshara (alkaline substance of Achyranthes aspera Linn.), Manahshila (Arsenic Disulphide) and Gomutra (cow’s urine) is effective in cases of Shvitra (vitiligo). As the formulations has short shelf life, inconvenient to patients and involve longer duration of pharmaceutical preparation; it is planned to convert into more convenient form. Design: Raw materials were authenticated and APK was prepared. Further, organoleptic and physico-chemical properties were evaluated. Results: An average 909.8 gm (90.98 %) yield after Taila Paka and 1379.6 gm ointment was observed in the process. Conclusions: As observed in present study, addition of wax to attain ointment consistency is mandate. Residue as Apamarga Kshara and Manahshila incorporated in preparation of ointment yielded more without losing its consistency.

Effect of Shatpuspadya Churna - a classical Ayurvedic formulation on formaldehyde and Complete Freund’s adjuvant (CFA) induced arthritis in rats

Shatpuspadya Churna has been traditionally used in the Ayurvedic system of medicine of India to treat rheumatoid arthritis (RA). We evaluated anti-arthritic activity of ethanol extract of Shatpuspadya Churna (SCE). Arthritis was induced in male rats of wistar strain by administration of formaldehyde (2%v/v) or Complete Freund’s Adjuvant (CFA) into sub-plantar surface of the left hind paw. SCE (135,270 and 540 mg/kg) was given by oral route. Joint swelling was measured in formaldehyde induced arthritis. In case of CFA induced arthritis, paw volume, body weight, haematological and histological parameters were measured. In CFA induced arthritic rats, significant (p < 001) reductions in haemoglobin (28.4 %), packed cell volume (30.5 %), HDL(45 %), and significant(p < 001) increases in WBC (28.0 %), platelet count (40.7 %), ESR (52.5 %), rheumatoid factor (274 %), total cholesterol(49.5 %), triglycerides(34.8 %), LDL (106.8 %) and VLDL (35 %) were observed. Treatment with SCE significantly (p < 0.001) reversed these changes in a dose dependent manner. Treatment with 540 mg/kg SCE significantly (p < 0.001) reversed/prevented the decline in Hb, PCV and HDL to only 7.9 %, 19.9 % and 9.8 % respectively. Similarly, the rise in values of various blood parameters was comparatively less in SCE treated arthritic group (540 mg/kg) than that observed in untreated arthritic rats. We noted only a marginal rise in the values of WBC (18.2 %), platelet count (26.5 %), ESR (40.15 %), RF (60.6 %), TC (25.4 %), TG (9.3 %), LDL (49.2 %) and VLDL (9.2 %). Histopathological studies of the ankles revealed significant improvements. The study clearly establishes the usefulness of Shatpuspadya Churna in the treatment of arthritis, lipid imbalance and anaemia associated with it.

Anti-arthritic activity of a classical Ayurvedic formulation Vatari Guggulu in rats

In India, Vatari Guggulu has been traditionally used in the Ayurvedic system of medicine to treat rheumatoid arthritis (RA). The current study was undertaken to evaluate anti-arthritic activity of alcoholic extract of Vatari Guggulu in rats. Arthritis was induced by administration of formaldehyde (2%v/v) or Complete Freund's Adjuvant (CFA) into the sub-plantar surface of left hind paw of the animals. The extract was administered to the rats by oral gavages in different doses. Joint swelling was measured in formaldehyde induced arthritis. Various physical, biochemical and histopathological parameters were determined in CFA induced arthritis. Vatari Guggulu extract (VGE) produced significant (P < 0.05) inhibition of joint swelling in both formaldehyde and CFA induced arthritis. The treatment also brought to normalcy the increased white blood cell (WBC) count, rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), cholesterol, triglycerides and LDL with an enhancement of haemoglobin (Hb) levels and red blood cell (RBC) count. These effects were found to be dose dependent. These effects were comparable with standard drug indomethacin. Histo-pathological studies of the ankles of VGE treated animals exhibited significant improvements. VGE did not show any toxic symptoms even at a dose of 2000 mg/kg in acute toxicity studies on rats. Thus, Vatari Guggulu, a classical Ayurvedic formulation of the Indian System of Medicine, exhibited significant anti-arthritic activity in formaldehyde and CFA induced arthritis in rats. This study corroborates the claims of Ayurveda on Vatari Guggulu.

Experimental evaluation of analgesic, anti-inflammatory and anti-platelet potential of Dashamoola.

Background:Dashamoola, in the form of arishta and kwath, is a commonly used classical Ayurvedic multi-ingredient formulation for management of pain, arthritis and inflammatory disorders.Objective:To study analgesic, anti-inflammatory and anti-platelet activity of Dashamoola and its combination with aspirin.Materials and Methods:Wistar albino rats (180-200 g) and Swiss albino mice (20-25 g) of either sex were divided randomly into five groups: Distilled water, aspirin (500mg/kg in rats; 722.2 mg/kg in mice), Dashamoolarishta (1.8 mL/kg in rats; 2.5 mL/kg in mice) and Dashamoolarishta with aspirin. Anti-inflammatory activity was measured by change in paw volume in carrageenan-induced inflammation, protein content in model of peritonitis and granuloma weight in cotton pellet granuloma. Analgesic effect was evaluated by counting number of writhes in writhing model. Maximum platelet aggregation and percentage inhibition of ADP and collagen-induced platelet aggregation were estimated in vitro. Statistical analysis was done using one way ANOVA (post hoc Tukey's test) and P < 0.05 was considered significant.Results:Dashamoolarishta and its combination with aspirin showed significantly (P < 0.01) less number of writhes. It showed significant (P < 0.001) anti-inflammatory activity by paw edema reduction in rats, decrease in proteins in peritoneal fluid (P < 0.001) and decrease in granuloma weight (P < 0.05) as compared to respective vehicle control groups. Dashamoola kwath alone and in combination with aspirin inhibited maximum platelet aggregation and percent inhibition of platelets as compared to vehicle (P < 0.001).Conclusion:Dashamoola formulation alone and its combination with aspirin showed comparable anti-inflammatory, analgesic and anti-platelet effects to aspirin.

Clinical efficacy of Vyaghriharitaki Avaleha in the management of chronic bronchitis.

Introduction:Vyaghriharitaki Avaleha (VHA), a polyherbal classical Ayurvedic formulation has been used in Kasa (cough), Swasa (asthma), Rajayakshma (tuberculosis) etc., conditions.Aim:To evaluate the clinical efficacy of VHA in the management of chronic bronchitis.Materials and Methods:Patients of chronic bronchitis were given 10 g of VHA twice a day with lukewarm water before meals for 12 weeks. Improvement in clinical symptoms of chronic bronchitis as the primary outcome measures and St. George's Respiratory Questionnaire scores as secondary outcome measure was studied.Results:Out of 66 enrolled patients, 61 completed the treatment schedule. The results show that VHA provides statistically significant improvement with P < 0.001 in both primary and secondary outcome measures.Conclusion:VHA can be considered as a safe and effective formulation in the management of chronic bronchitis.

Design, Development and Rationalization of Sarpagandha Ghanvati.

Sarpagandha ghanvati is a classical Ayurvedic formulation widely prescribed for anxiety and insomnia. It contains Sarpagandha (roots of Rauwolfia serpentina L. (Benth.) Ex Kurz; Family: Apocyanaceae), Khurasani ajowan (Hyocyamus niger L.; Family: Solanaceae) seeds, Jatamansi (Nardostachys jatamansi DC. Family: Valerianaceae) roots and Pipplamul (root of Piper longum L.; Family: Piperaceae). The objective of this study was to make a comparative evaluation of Ghanvatis and tablets of this formulation. Two tablet formulations were prepared; one incorporating only powders of all ingredients; the other with ethanol extracts of the first three ingredients and powder of Piper longum root. Similarly, two types of Sarpagandha ghanvati pills were prepared; one as per Ayurvedic Formulary of India; the other with ethanol extracts of the first three ingredients and powder of Piper longum root. Alcohol extracted 0.22% w/w of total alkaloids as against 0.061% w/w extracted by water. Tablets prepared with powders of all the ingredients had friability more than 3.0% where as those prepared with ethanol extract had very low friability. Ghanvatis, prepared as per the Ayurvedic formulary, did not show reserpine although other alkaloids were present. They showed less content uniformity and lower drug release. Ethanol extracted reserpine along with other alkaloids. Ghanvatis made with the alcoholic extracts exhibited better content uniformity and drug release than the traditional formulation. Tablets prepared with powders or extracts of the ingredients exhibited good content uniformity but the release of alkaloids from the tablets of powders was only 80%. Tablets of the extracts had good content uniformity with 90% release of the total alkaloids. Tablets prepared with alcoholic extracts using 1% polyvinylpyrrolidone as binder and 5% dried starch powder as disintegrating agent confirmed to all the requirements. Thus, the study shows tablets made with the extracts are superior to Ghanvatis and powder tablets.

QUANTITATIVE DETERMINATION OF THREE CONSTITUENTS OF RASAYANA CHURNA (A CLASSICAL AYURVEDIC FORMULATION) BY A REVERSED PHASE HPLC

India has long tradition of using Ayurvedic medicines as therapeutic agent. In comparison to rapid growth of herbal market, export rate of traditional Indian medicine is quite insignificant. It might be due to lack of proper quality control measures of raw materials and formulation. So, it is essential to ensure quality, safety and efficacy of raw materials and f ormu l ation. Rasayana Churna is a classical Ayurvedic formulation, which comprises dried powders of three well known rejuvenating drugs viz. dried stem of Guduchi ( Tinospora cordifolia Miers.), dried fruit of Gokshur ( Tribulus terrestris Linn.) and dried pe ricarp of Amalaki ( Emblica officinalis Gaertn.) in equal proportion. It is used in Ayurveda as Rasayana to enhance general body re sistance, promote longevity, as anti - stress and adaptogen. T herapeutic activity of Rasayana Churna may be attributed to cordif olioside A, diosgenin and gallic acid. Cordifolioside A is an active constituent of Guduchi ( Tinospora cordifolia Miers.). Diosgenin is steroidal saponin present in Gokshur ( Tribulus terrestris Linn.) and gallic acid is a phenolic compound present in Amala ki ( Emblica officinalis Gaertn.). Several research work have reported the quantitative estimation of active constituent in individual drug but no established method was found for quantit ative analysis of Rasayana Churna . This is the first attempt ever rega rding quantitative estimation of Rasayana Churna by RP - HPLC. Various validated methods were used for estimation of cordifolioside A, diosgenin and gallic acid. These RP - HPLC methods can be used for routine quality control of raw materials and

PUNARNAVA MANDUR: TOXICITY STUDY OF CLASSICAL AYURVEDIC FORMULATION IN WISTAR RATS

Punarnava Mandur is an iron containing classical Ayurvedic formulation which was studied for repeated dose oral toxicity study in Wistar rats for 90 days. Total 48 Wistar rats (24 male and 24 female) were selected based on the body weight and were randomly distributed into four groups followed by administration of Punarnava Mandur at the dose of 0, 90, 450, 900 mg/kg body weight for 90 consecutive days. Body weight, Weekly Feed and Water consumption, Clinical Chemistry, Hematology, Differential leukocyte Count, Reticulocyte count and Organ weights were recorded and analyzed statistically. At termination, rats were sacrificed, examined for gross pathological changes, organs were collected, weighed and processed for histopathological evaluation. There was no effect on body weights and feed consumption, no abnormal findings in the histopathological evaluation of high dose group animals but there was significant increase in weight of liver in females of high dose group as compared to control. Hence, the dose level 450 mg/kg of Punarnava Mandur was found as NOAEL (No Observable Adverse Effect Level). However, the NOEL (No Observed Effect Level) could not be established. It was suggested to carry out a toxicity study at possible higher doses so as to establish target organ of toxicity.

PA02.12. Toxicity studies on shankha vati

Purpose:Shankha Vati, a classical Ayurvedic formulation that is widely used by the practitioners of Ayurveda, contains a heavy metal i.e. Parada (mercury) and a poisonous herbal drug i.e. Vatsanabha (Aconitum chasmantum Staff.ex.Holmes) along with other ingredients. There are certain apprehensions on this formulation, mainly due to the presence of its ingredient Vatsanabha. Shankha Vati (Bhaishajya Ratnavali 10/186 – 187, also in Ayurvedic Formulary of India Part1), is one of the formulations in which Vatsanabha is used without its usual coingredient and antidote Tankana (borax). With an hypothesis that classical combination of Shankha Vati is not completely safe and there could be some adverse effects due to the presence of poisonous ingredient Vatsanabha in it, and any possible adverse effects of Shankha Vati due to the presence of Vatsanabha will be nullified by the addition of Tankana to the formulation, another modified formulation with the addition of purified Tankana equal to the quantity of Vatsanabha in the classical Shankha Vati, was also taken up for the study.Method:Acute toxicity study as per OECD Guideline 425, and Chronic toxicity study (90 days with the recovery period of 30 days) as per AYUSH170 guideline (Guidelines for preclinical safety evaluation for Ayurveda, Siddha and Unani drugs and other traditional medicines in India) were carried out. Biochemical and haematological investigations were carried out on 30th, 60th, 90th and 120th days. Histopathological study of acute chronic toxicity study were carried out.Result:Histopathological study of both acute and chronic toxicity study were also carried out. There were no toxicity related changes found in the animals treated classical Shankha Vati as well as modified Shankha Vati in both acute and chronic toxicity study. Similarly biochemical and haematological investigations also did not toxicity related changes.Conclusion:Classical formulation of Shankha Vati is completely safe and there is no need to modify the composition to make it safer.

Effect of Rasayana Ghana tablet (An Ayurvedic formulation) on improving quality of life of stressed individuals

Stress is the most hazardous factor which can adversely affect the quality of life. Ayurvedic Rasayana formulations can be used to relieve stress and there forth improve and maintain healthy quality of life. In the present study, the effect of a classical ayurvedic Rasayana formulation comprising three herbs Gokshura (Tribulus terrestris Linn. ) , Aamalaki (Emblica officinalis Gaertn. ) and Guduchi (Tinospora cordifolia Miers. ) is assessed on health, wellness and quality of life questionnaire in stress prone patients. Total 108 patients complaining of stress and decreased quality of life were randomly divided into two groups of Rasayana Ghana tablets (RGT) and Placebo group (PG). The drug was given in a dose of two tablets of 500mg twice a day with ghee and honey in unequal proportion. Result was assessed on the parameters given in quality of life questionnaire. The data was then analyzed by applying Wilcoxon non parametric test in each group and chi-square test to compare efficacy in both groups. RGT seems to be effective in decreasing the physical complaints and improving mental/emotional state, adaptation to stress, life enjoyment and overall quality of life to a significant level in each sub scale.

Hepatoprotective activity of punarnavashtak kwath, an Ayurvedic formulation, against CCl4-induced hepatotoxicity in rats and on the HepG2 cell line

Objective: Punarnavashtak kwath (PNK) is a classical Ayurvedic formulation, mentioned in Ayurvedic literature Bhaishajya Ratnavali, for hepatic disorders and asthma. This study investigated the hepatoprotective activity of PNK to validate the traditional use of this formulation.Materials and methods: PNK was prepared in the laboratory according to the method given in Ayurvedic literature. Phytochemical screening was performed to determine the presence of phytoconstituents. Hepatoprotective activity was evaluated against CCl4-induced hepatotoxicity in rats and by its effect on the HepG2 cell line.Results: Preliminary phytochemical screening revealed the presence of alkaloids, tannins, flavonoids, saponins, and a bitter principle in PNK. Administration of PNK produced significant hepatoprotective effect as demonstrated by decreased levels of serum liver marker enzymes such as aspartate transaminase, serum alanine transaminase, serum alkaline phosphatase, and serum bilirubin and an increase in protein level. Thiopentone-induced sleeping time was also decreased in the PNK-treated animals compared with the CCl4-treated group. It also showed antioxidant activity by increase in activity of glutathione, superoxide dismutase, and catalase and by a decrease in thiobarbituric acid reactive substance level compared with the CCl4-treated group. Results of a histopathological study also support the hepatoprotective activity of PNK. Investigation carried out on the HepG2 cell line depicted significant increase in viability of cells exposed to PNK as compared with CCl4-treated cells.Discussion and Conclusion: It can be concluded that PNK protects hepatocytes from CCl4-induced liver damages due to its antioxidant effect on hepatocytes. An in vitro study on HepG2 cell lines also supports its protective effect.

In vivo and In vitro Antioxidant and Hepatoprotective effects of Classical ayurvedic formulation Punarnavashtak kwath against Ethanol induced hepatotoxicity

Punarnavashtak kwath (PN) a classical Ayurvedic formulation reported in “Bhaishyajyaratnavali” consisting of eight medicinal plants was evaluated for its in vitro and in vivo antioxidant activity and hepatoprotective effect. In vitro antioxidant activity of PN kwath was investigated by DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity, superoxide anion and nitric oxide radical scavenging activity and reducing power assay. In vitro antioxidant activity was found to be dose dependent. Hepatoprotective and in vivo antioxidant effect was evaluated by ethanol (3.7 g/kg, p.o for 45 days in all group except control) induced hepatic damage in rats. Pretreatment with PN kwath 100 mg/kg p.o for 45 days) significantly prevented physical (increased liver wt), biochemical (serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), bilirubin and total protein level), histological (damage to hepatocytes) and functional changes (thiopentone induced sleeping time) induced by ethanol in liver. Further PN kwath showed antioxidant activity by increasing activity of GSH, SOD and CAT and by decreasing the level of thiobarbituric acid reactive substance (TBARS). The results were compared to that of reference standard silymarin (50 mg/kg p.o for 45 days). The findings suggest that PN kwath protects the liver cell from ethanol induced liver damages due to its antioxidative effect on hepatocytes.

Estimation of biomarkers berberine and gallic acid in polyherbal formulation punarnavashtak kwath and its clinical study for hepatoprotective potential.

Punarnavashtak (PN) kwath is a classical Ayurvedic formulation mentioned in Ayurvedic literature “Bhaishyajyaratnavali” for hepatic disorders and asthma. Standardization and clinical trial to support its efficacy are lacking. So, in the present study, standardization of PN kwath was done by using biomarkers, gallic acid and berberine, and its hepatoprotective activity was evaluated by clinical study to rationalise the traditional use of this formulation. PN kwath was standardized by HPTLC (High performance thin layer chromatography) using gallic acid and berberine as biomarkers and was subjected to clinical study. For clinical study patients attending outpatient clinics, with an evidence of liver disease were included in the study. During the study period, patients who fulfilled inclusion criteria were randomly assigned. The recommended dose was 20 ml kwath daily for 8 weeks. All the patients underwent clinical examination and laboratory investigations for liver functions tests before the commencement of therapy. Thereafter, clinical assessments were done after 8 weeks of treatment. The results showed significant changes in liver functions tests [serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), total bilirubin]. There was no report of adverse effects attributable to this formulation. Our results suggest that PN kwath showed significant hepatoprotective activity. Berberine and gallic acid were found to be 0.08 and 4.9%, respectively. Our results suggest that PN kwath showed significant hepatoprotective activity due to presence of various phytoconstituents and support its traditional uses in liver disorder. Key words: Berberine, clinical study, gallic acid, hepatoprotective, punarnavashtak kwath

Validation of Preparation Process and Development of Fingerprints for Ayurvedic Formulation Arjunaristha by High Performance Thin Layer Chromatographic Estimation of Gallic Acid

Arjunaristha (AJR) is a classical Ayurvedic formulation used as general tonic for heart disease, lung diseases, loss of strength, immunomodulator and in azoospermia. It is observed that the consistency and taste of the formulation varies from manufacturer to manufacturer. Hence, it is need of the hour to develop standardized preparation process and validation, with the advent of new analytical tool and sophisticated instrumental technology. Three batches of AJR were prepared in the laboratory according to method given in Ayurvedic formulary of India and three marketed samples procured from local pharmacy. Process validation was developed with the help of physiochemical and chromatographic studies at every step of the preparation process. AJR contains Terminalia, Arjuna and Woodfordia flaburanda, containing gallic acid as main active constituents. The high performance thin layer chromatographic method has been developed for the fingerprinting of AJR. The result of different stage samples (laboratory) found in close proximity, while finished formulation and marketed samples showed little deviation. The methods used for validation of preparation process and fingerprinting were found to be precise and reproducible and can be used for the routine process validation and fingerprinting of AJR.