Standardization and validation of phytometabolites by UHPLC and high-performance thin layer chromatography for rapid quality assessment of ancient ayurvedic medicine, Mahayograj Guggul.

Mahayograj Guggul is an ancient ayurvedic medicine, prescribed for various joint disorders like arthritis, gout, and rheumatism. The present research was envisaged to develop a simple, sensitive, and comprehensive analytical method for standardization of Mahayograj Guggul. The analysis was conducted for gallic acid, protocatechuic acid, vanillic acid, cinnamic acid, piperine, guggulsterone-E, and guggulsterone-Z by high-performance thin-layer chromatography, and additionally ferulic acid, ellagic acid, and picroside I by ultra high-performance liquid chromatography. These developed methods were validated as per international guidelines, and were found linear (r2  > 0.99), sensitive, precise (relative standard deviation < 5%), and accurate with recovery values (85-105%). The limit of detection and quantification were in the range of 0.11-23.6 and 0.33-71.51 µg/g. Gas chromatography tandem mass spectrometry was used to develop Mahayograj Guggul fingerprinting profile and to identify mid-polar or nonpolar compounds. Proximate analysis was used to ascertain the functional groups present in Mahayograj Guggul. Ultra high-performance liquid chromatography and gas chromatography tandem mass spectrometry were further employed to authenticate quality reproducibility in the active ingredients of Mahayograj Guggul in six commercial batches. Taken together, these analytical methods provide a scientific basis and reference for quality control evaluation of Mahayograj Guggul and similar traditional broad-spectrum formulations.

Development of a reliable Dosha self-assessment questionnaire.

INTRODUCTION: The Ayurvedic theory of Tri-Dosha offers a unique framework for understanding human differences in health and disease. The Doshas could be readily applied in clinical studies to control for such differences the same way sex, age, height and weight are routinely used except that clinical evaluation is required and methods vary. No open-source questionnaire has been shown to be reliable for self-assessment. Therefore, we undertook this study to develop a reliable Dosha self-assessment questionnaire with characteristics suitable for use in clinical research. METHODS: A review of published questionnaires and translations of classic Ayurvedic texts along with an analysis of item-total correlations and ANOVAs by Dosha for 735 responses to a 37-item ipsative questionnaire from an Ayurveda products company served as the basis for generating 108 Dosha-specific test items featuring a 7-point Likert scale. We used item-total and inter-item correlations from a convenience sample of 176 individuals to identify the best performing items. We then solicited repeat responses to the resultant 39-item questionnaire (13 for each Dosha) from these and other website visitors and calculated test-retest reliability as the intraclass correlation coefficient for absolute agreement (ICC 2,1) for each Dosha. RESULTS: The final questionnaire gives normally distributed results. Among 76 self-selected adults, the test-retest reliabilities for Vata, Pitta and Kapha Dosha scores are all above 0.8. CONCLUSIONS: This study demonstrates that it is possible to quantify the three Doshas with adequate reliability for most clinical studies.

Assessment of the Cytotoxic Activity of Triphala: A Semisolid Traditional Formulation on HepG2 Cancer Cell Line.

Cancer chemotherapies may result in resistance, and therefore, contemporary treatments including natural products may find an increasing consideration. As per Persian medicine (PM), many natural products have been used for malignant and chronic diseases. Triphala, with a combination of Terminalia chebula Retz., Terminalia bellirica Retz., Phyllanthus emblica L., and honey, is a multi-ingredient traditional formulation attributed to anticancer activities in PM. This study is aimed at evaluating the cytotoxic activity of this preparation on HepG2, the human liver cancer cell line. Hydroalcoholic extracts were prepared from the formulation and its components. Compared with the control and Cisplatin, the extracts were tested using MTT assay at different concentrations. All concentrations of the preparation, as well as Cisplatin, were effective significantly against HepG2 cells. All extract preparations at multiple concentrations were significantly effective as evidenced by MTT assay when compared to the control group. The IC50 level for Triphala extract was 77.63 ± 4.3 µg/ml. Based on the results, Triphala and its components have cytotoxic activity on the HepG2 cancer cell line and they can reduce the survival rate significantly.

Promoting health economic evaluation studies in the AYUSH system of medicine: the need of the hour.

The double burden of communicable and noncommunicable diseases is a major threat to the Indian public health system. AYUSH, an acronym for Ayurveda, Yoga and Naturopathy, Unani, Siddha, Sowa-Rigpa, and Homeopathy, represents the Indian system of medicine recognized by the Government of India. Mainstreaming of AYUSH is one of the key strategies of the Indian government for tackling increasing disease burden through initiatives such as AYUSH wellness centers, telemedicine, and quality control measures for medications in the AYUSH system of medicine. Such investment of resources in health systems may require economic evaluations. However, such evaluations are lacking in the AYUSH system, except for a few in homeopathy and yoga. In the absence of evidence from economic evaluations, researchers and decision makers are guided mostly by clinical efficacy while formulating healthcare strategies. In view of the increasing use of AYUSH across the country, economic evaluations of the AYUSH system are the need of the hour to aid healthcare decision making.

Assessment of essential and non-essential elements in selected traditional medicines from India, Ghana and China.

Traditional medicine (TM) also known as folk medicine consists of medical knowledge systems that were developed over generations in various countries before the era of modern medicine. Over the last few years, there has been a shift in universal trend from modern medicine to TM because it is believed that these medicines are non-toxic to humans, have little to no side-effects, are readily available and affordable. The purpose of this study was to investigate the essential and non-essential elemental concentrations of ayurvedic medicines (AM), Ghanaian traditional medicines (GTM) and Chinese traditional medicines (CTM). The results indicate that the mean elemental concentrations in TM varied greatly. The mean concentrations of copper and mercury in AM, GTM and CTM ranged from 4 to 45 mg/kg and 0.01 to 2 mg/kg, respectively. Multivariate statistical analyses applied to recognise inter-element associations indicated that at 0.05 significant level, there was a positive correlation between elements suggesting they may have originated from the same source. The study concludes by highlighting the need for monitoring of the elemental concentrations in commonly consumed traditional medicines regularly to detect contamination in these medicines since the occurrence of metals in these medicines above or below legally permissible limits can be harmful to consumers.

Quality Assessment of Sitopaladi Churna Using High-Performance Liquid Chromatography Coupled with Multivariate Analysis.

"Sitopaladi churna," a well-known formulation of Ayurveda, is prescribed to treat the disease like Bronchitis, Pneumonia, allergic conditions, viral infection of the respiratory tract and as a natural antioxidant. A novel method based on reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to photodiode array detector was established and validated for sitopaladi churna. Here, HPLC fingerprints data obtained for 28 samples including standard sample were then treated to chemometric analysis like principal component analysis and hierarchical clustering analysis for further analysis to evaluate the differences in market samples of sitopaladi churna. Additionally, one major marker compound, piperine was quantified and it also facilitated for relative retention time. The simulative mean spectrum was also generated. The validation results showed that the developed method was simple, precise and stable. Thus, the developed chromatographic method adjoined with multivariate analysis can be used as an efficient and practical approach for quality assessment of sitopaladi churna.

Patient Assessment and Chronic Pain Self-Management in Ethnomedicine: Seasonal and Ecosystemic Embodiment in Ayurvedic Patient-Centered Care.

Background: Ayurveda's preventive focus complements its strength with the interventionist approach of the biomedical in chronic pain self-management. Patient-centered care (PCC) using ethnomedicine promises greater patient self-management; however, few studies have examined environmental relationships and PCC in self-management of chronic pain through Ayurveda. Objective: To examine how Ayurveda's philosophical focus on whole system frameworks describes the integration of the individual and the ecological in tailoring an integrative patient-centered diagnostic and prognostic approach to chronic pain management. Methods: This qualitative case study conducted in-depth semi-structured interviews of Ayurvedic physicians from India (N = 10) and a qualitative inductive content analytic approach. Findings: The diagnostic and interpretational framework of the doshas supports the integration of the individual and the ecological through (a) the circadian and seasonal cycles relating mind-body awareness with diet, lifestyle (e.g., yoga), and breath (e.g., pranayama), and (b) biogeographical and ecosystemic regions relating the biogeographical and the ecological (e.g., desh) with the regulatory principle of pain and its physiological and anatomical perception (vata) in an approach that goes beyond treating pain etiology to a whole person PCC approach. Conclusions: The study highlights how circadian and seasonal cycles and evolutionary spatial-temporal factors of biogeographical and ecological regions are employed in patient assessment and self-management to support patient involvement. Recommendations for PCC in integrative chronic pain management include supporting patient ownership of their care through the dosha framework that relates the individual and the ecological in the patient's own life-context and supports co-creation of a collaborative plan of care using an ethnomedical framework.

Health System Development in Nepal.

The development of health system in Nepal dates back from ancient period to the modern period starting from ayurvedic medicine to today's modern allopathic treatment. With regard to the different rulers in different time period, the number of improvements and changes in the health system of Nepal has been made. Moreover, the health system is further strengthened by the involvement of people and better management of health information and drug supply. Keywords: Bir Hospital; Community health; Health; Nepal.

A simple, cost-effective and rapid method for simultaneous determination of Strychnos nux-vomica alkaloids in blood and Ayurvedic medicines based on ultrasound-assisted dispersive liquid-liquid microextraction-thin-layer chromatography-image analysis.

A simple, rapid, cost-effective and green analytical method is developed based on ultrasound-assisted dispersive liquid-liquid microextraction (US-DLLME) coupled to thin-layer chromatography (TLC)-image analysis for the simultaneous determination of two major alkaloids of Strychnos nux-vomica L i.e., strychnine and brucine. The method is composed of three steps, namely (i) US-DLLME by injecting a mixture of 100-µL chloroform (extraction solvent) and 1-mL methanol (disperser solvent) in 5 mL of aqueous sample, followed by ultrasonication and centrifugation, (ii) TLC of 20 µL of sedimented phase with methanol: ammonia (100:1.5, v/v) as the mobile phase and visualization under ultraviolet radiation (254 nm) and (iii) photography of TLC plate and quantification of spots by image analysis using freely available imageJ software (National Institute of Health, Bethesda, MD, USA). The limit of detection and limit of quantification for both alkaloids were found to be in the range of 0.12-0.15 and 0.36-0.48 µg/spot, respectively. The method was found to be linear in the range of 0.5-5 µg/spot with correlation coefficient (R2) of 0.995 and 0.997 for strychnine and brucine, respectively. The developed method was successfully applied for the determination of strychnine and brucine in Ayurvedic formulations and blood samples. The method does not require any sophisticated instrument and handling skills and can be adopted for rapid analysis of strychnine and brucine in forensic toxicological laboratories.

Physicochemical characterization of Suvarna Bhasma, its toxicity profiling in rat and behavioural assessment in zebrafish model.

ETHNOPHARMACOLOGICAL RELEVANCE: Suvarna Bhasma is a gold-based Ayurved medicine that has a wide range of therapeutic indications like tuberculosis, diabetes mellitus, rheumatoid arthritis and nervous diseases. Suvarna Bhasma is also used in Suvarnaprashana, an Ayurved advocated therapy being practised to improve immunity in children. AIM OF THE STUDY: To augment traditional understanding, here we present an evidence-based study on Suvarna Bhasma regarding its physicochemical properties, toxicity and efficacy. MATERIALS AND METHODS: Suvarna Bhasma was characterised by physicochemical characterization techniques such as scanning electron microscope (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and atomic emission spectroscopy (ICP-AES). Toxicity of Suvarna Bhasma was studied in Holtzman rats with daily oral dose from 3 mg/kg (therapeutic dose, TD) up to 30 mg/kg (10 TD) body weight for 90 days. Behavioural study, such as motor and geotactic behaviour were examined in zebrafish model to find out any sign of neurotoxicity or behavioural changes due to Suvarna Bhasma administration. RESULTS: Suvarna Bhasma has two types of gold particles, large ones (~60 µm) having irregular shapes, and nano-sized spherical particles (starting from ~10 nm), the latter coated with Fe, Si, O, P and Na. XRD study revealed that all the peaks of Suvarna Bhasma match well with pure gold (face centred cube) with crystallites size 45 ±â€¯2.8 nm. In rat studies, some change in biochemical parameters such as urea, creatinine and alanine aminotransferase (ALT) was observed mainly at the higher therapeutic dose; however, those parameters were within the normal range. There were no significant macroscopic as well as microscopic treatment-related alteration observed, in any of the organs and tissues evaluated. In zebrafish behavioural study, the motor parameters of Suvarna Bhasma treated fish showed normal behaviour analogous to the vehicle control group. Interestingly, the geotactic behaviour showed anxiolytic effects of Suvarna Bhasma as evidenced by the time spent in the upper zone, and average swimming height. The anxiolytic effects persisted for more than 30 days after withdrawing the Suvarna Bhasma treatment. CONCLUSIONS: Suvarna Bhasma contained spherical gold nanoparticles. It was nontoxic in rat model at the does tested. Suvarna Bhasma has anxiolytic effects in zebrafish behavioural model.

Assessment of Anxiety, Depression, Stress, and Associated Psychological Morbidities among Patients Receiving Ayurvedic Treatment for Different Health Issues: First Study from Sri Lanka.

BACKGROUND: Good mental condition is a vital part of health. Physical impairments would potentially have psychiatric manifestations during the course of a disease that could cause patients to experience a wide range of psychological conditions. This study was conducted to determine prevalence of anxiety, depression, stress, and psychological morbidities among the patients who received warded treatments at Gampaha Wickramarachchi Ayurveda Teaching Hospital, Sri Lanka. METHODS: A total of 148 patients admitted to the hospital were selected for the study on a random systematic basis under four systemic groups (gastrointestinal, integumentary, musculoskeletal, and nervous system) depending on the chief complaint. The presence of depressive, anxiety, and stress symptoms was assessed by the Depression Anxiety Stress Scale 21 item version (DASS 21). The General Linear Model (GLM) was used for statistical analysis. RESULTS: Over 50% of the participants in all four patient groups belonged to age group of 35 to 65 years, encompassing the fraction of population that actively contribute to the workforce in the society. Stress, anxiety, and depression values of patients belonging to different complications varied significantly, as indicated by GLM (p < 0.05). Patients diagnosed with integumentary system-related issues denoted the highest stress levels (27.7 ± 2.54), while the mean stress values among the other systemic groups were not significantly different among each other. The highest anxiety levels were indicated by patients with nervous system-related issues (18.6 ± 1.51), while the lowest anxiety levels were indicated by patients with integumentary disorders (6.0 ± 2.73). The highest depression level was identified from patients suffering from integumentary system-related disorders (31.7 ± 3.42), followed by nervous system (23.2 ± 1.78), gastrointestinal (19.5 ± 3.77), and musculoskeletal (16.8 ± 1.57) disorders. CONCLUSION: Overall, high distress levels were observed among the majority of the patients. Furthermore, integumentary issues may lead to significant psychological impacts. As most of the patients seek for Ayurveda treatments when their diseased condition becomes chronic, it is vital to focus on a biopsychosocial approach to patient assessment and patient care, in actual practice.

Market Analysis of Medicinal Plants in India.

There has been significant growth in the herbal drugs market around the world, over the last few decades due to growing awareness among people about the rising cost as well as side effects related to the use of synthetic drugs. Herbal medicine has been used in traditional medicinal systems around the world, especially India where the oldest systems of medicine namely Ayurveda, Siddha, and Unani make use of more than 90% plant-based formulations. India is rich in medicinal and herbal plants resources that provide for both the health care needs of rural India as well as the source of raw material for therapeutic agents required in the production of traditional and modern medicine. This review article presents an overview of the knowledge of Indian herbal plants based medicine in the national and international market and the trends in its production, sustainability, and promotion.

Assessment of compliance level of ICH guidelines for organic volatile impurities in common ayurvedic hepatic formulations.

Background Herbal medicines have been used in the treatment of liver diseases for a long time. In recent years, the use of herbal medicines for protection from other strong antibiotics as well as drugs that can damage the liver during their metabolism in liver and for treatment of liver diseases has increased all over the world. It is important to mention that a number of organic solvents are used at different stages of extraction/formulation development for these traditional preparations in industries/pharmacies. In addition, some of these solvents possess established carcinogenic properties and may enter the formulation as residual solvents. Hence as per ICH guidelines it is mandatory to keep the level of these solvents up to permissible limits. There has been a lot of hue and cry that ayurvedic formulations available in the market are not properly standardized for their quality due to lack of stringent regulations and standards from regulatory authorities. Therefore the aim of present work was to assess the compliance of ICH guidelines for level of organic volatile impurities in common marketed ayurvedic hepatic formulations. Methods In this study, 25 ayurvedic herbal formulations available as OTC product have been assessed for presence of residual solvents using gas chromatography with flame ionization detector. Results This study on 25 fast moving hepatic formulations in the market reflects that no residual solvents were detected in any of the formulations however if present were within prescribed permissible limits of ICH guidelines. The data was also subjected to statistical analysis (F-test and t-test at 95% confidence level). Conclusions Results indicate the safety of these hepatic formulations with respect to residual solvents. In addition presents a simple, linear, specific, accurate, precise and rugged gas chromatographic method for estimation of residual solvents.

Regulatory Aspects of Traditional Indian Medicines (TIM) in India and in International Purview.

Background: Regulatory affairs play a crucial role in the pharmaceutical industry and are incorporated in all stages of drug development. Objective: Approval criteria practices were developed as a resolution of the government's desire to create policies to protect public health by controlling the safety and efficacy of merchandise in areas including pharmaceuticals, complementary color medicines, veterinary medicines, medical devices, and even food products and cosmetics. Method: Herbal health products are in practices in all parts of the world under either their legal system's or expert council's or agencies' guides. They include botanicals, health supplements, health foods, complimentary medicines, traditional medicines or following pathies like Ayurveda, Yoga, Unani, Siddha, and Homeopathy. The requisite parameters for registration or recognition of products by various major global regulatory agencies were reviewed, and compiled under purview. Results: In India, licensing these products is under the act provisions and the rules known as the Drugs and Cosmetics Act, whereas globally regulatory provisions follow the guidelines of developed countries like the U.S. Food and Drug Administration, European Medicines Agency, the Therapeutic Goods Act, or the World Health Organization's regulations for herbal products. Conclusions: The present communication highlights the provisions of regulatory and/or licensing requirements related to corporates, product composition, specifications, quality parameters, manufacturing methodology, stability, safety, preclinical studies, clinical studies, etc. for herbal products and the respective guidelines at one site. Highlights: Ultimately, all regulatory agencies across the world highlight majorly the safety and thereafter the efficacy for any products under the category.

The role of the South African Medical Research Council in reducing lead exposure and preventing lead poisoning in South Africa.

Even at low levels in blood, lead has been associated with reduced IQ scores, behavioural problems, learning impediments, aggression and violent behaviour. Since the 1980s, the South African Medical Research Council (SAMRC) has been investigating the sources of exposure to lead in South Africa (SA), the groups at highest risk of lead poisoning and a selection of the myriad associated health and social consequences. SAMRC research evidence contributed to the phasng out of leaded petrol, restrictions on lead in paint and other interventions. Subsequently, childhood blood lead levels in SA declined significantly. More recent studies have revealed elevated risks of lead exposure in subsistence fishing and mining communities, users of arms and ammunition, those ingesting certain traditional medicines, and users of certain ceramicware and artisanal cooking pots. Lead-related cognitive damage costs the SA economy ~USD17.7 (ZAR261.3) billion annually, justifying further SAMRC investment in lead exposure research in the country.

Ethanol content in traditionally fermented ayurvedic formulations: Compromised Good Manufacturing Practice regulations - compromised health.

BACKGROUND: A large part of the population of India prefers the traditional medicine (ayurvedic formulations) for primary health care. However, the effective quality control of herbal medicine is still a big challenge. Numerous reports indicate noncompliance with Compromised Good Manufacturing Practice (GMP) guidelines by the manufacturers which may lead to adverse drug reactions or toxic effects. Asava and arishta are the classical herbal dosage forms wherein fermentation occurs during production leading to the generation of ethanol. The presence of ethanol in these preparations may lead to their misuse. The self-generated ethanol is responsible for extraction of active constituents and acts as a self-preservative. As the procedure for preparation for asava and arishta is same, the ethanol content is also expected to be the same irrespective of the manufacturer. OBJECTIVE: The objective of the present study was to assess and compare the ethanol content of some traditionally fermented ayurvedic formulations available in the market. METHOD: In this study, 20 formulations from 3 different manufacturers available as over-the-counter products were obtained and their ethanol contents were determined using gas chromatograph with flame ionization detector. RESULTS: Statistically significant differences were noted in the ethanol content of various ayurvedic formulations. A simple, less time-consuming, economic, and validated gas chromatographic method for estimation of ethanol in fermented ayurvedic formulations was also developed successfully in present study. CONCLUSION: The data generated during study reflected poor compliance of GMP guidelines by the manufacturers and hence the quality is being grossly compromised posing a safety hazard.

Can the AYUSH system be instrumental in achieving universal health coverage in India?

Universal health coverage (UHC) in the Indian context is understood as easily accessible and affordable health services for all citizens. The Planning Commission of India constituted a High Level Expert Group (HLEG) in October 2010 for the purpose of drafting the guidelines of UHC. While the primary focus of UHC is to provide financial protection to all citizens, its delivery requires an adequate health infrastructure, skilled health human resources, and access to affordable drugs and technologies so that all people receive the level and quality of care they are entitled to. This paper attempts to link the ayurveda, yoga and naturopathy, unani, siddha and homoeopathy (AYUSH) systems of medicine with UHC. Here, the AYUSH system refers to the AYUSH workforce, therapeutics and principles, and their individual role in delivering UHC to the citizens of India. In outlining the role of AYUSH, the paper lays stress on the 10 guiding principles of UHC, as proposed by the HLEG. However, as the AYUSH system is not the principal health service provider in India, the dominant system being that of allopathic medicine, a few components of UHC may not fit neatly into the AYUSH system. This paper has adopted the definition of UHC quoted by the HLEG.

Preclinical renal chemo-protective potential of Prunus amygdalus Batsch seed coat via alteration of multiple molecular pathways.

Prunus amygdalus Batsch (almond) is a classical nutritive traditional Indian medicine. Along with nutritive with anti-oxidant properties, it is, clinically, used in the treatment of various diseases with underlying anti-oxidant mechanism. This study is an effort to scrutinise the renal protective effect of P. amygdalus Batsch or green almond (GA) seed coat extract and its underlying mechanism in animal model of Ferric nitrilotriacetate (Fe-NTA) induced renal cell carcinoma (RCC). RCC was induced in Swiss Albino Wistar rats by intraperitoneal injection of Fe-NTA. The rats were then treated with ethanolic extract of GA (25, 50 and 100 mg/kg per oral) for 22 weeks. Efficacy of GA administration was evaluated by change in biochemical, renal, macroscopical and histopathological parameters and alterations. Additionally, interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and inflammatory mediator including prostaglandin E2 (PGE2), nuclear factor-kappa B (NF-κB) were also observed to explore the possible mechanisms. The oral administration of GA significantly (p < .001) altered the Fe-NTA induced RCC in rats by inhibition of renal nodules, decolourisation of tissues, tumour promoter marker including thymidine 3[H] incorporation, ornithine decarboxylase, renal parameters and anti-oxidant parameters in serum. Additionally, GA treatment significantly (p < .001) down-regulated the IL-6, IL-1ß, TNF-α, inflammatory mediators PGE2 and NF-κB in a dose-dependent manner. Histopathology observation supported the renal protective effect of GA by alteration in necrosis, size of Bowman capsules and inflammatory cells. Hence, it can be concluded that GA possesses observable chemo-protective action and effect on Fe-NTA induced RCC via dual inhibition mechanism one by inhibiting free radical generation and second by inhibiting inflammation.