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1.  Development of a RP-HPLC Method for Analysis of Triphala Curna and its Applicability to Test Variations in Triphala Curna Preparations 
A sensitive, rapid, reverse phase HPLC method is reported for analysis of Triphala Curna using gallic acid, chebulagic acid and chebulinic acid as markers. Validation data for the method has been provided. Unlike methods of recovery testing adopted for synthetic chemicals, a modified approach has been presented here for a formulation like Triphala Curna having three myrobalans in its composition. Data has been provided to demonstrate applicability of the method developed to assess the variation in the Triphala Curna preparations.
doi:10.4103/0250-474X.57286
PMCID: PMC2865809  PMID: 20502543
Triphala Curna (TC); gallic acid; chebulagic acid; chebulinic acid; myrobalans
2.  OA01. 37. Clinical evaluation of rasayana effect of ashwagandhadi lehya in apparently healthy elderly persons 
Ancient Science of Life  2012;32(Suppl 1):S37.
Purpose:
This study has been conducted on 50 apparently healthy elderly persons by treating them with Ashwagandhadi Lehya to assess the efficacy and safety of the drug on various health parameters.
Method:
50 patients of age between 50 -75 years were registered from the O.P.D. of N.I.A., Jaipur in this study out of which total 42 patients completed treatment. Ashwagandhadi Lehya was given in the dose of 10 gm BD After meals with Luke warm milk for 84 days. Diet restrictions according to Pathya -Apathya were advised.
Result:
This formulation provided statistically highly significant relief in chief complaints like dizziness, constipation, abnormal sleep, loss of appetite, fatigue, generalized weakness, sense of well being and in mental aspect depression, physical and psychological health also. The drug was effective to improve Hb%, change in serum creatinine, unconjugated bilirubin, serum cholesterol and LDL levels significantly.
Conclusion:
A number of Medhya Rasayana Dravyas present in formulation have very good anti oxidant, anabolic, adaptogenic, anti stress and anti depressant property that improves the mental and physical health of the aged person.
PMCID: PMC3800915
3.  STUDIES ON THE STANDARDISATION OF CURNAS PART – II TALIZADYA CURNA 
Ancient Science of Life  1991;11(1-2):46-49.
Talisadya churna was prepared by pounding the individual ingredients in mortar and pestle and mixie. The Curna prepared by pounding the ingredients in mortar and pestle showed higher exhaustive extraction in hexane and solubility in alcohol. The Curna prepared by grinding the ingredients in mixie showed less acid insoluble content, high volatile matter, water soluble matter, and exhaustive extraction in chloroform. Thin layer silica gel chromatography and test of organic functional groups did not show any difference in the Tulisadya curna prepared by either method.
PMCID: PMC3336572  PMID: 22556560
4.  EFFECT OF CONTAINER ON AYURVEDIC DRUGS – A SELECT STUDY 
Ancient Science of Life  2004;24(1):11-21.
Effect of packing material on the stability of four Ayurvedic drugs viz., Hingvatsaka Curna, Brahmi ghrta, Dasamula Kvatha Curna and Ajamodarka have been studied by distributing the drugs in different containers by observing the physical and biological changes. The study revealed that Hingvatsaka Curna keeps well for at least 6 month, Ajamodarka is stable for minimum period of six months when stored in glass bottle exposed to light. Dasamula K vatha Curna showed the presence of insects at the completion of three months. In the second month itself Brahmi ghrta becomes rancid on storing in glass and amber colored bottles.
PMCID: PMC3330913  PMID: 22557145
Container effect; Hingvatsaka Curna; Brahmi ghrta; Dasamula Kvatha Curna; Ajamodarka
5.  Effects of Ayurvedic treatment on 100 patients of chronic renal failure (other than diabetic nephropathy) 
Ayu  2011;32(4):483-486.
Chronic renal failure (CRF) refers to an irreversible deterioration in renal function, which develops over a period of years. This initially manifests only as a biochemical abnormality. CRF is considered when glomerular filtration rate (GFR) falls below 30 ml/min. The conventional approach of management includes dialysis and renal transplantation, which are not affordable by Indian population mainly due to economic reasons. Therefore, exploration of a safe and alternative therapy is needed, which proves to be helpful in reducing the requirement of dialysis and in postponing the renal transplantation. A clinical study of 100 patients of CRF was conducted at OPD and IPD of PD Patel Ayurved Hospital, Nadiad. They were given Niruha basti of Punarnavadi kvatha daily with oral medicaments including Goksuradi guggulu, Rasayana churna, and Varunadi kvatha for 1 month period. The patients of CRF, having diabetic nephropathy as a cause, were excluded since a separate study for diabetic nephropathy is being conducted. Results were analyzed statistically using the “t” test. The symptoms and signs, serum creatinine, blood urea, urine albumin level were reduced, which were found to be statistically highly significant on “t” test.
doi:10.4103/0974-8520.96120
PMCID: PMC3361922  PMID: 22661841
Ayurveda; chronic renal failure; Niruha basti
6.  A CLINICAL STUDY ON THE MANAGEMENT OF DEPRESSIVE NEUROSIS WITH RASAYANA - VAJIKARNA DRUGS 
Ancient Science of Life  1983;2(4):220-226.
In view of the potential anti-anxiety and anti–depressant activity of the Rasayana Vajikarna drugs, a combined Rasayana Vajikarna schedule has been clinically tried in a series of patients of Depression Neurosis. The tratment consisted of a dose of Asvagandha Curna at bed time and a dose of Kapikacchu Curna at morning continued for 8 weeks. The treatment showed significant clinical recovery in these cases in the form of a notable reduction in the level of clinical anxiety and depression.
PMCID: PMC3336761  PMID: 22556986
7.  Scientific evaluation of some Ayurvedic preparations for correction of iron deficiency and anemia 
A number of preparations are available in Ayurved for treatment of anemia and iron deficiency. This study was designed to evaluate efficacy of some of them. Six most commonly used Ayurvedic iron containing preparations (Navayasa Curna, Punarnavadi Mandura, Dhatri Lauha, Pradarantaka Lauha, Sarva-Juara-Hara Lauha and Vrihat Yakrdari Lauha) were given in a dose of 250 mg b.d. for 30 days to six groups of iron deficient anemic patients; each group consisting of 20 patients. A control group was given Allopathic preparation—Irex-12, (containing—ferrous fumarate, vitamin C, folic acid and vitamin B12); 1 capsule daily for 30 days for comparison. All hematological and iron parameters were determined before and after completion of treatment. The results showed that there was statistically significant rise (p<0.001) in all of them—Hb, PCV, TRBC, MCV, MCH, MCHC and plasma iron, percent saturation and plasma ferritin. Total iron binding capacity decreased significantly (p<0.001). The response of most of Ayurvedic preparations was better than Allopathic preparation and there was no side effect as observed with iron salts The Hb regeneration rate was 0.10 g/dl/day for Allopathic preparation; while it was above this value for all Ayurvedic preparations exceptPradarantaka Lauha which was least effective.Sarva-Juara-Hara Lauha was the drug of choice as Hb regeneration with it was highest 0.16 g/dl/day. Upon analysis of Ayurvedic drugs, these results were found to be consistent and correlated with iron content of the preparation.
doi:10.1007/BF02913329
PMCID: PMC3453821  PMID: 23105698
Ayurvedic preparations; Iron deficiency; Anemia; Hematological parameters; Iron parameters
8.  PHARMACOGNOSTICAL INVESTIGATIONS ON TRIPHALA CHURNAM 
Ancient Science of Life  2007;26(3):40-44.
Pharmacognostical and preliminary phytochemical studies of Triphala churnam were carried out. The churnam of triphala consists of equal quantities of deseeded fruits of Terminalia chebula, Terminalia bellerica and Emblica officinalis. Triphala is exclusively used in more than 200 drug formulations in Indian system of Medicine. The present study involved the pharmacognostical evaluation of Triphala, in which morphological and powder microscopical characters were established. In addition, physico-chemical parameters such as ash values viz, total ash (10.21± 0.42), acid insoluble ash (2.54 ± 0.06), water-soluble ash (5.46±0.24) and sulphated ash (13.12 ± 0.63), extractive values viz, alcohol soluble extractive (11.20±0.18)) and water-soluble extractive (52.56±2.04), fluorescent analysis and microchmical tests were determined. The preliminary phytochemical study revealed the presence of carbohydrates, reducing sugar and tannins in aqueous extract and carbohydrates, flavonoids and tannins in alcoholic extract. This standardization would be very much helpful for the identification of Triphala churnam to differentiate from other powdered sources.
PMCID: PMC3330880  PMID: 22557240
9.  Studies on Brahma rasayana in male swiss albino mice: Chromosomal aberrations and sperm abnormalities 
Ayurveda, the Indian holistic healthcare system encompasses traditional medicines with a principle of creating harmony and maintaining balance within the natural rhythms of the body. Rasayana is one of the branches of Ayurveda frequently used as rejuvenant therapy to overcome many discomforts and prevent diseases. It has been reported that rasayanas have immunomodulatory, antioxidant and antitumor functions. However, the genotoxic potential of many rasayanas remains to be evaluated. The present study was undertaken to assess the role of Brahma rasayana(BR) on genotoxicity in vivo in a mouse test system. The older mice (9 months) were orally fed with rasayana for 8 weeks. The treated groups showed no signs of dose-dependent toxicity at the dosage levels tested. The body weight loss/gain and feed consumption were unaffected at tested doses. Furthermore, sperm abnormalities and chromosomal aberrations were insignificant in the treatment group when compared to controls. However, there was a marginal increase in sperm count in the BR treated animals. These findings clearly indicate that there are no observed adverse genotoxic effects elicited by BR in experimental animals such as mice.
doi:10.4103/0975-9476.59826
PMCID: PMC3149391  PMID: 21829300
Aging; Brahma rasayana; chromosomal aberrations; genotoxicity; sperm abnormalities
10.  Exploring of Antimicrobial Activity of Triphala Mashi—an Ayurvedic Formulation 
Triphala Mashi is an ayurvedic formulation that was prepared in our lab. Aqueous and alcoholic extracts of both Triphala and Triphala Mashi were used, to evaluate antimicrobial activity. Comparative phytochemical profile of Triphala and Triphala Mashi was done by preliminary phytochemical screening, total phenolic content and thin layer chromatography (TLC). Antimicrobial activity includes isolation of pathogens from clinical samples, its characterization, testing its multiple drug resistance against standard antibiotics and antimicrobial activity of aqueous and alcoholic extracts of both Triphala and Triphala Mashi against these organisms by using agar gel diffusion method. Triphala Mashi containing phenolic compounds, tannins exhibited comparable antimicrobial activity in relation to Triphala against all the microorganisms tested. It inhibits the dose-dependent growth of Gram-positive and Gram-negative bacteria. In conclusion, it appears that Triphala Mashi has non-specific antimicrobial activity.
doi:10.1093/ecam/nem002
PMCID: PMC2249739  PMID: 18317557
antimicrobial activity; clinical sample and Tannin; Triphala; Triphala Mashi
11.  Brahmarasayana protects against Ethyl methanesulfonate or Methyl methanesulfonate induced chromosomal aberrations in mouse bone marrow cells 
Background
Ayurveda, the traditional Indian system of medicine has given great emphasis to the promotion of health. Rasayana is one of the eight branches of Ayurveda which refers to rejuvenant therapy. It has been reported that rasayanas have immuno-modulatory, antioxidant and antitumor functions, however, the genotoxic potential and modulation of DNA repair of many rasayanas have not been evaluated.
Methods
The present study assessed the role of Brahmarasayana (BR) on Ethyl methanesulfonate (EMS)-and Methyl methanesulfonate (MMS)-induced genotoxicity and DNA repair in in vivo mouse test system. The mice were orally fed with BR (5 g or 8 mg / day) for two months and 24 h later EMS or MMS was given intraperitoneally. The genotoxicity was analyzed by chromosomal aberrations, sperm count, and sperm abnormalities.
Results
The results have revealed that BR did not induce significant chromosomal aberrations when compared to that of the control animals (p >0.05). On the other hand, the frequencies of chromosomal aberrations induced by EMS (240 mg / kg body weight) or MMS (125 mg / kg body weight) were significantly higher (p<0.05) to that of the control group. The treatment of BR for 60 days and single dose of EMS or MMS on day 61, resulted in significant (p <0.05) reduction in the frequency of chromosomal aberrations in comparison to EMS or MMS treatment alone, indicating a protective effect of BR. Constitutive base excision repair capacity was also increased in BR treated animals.
Conclusion
The effect of BR, as it relates to antioxidant activity was not evident in liver tissue however rasayana treatment was observed to increase constitutive DNA base excision repair and reduce clastogenicity. Whilst, the molecular mechanisms of such repair need further exploration, this is the first report to demonstrate these effects and provides further evidence for the role of brahmarasayana in the possible improvement of quality of life.
doi:10.1186/1472-6882-12-113
PMCID: PMC3457898  PMID: 22853637
DNA damage and repair; Anti clastogenicity; Rasayana
12.  A clinical study of Haritaki and Saindhava Lavana in Kaphaja Kasa with special reference to Samyoga Guna 
Ayu  2011;32(3):357-360.
In clinical practice, Guna which are to be with Bhisak are mainly the Paradi Gunas which can also be called as Miscellaneous Gunas. As rightly quoted by Acarya Caraka, for getting success in the treatment Paradi Gunas are the best. The Sutra quotes “Sidhyupaya Cikitsayam” which means that Cikitsa i. e. Dhatusamya will be done mainly with the help of Paradi Gunas. Thus in this study an attempt was made to know the comparative effect of Haritaki and Saindhava lavana alone and Samyukta effect in Kaphaja Kasa. Three groups were made for proper evaluation of the therapy. In Group-A Haritaki Tablet 2 gm/ twice a day (500 mg tablet × 4), In Group-B Saindava Lavana Curna 2 gm/ twice a day and in Group-C Haritaki + Saindhava lavana Tablet 4 gm/twice a day (500 mg tablet × 8) was given. Results were assessed after 7 days with the help of a specially prepared proforma. The hematological, Urine and Stool investigations were carried out. In subjective and objective criterias, significant results were found in Group-C as compared to Group-A and Group-B. Based on the results, it can be concluded that the combined (Samyoga) effect of Haritaki and Saindhava lavana is much efficient than the single drug therapy.
doi:10.4103/0974-8520.93914
PMCID: PMC3326882  PMID: 22736910
Guna; Haritaki; Paradi Guna; Saindhava lavana; Samyoga
13.  Significant Increase in Cytotoxic T Lymphocytes and Natural Killer Cells by Triphala: A Clinical Phase I Study 
Background. Searching for drugs or herbal formulations to improve the immunity of HIV/AIDS positive people is an important issue for researchers in this field. Triphala, a Thai herbal formulation, is reported to have immunomodulatory effects in mice. However, it has not yet been investigated for immunostimulatory and side effects in healthy human volunteers. Objective. To evaluate the immunostimulatory and side effects of Triphala in a clinical phase I study. Materials and Methods. All volunteers took Triphala, 3 capsules per day for 2 weeks. Complete physical examination, routine laboratory analysis, and immunological studies were performed before ingestion and after initial meeting for 4 consecutive weeks. Results. We found that Triphala demonstrated significant immunostimulatory effects on cytotoxic T cells (CD3−CD8+) and natural killer cells (CD16+CD56+). Both of them increased significantly when compared with those of the control samples. However, no significant change in cytokine secretion was detected. All volunteers were healthy and showed no adverse effects throughout the duration of the study. Conclusion. Triphala has significant immunostimulatory effects on cellular immune response, especially cytotoxic T cells and natural killer cells. Increases in the absolute number of these cells may provide a novel adjuvant therapy for HIV/AIDS positive people in terms of immunological improvement.
doi:10.1155/2012/239856
PMCID: PMC3519011  PMID: 23243435
14.  Enhancement of basolateral amygdaloid neuronal dendritic arborization following Bacopa monniera extract treatment in adult rats 
Clinics  2011;66(4):663-671.
OBJECTIVE:
In the ancient Indian system of medicine, Ayurveda, Bacopa monniera is classified as Medhya rasayana, which includes medicinal plants that rejuvenate intellect and memory. Here, we investigated the effect of a standardized extract of Bacopa monniera on the dendritic morphology of neurons in the basolateral amygdala, a region that is concerned with learning and memory.
METHODS:
The present study was conducted on 2½-month-old Wistar rats. The rats were divided into 2-, 4- and 6-week treatment groups. Rats in each of these groups were further divided into 20 mg/kg, 40 mg/kg and 80 mg/kg dose groups (n  =  8 for each dose). After the treatment period, treated rats and age-matched control rats were subjected to spatial learning (T-maze) and passive avoidance tests. Subsequently, these rats were killed by decapitation, the brains were removed, and the amygdaloid neurons were impregnated with silver nitrate (Golgi staining). Basolateral amygdaloid neurons were traced using camera lucida, and dendritic branching points (a measure of dendritic arborization) and dendritic intersections (a measure of dendritic length) were quantified. These data were compared with the data from the age-matched control rats.
RESULTS:
The results showed an improvement in spatial learning performance and enhanced memory retention in rats treated with Bacopa monniera extract. Furthermore, a significant increase in dendritic length and the number of dendritic branching points was observed along the length of the dendrites of the basolateral amygdaloid neurons of rats treated with 40 mg/kg and 80 mg/kg of Bacopa monniera (BM) for longer periods of time (i.e., 4 and 6 weeks).
CONCLUSION:
We conclude that constituents present in Bacopa monniera extract have neuronal dendritic growth-stimulating properties.
doi:10.1590/S1807-59322011000400023
PMCID: PMC3093798  PMID: 21655763
Bacopa monniera; spatial learning; passive avoidance; amygdaloid neurons; dendritic arborization; memory
15.  Triphala inhibits both in vitro and in vivo xenograft growth of pancreatic tumor cells by inducing apoptosis 
BMC Cancer  2008;8:294.
Background
Triphala is commonly used in Ayurvedic medicine to treat variety of diseases; however its mechanism of action remains unexplored. This study elucidates the molecular mechanism of Triphala against human pancreatic cancer in the cellular and in vivo model.
Methods
Growth-inhibitory effects of Triphala were evaluated in Capan-2, BxPC-3 and HPDE-6 cells by Sulphoradamine-B assay. Apoptosis was determined by cell death assay and western blotting. Triphala was administered orally to nude mice implanted with Capan-2 xenograft. Tumors were analyzed by immunohistochemistry and western blotting.
Results
Exposure of Capan-2 cells to the aqueous extract of Triphala for 24 h resulted in the significant decrease in the survival of cells in a dose-dependent manner with an IC50 of about 50 μg/ml. Triphala-mediated reduced cell survival correlated with induction of apoptosis, which was associated with reactive oxygen species (ROS) generation. Triphala-induced apoptosis was linked with phosphorylation of p53 at Ser-15 and ERK at Thr-202/Tyr-204 in Capan-2 cells. Above mentioned effects were significantly blocked when the cells were pretreated with an antioxidant N-acetylcysteine (NAC), suggesting the involvement of ROS generation. Pretreatment of cells with pifithrin-α or U0126, specific inhibitors of p53 or MEK-1/2, significantly attenuated Triphala-induced apoptosis. Moreover, NAC or U0126 pretreatment significantly attenuated Triphala-induced p53 transcriptional activity. Similarly, Triphala induced apoptosis in another pancreatic cancer cell line BxPC-3 by activating ERK. On the other hand, Triphala failed to induce apoptosis or activate ERK or p53 in normal human pancreatic ductal epithelial (HPDE-6) cells. Further, oral administration of 50 mg/kg or 100 mg/kg Triphala in PBS, 5 days/week significantly suppressed the growth of Capan-2 pancreatic tumor-xenograft. Reduced tumor-growth in Triphala fed mice was due to increased apoptosis in the tumors cells, which was associated with increased activation of p53 and ERK.
Conclusion
Our preclinical studies demonstrate that Triphala is effective in inhibiting the growth of human pancreatic cancer cells in both cellular and in vivo model. Our data also suggests that the growth inhibitory effects of Triphala is mediated by the activation of ERK and p53 and shows potential for the treatment and/or prevention of human pancreatic cancer.
doi:10.1186/1471-2407-8-294
PMCID: PMC2576337  PMID: 18847491
16.  A Metabolomics-Based Strategy for the Quality Control of Traditional Chinese Medicine: Shengmai Injection as a Case Study 
Quality control of traditional Chinese medicines (TCMs) used clinically is becoming a challenge and has limited the development of TCM due to the high variability in concentration levels of active ingredients and markers as well as the lack of well-established criteria. Using Shengmai injection, which is a well-established TCM, as an example, we developed an integrated profiling approach that simultaneously captures the entire spectrum of ingredients and quantitatively determines the levels of seven key ingredients in the TCM product. A multivariate statistical model was constructed to establish a “seven-marker-” based quality standard that qualified the majority of samples in this study. This newly developed strategy showed that a panel of key ingredients or markers in the TCM product were relatively consistent within a statistically acceptable range. Therefore, this metabolomics-based approach will complement the current quality control standard using the concentration of several key ingredients or their total content and help improve the consistency and clinic efficacy of TCM products.
doi:10.1155/2013/836179
PMCID: PMC3562660  PMID: 23401718
17.  Quantification of Gallic Acidin Fruits of Three Medicinal Plants 
Triphala is a traditional herbal formulation consisting of dried fruits originating from three medicinal plants, namely Terminalia chebula, Terminalia bellerica and Phyllanthus emblica. It is used in folk medicine for the treatment of headaches, dyspepsia and leucorrhoea. There are some reports regarding Triphala’s pharmacological effects including its anti-cancer, radioprotective, hypocholesterolaemic, hepatoprotective and anti-oxidant activities. The most important components of these plants are the tannins and gallic acid which they contain. Gallic acid being a compound with tannin structure existing in the Triphala fruit. In this research, the gallic acid content contained in the three plants constituting Triphala was determined. Plant fruits were purchased from available Iranian markets. Milled and powdered fruits from each plant were extracted with 70% acetone and subjected to a reaction with rhodanine reagent in the process forming a colored complex. The complex’s absorbance was measured at 520 nm and the amount of gallic acid was determined using its calibration curve. According to the results, the highest amount of gallic acid was observed in Phyllanthus embelica (1.79-2.18%) and the lowest amount was found in Terminalia chebula (0.28-0.80%). Moreover, differences between plant samples from different markets places were found to be statistically significant (p < 0.05). These differences can possibly be due to the source of plant preparation, storage condition and period of Triphala storage. In general, the rhodanine assay is a simple, rapid and reproducible method for the standardization of Triphala as gallic acid.
PMCID: PMC3828909  PMID: 24250348
Triphala; Terminalia chebula; Terminalia bellerica; Phyllanthus emblica; Gallic acid; Rhodanine assay
18.  RAPD Analysis for Determination of Components in Herbal Medicine 
In this study, the RAPD (Random Amplified Polymorphic DNA) technique was employed for determination of the components in an Ayurvedic herbal prescription, Rasayana Churna. One-hundred-and-twenty decamer oligonucleotide primers were screened in the RAPD analysis to identify three Ayurvedic medicines, dried stem of Tinospora cordifolia, dried fruit of Emblica officinalis and dried fruit of Tribulus terestris, the Ayurvedic prescription. Primer OPC-6 simultaneously generated three distinct amplicons, each specific to one component. The marker with 600 bp is specific to Tinospora cordifolia; the marker 500 bp is specific to Emblica officinalis and the remaining marker >1000 bp was present in Tribulus terestris. Presence of three herbal medicines was determined when RAPD reaction with OPC-6 was performed. The technique was proved to contribute to the identification of components in Ayurvedic herbal preparation and thus helping to serve as a complementary tool for quality control.
doi:10.1093/ecam/nem109
PMCID: PMC2206231  PMID: 18227927
DNA fingerprinting; herbal medicine; RAPD; standardization
19.  Increase in Drosophila melanogaster longevity due to rasayana diet: Preliminary results 
We report preliminary results from an ongoing series of experiments on lifespan extension by appropriately modified Ayurvedic rasayanas in animal models. Here data are presented indicating lifespan extensions of 51–55% (up to 70–95% in the pilot experiment) in a standard strain of Drosophila melanogaster (Oregon-K) using a standard rasayana (Ayurvedic herbal formulation for life-extension) suitably adapted for insects. In a first experiment, two groups of 20 unmated D. melanogaster strain Oregon-K kept at 22°C received either rasayana or standard yeast diet; days of death were recorded. Another experiment investigated possible sex differences; equal sized (N = 30) groups of similar males, females, and controls were compared. Life lengths of all controls were in the strain's usual range: in Experiment 1, control life lengths were minimum 40 to maximum 53 days; experimental group figures were 81–91 days; groups were completely separated, experimental group minimum life length being 28 days more than control group maximum life length, i.e., about 2.5 full distribution widths – a sign test for the null hypothesis yields p < 2–20, i.e., 10–6 as maximum p. Experiment 2 found no differences between life lengths of males and females; but the maximum life length of 30 controls (60 days in males and 66 days in females) was once again far shorter than the minimum life length of the 60 in the two experimental groups, strengthening the findings of Experiment 1. Despite group sizes being relatively small, results are conclusive: the rasayanas in question increase D. melanogaster strain Oregon-K life length. The complexity of the formulation suggests that multiple mechanisms are involved – worth further investigation.
doi:10.4103/0975-9476.65085
PMCID: PMC3151378  PMID: 21836798
Ayurveda; rasayana; D. melanogaster; longevity
20.  Standardization of a Traditional Polyherbo-Mineral Formulation-Brahmi Vati 
The present study deals with standardization of an in-house standard preparation and three marketed samples of Brahmi vati, which is a traditional medicine known to be effective in mental disorders, convulsions, weak memory, high fever and hysteria. Preparation and standardization have been done by following modern scientific quality control procedures for raw material and the finished products. The scanning electron microscopic (SEM) analysis showed the reduction of metals and minerals (particle size range 2–5 µm) which indicates the proper preparation of bhasmas, the important ingredient of Brahmi vati. Findings of EDX analysis of all samples of Brahmi vati suggested the absence of Gold, an important constituent of Brahmi vati in two marketed samples. All the samples of Brahmi vati were subjected to quantitative estimation of Bacoside A (marker compound) by HPTLC technique. Extraction of the samples was done in methanol and the chromatograms were developed in Butanol: Glacial acetic acid: water (4.5:0.5:5 v/v) and detected at 225nm. The regression analysis of calibration plots of Bacoside A exhibited linear relationship in the concentration range of 50–300 ng, while the % recovery was found to be 96.06% w/w, thus proving the accuracy and precision of the analysis. The Bacoside A content in the in-house preparation was found to be higher than that of the commercial samples. The proposed HPTLC method was found to be rapid, simple and accurate for quantitative estimation of Bacoside A in different formulations. The results of this study could be used as a model data in the standardization of Brahmi vati.
PMCID: PMC3777576  PMID: 24146464
Brahmi vati(BV); Standardization; EDX; HPTLC; Bacoside-A
21.  Quality control of tuirejieduling granules using high-performance liquid chromatography fingerprint method and simultaneous determination of four main active ingredients 
Pharmacognosy Magazine  2013;9(34):162-166.
Background:
The Tuirejieduling granule is a compound preparation made from four kinds of Chinese medicines. It is effective for anti-inflammation, antivirus, defervescence and anti-bacterium; however, its quality control standards have remained unknown.
Objective:
To establish a simple and accurate fingerprint method for quality control of the Tuirejieduling granule.
Materials and Methods:
The methanol extract of the Tuirejieduling granule was used for the fingerprint analysis and the four selected active ingredients (epigoitrin, phillyrin, saikosaponin A and glycyrrhetinic acid) in the extract were determined. The fingerprint method was performed on an Amethyst C18-P chromatography column by gradient elution with acetonitrile and aqueous phase (containing 0.5% H3PO4 (v/v), pH 3.0).
Results:
Under the optimal chromatographic condition, twenty peaks were chosen as fingerprint peaks of the Tuirejieduling granules extractions. The similarities of 10 batches of Tuirejieduling granule was more than 0.99. This indicates that the different batches of Tuirejieduling granules were under the consistent quality control. Good linear behaviors over the investigated concentration ranges were obtained with the values of R2 higher than 0.99 for four studied active ingredients. The recoveries for spiked samples were in the range of 96.2–105.5%. The developed method was successfully applied to determine the contents of active constituents in different batches of Tuirejieduling granule.
Conclusion:
The HPLC fingerprint was proved to be a reliable method for the quality control of Tuirejieduling granule.
doi:10.4103/0973-1296.111284
PMCID: PMC3680857  PMID: 23772113
Fingerprint; high-performance liquid chromatography; quality control; Tuirejieduling granules
22.  Efficacy of ‘Itrifal Saghir’, a combination of three medicinal plants in the treatment of obesity; A randomized controlled trial 
Background
Herbal combination of Itrifal Saghir (triphala) has been widely used in traditional medicine. And brings health benefits such as antioxidant effect and scavenger of hydroxyl radicals and nitric oxide radicals activity and substantiated in traditional medicine a anti-obesity.
Material and method
In this study we aimed to assess the efficacy of this herbal medicinal on reduction of weight and body mass index (BMI) of simple obese subjects in comparison with placebo. Obese subjects aged between 16 and 60 years were selected for 12-week, double-blind, randomized, placebo-controlled trial using a parallel design. Subjects were randomly assigned to take 5 grams of either the Itrifal Saghir (n = 31) or placebo (n = 31), 2 times daily for 12 weeks. Measures of body weight, BMI, waist circumference (WC), hip circumference (HC), were assessed at baseline and once every four weeks during the 12 week treatment period. The safety was evaluated by means of measuring the liver and kidney function. Homeostasis model of insulin resistance (HOMA-IR) was calculated as [fasting insulin (μU/mL) × fasting glucose (mmol/L)/22.5].
Results
Compared to placebo group, in treatment group the mean difference of effective weight loss was 4.82Kg (CI95% 3.52 - 6.11, ρ < 0.001), the mean of decrease in waist circumference was 4.01 cm (CI 95% 2.13 - 5.90, ρ < 0.001), and the mean decrease in hip circumference was 3. 21 cm (CI 95% 1.96 - 4.45, ρ < 0.001) in treated subjects. No adverse effects or significant changes in liver and kidney function tests were observed in both placebo and treated groups.
Conclusions
Itrifal Saghir appears to produce a positive effect on weight loss in obese subjects.
doi:10.1186/2008-2231-20-33
PMCID: PMC3559014  PMID: 23351558
Obesity; Itrifal Saghir; Traditional Iranian medicine; Randomized trial; Terminalia chebula; Terminalia bellerica; Emblica officinalis; Triphala
23.  STANDARDISATION OF “HAB – E – BUKHAR” (Unani Medicine) 
Ancient Science of Life  1985;5(1):37-39.
‘HAB-E-BUKHAR’ – a compound drug formulation in Unani System of medicine has been analysed. The microscopic method of identification of few of its ingredients (Sath-e-gulu, Thabasheer); Quantitative analysis of Quinine sulphate, other physio-chemical contents and the T.L.C. of the drug have also been reported here.
PMCID: PMC3331439  PMID: 22557497
24.  Terpinen-4-ol is the Most Active Ingredient of Tea Tree Oil to Kill Demodex Mites 
Purpose
To determine the active ingredient in tea tree oil (TTO) responsible for its reported killing effect on Demodex mites, the most common ectoparasite found in the human skin extending to the eye.
Methods
Using a reported in vitro killing assay to measure the survival time of adult Demodex folliculorum up to 150 minutes, we have screened serial concentrations of 13 of the 15 known ingredients of TTO (ISO4730:2004) that were soluble in mineral oil and examined their synergistic relationships in killing mites. The most potent ingredient was then tested for its efficacy in killing Demodex in vivo.
Results
All ingredients exhibited a dose-dependent killing effect. Besides Terpinen-4-ol, the order of relative potency did not correlate with the order of relative abundance in TTO for the remaining 12 ingredients. Terpinen-4-ol was the most potent ingredient followed by α-Terpineol, 1,8-Cineole and Sabinene. Terpinen-4-ol, the most abundant ingredient in TTO, was more potent than TTO at equivalent concentrations and its killing effect was even observable at a mere concentration of 1%. Terpinen-4-ol exhibited a significant synergistic effect with Terpinolene, but an antagonistic effect with α-Terpineol in killing mites (both P < 0.05). In vivo, Terpinen-4-ol was shown to eradicate mites.
Conclusions
The above finding suggests that deployment of Terpinen-4-ol alone should enhance its potency in killing Demodex mites by reducing the adverse and antagonistic effects from other ingredients in TTO.
Translational Relevance
Terpinen-4-ol can be adopted in future formulations of acaricides to treat a number of ocular and cutaneous diseases caused by demodicosis.
doi:10.1167/tvst.2.7.2
PMCID: PMC3860352  PMID: 24349880
terpinen-4-ol; tea tree oil; demodex; acaricide; ocular drug delivery
25.  Development of ITS sequence-based markers to distinguish Berberis aristata DC. from B. lycium Royle and B. asiatica Roxb. 
3 Biotech  2011;1(1):11-19.
The stems of Berberis aristata DC. (Berberidaceae) are used in the South Asian traditional medicine as a key ingredient in formulations for eye care, skin diseases, jaundice, rheumatism and also in diabetes. B. lycium Royle and B. asiatica Roxb. are traded as equivalents of B. aristata. Conventional macro-morphology and microscopic examination does not aid in critically distinguishing the three species. DNA markers were developed by amplifying and sequencing the complete internal transcribed spacer region (ITS1, 5.8S rRNA and ITS2) from the genomic DNA, using universal primers. The markers developed are efficient and reliable in authenticating B. aristata, B. asiatica and B. lycium. These are useful as molecular pharmacognostic tool in quality control of raw drugs.
doi:10.1007/s13205-010-0001-5
PMCID: PMC3339604  PMID: 22558531
Daruharidra; Berberis spp.; Quality control; Internal transcribed spacer (ITS); DNA marker; Chemistry; Biotechnology; Stem Cells; Biomaterials; Bioinformatics; Agriculture; Cancer Research

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