Annals of Phytomedicine; An International Journal

3

Authors: Md. Liyakat Ahmed

The research to discover a new drug is never ending phenomenon. The main aim of experimental pharmacology is to find out a therapeutic agent, suitable for human use. Preclinical experimental pharmacology involves the identification and optimization of novel chemical lead structures and testing on animals and animal tissues or organs for their biological actions. A technique designed for pharmacy and non-pharmacy students in particular for bioassay method and in situ method, to observe the effect of various drugs in isolated organs of the animal in experimental pharmacology. Recording levers are used to record the contraction or relaxation of the isolated tissue preparation. A celluloid tip of refill cut from marketed sketch- pen and pointer is fixed to it, which can be used along with simple, frontal writing and starling’s heart levers etc . This new technique is designed and practiced by Md. Liyakat Ahmed , since 2007. Hence, it is named as ‘Liy-levers’ technique. The parasympathetic effect of acetylcholine on frog’s rectus abdominis, isolated muscle preparation by using interpolation bioassay method and effect of various drugs on isolated frog’s heart, results shown in the form of graphical peaks, recorded by the following same technique

4

Authors: Rajeswari Satapathy and Paramjyoti Swamy

Herbal drugs are traditionally used in various parts of world to cure different diseases. In the present research studies, Peltophorum pterocarpum ( DC ) K Heyne was used as herbal drug and its hepatoprotactive effect in CCl 4 induced liver injury in mice was evaluated, using essential biochemical parameters like total bilirubin, SGOT , SGPT and ALP . In this experiment, pretreatment of mice with Peltophorum pterocarpum leaf extracts, resulted in significant reduction in all the liver marker enzymes and bilirubin. Total phenolic and flavonoid contents were evaluated according to Folin-Ciocalteau procedure and a calorimetric method, respectively. It showed high content of total phenolic compounds, total flavonoids and polyphenolic compounds.

5

Authors: Kuldeep Singh Yadav, Narayan P. Yadav, Naveen Kumar and Suaib Luqman

Leucas cephalotes Spreng (Family: Lamiaceae) known as ‘Dronapushpi’ in Sanskrit and ‘Guma’ in Hindi, is being extensively used by rural people of North India to treat various ailments including inflammation. Therefore, present study was undertaken to validate the effect of L. cephalotes extracts in topical inflammation, using mouse ear edema model, induced by 12-O-tetradecanoylphorbol-13-acetate ( TPA ). The anti-inflammatory effect was evaluated by measuring ear weight, level of tumor necrosis factor ( TNF )- α , interleukin ( IL )- 1 β , interleukin ( IL )- 6 , activity of lipid peroxidation ( LPO ) and the production of nitric oxide ( NO ). The results showed that non-polar fraction of methanolic extract ( MNF ) of L. cephalotes significantly decreased the ear weight, LPO activity and NO level. Additionally, MNF remarkably inhibited the expression of TNF- α , IL-1 β and IL-6 while polar fraction of methanolic extract ( MPF ) considerably reduced LPO activity and IL-6 level only. The results indicate that non-polar fraction of methanolic extract ( MNF ) of L. cephalotes has potential anti-inflammatory effect on TPA -induced inflammation in mice ear

6

Authors: Syed Mohammed Kazim and Mohammed Ibrahim

The study is based upon the formulation of solid dosage forms, using natural ingredients and a novel method development for the estimation of individual ingredients in various combination units. Tablets of analgesic drug mafenemic acid in combination with dicyclomine, a smooth muscle relaxant was prepared. A hepatoprotective agent DL -methionine was also incorporated in the formulation. Enhancement of bioavailability of analgesic tablets by introducing the seeds of Lallemantia royleana Benth (commonly known as Balanga) was achieved, accompanied with the estimation of the amount of methionine, incorporated in the formulation. The mathematical tool of orthogonal polynomials was implemented for the above estimation of percentage purity of a particular ingredient in the above complex formulation.The novel formulation is evaluated in terms of its bioavailability by comparing the parameters like disintegration time and dissolution time, with that of the standard formulations. The above assessment was carried out to establish the role of the natural disintegrant used. There are many methods to enhance bioavailability; one of the methods is the reduction of disintegration time. Balanga seeds are used for the above purpose as the seeds are known to contain plenty of mucilage within its chemical constituents. The mucilage absorbs water by imbibition and helps in tablet disintegration

7

Authors: Sakina Razack and Farhath Khanum

Anxiety partly caused by stress has emerged to be a common psychiatric manifestation of the modern world and lifestyles. Medications for anxiety viz ., benzodiazepines, MAOI’s have limitations due to unbearable side effects. Herbal anxiolytes promise to alleviate anxiety and other pscychiatric disorders with minimal adverse effects. Nardostachys jatamansi DC belongs to plant family Valerianaceae and a prominent herb used in traditional medicine for a wide range of disorders. There is limited scientific literature with regard to its anxiolytic activity. The purpose of the present study was to investigate the anxiolytic effects of water (cold and hot) and 70% ethanolic extract of the rhizome of Nardostachys jatamansi . The effect on anxiety was assessed using Elevated plus maze and Open field test. Pre-treatment of mice with different doses of the extract (125, 250 and 500 mg/ kg body weight) significantly (p<0.01) alleviated anxiety by increasing the number of entries on the open arm, time spent on open arms, percent time spent on open arms and decreasing the time spent on closed arms in the elevated plus maze test. Moreover, the 70% ethanolic extract also significantly (p<0.01) increased the locomotor activity as observed in the open field test confirming the anxiolytic effects in mice. Diazepam was used as a standard to compare the effects. Correlation analysis showed a linear relationship between antioxidant potency and radical scavenging activity with the polyphenolic content

8

Authors: vinita Puranik, D.K. Chauhan, Vandana Mishra and G.K. Rai

Many countries are registering their standardized herbal drugs with proven clinical post-harvest preservation and management methods are numerous, but many are not economically feasible. Perhaps one of the most widely used herb preservation methods is dehydration. Care must be taken to set drying parameters such that the herbs are not exposed to excessively high temperatures, causing major losses of their medicinal, culinary, visual and nutraceutical properties, negatively affecting the product value. The leaves of Boerhaavia diffusa L. and Andrographis paniculata (Burm.f.) Wall. ex Nees are used for the treatment of many diseases but there was no report about the effects of different drying methods on physicochemical quality and bioactive compounds in leaves of A.paniculata and B.diffusa . Therefore, different drying methods were applied to leaves, and the influence of these methods to physicochemical quality, the total phenols and percentage radical scavenging activity was investigated, with the aim to pick out the practical drying method. From the results, it was found that freeze drying produced the best quality leaf powders in terms of ascorbic acid, polyphenolic content and antioxidant activity, though it was quite hygroscopic in nature. It can be inferred that A.paniculata and B.diffusa leaf powders are good source of natural antioxidants which can be an alternative to synthetic antioxidants. The obtained results for leaf powder gave justifications for further investigations on its applicability as a functional food.

8

Authors: Syed Asad Bakhtiary, M. M. Iqbal and Md. Ibrahim

Conventional drugs used in the treatment of liver and kidney diseases are sometimes inadequate and can lead to serious side effects. Therefore, it is necessary to search for alternative drugs for the treatment of liver and kidney diseases in order to replace currently used drugs of doubtful efficacy and safety. Leaves of Phyllanthus amarus Schum & Thonn (Family: Euphorbiaceae) is traditionally known for its liver and kidney protection. We have made an attempt to find out the hepatoprotective and nephroprotective activity on Phyllanthus amarus seeds. Hepato and nephrotoxicity was induced by thioacetamide (100 mg/kg s.c) and gentamycin (80 mg/kg i.p), respectively. Phyllanthus amarus seed extracts (70% methanolic 250 mg/kg and aqueous 300 mg/kg) were administered orally to Wistar rats. Hepatoprotection was assessed by measuring the biochemical parameters like serum glutamate pyruvate transaminase (SGPT) , serum glutamate oxaloacetate transaminase (SGOT) , alkaline phosphatase (ALP) , total bilirubin, direct bilurubin, triglycerides, cholesterol and histopathological changes in liver. Whereas nephroprotection was assessed by measuring serum creatinine, blood urea nitrogen (BUN) and kidney weight. Histopathological analysis of liver samples also confirmed the hepatoprotective activity of methanolic extract of the seeds, which was comparable to the standard silymarin. The results obtained in this study indicate that the methanolic extract has significant effect than aqueous extract when compared to silymarin and cystone, respectively. The mechanism involved in the protection could be associated with its strong capability to reduce the enhanced levels of above parameters. Hence, the Phyllanthus amarus seed possesses a potent protective effect against thioacetamide-induced hepatic damage, and gentamycin-induced renal damage

9

Authors: Mohammed Majid Iqbal, Sanjeev Gupta, Shanti Sagar and Mohammed Ibrahim

This present research study was carried out to develop and evaluate the subcutaneous implants of chitosan for the administration of tramadol. Tramadol is one of the centrally acting analgesics with opioid agonist properties and can be safely used in such condition without the side effects. Rod shaped implants of 1 mm diameter and 2.7 mm in length were prepared by extrusion method, using a specially designed galaxy extruder. Implants were analyzed for drug content uniformity, thickness, weight variation, short term stability study. In vitro release study in phosphate buffer at 37 ± 1 o C, demonstrated that the rate of release of the tramadol from the implant matrix was a function of concentrations of the polymer. The implant formulations, having different concentrations of crosslinking, show varied drug release, can be extended for 17 days, compared to drug released from implants with no crosslinking. Thus, crosslinikng effectively control the amount of drug release. The release of drug from all implant formulations was uniform and was spread over a period of 17 days. The implant formulations were found sterile, uniform in weight, drug content and size. Short term stability studies of drug implants revealed that the implants formulations were stable, and there were no significant changes in the physical appearance and drug content of the implants formulations. Data obtained from the study suggest that the implants prepared from chitosan would be promising as an interesting biodegradable system for sustained delivery of tramadol for the pain management

4

Authors: C. Beena and V.V. Radhakrishnan

S araca asoca ( Roxb.) de Wilde , is locally known as Sita asok and is grown as an ornamental tree throughout the country. But bark of asoka tree is an important herbal raw drug used in gynaecological problems like uterine bleeding associated with fibroids. It is a major constituent of the ayurvedic medicines “asokarishtam” and “asokagritham”used for the treatment of gynaecological disorders. The increasing demand of the bark along with less availability has led to its widespread adulteration. It is widely adulterated with the bark of Polyalthia longifolia, a well known avenue tree available in plenty. Since adulteration of herbal drugs has clinically relevant effects, an attempt has been made to develop an easy method to detect the adulteration in market samples of the raw drug asoka bark and to develop specific chemical fingerprints of the genuine bark by TLC (Thin Layer Chromatography). Petroleum ether extracts of the stem barks of asoka and polyalthia were found best suitable to develop good chromatograms. These specific chromatograms proved efficient in detecting adulteration in 22 market samples analysed

5

Authors: Md. Liyakat Ahmed,Ashwini Jadhav, Paramjyothi Swamy*, Syed Sanaullah and N. Santosh Kumar

The antiulcer effect of Semicarpus anacardium Linn. seed extracts was investigated against aspirin plus pylorus ligation induced and ethanol induced gastric ulcers in rats. Both the models revealed ulcer healing property of the seed extracts. The present investigation also revealed the hepatoprotective activity of the seed extracts, against ethanol induced hepatotoxicity. Histopathological studies of the liver showed significant restoration of the normal histomorphological pattern of hepatocytes. The biochemical estimation of serum bilirubin, serum glutamate pyruvate transaminase ( SGPT ), serum glutamate oxaloacetate transaminase ( SGOT ) and alkaline phosphatase ( ALP ) showed significant reduction in the rats fed with the seed extracts. The study, thus, substantiates the potential anticulcer and hepatoprotective effects of Semicarpus anacardium seeds

6

Authors: Saujanya Kumar Jha and Shailendra Kumar Srivastava

The objective of this study is to determine the in vivo effect of methanolic extracts of leaves and seeds of Trigonella foenum-graecum Linn. in blood plasma of mercuric chloride induced oxidative stress in mice, with respect to the levels of different antioxidant enzyme systems like catalase, Super oxide dismutase (SOD) and glutathione reductase enzyme activities. DPPH assay of the methanolic extracts of the seeds and leaves were conducted and it was seen that the free radical scavenging activity of seed was better than that of methanolic extract of leaves as seen by a lower E.C. 50 value obtained for seed as compared to a higher E.C. 50 value for the leaves. It was seen that there was an enhanced lipid peroxidation and antioxidant enzyme levels were significantly decreased in blood plasma of mercuric chloride treated mice. Subsequent oral administration of methanolic extract of Trigonella foenum-graecum Linn. seeds and leaves for seven days, demonstrated decreased lipid peroxidation in blood plasma of mice almost to the normal control range. Antioxidant enzyme levels post extract feeding also increased in contrast to decrease in their levels on mercuric chloride administration. However, the seeds were found to have a better antioxidant potential than the leaves in vivo

6

Authors: N. Lakshmi, Divya Pingili and Sheena M Raj

This research paper reports a comparision of the genomic DNA constitution between two selected antimalarials plants, i.e ., Eucalyptus globulus Labill. and Artemisia annua L. The Random Amplification of Polymorhic DNA (RAPD) analysis was performed to detect the polymorphism between them. Genetic analysis of these plants relies on high yields of pure DNA samples. Optimized conditions for DNA isolation and Polymerase Chain Reaction (PCR) polymerization were provided. The method involves a modified Cetyltrimethyl Ammonium Bromide (CTAB) extraction employing polyvinyl pyrrolidone. The yield of DNA ranged from 1-2μg/ μl per gram of the leaf tissue and purity (ratio) was between 1.6-1.7, indicating minimal levels of contaminating metabolites. The isolated DNA was used for RAPD analysis. Reproducible and amplifiable products were observed in all PCR reactions that are polymorphic. The analysis revealed that these two plants showed genomic similarities, hence, it may be useful in paving a gateway for further research to design new modified drugs for malaria.

7

Authors: Divya Pingili, Sneha Jijabapu Anarthe, Nulgumnalli Manjunathaiah Raghavendra

In the present investigation, methanolic and hydro alcoholic polyherbal extracts . having Terminalia arjuna (Roxb.) Wight & Arn, Piper nigrum Linn. and Cuminum cyminum Linn. were evaluated for their in vitro antioxidant activity by 1,1-diphenyl- 2-picryl hydrazyl (DPPH) and superoxide radical scavenging method; in vitro antidiabetic activity by α -glucosidase and α -amylase inhibitory method; and anticancer activity by 3-(4,5-dimethylthiazole-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay against A549 human lung cell carcinoma. Antioxidant activity of methanolic extract of Terminalia arjuna (S 1 ) was found to be more potent compared to other polyherbal extractions. S 1 also inhibited α -glucosidase enzyme and hydroalcoholic extract inhibited α -amylase enzyme. Few polyherbal combinations are found to possess significant anticancer activity against A549 human lung cell carcinoma.

8

Authors: N.M.A. Rasheed, K. Nagaiah, P.R. Goud and V.U.M. Sharma

Many countries are registering their standardized herbal drugs with proven clinical efficacy, safety and as dietary supplements. But India is unable to exploit the World market due to unsatisfactory system of quality control which necessitates the establishment of standards for herbal single drugs and compound formulations. In order to ensure international community to use AYUSH drugs with confidence and without compunction, developing acceptable quality control standards of herbal medicines aims to ensure their consistency, safety and efficacy with good manufacturing practices is very much essential. Study of chemical markers is applicable to many research areas, including authentication of genuine species, search for new resources or substitutes of raw materials, optimization of extraction and purification methods, structure elucidation and purity determination. Systematic investigations using chemical markers may lead to discoveries and development of new drugs

8

Authors: Suaib Luqman and Ritesh Kumar

The present study investigates a concentration-dependent assessment of scavenging property and antioxidant activity in the extracts of Emblica officinalis Gaertn. syn. Phyllanthus emblica L. fruit using various in vitro assays. A correlation of aqueous and ethanolic extract of Emblica fruit along with different known standards for its antioxidative properties was made by performing assays like TPC, RP, TAC, DPPH FRAP and HRSA . Both the aqueous and ethanolic extract of fruit showed high phenolic content beside strong reducing power and free radical scavenging capacity. The hydroxyl radical scavenging potential of aqueous extract was higher compared to ethanolic extract. A high positive correlation was observed among the in vitro assays for the antioxidative properties except for HRS assay which showed negative corelation. Our finding not only tots up the results related to antioxidant asset of amla fruit but also emphasizes the fact of its usefulness as chemopreventive and/or chemotherapeutic importance. Further studies are warranted to explore the constituents responsible for the antioxidant property and other pharmacological activities

9

Authors: K. Vijaya Sri, D. Srinivas, Ch. Ajay Kumar and D. Ravishanker

The aim of the present study was to evaluate seed flour of Vigna mungo (L.) Hepper or Black gram as binding, disintegration nature and sustained release matrix forming material in tablet formulation. The seeds of Vigna mungo swell and form gelatinous mass when it comes in contact with water due to its hydrophilic nature. Indomethacin ( NSAIDS ) was selected as a model drug because of its poor water solubility and short half life. FT -IR studies showed no evidence on interaction between indomethacin and Vigna mungo . Conventional /sustained release matrix tablets of indomethacin, was developed by using wet granulation technique. Compressed tablets were evaluated for pharmacopoeias tests and compared with marketed product. The kinetic studies showed that the release of drug from formulation CVF2 follows first order model and formulation SVF1 follows zero order model and anomalous diffusion and explained by Higuchi’s model. Selected conventional formulation CVF2 was carried out by anti-inflammatory activity by using plethysmometer. The selected formulations CVF2 and SVF1 was subjected to stability studies for three months at 45ºC temperature with RH 75±5% , and showed stability with respect to release pattern

11

Authors: S. D. Sonkawade and G. R. Naik

Dietary nucleosides and nucleotides perform a range of functions in our body. They play important role in the maintenance of mitochondrial function, differentiation of hematopoietic cells, strengthen the immune system, helps in small intestine growth and development, lipid metabolism etc . External supplementation of dietary nucleotides is a good regimen against mitochondrial dysfunction caused by Nucleosides Reverse Transcriptase Inhibiters ( NRTI ) drug therapy; it has a long history of use for cancer and viral diseases e.g. , NucleomaxX is a dietary nucleotide supplement, and has a potential use in the treatment of mitochondrial toxicity as it contains Mitocnol; a sugarcane extract with a high percentage of nucleotides. Therefore, analysis of these compounds in food materials is very important for improving and assuring food quality as well as to boost Biotech Industrial sector of India; manufacturing a dietary nucleotide supplement. During the present investigation, the extraction protocol for nucleotides from different sources (sugarcane, pulses/lentils, oilseeds, yeast, mushroom and animal milks) was standardized, which is one of the simple, cost effective method for isolation of total nucleotides/nucleosides. The isolated nucleic acid was quantified using double beam UV spectrophotometer. Estimation of RNA was carried out by Orcinol method and DNA by Diphenylamine method, using nucleic acid extract obtained from different food materials. The screening of samples from different sources helped to determine the desirable concentration of nucleic acid in natural sources for dietary nucleotides. Further, investigation is in progress to study influence on animal model, developing therapeutic formulation and scale up criteria