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Ancient Science of Life  1984;3(4):203-206.
Kutaja bija, Kudasappalai or Inderjou is an important seed drug in Ayurveda, Siddha and Unani Medicines. The market sample of Madras Crude drug trade has been identified in our laboratory as the seeds of Holarrhena – anti – dysenterica wall of the family Apocynaceae. The morphology, anatomy, fluorescence analysis and chemical studies of the drugs are reported.
PMCID: PMC3331573  PMID: 22557406
2.  Screening of Antibacterial Potentials of Some Medicinal Plants from Melghat Forest in India 
Cyperus rotundus, Caesalpinia bonducella, Tinospora cordifolia, Gardenia gummifera, Ailanthus excelsa, Acacia arabica, Embelia ribes and Ventilago maderspatana from Melghat forest were screened for their antibacterial potential against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Proteus vulgaris, Salmonella typhi, Shigella flexneri, Salmonella paratyphi, Salmonella typhimurium, Pseudomonas aeruginosa, Enterobacter aerogenes by disc diffusion method. Out of these medicinal plants Caesalpinia bonducella, Gardenia gummifera and Acacia arabica showed remarkable antibacterial potential. The phytochemical analysis had showed the presence of Cardiac glycosides in all extracts (aqueous, acetone, ethanol and methanol) of Acacia arabica, Gardenia gummifera and ethanol, methanol extracts of Caesalpinia bonducella. Flavonoids were present in Gardenia gummifera, Ailanthus excelsa and acetone, methanol extracts of Acacia Arabica. Tannins and phenolic were present in Cyperus rotundus, Embelia ribes, and organic extracts of Ventilago maderspatana.
PMCID: PMC2816464  PMID: 20448847
Antibacterial activity; Melghat; Medicinal Plants; Phytochemical
3.  Standardization of Unani Antidiabetic Tablet - Qurse Tabasheer 
Pharmacognosy Research  2016;8(2):147-152.
Quality control of Unani polyherbal formulations is the need of the day for better acceptance of Unani medicine. Qurse Tabasheer (QT) is a Unani polyherbal formulation containing six ingredients, Tabasheer (Siliceous concretions) (Bambosa arundinaceae Retz.), Gule Surkh (Rosa damascena Mill. flower), Gulnar (Punica granatum Linn. flower), Tukhme kahu (Lactuca sativa Linn. seed), Tukhme khurfa (Portulaca oleraceae Linn. seed), and Gile Armani (bole) widely used in treatment of diabetes. The present study was taken up to scientifically evaluate the various physicochemical parameters to standardize the formulation.
To evaluate various physicochemical parameters including ash values, moisture content, extractive values, thin layer chromatography (TLC) and high-performance TLC (HPTLC), friability, disintegration, uniformity, and weight variation for standardization of QT.
Materials and Methods:
Ingredients were identified by the experts. The method mentioned in national formulary of Unani Medicine with modification was followed for preparation of the tablets. Physicochemical standards were established for ideal batch of tablets on the basis of set parameters regarding friability, hardness, and disintegration. Various parameters such as organoleptic characters, extractive values for the extract and HPTLC fingerprinting postcompression were carried out for evaluation of QT.
Parameters for loss of weight on drying, pH, ash values, extractive values documented. Qualitative chemical tests indicated the presence of alkaloid, glycoside, tannins, and steroids. TLC and HPTLC fingerprinting studies showing the presence of major peaks were documented. Friability, hardness, and disintegration time of ideal batch was 0.09 ± 0.0057, 4.03 ± 0.087, and 25.57 ± 0.4860 min, respectively, and it was found to be within the set limit. Weight variation was <5%. Total fungal and bacterial counts were found to be within the limit.
Standards were established for poly herbal formulation QT, which may be used as reference for preparation and standardization of QT.
In this work Standardization of anti-diabetic tablet Qurse Tabasheer with diverse ingredients including herbal and mineral origin drugs has been attempted with identification of its ingredients, formulation, physicochemical evaluation and HPTLC finger printing, which may help in preparing consistent and better efficacious formulations.
Abbreviations Used: QT: Qurse Tabasheer TLC: thin layer chromatography HPTLC: high-performance thin layer chromatography WHO: World health organization FRLHT: Foundation for Revitalization of Local Health Traditions Fe2O3: Iron oxide Sio2: Silica CaCo3: Calcium carbonate, Tio2: Titanium Oxide NIUM: National Institute of Unani Medicine #: Mesh size LOD: Loss of weight on drying USP: United state Pharmacopeia UV: Ultra Violet λ: Lambda θ: theta CFU: Colony-forming unit
PMCID: PMC4780142  PMID: 27034607
Physicochemical; Qurse Tabasheer; standardization; tablet; Unani
4.  Oxidative DNA damage preventive activity and antioxidant potential of plants used in Unani system of medicine 
There is increasing recognition that many of today's diseases are due to the "oxidative stress" that results from an imbalance between the formation and neutralization of reactive molecules such as reactive oxygen species (ROS) and reactive nitrogen species (RNS), which can be removed with antioxidants. The main objective of the present study was to evaluate the antioxidant activity of plants routinely used in the Unani system of medicine. Several plants were screened for radical scavenging activity, and the ten that showed promising results were selected for further evaluation.
Methanol (50%) extracts were prepared from ten Unani plants, namely Cleome icosandra, Rosa damascena, Cyperus scariosus, Gardenia gummifera, Abies pindrow, Valeriana wallichii, Holarrhena antidysenterica, Anacyclus pyrethrum, Asphodelus tenuifolius and Cyperus scariosus, and were used to determine their total phenolic, flavonoid and ascorbic acid contents, in vitro scavenging of DPPH·, ABTS·+, NO, ·OH, O2.- and ONOO-, and capacity to prevent oxidative DNA damage. Cytotoxic activity was also determined against the U937 cell line.
IC50 values for scavenging DPPH·, ABTS·+, NO, ·OH, O2.- and ONOO- were in the ranges 0.007 ± 0.0001 - 2.006 ± 0.002 mg/ml, 2.54 ± 0.04 - 156.94 ± 5.28 μg/ml, 152.23 ± 3.51 - 286.59 ± 3.89 μg/ml, 18.23 ± 0.03 - 50.13 ± 0.04 μg/ml, 28.85 ± 0.23 - 537.87 ± 93 μg/ml and 0.532 ± 0.015 - 3.39 ± 0.032 mg/ml, respectively. The total phenolic, flavonoid and ascorbic acid contents were in the ranges 62.89 ± 0.43 - 166.13 ± 0.56 mg gallic acid equivalent (GAE)/g extract, 38.89 ± 0.52 - 172.23 ± 0.08 mg quercetin equivalent (QEE)/g extract and 0.14 ± 0.09 - 0.98 ± 0.21 mg AA/g extract. The activities of the different plant extracts against oxidative DNA damage were in the range 0.13-1.60 μg/ml. Of the ten selected plant extracts studied here, seven - C. icosandra, R. damascena, C. scariosus, G. gummifera, A. pindrow, V. wallichii and H. antidysenterica - showed moderate antioxidant activity. Finally, potentially significant oxidative DNA damage preventive activity and antioxidant activity were noted in three plant extracts: C. icosandra, R. damascena and C. scariosus. These three plant extracts showed no cytotoxic activity against U937 cells.
The 50% methanolic extracts obtained from different plant parts contained significant amounts of polyphenols with superior antioxidant activity as evidenced by the scavenging of DPPH·, ABTS·+, NO, ·OH, O2.- and ONOO-. C. icosandra, R. damascena and C. scariosus showed significant potential for preventing oxidative DNA damage and radical scavenging activity, and the G. gummifera, A. pindrow, V. wallichii, H. antidysenterica, A. pyrethrum, A. tenuifolius and O. mascula extracts showed moderate activity. The extracts of C. icosandra, R. damascena and C. scariosus showed no cytotoxicity against U937 cells. In conclusion, these routinely used Unani plants, especially C. icosandra, R. damascena and C. scariosus, which are reported to have significant activity against several human ailments, could be exploited as potential sources of natural antioxidants for plant-based pharmaceutical industries.
PMCID: PMC3020177  PMID: 21159207
5.  Pharmacognostical and physicochemical analysis of Tamarindus indica Linn. stem 
Tamarindus indica Linn. fruits (Chincha) are extensively used in culinary preparations in Indian civilization. Its vast medicinal uses are documented in Ayurvedic classics and it can be used singly or as a component of various formulations. Besides fruit, the Kasta (wood) of T. indica L. is also important and used to prepare Kshara (alkaline extract) an Ayurvedic dosage form. Pharmacognostical and physicochemical details of Chincha Kasta are not available in authentic literature including API (Ayurvedic Pharmacopoeia of India). The study is an attempt in this direction. T. indica L. stem with heartwood was selected and morphological, microscopic and physicochemical standardization characters along with TLC finger print, and fluorescence analysis were documented. Transverse section of stem showed important characters such as phelloderm, stone cells layer, fiber groups, calcium oxalate, crystal fibers, and tylosis in heartwood region. Four characteristic spots were observed under UV long wave, in thin layer chromatography with the solvent combination of toluene: ethyl acetate (8:2). The study can help correct identification and standardization of this plant material.
PMCID: PMC3326798  PMID: 22529673
Ayurveda; Chincha; powder microscopy; tamarind; thin layer chromatography
6.  Pharmacognostic Standardization, Physico- and Phytochemical Evaluation of Amaranthus Spinosus Linn. Root 
Amaranthus spinosus Linn. (Amaranthaceae) is found throughout India. This tree species has been of interest to researchers because it is a medicinal plant employed in the Indian traditional system of medicine. Pharmacognostic standardization; physico-and phytochemical evaluation of the roots of Amaranthus spinosus was carried out, to determine its macro-and microscopical characters, and also some of its quantitative standards. Microscopical studies were done by using the trinocular microscope. Total ash, water-soluble ash, acid-insoluble ash, sulfated ash values, and alcohol-and water-soluble extractive values were determined for physico-chemical evaluations. A preliminary phytochemical screening was also done to detect different phytoconstituents. Microscopically, the root showed cork, cortex, stellar region, and calcium oxalate crystals. Powder microscopy showed anamalous secondary growth in between the xylem vessels and Calcium Oxalate crystals in the cortex region. Total ash was approximately three times more than acid insoluble and water soluble ash. The ethanol soluble extractive was approximately the same as the water soluble extractive. Thin Layer Chromatography (TLC) of the Petroleum-ether extract using Benzene : Ethyl acetate (6 : 1), showed six spots. In the chloroform extract, using Benzene : Ethyl acetate (4 : 1) nine spots were seen, and in the ethanol extract, using Chloroform: Methanol (93 : 7), only four spots were observed, using Iodine vapor as a viewing medium. Phytochemically, the root exhibited terpenes, alkaloids, glycosides, and sugars. These findings might be useful to supplement information with regard to its identification parameters, which are assumed significant in the way of acceptability of herbal drugs, in the present scenario, which lacks regulatory laws to control the quality of herbal drugs.
PMCID: PMC3159276  PMID: 21897662
Amaranthus spinosus Linn.; pharmacognostic standardization; physicochemical evaluations
7.  Standardization of the finished product: Habbe Irqun Nisa - A Unani anti-inflammatory formulation 
Ancient Science of Life  2012;32(1):38-44.
Habb (Pill) is one of the important dosage forms of Unani system of medicine. A number of effective formulations are manufactured in form of Habb because of its various advantages. Out of these, Habbe Irqun Nisa (HI) is a popular anti-inflammatory formulation used in the treatment of Warame Mafasil (arthritis) and Irqun Nisa (sciatica). Nowadays, with increased incidence of these diseases many non-steroidal anti-inflammatory drugs (NSAIDs) are being used in their treatment. Owing to the adverse effects of these drugs, the use of herbal medicines is seen as a better alternative. The basic requirement for the development of Unani system of Medicine is the standardization of single and compound drugs. HI is mentioned in National Formulary of Unani Medicne and selected for the present study.
Materials and Methods:
HI was prepared manually with the powder of crude drugs, passed through sieve no. 100 and mixed with 1% w/w of gum acacia in mucilage form. It was then dried at 60°C for 90 min and then tested for its standardization on different physicochemical parameters, e.g. organoleptic properties, pH values, moisture content, ash values, friability, hardness, weight variation, disintegration time, and thin layer chromatography (TLC).
Results and Conclusion:
The data evolved from this study will make it a validated product and will help in the quality control of other finished products in future research.
PMCID: PMC3733206  PMID: 23929993
Anti-inflammatory; Habb; standardization; Unani system of medicine
8.  Design, formulation and evaluation of caffeine chewing gum 
Caffeine which exists in drinks such as coffee as well as in drug dosage forms in the global market is among the materials that increase alertness and decrease fatigue. Compared to other forms of caffeine, caffeine gum can create faster and more prominent effects. In this study, the main goal is to design a new formulation of caffeine gum with desirable taste and assess its physicochemical properties.
Materials and Methods:
Caffeine gum was prepared by softening of gum bases and then mixing with other formulation ingredients. To decrease the bitterness of caffeine, sugar, aspartame, liquid glucose, sorbitol, manitol, xylitol, and various flavors were used. Caffeine release from gum base was investigated by mechanical chewing set. Content uniformity test was also performed on the gums. The gums were evaluated in terms of organoleptic properties by the Latin-Square design at different stages.
After making 22 formulations of caffeine gums, F11 from 20 mg caffeine gums and F22 from 50 mg caffeine gums were chosen as the best formulation in organoleptic properties. Both types of gum released about 90% of their own drug content after 30 min. Drug content of 20 and 50 mg caffeine gum was about 18.2-21.3 mg and 45.7-53.6 mg respectively.
In this study, 20 and 50 mg caffeine gums with suitable and desirable properties (i.e., good taste and satisfactory release) were formulated. The best flavor for caffeine gum was cinnamon. Both kinds of 20 and 50 mg gums succeeded in content uniformity test.
PMCID: PMC3814650  PMID: 24223387
Caffeine chewing gum; coffee; medicated gum; oral mucosal drug delivery; tea
9.  Antibacterial Activities and In Vitro Anti-Inflammatory (Membrane Stability) Properties of Methanolic Extracts of Gardenia coronaria Leaves 
This work is carried out with Gardenia coronaria leaves that belong to the family Rubiaceae, which is a small-to-medium-sized but tall, deciduous tree, 7.6–9 m high on an average. Leaves are used for the treatment of rheumatic pain and bronchitis. The leaf of the plant consists of coronalolide, coronalolic acid, coronalolide methyl ester, ethyl coronalolate acetate triterpenes (secocycloartanes), and so forth. Methanol extract from the leaves of Gardenia coronaria was completely screened for membrane stability and antibacterial activity. The lower concentrations of Methanolic leaf extract of Gardenia coronaria gave good antimicrobial and anti-inflammatory activity, but higher concentrations gave relatively more projecting antibacterial activity in vitro as compared with Kanamycin. The crude drug's anti-inflammatory effects were compared with those of Aspirin as positive control. The Methanolic extracts of Gardenia coronaria leaves possessed a broad spectrum antibacterial activity against a variety of both Gram-negative and Gram-positive organisms like Streptococcus agalactiae, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, Shigella sonnei, Shigella boydii, and Proteus mirabilis, with a zone of inhibition from 10 to 16 mm. The extract also showed good membrane stability to be considered as having significant anti-inflammatory action.
PMCID: PMC3948643  PMID: 24695677
10.  Reference Genes for qPCR Analysis in Resin-Tapped Adult Slash Pine As a Tool to Address the Molecular Basis of Commercial Resinosis 
Pine oleoresin is a major source of terpenes, consisting of turpentine (mono- and sesquiterpenes) and rosin (diterpenes) fractions. Higher oleoresin yields are of economic interest, since oleoresin derivatives make up a valuable source of materials for chemical industries. Oleoresin can be extracted from living trees, often by the bark streak method, in which bark removal is done periodically, followed by application of stimulant paste containing sulfuric acid and other chemicals on the freshly wounded exposed surface. To better understand the molecular basis of chemically-stimulated and wound induced oleoresin production, we evaluated the stability of 11 putative reference genes for the purpose of normalization in studying Pinus elliottii gene expression during oleoresinosis. Samples for RNA extraction were collected from field-grown adult trees under tapping operations using stimulant pastes with different compositions and at various time points after paste application. Statistical methods established by geNorm, NormFinder, and BestKeeper softwares were consistent in pointing as adequate reference genes HISTO3 and UBI. To confirm expression stability of the candidate reference genes, expression profiles of putative P. elliottii orthologs of resin biosynthesis-related genes encoding Pinus contorta β-pinene synthase [PcTPS-(−)β-pin1], P. contorta levopimaradiene/abietadiene synthase (PcLAS1), Pinus taeda α-pinene synthase [PtTPS-(+)αpin], and P. taeda α-farnesene synthase (PtαFS) were examined following stimulant paste application. Increased oleoresin yields observed in stimulated treatments using phytohormone-based pastes were consistent with higher expression of pinene synthases. Overall, the expression of all genes examined matched the expected profiles of oleoresin-related transcript changes reported for previously examined conifers.
PMCID: PMC4909774  PMID: 27379135
resin; Pinus; gene expression; normalizer genes; terpene synthase
11.  Rheological Characterization and Drug Release Studies of Gum Exudates of Terminalia catappa Linn 
AAPS PharmSciTech  2008;9(3):885-890.
The present study was undertaken to evaluate the gum exudates of Terminalia catappa Linn. (TC gum) as a release retarding excipient in oral controlled drug delivery system. The rheological properties of TC gum were studied and different formulation techniques were used to evaluate the comparative drug release characteristics. The viscosity was found to be dependent on concentration and pH. Temperature up to 60°C did not show significant effect on viscosity. The rheological kinetics evaluated by power law, revealed the shear thinning behavior of the TC gum dispersion in water. Matrix tablets of TC gum were prepared with the model drug dextromethorphan hydrobromide (DH) by direct compression, wet granulation and solid dispersion techniques. The dissolution profiles of the matrix tablets were compared with the pure drug containing capsules using the USP Basket apparatus with 500 ml phosphate buffer of pH 6.8 as a dissolution medium. The drug release from the compressed tablets containing TC gum was comparatively sustained than pure drug containing capsules. Even though all the formulation techniques showed reduction of dissolution rate, aqueous wet granulation showed the maximum sustained release of more than 8 h. The release kinetics estimated by the power law revealed that the drug release mechanism involved in the dextromethorphan matrix is anomalous transport as indicated by the release exponent n values. Thus the study confirmed that the TC gum might be used in the controlled drug delivery system as a release-retarding polymer.
PMCID: PMC2977048  PMID: 18661243
controlled release; dextromethorphan hydrobromide; gum exudates of Terminalia catappa; viscosity
12.  Comparing and authenticating on anatomical aspects of Abrus cantoniensis and Abrus mollis by microscopy 
Pharmacognosy Research  2015;7(2):148-155.
Abrus cantoniensis is popularly used as traditional Chinese medicine and a cool tea in South of China. However, due to diminishing source of A. cantoniensis, it is usually interchanged or adulterated with other species of Abrus genus because of the limited knowledge in identification and differentiation. Especially, Abrus mollis is widely mixed on herbal markets and pharmaceutical preparation.
To ensure safety and efficacy, a detailed comparison was undertaken to carry out an anatomical and micro-morphological study of two species of A. cantoniensis and A. mollis.
Materials and Methods:
Microscopic characteristics of roots, leaves and stems, including transverse sections and the crude drug powder, were observed using a light microscope according to the usual microscopic techniques.
The basic diagnostic features of A. cantoniensis include that stem is extremely thin; xylem vessels of root are radially arranged in 10 or more bundles; pith is hollow in stem, and the palisade tissue is made up of two layers of palisade cells. Furthermore, scanning electron microscopy was used to compare nonglandular hairs and the stomata of the leaflet surface. A table of the key authentication parameters based on the analyzed microscopic characteristics was drawn up.
The study demonstrated that the microscopy and related techniques provided a systematic method that is convenient, feasible, and can be unambiguously applied to the authentication of the species of Abrus.
PMCID: PMC4357965  PMID: 25829788
Abrus cantoniensis; Abrus mollis; authentication; microscopy
13.  Standardization and in vitro antioxidant activity of jatamansi rhizome 
Nardostachys jatamansi Linn. commonly known as jatamansi is a well notorious drug in Indian systems of medicines having various health-related benefits and employed in various herbal formulations due to the presence of high levels of valuable phenolic constituents. The present study was aimed to quality assessment of Jatamansi rhizome by studying macro- and micro-scopic characters along with physicochemical tests, chemo-profiling using thin layer chromatography (TLC), and gas chromatography–mass spectrometry (GC-MS), in vitro antioxidant activity.
Materials and Methods:
Standardization was carried out as per the pharmacopeial guidelines and contaminant estimation was carried out by analyzing the samples for the determination of heavy metals, pesticides, and aflatoxins. Chemo-profiling was done with TLC by optimizing the mobile phase for different extracts. The GC-MS chemo-profiling was also carried out by using hexane soluble fraction of the hydroalcoholic extract. The drug is well known for a protective role in the human body as an antioxidant, so total phenolic contents and in vitro antioxidant efficacy was also determined by using established methods.
The results of quality control and anatomical studies were very much useful for its identification, whereas significant antioxidant efficacy was also observed. The drug was found free of contaminants when analyzed for pesticides and aflatoxins, whereas heavy metals were found under the pharmacopeial limit.
The findings of the present research can be utilized for the identification and quality control of the jatamansi rhizome.
PMCID: PMC4678979  PMID: 26681882
Antioxidant; chemo-profiling; Nardostachys jatamansi Linn.; physicochemical; quality control
14.  Macro-microscopic examination of leaves of Cinnamomum malabatrum (Burm. f.) Blume sold as Tamalapatra 
Ayu  2013;34(2):193-199.
Leaves of Cinnamomum tamala Nees & Eberm. (Lauraceae) commonly known as ‘Tamalapatra’ is a highly reputed commodity in drug and spice trade. Its adulteration with other leaf species belonging to genus Cinnamomum is found to be a common practice in India and other parts of the world. Thorough macroscopic and microscopic investigations are essential to differentiate them. Survey of South Indian crude drug markets revealed that in place of C. tamala some other leaves of Cinnamomum species are sold. Fresh leaves of various Cinnamomum species, including C. tamala, growing in south India were collected and studied to establish their correct identity. Leaves sold in markets of S. India under the name of Tamalapatra were subjected for detailed macro-microscopic evaluation including maceration and powder microscopy. Leaves of Cinnamomum malabatrum showed many distinguishing macro-microscopic characters, which will serve as markers to differentiate them from C. tamala the official source of Tamalapatra. Though macroscopy will serve the purpose of identification of the entire drug, microscopy had revealed the identity of the commercial substitute even in fragmented and powdered form. Macro-microscopic identity of C. malabatrum is established in comparison with the official drug, further chemical and biological studies may be confirmative in deciding the leaves as a substitute or adulterant.
PMCID: PMC3821250  PMID: 24250130
Cinnamomum tamala; Cinnamomum malabatrum; maceration; micrometry; powder microscopy; quantitative microscopy
15.  Effect of Lagenaria siceraria fruit powder on sodium oxalate induced urolithiasis in Wistar rats 
In spite of advances in the present practice of medicine, the formation and growth of calculi continues to trouble mankind, as there is no satisfactory drug to treat kidney stones. In India, many indigenous drugs are in use for the treatment of urinary calculus disease.
The present study was intended to determine anti-urolithiatic effect of Lagenaria siceraria fruit powder (LSFP) against sodium oxalate (NaOx) induced urolithiasis in rats.
Materials and Methods:
Animals were grouped as Vehicle Group (received vehicle gum acacia 2% w/v 1 mL/kg/p.o.), NaOx Group(Sodium oxalate 70 mg/kg,i.p.), LSFP Group (500 mg/kg, p.o. LSFP suspended in gum acacia 2% + Sodium oxalate 70 mg/kg), Cystone Group (500 mg/kg, p.o. Cystone suspended in gum acacia 2% + Sodium oxalate 70 mg/kg).
The increased severity of microscopic calcium oxalate (CaOx) crystals deposition along with increased concentration in the kidney was seen after 7 days of NaOx (70 mg/kg, i.p.) pre-treatment. LSFP (500 mg/kg, p.o.) and standard marketed formulation Cystone (500 mg/kg, p.o.) caused a significant reversal of NaOx-induced changes in ion excretion and urinary CaOx concentration in 7 days treatment.
From the results, it was concluded that LSFP showed beneficial effect against urolithiasis by decreasing CaOx excretion and preventing crystal deposition in the kidney tubules.
PMCID: PMC3371562  PMID: 22707863
Cystone; Lagenaria siceraria; sodium oxalate; urolithiasis
16.  Frankincense essential oil prepared from hydrodistillation of Boswellia sacra gum resins induces human pancreatic cancer cell death in cultures and in a xenograft murine model 
Regardless of the availability of therapeutic options, the overall 5-year survival for patients diagnosed with pancreatic cancer remains less than 5%. Gum resins from Boswellia species, also known as frankincense, have been used as a major ingredient in Ayurvedic and Chinese medicine to treat a variety of health-related conditions. Both frankincense chemical extracts and essential oil prepared from Boswellia species gum resins exhibit anti-neoplastic activity, and have been investigated as potential anti-cancer agents. The goals of this study are to identify optimal condition for preparing frankincense essential oil that possesses potent anti-tumor activity, and to evaluate the activity in both cultured human pancreatic cancer cells and a xenograft mouse cancer model.
Boswellia sacra gum resins were hydrodistilled at 78°C; and essential oil distillate fractions were collected at different durations (Fraction I at 0–2 h, Fraction II at 8–10 h, and Fraction III at 11–12 h). Hydrodistillation of the second half of gum resins was performed at 100°C; and distillate was collected at 11–12 h (Fraction IV). Chemical compositions were identified by gas chromatography–mass spectrometry (GC-MS); and total boswellic acids contents were quantified by high-performance liquid chromatography (HPLC). Frankincense essential oil-modulated pancreatic tumor cell viability and cytotoxicity were determined by colorimetric assays. Levels of apoptotic markers, signaling molecules, and cell cycle regulators expression were characterized by Western blot analysis. A heterotopic (subcutaneous) human pancreatic cancer xenograft nude mouse model was used to evaluate anti-tumor capability of Fraction IV frankincense essential oil in vivo. Frankincense essential oil-induced tumor cytostatic and cytotoxic activities in animals were assessed by immunohistochemistry.
Longer duration and higher temperature hydrodistillation produced more abundant high molecular weight compounds, including boswellic acids, in frankincense essential oil fraactions. Human pancreatic cancer cells were sensitive to Fractions III and IV (containing higher molecular weight compounds) treatment with suppressed cell viability and increased cell death. Essential oil activated the caspase-dependent apoptotic pathway, induced a rapid and transient activation of Akt and Erk1/2, and suppressed levels of cyclin D1 cdk4 expression in cultured pancreatic cancer cells. In addition, Boswellia sacra essential oil Fraction IV exhibited anti-proliferative and pro-apoptotic activities against pancreatic tumors in the heterotopic xenograft mouse model.
All fractions of frankincense essential oil from Boswellia sacra are capable of suppressing viability and inducing apoptosis of a panel of human pancreatic cancer cell lines. Potency of essential oil-suppressed tumor cell viability may be associated with the greater abundance of high molecular weight compounds in Fractions III and IV. Although chemical component(s) responsible for tumor cell cytotoxicity remains undefined, crude essential oil prepared from hydrodistillation of Boswellia sacra gum resins might be a useful alternative therapeutic agent for treating patients with pancreatic adenocarcinoma, an aggressive cancer with poor prognosis.
PMCID: PMC3538159  PMID: 23237355
Apoptosis; Boswellia sacra, Boswellic acid; Essential oil; Frankincense; Hydrodistillation; Pancreatic cancer
Ancient Science of Life  1984;3(3):140-142.
Kattusirakam or Vanajira is an important fruit drug in Siddha and Ayurveda systems of Medicine. The market sample of Madras has been identified in our laboratory as the fruits, commonly known as seeds of Centratherum anthelminticum (Willd) Kuntz. (Syn. Veronia anthelmintica Willd) of the family Compositate. The morphology, anatomy, fluorescence analysis and chemical characters of the drug are dealt with here.
PMCID: PMC3331554  PMID: 22557396
18.  Chromatographic Profiling of Ellagic Acid in Woodfordia fruticosa Flowers and their Gastroprotective Potential in Ethanol-induced Ulcers in Rats 
Pharmacognosy Research  2016;8(Suppl 1):S5-S11.
Woodfordia fruticosa, a plant of Indian origin, is extensively used in folk medicine for the treatment of various ailments.
The aim of the present study was to standardize the flowers of W. fruticosa, Kurz (Lythraceae), an important plant of Indian origin and explore the chemical constituents contributing to its anti-ulcer activity.
Materials and Methods:
High-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) profiling of the three samples of W. fruticosa flowers purchased from three different markets was done using ellagic acid as the biomarker. Two doses of the aqueous extract of the W. fruticosa (AEWF) flowers were evaluated for anti-ulcer activity by ethanol-induced ulcer model in Wistar albino rats. Omeprazole was used as the positive control. The parameters used for the assessment of the anti-ulcer potential were total titratable acidity (TTA), ulcer index, and percentage protection.
The HPTLC and HPLC studies confirmed the presence of ellagic acid in all the three drug samples. The AEWF showed significant reduction in terms of TTA at both doses of 100 mg/kg and 200 mg/kg. The gastroprotection indicated by a lower ulcer index and higher percentage protection was significant for 200 mg/kg dose of AEWF, better than the protection afforded by omeprazole (10 mg/kg).
The chromatographic profiling and the anti-ulcer studies served as an efficient tool in the characterization of ellagic acid as an important biomarker for the flowers of W. fruticosa and a probable contributor to the gastroprotective capacity of the drug. The bioactivity studies further supported the traditional use of W. fruticosa in the treatment of ulcers.
HPTLC & HPLC fingerprinting of W. fruticosa using ellagic acid as a biomarker.Evaluation of W. fruticosa for gastroprotection potential in ethanol induced gastric ulcer in rats model.Aqueous extract of the drug showed better gastroprotection than the standard drug omeprazole at a dose of 200 mg/kg.
PMCID: PMC4821107  PMID: 27114692
Anti-ulcer activity; ellagic acid; ethanol-induced ulcer; high-performance thin layer chromatography; Woodfordia fruticosa
19.  Phytopharmacological evaluation of ethanol extract of Sida cordifolia L. roots 
To investigate the phytochemical screening (group determination) and selected pharmacological activities (antioxidant, antimicrobial and analgesic activity) of the plant Sida cordifolia Linn (S. cordifolia).
Eighty percent concentrated ethanol extract of the roots was used. To identify the chemical constituents of plant extract standard procedures were followed. In phytochemical screening the crude extract was tested for the presence of different chemical groups like reducing sugar, tannins, saponins, steroids, flavonoids, gums, alkaloids and glycosides. The antioxidant property of ethanolic extract of S. cordifolia was assessed by DPPH free radical scavenging activity. Analgesic activity of the extract was tested using the model of acetic acid induced writhing in mice. Diclofenac sodium is used as reference standard drug for the analgesic activity test. Antibacterial activity of plant extract was carried out using disc diffusion method with five pathogenic bacteria comparison with kanamycin as a standard.
Phytochemical analysis of the ethanolic extract of the roots of S. cordifolia indicated the presence of reducing sugar, alkaloids, steroids and saponins. In DPPH scavenging assay the IC50 value was found to be 50 µg/mL which was not comparable to the standard ascorbic acid. The crude extract produced 44.30% inhibition of writhing at the dose of 500 mg/kg body weight which is statistically significant (P>0.001). The in vitro antimicrobial activity of the ethanol extract of the roots of S. cordifolia showed no antimicrobial activity against five types of microorganisms. The experiment was conducted only with five species of bacteria as test species, which do not at all indicate the total inactivity against micro-organisms.
The obtained results provide a support for the use of this plant in traditional medicine but further pharmacological studies are required.
PMCID: PMC3819490  PMID: 24144125
Antioxidant; Antimicrobial; Analgesic; DPPH; Phytochemical screening
20.  A Clinico-analytical Study on Seed of Wrightia antidysenterica Linn. as a Therapeutic Emetic Agent (Vamaka Yoga) in the Management of Psoriasis 
Pharmacognosy Research  2016;8(Suppl 1):S19-S25.
Wrightia antidysenterica Linn. (WA) is male variety Kutaja stated to be potent therapeutic emetic agent in skin disorders. Expulsion of doshas through oral route is termed as Vamana Karma (VK) (therapeutic emesis). However, so far, its utility for Vamana is not explored in detail, therefore there is a need to revalidate the utility of WA for Vamana. Hence, the above study was conducted to ascertain the efficacy as a therapeutic emetic agent (vamaka yoga) in the management of psoriasis along with quality control and standardization of this herb.
Materials and Methods:
The drug was standardized as per analytical procedures in Pharmacopeias. Thirty patients of psoriasis fulfilling inclusion criteria were taken for the study and Vamana with WA was conducted. Criteria were prepared to assess the signs and Symptoms of psoriasis. VK was assessed using the classical Lakshanas (features) such as Anthiki shudhi (Ending symptoms of emesis), Vaigiki shudhi (features of vomiting bouts), Maniki shudhi (Quantitative and qualitative purification), complications.
VK with WA showed significant relief in parameters of psoriasis such as scaling, itching, candle grease sign (P < 0.001), and psoriasis area and severity index score (P = 0.001). In VK with WA, mean number of Vegas (vomiting bouts) was 6.91. 66% patients showing quantitative purification between 301 and 600 ml. 73.33% showed all Symptoms of purification. 73.33% patients showed Kaphanta vamana (Moderate expulsion of desire humor). In the level of biopurification, 66.66% patients showed moderated purification. No complication was noted with moderate drug palatability.
Pharmacopeial analytical study showed its standardized values for testing the drug used for the study. It is proved as potent therapeutic emetic agent with no complication showed its clinical benefits over skin disorder like psoriasis.
Seeds of Wrightia antidysenterica (WA) Linn. free from any foreign matter were selected for the study. Loss on drying revealed 6.535% moisture content; total ash indicating of total inorganic content was found to be 5.12%; acid insoluble ash is the acid insoluble part of total ash, mainly silica, WA showed 0.393% acid insoluble ash; ethanol and water soluble extractive is indicative of percentage active constituents were found to be 25.66 and 20.854%, respectively. High-performance thin layer chromatography fingerprinting profiles of WA under 254 nm showed the presence of 7 spots (all in green) at Rf values ranging from 0.21 to 0.88. Under 366 nm there were 4 prominent spots (all in fluorescent) at Rf 0.49 to 0.82 and, when scanned under white light 620 nm following derivatization with vanillin sulfuric acid 6 spots (in different colors) were evident at Rf 0.28 to 0.58. Among these spot with Rf of 0.58 was common when visualized under all the three methods. Rf values by densitometric scan of WA showed 12 peaks at 254 nm and 5 peaks at 366 nm. However, in clinical trial, it was found to be a potent emetic agent without any complication.
Abbreviations Used: WA: Wrightia antidysenterica; Linn.; VK: Vamana karma; BT: Before treatment; FP: Freidman's P value; CHS: Chi-square value; NR: Negative ranks; PR: Positive ranks; N: Sample number, AS: Austipz sign; CG: Candle grease test; SSL: Samyak Snigdha Lakshana
PMCID: PMC4821102  PMID: 27114687
Kutaja beej; pharmacopeial analysis; psoriasis; therapeutic emesis; vamana
21.  A Collaborative Epidemiological Investigation into the Criminal Fake Artesunate Trade in South East Asia  
PLoS Medicine  2008;5(2):e32.
Since 1998 the serious public health problem in South East Asia of counterfeit artesunate, containing no or subtherapeutic amounts of the active antimalarial ingredient, has led to deaths from untreated malaria, reduced confidence in this vital drug, large economic losses for the legitimate manufacturers, and concerns that artemisinin resistance might be engendered.
Methods and Findings
With evidence of a deteriorating situation, a group of police, criminal analysts, chemists, palynologists, and health workers collaborated to determine the source of these counterfeits under the auspices of the International Criminal Police Organization (INTERPOL) and the Western Pacific World Health Organization Regional Office. A total of 391 samples of genuine and counterfeit artesunate collected in Vietnam (75), Cambodia (48), Lao PDR (115), Myanmar (Burma) (137) and the Thai/Myanmar border (16), were available for analysis. Sixteen different fake hologram types were identified. High-performance liquid chromatography and/or mass spectrometry confirmed that all specimens thought to be counterfeit (195/391, 49.9%) on the basis of packaging contained no or small quantities of artesunate (up to 12 mg per tablet as opposed to ∼ 50 mg per genuine tablet). Chemical analysis demonstrated a wide diversity of wrong active ingredients, including banned pharmaceuticals, such as metamizole, and safrole, a carcinogen, and raw material for manufacture of methylenedioxymethamphetamine (‘ecstasy'). Evidence from chemical, mineralogical, biological, and packaging analysis suggested that at least some of the counterfeits were manufactured in southeast People's Republic of China. This evidence prompted the Chinese Government to act quickly against the criminal traders with arrests and seizures.
An international multi-disciplinary group obtained evidence that some of the counterfeit artesunate was manufactured in China, and this prompted a criminal investigation. International cross-disciplinary collaborations may be appropriate in the investigation of other serious counterfeit medicine public health problems elsewhere, but strengthening of international collaborations and forensic and drug regulatory authority capacity will be required.
Paul Newton and colleagues' international, collaborative study found evidence that counterfeit artesunate was being manufactured in China, which prompted a criminal investigation.
Editors' Summary
Malaria is one of the world's largest public health problems, causing around 500 million cases of illness and at least one million deaths per year (the estimates vary widely). The most serious form of malaria is caused by the parasite Plasmodium falciparum, which has become resistant to multiple drugs that had previously been the cornerstones of antimalarial regimens. One group of drugs for treating malaria, the artemisinin therapies including artesunate, are based upon a Chinese herb called qinghaosu; these have now become vital to the treatment of P. falciparum malaria. But counterfeit artesunate, containing none or too little (“subtherapeutic levels”) of the active ingredient, is a growing problem especially in South and East Asia. Fake artesunate is devastating for malaria control: it causes avoidable death, reduces confidence in the drug, and takes away profit from legitimate manufacturers. Of major concern also is the potential for subtherapeutic counterfeit artesunate to fuel the parasite's resistance to the artemisinin group of drugs.
Previous estimates have suggested that between 33% and 53% of artesunate tablets in mainland South East Asia are counterfeit. In this paper the authors report on an unprecedented international collaboration and criminal investigation that attempted to quantify and source counterfeit artesunate among some of the most malarious countries in Asia.
Why Was This Study Done?
Previous reports have identified the problem of fake artesunate, but as of yet there have been few reports on the potential solutions. Concerned health workers and scientists, the regional World Health Organization (WHO) office and the International Criminal Police Organization (INTERPOL) got together to discuss what could be done in May 2005 when it became clear the counterfeit artesunate situation was worsening in the Greater Mekong Sub-Region of South East Asia (comprising Cambodia, Lao People's Democratic Republic, Myanmar, Thailand, Vietnam, and Yunnan Province in the People's Republic of China). Their subsequent investigation combined the goals and methods of a range of concerned parties—police, scientists, and health workers—to identify the source of counterfeit artesunate in South East Asia and to supply the evidence to help arrest and prosecute the perpetrators.
What Did the Researchers Do and Find?
The researchers conducted forensic analyses of samples of genuine and counterfeit artesunate. They selected these samples from larger surveys and investigations that had been conducted in the region beginning in the year 2000. Genuine samples were supplied by a manufacturer to provide a comparator. The authors examined the physical appearance of the packages and subjected the tablets to a wide range of chemical and biological tests that allowed an analysis of the components contained in the tablets.
When comparing the collected packages and tablets against the genuine samples, the researchers found considerable diversity of fake artesunate in SE Asia. Sixteen different fake hologram types (the stickers contained on packages meant to identify them as genuine) were found. Chemical analysis revealed that all tablets thought to be fake contained no or very small quantities of artesunate. Other ingredients found in the artesunate counterfeit tablets included paracetamol, antibiotics, older antimalarial drugs, and a range of minerals, and there were a variety of gases surrounding the tablets inside the packaging. Biological analyses of pollen grains inside the packaging suggested that the packages originated in the parts of South East Asia along the Chinese border.
What Do these Findings Mean?
The results were crucial in helping the authorities establish the origin of the fake artesunate. For example, the authors identified two regional clusters where the counterfeit tablets appeared to be coming from, thus flagging a potential manufacturing site or distribution network. The presence of wrong active pharmaceutical ingredients (such as the older antimalarial drugs) suggested the counterfeiters had access to a variety of active pharmaceutical ingredients. The presence of safrole, a precursor to the illicit drug ecstasy, suggested the counterfeits may be coming from factories that manufacture ecstasy. And the identification of minerals indigenous to certain regions also helped identify the counterfeits' origin. The researchers concluded that at least some of the counterfeit artesunate was coming from southern China. The Secretary General of INTERPOL presented the findings to the Chinese government, which then carried out a criminal investigation and arrested individuals alleged to have produced and distributed the counterfeit artesunate.
The collaboration between police, public health workers and scientists on combating fake artesunate is unique, and provides a model for others to follow. However, the authors note that substantial capacity in forensic analysis and the infrastructure to support collaborations between these different disciplines are needed.
Additional Information.
Please access these Web sites via the online version of this summary at
The World Health Organization in 2006 created IMPACT—International Medical Products Anti-Counterfeiting Taskforce—with the aim of forging international collaboration to seek global solutions to this global challenge and in raising awareness of the dangers of counterfeit medical products. The task force membership includes international organizations, nongovernmental organizations, enforcement agencies, pharmaceutical manufacturers' associations, and drug and regulatory authorities. IMPACT's Web site notes that trade in counterfeit medicines is widespread and affects both developed and developing countries but is more prevalent in countries that have weak drug regulatory systems, poor supply of basic medicines, unregulated markets, high drug prices and/or significant price differentials. IMPACT holds international conferences and maintains a rapid alert system for counterfeit drugs.
The drug industry's anticounterfeit organization, Pharmaceutical Security Institute, works to develop improved systems to identify the extent of the counterfeiting problem and to assist in coordinating international inquiries. Its membership includes 21 large pharmaceutical companies.
The Web site of David Pizzanelli, a world expert on security holography, contains a PowerPoint presentation co-authored by Paul Newton that illustrates the different types of fake holograms found on fake artesunate packages, and their implications for artemisinin resistance (
PMCID: PMC2235893  PMID: 18271620
22.  Pharmacognostical studies of leaves of Combretum albidum G. Don 
Ancient Science of Life  2013;32(4):187-192.
Combretum albidum Don belonging to family Combretaceae is an unexplored medicinal plant in the Indian medicinal system. According to ethnobotanical information, the leaves are used in the treatment of peptic ulcer and its fruits are used in diarrhoea and dysentery. Stem bark is used in the treatment of jaundice and skin diseases. The problem encountered in standardisation of this medicinal plant is its identification by source.
Materials and Methods:
The pharmacognostical studies were carried out in terms of organoleptic, macroscopic, microscopic, physicochemical, florescence and phytochemical analysis. Physicochemical parameters such as total ash, moisture content and extractive values are determined by World Health Organization guidelines. The microscopic features of leaf components are observed with Nikon lab photo device with microscopic units.
Macroscopically, the leaves are simple, obovate in shape, acuminate apex, entire margin and smooth surface. Microscopically, the leaves showed a large vascular strand that consists of thick walled xylem elements mixed with xylem fibres and phloem which is present in a thin layer along inner and outer portions of xylem. External to the xylem occur a thin line of sclerenchyma. Powder microscopy revealed glandular trichomes in the adaxial epidermal peelings also shows the non-glandular trichomes fairly common in powder and epidermis with anisocytic stomata. Vessels elements are narrow, long, cylindrical and dense multi-seriate bordered pits. Xylem fibres are thin and long, with thick walls, which are lignified. Preliminary phytochemical screening showed the presence of carbohydrate, glycoside, saponin, flavonoid, phytosterols and phenolic compounds.
The results of the study can serve as a valuable source of pharmacognostic information as suitable standards for identification of this plant material in future investigations and applications.
PMCID: PMC4078467  PMID: 24991065
Combretum albidum; fluorescence analysis; macroscopy; microscopy; physicochemical; phytochemical
Ancient Science of Life  2002;22(1):67-75.
The plant Coldenia procumbens Linn. is used commonly in Indian system of medicine for various ailments. The present paper deals with detailed pharmacognosy of the leaf of coldenia procumbens Linn. and includes its Macro/Micro morphological (vein islet, vein termination numbers and stomatal index) anatomical characters, Physico chemical standards such as ash values, extractive values, crude fibre content and fluorescence characters of various extracts and leaf powder after treatment with different chemical reagents under UV light. Prelimanary phytochemical tests on various extracts of the leaf have also been carried out.
PMCID: PMC3330985  PMID: 22557078
24.  Ancient-modern concordance in Ayurvedic plants: some examples. 
Environmental Health Perspectives  1999;107(10):783-789.
Ayurveda is the ancient (before 2500 b.c.) Indian system of health care and longevity. It involves a holistic view of man, his health, and illness. Ayurvedic treatment of a disease consists of salubrious use of drugs, diets, and certain practices. Medicinal preparations are invariably complex mixtures, based mostly on plant products. Around 1,250 plants are currently used in various Ayurvedic preparations. Many Indian medicinal plants have come under scientific scrutiny since the middle of the nineteenth century, although in a sporadic fashion. The first significant contribution from Ayurvedic materia medica came with the isolation of the hypertensive alkaloid from the sarpagandha plant (Rouwolfia serpentina), valued in Ayurveda for the treatment of hypertension, insomnia, and insanity. This was the first important ancient-modern concordance in Ayurvedic plants. With the gradual coming of age of chemistry and biology, disciplines central to the study of biologic activities of natural products, many Ayurvedic plants have been reinvestigated. Our work on Commiphora wightti gum-resin, valued in Ayurveda for correcting lipid disorders, has been described in some detail; based on these investigations, a modern antihyperlipoproteinemic drug is on the market in India and some other countries. There has also been concordance for a few other Ayurvedic crude drugs such as Asparagus racemosus, Cedrus deodara, and Psoralea corylifolia.
PMCID: PMC1566595  PMID: 10504143
25.  Anti-Streptococcal activity of Brazilian Amazon Rain Forest plant extracts presents potential for preventive strategies against dental caries 
Caries is a global public health problem, whose control requires the introduction of low-cost treatments, such as strong prevention strategies, minimally invasive techniques and chemical prevention agents. Nature plays an important role as a source of new antibacterial substances that can be used in the prevention of caries, and Brazil is the richest country in terms of biodiversity.
In this study, the disk diffusion method (DDM) was used to screen over 2,000 Brazilian Amazon plant extracts against Streptococcus mutans.
Material and Methods
Seventeen active plant extracts were identified and fractionated. Extracts and their fractions, obtained by liquid-liquid partition, were tested in the DDM assay and in the microdilution broth assay (MBA) to determine their minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs). The extracts were also subjected to antioxidant analysis by thin layer chromatography.
EB271, obtained from Casearia spruceana, showed significant activity against the bacterium in the DDM assay (20.67±0.52 mm), as did EB1129, obtained from Psychotria sp. (Rubiaceae) (15.04±2.29 mm). EB1493, obtained from Ipomoea alba, was the only extract to show strong activity against Streptococcus mutans (0.08 mg/mL
The active extracts, discovered in the Amazon rain forest, show potential as sources of new antibacterial agents for use as chemical coadjuvants in prevention strategies to treat caries.
PMCID: PMC3956399  PMID: 24676578
Streptococcus mutans; Amazonian ecosystem; Plant extracts; Antioxidants; Anti-infective agents

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