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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.  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
3.  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
4.  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
5.  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
6.  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
7.  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
8.  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
9.  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
10.  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
11.  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
12.  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
13.  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
14.  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
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
16.  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
17.  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
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
19.  Pharmacognostic studies of insect gall of Quercus infectoria Olivier (Fagaceae) 
To study the detailed pharmacognostic profile of galls of Quercus infectoria Olivier (Q. infectoria olivier) (Fagaceae), an important medicinal plant used in the Indian system of medicine.
Samples of galls of Q. infectoria were studied by macroscopical, microscopical, physiochemical, phytochemical, fluorescence analysis and othjer methods for standardization as recommended by WHO.
Macroscopically, the crude drug is globose with horny appearances on external surface (1.4-2.3 cm in length and 1-1.5 cm in diameter), with greyish-brown to brownish-black in colour externally and dark brown buff colored. Surface is smooth with numerous horny protuberances giving rough touch, and with unpleasant odour. Microscopically, a wide zone of radially elongated parenchyma cells between upper and lower epidermis were found. The vascular strands were present at all places and radially elongated sclerides touched the lower epidermis. In physico-chemical studies, the moisture, total ash, acid insoluble ash, alcohol soluble, water soluble, petroleum ether, chloroform extractive value and tannin content were found to be 2.790, 5.020, 0.110, 38.780, 41.210, 0.402, 1.590 and 49.200 percentage respectively. Preliminary phytochemical screening showed the presence of phenols, flavonoids, steroids, triterpenes, tannins, saponins and alkaloids.
The results of the present study serve as a valuable source of information and provide suitable standards for identification of this medicinally important plant drug material for future investigations and applications.
PMCID: PMC3819493  PMID: 24144128
Quercus infectoria; Gall extracts; Traditional medicine; Pharmacognostic study; Microscopic studies.
20.  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
Ancient Science of Life  2013;33(1):15-21.
Cardiospermum halicacabum Linn (Sapindaceae) is an important medicinal plant in the traditional system of medicine, known as karṇasphoṭa. The root of it is officially included in Ayurvedic Pharmacopoeia for its therapeutic uses such as jvara, kuṣṭha, pāṇḍu, kṣaya and sandhivāta etc. As no detailed analysis of macroscopy, microscopy characters of the plant, except root, have been carried out till date, it was thought worth to carry out the detailed macroscopic and microscopic study of leaves and stem, following standard pharmacognostical procedures.
Materials and Methods:
Pharmacognostic studies of C. halicacabum were carried out, and in this, the macroscopic, microscopic, physicochemical, fluorescence and phytochemical analyses were done. Physicochemical parameters such as total ash, moisture content, extractive values were determined by World Health Organization guidelines. The microscopic features of leaf and stem components were observed.
Macroscopically the leaves are bi-ternate, ovate-lanceolate in shape with dentate margin. Microscopically, leaf shows prominent midrib and thin dorsiventral lamina. The midrib shows the presence of epidermal layers, angular collenchyma, palisade cells and vascular strands comprised of thin walled xylem and thick walled phloem elements. The lamina shows prominent, narrow and cylindrical upper epidermis. The upper epidermal cells are large and contain mucilage, whereas lower epidermis possesses thin, small and elliptical epidermal cells. The mesophyll was differentiated into two zones upper and lower. The upper zones show narrow cylindrical palisade cells and lower zone shows 2-3 layers of loosely arranged spongy parenchyma cells. In the Paradermal section of the lamina we observe anomocytic stomata. The transverse section of stem shows a pentagonal appearance with five short blunt ridges and prominent cuticle. Parenchymatous cells, cortical sclerenchyma, lignified xylem fibers, phloem and pit were also found. In the powder microscopy of whole plant, glandular trichomes, non-glandular trichomes, fragments of lamina, xylem elements, parenchyma cells and fibers are observed. Phytochemical screening reveals that the C. halicacabum extract contains glycosides, carbohydrates, flavonoids, phytosterols, phenolic compounds and saponin.
Various pharmacognostic characters observed in this study help in identification, quality, purity and standardization of C. halicacabum.
PMCID: PMC4140016  PMID: 25161325
Cardiospermum halicacabum; fluorescence analysis; macroscopy; microscopy; physicochemical; phytochemical
Background & objectives:
Mosquitoes transmit serious human health diseases, causing millions of deaths every year. Plants may be sources of alternative mosquito control agents. The present study was carried out to assess the role of larvicidal activities of the crude extracts of four plants viz. Alternanthera sessilis L. (Amaranthaceae), Trema orientalis L. (Cannabaceae), Gardenia carinata Smith. (Rubiaceae) and Ruellia tuberosa L. (Acanthaceae) against Culex quinquefasciatus Say in laboratory bioassay.
Selective concentrations (0.5, 1 and 1.5%) of crude extract of all four plant leaves were tested against Ist to IVth instar larvae of Cx. quinquefasciatus. Log probit analysis (at 95% confidence level) revealed the LC50 values. Preliminary qualitative phytochemical analyses of crude extracts were also done. The lethal concentrations (%) of crude extracts at 24 h against IIIrd instar larvae were also studied on non-target organisms.
In a 72 h bioassay experiment with crude extract, the highest mortality was recorded in 1.5 per cent extract. A. sessilis showed the highest mortality (76.7 %) at 1.5 per cent crude extract against IInd instar larvae having LC50 value of 0.35 per cent, followed by R. tuberosa (LC50 =1.84%), G. carinata (LC50 = 2.11) and T. orientalis (LC50 = 2.95%). The regression equation showed a dose-dependent mortality, as the rate of mortality (Y) was positively correlated with the concentration (X). Phytochemical analysis of the crude extract showed the presence of many bioactive phytochemicals such as steroids, alkaloids, terpenes, saponins, etc. No changes in the swimming behaviour and survivality of non-target organism were noticed at the studied concentrations.
Interpretation & conclusions:
Crude extract of the four selected plants showed larvicidal activity against Cx. quinquefasciatus. The extracts at the studied concentrations did not produce any harmful effect on non-target organisms.
PMCID: PMC4181141  PMID: 25222784
Culex quinquefasciatus; larvicidal activity; non-target organisms; phytochemical analysis
Ancient Science of Life  2009;29(1):26-28.
The oleoresin of the Shorea robusta Gaertn is called as Shala niryasa, Kala, Sarja rasa which has the chemical constituents such as nor-triterpene, dammarenolic acid, asiatic acid, dipterocarpol, triterpenic acid, tannic acid and phenolic content and possesses antibacterial, analgesic and wound healing effect.
The medicinal property of the plant is highly influenced by the the season in which it is cultivated and collected. The classical texts of Ayurveda provide guidelines on the time of collection of raw drugs. Hence following these indications the oleoresin was collected in two seasons as per reference of Acharya Charaka and Susrutha in Hemantha rutu (Dec-Jan) and Vasantha rutu (April-May) respectively. Analytical studies revealed that the oleoresin collected in Vasantha rutu contained more tannin, resin, volatile matter, phenolic content, which are the active ingredients of the drug as compared to the oleoresin collected in Hemantha rutu .This is a preclinical work and further clinical study has to be done to prove efficacy of the seasonally collected samples.
PMCID: PMC3336301  PMID: 22557341
Objective: Medicinal plants organize an effective source of folk and modern medicine. Correct identification, authentication and quality control are essential to ensure safety, therapeutic potency, efficacy and reproducible quality of herbal medicines. The aim of this study is to use taxonomic method for authentication of traditional herbal drugs which are commonly sold in herbal shops in Iran. In this regard, twenty-seven cases of herbal drugs suspected to be adulterated were investigated.
Material and Methods: Crude raw material of herbal drugs was prepared from the various markets in Iran and was identified at the Ferdowsi University of Mashhad Herbarium (FUMH).
Results: Taxonomic evaluation revealed that 78 species belonging to 21 families which are traded in Iranian market should be considered as authentic, adulterated and substituted samples.
Conclusion: It was concluded that nowadays, many of the medicinal plants available in the market have ambiguous identification along with adulteration and contamination. The present study provides awareness amongst the traders, researchers, clinicians and manufacturing units about the ambiguity of authenticity in the traded herbal raw materials.
PMCID: PMC4075662  PMID: 25050238
Adulteration; Authentication; Crude Herbs; Identification; Iran; Market
BioMed Research International  2014;2014:735891.
The effectiveness of Okra gum in sustaining the release of propranolol hydrochloride in a tablet was studied. Okra gum was extracted from the pods of Hibiscus esculentus using acetone as a drying agent. Dried Okra gum was made into powder form and its physical and chemical characteristics such as solubility, pH, moisture content, viscosity, morphology study using SEM, infrared study using FTIR, crystallinity study using XRD, and thermal study using DSC and TGA were carried out. The powder was used in the preparation of tablet using granulation and compression methods. Propranolol hydrochloride was used as a model drug and the activity of Okra gum as a binder was compared by preparing tablets using a synthetic and a semisynthetic binder which are hydroxylmethylpropyl cellulose (HPMC) and sodium alginate, respectively. Evaluation of drug release kinetics that was attained from dissolution studies showed that Okra gum retarded the release up to 24 hours and exhibited the longest release as compared to HPMC and sodium alginate. The tensile and crushing strength of tablets was also evaluated by conducting hardness and friability tests. Okra gum was observed to produce tablets with the highest hardness value and lowest friability. Hence, Okra gum was testified as an effective adjuvant to produce favourable sustained release tablets with strong tensile and crushing strength.
PMCID: PMC3942280  PMID: 24678512

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