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   2015| October-December  | Volume 7 | Issue 4  
    Online since October 23, 2015

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Current status of herbal product: Regulatory overview
Sanjay Sharma
October-December 2015, 7(4):293-296
DOI:10.4103/0975-7406.168030  PMID:26681886
A review of the regulatory status of herbal drugs/products was done for few countries forming part of Asia, Africa, America, Europe, and Australia, to understand various categories under which the trade of herbal products is permitted and their premarketing requirements. A critical assessment was done, to know the hindrances in the process of harmonization of herbal products. It has been found that there is a lack of harmonization in the regulatory requirements of herbal products internationally, besides the issues of availability of herbs and their conservation. These are hindering the international trade and growth of the herbal products segment.
  4,474 274 -
Challenges and guidelines for clinical trial of herbal drugs
Abida Parveen, Bushra Parveen, Rabea Parveen, Sayeed Ahmad
October-December 2015, 7(4):329-333
DOI:10.4103/0975-7406.168035  PMID:26681895
World Health Organization (WHO) has defined herbal medicines as finished labeled medicinal product that contain an active ingredient, aerial, or underground parts of the plant or other plant material or combinations. According to a report of WHO, about 80% of the world population is reported to rely on traditional medicine for their primary health care needs. Even in the developed countries, complementary or alternative medicine is gaining popularity. A report of a global survey on national policy on traditional medicine and regulation of herbal medicines indicated that about 50 countries including China, Japan, and Germany already have their national policy and laws on regulations of traditional medicines. Herbal drugs possess a long history of its use and better patient tolerance. These are cheaper and easily available in countries like India due to rich agro culture conditions. However, reckless utilization of resources threatens the sustainability of several plant species. Traditional medicines are governed by the Drugs and Cosmetics Act of 1940 and the Drugs and Cosmetics Rules of 1945. In 1959, the Government of India amended the Drugs and Cosmetics Act to include drugs that are derived from traditional Indian medicine. In 1993, the guidelines for the safety and efficacy of herbal medicines developed by an expert committee directed that the procedures laid down by the office of the Drug Controller General of India for allopathic drugs should be followed for all traditional and herbal products to enter into clinical trials for any therapeutic condition. However, there are certain loop holes in the clinical trials of herbal drugs as the lack of stringent bylaws and regulations. Hence, a deep insight of important challenges and major regulatory guidelines for clinical trial of herbal drugs and botanicals is discussed in the present communication. There is lack of scientific evidence to evaluate safety and efficacy of herbal drugs. The quality of the trial drug has to be tested for batch-to-batch uniformity of the active constituents. It is very difficult to have active and control groups with identical color, smell and taste of the herbal drug, which cannot be imitated while manufacturing a placebo. These challenges can be reduced or overcome by applying most recent methodologies and guidelines for clinical trials. Since the quality control of herbal medicines is complicated and difficult, relevant and appropriate requirements should be established for the assessment of safety and efficacy for different categorized herbal medicines to reduce cost and expenditure. And, efforts should be made for the integration of traditional medicine into national healthcare systems. Different challenges and regulatory guidelines discussed for the clinical trial of herbal drugs will be useful for various industries for considering it before going ahead for clinical trial of their product.
  4,272 258 7
Optimization of ultrasound-assisted extraction of charantin from Momordica charantia fruits using response surface methodology
Javed Ahamad, Saima Amin, Showkat R Mir
October-December 2015, 7(4):304-307
DOI:10.4103/0975-7406.168032  PMID:26681889
Background: Momordica charantia Linn. (Cucurbitaceae) fruits are well known for their beneficial effects in diabetes that are often attributed to its bioactive component charantin. Objective: The aim of the present study is to develop and optimize an efficient protocol for the extraction of charantin from M. charantia fruits. Materials and Methods: Response surface methodology (RSM) was used for the optimization of ultrasound-assisted extraction (UAE) conditions. RSM was based on a three-level, three-variable Box-Behnken design (BBD), and the studied variables included solid to solvent ratio, extraction temperature, and extraction time. Results: The optimal conditions predicted by the BBD were: UAE with methanol: Water (80:20, v/v) at 46°C for 120 min with solid to solvent ratio of 1:26 w/v, under which the yield of charantin was 3.18 mg/g. Confirmation trials under slightly adjusted conditions yielded 3.12 ± 0.14 mg/g of charantin on dry weight basis of fruits. The result of UAE was also compared with Soxhlet extraction method and UAE was found 2.74-fold more efficient than the Soxhlet extraction for extracting charantin. Conclusions:A facile UAE protocol for a high extraction yield of charantin was developed and validated.
  2,798 73 -
Stability studies of silymarin nanoemulsion containing Tween 80 as a surfactant
Rabea Parveen, Sanjula Baboota, Javed Ali, Alka Ahuja, Sayeed Ahmad
October-December 2015, 7(4):321-324
DOI:10.4103/0975-7406.168037  PMID:26681893
Background: Silymarin, a flavonolignan from "milk thistle" (Silybum marianum) plant is used almost exclusively for hepatoprotection. Because of its low bioavailability, it was incorporated into a nanoemulsion formulation. The aim of the present study was to check the stability of silymarin nanoemulsion at different temperatures for 3 months. Materials and Methods: The oil-in-water based nanoemulsion formulation was prepared by titration method. Silymarin nanoemulsion was characterized by droplet size, viscosity, and refractive index. Droplet size, viscosity, and refractive index were determined every month. The shelf-life of silymarin nanoemulsion was determined by accelerated stability testing. Results: It was found that there was no significant change in the droplet size, viscosity, and refractive index at refrigerator and room temperature during the period of 3 months. The half-life of the optimized nanoemulsion formulation was found to be 4.74 years at room temperature. Conclusion: These results indicated that stability of silymarin can be enhanced in nanoemulsion formulation using Tween 80 as a surfactant.
  2,674 132 3
Quality control aspects of herbs and botanicals in developing countries: Coleus forskohlii Briq a case study
Ennus Tajuddin Tamboli, Karishma Chester, Sayeed Ahmad
October-December 2015, 7(4):254-259
DOI:10.4103/0975-7406.168020  PMID:26681877
Objective: Current trend of commercialization of herbal medicines draw a huge need of maintaining their quality. The declaration of quality, safety and efficacy of medicinal plants as well as poly-herbal formulations has become an important issue. Hence, qualitative and quantitative analysis of herbal drugs and formulations viz., fingerprint profiles and quantification of the various markers become key factors of quality control. Materials and Methods: Present investigation is a detailed report for quality control of well-known herb Coleus forskohlii Briq, which includes physicochemical parameter determination, safety evaluation, microscropical evaluation, and chromatographic fingerprinting as well. Results: Physico-chemical characters were evaluated according to Indian Pharmacopoeia, further microscopic evaluation of transverse section of Coleus reveals that periderm, secondary phloem, and wide secondary xylem cylinder, which occupies major portion of the root fragmentary. Chromatographic fingerprint profiles of Coleus have been generated, and a marker based standardization strategy was adopted; using different analytical technique like high-performance thin layer chromatography, high-performance liquid chromatography and gas chromatography-mass spectroscopy to maintain quality and ensure safety as well as efficacy. Conclusion: These advancements in modern techniques of analysis can lead to effective quality control of Coleus as well as other herbs. Present report can act as pioneer for quality control of modern herbal medicine.
  2,298 86 -
Development and validation of high-performance liquid chromatography and high-performance thin-layer chromatography methods for the quantification of khellin in Ammi visnaga seed
Abid Kamal, Washim Khan, Sayeed Ahmad, FJ Ahmad, Kishwar Saleem
October-December 2015, 7(4):308-313
DOI:10.4103/0975-7406.168033  PMID:26681890
Objective: The present study was used to design simple, accurate and sensitive reversed phase-high-performance liquid chromatography RP-HPLC and high-performance thin-layer chromatography (HPTLC) methods for the development of quantification of khellin present in the seeds of Ammi visnaga. Materials and Methods: RP-HPLC analysis was performed on a C18 column with methanol: Water (75: 25, v/v) as a mobile phase. The HPTLC method involved densitometric evaluation of khellin after resolving it on silica gel plate using ethyl acetate: Toluene: Formic acid (5.5:4.0:0.5, v/v/v) as a mobile phase. Results: The developed HPLC and HPTLC methods were validated for precision (interday, intraday and intersystem), robustness and accuracy, limit of detection and limit of quantification. The relationship between the concentration of standard solutions and the peak response was linear in both HPLC and HPTLC methods with the concentration range of 10–80 μg/mL in HPLC and 25–1,000 ng/spot in HPTLC for khellin. The % relative standard deviation values for method precision was found to be 0.63–1.97%, 0.62–2.05% in HPLC and HPTLC for khellin respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.53% in HPLC and 100.08% in HPTLC for khellin. Conclusions: The developed HPLC and HPTLC methods for the quantification of khellin were found simple, precise, specific, sensitive and accurate which can be used for routine analysis and quality control of A. visnaga and several formulations containing it as an ingredient.
  2,269 95 -
Development of standard operating procedure and standardization of Habb-e-Banafsha Qawi-A Unani polyherbal formulation
Athar Ali, Sabiha Sumbul, Malik Mobeen Ahmad, Sayeed Ahmad, Hifzul Kabir, MZ Abdin
October-December 2015, 7(4):250-253
DOI:10.4103/0975-7406.168019  PMID:26681876
Introduction: Herbals drugs became a boon for mankind since ancient times and still are used worldwide for the treatment of various human ailments. The safety of alternative medicinal preparations has been questioned due to reports of unwanted side effects. To maintain the quality and efficacy of these drugs, it is essential to standardize them in order to avoid the use of substandard or adulterated medicines in the market. Unani system of medicine mainly focuses on the treatment by natural drugs. Habb-e-Banafsha Qawi is useful in a cough, catarrah, and coryza. Materials and Methods: Physiochemical constants of Habb-e-Banafsha Qawi were determined through organoleptic characters, macro- and micro-scopic characters, ash value, solubility, pH values. Chromatographic fingerprints were developed using thin layer chromatography method. Aflatoxin (AF) contamination, heavy metal, and pesticide residues were also evaluated by standard methods.Objectives: In the present study, an attempt has been made to develop standard operating procedure and physiochemical parameters to monitor the quality of a Unani poly-herbal formulation, Habb-e-Banafsha Qawi. Results: The tablets tasted sweetish bitter with a pleasant odor, water soluble and acidic in nature. Rfvalues were almost same in all the extracts. No AF, heavy metal, and pesticide residues were observed. Conclusions: It may be concluded that the values and chromatographic fingerprints obtained can be used for quality evaluation and to set new pharmacopoeial standards for the preparation of Habb-e-Banafsha Qawi.
  2,222 112 -
Antidiabetic activity of ethanolic extract of Cyperus rotundus rhizomes in streptozotocin-induced diabetic mice
Pradeep Singh, Ratan L Khosa, Garima Mishra, Keshri K Jha
October-December 2015, 7(4):289-292
DOI:10.4103/0975-7406.168028  PMID:26681885
Objective: In the present investigation, ethanolic extract of Cyperus rotundus (EECR) rhizomes was evaluated for antidiabetic activity in streptozotocin (STZ)-induced diabetic swiss mice. Materials and Methods: After administration of EECR extract for 3 weeks, the body weight, blood glucose, biomarker enzymes (serum glutamic pyruvic transaminase [SGPT] and serum glutamic oxaloacetic transaminase [SGOT]), and plasma lipid levels were measured in STZ-induced diabetic mice. Results: The ethanolic extract at dose levels of 250 and 500 mg/kg body weight revealed significant antidiabetic activity, improvement in body weight, and reduction in elevated biochemical parameters such as SGPT, SGOT, cholesterol, and triglyceride levels. Conclusion: These experimental findings seemed to indicate the use of this plant in traditional Indian medicine for the treatment of diabetes.
  2,155 119 1
A high performance thin layer chromatographic method for the estimation of colchicine in different formulations
Mohd Fahim, Mhaveer Singh, YT Kamal, Hayat M Mukhtar, Sayeed Ahmad
October-December 2015, 7(4):260-263
DOI:10.4103/0975-7406.168021  PMID:26681878
Background: Colchicine is a main alkaloid present in bitter and sweet variety of colchicum (Colchicum luteum Baker), which have been reported to possess anti-rheumatic, anti-gout, and anticancer potential. Colchicum is an important ingredient of several Unani and Herbal formulations. Quantification of colchicine will play a great role in quality control of these formulations. Hence, a high-performance thin layer chromatographic (TLC) method has been developed for the analysis of colchicine in Unani formulations of various dosage forms such as hubb (tablet) and capsules. Materials and Methods: The samples were applied on aluminum TLC plates precoated with silica gel 60-F254and developed using mobile phase toluene-dichloromethane-methanol in equal proportions. Quantification was done by densitometric scanning at 350 nm, which showed a linear response in the range of 50–500 ng/spot. The developed method was validated as per the International Conference on Harmonization guidelines for linearity, precision, accuracy, specificity, robustness, limit of detection, and limit of quantification. Results and Conclusion: The developed method was applied for quantitative estimation of colchicine in different Unani and Herbal formulations. The method was found simple, selective, accurate with a wide range of linearity, hence suitable for the quality control of different formulations and varieties of colchicum with respect to colchicine content.
  2,154 106 -
Chromatographic analysis of wheatgrass extracts
Masood Shah Khan, Rabea Parveen, Kshipra Mishra, Rajkumar Tulsawani, Sayeed Ahmad
October-December 2015, 7(4):267-271
DOI:10.4103/0975-7406.168023  PMID:26681880
Aim: Wheatgrass (WG) is the shoot of Triticum aestivum Linn. belongs to the family Gramineae, and possess high chlorophyll content and essential vitamins, minerals, vital enzymes, amino acids, dietary fibers etc., It has been shown to possess anti-cancer, anti-ulcer, antioxidant, and anti-arthritic activity due to the presence of biologically active compounds, and minerals. Therefore, in the present study, high-performance thin layer chromatography (HPTLC), and high-performance liquid chromatography (HPLC) methods for qualitative and quantitative analysis have been proposed, which will help in quality evaluation of wheat grass extract. Materials and Methods: Samples for analysis were prepared in methanol and water simply by sonication. These were applied on pre-coated silica plate and chromatograms were developed using toluene: Ethyl acetate: Formic acid. HPLC analysis was done on Waters HPLC system using water, methanol, and acetonitrile as mobile phase. Merck C18 column has been used. Results: HPTLC finger printing of alcoholic extracts of WG was carried out and found 10–11 spots at different wavelengths 254, 366, and 435 nm. HPLC fingerprinting produced 22 peaks at 256 nm. Quantitative HPTLC analysis was done to determine the gallic acid content, and was found to be 0.077% w/w in aqueous extract. By HPLC, the content of gallic acid and rutin was found to be 0.07%, and 0.04% w/w in aqueous extract of WG. Conclusion: The developed HPLC and HPTLC fingerprinting method can be used for the quality control, and standardization of WG and its extracts used as nutritional supplement.
  2,129 102 -
Determination of nucleosides in Cordyceps sinensis and Ganoderma lucidum by high performance liquid chromatography method
Masood Shah Khan, Rabea Parveen, Kshipra Mishra, Rajkumar Tulsawani, Sayeed Ahmad
October-December 2015, 7(4):264-266
DOI:10.4103/0975-7406.168022  PMID:26681879
Background: Nucleosides are supportive in the regulation and modulation of various physiological processes in body, they acts as precursors in nucleic acid synthesis, enhance immune response, help in absorption of iron and influence the metabolism of fatty acids. Cordyceps sinensis and Ganoderma lucidum are well-known for its use in traditional medicine of China, Nepal and India. They are rich in nucleosides such as adenine, adenosine, cordycepin, etc. Hence, a simple, economic and accurate high-performance liquid chromatography (HPLC) analytical method was proposed for determination of adenine and adenosine for the quality control of plants. Materials and Methods: Chromatographic experiments were conducted on YL9100 HPLC system (South Korea). Reversed-phase chromatography was performed on a C18 column with methanol and dihydrogen phosphate as the mobile phase in isocratic elution method at a flow rate of 1.0 mL/min. Detection was carried out at 254 nm, which gives a sharp peak of adenine and adenosine at a retention time of 6.53 ± 0.02 min and 12.41 ± 0.02, respectively. Results and Discussion: Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 25–200 µg/mL for adenosine and 100–800 µg/mL for adenine with regression coefficient of 0.999 and 0.996, respectively. The adenine was found 0.16% and 0.71% w/w in G. lucidum and in C. sinensis, respectively, and adenosine was found to be 0.14% w/w in G. lucidum whereas absent in C. sinensis. Conclusion: The developed HPLC method for the quantification of adenosine and adenine can be used for the quality control and standardization of crude drug and for the different herbal formulations, in which adenine and adenosine are present as major constituents. The wide linearity range, sensitivity, accuracy, and simple mobile phase imply the method is suitable for routine quantification of adenosine and adenine with high precision and accuracy.
  2,131 73 -
Hepatoprotective activity of a new polyherbal formulation against paracetamol and D-galactosamine induced hepatic toxicity
Mohammad Ahmed Khan, Arun Gupta, Satyendra Kumar, Sayeed Ahmad, J.L.N. Sastry
October-December 2015, 7(4):246-249
DOI:10.4103/0975-7406.168018  PMID:26681875
Objective: The present study was envisaged to evaluate the protective effect of polyherbal formulation, DRDC/AY/8060, developed by Dabur India Ltd., against paracetamol and D-galactosamine induced hepatic toxicities in Wistar rats. Materials and Methods: The study was carried out in two different experiments of 10 and 14 days against paracetamol and D-galactosamine, respectively. Animals were divided into different treatment groups (n = 6). The control group received normal saline, a toxicant group in two experiments received paracetamol 750 mg/kg p.o. every 72 h for 10 days and D-galactosamine 400 mg/kg i.p. single dose. The test formulation was used at the two dose levels of 120 and 240 mg/kg/day. Treatment groups treated with test formulations were also administered D-galactosamine as given in toxicant group. At the end of the dosing schedule, blood was withdrawn from the retrobulbar plexus of the animals for serum estimation of serum glutamate oxaloacetate transferase (SGOT), serum glutamate pyruvate trasnferase (SGPT), albumin, bilirubin, and alkaline phosphatase (ALP). Following the withdrawal of blood animals was sacrificed, and liver tissue was excised for estimation of thiobarbituric acid reactive substances (lipid peroxidation, malondialdehyde), tissue glutathione (GSH) and histopathological studies. Results: It was evident from the biochemical estimation that both paracetamol and galactosamine caused hepatotoxicity in the toxicant groups. However, treatment with DRDC/AY/8060 significantly (P < 0.001, vs. toxicant) reduced the levels of SGOT, SGPT, serum bilirubin, and ALP, as well as decreased lipid peroxidation. In addition, treatment with test formulation also significantly (P < 0.001, vs. toxicant) elevated serum albumin and GSH levels compared to toxicant groups. Conclusion: On the basis of these studies and comparative evaluation it can be concluded that the formulation DRDC/AY/8060 showed hepatoprotective activity against paracetamol and D-galactosamine at 120 mg/kg and 240 mg/kg.
  2,032 129 1
Evaluation of anticancer potential of Bacopa monnieri L. against MCF-7 and MDA-MB 231 cell line
Md. Nasar Mallick, Md. Salman Akhtar, Mohd Zeeshan Najm, ET Tamboli, Sayeed Ahmad, Syed Akhtar Husain
October-December 2015, 7(4):325-328
DOI:10.4103/0975-7406.168038  PMID:26681894
Background: The ethanolic extract of Bacopa monnieri contains bacoside A and B, brahmin, cucurbitacins, and betulinic acid. Currently, cucurbitacins have also been reported for their strong anti-tumorigenic and anti-proliferative activity by inducing cell cycle arrest at the G2/M phase and formation of multiplied cells. The present study was carried out to evaluate the in vitro cytotoxic activity of ethanolic extract of dichloromethane (DCM) fraction of B. monnieri on two different cell lines. Materials and Methods: The ethanolic extract of B. monnieri was prepared using soxhlet extraction method and different fractions (hexane, DCM, methanol, acetone, and water) of ethanolic extracts were prepared. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay of ethanolic extract and of all fractions was carried out on MCF-7 and MDA-MB 231 cell lines. The presence of cucurbitacins and betulinic acid in these fractions was confirmed by high-performance thin layer chromatography. Results: The IC50 values of ethanolic extract of B. monnieri in MCF-7 and MDA-MB 231 cell lines were 72.0 μg/mL and 75.0 μg/mL, respectively. The DCM fraction of B. monnieri showed maximum cytotoxic activity among all fraction upto 72 h and was found to be 57.0 μg/mL and 42.0 μg/mL, respectively. Conclusion: The results showed good cytotoxic activity in DCM fraction in both the cell lines may be due to the presence of cucurbitacins and betulinic acid in DCM fraction.
  2,019 134 -
Standardization and in vitro antioxidant activity of jatamansi rhizome
Mhaveer Singh, Mohammad A Khan, Masood S Khan, SH Ansari, Sayeed Ahmad
October-December 2015, 7(4):275-279
DOI:10.4103/0975-7406.168025  PMID:26681882
Background: 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. Results: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. Conclusion: The findings of the present research can be utilized for the identification and quality control of the jatamansi rhizome.
  2,022 100 -
Effect of piperine on pharmacokinetics of sodium valproate in plasma samples of rats using gas chromatography-mass spectrometry method
Bushra Parveen, KK Pillai, ET Tamboli, Sayeed Ahmad
October-December 2015, 7(4):317-320
DOI:10.4103/0975-7406.168036  PMID:26681892
Piperine (PIP) is used as anticonvulsant in traditional Chinese medicine. Co-administration of low-dose sodium valproate with PIP has been regarded to have potential anticonvulsant activity. Aim: This study was intended to investigate the effect of PIP on the pharmacokinetics of sodium valproate (SVP) in the plasma samples of rats using gas chromatography-mass spectrometry (GC-MS) method. Materials and Methods: The plasma samples obtained after oral administration of SVP, 150 mg/kg and SVP, 150 mg/kg + PIP, and 5 mg/kg to male Wistar rats were used to quantify the concentrations in plasma using GC-MS method. Results: A simple and accurate method developed in-house was applied for the analysis of plasma samples of Wistar rats after oral administration of SVP and PIP + sodium valproate, respectively. The pharmacokinetic parameters reported 14.8-fold increase in plasma concentration (maximum observed concentration in the concentration-time profile), 4.6-fold increase in area under the curve and slightly prolonged time to reach that concentration (1 h) of SVP in presence of PIP. Conclusion: The study reaffirms the bioenhancing effect of PIP suggesting possibility of dose reduction of SVP while co-adminstering with PIP.
  1,875 106 3
Antimicrobial activity of aqueous and organic extracts of a Saudi medicinal plant: Rumex nervosus
Abdul Rahman K Al-Asmari, Yunus M Siddiqui, Md Tanwir Athar, Ahmed Al-Buraidi, AS Al-Eid, Ghalib B Horaib
October-December 2015, 7(4):300-303
DOI:10.4103/0975-7406.168031  PMID:26681888
Objective: The antimicrobial effect of aerial part of Rumex nervosus obtained from the Southern region of Saudi Arabia was evaluated on bacterial strains Staphylococcus aureus, methicillin resistant S. aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas Aeruginosa, and fungal strain Candida albicans. Materials and Methods: The solvents used for the extraction were aqueous, hexane, and methanol. The in vitro antimicrobial activity was performed by agar diffusion and disk diffusion methods and the confirmation of this activity was done by the enumeration of colony forming units (CFU). Results: The aqueous extract showed the growth inhibitory effect on Gram-positive bacteria while the Gram-negative P. aeruginosa was the most sensitive microorganism as determined by the agar diffusion technique. Surprisingly, the extract showed little antibacterial activity on other Gram-negative bacteria (E. coli) by this technique. Ethanolic extract was also found to be inhibitory to the growth of microorganisms. Hexane extract was relatively low in antimicrobial activity on Gram-negative E. coli and P. aeruginosa, while both the organic extracts were inhibitory to the growth of the fungus, C. albicans. Hexane gave no conclusive results with agar or disk diffusion methods, but showed the microbial growth inhibition in CFU enumeration. The antibacterial activity of active extracts was compared with vancomycin while antifungal activity of was compared with amphotericin B. Conclusion: The results obtained in the present study suggest that R. nervosus showed a marked antimicrobial activity with the test organisms.
  1,902 57 -
Quality control and in vitro antioxidant potential of Coriandrum sativum Linn.
Mhaveer Singh, ET Tamboli, YT Kamal, Wasim Ahmad, SH Ansari, Sayeed Ahmad
October-December 2015, 7(4):280-283
DOI:10.4103/0975-7406.168026  PMID:26681883
Background: Coriandrum sativum Linn., commonly known as coriander, is a well-known spice and drug in India. It has various health-related benefits and used in various Unani formulations. In this present study, quality assessment of coriander fruits was carried out by studying anatomical characters, physicochemical tests, and chemoprofiling using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectroscopy (GC-MS) along with in vitro antioxidant potential. Materials and Methods: Standardization was carried out as per the pharmacopeial guidelines. Estimation of heavy metals, pesticides, and aflatoxins was carried out to ascertain the presence of any contaminant in the sample. Chemoprofiling was achieved by thin layer chromatography (TLC) by optimizing the mobile phase for different extracts. The most of the pharmacological activities of coriander are based on volatile oil constituents. Hence, GC-MS profiling was also carried out using hexane-soluble fraction of hydro-alcoholic extract. The total phenolic contents and in vitro antioxidant efficacy were determined using previously established methods. Results: The quality control and anatomical studies were very valuable for the identification whereas good antioxidant potential was observed when compared to ascorbic acid. The drug was found free of contaminant when analyzed for pesticides and aflatoxins whereas heavy metals were found under reported limits. Conclusion: The work embodied in this present research can be utilized for the identification and the quality control of the coriander fruit.
  1,843 86 1
Characterization of Linum usitatissimum L. oil obtained from different extraction technique and in vitro antioxidant potential of supercritical fluid extract
Rishika Chauhan, Karishma Chester, Yasmeen Khan, Ennus Tajuddin Tamboli, Sayeed Ahmad
October-December 2015, 7(4):284-288
DOI:10.4103/0975-7406.168027  PMID:26681884
Aim: Present investigation was aimed to characterize the fixed oil of Linum usitatissimum L. using five different extraction methods: Supercritical fluid extraction (SFE), ultrasound-assistance, soxhlet extraction, solvent extraction, and three phase partitioning method. Materials and Methods: The SFE conditions (temperature, pressure, and volume of CO2) were optimized prior for better yield. The extracted oils were analyzed and compared for their physiochemical parameters, high performance thin layer chromatography (HPTLC), gas chromatography-mass spectrometry (GC-MS), and Fourier-transformed infrared spectroscopy (FT-IR) fingerprinting. Antioxidant activity was also determined using 1,1-diphenyl-2-picrylhydrazyl and superoxide scavenging method. Result: The main fatty acids were α-linolenic acid, linoleic acid, palmitic acid, and stearic acid as obtained by GC-MS. HPTLC analysis revealed the presence of similar major components in chromatograms. Similarly, the pattern of peaks, as obtained in FT-IR and GC-MS spectra of same oils by different extraction methods, were superimposable. Conclusion: Analysis reported that the fixed oil of L. usitatissimum L. is a good source of n-3 fatty acid with the significant antioxidant activity of oil obtained from SFE extraction method.
  1,772 75 -
Effect of detoxification (tadbeer) in content of toxic metabolites of Strychnos nux-vomica: A Unani approach for its use in human
Noman Anwar, Masood Shah Khan, Hifzul Kabir, Sayeed Ahmad
October-December 2015, 7(4):314-316
DOI:10.4103/0975-7406.168034  PMID:26681891
Background: Azaraqi (Strychnos nux-vomica, Loganiaceae) has been the important Unani medicine since long time as a stimulant, anti-inflammatory, and blood purifier. It has been used very frequently by the Unani practitioner. But the Unani system recommends application of azaraqi in medicine only after its detoxification (tadbeer) may be because of the presence of its deadly poisonous alkaloids (strychnine and brucine). In the present investigation, an attempt has been made to quantify the actual content of their toxic alkaloids before and after the tadbeer. Materials and Methods: A sensitive high-performance thin layer chromatographic (HPTLC) method has been developed for estimation of strychnine and brucine in different samples of azaraqi before and after tadbeer. Precoated HPTLC silica gel plates were used as stationary phase and (toluene: Ethyl acetate: Dietylamine 7:2:1 v/v/v) was used as mobile phase. Result: The Rfvalue of strychnine and brucine was found as 0.53 and 0.41, respectively. Detection and quantification were performed by densitometry at 270 nm. The calibration plot was linear in the range of 50–1000 ng of strychnine and brucine, respectively, with the correlation coefficient (r2) 0.993 and 0.991 for strychnine and brucine, respectively, which confirms good linearity. The content of strychnine was 0.175, 0.07, 0.18, 0.051, and 0.075% w/w whereas brucine was 0.16, 0.117, 0.061, 0.045, and 0.057 in crude azaraqi, azaraqi without outer cover, azaraqi outer cover only, azaraqi mudabbar and azaraqi mudabbar by fried in ghee, respectively. Conclusion: The detoxification results in sharp decrease in content of toxic metabolites. The process by boiling in milk was found much effective but tedious as compare to frying method.
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Hepatoprotective potential of kumaryasava and its concentrate against CCl4-induced hepatic toxicity in Wistar rats
Mohammad Ahmed Khan, Arun Gupta, J. L. N Sastry, Sayeed Ahmad
October-December 2015, 7(4):297-299
DOI:10.4103/0975-7406.168029  PMID:26681887
Objective: Kumaryasava (KS) is a marketed Ayurvedic formulation containing Aloe vera as the main ingredient. It has been used widely for the treatment of liver disorders; however, there is a lack of modern scientific data on hepatoprotection. The recommended dose of KS is high and up to 60 mL/day. The present study describes the preparation of new KS concentrate and evaluation of comparative hepatoprotective activity of KS and prepared KS concentrate at one-third of KS dose against CCl4-induced hepatic toxicity. Materials and Methods: Animals were divided into different groups (n = 6). The first group received normal saline (control) 1.0 mL/Kg/day p.o. for 10 days. The second group (toxicant) was given normal saline 1.0 mL/Kg/day p.o. for 10 days with CCl4 in olive oil (1:1 v/v) at 1.0 mL/Kg/day p.o. Third, fourth, and fifth groups received KS, KS concentrate and a marketed formulation as standard) at doses of 5.0 mL/Kg/day p.o., 1.6 mL/Kg/day p.o., and 100 mL/Kg/day p.o. (tablet suspended in water using 0.1% carboxymethyl cellulose) respectively for 10 days along with CCl4 as given to the toxicant group. On the 11th day, blood was withdrawn from retro-orbital plexus and serum was separated for biochemical estimation of serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP), and albumin levels. Later, animals were sacrificed under high dose of anesthesia to remove liver tissue, which were removed and washed with ice cold saline for the estimation of lipid peroxidation. Liver tissue from each group was also fixed in 10% formalin for histopathological analysis. Results: Results demonstrated that both KS and KS concentrate showed the protection against CCl4-induced hepatic toxicity. This was evident from the reduction in serum SGOT, SGPT, ALP levels, and elevation in serum albumin levels observed post treatment of CCl4 treated rats with KS and KS concentrate, which were supported by histopathological data. Conclusion: KS concentrate can be a useful hepatoprotective formulation which may help in reducing the high dose of KS to approximately one-third of the recommended dose.
  1,639 82 1
National seminar on "regulatory and quality aspects of herbal drugs and botanicals"
Sayeed Ahmad
October-December 2015, 7(4):245-245
DOI:10.4103/0975-7406.168039  PMID:26681874
  1,602 100 -
Bergenin determination in different extracts by high-performance thin-layer chromatographic densitometry
Masood Shah Khan, Washim Khan, Wasim Ahmad, Mhaveer Singh, Sayeed Ahmad
October-December 2015, 7(4):272-274
DOI:10.4103/0975-7406.168024  PMID:26681881
Aim: Bergenin is an active secondary metabolite, found in Bergenia ligulata, family Saxifragaceae, which is an important medicinal plant used in the traditional system of medicine. It is distributed throughout the South and East Asia and some European countries, usually growing on high altitude in the Himalayan region and known as Pashanbhed (meaning "to break the stone"). The rhizome of B. ligulata has been used since long time in different traditional formulations of kidney and liver disorders. Due to its exhaustive use in the traditional system, it is commonly adulterated with the rhizome of other plants which do not contain its chemical marker bergenin. Hence, we developed high-performance thin-layer chromatographic (HPTLC) method for quantification of bergenin in B. ligulata which can be used for its quality control. Materials and Methods: A sensitive HPTLC method has been developed for the estimation of bergenin in different extracts of B. ligulata and its traditional formulations. Precoated HPTLC silica gel plates were used as stationary phase, and chloroform: methanol: acetic acid (8:1:1, v/v/v) was used as mobile phase. Results: The Rfvalue of bergenin was found to be 0.28 ± 0.03. Detection and quantification were performed by densitometry at 276 nm. The calibration plot was linear in the range of 200–5000 ng of bergenin with the correlation coefficient of (r2) 0.999, which confirms good linearity. The content of bergenin in methanolic and acetone extracts was found to be 5.51 ± 0.14 and 5.76 ± 0.16, respectively. Conclusion: The method can be applied for quality control and standardization of B. ligulata and its traditional formulations as well as for checking the presence of adulterants.
  1,585 77 -