Pharmacognosy Research

Secondary metabolites from lichen Usnea longissima and its pharmacological relevance Moumita Dandapat, Santanu Paul Pharmacognosy Research 2019 11(2):103-109 The objective of the study was to isolate chemical compounds from lichen Usnea longissima (UL) and to use lichen metabolites as antibiotics, antifungal, anti-HIV, anticancer, antiprotozoan, and antibacterial agents. Preparation of the lichen extracts: Fine powder of UL lichen was extracted using acetone, methanol, and water in a Soxhlet extractor. In a rotary evaporator, the extracts were filtered and then concentrated under reduced pressure. The dry extracts were stored at 18°C until they were used in the tests. The extracts were used for various assays. Sixteen compounds were isolated from lichen UL using various chromatographic techniques, including silica gel, Sephadex LH-20, operational data store, and semipreparative high-performance liquid chromatography. By spectroscopic data analyses, their structures were identified as (1) useanol, (2) lecanorin, (3) 3-hydroxy-5-methylphenyl 2-hydroxy-4-methoxy-6-methylbenzoate, (4) lecanorin E, (5) 3'-methylevernic acid, (6) evernic acid, (7) barbatinic acid, (8) 3,7-dihydroxy-1,9-dimethyldibenzofuran, (9) orcinol, (10) O-methyl orcinol, (11) methyl orsellinate, (12) methyl everninate, (13) 2,5-dimethyl-1,3-benzenediol, (14) 2-hydroxy-4-methoxy-3,6-dimethyl benzoic acid, (15) ethyl everninate, and (16) ethyl 2,4-dihydroxy-6-methylbenzoate. UL lichen is a great source of secondary metabolites, which can be used as antimicrobial, antiproliferative, and anticancer agents. This lichen can be used as a natural antioxidant or antitumor agent in clinical practice; commercially, it can be used as an anticancer drug.

Biological screening of tri-jannarose as a recipe from Thai traditional medicine Ladachart Taepongsorat, Ampa Konsue Pharmacognosy Research 2019 11(2):110-114 Context: Tri-Jannarose (TJ) is a Thai traditional medicine recipe, the ingredients of which are betel palm seed (Areca catechu L.), Siamese neem tree root (Azadirachta indica A. Juss.), and heart-leaved moonseed vines (Tinospora cordifolia [Thunb.] Miers). The equal mixture of three plants indicated to treatment of antipyretic, diuretic, expectorate, nourishment and appetizing. Aims: Phytochemical screening, antioxidant, and α-glucosidase inhibitory activities of TJ using different extractions were evaluated. Materials and Methods: The three plants of recipe were extracted using different solvents such as aqueous extract (ATJ), 50% ethanolic extract (HETJ), and 95% ethanolic extract (ETJ). The phytochemical screening was determined by total phenolic compounds and total flavonoid contents. The anti-oxidation were tested using by 2,2-diphenyl-1-picrylhydrazy (DPPH) radical scavenging and 2,2 -azinobis-(3-ethylbenzothiazoline-6-sulphonate) (ABTS+) assay. The α-glucosidase inhibitory activity was determined for glucose transferase mechanism. Results: Phytochemical screening found that this recipe had both phenolic and flavonoid substances. The ETJ (IC50 = 0.0463 ± 0.002) was exerted on antioxidation higher than HETJ and ATJ (IC50 = 0.0511 ± 0.000 and 0.1485 ± 0.005 mg/mL). Surprisingly, ABTS+ assay, ETJ (IC50 = 0.015 ± 0.000 mg/mL), and HETJ (IC50 = 0.022 ± 0.000 mg/mL) showed high effect on free radical scavenging activity than the standard controls, ascorbic acid (IC50 = 0.025 ± 0.001 mg/mL), and Trolox (IC50 = 0.032 ± 0.001 mg/mL). The α-glucosidase inhibitory activity found that all of the extract including ATJ (IC50= 0.0127±0.02 mg/mL), ETJ (IC50= 0.0154±0.01 mg/mL) and ETJ (IC50= 0.0202±0.01 mg/mL) were more potent to inhibit α-glucosidase emzyme than acarbose (IC50=0.745±0.026 mg/mL) as a positive control. Conclusion: The pharmaceutical preliminary scarring was confirmed to treatment on Thai traditional medicine. The recipe composed with phenolic compounds and flavonoids contents which chemical substance were more potent anti-oxidation, and bittersweet flavor was stronger to α-glucosidase inhibitory activity.

Antidiabetic activity of hydroalcoholic extract of Myrtus communis (myrtle) fruits in streptozotocin-induced and dexamethasone-induced diabetic rats Mohammad Sharif Talebianpoor, Mohammad Shafee Talebianpoor, Mahboubeh Mansourian, Tahereh Vafaiee-Nejad Pharmacognosy Research 2019 11(2):115-120 Background: Diabetes mellitus is characterized by an increase in blood glucose levels, resulting in insufficiency of insulin secretion, insulin resistance, or both. Objective: The effects of hydroalcoholic extract of the Myrtus communis (myrtle) fruits on Wistar rats were investigated in two types of diabetes mellitus. Materials and Methods: Intraperitoneal administration of streptozotocin (60 mg/kg) was used to induce Type I diabetes. Type II diabetes was induced by subcutaneous injection of 1 mg/kg/day dexamethasone for 10 days. Two groups of the diabetic animals received the hydroalcoholic extract of the fruit by gavage (250 mg/kg and 500 mg/kg for 45 and 10 days) and the diabetic control groups receiving distilled water. Results: The hydroalcoholic extract of M. communis fruits reduced the serum levels of the glucose, triglyceride, urine volume, urine protein, and malondialdehyde at the end of the 45 day. In Type II diabetic rats, there was a significant effect on plasma glucose levels. On this day, blood glucose-lowering effect was significantly observed after insulin administration as C2>D2M500>D2M250>D2 (P ≤ 0.05). The plasma level of insulin was completely reversed. There were no differences in the other biochemical parameters. Conclusions: The hydroalcoholic extract of M. communis fruits has a significant effect on the improvement of diabetes mellitus complications especially Type II diabetic animals, which begins with insulin resistance.

Identification of bioactive diterpenoid lactones using nuclear magnetic resonance from Andrographis paniculata Raghava N Sriramaneni, Amirin Sadikun, Naveen Kumar Hawala Shivashekaregowda, Annegowda Venkatappa, Mohamad Zaini Asmawi Pharmacognosy Research 2019 11(2):121-126 Background: Andrographis paniculata (AP) (Burm. F) Nees, belongs to the family Acanthaceae, it is also known as a highly bitter plant. In South East Asia this plant is commonly used for cold, cough and respiratory tract infections. Andrographolide (ANG), 14-deoxy-11, 12-didehydroandrographolide (DDA) are the major bioactive diterpenoid lactone compounds of the plant shown vasorelaxant effect, anticancer, anti-inflammatory, and to treat variety of diseases which is well-documented. Materials and Methods: The high-performance liquid chromatography (HPLC) and 1H-NMR analysis of AP chloroform extract (APCE) revealed the presence of ANG, 14-deoxyandrographolide (DA), and DDA. The main advantage of 1H-NMR is simple, rapid and successfully applied to quantify the active diterpenoids and an alternative to HPLC method to check the presence of various bioactive compounds and its presence in the active raw materials of AP. Results: The results revealed that most active bioactive compounds such as ANG, DA, and DDA are present in the APCE portion of the AP. The chemical shifts of various groups confirmed through 1H-NMR that all the three ANG, DA, and DDA has potent vasorelaxant effect and majority of these compounds are present in the APCE portion and not in the methanol or aqueous fractions. The 1H-NMR study confirmed that diterpenoids such as ANG, DA, and DDA from APCE has vasorelaxant effect on the animal study.

In vitro screening for antioxidant and antimicrobial properties of three lebanese medicinal plants crude extracts Josiane Alhage, Hoda Elbitar Pharmacognosy Research 2019 11(2):127-133 Background: Despite its small geographic area, Lebanon is characterized by a natural wealth in medicinal plants. Lebanese flora has about 2600 species, where >100 are endemic and are used in traditional medicine for their therapeutic effects. Objective: To evaluate the antioxidant and antimicrobial activities of dichloromethane, methanol, and aqueous crude extracts of three Lebanese medicinal plants: Geranium robertianum, Asphodelus microcarpus, and Alcea setosa. Materials and Methods: The antioxidant activity of different crude extracts was determined using the free radical, 1,1-diphenyl-2-picrylhydrazyl. Antimicrobial activity was evaluated in a plate-hole diffusion assay against two Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacterial strains, one Gram-positive (Staphylococcus aureus) bacterial strain, and a fungal strain (Candida albicans). Results: Methanol and aqueous extracts of G. robertianum possessed high scavenging activity (Radical scavenging activity (RSA) >85%) at 0.3 mg/mL followed by the extracts of A. microcarpus which showed a moderate activity. G. robertianum extracts exhibited good inhibition diameters against S. aureus growth. Conclusion: Obtained results give an overall view on the bioactivities of three Lebanese medicinal plants crude extracts.

Phytochemical and antimicrobial evaluation of Lauridia tetragona (L.F) R.H. Archer: A medicinal plant used for the management of dysentery in the Eastern Cape Province of South Africa Olubunmi Abosede Wintola, Anthony Jide Afolayan Pharmacognosy Research 2019 11(2):134-139 Objective: The present investigation evaluated the in vitro phytochemical and antimicrobial potential of the methanol and acetone leaf extracts of Lauridia tetragona (L.F), a plant consumed as a herb in South Africa. Materials and Methods: Spectrophotometry assays using Folin–Ciocalteu, aluminum chloride, and vanillin–hydrochloric acid were used for the determination of phenols, flavonoids, and proanthocyanidin contents of the extracts, respectively. The agar dilution technique was used for the antimicrobial evaluation. The inhibitory activity of the extracts was tested on five Gram-negative and five Gram-positive bacteria and four fungi. Results: The methanol extract had a higher phenol (54.87 ± 4.01 mg gallic acid equivalent [GAE]/g) and proanthocyanidin (78.55 ± 0.83 mg catechin equivalent [CE]/g) content than the acetone extracts with 45.27 ± 3.93 mg GAE/g and 63.54 ± 1.67 mg CE/g, respectively. The acetone extract, however, had higher flavonoid content (462.45 ± 1.93 mg quercetin equivalent [QE]/g) than the methanol extract (412.20 ± 3.85 mg QE/g). The minimum inhibitory concentration (MIC) of the antibacterial assay ranged from 2.5 to 5 mg/mL. All the bacteria except Staphylococcus aureus were susceptible to the acetone extract. Five bacteria, three Gram positive and two Gram negative, were resistant while the remaining five were susceptible to the methanol extract. Conversely, all the fungi tested were susceptible to both extracts with a MIC which ranged from 0.63 to 10 mg/mL. Conclusion: The results obtained revealed that Gram-positive and Gram-negative bacteria and fungi showed some degree of susceptibility to the plant extracts. This gives an indication of broad-spectrum activity exhibited by the crude extracts of L. tetragona and supports its ethnomedicinal usage.

Biochemical screening and determination of bioactive components of commercially cultured pacific white shrimp Penaeus vannamei Jayalakshmi Muniyappan, Vanitha Varadharajan, Pushpabharathi Namadevan Pharmacognosy Research 2019 11(2):140-146 Background: Marine waste is an extraordinarily renewable aid for the restoration of several valued metabolites with potential biological applications. Objectives: The investigation is planned to detect the biochemical components present in the prawn shell waste by qualitative, quantitative methods, to assess the antioxidant potential, and also to find the bioactive compounds existing in the prawn shell waste by gas chromatography-mass spectrometry (GC-MS) analysis. Materials and Methods: Penaeus vannamei shell wastes are collected, cleaned, dried, and powdered well. The bioactive compounds present in crude ethyl acetate extract of P. vannamei shell was determined by qualitative, quantitative, and GC-MS analysis. The free radical scavenging activity of the extract was studied by different in vitro antioxidant assays. Results: The bio-compounds such as carbohydrates, saponins, flavonoids, tannins, and quinones show the positive result by qualitative analysis. The higher tannin content 49.2 ± 0.084 mg/g was observed in the ethyl acetate extract of P. vannamei shell and the flavonoid was found to be 5.994 ± 0.044 mg/g. The GC-MS analysis of the P. vannamei shell shows the various numbers of bio-compounds. Some of the identified compounds are Timonacic which has the powerful antioxidant property, Octadecane, 3ethyl5(2ethylbutyl) is a good antifungal agent, Acetamide possesses antioxidant and anti-inflammatory property. The results of the in vitro assays revealed that P. vannamei shell extract possess significant antioxidant activity. Conclusion: The present study suggests that the effective utilization of prawn shell waste enhances biomedical research field for the development of the natural drug for many chronic diseases with no side effects and at the same time can reduce environmental pollution.

Characterization of the gain and loss of resistance to antibiotics versus tolerance to honey as an antimutagenic and antimicrobial medium in extended-time serial transfer experiments Salma G Abdel-Azim, Ahmad G Abdel-Azim, Brian P Piasecki, Gamal A Abdel-Azim Pharmacognosy Research 2019 11(2):147-154 Background: Honey contains several substances with antimicrobial properties that appear more recalcitrant to generating bacterial-evolved resistance than traditional antibiotics. Objectives: This study seeks to characterize the evolution of bacteria grown for successive generations in honey as an antimutagenic medium versus ampicillin. Materials and Methods: A naive strain of Staphylococcus aureus was serially cultured for 17 days in Lysogeny broth (LB) containing sublethal concentrations of medical-grade Manuka honey, polyfloral honey, and ampicillin. Glucose as an osmotic control and pure LB were included for comparison. A portion of each culture was removed every 24 h to (a) determine the amount of growth that occurred during the previous 24 h and (b) use as a genetic stock in serially transferred tubes containing the same inhibitory compound as used previously. Results: As indicated by an increase in growth over sequential 24-h period, bacteria rapidly gained resistance to ampicillin in a step-wise pattern. However, bacteria grown in Manuka and polyfloral honey never exceeded their initial growth levels. Bacteria grown in relatively high concentrations of honey for a single 24-h period consistently lost viability after one transfer, a phenomenon that has not been reported in the literature before and which indicates the inability of bacteria to adapt to honey as an antimutagenic medium. While bacteria grown in honey did not evolve the ability to grow at higher concentrations, a single isolate grown in Manuka honey gained tolerance to Manuka by successfully surviving and proliferating beyond second transfers. Bacteria that developed antibiotic resistance were found to remain sensitive to honey. Moreover, bacteria lost their resistance to ampicillin upon a single exposure to Manuka honey.

Evaluation of acute toxicity of plants' mixture used in traditional treatment of kidney diseases in Morocco Mohamed Chebaibi, Dalila Bousta, Laila Chbani, Imane Iken, Sanae Achour Pharmacognosy Research 2019 11(2):155-161 Background: The use of plants' mixture in the traditional treatment of kidney diseases in Morocco is widespread. Objective: To evaluate the toxic effects of plants' mixture used in the traditional treatment of kidney diseases in Morocco. Materials and Methods: The phytochemical screening was performed. For acute toxicity, single doses of low dose (300 mg/kg), medium dose (500 mg/kg), high dose (2000 mg/kg), lethal dose (6000 mg/kg), and traditional dose (10 ml/kg) body weight of aqueous extracts of plants' mixture were administered orally in Wistar rats. Animals were monitored daily for at least 15 days after an oral administration of aqueous extract of the mixture to detect any changes in body weight, behavior, autonomic profiles, or mortality. Calculation of relative organ weight (ROW), hematological, biochemical analysis, and histopathology evaluation were carried out. Results: The acute oral toxicity study showed diarrhea, somnolence, and agitation of different groups of rats, while no mortality and no statistically significant decrease in body weight was observed. Statistically, the kidneys, liver, and spleen showed significant decrease in the ROW of the treated groups of rats when compared to the control group. In biochemical analysis, there was a significant increase in aspartate aminotransferase, creatinine, urea, and uric acid. Hematological parameters showed a significant decrease in leukocytes, eosinophil, basophil, lymphocytes, monocytes, and hematocrit. Histopathological evaluation which revealed major histology changes of liver sections of rats treated with low and medium doses had lymphocyte and plasma cell inflammatory infiltrates; whereas, the liver sections of rats treated with high, lethal, and traditional doses exhibited lymphocyte, plasma cell, and eosinophilic inflammatory infiltrates. Conclusion: The study finds that the plants' mixture marketed by herbalists for the treatment of kidney diseases is toxic to body organs such as the liver and the hematopoietic system.

Bioactive secondary metabolites from the locally isolated terrestrial fungus, Penicillium sp. SAM16-EGY Mosad A Ghareeb, Manal M Hamed, Amal M Saad, Mohamed S Abdel-Aziz, Ahmed A Hamed, Laila A Refahy Pharmacognosy Research 2019 11(2):162-170 Background: Penicillium is a diverse genus occurring worldwide; its species are of major importance in the natural environment as decomposer of organic materials as well as food and drug production. Objective: Chromatographic isolation and identification of its bioactive secondary metabolites and their evaluation as antimicrobial agents. Materials and Methods: Disc agar plate method has been recognized to assess the antimicrobial activities. The antioxidant activity was determined using phosphomolybdenum method. The fungus strain SAM16-EGY was isolated from soil and was molecularly identified as Penicillium sp. SAM16-EGY using 18S ribosomal ribonucleic acid technique (acc. no., KP125952). Results: Seven compounds namely 3-O-docosyl-4-benzoyloxy methyl-3-oxobicyclo (4.1.0) heptane-1,5,6,7 tetrol (3-O-docosyl-3-debenzol rotepoxide) (1), (4bE, 6Z, 8E, 9aS, 10S)-1,4-dihydroxy-9a, 10-dihydro-10,12-epoxy-5-methylbenzo[a] azulen-12-one (2), 7α,9β,15β-triacetoxy-3-β-hydroxy jatropha-5E, 11E-diene (3), sesquiterpene I diol dihexoside malonate ester (4), piperogalone (5), (5R, 8Z, 11Z)-5 β-(6'-O-malonyl-β-glucopyranozyloxy-6-hydroxy tetradeca-8, 11-dienoic acid (6), and n-trcosanyl-n-octaced-9-enoate (7) were isolated and identified from this fungus. Their structures were determined on the basis of proton nuclear magnetic resonance and carbon-13 nuclear magnetic resonance spectroscopy. Compounds 1, 2, 4, and 5 exhibited antimicrobial activities against Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans only, whereas compound 3 exerted higher antimicrobial activity against S. aureus (9 mm), P. aeruginosa (9 mm), C. albicans (11 mm), and Aspergillus niger (13 mm) as compared to the other compounds. In the phosphomolybdenum assay, compound 5 showed high total antioxidant capacity value of 608.59 mg ascorbic acid equivalent/g compound, followed by compound 2 (443.66 mg) and compound 1 (332.16 mg). Conclusion: The isolated compounds showed promising antimicrobial and antioxidant activities.