International Journal of

Toxicological and Pharmacological Research

e-ISSN: 0975 5160

p-ISSN: 2820-2651

Peer Review Journal

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1 Antibacterial Activity and Toxicological Analysis In silico of 7-Hydroxycitronellal
Araújo M I F, Freitas F O R, Morais A M B, Brusten V P, Nogueira R B Sá S, Nogueira T B Sá S, Sousa M N A, Almeida Filho G G, Lima E O, Medeiros C I S
The pharmacological selectivity imposed by the micro-organisms enabled the emergence of new perspectives in the research field of natural products, in which the monoterpenic phytoconstituents, especially the 7-hydroxycitronellal (7-OH) present antimicrobial activity. Therefore, this study assessed the in silico antimicrobial and toxicological activity of the monoterpene 7-OH. The product was solubilized in tween 80 and dimethylsulfoxide (DMSO). Later it was diluted in sterile distilled water up to the concentration of 2048µg/mL. The minimum inhibitory concentration (MIC) of the product was determined by microdilution in doubly concentrated brain heart infusion (BHI) medium. The minimum bactericidal concentration (MBC) was determined by the depletion technique in nutrient agar (NA) with aliquots of 10µL of the MIC, MIC × 2 and MIC × 4. The MIC and the MBC of the 7-OH was 64µg/mL for S. aureus (ATCC 6538). However, for S. epidermides (LPM 35), S. aureus (LPM 45) and S. aureus (LPM 55), were 512 and 1024µg/mL respectively. In the in silico toxicological analysis the absence of mutagenic, tumorigenic effects and damage to the reproductive system were observed. Therefore, the 7-OH is bactericidal for the strains used in this study and presents a theoretical low oral toxicity.

2.Dicentra scandens (D.Don) Walp. – A Potential Source of Antimicrobial Agent
Koirala Pramila, Singh Bimala
Dicentra scandens (D.Don) Walp. locally called as ‘Jogi Lahara’ belongs to the Family Fumariaceae and is used in traditional medicine in Sikkim, a North Eastern state of India. The present study investigated the antimicrobial properties of aqueous and methanol extracts of Dicentra scandens (D.Don) Walp. against some test Gram negative and Gram positive bacteria. Using agar well diffusion method, aqueous and methanol extracts of roots of Dicentra scandens were tested against Escherichia coli (MTCC 1089), Klebsiella pneumoniae (MTCC 3384), Pseudomonas aeruginosa (MTCC 1034), Proteus vulgaris (MTCC 742), Salmonella typhi (MTCC 733), Shigella flexneri (MTCC 1457), Vibrio cholerae O139(MTCC 3906), Bacillus cereus (MTCC 6840) and Staphylococcus aureus (MTCC 7443). Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were also evaluated. Promising antimicrobial activity was exhibited by methanol extract of Dicentra scandens. The methanol extract was further characterized by Gas Chromatography-Mass Spectrometry (GC-MS) analysis. The GC-MS analysis revealed 29 compounds and the major compounds detected were Protopine (53.78%) and Corydine (18.20%). Thus, the alkaloids are predominant phytoconstituents of the extract and could be attributed to its antimicrobial activity. The results of the present study indicate that, Dicentra scandens can be a source of potential antimicrobial agent and can be explored further for its therapeutic use.

3. In Vitro Study on Quercetin Metabolism and Transport Rate in Presence of Benzo(a)pyrene
Naccari C, Kroon P A
Quercetin is a flavonoid able to modify the metabolism of carcinogen benzo(a)pyrene in human intestinal cells but it is not known if its metabolism is affected by the presence of B(a)P. In this study the phase-2 metabolism and transport rate of quercetin has been studied using CaCo-2/TC7 cell line, model of human enterocytes, co-treated with B(a)P.  Quercetin and its main metabolites, sulfated and glucuronidated, were quantified at different times (from T0 to T 24h), using HPLC with diode array and MS detection. This study showed that quercetin metabolism in CaCo-2/TC7 cells co-treated with B(a)P presents some differences respect to control in the formation of sulphates coniugates, more evident in the last time of analysis. The transport study, instead, confirmed a stronger influence of BaP with an increased rate of glucuronidation and sulfation of quercetin in both cellular side. The effects of B(a)P observed on phase-2 metabolism and transport rate of quercetin, by in vitro model of human enterocytes, put in evidence its possible influence, consequently, also on biological activities of this flavonoid.

4.In Vivo Antimalarial Activity of Adansonia digitata Stem Bark Extract and Some Fractions
Adeoye Akinwunmi O, Bewaji Clement O, Ademowo George O
Background: Malaria is one of the most common major health problems responsible for the death of millions of children, pregnant women and adults. Antimalarial drug resistance has emerged as one of the greatest challenges facing malaria control today. Plant resources that either treat or prevent parasite invasion desirable in developing countries are potential targets for research and development of alternative malaria drugs. Objective: This study investigated the suppressive and prophylactic potentials of extracts and some fractions of Adansonia digitata stem bark in Plasmodium berghei infected mice. Methodology: The albino mice were administered with two different doses (200mg/kg body weight and 400mg/kg body weight) of aqueous extract (AQ), methanolic extract (ME), chloroform fraction (CF) and ethylacetate fraction (EF) of Adansonia digitata stem bark for five consecutive days. 5mg/kg body weight dose per day of artemether-lumefantrine and 5mg/kg body weight dose per day of chloroquine was used as positive control while the negative control mice received only the vehicle (5% v/v tween 80). In the prophylactic groups, the mice were pretreated daily for five days before they were challenged with inoculums of 1 x 107 chloroquine-sensitive P. berghei infected erythrocyte intraperitoneally. Results: The results showed a dose dependent chemosupression in the fractions and the extract treated groups. The 400mg/kg body weight was more effective with respect to the parasite clearance than the 200mg/kg body weight in all the groups. Both the 200mg/kg and 400mg/kg body weight dose of ethylacetate fraction (EF) exhibited the highest chemosupression. The chemosupression caused by Artemether-lumefantrine (AL) and Chloroquine (CQ) treated groups were significantly (P< 0.05) higher than the fractions and extract treated groups. The percentage parasitemia also decreased in this manner. There was a mutual delay in parasitemia with EF and ME. The packed cell volume (PCV) increased significantly (P< 0.05) in the AL and CQ, and 400mg/kg body weight dose EF and ME respectively and increased for the other fraction and extract used at 400mg/kg body weight dose compared with the control. Conclusion: This study showed that EF of Adansonia digitata stem bark has potent antimalarial property which could be of future importance in malaria management.

5.Standardisation and Validation of Acute and Chronic Administration Ketamine at Different Doses to Produce Psychosis Like Behavioural Changes in Mice
Monu Yadav, Milind Parle, Mamta Sachdeva, Sameer Dhingra
Psychosis is a complex mental illness, characterised by positive, negative and cognitive symptoms. NMDA receptor antagonists have been established to induce behavioural as well as biochemical changes in rodents similar to psychotic patients. The aim of the present study was to investigate the effective dose and treatment period of ketamine to induce some behavioural changes. The results suggest that acute treatment of ketamine (50 and 100 mg/kg, i.p.) induced hyperlocomotor activity and reduced step down latency time in passive avoidance test, whereas in effective in forced swim test. Further, with the chronic administration of ketamine (50 and 100 mg/kg, i.p.) effective to produced hyperlocomotor activity, reduced the step down latency time in passive avoidance test and enhanced the immobility duration in forced swim test. Moreover, these behavioural changes persisted for 7 days after the treatment period. Thus, our findings suggest that the chronic administration of Ketamine (50 and 100 mg/kg, i.p.) potential to produce behavioural changes, would serve as an effective tool to screen antipsychotic drugs.

6. Anxiolytic Effect and Acute Toxicity of Two Novel Combinations of Plant Extracts – Antistress I and Antistress II on Rats
Kandilarov I, Zlatanova H, Delev D, Kostadinova I, Katsarova M, Dimitrova S, Lukanov L, Sadakov F
Anxiety is a significant social problem that could deteriorate the quality of life of individuals. It disturbs the normal circadian rhythms, leading to an increase of cardio-vascular diseases and distortions in the functions of the immune system. Antistress I and Antistress II are herbal combinations containing in different proportions dry extracts obtained from Serratula coronata, Hypericum perforatum, Valeriana officinalis, Crataegus monogyna and Melissa officinalis. The present study evaluates the acute toxicity and the anxiolytic effect of these combinations and the individual extracts which they contain after their oral administration to male Wistar rats. Doses of 5g/kg b.w. and 10g/kg b.w. are used for the evaluation of acute toxicity. Assessment of the anxiolytic effect is carried out at three doses – 100, 250 and 500 mg/kg b.w. by using the test – Elevated plus maze. The results about the acute toxicity show a survival rate of 100% for all extracts at a dose up to10g/kg b.w. The evaluation of the anxiolytic effect on an acute stress model in rats demonstrates that both combinations Antistress I and Antistress II possess anxiolytic properties which are significant only at the highest dose.  The results also give us the reason to conclude that the effect of Antistress II on anxiety is better compared to Antistress I.

7. Screening of Antianxiety Activity of Extracts and Fractions of Verbena officinalis Aerial Parts
Pawan Kumar, Reecha Madaan, Shabir Sidhu
Verbena officinalis L. (Vervain; family-Verbenaceae) has been traditionally used in the treatment of various ailments especially in mental disorders, but no systematic work has been carried out to validate traditional claims of plant. Therefore, it was planned to screen antianxiety activity of extracts and fractions of V. officinalis aerial parts. Crude extracts of V. officinalis aerial parts were prepared by extracting the plant material successively with n-hexane, chloroform, methanol and water. The antianxiety activity of crude extracts was performed at the doses of 200 or 400 mg/kg, p.o., upon acute administration in mice using elevated plus maze model. The bioactive extract was partitioned using ethyl acetate, and ethyl acetate fraction (50 or 100 mg/kg, p.o.) and remaining bioactive extract (150 or 300 mg/kg, p.o.) were also screened for antianxiety activity. The statistical significance was checked by comparing with standard drug and control using one way analysis of variance (ANOVA), which was followed by post hoc analysis – Student-Newman-Keul’s test. The methanol extract and ethyl acetate fraction exhibited significant antianxiety activity at the dose of 400 mg/kg and 100 mg/kg, respectively, with respect to control and statistically equivalent to the standard drug (Diazepam, 2 mg/kg, p.o.). The bioactive ethyl acetate fraction is rich in phenolic compounds and flavonoids. Finally, it is concluded that these are major anxiolytic constituents of V. officinalis.

8. Estimation of Proximate, Micronutrients and Glycaemic Index of Talas Safira (Colocasia esculenta var. Antiquorum Schott) Growing in Bantaeng-Indonesia
Lukman M, Adrianus Suparman, Wahyu Hendrarti, Abdul Halim Umar, Yuri P Utami, Marwati, Aisyah Fatmawaty
Bantaeng people cultivate Talas safira (Colocasia esculenta var. Antiquorum Schott) and commonly eat it as a staple food in their daily as carbohydrate sources. The effect of carbohydrate to glucose blood level can be refer to as glycaemic index (GI) and glycaemic load (GL). A diet with a high GI and GL are associated with an elevated risk of type 2 diabetes and obesity. We investigated proximate, micronutrient, heavy metal, GL and GI of Talas safira which cultivated in Bantaeng. The result revealed marked that Talas safira contain carbohydrate; essential micronutrients and rich of energy (380.55 kJ/100 g). High content carbohydrate of Talas safira refer to high level of GL (high) and GL (high). This results indicated that Talas safira needs attention to people with metabolic glucose disorder.

9. Phytochemical and Pharmacological Activity of Myristica fragrans  Houtt (Myristicaceae)
Md Arifur Rahman Chowdhury, Manirujjaman, Md Mazedul Haq
Objective: Myristica fragrans Houtt commonly used as traditional medicine for alleviating of various disorders. The purpose of our study was to map out the in vitro antioxidant property and in vivo anti-hyperglycemic and analgesic effect of the methanolic extract of Myristica fragrans Houtt. (Seed and mace) (Myristicaceae) on Swiss albino mice. Methods: The processed powder of Myristica fragrans Houtt (seed and mace) were subjected to methanolic extraction by soxhlet filtration methods, and the desiccated extract was used for screening of antioxidant by DPPH free radical scavenging assessment as well as total phenolic content by using folin-ciocalteu reagent.Anti-hyperglycemic effect and analgesic action  tested through alloxan induced antidiabetics test and acetic acid-tempted writhing test on mice. Results: In DPPH free radical scavenging assessment, free radicals neutralization expressed as % of inhibition 49.69±0.06% also by IC50  values as 68.43 µg/ml surmise middle level of antioxidant property. The total phenolic content expressed as 186.25 mg/g equivalent of gallic acid indicates, active phenolic content. Oral administration of 200 and 400 mg/kg of extract dose and reference drug vildagliptin (50 mg/kg) for the duration of the 4-day study period, and initiated % of inhibition the blood glucose level measured as 22.48%, 44.78% and 62.02% regard as the significant anti-hyperglycemic properties. The analgesic activity was investigated by using the acetic acid-induced writhing test in mice, at the dose of 200 mg/kg body and 400mg/kg weight, and resulting 50.4% and 68.10% correspondingly, which was considerably significant with a standard drug. Conclusion: The present study suggests that methanolic extract of seed and mace of Myristica fragrans Houtt can manage moderate oxidative stress as well as perform the painkilling action. Besides, prolong medication may enhance the new dimension of anti-hyperglycemic activity.

10. Antimicrobial Properties of Proton Pump Inhibitors
Patil T R, Patil S, Patil A, Patil S T
Proton pump inhibitors [PPI] are acid activated pro drugs which convert in to sulfenic acid and then in to tetracycline sulfenamide in the acidic pH of parietal cell canaliculi. They block H+K+ATPase proton pumps and reduce gastric acid secretion. They are used to treat acid peptic disorders and NSAID induced gastric mucosal injury. Activity of PPI against Helicobactor pylori [H.pylori] is proved undisputably. Lansoprazole is the most effective PPI against H.pylori due to its unique chemical structure.PPI inhibit urease activity of H. pylori. They affect respiration and energy metabolism of these organisms as result of decreased ATP synthesis. Structural similarity of benzimidazole PPI with imidazole like metronidazole and tinidazole may contribute for their antibacterial property. Omeprazole and lansoprazole have been found to have anti fungal activity by inhibition of fungal H+K+ATPase-vacuolar ATPase which are essential for fungal survival and to carry out essential physiological functions, the inhibition of which leads to fungicidal action. Recently anti tubercular action of lansoprazole was highlighted which is attributed to its intra mycobacterial sulfoxide reduction to lansoprazole sulfide. This acts on mycobacterial cytochrome bc1 complex and inhibits ATP synthesis and compromises energy metabolism threatening its survival. Cytochrome bc1 of plasmodium also forms a drug target for lansoprazole. Thus, lansoprazole can emerge as a potential drug to treat MDR tuberculosis and malaria. Antiviral action of lansoprazole was noted against rhinovirus. Gram positive and negative organisms other than H.pylori were found to be inhibited by omeprazole in vitro. But this is not supported by in vivo studies.

11. Role of Vitamin D3 in Prevention of Paracetamol Induced Acute Hepatotoxicity in Male Rabbits
Alsaady Malath Azeez
Background: Liver is major organ of metabolism, which required to be protected from drugs and their metabolites harmful effect by using substances like vitamins and herbs. Objective: To evaluate vit. D3 protective effect in paracetamol induced hepatotoxicity of male rabbits. Material and method:18 male local rabbits divided to three groups. First group receive paracetamol in three spaced doses for 24 hours to induce acute hepatoxicity while the second group receive vit.D3 as single intramuscular injection 24 hours prior induction of acute hepatoxicity and third group was control without treatment. After 24 hours from paracetamol, induce acute hepatotoxicity blood samples collected to measure alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and albumin. Results: There were significant elevation (p≤0.05) in serum liver enzymes of treatment groups in relation to control and non-significant decrease in alanine aminotransferase (ALT), aspartate aminotransferase (AST) of vit.D3 treated group. Conclusion: As conclusion, a model of acute paracetamol induced hepatotoxicity successfully established with no mortality.  Although, non-significant vit.D3 possess protective effect in relation to liver function test.

12. Evaluation of Aphrodisiac Activity of Phaseolus semierectus (L.) Seeds
Neerugatti Dorababu, Battu Ganga Rao, Devarakonda Ramadevi
The study was aimed at investigate the effect of methanolic extract of Phaseolus semierectus (L.) (Leguminosae) on general  mating  behaviour, libido, and adverse effects on sexually normal male albino rats.  Methanolic extract was administered orally at the dose of 100, 200, and 400 mg / kg, to different groups of male rats (n = 8) once a day for 14 days. All the doses resulted in significant increase in mount frequency, intromission frequency and anogenital sniffing when compared to normal. The methanolic extract of Phaseolus semierectus (L.) seeds at higher concentration (400 mg/kg body weight) showed significant aphrodisiac activity on male Wister albino rats as evidenced by an increase in number of mounts and mating performance. Thus, in experimental rats, the results of the present study suggest that the methanolic extracts of Phaseolus semierectus (L.) exert significant aphrodisiac activity.

13. A Review: Uses of Gas Chromatography-Mass Spectrometry (GC-MS) Technique for Analysis of Bioactive Natural Compounds of Some Plants
Abeer Fauzi Al-Rubaye, Imad Hadi Hameed, Mohanad Jawad Kadhim
Chromatography is the term used to describe a separation technique in which a mobile phase carrying a mixture is caused to move in contact with a selectively absorbent stationary phase. It also plays a fundamental role as an analytical technique for quality control and standardization of phyto therapeuticals. Gas Chromatography is used in the separation and analysis of multi component mixtures such as essential oils, hydrocarbons and solvents. Various temperature programs can be used to make the readings more meaningful; for example to differentiate between substances that behave similarly during the GC process. Intrinsically, with the use of the flame ionization detector and the electron capture detector (which have very high sensitivities) gas chromatography can quantitatively determine materials present at very low concentrations. Plants are a rich source of secondary metabolites with interesting biological activities. In general, these secondary metabolites are an important source with a variety of structural arrangements and properties. Gas chromatography – specifically gas-liquid chromatography – involves a sample being vapourised and injected onto the head of the chromatographic column. The sample is transported through the column by the flow of inert, gaseous mobile phase. The column itself contains a liquid stationary phase which is adsorbed onto the surface of an inert solid. The principle of gas chromatography is adsorption and partition. Within the family of chromatography- based methods gas chromatography (GC) is one of the most widely used techniques. GC-MS has become a highly recommended tool for monitoring and tracking organic pollutants in the environment. GC-MS is exclusively used for the analysis of esters, fatty acids, alcohols, aldehydes, terpenes etc. It is the key tool used in sports anti-doping laboratories to test athlete’s urine samples for prohibited performanceenhancing drugs like anabolic steroids. Several GC-MS have left earth for the astro chemistry studies.  As a unique and powerful technology the GC-MS provides a rare opportunity to perform the analysis of new compounds for characterization and identification of synthesized or derivatized compound.

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